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Obstetrician & Gynecologist (OB/GYN)
7 years of experience
Accepting new patients

Credentials

Education ?

Medical School Score Rankings
The University of Texas at Houston (2005)
  •  
Top 50%

Awards & Distinctions ?

Associations
American Board of Obstetrics and Gynecology

Affiliations ?

Dr. Phillips is affiliated with 1 hospitals.

Hospital Affiliations

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Rankings

  • Medical Center Of Arlington
    3301 Matlock Rd, Arlington, TX 76015
    •  
    Top 50%
  • Publications & Research

    Dr. Phillips has contributed to 206 publications.
    Title Spatio-temporal Theory of Lasing Action in Optically-pumped Rotationally Excited Molecular Gases.
    Date October 2011
    Journal Optics Express
    Excerpt

    We investigate laser emission from optically-pumped rotationally excited molecular gases confined in a metallic cavity. To this end, we have developed a theoretical framework able to accurately describe, both in the spatial and temporal domains, the molecular collisional and diffusion processes characterizing the operation of this class of lasers. The effect on the main lasing features of the spatial variation of the electric field intensity and the ohmic losses associated to each cavity mode are also included in our analysis. Our simulations show that, for the exemplary case of methyl fluoride gas confined in a cylindrical copper cavity, the region of maximum population inversion is located near the cavity walls. Based on this fact, our calculations show that the lowest lasing threshold intensity corresponds to the cavity mode that, while maximizing the spatial overlap between the corresponding population inversion and electric-field intensity distributions, simultaneously minimizes the absorption losses occurring at the cavity walls. The dependence of the lasing threshold intensity on both the gas pressure and the cavity radius is also analyzed and compared with experiment. We find that as the cavity size is varied, the interplay between the overall gain of the system and the corresponding ohmic losses allows for the existence of an optimal cavity radius which minimizes the intensity threshold for a large range of gas pressures. The theoretical analysis presented in this work expands the current understanding of lasing action in optically-pumped far-infrared lasers and, thus, could contribute to the development of a new class of compact far-infrared and terahertz sources able to operate efficiently at room temperature.

    Title Pattern of Activin A and Follistatin Release in a Sheep Model of Cardiopulmonary Bypass.
    Date July 2011
    Journal Cytokine
    Excerpt

    Activin A, a member of transforming growth factor-β superfamily, has been established as a critical cytokine released early in endotoxemia and other inflammatory syndromes. The release of activin A and its binding protein, follistatin during cardiopulmonary bypass (CPB) has not been previously reported. Our study aimed to define the pattern of activin A and follistatin release in a sheep CPB model.

    Title The Regulation and Functions of Activin and Follistatin in Inflammation and Immunity.
    Date June 2011
    Journal Vitamins and Hormones
    Excerpt

    The activins are members of the transforming growth factor β superfamily with broad and complex effects on cell growth and differentiation. Activin A has long been known to be a critical regulator of inflammation and immunity, and similar roles are now emerging for activin B, with which it shares 65% sequence homology. These molecules and their binding protein, follistatin, are widely expressed, and their production is increased in many acute and chronic inflammatory conditions. Synthesis and release of the activins are stimulated by inflammatory cytokines, Toll-like receptor ligands, and oxidative stress. The activins interact with heterodimeric serine/threonine kinase receptor complexes to activate SMAD transcription factors and the MAP kinase signaling pathways, which mediate inflammation, stress, and immunity. Follistatin binds to the activins with high affinity, thereby obstructing the activin receptor binding site, and targets them to cell surface proteoglycans and lysosomal degradation. Studies on transgenic mice and those with gene knockouts, together with blocking studies using exogenous follistatin, have established that activin A plays critical roles in the onset of cachexia, acute and chronic inflammatory responses such as septicemia, colitis and asthma, and fibrosis. However, activin A also directs the development of monocyte/macrophages, myeloid dendritic cells, and T cell subsets to promote type 2 and regulatory immune responses. The ability of both endogenous and exogenous follistatin to block the proinflammatory and profibrotic actions of activin A has led to interest in this binding protein as a potential therapeutic for limiting the severity of disease and to improve subsequent damage associated with inflammation and fibrosis. However, the ability of activin A to sculpt the subsequent immune response as well means that the full range of effects that might arise from blocking activin bioactivity will need to be considered in any therapeutic applications.

    Title Cortical Evoked Responses Associated with Arousal from Sleep.
    Date May 2011
    Journal Sleep
    Excerpt

    To determine if low-level intermittent auditory stimuli have the potential to disrupt sleep during 24-h recordings, we assessed arousal occurrence to varying stimulus intensities. Additionally, if stimulus-generated evoked response potential (ERP) components provide a metric of underlying cortical state, then a particular ERP structure may precede an arousal.

    Title Predictive Factors of Hearing Preservation After Surgical Resection of Small Vestibular Schwannomas.
    Date March 2011
    Journal Otology & Neurotology : Official Publication of the American Otological Society, American Neurotology Society [and] European Academy of Otology and Neurotology
    Excerpt

    To identify factors predictive of hearing preservation in patients undergoing resection of small vestibular schwannoma.

    Title Thermal Motion of Tert-butyl Groups Iii. Tert-butyl Substituents in Aromatic Hydrocarbons, the View from the Bottom of the Well.
    Date December 2010
    Journal Acta Crystallographica. Section B, Structural Science
    Excerpt

    The rigidity of the tert-butyl group (TBG) as a substituent in aromatic hydrocarbons is investigated, with a modified Hirshfeld test of anisotropic displacement parameters (ADPs) as a primary criterion. Four new structures are analyzed, along with low-temperature studies of a previously published crowded supermesityl dimer; three of the five structures meet the primary test. Most of the TBGs meet the Hirshfeld test at 100 K, and the ADPs are improved by omitting low-order data in the final refinement. The three most precise structures yield a wide variation in libration amplitudes (and in estimated rotation barriers) for 13 unique TBGs. A similar range of values is found in analyses of structures in the Cambridge Crystallographic Database. The libration amplitudes are calculated with the program THMA14C, with each TBG as an attached rigid group (ARG). Packing analysis suggests that large ADPs, especially for some individual TBG methyl groups, correspond to voids in the crystal. Published barriers to TBG reorientation, determined by solid-state NMR spin-lattice relaxation methods, for six related crystalline compounds are compared with barriers calculated from their crystal structure data.

    Title Interleukin-13 Regulates Secretion of the Tumor Growth Factor-{beta} Superfamily Cytokine Activin A in Allergic Airway Inflammation.
    Date June 2010
    Journal American Journal of Respiratory Cell and Molecular Biology
    Excerpt

    Activin A is a member of the TGF-beta superfamily and plays a role in allergic inflammation and asthma pathogenesis. Recent evidence suggests that activin A regulates proinflammatory cytokine production and is regulated by inflammatory mediators. In a murine model of acute allergic airway inflammation, we observed previously that increased activin A concentrations in bronchoalveolar lavage (BAL) fluid coincide with Th2 cytokine production in lung-draining lymph nodes and pronounced mucus metaplasia in bronchial epithelium. We therefore hypothesized that IL-13, the key cytokine for mucus production, regulates activin A secretion into BAL fluid in experimental asthma. IL-13 increased BAL fluid activin A concentrations in naive mice and dose dependently induced activin A secretion from cultured human airway epithelium. A key role for IL-13 in the secretion of activin A into the BAL fluid during allergic airway inflammation was confirmed in IL-13-deficient mice. Eosinophils were not involved in this response because there was no difference in BAL fluid activin A concentrations between wild-type and eosinophil-deficient mice. Our data highlight an important role for IL-13 in the regulation of activin A intraepithelially and in BAL fluid in naive mice and during allergic airway inflammation. Given the immunomodulatory and fibrogenic effects of activin A, our findings suggest an important role for IL-13 regulation of activin A in asthma pathogenesis.

    Title Diagnosis and Outcomes of Middle Cranial Fossa Repair for Patients with Superior Semicircular Canal Dehiscence Syndrome.
    Date April 2010
    Journal Journal of Clinical Neuroscience : Official Journal of the Neurosurgical Society of Australasia
    Excerpt

    The aim of this study was to retrospectively review the clinical presentation, diagnostic features, in particular cervical vestibular evoked myogenic potentials (cVEMPs), and the outcomes of surgical repair for superior semicircular canal dehiscence syndrome (SSCDS). SSCDS is a well-described syndrome of auditory and vestibular symptoms due to a bony dehiscence of the superior semicircular canal in the middle cranial fossa. A series of six procedures on five patients with SSCDS who underwent surgical repair via a middle fossa craniotomy were retrospectively reviewed. Preoperative and postoperative audiometric and vestibular symptoms as well as investigation findings were reviewed. Auditory and vestibular symptoms improved and hearing was preserved in all patients. The low frequency pseudo-conductive loss was corrected in four out of five patients, and the lowered preoperative cVEMP thresholds normalised following successful middle cranial fossa repair. In this series, middle fossa repair of SSCD was safe and effective with excellent sensorineural hearing preservation.

    Title Acute and Chronic Effects of Endotoxin on Cerebral Circulation in Lambs.
    Date March 2010
    Journal American Journal of Physiology. Regulatory, Integrative and Comparative Physiology
    Excerpt

    The impact of endotoxemia on cerebral endothelium and cerebral blood flow (CBF) regulation was studied in conscious newborn lambs. Bacterial endotoxin [LPS, 2 microg/kg iv] was infused on 3 consecutive days. Cerebrovascular function was assessed by monitoring CBF and cerebral vascular resistance (CVR) over 12 h each day and by the endothelium-dependent vasodilator bradykinin (BK) (n = 10). Inflammatory responses were assessed by plasma tumor necrosis factor-alpha (TNF-alpha, n = 5). Acutely, LPS disrupted the cerebral circulation within 1 h, with peak cerebral vasoconstriction at 3 h (CBF -28 and CVR +118%, P < 0.05) followed by recovery to baseline by 12 h. TNF-alpha and body temperature peaked approximately 1 h post-LPS. BK-induced vasodilatation (CVR -20%, P < 0.05) declined with each LPS infusion, was abolished after 3 days, and remained absent for at least the subsequent 5 days. Histological evidence of brain injury was found in four of five LPS-treated newborns. We conclude that endotoxin impairs cerebral perfusion in newborn lambs via two mechanisms: 1) acute vasoconstriction (over several hours); and 2) persistent endothelial dysfunction (over several days). Endotoxin-induced circulatory impairments may place the newborn brain at prolonged risk of CBF dysregulation and injury as a legacy of endotoxin exposure.

    Title Activin and Related Proteins in Inflammation: Not Just Interested Bystanders.
    Date June 2009
    Journal Cytokine & Growth Factor Reviews
    Excerpt

    Activin A, a member of the transforming growth factor-beta superfamily, is released rapidly into the circulation during inflammation. This review examines the evidence that activin is a critical mediator of inflammation and immunity. Activin modulates several aspects of the inflammatory response, including release of pro-inflammatory cytokines, nitric oxide production and immune cell activity. Crucially, inhibiting activin with follistatin, a high affinity binding protein, alters the pattern of cytokines released and improves survival in a mouse model of endotoxic shock. Serum and tissue concentrations of activin are elevated in a wide range of pathological conditions. The utility of activin as a diagnostic marker of clinical inflammation and the use of follistatin to block activin actions therapeutically are also discussed.

    Title A New Role for Activin in Endometrial Repair After Menses.
    Date April 2009
    Journal Endocrinology
    Excerpt

    Abnormal uterine bleeding can severely affect the quality of life for women. After menstruation, the endometrium must adequately repair to limit and stop bleeding. Abnormal uterine bleeding may result from incorrect or inadequate endometrial repair after menstruation. Previous studies have shown an important contribution of activin to skin wound healing, with severely delayed wound repair observed in animals transgenically induced to overexpress activin's natural inhibitor, follistatin. Activin subunits have also been identified within human endometrium; however, their role in endometrial repair is unknown. We assessed the contribution of activin to endometrial repair after menses using a human in vitro cell wounding method and our well-characterized mouse model of endometrial breakdown and repair applied to mice overexpressing follistatin. Endometrial repair after menses is initiated with reepithelialization of the uterine surface. To mimic this repair, we utilized a human endometrial epithelial cell line (ECC-1) and demonstrated significant stimulation of wound closure after activin A administration, and attenuation of this response by addition of follistatin. Immunolocalization of activin subunits, betaA and betaB, in control endometrium from the mouse model demonstrated specific epithelial and stromal localization and some leukocyte staining (betaA) around sites of endometrial repair, suggestive of a role for activin in this process. Follistatin-overexpressing animals had significantly higher circulating follistatin levels than wild-type littermates. There was a significant delay in endometrial repair after breakdown in follistatin transgenic animals compared with control animals. This study demonstrates for the first time a functional role for activin in endometrial repair after menses.

    Title Endotoxin Has Acute and Chronic Effects on the Cerebral Circulation of Fetal Sheep.
    Date April 2009
    Journal American Journal of Physiology. Regulatory, Integrative and Comparative Physiology
    Excerpt

    We studied the impact of endotoxemia on cerebral blood flow (CBF), cerebral vascular resistance (CVR), and cerebral oxygen transport (O(2) transport) in fetal sheep. We hypothesized that endotoxemia impairs CBF regulation and O(2) transport, exposing the brain to hypoxic-ischemic injury. Responses to lipopolysaccharide (LPS; 1 microg/kg iv on 3 consecutive days, n = 9) or normal saline (n = 5) were studied. Of LPS-treated fetuses, five survived and four died; in surviving fetuses, transient cerebral vasoconstriction at 0.5 h (DeltaCVR approximately +50%) was followed by vasodilatation maximal at 5-6 h (DeltaCVR approximately -50%) when CBF had increased (approximately +60%) despite reduced ABP (approximately -20%). Decreased CVR and increased CBF persisted 24 h post-LPS and the two subsequent LPS infusions. Cerebral O(2) transport was sustained, although arterial O(2) saturation was reduced (P < 0.05). Histological evidence of neuronal injury was found in all surviving LPS-treated fetuses; one experienced grade IV intracranial hemorrhage. Bradykinin-induced cerebral vasodilatation (DeltaCVR approximately -20%, P < 0.05) was abolished after LPS. Fetuses that died post-LPS (n = 4) differed from survivors in three respects: CVR did not fall, CBF did not rise, and O(2) transport fell progressively. In conclusion, endotoxin disrupts the cerebral circulation in two phases: 1) acute vasoconstriction (1 h) and 2) prolonged vasodilatation despite impaired endothelial dilatation (24 h). In surviving fetuses, LPS causes brain injury despite cerebral O(2) transport being maintained by elevated cerebral perfusion; thus sustained O(2) transport does not prevent brain injury in endotoxemia. In contrast, cerebral hypoperfusion and reduced O(2) transport occur in fetuses destined to die, emphasizing the importance of sustaining O(2) transport for survival.

    Title Imidazopyridines As Vla-4 Integrin Antagonists.
    Date November 2008
    Journal Bioorganic & Medicinal Chemistry Letters
    Excerpt

    We describe a novel series of imidazopyridine substituted phenylalanines which are potent VLA-4 antagonists. A wide variety of substituents are tolerated as replacements for the pendant 3-pyridyl ring. A clear structure-activity relationship was identified around the substitution of the 3-amino-cyclobut-2-enone portion of the molecule.

    Title Securing the Oral Tradition: Reflective Positioning and Professional Conversations in Midwifery Education.
    Date November 2008
    Journal Collegian (royal College of Nursing, Australia)
    Excerpt

    We postulate that positioning is a powerful tool in guiding and transforming student professional learning and practice development. In our experiences with students enrolled in he Graduate Diploma of Midwifery, we determined that positioning elaborates individual scholarship and identity formation of the learner midwife in practice settings. Positioning Theory, developed by Harré and other authorities, is a psycho-sociological 'ontology' or concept of how individuals metaphorically position or locate themselves, and others, within institutions and societies. Three key components of positioning theory include 'position', 'speech act' and 'storylines', developing from the everyday social interactions of professional conversations. Reflective positioning can be applied as an analytical tool for the moment-to-moment exchanges inherent in practice related conversations, occurring between midwives and midwifery students. These moment-to-moment interactions of professional conversations can be used by students to complete or fill their learning gaps. Positioning therefore, provides a novel, contemporary theoretical framework to 'unpack' or understand the complexity of midwifery practice and yet is complementary with reflective practice. Excerpts are used to demonstrate reflective positioning applications by students. Midwives are encouraged by health services and by the University to provide student support through a 'preceptorship' program to supervise, work with and assess students for competence in midwifery practices. We claim that reflective positioning by students within professional conversations with their preceptor/midwives, are the construction sites for learning and where identity formation of each student as a future midwife is both shaped and transformed. Both academics and managers of health services need to embrace the value of workplace conversations, the sites of rich oral traditions of nursing and midwifery. Thus, in seeking claim to our rich oral traditions, all students will benefit from engagement in reflective positioning to promote their professional learning and practice development.

    Title Characterization of Flexible Ecog Electrode Arrays for Chronic Recording in Awake Rats.
    Date October 2008
    Journal Journal of Neuroscience Methods
    Excerpt

    We developed a 64-channel flexible polyimide ECoG electrode array and characterized its performance for long-term implantation, chronic cortical recording and high resolution mapping of surface-evoked potentials in awake rats. To achieve the longest possible recording periods, the flexibility of the electrode array, adhesion between the metals and carrier substrate, and biocompatibility were critical for maintaining the signal integrity. Experimental testing of thin film adhesion was applied to a gold-polyimide system in order to characterize relative interfacial fracture energies for several different adhesion layers, yielding an increase in overall device reliability. We tested several different adhesion techniques including the following: gold alone without an adhesion layer, titanium-tungsten, tantalum and chromium. We found titanium-tungsten to be a suitable adhesion layer considering the biocompatibility requirements as well as stability and delamination resistance. While chromium and tantalum produced stronger gold adhesion, concerns over biocompatibility of these materials require further testing. We implanted the polyimide ECoG electrode arrays through a slit made in the skull of rats and recorded cortical surface evoked responses. The arrays performed reliably over a period of at least 100 days and signals compared well with traditional screw electrodes, with better high frequency response characteristics. Since the ultimate goal of chronically implanted electrode arrays is for neural prosthetic devices that need to last many decades, other adhesion layers that would prove safe for implantation may be tested in the same way in order to improve the device reliability.

    Title Testosterone Secretion, Testicular Histology and the Cryopreservation of Cauda Epididymidal Spermatozoa in the Common Ringtail Possum (pseudocheirus Peregrinus).
    Date June 2008
    Journal Reproduction, Fertility, and Development
    Excerpt

    The present study reports novel aspects of the reproductive biology of the male common ringtail possum (Pseudocheirus peregrinus). Plasma testosterone was measured through a stimulation test using the gonadotrophin-releasing hormone agonist, buserelin. Following intra-muscular administration of buserelin, there was an increase (P<0.05) in testosterone concentration in the peripheral circulation 4 h later. Quantitative testicular histology of this species was described for the first time. Eight stages of the seminiferous epithelium cycle were identified in 10 possums and their relative frequency determined. Spermatozoa were recovered from the cauda epididymides of hemi-castrated possums and cryopreservation conducted in straws (6 degrees C min(-1)) using final glycerol concentrations ranging between 2 and 20% in Tris-citrate egg yolk extender (v/v). Frozen straws were thawed and post-thaw motility, rate of motility, the percentage of live-dead spermatozoa and the percentage of sperm with swollen decondensed nuclei recorded. Similar to other marsupial sperm, common ringtail possum cauda epididymidal spermatozoa required high levels of glycerol (10-16%) in order to maintain post-thaw viability.

    Title Developmental Programming: Deficits in Reproductive Hormone Dynamics and Ovulatory Outcomes in Prenatal, Testosterone-treated Sheep.
    Date June 2008
    Journal Biology of Reproduction
    Excerpt

    Prenatal testosterone excess leads to neuroendocrine, ovarian, and metabolic disruptions, culminating in reproductive phenotypes mimicking that of women with polycystic ovary syndrome (PCOS). The objective of this study was to determine the consequences of prenatal testosterone treatment on periovulatory hormonal dynamics and ovulatory outcomes. To generate prenatal testosterone-treated females, pregnant sheep were injected intramuscularly (days 30-90 of gestation, term=147 days) with 100 mg of testosterone-propionate in cottonseed oil semi-weekly. Female offspring born to untreated control females and prenatal testosterone-treated females were then studied during their first two breeding seasons. Sheep were given two injections of prostaglandin F2alpha 11 days apart, and blood samples were collected at 2-h intervals for 120 h, 10-min intervals for 8 h during the luteal phase (first breeding season only), and daily for an additional 15 days to characterize changes in reproductive hormonal dynamics. During the first breeding season, prenatal testosterone-treated females manifested disruptions in the timing and magnitude of primary gonadotropin surges, luteal defects, and reduced responsiveness to progesterone negative feedback. Disruptions in the periovulatory sequence of events during the second breeding season included: 1) delayed but increased preovulatory estradiol rise, 2) delayed and severely reduced primary gonadotropin surge in prenatal testosterone-treated females having an LH surge, 3) tendency for an amplified secondary FSH surge and a shift in the relative balance of FSH regulatory proteins, and 4) luteal responses that ranged from normal to anovulatory. These outcomes are likely to be of relevance to developmental origin of infertility disorders and suggest that differences in fetal exposure or fetal susceptibility to testosterone may account for the variability in reproductive phenotypes.

    Title A Functional Epitope of the Pneumococcal Surface Adhesin A Activates Nasopharyngeal Cells and Increases Bacterial Internalization.
    Date May 2008
    Journal Microbial Pathogenesis
    Excerpt

    Pneumococcal surface adhesin A (PsaA) is a putative pneumococcal (Pnc) adhesin known to bind to nasopharyngeal (NP) epithelial cells. This study evaluated the effect of peptides within a functional domain of PsaA on NP cells. Detroit 562 NP cells were treated with synthetic peptides derived from PsaA (P4, P6, and P7; 28, 12, and 16 amino acids, respectively). The P4 peptide also binds to NP cells. Analysis of P4-treated NP cells by transmission electron microscopy revealed major cytological changes. Of 9 cytokines analyzed, a 6-fold increase in FGFb secretion at 3 and 6h (11-fold at 12h) was found post-P4 treatment of NP cells. There was a simultaneous reduction in the secreted levels of IL-6, IL-8, and VEGF. We observed enhancement in the adherence of Pnc strains to P4-treated NP cells (2-38-fold increase). Enhancement in adherence (2-fold increase) to P4-treated NP cells was also recorded with other streptococcal species (Streptococcus mitis and Streptococcus pyogenes). Internalization experiments demonstrated that 45% of the adherent bacteria were actually internalized after pretreatment with P4 peptide as compared to controls. Peptide fragments of P4, P6 and P7 did not activate NP cells to the extent of P4 peptide. The P4-mediated enhancement of Pnc adherence was blocked (100%) by anti-P4 antibodies, confirming the specificity of the P4 sequence for NP cell activation. Our data suggests that this functional domain of PsaA contained within the P4 sequence binds and activates NP cells to facilitate Pnc invasion.

    Title Temporal Expression of Activin in Acute Burn Wounds--from Inflammatory Cells to Fibroblasts.
    Date May 2008
    Journal Burns : Journal of the International Society for Burn Injuries
    Excerpt

    Activin A is a member of the transforming growth factor-beta (TGF-beta) family of cytokines and growth factors and upregulation of this protein has been linked with a number of disease processes associated with chronic inflammation and fibrosis. Its potential involvement in burns has not yet been investigated. We therefore studied the localization of activin in tissue sections from excised mid- and deep dermal and full thickness cutaneous burn by immunohistochemistry. There was cell-specific temporal expression in tissues with prominent expression from day 4 onwards in lymphocytes and histiocytes and expression from day 8 onwards in reactive fibroblasts and endothelial cells. Immunopositivity over the first 18 days persisted in reactive fibroblasts and lymphocytes although the latter were in most circumstances decreasing in number. These data are consistent with activin A being central to the inflammatory and repair phases occurring in burnt skin and early scar formation. Modulation of activin expression and actions may, therefore, be a target for the management of burns.

    Title Activin-a: a Novel Dendritic Cell-derived Cytokine That Potently Attenuates Cd40 Ligand-specific Cytokine and Chemokine Production.
    Date April 2008
    Journal Blood
    Excerpt

    Activin-A is a transforming growth factor-beta (TGF-beta) superfamily member that plays a pivotal role in many developmental and reproductive processes. It is also involved in neuroprotection, apoptosis of tumor and some immune cells, wound healing, and cancer. Its role as an immune-regulating protein has not previously been described. Here we demonstrate for the first time that activin-A has potent autocrine effects on the capacity of human dendritic cells (DCs) to stimulate immune responses. Human monocyte-derived DCs (MoDCs) and the CD1c(+) and CD123(+) peripheral blood DC populations express both activin-A and the type I and II activin receptors. Furthermore, MoDCs and CD1c(+) myeloid DCs rapidly secrete high levels of activin-A after exposure to bacteria, specific toll-like receptor (TLR) ligands, or CD40 ligand (CD40L). Blocking autocrine activin-A signaling in DCs using its antagonist, follistatin, enhanced DC cytokine (IL-6, IL-10, IL-12p70, and tumor necrosis factor-alpha [TNF-alpha]) and chemokine (IL-8, IP-10, RANTES, and MCP-1) production during CD40L stimulation, but not TLR-4 ligation. Moreover, antagonizing DC-derived activin-A resulted in significantly enhanced expansion of viral antigen-specific effector CD8(+) T cells. These findings establish an immune-regulatory role for activin-A in DCs, highlighting the potential of antagonizing activin-A signaling in vivo to enhance vaccine immunogenicity.

    Title Activin A is a Critical Component of the Inflammatory Response, and Its Binding Protein, Follistatin, Reduces Mortality in Endotoxemia.
    Date November 2007
    Journal Proceedings of the National Academy of Sciences of the United States of America
    Excerpt

    Activin A is a member of the transforming growth factor-beta superfamily, which we have identified as having a role in inflammatory responses. We show that circulating levels of activin increase rapidly after LPS-induced challenge through activation of Toll-like receptor 4 and the key adaptor protein, MyD88. Treatment with the activin-binding protein, follistatin, alters the profiles of TNF, IL-1beta, and IL-6 after LPS stimulation, indicating that activin modulates the release of several key proinflammatory cytokines. Further, mice administered one 10-mug dose of follistatin to block activin effects have increased survival after a lethal dose of LPS, and the circulating levels of activin correlate with survival outcome. These findings demonstrate activin A's crucial role in the inflammatory response and show that blocking its actions by the use of follistatin has significant therapeutic potential to reduce the severity of inflammatory diseases.

    Title Synthesis and Sar of Aminopyrimidines As Novel C-jun N-terminal Kinase (jnk) Inhibitors.
    Date August 2007
    Journal Bioorganic & Medicinal Chemistry Letters
    Excerpt

    The development of a series of novel aminopyrimidines as inhibitors of c-Jun N-terminal kinases is described. The synthesis, in vitro inhibitory values for JNK1, JNK2 and CDK2, and the in vitro inhibitory value for a c-Jun cellular assay are discussed.

    Title The Role of the Activin System in Keloid Pathogenesis.
    Date June 2007
    Journal American Journal of Physiology. Cell Physiology
    Excerpt

    Keloid scars represent a pathological response to cutaneous injury under the regulation of many growth factors. Activin-A, a dimeric protein and a member of the transforming growth factor-beta superfamily, has been shown to regulate various aspects of cell growth and differentiation in the repair of the skin mesenchyme and the epidermis. Thus our aim was to study the role of activin and its antagonist, follistatin, in keloid pathogenesis. Increased mRNA expression for activin was observed in keloid scar tissue by performing RNase protection assay. Immunohistochemistry showed increased localization of both activin-A and follistatin in the basal layer of epidermis of keloid tissue compared with normal tissue. ELISA demonstrated a 29-fold increase in concentration of activin-A and an approximately 5-fold increase in follistatin in conditioned media in keloid fibroblasts compared with normal fibroblasts. Although keloid keratinocytes produced 25% more follistatin than normal keratinocytes, the amounts of activin-A, in contrast, was approximately 77% lower. Proliferation of fibroblasts was stimulated when treated with exogenous activin-A (46% increase in keloids fibroblasts) or following co-culture with hbetaAHaCaT cells (66% increase). Activin-A upregulated key extracellular matrix components, namely collagen, fibronectin, and alpha-smooth muscle actin, in normal and keloid fibroblasts. Co-treatment of follistatin with activin-A blocked the stimulatory effects of activin on extracellular matrix components. These findings emphasize the importance of the activin system in keloid biology and pathogenesis and suggest a possible therapeutic potential of follistatin in the prevention and treatment of keloids.

    Title Interference in Microsphere Flow Cytometric Multiplexed Immunoassays for Human Cytokine Estimation.
    Date April 2007
    Journal Cytokine
    Excerpt

    The present study describes positive and negative interference of human cytokine measurement in multiplexed bead-based immunoassays. Significant differences in measured IL-6 and TNF-alpha values in 30 normal human plasma samples were apparent depending on whether measurements were with a 2-plex assay or embedded in a multiplex of 8-or more cytokine antibody pairs, as well as among the kits of 3-different vendors. Sample diluents containing proprietary blocking ingredients were shown to greatly affect the outcome of measured cytokine values. Additionally, recovery of IL-6 and TNF-alpha from spiked samples suggests significant negative interference from either endogenous antibodies, soluble receptors or anti-cytokine antibodies in 10% and 26% of samples, respectively. While it is evident that multiplexed immunoassays hold great promise for cytokine profiling, there are still important issues needing further study. Especially needed are universally optimized sample diluents, uniformly calibrated standards with mass values, and internal assay controls, which should greatly facilitate intralaboratory accuracy and precision and interlaboratory comparisons of cytokine measurements. Possible causes of interference and remedies are discussed.

    Title Desymmetrization of Diols by a Tandem Oxidation/wittig Olefination Reaction.
    Date March 2007
    Journal Chemical Communications (cambridge, England)
    Excerpt

    Diols are desymmetrized by a tandem oxidation/Wittig olefination to give alpha,beta-unsaturated hydroxy esters without the requirement for protecting group strategies; the alpha,beta-unsaturated hydroxy esters are transformed into dienyl diesters using a second oxidation/Wittig olefination sequence using PCC.

    Title Follistatin is a Candidate Endogenous Negative Regulator of Activin A in Experimental Allergic Asthma.
    Date February 2007
    Journal Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology
    Excerpt

    Activin A is a member of the transforming growth factor-beta superfamily which is directly implicated in airway structural change and inflammation in asthma. In vitro, the biological effects of activin A are neutralized by the soluble binding protein follistatin.

    Title Hypoxia Potentiates Endotoxin-induced Allopregnanolone Concentrations in the Newborn Brain.
    Date January 2007
    Journal Biology of the Neonate
    Excerpt

    Allopregnanolone is a neurosteroid produced in the brain that can alter the excitability of the CNS. Neurosteroids have neuroprotective properties, and their elevation in response to stress may protect the newborn brain following infection or hypoxia. Infection, particularly of the respiratory tract, may lead to episodes of hypoxia. Infection and hypoxia have been identified as factors contributing to neonatal morbidity and mortality.

    Title Activin A Concentrations in Human Cerebrospinal Fluid Are Age-dependent and Elevated in Meningitis.
    Date January 2007
    Journal Journal of the Neurological Sciences
    Excerpt

    Activin A, and its binding protein, follistatin (FS), are expressed in the central nervous system (CNS). We have previously shown elevated concentrations of FS in the cerebrospinal fluid (CSF) of patients with meningitis and increased concentrations of activin A in the CSF of rabbits with bacterial meningitis.

    Title Stimulatory Effects of Lipopolysaccharide on Endothelial Cell Activin and Follistatin.
    Date December 2006
    Journal Molecular and Cellular Endocrinology
    Excerpt

    Activin A and its binding protein, follistatin, are released into the circulation following acute systemic inflammation. In this study, we determined the activin and follistatin response of ovine aortic endothelial cells to lipopolysaccharide (LPS). Exposure to LPS for 1h, mimicking a transient inflammatory event, elicited significant increases in activin betaA subunit mRNA or activin A release, with larger, more prolonged increases evident with continuous exposure. On the other hand, follistatin increases were only evident with prolonged exposure to LPS and following increases in activin A release. While cell-associated activin A increased with LPS exposure, levels were lower than those secreted, whereas the opposite was apparent for follistatin. In summary, our findings suggest that vascular endothelial cells, while capable of releasing activin A and follistatin following inflammatory stimulation, are unlikely to be responsible for the rapid release of activin A in vivo following inflammatory challenge.

    Title Activin a Release into Cerebrospinal Fluid in a Subset of Patients with Severe Traumatic Brain Injury.
    Date November 2006
    Journal Journal of Neurotrauma
    Excerpt

    Activin A is a member of the transforming growth factor-beta superfamily and has been demonstrated to be elevated during inflammation and to have neuroprotective properties following neural insults. In this study, we examined whether traumatic brain injury (TBI) induced a response in activin A or in the concentrations of its binding protein, follistatin. Thirty-nine patients with severe TBI had daily, matched cerebrospinal fluid (CSF) and serum samples collected post-TBI and these were assayed for activin A and follistatin using specific immunoassays. Concentrations of both molecules were assessed relative to a variety of clinical parameters, such as the Glasgow Coma Score, computer tomography classification of TBI, measurement of injury markers, cell metabolism and membrane breakdown products. In about half of the patients, there was a notable increase in CSF activin A concentrations in the first few days post-TBI. There were only minor perturbations in either serum activin or in either CSF or serum follistatin concentrations. The CSF activin A response was not related to any of the common TBI indices, but was strongly correlated with two common markers of brain damage, neuronal specific enolase and S100-beta. Further, activin A levels were also associated with indices of metabolism, such as lactate and pyruvate, excitotoxicity (glutamate) and membrane lipid breakdown products such as glycerol. In one of the two patients who developed a CSF infection, activin A concentrations in CSF became markedly elevated. Thus, some TBI patients have an early release of activin A into the CSF that may result from activation of inflammatory and/or neuroprotective pathways.

    Title Metastatic Potential of Melanomas Defined by Specific Gene Expression Profiles with No Braf Signature.
    Date October 2006
    Journal Pigment Cell Research / Sponsored by the European Society for Pigment Cell Research and the International Pigment Cell Society
    Excerpt

    The molecular biology of metastatic potential in melanoma has been studied many times previously and changes in the expression of many genes have been linked to metastatic behaviour. What is lacking is a systematic characterization of the regulatory relationships between genes whose expression is related to metastatic potential. Such a characterization would produce a molecular taxonomy for melanoma which could feasibly be used to identify epigenetic mechanisms behind changes in metastatic behaviour. To achieve this we carried out three separate DNA microarray analyses on a total of 86 cultures of melanoma. Significantly, multiple testing correction revealed that previous reports describing correlations of gene expression with activating mutations in BRAF or NRAS were incorrect and that no gene expression patterns correlate with the mutation status of these MAPK pathway components. Instead, we identified three different sample cohorts (A, B and C) and found that these cohorts represent melanoma groups of differing metastatic potential. Cohorts A and B were susceptible to transforming growth factor-beta (TGFbeta)-mediated inhibition of proliferation and had low motility. Cohort C was resistant to TGFbeta and demonstrated high motility. Meta-analysis of the data against previous studies linking gene expression and phenotype confirmed that cohorts A and C represent transcription signatures of weakly and strongly metastatic melanomas, respectively. Gene expression co-regulation suggested that signalling via TGFbeta-type and Wnt/beta-catenin pathways underwent considerable change between cohorts. These results suggest a model for the transition from weakly to strongly metastatic melanomas in which TGFbeta-type signalling upregulates genes expressing vasculogenic/extracellular matrix remodelling factors and Wnt signal inhibitors, coinciding with a downregulation of genes downstream of Wnt signalling.

    Title Mycoplasma Pneumoniae Subtype-independent Induction of Proinflammatory Cytokines in Thp-1 Cells.
    Date August 2006
    Journal Microbial Pathogenesis
    Excerpt

    Mycoplasma pneumoniae can be divided into two main subtypes depending on the amino acid sequences of the P1 adhesin and the P65 protein, both located in the attachment organelle. Differences between these subtypes in infectivity, virulence and interaction with host cells have not been extensively studied. Using ELISA to measure released protein and real-time PCR to quantify mRNA, we have demonstrated that both M. pneumoniae subtypes significantly increased tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) and interleukin-8 (IL-8) at comparable levels in THP-1 cells over a 72 h period of time. However, subtype 2 induced a statistically significant increase (P<0.001) in the release of interleukin-1beta at 24 h post-infection compared to subtype 1. These data provide evidence that the induction of proinflammatory cytokine gene and protein expression by M. pneumoniae is not dependent on the infecting subtype.

    Title Dissociation of Angiogenesis and Tumorigenesis in Follistatin- and Activin-expressing Tumors.
    Date July 2006
    Journal Cancer Research
    Excerpt

    The transforming growth factor-beta superfamily member activin and its antagonist, follistatin, act as a pleiotropic growth factor system that controls cell proliferation, differentiation, and apoptosis. Activin inhibits fibroblast growth factor 2-induced sprouting angiogenesis in vitro (spheroidal angiogenesis assay) and in vivo (Matrigel assay). To further study the role of the activin/follistatin system during angiogenesis and tumor progression, activin- and follistatin-expressing R30C mammary carcinoma cells were studied in mouse tumor experiments. Surprisingly, activin-expressing tumors grew much faster than follistatin-expressing tumors although they failed to induce increased angiogenesis (as evidenced by low microvessel density counts). Conversely, follistatin-expressing tumors were much smaller but had a dense network of small-diameter capillaries. Qualitative angioarchitectural analyses (mural cell recruitment, perfusion) revealed no major functional differences of the tumor neovasculature. Analysis of activin- and follistatin-expressing R30C cells identified a cell autonomous role of this system in controlling tumor cell growth. Whereas proliferation of R30C cells was not altered, follistatin-expressing R30C cells had an enhanced susceptibility to undergo apoptosis. These findings in experimental tumors are complemented by an intriguing case report of a human renal cell carcinoma that similarly shows a dissociation of angiogenesis and tumorigenesis during tumor progression. Collectively, the data shed further light into the dichotomous stimulating and inhibiting roles that the activin/follistatin system can exert during angiogenesis and tumor progression. Furthermore, the experiments provide a critical proof-of-principle example for the dissociation of angiogenesis and tumorigenesis, supporting the concept that tumor growth may not be dependent on increased angiogenesis as long as a minimal intratumoral microvessel density is maintained.

    Title Syntheses, Structures, and Photoisomerization of (e)- and (z)-2-tert-butyl-9-(2,2,2-triphenylethylidene)fluorene.
    Date June 2006
    Journal The Journal of Physical Chemistry. A
    Excerpt

    "Sterically geared" 9-(2,2,2-triphenylethylidene)fluorene (1) is of potential interest as a photoactive moiety in molecular devices, and the 2-tert-butyl derivative (6) has been synthesized to investigate photoisomerization. E and Z stereoisomers of 6 were separated and identified by X-ray crystallography. The tert-butyl group does not introduce additional strain, and its close proximity to the trityl group in the Z isomer suggests an attractive van der Waals interaction. The UV spectra of (E)-6 and (Z)-6 are nearly identical, showing absorption bands that are similar to those of fluorene occurring at wavelengths longer than 240 nm. Photoisomerization of 6 was investigated at 266, 280 and 320 nm. Solutions initially containing only (E)-6 or (Z)-6 were irradiated with pulsed laser light, monitoring isomerization by 1H NMR spectroscopy. Negligible photodecomposition was observed when the solutions were agitated by N2 ebullition. Experimental data were fitted to theoretical curves to obtain photoisomerization quantum yields (phi(ZE) and phi(EZ)) ranging from 0.04 to 0.09. This first photoisomerization study of a dibenzofulvene reveals significant quantum yields, despite theoretical prediction of inefficient or negligible isomerization of the parent hydrocarbon, fulvene. Thermal isomerization of 6 at 270 degrees C (t(1/2) = 120 min) was also followed by 1H NMR spectroscopy, resulting in an estimated activation energy (deltaG(double dagger)) of 43 kcal/mol.

    Title Follistatin Attenuates Early Liver Fibrosis: Effects on Hepatic Stellate Cell Activation and Hepatocyte Apoptosis.
    Date January 2006
    Journal American Journal of Physiology. Gastrointestinal and Liver Physiology
    Excerpt

    Activin A, a member of the transforming growth factor-beta superfamily, is constitutively expressed in hepatocytes and regulates liver mass through tonic inhibition of hepatocyte DNA synthesis. Follistatin is the main biological inhibitor of activin bioactivity. These molecules may be involved in hepatic fibrogenesis, although defined roles remain unclear. We studied activin and follistatin gene and protein expression in cultured rat hepatic stellate cells (HSCs) and in rats given CCl4 for 8 wk and examined the effect of follistatin administration on the development of hepatic fibrosis. In activated HSCs, activin mRNA was upregulated with high expression levels, whereas follistatin mRNA expression was unchanged from baseline. Activin A expression in normal lobular hepatocytes redistributed to periseptal hepatocytes and smooth muscle actin-positive HSCs in the fibrotic liver. A 32% reduction in fibrosis, maximal at week 4, occurred in CCl4-exposed rats treated with follistatin. Hepatocyte apoptosis decreased by 87% and was maximal at week 4 during follistatin treatment. In conclusion, activin is produced by activated HSCs in vitro and in vivo. Absence of simultaneous upregulation of follistatin gene expression in HSCs suggests that HSC-derived activin is biologically active and unopposed by follistatin. Our in vivo and in vitro results demonstrate that activin-mediated events contribute to hepatic fibrogenesis and that follistatin attenuates early events in fibrogenesis by constraining HSC proliferation and inhibiting hepatocyte apoptosis.

    Title Activin A: from Sometime Reproductive Factor to Genuine Cytokine.
    Date December 2005
    Journal Veterinary Immunology and Immunopathology
    Excerpt

    The growth factor, activin A, was initially characterized as a putative reproductive hormone but is now known to have many other divergent roles. One of these is during inflammation. Following intravenous injection of bacterial lipopolysaccharide (LPS) into sheep, activin A is released extremely rapidly into the circulation. The release of activin A appears to be independent of fever, prostaglandins or other key proinflammatory cytokines such as TNF-alpha or IL-1beta. While the precise roles and function of this factor in inflammation are yet to be elucidated, the activin response occurs in other mammalian species besides the sheep and elevated activin has been documented for a number of clinical inflammatory conditions. Activin A therefore seems to be part of the regulatory component of the innate immune response.

    Title Furanyl-1,3-thiazol-2-yl and Benzoxazol-5-yl Acetic Acid Derivatives: Novel Classes of Heparanase Inhibitor.
    Date September 2005
    Journal Bioorganic & Medicinal Chemistry Letters
    Excerpt

    Using a furanylthiazole acetic acid as a starting point, a novel series of benzoxazol-5-yl acetic acid derivatives have been identified as heparanase inhibitors. Several compounds possess an IC50 of approximately 200 nM against heparanase, for example, trans 2-[4-[3-(3,4-dichlorophenylamino)-3-oxo-1-propenyl]-2-fluorophenyl]benzoxazol-5-yl acetic acid (16e). Several of the compounds show anti-angiogenic properties. Improvement to the DMPK profile of compounds has provided compounds of potential use in in vivo models.

    Title Cytokine Profiles in the Testes of Rats Treated with Lipopolysaccharide Reveal Localized Suppression of Inflammatory Responses.
    Date June 2005
    Journal American Journal of Physiology. Regulatory, Integrative and Comparative Physiology
    Excerpt

    Evidence indicates that the testis possesses a reduced capacity to mount inflammatory and rejection responses, which undoubtedly contributes to the ongoing survival of the highly immunogenic germ cells. The contribution of local cytokine expression to this condition was investigated in adult male rats treated with lipopolysaccharide to induce inflammation. Cytokine mRNA and protein expression were determined in tissue extracts and fluids by Northern blot analysis, quantitative PCR, or RNAse protection assay and specific ELISAs. Testicular expression of the proinflammatory cytokines, interleukin (IL)-1beta and tumor necrosis factor-alpha was considerably attenuated compared with the liver (control tissue); in contrast, the testicular IL-6 response was enhanced. Expression of IL-10, a type 2 immunoregulatory cytokine, was similar in both testis and liver, whereas the immunoregulatory/anti-inflammatory cytokines transforming growth factor-beta(1) and activin A were constitutively elevated in both normal and inflamed testes. The IL-1beta and transforming growth factor-beta(1) proteins were present principally in their latent (inactive) forms, indicating that enzymic processing is an important control mechanism for these two cytokines within the testis. These data indicate that inflammatory and regulatory cytokine activity is regulated at both transcriptional and posttranslational levels in a testis-specific manner. It is concluded that a novel pattern of suppression of proinflammatory cytokine responses and normal or elevated expression of immunoregulatory cytokines may be responsible for reduced inflammatory responses and enhanced graft survival in the testis. These data have important implications for the understanding and treatment of male autoimmune infertility, testicular inflammation. and carcinogenesis.

    Title Gnrh Agonist Stimulation of the Pituitary-gonadal Axis in Children: Age and Sex Differences in Circulating Inhibin-b and Activin-a.
    Date May 2005
    Journal Human Reproduction (oxford, England)
    Excerpt

    BACKGROUND: Inhibin-B decreases and activin increases FSH secretion in adults. We investigated whether an FSH-inhibin/activin feedback loop exists before or during puberty. METHODS: FSH secretion was stimulated with 10 microg/kg leuprolide acetate (GnRH agonist) in 18 girls, ages 1.0-13.2 years, and 11 boys, ages 8.9-15.2 years, with variations in pubertal development, and in five normal 9- to 10-year-old girls. Blood, obtained at 0, 0.5, 1, 2, 4, 8, 12, 16, 20 and 24 h after GnRH agonist, was analysed for LH, FSH, activin-A, inhibin-A, inhibin-B, follistatin 288 and estradiol/testosterone. RESULTS: FSH increased within 30 min of GnRH agonist administration with a peak greater in girls than boys (P=0.0006). Baseline inhibin-B was greater in boys than girls (P=0.01), while baseline activin-A concentrations were greater in girls. GnRH agonist-stimulated FSH increased inhibin-B in girls by 8 h and in boys by 20 h (P<0.05), but did not affect activin-A. Inhibin-B increases were seen only in girls older than 5 years. CONCLUSIONS: An inhibin-B-FSH feedback loop exists prior to the onset of puberty in girls older than 5 years. Sex differences in activin-A and inhibin-B concentrations may be responsible for sex differences in serum FSH concentrations.

    Title Reciprocal Regulation of Activin A and Inhibin B by Interleukin-1 (il-1) and Follicle-stimulating Hormone (fsh) in Rat Sertoli Cells in Vitro.
    Date May 2005
    Journal The Journal of Endocrinology
    Excerpt

    In several biological systems, the inhibin beta(A) homodimer activin A is stimulated by, and in turn, inhibits the action of interleukin (IL)-1 (both IL-1alpha and IL-1beta) and IL-6. The possibility that a similar regulatory relationship operates within the testis was investigated. Sertoli cells from immature (20-day-old) rats were cultured with human IL-1alpha or IL-1beta, human IL-6 and/or ovine FSH or dibutyryl cAMP. Activin A and the inhibin dimers, inhibin A and inhibin B, were measured by specific ELISA. Immunoreactive inhibin (ir-inhibin) was measured by RIA. Activin/inhibin subunit mRNA expression was measured by quantitative real-time PCR. Both IL-1 isoforms, but not IL-6, stimulated activin A secretion through increased synthesis of beta(A)-subunit mRNA. IL-1 also stimulated activin A secretion by testicular peritubular cells. In contrast to the effect on activin A, IL-1 suppressed inhibin beta(B)-subunit and, to a lesser extent, alpha-subunit mRNA expression, thereby reducing basal and FSH-stimulated inhibin B secretion by the Sertoli cells. Conversely, FSH inhibited basal activin A secretion and antagonised the stimulatory effects of IL-1. Dibutyryl cAMP partially inhibited the action of IL-1 on activin A secretion, but had no significant effect on basal activin A secretion. Secretion of inhibin A was low in all treatment groups. These data demonstrate that IL-1 and FSH/cAMP exert a reciprocal regulation of activin A and inhibin B synthesis and release by the Sertoli cell, and suggest a role for activin A as a potential feedback regulator of IL-1 and IL-6 activity in the testis during normal spermatogenesis and in inflammation.

    Title Effects of Age and Pregnancy on the Circulatory Activin Response of Sheep to Acute Inflammatory Challenge by Lipopolysaccharide.
    Date May 2005
    Journal The Journal of Endocrinology
    Excerpt

    The release of activin A in response to intravenous injection of the bacterial cell-wall component lipopolysaccharide (LPS) was investigated in an ovine model of acute inflammatory challenge in newborn and adult sheep, and in non-pregnant and pregnant ewes. Neonatal lambs (<20 days of age) showed a quantitatively similar response in terms of circulating concentrations of activin A, its binding protein follistatin and the cytokine interleukin-6 compared with adult ewes challenged with an equivalent dose (300 ng/kg bodyweight) of LPS. The fever response and plasma tumour necrosis factor-alpha release in response to LPS, however, were significantly (P < 0.01) less in lambs than in the adult group. Pregnant ewes in the last trimester of gestation had similar responses to LPS, in all aspects measured, compared with their non-pregnant counterparts, apart from an ablated fever response. Although the adult and neonatal sheep responded to LPS, a similar response was not apparent in the fetal circulation, possibly due to a protective effect of the placenta. A 10-fold increase in the dose of LPS (from 300 ng to 3 microg/kg bodyweight) given to neonatal lambs elicited an increase in several cytokine responses measured, with a significant (P< 0.05) increase in follistatin release. In contrast, the amount of activin released by the increased dose of LPS was similar to that invoked by the lower dose. The effect of tolerance to LPS was investigated by giving a second challenge of LPS 5 days after the initial injection. In all animals studied, there was an ablated (P < 0.05) response to the subsequent LPS injection, apart from a similar temperature-response profile. These data provide further evidence that activin A concentrations in the bloodstream are acutely responsive to inflammatory challenge in post-natal life and suggest that the response forms a significant component of the innate immune system.

    Title Activin A and Follistatin in Systemic Inflammation.
    Date May 2005
    Journal Molecular and Cellular Endocrinology
    Excerpt

    Inflammation is a complex process regulated by a cascade of cytokines and growth factors. This review summarizes the emerging evidence implicating activin A and follistatin in the inflammatory process. Our recent studies have highlighted that activin A is released early in the process as part of the circulatory cytokine cascade during acute systemic inflammation. This release occurs concurrently with tumor necrosis factor (TNF)-alpha and prior to that of interleukin (IL)-6 and follistatin. Although, the cellular source(s) of activin A are yet to be established, circulating blood cells and the vascular endothelium are candidates for this rapid release of activin A into the circulation. The release of activin A and follistatin is also observed in the clinical setting, in particular in sepsis. Furthermore activin A is released into cerebrospinal fluid in a model of meningitis in rabbits. The role of activin A in the inflammatory response is poorly understood, however, in vitro data has highlighted that activin A can have both pro- and anti-inflammatory actions on key mediators of the inflammatory response such as TNF-alpha, IL-1beta and IL-6. Furthermore, emerging data would suggest that activin A induction is restricted to certain types of inflammation and its release is dependant upon the inflammatory setting.

    Title Activins, Inhibins and Follistatins in the Large Domestic Species.
    Date February 2005
    Journal Domestic Animal Endocrinology
    Excerpt

    The activins and inhibins are members of the transforming growth factor-beta (TGF-beta) superfamily and, along with follistatin, a high affinity binding protein of activin, form a group of interrelated factors originally isolated for their role in regulating the release of follicle-stimulating hormone (FSH). Knowledge of their function, particularly that of activin, has expanded since being originally isolated, such that they are now regarded as important paracrine regulators in many cellular systems. This review summarizes the biology of these proteins as has been established in the large domestic animals. While the majority of data relate to the pituitary, ovary, uterus/placenta and testis, consideration is also given to emerging roles in inflammatory processes and in non-reproductive tissues or systems.

    Title Sex Differences in Fsh-regulatory Peptides in Pubertal Age Boys and Girls and Effects of Sex Steroid Treatment.
    Date January 2005
    Journal Human Reproduction (oxford, England)
    Excerpt

    BACKGROUND: FSH concentrations are higher in girls than in boys before puberty. We hypothesized that steroid-mediated changes in FSH-regulatory proteins underlie the sex differences in FSH secretion and pubertal timing. METHODS: FSH-regulatory proteins, LH, FSH and sex steroids were measured in five boys, 10 girls, and five girls with Turner syndrome before and during sex steroid treatment (girls, 0.05 mg/day estradiol; boys, 5 mg/day testosterone) for up to 4 weeks. Blood was obtained every 15 min from 20.00 to 08.00 h before and during sex steroid treatment. RESULTS: The mean FSH concentration was higher in girls than in boys (P = 0.0044). Activin-A concentrations were greater (P < 0.0001) and inhibin-B concentrations lower (P < 0.0001) in girls compared with boys. Steroid treatment (i) suppressed LH/FSH concentrations in all subjects; (ii) increased the mean activin-A concentration in all but the Turner girls (P = 0.001); and (iii) decreased inhibin-B concentrations in boys (P = 0.005) but not in girls. Total follistatin and follistatin 288 concentrations did not differ by sex. CONCLUSIONS: Sex steroids regulate circulating activin-A and inhibin-B concentrations in children. The lower inhibin-B and higher activin-A concentrations may explain the higher FSH and earlier onset of puberty in girls.

    Title Persistency of Highly Toxic Coplanar Pcbs in Aquatic Ecosystems: Uptake and Release Kinetics of Coplanar Pcbs in Green-lipped Mussels (perna Viridis Linnaeus).
    Date January 2005
    Journal Environmental Pollution (barking, Essex : 1987)
    Excerpt

    The bioaccumulation potential of three highly toxic coplanar PCB isomers [3,3',4,4'-tetrachlorobiphenyl (T(4)CB); 3,3',4,4',5-pentachlorobiphenyl (P(5)CB); and 3,3',4,4',5,5'-hexachlorobiphenyl (H(6)CB)] was investigated using green-lipped mussels (Perna viridis Linnaeus) as a bioindicator, through a transplantation experiment at two locations in Hong Kong waters. By contrast to the relatively rapid uptake and release of many other PCB isomers, the non-ortho chlorine substituted coplanar PCB congeners exhibited slow uptake and clearance. The kinetic parameters of coplanar PCBs based on lipid weight-related data, and the degree of bioaccumulation based on the proportion of coplanar PCBs in total PCBs in mussels, clearly indicate that coplanar PCBs are highly bioaccumulative in lower organisms. On the assumption that mussels are unlikely to be particularly unusual with respect to their bioaccumulation of coplanar PCBs, it appears most likely that these highly toxic and persistent PCB congeners are concentrated by all aquatic organisms, and may reach higher consumers (including humans) in quantities of toxicological concern.

    Title Cytokines in Mycoplasma Pneumoniae Infections.
    Date December 2004
    Journal Cytokine & Growth Factor Reviews
    Excerpt

    Mycoplasma pneumoniae (M. pneumoniae) is one of the smallest free-living bacteria known. Along with other unique characteristics of this genus, it lacks the typical peptidoglycan cell wall of most eubacteria. Best known for causing tracheobronchitis and atypical pneumonia in humans, this pathogen also causes a number of extrapulmonary syndromes such as meningitis/encephalitis and arthritis. Recent studies also suggest that infection may be associated with chronic conditions such as asthma. Although the mechanisms of M. pneumoniae pathogenesis remain to be elucidated, one important component of M. pneumoniae infections is the induction of proinflammatory and other cytokines in both acute and chronic conditions. In this review, we survey the induction of cytokines by M. pneumoniae in different model systems, and we discuss the possible role of induced cytokines in M. pneumoniae pathogenesis.

    Title Inhibin: Actions and Signalling.
    Date November 2004
    Journal Growth Factors (chur, Switzerland)
    Excerpt

    Inhibin is best known as a reproductive hormone that inhibits release of follicle-stimulating hormone (FSH) from the pituitary gland. Over the last decade or so a number of other biological roles have emerged, putting inhibin more in the class of a growth factor. Despite this, little is known of the signalling pathways for this protein. This minireview summarises the pertinent aspects of inhibin biology and focuses on four potential signalling mechanisms through which inhibin might influence cellular function, namely subunit availability, receptor assembly, co-receptors and signalling through inhibin-specific signalling pathways.

    Title Polychlorinated Biphenyls (pcbs) in Sediments in Hong Kong: a Congener-specific Approach to the Study of Coplanar Pcbs in Aquatic Ecosystems.
    Date October 2004
    Journal Environmental Pollution (barking, Essex : 1987)
    Excerpt

    Patterns of contamination by polychlorinated biphenyls (PCBs) were investigated in fourteen samples of coastal sediments from Hong Kong. Congener-specific analyses revealed nine sediment samples from Junk Bay to contain PCBs at concentrations ranging from 31 to 2200 ng g(-1) dry weight, concentrations generally increasing with distance north in the Bay. By contrast, five sediments from the Tolo area to the north-east of Hong Kong exhibited total PCB levels of only 6.6 to 45 ng g(-) dry weight. The patterns of relative abundance of PCB congeners in the northern Junk Bay sediments suggested the existence of ongoing source(s) of PCBs in this area; biphenyls of lower chlorination were present at high concentration in these samples. Three coplanar PCBs (3', 4, 4'-tetrachlorobiphenyl; 3,3',4,4',5-pentachlorobiphenyl; and 3,3',4,4',5,5'-hexachlorobiphenyl) were found to be present in Junk Bay sediments; these are highly toxic and are cause for concern in aquatic environments. The abundance of the three coplanar PCBs in the sediments studied was similar to that in commercial PCB mixtures, suggesting that these congeners are not enriched over other PCBs by the sediments of coastal ecosystems. It is concluded that the preferential enrichment of coplanar PCBs occurs in the biosphere, rather than in sediments.

    Title Characterisation of the Rapid Release of Activin A Following Acute Lipopolysaccharide Challenge in the Ewe.
    Date September 2004
    Journal The Journal of Endocrinology
    Excerpt

    A series of experiments were conducted in adult ewes to delineate the release profile of activin A and its relationship to other cytokines following an i.v. injection of the bacterial cell wall component, lipopolysaccharide (LPS). Following this challenge, plasma activin A increased rapidly and appeared to be released in a biphasic manner, slightly preceding the release of tumour necrosis factor-alpha (TNFalpha) and before elevation of interleukin (IL)-6 and follistatin levels. The concentration of activin A was correlated with body temperature during the response to LPS. A second experiment compared cytokine concentrations in matched blood and cerebrospinal fluid (CSF) samples. This revealed that activin A was not released centrally in the CSF following a peripheral LPS injection, nor was TNFalpha or the activin binding protein, follistatin, but IL-6 showed a robust elevation. In a third experiment, the stimulus for activin A release was examined by blocking prostaglandin synthesis. Flurbiprofen, a prostaglandin synthesis inhibitor, effectively attenuated the fever response to LPS and partly inhibited cortisol release, but the cytokine profiles were unaffected. Finally, the bioactivity of TNFalpha and/or IL-1 was blocked using soluble receptor antagonists. These treatments did not affect the initial release of activin A, but blockade of TNFalpha depressed the second activin peak. These studies define more rigorously the release of activin A into the circulation following acute inflammatory challenge. The response is rapid and probably biphasic, independent of prostaglandin- mediated pathways and does not depend upon stimulation by TNFalpha or IL-1. The data suggest that activin A release is an early event in the inflammatory cascade following the interaction of LPS with its cellular receptor.

    Title Increased Mrna Expression of Kynurenine Pathway Enzymes in Human Placentae Exposed to Bacterial Endotoxin.
    Date September 2004
    Journal Advances in Experimental Medicine and Biology
    Excerpt

    Intra-amniotic bacterial infection is a major risk factor for cerebral impairment in infants that are born pre-term however, the causal pathways are largely unknown. Whether placental derived, neuroactive kynurenine metabolites play any role in fetal cerebral damage during episodes of intra-amniotic infection is presently unknown. In this preliminary study, we explored if kynurenine metabolites may be involved, examining if mRNAs of enzymes involved in tryptophan catabolism through the kynurenine pathway (KP) were expressed in the placenta and if their expression was co-ordinately altered with exposure to bacterial infection. We found that placentae from healthy women at term and those with clinical signs of amniotic fluid bacterial infection pre-term expressed mRNAs of the KP enzymes, with higher expression overall in the infected group. Significant increases in indoleamine 2,3-dioxygenase (IDO), tryptophan dioxygenase (TDO) and kynureninase (KYNase) expression were detected in association with infection. These findings suggest that tryptophan may be constitutively degraded through the KP in the human placenta. Whether higher concentrations of placental derived kynurenine metabolites enter the fetus during episodes of infection and their physiological roles if any remains to be elucidated.

    Title Effect of Heparin Administration to Sheep on the Release Profiles of Circulating Activin A and Follistatin.
    Date July 2004
    Journal The Journal of Endocrinology
    Excerpt

    Activin A and follistatin are normally present in relatively low amounts in the circulation. Heparin administration elicits a rapid and robust release of these proteins, although this phenomenon is poorly defined. In the present studies, the response to heparin administration was evaluated in the plasma of adult ewes in terms of whether it was dose-dependent, could be neutralized, was responsive to multiple stimulation, and the nature of the activin A and follistatin released. Activin A and follistatin were rapidly released by heparin in a dose-dependent manner (25, 100 or 250 IU/kg), with differences in the response as adjudged by peak concentration, timing of the peak and area under the curve. The heparin response could be blocked by pretreatment with protamine; conversely protamine injection alone (2 mg/kg) elicited release of follistatin but not activin A. Repeat administration of heparin at three-hourly intervals resulted in activin and follistatin responses to each injection, but each subsequent stimulation increased and extended the responses, consistent with saturation of the heparin clearance mechanism. Size exclusion chromatography of plasma samples confirmed that the majority of activin and follistatin released by heparin was a complex, whereas follistatin released by protamine was unbound. These data are consistent with a large pool of activin A and follistatin resident on extracellular matrices, with the rapid response implicating the vascular endothelium as the prime site of release following administration of these commonly used anticoagulant therapies.

    Title Mussels As Bioindicators of Pcb Pollution: a Case Study on Uptake and Release of Pcb Isomers and Congeners in Green-lipped Mussels (perna Viridis) in Hong Kong Waters.
    Date June 2004
    Journal Environmental Pollution (barking, Essex : 1987)
    Excerpt

    The uptake and release of PCB isomers and congeners were examined in green-lipped mussels (Perna viridis) through a transplantation experiment in two locations in Hong Kong waters. Rapid rates of uptake and release of relatively less lipophilic lower-chlorinated PCBs were observed in the mussels, indicating that the primary mechanism of bioaccumulation of lipophilic pollutants in P. viridis complies with the concept of equilibrium partitioning. Thus, data for contaminant concentrations are most appropriately based upon lipid weights of samples when using mussels as bioindicators of aquatic PCB pollution. Considering the kinetic parameters of PCBs based on lipid weight-related data, it is concluded that P. viridis has the ability to respond rapidly to changes in ambient levels of PCBs. This is significant in determining the usefulness and limitations of mussels as bioindicators for monitoring programmes investigating aquatic pollution by PCBs.

    Title Role of Mass Spectrometry in the Purification of Peptides and Proteins.
    Date February 2004
    Journal Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
    Excerpt

    Experiments were carried out to evaluate the fractionation of proteins and peptides according to mass. Model mixtures were separated by either reversed-phase or ion-exchange chromatography with mass spectrometry-compatible mobile phase additives. Fraction collection was triggered by the mass/charge ratio of each one of the components of the mixture. Chromatography was additionally monitored with a UV-Vis detector in order to compare the new technique with generally accepted in separations. The results indicated that adequate purification is achieved by this new technique. Fraction collection triggered by changes in the mass/charge ratio reduces sample handling and analysis time. This study demonstrates the utility of mass-directed fractionation of peptides and proteins when mass spectrometry-compatible mobile phase additives are used.

    Title Fetal Responses to Maternal and Intra-amniotic Lipopolysaccharide Administration in Sheep.
    Date December 2003
    Journal Biology of Reproduction
    Excerpt

    A link between intrauterine infection and premature labor is widely accepted, yet the fetal inflammatory responses to such infections are not well understood. Our aim was to use a sheep model in which an inflammatory state was induced by lipopolysaccharide (LPS) administration during pregnancy to the maternal systemic, intra-amniotic or extra-amniotic compartments. Fetal and maternal blood gases and uterine electromyographic activity along with fetal and maternal circulating concentrations of prostaglandins PGE2 and PGFM, cortisol, and interleukin-6 were determined. Maternal systemic LPS treatment resulted in mild maternal hypoxemia, a rise in temperature, greater fetal hypoxemia, and a marked rise in fetal cortisol and PGE2 concentrations that persisted for 48 h. Intra-amniotic administration of LPS at doses higher than those used systemically caused an increase in fetal cortisol and PGE2 concentrations as well as a rise in uterine activity, but these were lesser in magnitude. Extra-amniotic LPS administration caused no overt fetal or maternal inflammatory responses. We conclude that maternal LPS treatment markedly elevated fetal cortisol and PGE2 concentrations. This may be a potential protective mechanism that aids the fetus in the event of premature delivery. The attenuated fetal response to intra-amniotic LPS treatment, despite the much higher dose used, may support a role for the amniotic fluid in protecting the fetus from endotoxin exposure during pregnancy.

    Title Regulation of Ovarian Function by the Tgf-beta Superfamily and Follistatin.
    Date November 2003
    Journal Reproduction (cambridge, England)
    Excerpt

    The role of follistatin as an activin-binding protein has dominated the study of this molecule for the last 10 years. However, there is emerging evidence that follistatin has a role in modulating the biology of other members of the transforming growth factor beta (TGF-beta) superfamily. This review summarizes the current concepts encompassing follistatin biochemistry as well as molecules with which it is functionally associated. Moreover, the importance of the two follistatin isoforms (follistatin-288 and follistatin-315) is discussed with particular emphasis on the regulation of the ovary. In addition to activin, this review discusses the functions of other members of the TGF-beta superfamily, for example growth differentiation factor 9 (GDF-9), bone morphogenetic protein 15 (BMP-15), BMP-6, BMP-4 and BMP-7, in the ovary, and the potential interactions between follistatin and these growth factors. The complex network of TGF-beta superfamily growth factor members involved in the modulation of ovarian function and the interactions of follistatin with these proteins is highlighted.

    Title Activin Betac-subunit Heterodimers Provide a New Mechanism of Regulating Activin Levels in the Prostate.
    Date October 2003
    Journal Endocrinology
    Excerpt

    Activins are formed by dimerization of beta-subunits and, as members of the TGF-beta superfamily, have diverse roles as potent growth and differentiation factors. As the biological function of the activin C homodimer (betaC-betaC) is unknown, we sought to compare activin A (betaA-betaA), B (betaB-betaB), and C homodimer bioactivities and to investigate the consequences of activin betaC-subunit overexpression in prostate tumor cells. Exogenous activin A and B homodimers inhibited cell growth and activated activin-responsive promoters. In contrast, the activin C homodimer was unable to elicit these responses. We previously showed that the activin betaC-subunit heterodimerized with activin betaA in vitro to form activin AC. Therefore, we hypothesize that the activin betaC-subunit regulates the levels of bioactive activin A by the formation of activin AC heterodimers. To test this hypothesis, we measured activin AC heterodimer production using a novel specific two-site ELISA that we developed for this purpose. In the PC3 human prostate tumor cell line, activin betaC-subunit overexpression increased activin AC heterodimer levels, concomitantly reduced activin A levels, and decreased activin signaling. Overall, these data are consistent with a role for the activin betaC-subunit as a regulatory mechanism to reduce activin A secretion via intracellular heterodimerization.

    Title Differential Responses of Human Umbilical and Coronary Artery Endothelial Cells to Apoptosis.
    Date October 2003
    Journal Endothelium : Journal of Endothelial Cell Research
    Excerpt

    Apoptosis, or programmed cell death, plays an important role not only in normal homeostasis but is increasingly implicated in a number of pathological processes. The contribution of apoptosis in the pathophysiology of coronary artery disease has been suggested. In this study, the authors compared the effects of inductive and suppressive signals as well as the two combined apoptotic signals during the early stages of apoptosis using two types of human endothelial cells isolated from the umbilical vein (HUVECs) and from the coronary artery (HCAECs) vascular beds as study targets. The authors demonstrated that HUVECs were more susceptible than HCAECs to apoptosis, as measured by a 2.5-fold increase in caspase-3 activity, which in turn may suggest a different pattern in association to the apoptotic mechanism within each cell type. Under inducing conditions of apoptosis, a significant increase in tissue factor (TF) expression at both the mRNA and protein level in HUVECs compared to the level of TF expression in HCAECs was observed. These different responses of endothelial cell types potentially indicate unequal susceptibility to apoptosis, depending on their vascular bed origins, and emphasize the importance of cell-type and -origin considerations when selecting a study model of apoptosis and pathophysiology.

    Title Increased Activin Levels in Cerebrospinal Fluid of Rabbits with Bacterial Meningitis Are Associated with Activation of Microglia.
    Date August 2003
    Journal Journal of Neurochemistry
    Excerpt

    Activin, a member of the transforming growth factor superfamily, is upregulated in a number of inflammatory episodes such as septicemia and rheumatoid arthritis. In the CNS, activin has been predominantly assessed in terms of a neuroprotective role. In this report we characterized the activin response in the CNS in a rabbit model of meningitis. In normal animals, cerebrospinal fluid (CSF) activin levels were higher than those in serum, indicating an intracranial secretion of this cytokine. Following intracisternal inoculation with Streptococcus pneumoniae, activin in CSF was unchanged for the first 12 h and then rose progressively; levels were increased approximately 15-fold within 24 h. Activin levels were correlated positively with CSF protein content and with the number of apoptotic neurons in the dentate gyrus. No apparent correlation was observed between CSF activin concentrations and bacterial titer, lactate concentrations or leukocyte density. Using immunohistochemistry, activin staining was localized to epithelial cells of the choroid plexus, cortical neurons and the CA3 region of the hippocampus, with similar staining intensities in both normal and meningitic brains. However, in meningitic brains there was also strong staining in activated microglia and infiltrating macrophages. Taken together, these results demonstrate that activin forms part of the CNS response to immune challenge and may be an important mediator to modulate inflammatory processes in the brain.

    Title Bond Angle Versus Torsional Deformation in an Overcrowded Alkene: 9-(2,2,2-triphenylethylidene)fluorene.
    Date July 2003
    Journal Chemistry (weinheim an Der Bergstrasse, Germany)
    Excerpt

    Competition between bond angle and torsional strain in sterically crowded alkenes generally causes twisting in tetrasubstituted alkenes, while most structurally characterized trisubstituted alkenes are planar. To investigate structural effects of steric repulsion between a planar aromatic ring and a vicinal triphenylmethyl (trityl) group, 9-(2,2,2-triphenylethylidene)fluorene (1 a) was synthesized by reaction of 9-bromomethylenefluorene with triphenylmethyllithium. For comparison with a less strained analogue, 9-ethylidenefluorene (1 b) was prepared by reaction of fluorenone with ethylmagnesium bromide. The X-ray crystal structures show that the difference between bond angles at the 9-fluorenyl carbon atom is much larger for 1 a (12.9 degrees) than 1 b (2.6 degrees). Bond angle and torsional deformations were compared theoretically (HF/6-31+G*) with the tert-butyl analogue (1 c), 1,2,2-tri-tert-butylethene (7), and 2,4,4-trimethyl-2-pentene (8) and crystallographically with six known 1,1-diaryl-2-tert-alkylethenes (2). The trisubstituted alkenes formed three groups with 1) large angle distortion with moderate twisting (1 a, 1 b, and 7), 2) moderate bending with a large range of torsional angles (2), and 3) little bending or twisting (1 b and 8). For the entire series, there appears to be a delicate balance between angle and torsional deformation, but twisting appears to produce smaller relief from steric strain than angle bending. In the crystallographically characterized trisubstituted alkenes, the choice between the two is mainly determined by more subtle packing forces.

    Title Interleukin-1beta Responses to Mycoplasma Pneumoniae Infection Are Cell-type Specific.
    Date June 2003
    Journal Microbial Pathogenesis
    Excerpt

    Interleukin-1beta (IL-1beta) is a major proinflammatory cytokine that is involved in many important cellular functions such as proliferation, differentiation, and activation of different cell types. Its mature form is released from the cells in response to various bacterial and viral infections, and it plays a significant role in host defense. Mycoplasma pneumoniae is a small bacterium without a cell wall that causes tracheobronchitis and atypical pneumonia in humans following attachment to respiratory epithelium, as well as extrapulmonary infections. Very little is known about the role of cytokines in pathogenesis or the response of target cells to M.pneumoniae attachment. The purpose of this study was to investigate the ability of M. pneumoniae to induce IL-1beta in human lung epithelial carcinoma A549 and in human monocytic U937 cell lines. Following M. pneumoniae infection, both IL-1beta mRNA and protein were induced in A549 cells vs. no induction in uninfected cells; however, the protein remained inside the A549 cells. Similarly, M. pneumoniae infection strongly increased mRNA and extracellular protein levels in U937 cells, which unlike A549 cells did exhibit baseline constitutive levels. De novo IL-1beta protein expression was verified by cycloheximide studies. M. pneumoniae infection did not affect constitutive caspase-1 mRNA or protein levels in either cell line. Reduced caspase-1 activity in A549 cell lysates suggests the presence of an endogenous caspase-1 inhibitory component in the A549 cells. These collective data confirm previous studies that show that M. pneumoniae is a potent inducer of cytokines following adherence to host target cells, and establish that IL-1beta release in response to M. pneumoniae infection is cell-type specific, thus emphasizing the importance of carefully considering multiple cell types in M. pneumoniae pathogenesis studies involving both immune cells and cytokine release patterns.

    Title A Periconceptional Nutritional Origin for Noninfectious Preterm Birth.
    Date May 2003
    Journal Science (new York, N.y.)
    Title Inhibins, Activins and Follistatin in Reproduction.
    Date May 2003
    Journal Human Reproduction Update
    Excerpt

    The regulation of reproductive processes involves a complex network of communication systems between the brain, endocrine organs, the gonads and other reproductive tissues. Classically, our understanding has focused on the role of endocrine hormones, but more recently interest has also dwelt on the paracrine and autocrine regulation of these cell systems. In this review, the structure and physiology of the inhibins, activins and follistatin are discussed in terms of the evidence supporting their role as endocrine hormones, and how they might function as paracrine factors within the pituitary, gonad and associated tissues. With the advent of more specific techniques and assays for their measurement, the potential of inhibins, activins and follistatin as clinical markers of reproductive function and in the screening of various pathologies is also evaluated.

    Title Induction of Proinflammatory Cytokines in Human Lung Epithelial Cells During Chlamydia Pneumoniae Infection.
    Date February 2003
    Journal Infection and Immunity
    Excerpt

    Chlamydia pneumoniae is an obligate intracellular human pathogen that causes acute respiratory diseases such as pneumonia and bronchitis. Previous studies have established that C. pneumoniae can induce cytokines in mouse and/or human cells, but little information is available on the cytokine response of respiratory epithelial cells, a first line of infection. In this study, heparin treatment of C. pneumoniae significantly reduced its ability to induce interleukin 8 (IL-8) and tumor necrosis factor alpha (TNF-alpha) mRNA in human lung carcinoma cells, indicating that cytadherence is an important early stimulus for induction of proinflammatory mediators. Although the IL-8, gamma interferon, and TNF-alpha message was consistently induced by infection of A549 cells not treated with heparin, only an elevation of IL-8 protein was detected in A549 supernatants. A549 IL-beta and IL-6 mRNA and supernatant protein profiles were not significantly changed by infection. Heat or UV inactivation of C. pneumoniae only partially reduced the cytokine response, and inhibition of C. pneumoniae protein or DNA synthesis did not affect its ability to induce cytokine gene expression. To prevent stress-induced cytokine release by the A549 cells, centrifugation was not utilized for infection experiments. These experiments establish the importance of cytadherence in cytokine release by cells of respiratory epithelial origin and suggest that further work in the area of cytokine mediators is warranted to gain valuable pathogenic and therapeutic insights.

    Title Centrifugation of Human Lung Epithelial Carcinoma A549 Cells Up-regulates Interleukin-1beta Gene Expression.
    Date February 2003
    Journal Clinical and Diagnostic Laboratory Immunology
    Title Neuroendocrine Control of Follicle-stimulating Hormone (fsh) Secretion: Ii. Is Follistatin-induced Suppression of Fsh Secretion Mediated Via Changes in Activin Availability and Does It Involve Changes in Gonadotropin-releasing Hormone Secretion?
    Date September 2002
    Journal Biology of Reproduction
    Excerpt

    The objective of the present study was to determine to what extent activin participates in setting the level of FSH secretion and if this regulation includes mediation via changes in GnRH secretion. We administered follistatin, the high-affinity binding protein for activin, to five ovariectomized sheep; we reasoned that the resultant binding of follistatin to activin should lower activin bioavailability and FSH secretion. Hypophyseal portal and peripheral blood samples were collected simultaneously at 10-min intervals for 18 h to measure GnRH, LH, FSH, and both activin-free and total follistatin. Six hours into collection, each ewe received 150 microg/kg i.v. of recombinant human follistatin-288. A week later, the same ewes were subjected to a second series of blood collections of similar length (time control). The FSH levels in pituitary portal blood were approximately 8-fold higher than those in the peripheral circulation. The FSH secretory patterns changed minimally during the time-control period. In contrast, follistatin had profound suppressive effects on FSH secretion. Maximal FSH suppression after FS-288 administration occurred at 5-6 h in the pituitary portal (65% suppression) and 9-10 h in the peripheral (48% suppression) circulation. Follistatin had no effect on GnRH or LH secretory patterns. Disappearance of total follistatin (i.e., free follistatin plus activin-bound follistatin) from the circulation was slower (P < 0.05) than that of free follistatin alone, suggesting that some of the follistatin was complexed with circulating activin, thus reducing the bioavailability of activin. The slower clearance of total follistatin and the lack of follistatin effects on GnRH secretion suggest that changes in activin bioavailability dictate the level of pituitary FSH secretion and that this is a pituitary-specific effect.

    Title Regulation of Proinflammatory Cytokines in Human Lung Epithelial Cells Infected with Mycoplasma Pneumoniae.
    Date July 2002
    Journal Infection and Immunity
    Excerpt

    Mycoplasma pneumoniae is a small bacterium without a cell wall that causes tracheobronchitis and atypical pneumonia in humans. It has also been associated with chronic conditions, such as arthritis, and extrapulmonary complications, such as encephalitis. Although the interaction of mycoplasmas with respiratory epithelial cells is a critical early phase of pathogenesis, little is known about the cascade of events initiated by infection of respiratory epithelial cells by mycoplasmas. Previous studies have shown that M. pneumoniae can induce proinflammatory cytokines in several different study systems including cultured murine and human monocytes. In this study, we demonstrate that M. pneumoniae infection also induces proinflammatory cytokine expression in A549 human lung carcinoma cells. Infection of A549 cells resulted in increased levels of interleukin-8 (IL-8) and tumor necrosis factor alpha mRNA, and both proteins were secreted into culture medium. IL-1 beta mRNA also increased after infection and IL-1 beta protein was synthesized, but it remained intracellular. In contrast, levels of IL-6 and gamma interferon mRNA and protein remained unchanged or undetectable. Using protease digestion and antibody blocking methods, we found that M. pneumoniae cytoadherence is important for the induction of cytokines. On the other hand, while M. pneumoniae protein synthesis and DNA synthesis do not appear to be prerequisites for the induction of cytokine gene expression, A549 cellular de novo protein synthesis is responsible for the increased cytokine protein levels. These results suggest a novel role for lung epithelial cells in the pathogenesis of M. pneumoniae infection and provide a better understanding of M. pneumoniae pathology at the cellular level.

    Title Activated Protein C Induction of Mcp-1 in Human Endothelial Cells: a Possible Role for Endothelial Cell Nitric Oxide Synthase.
    Date March 2002
    Journal Thrombosis Research
    Excerpt

    Classically, activated protein C (APC) of the protein C/protein S anticoagulant pathway has functioned not only to inactivate the procoagulant factors Va and VIIIa but also to inhibit the activity of plasminogen activator inhibitor-1 (PAI-1). More recent data have suggested that the protein C/protein S pathway may serve as a physiological link between coagulation and inflammation. This APC pathway link was proposed because of observations showing that APC could both modulate the effects of cytokines and block neutrophil activation. As a further extension of the effect(s) of APC on cytokines, we found that APC, at the equivalent physiological protein C concentration of 4 microg/ml, significantly upregulated monocyte chemotactic protein-1 (MCP-1) RNA in human umbilical vein endothelial cells (HUVECs), as indicated by a ribonuclease protection assay (RPA) at 3 and 6 h with a return to near basal levels by 24 h. ELISA determinations demonstrated that 4 microg/ml of APC induced a significant (P=.0001) increase in MCP-1 protein production over basal levels within a 24-h period. At the same concentration, APC downregulated endothelial cell nitric oxide synthase (eNOS) RNA. Downregulation first became apparent at 6 h and continued through 48 h of culture. This downregulation was concentration dependent over a range of 1.3-12 microg/ml, and there was no effect on cell viability within this range. In support of other studies, we also found that exogenously added nitric oxide (NO) inhibited MCP-1 production. These data suggest that APC may induce MCP-1 through the inhibition of eNOS.

    Title Inhibins, Activins and Follistatin: Actions on the Testis.
    Date December 2001
    Journal Molecular and Cellular Endocrinology
    Excerpt

    While the early studies of the inhibins, activins and follistatins concentrated on their role as endocrine regulators of FSH secretion, recent data has emphasized the local actions of the activins and follistatin. Inhibin, through its capacity to suppress FSH secretion can modulate numerous processes within the testis. However, to date, evidence to support a local role for inhibin is limited. In contrast, activin and its binding protein follistatin are produced by a large number of cell-types within the testis raising the possibility of a range of paracrine and autocrine actions. These include the modulation of androgen production, influence on the proliferation of Sertoli cells and germ cells as well as the capacity to influence the structural and functional features of mitochondria within germ cells. Some of these actions are carefully controlled in a temporal relationship during the development of testicular function in the rat in which there is no separation in time between birth and the onset of spermatogenesis. Given the range of actions of activin in different cell-types, recognition of systems that are designed to modulate its actions are crucial in enhancing our understanding of how these many roles can be compartmentalized.

    Title Evidence for Activin A and Follistatin Involvement in the Systemic Inflammatory Response.
    Date December 2001
    Journal Molecular and Cellular Endocrinology
    Excerpt

    The inflammatory cascade is a multifactorial process regulated by interwoven cytokine and growth factor networks. This review summarizes the emerging evidence that implicate activin A and follistatin in inflammatory processes. Our recent studies have determined that activin A is released early in the cascade of circulatory cytokines during systemic inflammatory episodes, roughly coincident with tumour necrosis factor (TNF)-alpha and before interleukin (IL)-6 and follistatin. The source(s) of this activin A are not yet established, but prime candidates are monocytes/macrophages, other immune cell types or vascular endothelial cells. Clinical data are limited, but activin beta(A) subunit mRNA or activin A protein is elevated in inflammatory bowel diseases and inflammatory arthropathies, and circulating concentrations of follistatin are elevated in patients with sepsis. In more mechanistic approaches, in vitro studies show that activin A can have both pro- and anti-inflammatory actions on key inflammatory mediators such as TNFalpha, IL-1beta and IL-6. Furthermore, there is emerging understanding of how the intracellular signaling pathway for activin A, incorporating Smads, may interact with and be modulated by other key regulatory cytokines and growth factors.

    Title Characterization of Serum Activin-a and Follistatin and Their Relation to Virological and Histological Determinants in Chronic Viral Hepatitis.
    Date October 2001
    Journal Journal of Hepatology
    Excerpt

    Hepatocyte proliferation in viral hepatitis is regulated by a number of growth factors. Activin-A inhibits hepatocyte DNA synthesis while follistatin, a potent activin-A antagonist, promotes liver regeneration. We report the first study of activin-A and follistatin in human viral hepatitis. Sera from 15 normal subjects, 22 hepatitis B and 47 hepatitis C patients were analysed for activin-A and follistatin and correlated with serological and histological markers of liver injury and with specific immunohistochemistry.

    Title Expression of Chemokine by Human Coronary-artery and Umbilical-vein Endothelial Cells and Its Regulation by Inflammatory Cytokines.
    Date September 2001
    Journal Coronary Artery Disease
    Excerpt

    BACKGROUND: Activation of the endothelium is a critical event in the process of inflammation and is associated with the production of chemokines. OBJECTIVE: To evaluate the proinflammatory cytokine-induced chemokine repertoire of human coronary-artery endothelial cells (HCAEC) both at the messenger RNA (mRNA) level and at protein level in direct comparison with that of human umbilical-vein endothelial cells (HUVEC). METHODS: Human coronary-artery and human umbilical-vein endothelial cells were obtained commercially and experimental data were derived from cell cultures between passage levels 3 through 6. Supernatant fluids from cytokine [tumor necrosis factor-alpha (TNF-alpha), interleukin-1-alpha, and anti-TNF R55] stimulated endothelial cell cultures were used to study chemokine release. Sandwiched ELISA assays, obtained commercially, were used to estimate cell culture supernatant fluid levels of the selected chemokines: monocytic chemotactic protein-1, regulated upon activated normal T cells expressed and secreted, interleukin-8, transforming growth factor-beta-2 (TGF-beta2), and gamma interferon protein-10. Expression of messenger RNA was determined using selected labeled riboprobes (32P UTP) in a ribonuclease protection assay using total cellular mRNA. RESULTS: Upon in-vitro stimulation with TNF-alpha and interleukin-1-alpha, production of regulated-upon-activated-normal-T-cells expressed and secreted (RANTES) protein by HCAEC was significantly increased relative to that by HUVEC, the greatest effect being found with interleukin-1-alpha. The opposite effect, however, was noted for levels of monocytic-chemotactic-protein-1 protein, which were detected in HUVEC at significantly higher levels than they were in HCAEC challenged by those cytokines. Production of gamma interferon-inducible protein-10 (gammaIP-10) by HUVEC was induced by TNF-alpha and interleukin-1-alpha, whereas only a modest induction by interleukin-1-alpha was seen in HCAEC. TGF-beta-2 protein was constitutively expressed in HCAEC but not in HUVEC. Expression of mRNA was analyzed by the ribonuclease-protectionassay. RANTES mRNA was expressed in HCAEC from 3 h through 48 h after treatment with TNF-alpha, whereas only a modest induction of RANTES was expressed in HUVEC 24 h and 48 h after treatment with TNF-alpha. Monocytic-chemotactic-protein-1 mRNA was constitutively expressed by both types of cell, but the basal levels in HCAEC was significantly higher than in HUVEC. HCAEC constitutively expressed both TGF-beta-1 and TGF-beta-2 mRNA, whereas HUVEC constitutively expressed TGF-beta-1 only. CONCLUSION: Our data indicate that HCAEC and HUVEC express chemokines differently, which could contribute to or influence site-specific recruitment of subsets of leukocytes.

    Title Mixed-mode Solid-phase Extraction and Cleanup Procedures for the Liquid Chromatographic Determination of Thiabendazole and Carbendazim in Fruit Juices.
    Date August 2001
    Journal Journal of Aoac International
    Excerpt

    Solid-phase extraction (SPE) procedures were developed for rapid cleanup and determination of thiabendazole and carbendazim in orange, apple, and grape juices. Samples were prepared by using an SPE cartridge containing a mixed-mode sorbent with both reversed-phase and strong cation-exchange chemistries. Analysis was by liquid chromatography with photodiode-array UV detection. Orange juice was analyzed by mixed-mode cation-exchange extraction with reversed-phase cleanup; the other juices were analyzed by reversed-phase extraction with cation-exchange cleanup. Recoveries >80% for carbendazim and >90% for thiabendazole. Quantitation limits were 20 microg/L for both analytes.

    Title New Developments in the Biology of Inhibins, Activins and Follistatins.
    Date July 2001
    Journal Trends in Endocrinology and Metabolism: Tem
    Title The Sac-4 Gene of Neisseria Gonorrhoeae and Co-existing Chlamydial Infection.
    Date February 2001
    Journal Sexually Transmitted Infections
    Excerpt

    Recently, the sac-4 gene in Neisseria gonorrhoeae was postulated to increase the risk of developing mixed gonococcal and chlamydial infection. The aims of this study were to determine the frequency of the sac-4 gene in a larger sample of isolates of different serovars and to assess the prevalence of sac-4 in gonococcal isolates from patients with and without coexisting chlamydial infection.

    Title Inducible Nitric Oxide Synthase in the Rat Testis: Evidence for Potential Roles in Both Normal Function and Inflammation-mediated Infertility.
    Date December 2000
    Journal Biology of Reproduction
    Excerpt

    In vitro data have indicated that nitric oxide (NO) inhibits Leydig cell testosterone production, suggesting that NO may play a role in the suppression of steroidogenesis and spermatogenic function during inflammation. Consequently, we investigated expression of the inflammation-inducible isoform of NO synthase (iNOS) in the inflamed adult rat testis and the ability of a broad-spectrum inhibitor of NO production, L-nitro-L-arginine methyl ester, to prevent Leydig cell dysfunction during inflammation. Unexpectedly, immunohistochemical and mRNA data established that iNOS is expressed constitutively in Leydig cells and in a stage-specific manner in Sertoli, peritubular, and spermatogenic cells in the normal testis. Expression was increased in a dose-dependent manner in all these cell types during lipopolysaccharide (LPS)-induced inflammation. In noninflamed testes, treatment with the NO synthase inhibitor reduced testicular interstitial fluid formation and testosterone production without any effect on serum LH levels. Administration of the inhibitor did not prevent the suppression of testicular interstitial fluid and testosterone production that occurs within 6 h after LPS treatment. Collectively, these data indicate a novel role for iNOS in autocrine or paracrine regulation of the testicular vasculature, Leydig cell steroidogenesis, and spermatogenesis in the normal testis. The data suggest that increased NO is not the major cause of acute Leydig cell dysfunction in the LPS-treated inflammation model, although a role for NO in this process cannot be excluded, particularly at other time points. Moreover, up-regulation of iNOS may contribute to the seminiferous epithelium damage caused by LPS-induced inflammation.

    Title The Role of the T-pa I/d and Pai-1 4g/5g Polymorphisms in African-american Adults with a Diagnosis of Myocardial Infarction or Venous Thromboembolism.
    Date November 2000
    Journal Thrombosis Research
    Excerpt

    To determine whether or not the PAI-1 4G/5G and t-PA I/D polymorphisms in African-Americans were linked to cardiovascular disease, the association of these polymorphisms to disease expression was analyzed in a recently completed case-control study of myocardial infarction or venous thromboembolism among African-Americans. All African-Americans patients with a history of venous thromboembolism attending an anticoagulant clinic, and patients with a history of a MI attending a cardiology clinic at a large local urban public hospital were eligible for inclusion as cases in the study. In this study it was observed that there was a statistically significant association between the D allele of the t-PA I/D polymorphism and venous thromboembolism and a nonsignificant association between the D allele and myocardial infarction among African-Americans. t-PA antigen levels were statistically significantly higher among both myocardial infarction and venous thromboembolism cases compared with control subjects. The genotypes were unrelated to t-PA plasma levels. There was no association between either myocardial infarction or venous thromboembolism and the 4G/5G PAI-1 genotype. It was also found that genotype frequencies for both PAI-1 4G/5G and t-PA I/D polymorphisms in African-American adults were different from those reported for both U.S. Causcians and Europeans.

    Title Regulation of Activin's Access to the Cell: Why is Mother Nature Such a Control Freak?
    Date October 2000
    Journal Bioessays : News and Reviews in Molecular, Cellular and Developmental Biology
    Excerpt

    Activin A is a pluripotent growth factor with important roles in development, erythropoiesis and the local regulation of many tissues. At the post-translational level, the amount of activin A produced by cells may be modulated through the diversion of activin A subunits into the formation of inhibin or other activins containing heterodimeric forms. Once assembled, activin interacts with various low- and high-affinity binding proteins, such as follistatin and alpha(2)-macroglobulin, that have consequences for receptor availability. In common with other TGFbeta family members, activin signals through pairs of type I and II receptor kinases and the Smad intracellular signalling cascade. Other checkpoints have been identified such as the recently identified pseudoreceptor, BAMBI. These emerging findings point to a tightly coordinated regulation of the exposure of a cell or tissue to activin, consistent with the low amounts of this potent factor that are necessary to modulate cellular responses.

    Title Divergent Cell-specific Effects of Activin-a on Thymocyte Proliferation Stimulated by Phytohemagglutinin, and Interleukin 1beta or Interleukin 6 in Vitro.
    Date August 2000
    Journal Cytokine
    Excerpt

    Activin-A is a member of the transforming growth factor-beta (TGF-beta) cytokine family. Based on studies in several cell systems, activin-A has been postulated to be a specific inhibitor of the actions of the inflammatory cytokine, interleukin 6. In cultures of adult rat thymocytes, activin-A inhibited sub-optimal phytohemagglutinin-induced and interleukin 1beta-stimulated proliferation, as measured by [(3)H]-thymidine incorporation in vitro. In contrast with TGF-beta1, which exerted similar inhibitory effects on thymocyte proliferation, activin-A activity was reduced by increasing the concentration of phytohemagglutinin or addition of the reducing agent, beta-mercaptoethanol. Both activin-A and TGF-beta1 inhibited the in vitro production of interleukin 6 by thymocytes in the presence of phytohemagglutinin and interleukin 1beta. In the presence of exogenous interleukin 6, however, both activin-A and TGF-beta1 stimulated thymocyte proliferation. These data suggest that activin-A inhibits thymocyte growth and differentiation, at least in part, by inhibiting endogenous production of interleukin 6, but stimulates thymocyte growth when exogenous interleukin 6 is present in vitro. These data indicate that activin interacts with other cytokines to exert complex regulation of T cell development, and is not an inhibitor of interleukin 6 action in all cell systems.

    Title Release of Activin and Follistatin During Cardiovascular Procedures is Largely Due to Heparin Administration.
    Date August 2000
    Journal The Journal of Clinical Endocrinology and Metabolism
    Excerpt

    Recent evidence has suggested that activin A complexed to its binding protein, follistatin, may be present on the surface of cells through their interaction with heparan sulfate proteoglycans. As heparin is used routinely in many cardiovascular procedures for its anticoagulation properties, it may also cause the release of heparin-binding growth factors, including activin and follistatin, from the vascular endothelium. We examined the effect of two cardiovascular procedures and the use of heparin directly on the circulating concentrations of activin A and follistatin. A rapid and robust release of activin A and follistatin occurred in the circulation of patients undergoing abdominal aortic aneurysm repair or carotid endarterectomy at the time of vessel clamping and administration of heparin (5000 IU). This release pattern was dissimilar to that of the inflammatory marker, interleukin-1beta. However, administering heparin (2500 IU) to coronary angiography patients produced a similar activin and follistatin response, whereas placebo-treated angiography patients had no response. These findings illustrate that the routine use of heparin in surgical procedures elicits a rapid and robust release of activin and follistatin. This has direct clinical relevance by potentially activating heparin-binding growth factors that are important in injury, hyperplasia, and restenosis of vessels.

    Title Changes in Serum Inhibin, Activin and Follistatin Concentrations During Puberty in Girls.
    Date July 2000
    Journal Human Reproduction (oxford, England)
    Excerpt

    Serum concentrations of inhibin A, inhibin B, activin A and follistatin were determined using two-site enzyme-linked immunosorbent assays (ELISA) during pubertal ovarian development in 28 girls and five follicular phase women. Blood obtained every 15 to 20 min overnight was pooled for peptide determination. Serum inhibin A concentrations increased in mid puberty, exhibiting positive correlations with bone age (r = 0.527, P = 0.0016) and oestradiol concentrations (r = 0.581, P = 0.0005). Inhibin B concentrations peaked in mid puberty and declined thereafter, but remained greater than concentrations seen in prepubertal girls, and correlating positively with oestradiol (r = 0.362, P = 0.046) and follicle stimulating hormone (FSH) concentrations (r = 0.369, P = 0.038). Total activin A concentrations did not vary significantly across pubertal stages. Total follistatin concentrations, determined by radioimmunoassay, decreased with advancing puberty, exhibiting negative correlations with bone age (r = -0.634, P = 0.0001) and oestradiol concentration (r = -0.687, P = 0.0001). Follistatin concentrations determined by an ELISA specific for follistatin 288 were greatest in mid-pubertal girls, but concentrations in late puberty were less than those in early puberty. The free follistatin assay indicated that all circulating follistatin was activin-bound. These results suggest that significant changes in serum concentrations of FSH-regulatory peptides accompany the onset of puberty.

    Title The Roles of Inhibin and Related Peptides in Gonadal Function.
    Date July 2000
    Journal Molecular and Cellular Endocrinology
    Excerpt

    Inhibin A and B are dimeric proteins capable of suppressing FSH both in vitro and in vivo. The principal form in the male is inhibin B which is produced in the testis and circulates to inhibit pituitary FSH secretion. Activin A, B and AB are dimeric proteins that share the same beta subunits with the inhibins but, in contrast, stimulate FSH secretion. Although activin A circulates, castration does not lead to a decrease in serum concentrations, indicating that the testis is not the major source of activin A. In the circulation, the activins are bound to a structurally unrelated binding protein, follistatin, that neutralizes the biological actions of these proteins. The subunits of the inhibins/activins as well as follistatin are also produced locally within the pituitary and their levels can be modulated by testosterone and gonadotrophin releasing hormone as well as by autocrine mechanisms. Consequently, the output of FSH is dependent of the balance between local processes and the circulating feedback exerted by testosterone and inhibin. There is increasing data to support the local gonadal production of not only inhibin but also activin and follistatin by both germ cells and somatic cells such as the Sertoli cells. Evidence is accumulating to support the concept that these proteins exert local regulatory mechanisms in the testis.

    Title Activin A Release into the Circulation is an Early Event in Systemic Inflammation and Precedes the Release of Follistatin.
    Date May 2000
    Journal Endocrinology
    Excerpt

    Recent evidence suggests a role for activin A, and its binding protein, follistatin, in inflammatory pathways. However, whether activin is released systemically during inflammation is not known. In this study, a release of activin A into the circulation occurred in sheep within 1 hour of injection of lipopolysaccharide. This rapid peak in activin A preceded the release of the key inflammatory cytokines, tumor necrosis factor-alpha and interleukin-6. Follistatin release into the circulation occurred some 4 hours after the peak of activin A and continued out to 24 hours from lipopolysaccharide treatment. These data are the first to document a circulatory response of activin A to an inflammatory stimulus, and together with previous findings, suggest that activin A may have both pro- and anti-inflammatory actions in regulating cytokine-driven pathways.

    Title Bacterial Lipopolysaccharide-induced Inflammation Compromises Testicular Function at Multiple Levels in Vivo.
    Date January 2000
    Journal Endocrinology
    Excerpt

    While it is well known that serious illness and inflammation reduce male fertility, the mechanisms involved are poorly understood. In adult male rats, a single injection of lipopolysaccharide at doses that induced either mild or severe inflammation, caused a biphasic decline in Leydig cell testosterone production and gonadotropin responsiveness. In the high dose group only, serum LH levels also were reduced; however, intratesticular testosterone concentrations remained at a level adequate to support qualitatively normal spermatogenesis in both treatment groups. Testicular interstitial fluid formation also declined in a dose-dependent fashion after lipopolysaccharide treatment. In the high dose group only, these hormonal and vascular changes were accompanied by an increase in endothelial permeability, microhemorrhage, and inflammatory cells in the testis, followed by vacuolization of round spermatid nuclei, disruption of Sertoli-germ cell contacts at stages I-IV of the cycle of the seminiferous epithelium, and subsequently apoptosis of spermatocytes at stages II-V. These data indicate that mild inflammation causes local inhibition of Leydig cell function with relatively little spermatogenic damage. The pathological changes in spermatogenic function during severe inflammation are most likely due to direct effects of inflammatory mediators on the seminiferous epithelium or testicular vasculature, rather than inhibition of the brain-pituitary-Leydig cell axis.

    Title Uterine Milk Protein, a Novel Activin-binding Protein, is Present in Ovine Allantoic Fluid.
    Date October 1999
    Journal Endocrinology
    Excerpt

    Activins are pluripotent growth factors that have recently been shown to be present in placental and fetal membrane preparations. Our previous studies have identified and purified activin A from ovine amniotic and allantoic fluids. In this study, ligand blots of side fractions from the isolation of activin A from allantoic fluid suggested the presence of activin-binding proteins other than follistatin. Further purification of one of these fractions involved two sequential reverse phase HPLC steps and a Superose 12HR fractionation. SDS-PAGE revealed a single protein band of 55 kDa, which was identified by NH2-terminal sequencing as ovine uterine milk protein (UTMP), a member of the serine protease inhibitor (serpin) superfamily of proteins. Further binding studies, using ligand blot techniques and Superose 12HR fractionation in the presence of [125I]activin, demonstrated UTMP to be an activin-binding protein with a lower affinity for activin than that of follistatin. A study of the specific binding behavior of UTMP to activin, using surface plasmon resonance, revealed an apparent equilibrium dissociation constant (Kd) of 49 +/- 25 nM, compared with the follistatin-activin Kd of 379 +/- 51 pM. Similar to another activin-binding protein, alpha2-macroglobulin, UTMP was unable to neutralize the bioactivity of activin in a bioassay based on the capacity of activin to inhibit the proliferation of an MPC-11 plasmacytoma cell line. The high concentrations of this protein in uterine fluid during pregnancy and its ability to bind activin suggest that UTMP may act as a low affinity, high capacity binding protein to sequester activin in the local uterine environment.

    Title A Sensitive and Specific in Vitro Bioassay for Activin Using a Mouse Plasmacytoma Cell Line, Mpc-11.
    Date September 1999
    Journal The Journal of Endocrinology
    Excerpt

    A new in vitro bioassay for activin was developed using the mouse plasmacytoma cell line, MPC-11. Human recombinant (hr) activin A dose-dependently inhibited the proliferation of these cells, whereas a range of other factors, including inhibin, follistatin and transforming growth factor-beta1, -beta2 and -beta3 had no effect. Conditioned medium containing activin B induced an inhibition similar to hr-activin A. The inhibitory influence of activin A could be blocked by follistatin, but not by hr-inhibin A. This bioassay had a sensitivity for activin A of around 0.4 ng/ml, an ED50 response of 3.5 ng/ml, and an intra-assay coefficient of variation of <11%. It offers substantial advantages over existing in vitro activin bioassays in terms of ease of use, specificity and throughput. The utility of the MPC-11 bioassay was demonstrated in the purification of activin from amniotic fluid, where an almost identical profile of bioactive activin A was detected compared with the pituitary cell bioassay of activin. Bioactive activin could also be detected in unpurified ovine allantoic and amniotic fluids and bovine follicular fluid. Measuring activin in untreated and heat-treated human sera or seminal plasma was hampered by a non-specific inhibitory effect, so that several serum samples did not run parallel with the hr-activin A standard. This inhibitory effect by serum could not be overcome by addition of follistatin, suggesting it is not activin-like bioactivity. This new bioassay for activin demonstrates widespread applicability for monitoring of purified or partially purified samples during purification procedures, bioactivity measurements, receptor-binding studies and assays of cell culture medium.

    Title Activin A and Follistatin: Their Role in the Acute Phase Reaction and Inflammation.
    Date July 1999
    Journal The Journal of Endocrinology
    Title Activin A Regulates Growth and Acute Phase Proteins in the Human Liver Cell Line, Hepg2.
    Date June 1999
    Journal Molecular and Cellular Endocrinology
    Excerpt

    Activin, and its binding protein, follistatin, are up-regulated by mediators of inflammation, and recent studies have demonstrated that activin A can block the activity of the key inflammatory cytokine, interleukin-6 (IL-6). These findings thereby implicate activin and follistatin in the control of the inflammatory cascade. In this study, interactions between interleukin-1beta (IL-1beta), IL-6 and activin were examined the human liver cell line, HepG2, for their effect on cell proliferation and the production of the acute phase proteins, haptoglobin and alpha1-acid glycoprotein (alpha1-AGP). IL-1beta and activin A, but not IL-6, inhibited the proliferation of HepG2 cells. Activin A together with IL-1beta caused a greater inhibition of proliferation than either factor alone, and the inhibitory effects of activin A were blocked by the addition of follistatin to the cultures. Activin A alone inhibited the production of haptoglobin but did not affect alpha1-AGP concentrations. However, activin A suppressed the stimulatory effects of IL-6 on the production of both haptoglobin and alpha1-AGP. Production of follistatin by HepG2 cells was stimulated by activin A, but was inhibited by both IL-1beta and IL-6, indicating a complex regulatory loop is operable to modulate the effects of activin A during inflammation. Taken together, these data suggest that activin A interacts with IL-1beta and IL-6 to regulate and coordinate the production of acute phase proteins during an inflammatory episode.

    Title Deletion Polymorphism in the Angiotensin-converting Enzyme Gene As a Thrombophilic Risk Factor After Hip Arthroplasty.
    Date April 1999
    Journal Thrombosis and Haemostasis
    Excerpt

    Despite thromboprophylaxis, deep vein thrombosis is a common complication of major orthopedic surgery. Predisposing genetic risk factors are unknown. In this case-control study, we investigated the association of the insertion (I)/deletion (D) angiotensin converting enzyme (ACE) gene polymorphism, Factor V Leiden (R506Q) mutation, and 5,10 methylenetetrahydrofolate reductase (MTHFR) gene polymorphism with post-operative venous thrombosis in 85 patients who underwent elective total hip arthroplasty. The odds of a thrombotic event following hip surgery among subjects with the DD genotype of the ACE gene was increased more than 10-fold compared to subjects with the II genotype (odds ratio 11.7 [95% confidence interval 2.3-84.5]); it was increased 5-fold in subjects with the ID genotype compared to the II genotype (odds ratio 5.0 [95% confidence interval 1.1-34.9]). Mean plasma ACE level in control subjects not on ACE inhibitors at the time of study (n=43) was lowest in persons homozygous for the I allele (18.9+/-7.95 U/l), intermediate in patients with the ID genotype (31.6+/-10.8 U/l) and highest in subjects homozygous for the D allele (44.0+/-7.14 U/l). Mean plasma ACE level among cases was higher (33.0 U/l, n=25) than among controls (29.4 U/l, n=43) but this difference was not statistically significant. Neither the Factor V Leiden mutation nor MTHFR gene polymorphism increased the risk of thrombosis following hip replacement. These results demonstrate that the I/D ACE gene polymorphism is a potent risk factor for thrombosis in subjects undergoing total hip arthroplasty.

    Title Mechanisms of Protein Feedback on Gonadotropin Secretion.
    Date December 1998
    Journal Journal of Reproductive Immunology
    Excerpt

    The classical concept that the control of follicle-stimulating hormone (FSH) and luteinising hormone (LH) secretion by the pituitary gland is achieved by the stimulation of gonadotropin-releasing hormone (GnRH) and negative feedback by steroid secretion from the gonads has recently been modified by the identification of several proteins with the capacity to modulate FSH secretion. These proteins, inhibin, activin and follistatin, are produced in the ovary and testis and have the capacity to act as long loop protein feedback signals which modulate FSH secretion. Further studies indicate that inhibin probably acts as a circulating hormone to inhibit FSH secretion. More recent data suggest that activin and follistatin may act as local mediators to control FSH secretion at the pituitary by paracrine or autocrine phenomena. The production of activin, which stimulates FSH, and its binding protein follistatin, by pituitary cells raises the possibility that the local production modulates FSH secretion in addition to the long loop feedback signals emanating from the gonads through the steroid hormones estradiol and testosterone as well as circulating inhibin levels. Further studies are necessary to confirm the nature of these regulatory processes.

    Title Follistatin: a Multifunctional Regulatory Protein.
    Date December 1998
    Journal Frontiers in Neuroendocrinology
    Excerpt

    Follistatin was first described in 1987 as a follicle-stimulating hormone inhibiting substance present in ovarian follicular fluid. We now know that this effect of follistatin is only one of its many properties in a number of reproductive and nonreproductive systems. A majority of these functions are facilitated through the affinity of follistatin for activin, where activin's effects are neutralized through its binding to follistatin. As such, the interplay between follistatin and activin represents a powerful regulatory mechanism that impinges on a variety of cellular processes within the body. In this review we focus on the biochemical characteristics of follistatin and its interaction with activin and discuss the emerging role of these proteins as potent tissue regulators in the gonad, pituitary gland, pregnancy membranes, vasculature, and liver. Consideration is also given to the larger family of proteins that contain follistatin-like modules, in particular with regard to their functional and structural implications.

    Title Net Increase in Stimulatory Input Resulting from a Decrease in Inhibin B and an Increase in Activin A May Contribute in Part to the Rise in Follicular Phase Follicle-stimulating Hormone of Aging Cycling Women.
    Date October 1998
    Journal The Journal of Clinical Endocrinology and Metabolism
    Excerpt

    Recent studies suggest that an age-related decline in ovarian inhibin B may play a role in the increase in follicular phase FSH in menstrual cycles of older women. Considering that the peripheral feedback regulation of FSH is dictated by the overall tone of inhibins, activins, and follistatins as well as estradiol, it is essential to determine the relative inputs of all of these regulators in assessing whether the collective peripheral input to FSH is one of inhibition or stimulation. To test the hypothesis that changes in the overall tone of peripheral feedback may contribute to this hallmark sign of aging, we compared the concentrations of dimeric inhibin A, inhibin B, activin A, and total and free follistatin in 7 young (mean age, 27.9 +/- 2.6 yr) and 10 older (mean age, 43.6 +/- 0.9 yr) cycling women during the follicular (FOLL; cycle day 6) and midluteal (ML; 7 days post-LH surge) phases of the menstrual cycle. Subjects were preselected on the basis of FOLL phase FSH levels (older, > or = 8.0 mIU/mL; younger, < 8 mIU/mL). Circulating FSH regulatory peptide concentrations were determined from samples pooled from blood drawn every 10 min for 8 daytime h using specific 2-site assays. In the older group, cycle length was shorter (29.1 +/- 0.5 vs. 26.1 +/- 0.5, young vs. older; P < 0.001), mean LH levels during the follicular phase were higher (LH, 5.6 +/- 0.8 vs. 8.8 +/- 1.1 mIU/mL, young vs. older; P < 0.001). Mean FSH levels for the older and younger groups averaged 10.8 +/- 0.8 and 6.2 +/- 0.3 mIU/mL, respectively. Estradiol levels were higher, but not statistically different, than those in the younger group (99 +/- 13 vs. 169 +/- 25 pmol/L, young vs. older; P = 0.06). In both age groups, inhibin B levels were higher in the FOLL vs. ML phase, inhibin A levels were higher in the ML vs. FOLL phase, but total activin A and total and free follistatin did not differ across cycle days. FOLL phase inhibin A levels were higher in the older group (16.3 +/- 2.4 vs. 26.4 +/- 3.4 pg/mL, young vs. older; P = 0.024), but levels of inhibin B were lower (323 +/- 80 vs. 163 +/- 24 pg/mL, young vs. older; P = 0.03). Overall, the estimated total inhibin activity (inhibin A plus inhibin B) was lower in older cycling than in younger women (339 +/- 82 and 189 +/- 24 pg/mL, young vs. older). Total and free follistatin levels were not different among the 2 groups of women. In contrast, total activin A levels were higher in the older cycling group (0.51 +/- 0.05 and 0.68 +/- 0.05 ng/mL, young vs. older; P = 0.02). No differences in age groups were observed during the ML phase for any of the variables measured. These data suggest that a net increase in stimulatory input resulting from a decrease in inhibin B and an increase in activin A may contribute in part to the monotropic FSH increase in aging women.

    Title The Up-regulation of Il-6 and Il-8 in Human Endothelial Cells by Activated Protein C.
    Date September 1998
    Journal Journal of Immunology (baltimore, Md. : 1950)
    Excerpt

    The protein C/protein S anticoagulant pathway has been proposed to be a common link between coagulation and inflammation. Studies have suggested that a component of the anticoagulant pathway, activated protein C (APC), may play a role in the inflammatory response by modulating the effects of cytokines such as TNF and by blocking neutrophil activation. Cytokines are known to be intimately involved in the inflammatory response and to function in part to restore hemostatic balance. To begin to delineate what role APC may have in the inflammatory response, we have investigated the effect of APC on the production of the proinflammatory cytokines IL-6 and IL-8 in primary HUVEC, human microvascular endothelial cells, and human coronary artery endothelial cells. Our results have demonstrated that physiologic concentrations of APC significantly up-regulated the production of both IL-6 and IL-8. This increase, which was seen at both the RNA and protein level, was not due to either thrombin or LPS contamination of the APC preparation. Additional studies also showed that the APC-mediated up-regulation of IL-6 and IL-8 was IL-1 independent. Although neither purified protein C nor protein S alone had an effect on cytokine production, protein S, the cofactor for APC, significantly enhanced the ability of APC to up-regulate IL-6/IL-8 production. These results provide further evidence for a role for APC in the inflammatory response.

    Title Localization of Follistatin in the Rat Testis.
    Date July 1998
    Journal Journal of Reproduction and Fertility
    Excerpt

    The cellular localization of the activin-binding protein, follistatin, in the rat testis has been a matter of some controversy with different investigators claiming that Sertoli cells, Leydig cells or germ cells are the primary cell types containing this protein. The localization of mRNA encoding follistatin was re-examined using reverse transcription-polymerase chain reaction (RT-PCR) and in situ hybridization as well as the distribution of follistatin by immunohistochemistry. The results demonstrate that mRNA encoding follistatin is located in many germ cells including type B spermatogonia, primary spermatocytes with the exception of the late leptotene and early zygotene stages, and spermatids at steps 1 to 11. It is also found in Sertoli cells and endothelial cells but not in Leydig cells. Immunohistochemistry, using two different antisera to follistatin, showed that this protein was localized to spermatogonia, primary spermatocytes at all stages except the zygotene stage, spermatids at all stages and to endothelial cells and Leydig cells in the intratubular regions. The failure to detect mRNA for follistatin in Leydig cells using RT-PCR and in situ hybridization suggests that the immunohistochemical localization in these cells reflects binding of follistatin produced elsewhere. The widespread localization of follistatin, taken together with its capacity to neutralize the actions of activin, may indicate that follistatin modulates a range of testicular actions of activin, many of which remain unknown.

    Title Guide to Evaluation of Permanent Impairment of the Temporomandibular Joint. American Academy of Head, Neck and Facial Pain; American Academy of Orofacial Pain; American Academy of Pain Management; American College of Prosthodontists; American Equilibration Society and Society of Occlusal Studies; American Society of Maxillofacial Surgeons; American Society of Temporomandibular Joint Surgeons; International College of Cranio-mandibular Orthopedics; Society for Occlusal Studies.
    Date May 1998
    Journal Cranio : the Journal of Craniomandibular Practice
    Title Follistatin Has a Biphasic Response but Follicle-stimulating Hormone is Unchanged During an Inflammatory Episode in Growing Lambs.
    Date March 1998
    Journal The Journal of Endocrinology
    Excerpt

    The effects on plasma follistatin concentrations of an inflammatory episode, induced by the intrathoracic injection of yeast, were examined in growing lambs; this model results in acute loss of appetite, food intake and liveweight and the activation of the acute-phase pathway for several weeks as adjudged by the production of haptoglobin and other acute-phase proteins. In these animals (n = 8) there was a biphasic response in follistatin concentrations, with an initial 200% increase (P < 0.001) in follistatin within 24 h of injection of yeast. Thereafter, follistatin concentrations were depressed to 70% of pretreatment levels 48 h after injection (P < 0.01), followed by a gradual recovery of concentrations to pretreatment values. In another group of lambs (n = 16) that were feed-restricted to mimic the reduced food intakes and liveweight changes in the yeast-injected group, plasma follistatin was also reduced to around 70% of pretreatment levels (P < 0.01) within 1 day of the dietary regimen being implemented, followed by a gradual return to pretreatment values as food intakes were increased. Plasma follistatin correlated significantly (r = 0.57, P < 0.0001) with food intake, but not with liveweight changes. Plasma follistatin concentrations were unchanged in a third group fed ad libitum (n = 8), except during two periods when food intakes were significantly (P < 0.05) reduced, when follistatin concentrations also decreased (P < 0.01). Plasma follicle-stimulating hormone (FSH) concentrations in the three groups of lambs were not significantly affected by the treatment regimes or changes in follistatin concentrations. These findings indicate that peripheral follistatin concentrations are modulated by both inflammatory and nutritional mechanisms, and that significant fluctuations in follistatin levels can occur without detectable perturbations in FSH secretion.

    Title Inhibin, Activin and Follistatin Bind Preferentially to the Transformed Species of Alpha 2-macroglobulin.
    Date December 1997
    Journal The Journal of Endocrinology
    Excerpt

    alpha 2-Macroglobulin (alpha 2-M), a major serum glycoprotein, has been implicated as a low-affinity binding protein for inhibin and activin. In serum, alpha 2-M exists as two major species, a native form that is abundant and stable, and a transformed ('fast') species that is rapidly cleared from the circulation via alpha 2-M receptors. In this study inhibin, activin and their major binding protein follistatin were investigated for their ability to bind to the native or transformed species of purified human alpha 2-M. Using native PAGE and size exclusion chromatography, radiolabelled inhibin, activin and follistatin bound to the transformed alpha 2-M. Inhibin and follistatin did not bind significantly to native alpha 2-M, whereas activin was able to bind to the native species but with a lower capacity compared with that to transformed alpha 2-M. Under reducing conditions, binding of these hormones to alpha 2-M was abolished. These findings implicate alpha 2-M as a mechanism whereby inhibin, activin and follistatin may be removed from the circulation through alpha 2-M receptors, but also whereby activin can be maintained in the circulation through its ability to bind to native alpha 2-M.

    Title Changes in the Isoforms of Luteinizing Hormone and Follicle-stimulating Hormone During Puberty in Normal Children.
    Date October 1997
    Journal The Journal of Clinical Endocrinology and Metabolism
    Excerpt

    Concentrations of LH and FSH are known to increase during normal pubertal development, but changes in the isoforms of the gonadotropins at this time have not been investigated in depth. We examined the median charge of serum LH and FSH using agarose suspension electrophoresis in 81 normal children at pubertal stages I-V. In pubertal girls there were no significant (P > 0.05) differences in the median charge of LH, but there was a small (P = 0.05) shift to more acidic FSH isoforms between pubertal stages I and IV. In boys there was a significant (P < 0.01) shift to more acidic isoforms for both LH and FSH by pubertal stage II. Further changes were not found later in puberty. Except for LH at pubertal stage I, where the median charge was similar (P > 0.05) for both sexes, the median charge was more basic (P < 0.001) for both LH and FSH in girls compared with boys at all five pubertal stages. The degree of charge heterogeneity of FSH, estimated as the peak width at half the peak height, was significantly (P < 0.01) larger at pubertal stage I than at pubertal stages III-V in both boys and girls. The charge heterogeneity of LH was similar for all pubertal stages in both sexes. In conclusion, there were few qualitative changes in the gonadotropins during normal female puberty, whereas in the male there was a dramatic shift to more acidic isoforms of LH and FSH early in puberty. This information may assist our understanding of normal and pathological processes during puberty and may be of clinical relevance in detecting the initiation of puberty in boys.

    Title A New Small Particle Packing for Faster Analysis with High Resolution.
    Date September 1997
    Journal Journal of Pharmaceutical and Biomedical Analysis
    Excerpt

    The need for fast and efficient separations of complex samples such as pharmaceuticals and biologicals led to the development of fast, efficient, and reproducible 3.5 microns columns. Separations using 3.5 microns column are 30-50% faster at equal plate-count compared to 5 microns columns. The results show that the 3.5 microns columns (100 mm length) give the same efficiency and resolution of drug impurities as the 5 microns columns (150 mm length). For many analytical methods, switching to 3.5 microns columns saves time and reduces costs. Separation methodologies using 5 microns columns are easily modified to accommodate 3.5 microns columns of the same chemistry because efficiency, resolution and sensitivity remain the same. It is shown that 3.5 microns columns have lifetimes comparable to 5 microns columns of the same brand.

    Title Endothelial Cell Protein S Synthesis is Upregulated by the Complex of Il-6 and Soluble Il-6 Receptor.
    Date August 1997
    Journal Thrombosis and Haemostasis
    Excerpt

    We have recently demonstrated that the proinflammatory cytokine, interleukin-6 (IL-6), could upregulate the production of protein S in the human hepatoma cell line, HepG-2, but not in endothelial cells. In this study, we have demonstrated that the combination of exogenous IL-6 and soluble IL-6 receptor (sIL-6R) could significantly upregulate protein S production in both primary human umbilical vein endothelial cells (HUVEC) and in the immortalized human microvascular endothelial cell line, HMEC-1. The IL-6/sIL-6R complex was also able to rapidly induce tyrosine phosphorylation of the IL-6 transducer, gp130. Neutralizing antibodies directed against either IL-6 or gp130 blocked protein S upregulation by the IL-6/sIL-6R complex. It was also observed that exogenous sIL-6R could also upregulate protein S by forming a complex with IL-6 constitutively produced by the endothelial cell. Two other cytokines which also utilize the gp130 receptor, oncostatin M (OSM) and leukemia inhibitory factor (LIF), were also able to upregulate endothelial cell protein S. This study demonstrates a mechanism that allows endothelial cells to respond to IL-6 and also illustrates the potential importance of circulating soluble receptors in the regulation of the anticoagulation pathway.

    Title Ultrasonic Color Flow Mapping: the Visualization of Four-dimensional Cardiac and Vascular Flow Phenomena Using Two Dimensions and "real Time".
    Date July 1997
    Journal Ultrasound in Medicine & Biology
    Title Effects of the Booroola Gene (fecb(b)) on Bodymass, Testis Development and Hormone Concentrations During Fetal Life.
    Date March 1997
    Journal Journal of Reproduction and Fertility
    Excerpt

    In female fetuses the Booroola gene (FecB(B)) is known to affect germ cell development and consequently the pattern of ovarian follicular growth during fetal and post-natal life. However in males, the role of this gene during fetal development is unknown. The aims of the study reported here were to examine the effects of the FecB(B) gene on development of male fetuses with respect to body and organ mass for example, pituitary gland, adrenal and mesonephros), testes development, including numbers of germ cells, and also the plasma concentrations or tissue contents of the reproductive hormones (FSH, LH and testosterone) at days 40, 55, 75, 90 and 135 of gestation. The FecB(B) gene was found to influence litter size, bodymass, crown-rump length and testis mass at most stages of gestation. Some effects were also noted on the mesonephros at days 40 and 55 and on the pituitary and adrenal at days 90 or 135 of gestation. However, the FecB(B) gene was not observed to have an effect on the patterns of germ cell development, on pituitary content or plasma concentrations of immunoreactive or bioactive FSH or immunoreactive LH or testicular content of testosterone. When embryo transfer experiments were performed to eliminate the effects of litter size at days 40, 90 and 135 of gestation nearly all of the differences in bodymass, crown-rump length and organ mass disappeared. The only exception to this was at day 90 when bodymass continued to be lighter and crown-rump lengths smaller in the BB/B + fetuses compared with the +2 fetuses; the significance of this finding remains unknown. It is concluded that for Booroola male fetuses there are no direct effects of the FecB(B) gene on pituitary gonadotrophin function or testicular development after sexual differentiation. Moreover, although there may be temporal differences around day 90 of gestation, there are no long-term, direct effects of the FecB(B) gene on total body, adrenal, testis or pituitary mass. Collectively these findings for the male are similar to those for female fetuses except with regard to germ cell development.

    Title Oligonucleotide Ligation Assay for Detection of the Factor V Mutation (arg506-->gln) Causing Protein C Resistance.
    Date January 1997
    Journal Thrombosis Research
    Excerpt

    A point mutation in the Factor V (FV) gene at the activated protein C cleavage site, FV Arg (R)506-->FV Gln (Q)506, is the reported molecular basis for resistance to activated protein C (APC-R). This mutation has been reported in approximately 20-50% of individuals with previously unexplained thrombophilia and 3-5% of the general population. We have adapted an oligonucleotide ligation assay (OLA) for nonisotopic detection of the FV:Q506 mutation which permits rapid screening for this mutation. First, the polymerase chain reaction (PCR) was used for target DNA amplification, thus permitting nonisotopic reporters in the DNA analysis. Then thermostable ligase was used for ligation or covalent coupling of adjacent wild-type and mutant oligonucleotide probes which occurs only when the probes are annealed to a matched amplicon. A colorimetric ELISA-based detection assay was then used to capture 5' biotinylated probes in 96-well streptavidin-coated plates and by virtue of ligation, detection of a 3' digoxigenin reporter probe. Following the addition of anti-digoxigenin conjugate and enhanced alkaline phosphatase signal amplification, colorimetric substrate change was measured in an ELISA plate reader. This assay correctly identified FV genotypes of 290 samples.

    Title Successful Permanent Impairment Conference Catalyzes Unity.
    Date December 1996
    Journal Cranio : the Journal of Craniomandibular Practice
    Title Follistatin Concentrations in Male Sheep Increase Following Sham Castration/castration or Injection of Interleukin-1 Beta.
    Date December 1996
    Journal The Journal of Endocrinology
    Excerpt

    Plasma follistatin (FS) concentrations were determined after castration (n = 5) or sham castration (n = 4) of mature rams. Both treatments resulted in a prolonged increase in FS between 7 and 19 h after surgery, which returned to pretreatment concentrations by 24 h. Tumour necrosis factor-alpha (TNF-alpha), a sensitive maker of an acute-phase response, was undetectable in plasma, indicating that the FS response was not induced by trauma due to surgery. In a second experiment, injection of castrated rams (n = 4) with ovine recombinant interleukin-1 beta, an acute-phase mediator, resulted in a sustained rise in FS concentrations within 4 h of injection. Plasma TNF-alpha concentrations increased transiently within 1 h of interleukin-1 beta injection, indicating that an acute-phase response had been initiated. Plasma follicle-stimulating hormone (FSH) concentrations were significantly decreased at 8 and 24 h after interleukin-1 beta injection, strongly suggestive of an inhibitory effect of increased FS concentrations on the secretion of FSH. Injection of castrated rams (n = 2) with a control preparation of recombinant interleukin-2 did not induce an acute-phase response, and plasma FS and FSH concentrations were unaffected. These data show that the testis is not a major source of circulating FS, that the increase in circulating FS following sham castration/castration is not due to an acute-phase response, but that conversely FS concentrations are modulated by the acute-phase mediator, interleukin-1 beta.

    Title Tnf-alpha Suppresses Il-1 Alpha and Il-6 Upregulation of C4b-binding Protein in Hepg-2 Hepatoma Cells.
    Date October 1996
    Journal Thrombosis Research
    Excerpt

    C4b-binding protein (C4BP) is a regulatory protein involved in the regulation of the classical pathway of complement and the natural anticoagulant pathway. C4BP is synthesized by the liver, a target organ of the IL-6 proinflammatory cytokine. C4BP is an acute-phase protein and its basal levels may increase by as much as 4-fold during an inflammatory response. IL-6 which plays a major role in the modulation of the acute-phase proteins, including C4BP, also has been shown by our group to significantly increase hepatocyte production of the anticoagulant protein, protein S. In this study, we have examined the role of cytokine combinations on the production of the C4BP regulatory protein in the HepG-2 hepatoma cell line and report that IL-1 alpha and IL-6 in combination as well as IL-1 alpha and Oncostatin M (OSM) were approximately additive in the upregulation of C4BP while IL-6 and OSM were not and that TNF-alpha blocked both IL-1 alpha and IL-6 but not OSM upregulation of C4BP.

    Title Tnf-alpha Suppresses Il-6 Upregulation of Protein S in Hepg-2 Hepatoma Cells.
    Date October 1996
    Journal Thrombosis Research
    Excerpt

    The pathogenesis of disseminated intravascular coagulation (DIC) has, in part, been attributed to the impairment of the natural anticoagulant protein C/protein S pathway. DIC, which frequently occurs during sepsis, has been linked to cytokines that can induce or modulate procoagulant activity. Three of these cytokines, IL-1 alpha, IL-6, and TNF-alpha have been reported to be increased in the early stages of sepsis. In the present study, we have stimulated HepG-2 hepatoma cell cultures with recombinant human IL-1 alpha, IL-6, TNF-alpha, and oncostatin M (OSM). The results demonstrated that TNF-alpha, and to a lesser degree, IL-1 alpha, could significantly suppress IL-6 upregulation of protein S, whereas the effects of OSM was only suppressed by the combination of IL-1 alpha and TNF-alpha. The combination of IL-1 alpha and TNF-alpha also suppressed protein S production below that of control or basal levels. These results indicate that IL-1 alpha and TNF-alpha may play important regulatory roles in coagulation.

    Title Short-term Regulation of Gonadotropin Subunit Mrna Levels by Estrogen: Studies in the Hypothalamo-pituitary Intact and Hypothalamo-pituitary Disconnected Ewe.
    Date August 1996
    Journal Journal of Neuroendocrinology
    Excerpt

    In this study the levels of mRNA for the pituitary gonadotropin hormone subunits luteinizing hormone beta (LHbeta), follicle stimulating hormone beta (FSHbeta) and the common alpha-subunit were assessed during the acute feedback stages of estradiol benzoate (EB) action in ovariectomized (OVX) ewes with and without hypothalamo-pituitary disconnection (HPD). In OVX/HPD ewes maintained on hourly pulses of 250 micrograms of gonadotropin-releasing hormone (GnRH) a single i.m. injection of EB in oil caused a biphasic (decrease and then increase) change in plasma LH levels and a monophasic decrease in FSH levels. There was a decrease in pituitary alpha-subunit and FSHbeta mRNA levels during the acute negative (8 h post EB) and through the positive feedback (20 h post EB) stages of the response. No significant change was seen in LHbeta mRNA levels following treatment with EB. In hypothalamic-pituitary intact OVX ewes the same EB treatment as above caused a biphasic change in LH secretion with the positive feedback component being much greater than in GnRH-pulsed OVX-HPD ewes. The levels of mRNA for all three gonadotropin subunits were reduced by 8 h after EB injections and remained low throughout the positive feedback period. These data suggest that the LH surge in this experimental model does not require an increase in LHB mRNA levels. Furthermore, the fall in LHbeta subunit mRNA seen after estrogen injection of OVX ewes is most likely due to an effect of estrogen to decrease GnRH secretion, since pulsatile GnRH replacement prevents this effect. These data also show that estrogen feedback can effect rapid alterations in pituitary gonadotropin subunit mRNA levels. Short-term changes in FSHbeta mRNA are reflected in changes in FSH secretion; the same is not true for LH.

    Title Median Charge of Gonadotrophin Isoforms in the Pituitary Gland and in the Circulation of Sheep Fetuses from Mid- to Late Gestation.
    Date August 1996
    Journal The Journal of Endocrinology
    Excerpt

    The heterogeneity of LH and FSH in the sheep fetus was studied by determining the median charge of pituitary and circulating isoforms. Pituitary extracts from male and female fetuses at days 75, 95, 120 and 135 of gestation were subjected to agarose suspension electrophoresis. For all fetuses except the day 75 age group, the median mobility of the gonadotrophin isoforms in matching serum samples from the individual fetuses were also determined. LH and FSH in extracts, peripheral samples and column eluates were measured using sensitive and specific sandwich fluoroimmunoassays for ovine gonadotrophins. The median charge of pituitary LH became more basic (P < 0.001) with gestational age, whereas for pituitary FSH more acidic forms (P < 0.001) were present in the older groups. The female fetuses had more basic pituitary isoforms of LH than the males (P < 0.01) between days 95 and 135, and for FSH at day 75 (P < 0.05). In the matching serum samples, the median charge of the LH (P < 0.001) and FSH (P < 0.05) isoforms were more acidic than those in the pituitary gland. No significant effects of age or sex were detected in the median charge of the gonadotrophin isoforms in serum, but in a number of instances the median charge could not be determined due to low serum concentrations which affected the group sizes. These data show that in the sheep fetus LH and FSH are differentially regulated in qualitative as well as quantitative terms, and that the charge of fetal gonadotrophin isoforms changes according to the age and sex of the fetus.

    Title Orthopedic Manifestations of Acute Pediatric Leukemia.
    Date July 1996
    Journal The Orthopedic Clinics of North America
    Excerpt

    The variety and distribution of skeletal lesions in children with acute lymphoblastic leukemia is rarely seen in other diseases. Skeletal radiographic changes that can occur in a child with acute leukemia include diffuse osteopenia, metaphyseal bands, periosteal new bone formation, geographic osteolysis, osteosclerosis, mixed osteolysis and sclerosis, and permeative destruction. It is important for orthopedic surgeons to recognize the skeletal manifestations of acute leukemia of childhood because the physician who initially evaluates the child will often be an orthopedic surgeon, and a delay in diagnosis has an adverse affect on survival.

    Title More Basic Isoforms of Serum Gonadotropins During Gonadotropin-releasing Hormone Agonist Therapy in Pubertal Children.
    Date February 1996
    Journal The Journal of Clinical Endocrinology and Metabolism
    Excerpt

    An acute challenge of exogenous GnRH elicits rapidly increased serum gonadotropin levels with qualitative changes to more basic isoforms of both FSH and LH. Chronic GnRH agonist therapy suppresses endogenous gonadotropins, and the serum levels of FSH and LH are low and fairly constant. A possible qualitative change in the gonadotropins during GnRH agonist therapy was investigated by determination of the median charge of the gonadotropin isoforms before and during therapy in 18 pubertal children. Two different GnRH agonists were studied: buserelin, given intranasally or as a sc implant for 1.5-34 months to five girls, aged 7-10 yr, and for 5-6 months to two boys, aged 11-13 yr; and triptorelin, administered as a depot preparation for 3-6 months to four girls, aged 9-12.5 yr, and for 1-24 months to seven boys, aged 10.5-12 yr. FSH and LH in serum and eluates after electrophoresis in 0.10% agarose suspension were measured with sandwich fluoroimmunoassays. The mean serum FSH and LH levels decreased significantly (P < 0.05) in girls during triptorelin therapy, whereas only the FSH level decreased (P < 0.05) in the boys. There were no significant (P > 0.05) changes in serum gonadotropin levels during buserelin therapy. All of the children had more basic serum isoforms of LH, and all but one had more basic forms of FSH during the GnRH agonist treatments. In a girl who had more basic gonadotropin isoforms after treatment with triptorelin for 2 and 6 months, a GnRH challenge elicited the release of still more basic isoforms. The changes in mean median charge to more basic gonadotropin isoforms were highly significant for both busereline (P < 0.01) and triptorelin (P < 0.001) treatment. An increased (P < 0.001) degree of charge heterogeneity was observed for FSH after triptorelin therapy. These findings show that there is a qualitative change in the isoforms of both FSH and LH in serum during GnRH agonist therapy in pubertal children. The changes in charge to more basic gonadotropin isoforms most likely reflect a direct effect at the pituitary level, leading to the synthesis and/or selective release of less sialylated and sulfated isoforms of the gonadotropins. The observed qualitative changes in the gonadotropin isoforms in these pubertal children may be part of the clinical effects of GnRH agonist therapy, leading to an arrest or regression of puberty.

    Title Il-6 Upregulates Protein S Expression in the Hepg-2 Hepatoma Cells.
    Date December 1995
    Journal Thrombosis and Haemostasis
    Excerpt

    Several pro-inflammatory cytokines have been shown to be important in the modulation of the procoagulant response. However, what role these cytokines may have in the regulation of coagulation inhibitors is poorly understood. While the hepatocyte is a primary site of synthesis for the anticoagulant protein C and S, it is also a major target cell for the proinflammatory cytokine, IL-6. We have found that stimulation of HepG-2 hepatoma cells with IL-6 (5 ng/ ml) significantly increased the production of the anticoagulant cofactor, protein S, in both a time and dose dependent fashion. This increase was seen at both the RNA and protein level. A mouse monoclonal neutralizing antibody to human IL-6 suppressed the IL-6 effect in a concentration dependent fashion. IL-6 also increased the release of the C4b-binding protein but had no effect on protein C production. When combined with either dexamethasone or soluble IL-6 receptor, the IL-6 response was significantly enhanced. Oncostatin M, a functionally related cytokine, had a similar effect while other related cytokines, IL-11 and leukemia inhibitory factor, only had a marginal effect. IL-1, TGF-beta, and TNF-alpha had no significant effect on protein S production. These results indicate that IL-6 may play an important regulatory role in the anti-coagulant pathway.

    Title Hemostatic Properties of the Sv-40 Transfected Human Microvascular Endothelial Cell Line (hmec-1). A Representative in Vitro Model for Microvascular Endothelium.
    Date October 1995
    Journal Thrombosis Research
    Excerpt

    HMEC-1 is a SV-40T transfected human microvascular endothelial cell line that constitutively expresses RNA transcripts for plasminogen activator inhibitor 1 (PAI-1), tissue-type plasminogen activator (t-PA), protein S (PS), von Willebrand factor (vWF), and thrombomodulin. Tissue factor (TF) can be induced in response to stimulation with tumor necrosis factor alpha (TNF-alpha), interleukin-1 alpha (IL-1alpha) and phorbol 12-myristate 13-acetate (PMA). Proteins corresponding to PAI-1, t-PA, protein S and vWF genes were constitutively released in the culture supernatant. This cell line is a model that will be useful to investigate coagulation/fibrinolytic properties of microvascular endothelium.

    Title Effects of Ovariectomy and Fecbb Genotype on the Median Charge and Circulating Half-life of Pituitary Fsh Isoforms of Ewes.
    Date September 1995
    Journal Journal of Reproduction and Fertility
    Excerpt

    This study investigated the effects of short-term (20 days) ovariectomy, the effects of FSH assay (radioimmunoassay, receptor assay or in vitro bioassay) and of FecBB genotype on the characteristics of pituitary FSH from Booroola ewes. Pituitary extracts were obtained from ovariectomized homozygous carriers (BB) and non-carriers (++, n = 8 per genotype) and ovary-intact controls (n = 4 per genotype). The extracts (n = 4 per genotype per treatment) were subjected to agarose suspension electrophoresis and the eluates were assayed by the three FSH methods. There were significant effects of ovariectomy (P < 0.01) and assay system (P < 0.05) but not of genotype on the median charge of FSH isoforms. The mean +/- SEM migration rates for FSH in intact and ovariectomized ewes were 0.469 +/- 0.006 and 0.439 +/- 0.006 albumin mobility units, respectively (P < 0.01), indicating a shift to more basic isoforms after short-term ovariectomy. When the pituitary extracts were subjected to anion-exchange HPLC, there was a significant (P < 0.01) shift to more basic isoforms in the ovariectomized ewes as shown using agarose electrophoresis, and no gene effects were noted. When the pituitary extracts (n = 4-8 per group) were injected into mature female mice, there were no significant effects of ovariectomy or genotype on the circulating half-lives of the pituitary FSH isoforms. These results indicate that after short-term ovariectomy, the increase in plasma FSH concentrations is accompanied by a shift in the median charge of pituitary FSH isoforms without any observable change in their metabolic clearance rates.(ABSTRACT TRUNCATED AT 250 WORDS)

    Title Hypertensive Encephalopathy Secondary to a Phaeochromocytoma: the Cause of Death After Carotid Endarterectomy. Case Report.
    Date May 1995
    Journal The Journal of Cardiovascular Surgery
    Excerpt

    A female patient who underwent a carotid endarterectomy, died following a hypertensive crisis which may have been due to an unknown phaeochromocytoma.

    Title Bioactive Follicle-stimulating Hormone Concentrations in Plasma During the Estrous Cycle of the Ewe.
    Date April 1995
    Journal Biology of Reproduction
    Excerpt

    Bioactive FSH (B-FSH) concentrations in plasma were determined during the ovine estrous cycle by means of an in vitro bioassay. The concentrations of B-FSH were elevated during and after the preovulatory LH surge and were significantly (p < 0.05) lower during the late-luteal to mid-follicular phases compared with the mid-luteal phase. A significant (p < 0.05) increase in B-FSH was found about 36 h before the LH surge, at a time when the immunoreactive FSH (I-FSH) concentrations were low and unchanged. The plasma B/I ratio for FSH was relatively constant during the luteal phase; it then increased significantly (p < 0.05) before the LH surge and decreased again at the time of the LH surge itself. Pulsar analysis showed that there were 4 peaks of B-FSH throughout the estrous cycle with 2 during the luteal phase, 1 after the LH surge, and the other either during the follicular phase or associated with the LH surge. For I-FSH there were approximately 8 peaks throughout the cycle with 4 during the luteal phase, 2 after the LH surge, and 1 each during the follicular phase and the preovulatory LH surge. There was a weak negative correlation between I-FSH and immunoreactive inhibin (I-inhibin) during most of the estrous cycle, but B-FSH and the B/I ratio were only correlated (negatively) with I-inhibin in the 24 h before the preovulatory LH surge. These findings suggest that there are significant changes in the circulating isoforms of FSH during the ovine estrous cycle that may affect the growth of antral follicles developing towards ovulation.

    Title Effect of Chronic Daily Oral Administration of 17 Beta-oestradiol and Norethisterone on the Isoforms of Serum Gonadotrophins in Post-menopausal Women.
    Date April 1995
    Journal Clinical Endocrinology
    Excerpt

    Chronic treatment with 17 beta-oestradiol (E2) implants has been found to counteract the formation of more acidic isoforms of the gonadotrophins in post-menopausal women. Oral medication with an oestrogen in combination with a progestagen is a common hormone replacement therapy (HRT) in post-menopausal women. The present study investigated the effect of such a therapy on the concentration and charge of the gonadotrophin isoforms in serum.

    Title Femur Fractures with Femoral or Popliteal Artery Injuries in Blunt Trauma.
    Date March 1995
    Journal Journal of Orthopaedic Trauma
    Excerpt

    The treatment and results of 13 blunt femoral fractures with an arterial injury were reviewed. Two of the 13 patients (15%) sustained segmental (two levels) arterial injuries. Stabilization of the femur fractures were performed before arterial repair in 10 of the 13 femurs. The results were determined at an average of 4.5 years. For the eight open fractures, two patients had above-knee amputations, no limb regained > 90 degrees of knee motion, four patients required a brace or cane, and three patients have intermittent wound drainage. The five closed fractures all regained full function with full knee motion. Due to the 15% incidence of segmental arterial injury, "wide-field" arteriography is advised for the evaluation of this injury. Femoral stabilization may be performed before arterial repair if the procedure does not encroach on the viability of the limb. The functional results depend on the soft-tissue damage, as illustrated by the poor results seen in open fractures.

    Title Development of an Elisa for Quantification of Human Protein S in Cell Culture Fluids Using Commercial Polyclonal Antisera.
    Date February 1995
    Journal Journal of Immunoassay
    Excerpt

    An enzyme-linked immunosorbent assay (ELISA) was developed to measure protein S antigen released into cell culture fluids. We used readily available commercial polyclonal antisera to develop the assay. This assay was sensitive with a detection limit of about 0.086 ng/ml. Between-assay precision (coefficient of variation) at levels of 0.2, 1.1, and 13.9 ng/ml was 14%, 15%, and 11% respectively. Specificity and accuracy were demonstrated from the use of: 1) culture fluids from 3-primary endothelial cell cultures and 7-cell lines known to constitutively produce protein S; 2) 2-cell lines not synthesizing protein S; and 3) from selected samples of normal and protein S deficient plasma. The ELISA described here was about 12-fold more sensitive and 40-fold more cost effective when compared to a commercial ELISA kit. Thus the assay provided a sensitive, specific, precise and economical method useful for the measurement of the nanogram amounts of protein S commonly encountered in cell culture fluids.

    Title Serum Gonadotropin Isoforms Become More Basic After an Exogenous Challenge of Gonadotropin-releasing Hormone in Children Undergoing Pubertal Development.
    Date October 1994
    Journal The Journal of Clinical Endocrinology and Metabolism
    Excerpt

    Recent clinical studies have questioned whether there is a qualitative change in the circulating isoforms of LH and FSH after stimulation by GnRH. The present study investigated the median charge of serum gonadotropin isoforms before and after an exogenous challenge of 100 micrograms GnRH in 10 girls and 10 boys undergoing pubertal development. All of the children had basal serum levels of LH and FSH and responses 30 and 60 min after GnRH treatment that were considered normal for puberty. LH and FSH in serum and eluates after electrophoresis in 0.10% agarose suspension were measured with sandwich fluoroimmunoassays. The increases in serum gonadotropin concentrations were generally similar for both sexes, but girls had significantly (P < 0.05) higher LH levels at 30 and 60 min than boys and a larger (P < 0.05) relative increase in serum FSH levels after GnRH treatment. In terms of the median charge of serum isoforms, the girls had significantly less negative (i.e. more basic) isoforms of LH (P < 0.05) and FSH (P < 0.001) than the boys. There was a change to more basic isoforms of both LH and FSH in all children 30 min after GnRH administration. For LH, the charge had returned to pretreatment values by 90 min after GnRH, but for FSH, the charge was still significantly (P < 0.05) more basic at this time (n = 4/sex). When the LH isoforms were more acidic before GnRH treatment, the change in median charge was larger than when the isoforms were more basic beforehand. A similar relationship was not found for FSH. Conversely, there was for FSH, but not for LH, a significant (P < 0.001) relationship between the relative increase in serum concentrations and the change in charge of the isoforms 60 min after GnRH treatment. These findings show that the circulating forms of LH and FSH become more basic after an exogenous challenge of GnRH in children undergoing pubertal development and suggest that the differences in the responses of LH and FSH isoforms may be due to differing degrees of selective secretion and/or survival.

    Title Tumor Necrosis Factor-alpha Downregulates Protein S Secretion in Human Microvascular and Umbilical Vein Endothelial Cells but Not in the Hepg-2 Hepatoma Cell Line.
    Date August 1994
    Journal Blood
    Excerpt

    Protein S deficiency, which is associated with thrombosis, can either be inherited or acquired. Recently, we reported that a decrease in free protein S was observed in 19 of 25 persons with HIV/AIDS. The proinflammatory cytokine, tumor necrosis factor-alpha (TNF-alpha), has been reported to be elevated in human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) patients and has been shown to induce a procoagulant state on the surface of endothelial cells. We report here that recombinant TNF-alpha (rTNF-alpha) downregulated protein S synthesis in the SV-40T transfected human microvascular endothelial cell line (HMEC-1) model system by approximately 70% and in primary human umbilical vein and dermal microvascular endothelial cell cultures by approximately 50%. Using the HMEC-1 model, Northern blot analysis showed a decrease in protein S RNA at 24 hours that was corroborated by Western blot analysis and enzyme-linked immunosorbent assay (ELISA) quantification. Evidence supporting the specificity of the TNF-alpha effect included the following: (1) TNF-alpha down-regulation of protein S was completely blocked by TNF neutralizing antibody; (2) the effect was transient, and protein S was restored to near normal levels after TNF was removed from cell cultures; (3) an antibody directed to the TNF RI (55-kD receptor) was shown to mimic the action of TNF-alpha on HMEC-1 cells; and (4) other proinflammatory cytokines, interleukin (IL)-1, IL-6, and TGF-beta, had no effect on protein S secretion. However, TNF-alpha showed no regulatory control over protein S synthesis in the human hepatocellular carcinoma cell line HepG-2. We suggest that TNF-alpha downregulation of protein S may be a mechanism for localized procoagulant activity and thrombosis recently reported in some AIDS patients with associated protein S deficiency.

    Title Ovaries of Ewes Homozygous for the X-linked Inverdale Gene (fecxi) Are Devoid of Secondary and Tertiary Follicles but Contain Many Abnormal Structures.
    Date February 1994
    Journal Biology of Reproduction
    Excerpt

    Ewes homozygous (II) for the Inverdale prolificacy gene (FecXI) located on the X chromosome are infertile with "streak" ovaries. The aims of this study were to examine the ovarian morphology and plasma and tissue (or extracellular fluid) hormone concentrations in II ewes in comparison to control (++) animals. All II animals (n = 11) were found to contain no normal ovarian follicles beyond the primary stage of development despite their being similar in number of germ cells to the ++ ewes. The II animals had high plasma gonadotropin levels and undetectable estradiol and progesterone, and in addition 6 of 9 ewes had undetectable plasma inhibin concentrations. II ewes contained large numbers of oocyte-free follicles ("nodules") that were often found as clusters in the innermost regions of the cortex. In addition, 6 of 11 ewes contained abnormal luteal- or granulosa cell-like structures and other unusual formations. In three II animals the abnormal structures were visible on the ovarian surface, and the plasma inhibin concentrations in these ewes were significantly higher than in the ++ ewes (p < 0.05). Collectively these findings suggest that two copies of the FecXI mutation impair the resumption of growth of primordial follicles and may in some instances lead to the development of abnormal structures having a morphology consistent with that observed for ovarian tumors.

    Title Biopotency in Vitro and Metabolic Clearance Rates of Five Pituitary Preparations of Follicle Stimulating Hormone.
    Date January 1994
    Journal Reproduction, Fertility, and Development
    Excerpt

    Five pituitary preparations of follicle stimulating hormone (FSH), namely NIDDK-oFSH-17, Bioscan oFSH, Ovagen, Folltropin-V and F.S.H.-P., were examined for biological activity in terms of their potency in an in vitro bioassay, receptor assay and heterologous radioimmunoassay and in terms of their metabolic clearance rates. In the three assays, Bioscan oFSH was the most potent (P < 0.05) (3- to 5-fold the potency of NIDDK-oFSH-17), with Ovagen being 25-50% the potency of the NIDDK standard (P < 0.05). Folltropin-V and F.S.H.-P. had the lowest potencies in all three assays. For each preparation, the ratio of activities between the assays was not consistent, suggesting that the preparations behaved differently in each assay. In 9 of 10 cases, potency estimates in the heterologous radioimmunoassay were greater than those in the in vitro bioassay or receptor assay. Polyacrylamide gel electrophoresis of the preparations showed banding consistent with the molecular weight of FSH, but also indicated that the preparations were contaminated with other proteins to varying extents. The half-lives of these preparations when injected into the bloodstream of mature female mice were 28.0, 8.6, 13.4, 11.6 and 17.4 min for NIDDK-oFSH-17, Bioscan oFSH, Ovagen, Folltropin-V and F.S.H.-P. respectively. The slopes of the decay rates were significantly different from each other (P < 0.05) except between Ovagen and Folltropin-V. The results of these studies show that a number of widely available FSH preparations have differing biopotencies. Moreover, the biopotency of a preparation in vitro is not related to its metabolic clearance rate, and not all FSH preparations behave identically in different assays. Measures of biopotency in vitro combined with those of metabolic clearance rate may provide useful information on the properties of FSH preparations used for research purposes and for superovulation of farmed livestock.

    Title Differential Patterns of Dynamic Cardiovascular Regulation As a Function of Task.
    Date December 1993
    Journal Biological Psychology
    Excerpt

    In cardiovascular reactivity studies, interpretations of the processes supporting the blood pressure response may become problematic when systolic blood pressure, diastolic blood pressure, and heart rate all increase in response to a behavioral challenge. Therefore, in addition to evaluating these cardiovascular responses, this study examined cardiac output, total peripheral resistance and systolic time intervals derived from impedance cardiogram, electrocardiogram and phonocardiogram recordings during a speech stressor, a mirror tracing task, and a foot cold pressor test. All of the behavioral stressors elicited increases in blood pressure and heart rate, with the largest changes occurring during the overt speech. Based on the examination of the response patterns of the underlying hemodynamic variables it would appear that, in both men and women, the blood pressure increase during the speech preparation period was supported by increased cardiac output; the speech itself resulted in a mixed pattern of increased cardiac output and total peripheral resistance; whereas, the mirror tracing and cold pressor tasks produced increased total peripheral resistance. Although men and women produced similar response patterns to the behavioral challenges, sex differences in the estimates of myocardial contractility were observed during rest. These results provide evidence that different behavioral stressors can produce a distinct yet integrated pattern of responses, whose differences may be revealed, when impedance cardiography is used, to derive sufficient response measures for assessing dynamic cardiovascular processes.

    Title Effects of Ovariectomy and Genotype on Bioactive Fsh in Plasma and Pituitary of Booroola Ewes.
    Date November 1993
    Journal Journal of Reproduction and Fertility
    Excerpt

    Blood samples were collected for 13 days before and 20 days after ovariectomy from carrier (BB) and non-carrier (+ +) ewes of the Booroola FecB gene (n = 12 per genotype), at known stages of the oestrous cycle, after which the pituitary glands from these ewes were recovered. Pituitary glands were also collected from cyclic ewes (about day 12; n = 5 per genotype) to compare the effects of ovariectomy on pituitary gonadotrophins. Plasma samples and pituitary extracts were assayed for bioactive (B) FSH, immunoreactive (I) FSH and I-LH. Overall, BB ewes had significantly (P < 0.05) higher plasma I-FSH concentrations than did + + ewes before ovariectomy; the mean value was higher on 16 of the 17 days of the oestrous cycle (P < 0.01). For B-FSH, there were no overall genotypic differences, although the mean for the BB ewes was significantly higher on 13 of the 17 days of the oestrous cycle (P < 0.05), and significantly (P < 0.05) higher between days 13 and 16. No genotypic differences were noted for the plasma bioactive:immunoreactive (B:I) ratio for FSH before ovariectomy. After ovariectomy, there were significant (P < 0.001) increases in plasma for B-FSH, I-FSH and I-LH and a significant (P < 0.05) decrease in the B:I ratio for FSH, irrespective of genotype.(ABSTRACT TRUNCATED AT 250 WORDS)

    Title Effects of the Booroola Gene (fecb) on Body Weight, Ovarian Development and Hormone Concentrations During Fetal Life.
    Date September 1993
    Journal Journal of Reproduction and Fertility
    Excerpt

    The aim of this study was to determine whether the FecB gene influenced some aspects of fetal development in sheep. Carrier (BB/B+) and non-carrier (++) female fetuses were recovered at specific times of gestation, namely, days 40, 55, 75, 90, 95 and 135. The results showed that the FecB gene influenced litter size, body weight and ovarian development during fetal life. The mean litter sizes were larger (P < 0.05) and body weights were lighter (P < 0.05) at most gestational ages in BB/B+ than in ++ fetuses. Morphometric studies of the ovary showed that the development of the BB/B+ ovaries was retarded: the ++ genotype had more oogonia at day 40 (P < 0.01), more germ cells entering meiosis at day 55, more primordial follicles developing at days 75, 90 and 95 (P < 0.05), a greater loss of germ cells by atresia at day 90 (P < 0.01) and more growing follicles (P < 0.01) and more antral follicles (P < 0.05) at day 135. Differences between the BB/B+ and ++ genotypes in the plasma concentrations of immunoreactive (i) inhibin, i-FSH, bioactive (b)-FSH or (i)-LH were not apparent at any age except for i-LH at day 75 (BB/B+ > ++; P < 0.05). Likewise no differences were noted in the contents of ovarian or adrenal oestradiol or i-inhibin except for i-inhibin in the adrenal at day 75 (++ > BB/B+, P < 0.01). No differences between the genotypes were noted in the i-inhibin contents of the mesonephros at day 40. In mid- to late but not early gestation (i.e. days 40 and 55) significant correlations (i.e. P < 0.05) were noted between litter size and body weight at days 75, 90 and 135, and between litter size and ovary weight, ovary volume, adrenal weight and pituitary weight at day 135. To eliminate the effect of litter size, equal numbers of BB/B+ and ++ embryos were transferred to respective recipient ewes, and fetuses were recovered at the equivalent of days 40 and 90 of gestation. The results showed that the genotypic difference in fetal body weight at day 40 (++ > BB, P < 0.001) and in number of oogonia at day 90 (++ > BB/B+, P < 0.05) were independent of litter size.(ABSTRACT TRUNCATED AT 250 WORDS)

    Title Gonadotrophin-releasing Hormone and the Control of Ovulation Rate by the Fecb Gene in Booroola Ewes.
    Date September 1993
    Journal Journal of Reproduction and Fertility
    Excerpt

    Booroola sheep carry a FecB gene that confers high fecundity. The aims of the present studies were to determine in homozygous carriers (BB) and non-carriers (++) of the Booroola FecB gene whether there are FecB differences in the secretory characteristics of GnRH in hypophyseal-portal blood of ovariectomized ewes (Expt 1) and whether differences in ovulation rate would occur following the administration of PMSG and pulsatile GnRH to ovary-intact Booroola ewes with hypothalamic-pituitary disconnection (HPD) (Expt 2). In Expt 1, no FecB gene-specific differences were noted for GnRH with respect to pulse frequency, pulse amplitude or overall secretion rate. Irrespective of genotype there were 1.9 +/- 0.2 GnRH pulses h-1 (n = 20 ewes; 10 BB and 10 ++ animals) and 1.9 +/- 0.1 pulses of immunoreactive (i) LH h-1. In Expt 2, ovulation was induced in the HPD ovary-intact animals on three occasions (e.g. 56, 393 and 423 days after HPD surgery) using a PMSG (200 or 400 iu)-GnRH pulse (250 ng i.v. for 96 h)-GnRH bolus (10 micrograms i.v.) regimen after pretreating the animals with GnRH pulses (250 ng i.v. for 14 days). On all occasions the ovulation rates were significantly higher (P < 0.05) in BB ewes (n = 6 or 7) than in ++ animals (n = 7). No differences between the genotypes were noted with respect to the mean concentrations of progesterone in plasma notwithstanding the differences in ovulation rates.(ABSTRACT TRUNCATED AT 250 WORDS)

    Title Protein S, an Antithrombotic Factor, is Synthesized and Released by Neural Tumor Cells.
    Date July 1993
    Journal Journal of Neurochemistry
    Excerpt

    Protein S, an anticoagulant factor in the protein C antithrombotic pathway, was found to be synthesized and released by six tumor cell lines of neural origin by western blotting and ELISA. The rate of synthesis ranged from three- to 11-fold higher than that of a microvascular endothelial cell line and 36-144% that of a hepatoma cell line. The secreted protein S displayed specific anticoagulant activity similar to that of purified plasma protein S, implying that it was fully gamma-carboxylated. Ten primary brain tumor tissues also expressed protein S antigen, as shown by western blot analysis. Expression of anticoagulantly active protein S by neural cells raises important questions concerning possible physiologic roles for this multidomain protein beyond its function in control of thrombosis.

    Title And the Beat Goes On.
    Date March 1993
    Journal Cranio : the Journal of Craniomandibular Practice
    Title Pcr Regimen for Enhanced Specificity and Yield of Targeted Genomic Dna Sequences: Ras and P53.
    Date February 1993
    Journal Pcr Methods and Applications
    Excerpt

    By weighting the PCR reaction in favor of specificity for the target sequence in the beginning cycles and for continued efficient amplification of the sequence into later cycles, we were able to show an improvement in the specificity and quantity of amplified ras and p53 sequences. Increased purity and yield of specific products favorably enhanced post-PCR evaluation and interpretation of results using direct sequencing and single-stranded conformation polymorphism (SSCP) analysis when point mutations were present in DNA from tumor cell lines and tissues.

    Title Bioactive and Immunoreactive Fsh and Immunoreactive Inhibin Concentrations in the Ovine Fetus.
    Date October 1992
    Journal The Journal of Endocrinology
    Excerpt

    The bioactive (B) and immunoreactive (I) pituitary contents/concentrations of FSH, together with the plasma concentrations of B-FSH, I-FSH and I-inhibin were determined in ovine fetuses at days 55, 75, 90 and 135 of gestation (day 145 = term). The pituitary contents and concentrations of B-FSH and I-FSH increased in both sexes with gestational age. The female fetuses had significantly (P < 0.01) higher pituitary contents/concentrations of B-FSH and I-FSH than the male fetuses at days 75 and 135. The pituitary B/I ratios of FSH were not significantly different with age or sex. The plasma concentrations of B-FSH remained relatively constant from days 75 to 135, with no significant differences between sexes or with age. In contrast, the plasma concentrations of I-FSH reached a peak at day 90 and then declined towards term in both sexes. At all gestational ages except day 55, the female fetuses had significantly (P < 0.05) higher plasma concentrations of I-FSH than the males. In both sexes, the plasma B/I ratios of FSH were lowest at day 90 and had increased again by day 135, with the male fetuses having significantly (P < 0.05) higher B/I ratios compared with the female group at days 75 and 135 but not at day 90. At all gestational ages, the plasma concentrations of I-inhibin declined throughout gestation in the female fetuses, whereas in the males they reached a nadir at day 75 and then increased towards term. The concentrations of I-inhibin were significantly (P < 0.01) higher in the male fetuses compared with the females.(ABSTRACT TRUNCATED AT 250 WORDS)

    Title Flow Separation in the Carotid Bulb: Prognostic Significance.
    Date October 1992
    Journal Neurological Research
    Excerpt

    It has been previously shown that boundary layer or flow separation occurring in the carotid bulb and detected by duplex scanning denotes minimal or no carotid atherosclerotic disease as demonstrated by angiography and reliably predicts aetiology other than carotid artery disease in symptomatic patients. To evaluate outcome at long-term follow-up we prospectively studied 94 patients (48 males, 46 females) who demonstrated bilateral flow separation. Mean age was 61.2 years (27 to 86 years). Mean follow-up was 57 months (5 to 113 months). There was one death during follow-up at 69 months. It was stroke related. Using age and sex specific death rates for the general population 14.3 deaths would be expected for the same average period. By life table analysis, survival was 98.7% at five years compared to a general population expected 5 year survival of 85.9%. There were no strokes at 5 years of follow-up. (Age and sex specific stroke-free survival for Rochester, MN 1970-1974 is 98% at 5 years). TIA-free survival was 99% at one year (n = 87) and 96% at five years (n = 46). It is concluded that the presence of boundary layer separation in the carotid bulb not only indicates absent or minimal atherosclerotic disease, but is predictive of a favourable long-term outcome with respect to mortality and neurological events.

    Title Evaluation of Monoclonal Antibodies Having Specificity for Human Igg Subclasses: Results of the 2nd Iuis/who Collaborative Study.
    Date March 1992
    Journal Immunology Letters
    Excerpt

    Following the 1st IUIS/WHO Collaborative Study of monoclonal anti-IgG subclass antibodies, a panel of WHO Specificity Reference Reagents (SRR) was established [Jefferis, R., et al. (1985) Immunol. Lett., 10, 223]. At the time, the hope was expressed that further reagents particularly for IgG2, and other allotypic specificities would become available which could be applied in a wide range of assay protocols. The 2nd study reports the evaluation of nineteen anti-subclass and seven anti-allotype monoclonal antibodies. The anti-IgG1 antibody HP6187 was equivalent in performance to the SRR. Others, that were not of the mouse IgG1 isotype, may be useful for particular applications. The anti-IgG2 antibody HP6200 could be a valuable addition to the WHO SRR; it is specific for an epitope in the Fab region but does not have the light chain bias of HP6014. Antibodies of putative allotype specificity exhibited the claimed specificity when used within protocols similar to those employed by the originating laboratory. It appears to be inherent in the nature of the epitopes (allotopes) recognized that it will take several years before reagents applicable to a wide range of techniques will become available.

    Title Effects of Modifying Gonadotrophin-releasing Hormone Input Before and After the Oestrogen-induced Lh Surge in Ovariectomized Ewes with Hypothalamo-pituitary Disconnection.
    Date January 1991
    Journal The Journal of Endocrinology
    Excerpt

    The patterns of gonadotrophin-releasing hormone (GnRH) input to the pituitary gland that affect the expression of a positive-feedback event by oestrogen on LH secretion were investigated in ovariectomized ewes with hypothalamo-pituitary disconnection (HPD). In experiment 1, ovariectomized HPD ewes were given hourly i.v. pulses of 250 ng GnRH and an i.m. injection of 50 micrograms oestradiol benzoate (OB). The ewes were given a bolus pulse of 2.25 micrograms GnRH 16 h after injection of OB, followed by half-hourly pulses of 250 ng GnRH for 14 h (treatment A). The LH surge response was significantly (P less than 0.05) greater in these ewes compared with that in ewes given a continuous infusion of GnRH (250 ng/h) after the OB injection, followed by a continuous infusion of 500 ng GnRH/h after the bolus pulse of GnRH (treatment B). When no GnRH was administered after the OB injection, except for the bolus pulse of GnRH (treatment C), the surge response was significantly (P less than 0.05) reduced compared with that in treatment A, and was reduced compared with treatment B. These data suggest that GnRH pulses are important in the generation of the OB-induced LH surge, but that a baseline secretory component can prime the pituitary to some extent. In experiment 2, a doubling of the continuous infusion dose of GnRH used in treatment B to 500 ng/h before the bolus pulse of GnRH and to 1 micrograms/h afterwards (treatment D) gave a similar response compared with treatment A, suggesting that if the baseline input of GnRH is of sufficient magnitude, it can overcome the lack of pulsatile input. In experiment 3, halving the GnRH pulse amplitude used in treatment A from 250 to 125 ng (treatment E) did not reduce the LH surge response, implying that when the GnRH input is in a pulsatile mode, the amplitude of GnRH pulses is less important than the pulsatile nature per se. In experiment 4, removal of GnRH input after the bolus pulse of GnRH (treatment F) significantly (P less than 0.05) reduced the surge response compared with when pulses were maintained (treatment A), indicating that GnRH input is still required once the LH surge has been initiated. Collectively, these experiments show that several forms of GnRH delivery, both pulsatile and baseline, can result in the full expression of a positive-feedback response in ovariectomized ewes treated with oestrogen.

    Title A Triangulation Method for the Quantitative Measurement of Arterial Blood Velocity Magnitude and Direction in Humans.
    Date December 1990
    Journal Ultrasound in Medicine & Biology
    Excerpt

    A triangulation method has been applied to a duplex ultrasound scanner to quantify blood flow velocities in two dimensions. A position locating system (PLS) connected to the scanhead locates the sample volume (SV) in 3-D space to a precision of 1 mm. The PLS is used to obtain flow velocity data from two independent lines of sight in the human femoral artery. Data are gathered from anatomic sites of interest along one line of sight. Later the computer directs the SV to interrogate the same points in space from a second line of sight. Water tank studies using both constant velocity and pulsatile string targets were used to validate the method. Velocity magnitudes could be calculated to within 5% error for Doppler angles below 75 degrees for various string depths and speeds; the error in Doppler angle calculation was usually less than 3 degrees. Results from the superficial femoral artery show flow velocity vectors are nearly parallel to the vessel walls. Peak systolic velocity magnitudes range from 63-66 cm/s in three presumed normal individuals. Following the validation studies addressed in this paper, this triangulation approach is intended in future work to document the complex nonaxial character of blood flow that occurs normally at branch points and in regions of intraluminal disease.

    Title Effects of the Synthetic Glucocorticoid Dexamethasone on Reproductive Function in the Ewe.
    Date October 1990
    Journal The Journal of Endocrinology
    Excerpt

    Glucocorticoids have been found to inhibit reproductive function in most domestic species studied but, in the ewe, preliminary reports suggest that glucocorticoids may have little or no effect. This study investigated the effects of the synthetic glucocorticoid dexamethasone on oestrus and ovulation rate in ewes during the breeding season and gonadotrophin secretion in the breeding and non-breeding seasons. In cyclic ewes, dexamethasone treatment at rates of up to 2 mg/day did not affect the natural or pregnant mare serum gonadotrophin-stimulated ovulation rate, or the timing and incidence of behavioural oestrus (P greater than 0.05). Dexamethasone administration (2 mg/day) had no effect on LH secretion or the plasma LH response to a 1 micrograms injection of gonadotrophin-releasing hormone (GnRH) in ovariectomized ewes in the breeding and non-breeding seasons, and did not compromise the inhibition of plasma LH levels during chronic treatment with oestrogen. Similarly, dexamethasone had no effect on plasma FSH concentrations, but significantly (P less than 0.05) reduced the plasma FSH response to a 1 micrograms GnRH injection during chronic negative treatment with oestrogen in ovariectomized ewes. Collectively, these data show that in these experiments dexamethasone did not significantly modify reproductive function in the ewe, a finding that is in contrast to that found in other domestic species.

    Title Testing Ultrasonic Pulsed Doppler Instruments with a Physiologic String Phantom.
    Date October 1990
    Journal Journal of Ultrasound in Medicine : Official Journal of the American Institute of Ultrasound in Medicine
    Excerpt

    The spatial, temporal, and frequency resolution of conventional ultrasonic Doppler instruments and the time/space distortions in two-dimensional color Doppler imaging systems can be measured using a pulsatile moving string target. The diameter of the string is small compared with the Doppler sample volume, the velocity (speed and direction), acceleration and timing of the string motions are precisely known with reference to the R wave timing mark, and the spatial location of the string is known. A loop of surgical thread or monofilament fishline running between pulleys is driven by a motor that provides constant string speeds from 0.05 to 150 cm/s and variable string speeds programmed to mimic arterial velocity waveforms from the carotid, aortic, and femoral arteries. Constant string speeds are used to evaluate the Doppler sensitivity, frequency processing, and sample volume size; pulsatile movement of the string provides a physiologic model to evaluate the temporal performance of conventional Doppler systems and the temporal and spatial performance of two-dimensional color Doppler imaging scanners.

    Title Recent Advances in Carotid Artery Evaluation.
    Date May 1990
    Journal Clinics in Diagnostic Ultrasound
    Excerpt

    This chapter was intended to provide an overview of ultrasound duplex scanning at the human carotid bifurcation. Although results of clinical studies are useful in patient management, much remains to be learned in the area of understanding the disease process and determining what features, obtainable by ultrasound, will predict clinical outcome. Listed below are the important points covered by this chapter. 1. Ultrasound duplex scanning can detect and quantify carotid artery disease with respect to the gold standard of contrast arteriography. 2. Contrast arteriography is not the ideal gold standard for assessment of cartotid artery disease. A new standard incorporating attributes of not only anatomy but also physiology is needed. 3. Using currently available ultrasound systems it is not possible to obtain the Doppler angle (i.e., the angle formed by the transducer axis and the blood flow-velocity vector). This angle is often estimated by the angle formed by the transducer axis and the axis of the blood vessel. This assumes that the flow-velocity vectors are axial, which turns out to be a poor assumption at bifurcations, curves, and sites of intraluminal disease. Specifying the Doppler angle involves assumptions that should always be kept in mind. 4. All Doppler data should be collected at the same angle formed by the sound beam relative to the vessel axis. Whereas results of Doppler studies can be reported in units of either frequency or velocity, assumptions made about the Doppler angle and acquiring data at arbitrary angles can produce large errors when quantifying flow velocities. 5. Blood flow disturbances cannot automatically be equated with the presence of disease. Some disturbances are localized secondary blood flow patterns produced by changes in vascular geometry coupled with pulsatile flow. 6. The lack of blood flow disturbances cannot automatically be equated with a normal or disease-free artery. Accumulation of plaque within the sinus region at the minimal and moderate disease stages appears to eliminate or attenuate local flow disturbances characteristic of a normal bifurcation. 7. An understanding of the blood flow patterns at clinical sites of interest is important to the correct interpretation of data. Both the imaging and pulsed Doppler modalities are essential to this understanding: the imaging capability to place the pulsed Doppler sample volume and appreciate changes in vascular anatomy, and the pulsed Doppler to obtain the character of blood flow at specific sites within the blood vessels.

    Title An Ultrasonic Measurement of Superficial Femoral Artery Wall Thickness.
    Date March 1990
    Journal Ultrasound in Medicine & Biology
    Excerpt

    An ultrasonic measurement of the superficial femoral artery wall thickness was performed on 16 volunteers. The measurement included all echogenic tissue between the lumen of the superficial femoral artery and the lumen of the superficial femoral vein. The average arterial wall plus vein wall thickness in volunteers with peripheral arterial disease was 2.13 +/- 0.87 mm, significantly greater than the 1.27 +/- 0.50 mm found in those without detectable peripheral arterial disease.

    Title Recommended Guide to the Evaluation of Permanent Impairment of the Temporomandibular Joint.
    Date March 1990
    Journal Cranio : the Journal of Craniomandibular Practice
    Excerpt

    This paper provides a method to evaluate the extent of permanent injury to the temporomandibular joint. The authors, working as the Committee on Permanent Impairment for the American Academy of Head, Facial, Neck Pain and TMJ Orthopedics, have used the same values as the American Medical Association has used in their Guides to the Evaluation of Permanent Impairment for other disk-protected and functional joints. These Guides may then serve as a tool to help physicians and dentists who treat temporomandibular joint injuries to rate the degree of permanent injury. These Guides may be combined with other areas of impairment that are already described and valued in the Guides to the Evaluation of Permanent Impairment published by the AMA. These Guides provide a range and each injury should be evaluated on an individual basis. We urge you to use these Guides conservatively and in harmony with good clinical judgement.

    Title The Oestrogen-induced Surge of Lh Requires a 'signal' Pattern of Gonadotrophin-releasing Hormone Input to the Pituitary Gland in the Ewe.
    Date October 1989
    Journal The Journal of Endocrinology
    Excerpt

    Two experiments were conducted with ovariectomized and hypothalamo-pituitary disconnected (HPD) ewes to ascertain the pattern of inputs, to the pituitary gland, of gonadotrophin-releasing hormone (GnRH) necessary for the full expression of an oestrogen-induced LH surge. The standard GnRH replacement to these sheep was to give pulses of 250 ng (i.v.) every 2h; at the onset of experimentation, pulses were given hourly. In experiment 1, groups of sheep (n = 7) were given an i.m. injection of 50 micrograms oestradiol benzoate, and after 10 h the GnRH pulse frequency or pulse amplitude was doubled. Monitoring of plasma LH concentrations showed that a doubling of pulse frequency produced a marked increase in baseline values, whereas a doubling of amplitude had little effect on the LH response. In a second experiment, ovariectomized HPD sheep that had received hourly pulses of GnRH for 16 h after an i.m. injection of oil or 50 micrograms oestradiol benzoate were given either a 'bolus' (2.25 micrograms GnRH) or a 'volley' (500 ng GnRH pulses 10 min apart for 30 min, plus a 500 ng pulse 15 min later). Both groups then received GnRH pulses (250 ng) every 30 min for the next 13 h. Oestrogen enhanced the LH responses to the GnRH treatments, and the amount of LH released was similar in ovariectomized HPD ewes given oestrogen plus bolus or volley GnRH treatments and ovariectomized hypothalamo-pituitary intact ewes given oestrogen. These results suggest that the oestrogen-induced LH surge is initiated by a 'signal' pattern of GnRH secretion from the hypothalamus.

    Title Organochlorine Chemicals in Human Breast Milk in Hong Kong.
    Date October 1989
    Journal Archives of Environmental Contamination and Toxicology
    Excerpt

    The results of a small-scale survey of organochlorine contaminants in human breast milk are presented and compared to previous data from 1976 for Hong Kong, and to reported data from elsewhere. Concentrations of p,p DDT, dieldrin and hexachlorobenzene (HCB) in recent breast milk samples were slightly but significantly lower than those in samples taken a decade previously in Hong Kong. However, levels of gamma-HCH (hexachlorocyclohexane) were higher in the more recent samples. The concentrations of DDT, DDE and beta-HCH in Hong Kong breast milk remain among the highest reported in the literature. Probable routes of uptake of such contaminants include the ingestion of seafood, as these same compounds have previously been found at high concentrations in mussels from Hong Kong waters. Data for organochlorine levels in breast milk from other parts of Asia are not available, but trends in global pesticide usage suggest that significant contamination elsewhere in the continent is likely.

    Title Specificity and Association Constants of 33 Monoclonal Antibodies to Human Iga Epitopes.
    Date September 1989
    Journal Immunology Letters
    Excerpt

    We used an immunofluorescent sequential-saturation-of-antibody assay and an interactive computer program for Scatchard analysis to determine association constants (Ka) of 33 murine monoclonal antibodies (Mabs) specific for human IgA epitopes. Ka ranged from 0.37 to 690 x 10(7) liters per mole (an approximate 1900-fold difference). Specificity was validated with a panel of 18 highly purified IgA1 and IgA2 myeloma proteins and secretory IgA using an immunofluorometric assay. Western blots of bacterial IgA protease digests were used to locate the epitopes of IgA specific Mabs in either the Fab, Fc, or hinge region. Mabs specific for unique epitopes on secretory IgA or free secretory component (FSC) were produced and evaluated.

    Title Should Results of Ultrasound Doppler Studies Be Reported in Units of Frequency or Velocity?
    Date July 1989
    Journal Ultrasound in Medicine & Biology
    Excerpt

    There is a current tendency to report the results of ultrasound Doppler studies in units of velocity instead of Doppler frequency. This is probably motivated by the intuitive feeling that blood flow studies should naturally be reported in cm/s and the notion that "velocity" is a normalizing factor for Doppler ultrasound studies. In order to determine velocity, the Doppler angle theta or angle formed by the ultrasound beam and flow velocity vector, must be known. It is not possible, using currently available systems, to obtain an accurate estimate of this angle. The physics related to the Doppler equation are reviewed in this paper along with examples to illustrate the origin and magnitude of errors that could arise when reporting in units of velocity. Guidelines are provided for thinking about and reporting results of Doppler studies in units of velocity. An understanding of the Doppler equation and its use in clinical studies are promoted in this paper to enhance the diagnostic usefulness of Doppler ultrasound studies and to reduce serious errors which could lead to faulty information dictating patient management.

    Title Diagnostic Significance of Flow Separation in the Carotid Bulb.
    Date April 1989
    Journal Stroke; a Journal of Cerebral Circulation
    Excerpt

    Pulsatile blood flow within the normal carotid sinus involves at least two distinct components. That near the flow divider is laminar and antegrade, whereas a boundary layer separation zone in the posterolateral aspect exhibits transient blood flow reversal. It is now possible to document these flow velocity components using pulsed Doppler ultrasound methods. When atherosclerosis develops, it preferentially involves the posterolateral bulb region, obliterating the normal configuration of the sinus with consequent loss of the flow separation zone. It was therefore hypothesized that if flow separation could be detected, it should be predictive of a normal angiogram. To assess this, we evaluated 20 symptomatic patients and two with only bruits found by duplex scanning to have flow separation in either one or both carotid bulbs and who also underwent cerebral angiography. Initial diagnoses were stroke in seven, reversible ischemic neurologic deficit in one, transient ischemic attack in 12, and bruit in two. Flow separation was bilateral in 13 patients (59%). There were 15 patients with symptoms in the territory of a carotid bulb exhibiting flow separation. By angiography, of the 35 bulbs with boundary layer separation, 27 (77%) were normal, with the remainder showing lesions that reduced the diameter of the vessel by 20% or less. Final diagnoses of the 15 patients with symptoms ipsilateral to a carotid sinus exhibiting flow separation were fibromuscular disease in two, lacunar stroke in three, dissection in two, subclavian steal in one, cardiogenic embolus in three, migraine in one, hyperventilation syndrome in one, kink of the mid-internal carotid artery in one, and no diagnosis in one.(ABSTRACT TRUNCATED AT 250 WORDS)

    Title Estimation of Association Constants of 42 Monoclonal Antibodies to Human Igg Epitopes Using a Fluorescent Sequential-saturation Assay.
    Date May 1988
    Journal Immunology Letters
    Excerpt

    We measured the association constant (Ka) for 42 murine monoclonal antibodies (MAbs) to human IgG epitopes. Included are antibodies, previously evaluated in an IUIS/WHO collaborative study, to various epitopes on the four subclasses of human IgG - IgG Fc, IgG Fab, kappa, and lambda - and to selected IgG allotopes. We used a sequential-saturation immunofluorescent assay and interactive computer program to determine the Ka by Scatchard analysis. Kas ranged from unmeasurably low by this method (approximately 10(6) L/M) to 3.8 X 10(9) L/M. Some class specific MAbs had large Ka differences for different subclasses and some subclass specific MAbs had large Ka differences for molecules of the same subclass but of different light-chain types.

    Title Variability in Measurement of Specific Parameters for Carotid Duplex Examination.
    Date January 1988
    Journal Ultrasound in Medicine & Biology
    Excerpt

    The variability of four carotid artery frequency parameters used for classifying disease with duplex scanning was prospectively studied. Forty-eight patients (94 patent carotid arteries) were each examined by two technologists. Measured parameters were the peak systolic frequency (PSF) and the first zero slope from the common carotid artery, and the PSF and end diastolic frequency (EDF) from the internal carotid artery. Measurements from all the examinations were made twice by each technologist. Interobserver, intraobserver, and interpatient variability in measurement of the first zero slope was so great that we have abandoned its use. Measurement of variability for PSF and EDF was much less (correlation coefficients 0.68 to 0.92). These parameters were measured with sufficient precision to warrant their continued use for important decision steps in classifying carotid artery disease. Interpatient differences in PSF sufficient to cause disagreement regarding the hemodynamic significance of carotid disease occurred in only three instances. In each of these cases the differences were due to examination technique (failure to identify a very distal internal carotid artery stenosis, difficulty distinguishing between a kink and a stenosis, and failure to recognize an improper Doppler angle). We conclude that the variability of PSF and EDF is within clinically acceptable levels and is mainly due to examination technique rather than measurement of waveform parameters or changes in patient hemodynamics.

    Title Noninvasive Assessment of Normal Carotid Bifurcation Hemodynamics with Color-flow Ultrasound Imaging.
    Date November 1987
    Journal Ultrasound in Medicine & Biology
    Excerpt

    The combination of a B-mode imaging system and a single range-gate pulsed Doppler flow velocity detector (duplex scanner) has become the standard noninvasive method for assessing the extracranial carotid artery. However, a significant limitation of this approach is the small area of vessel lumen that can be evaluated at any one time. This report describes a new duplex instrument that displays blood flow as colors superimposed on a real-time B-mode image. Returning echoes from a linear array of transducers are continuously processed for amplitude and phase. Changes in phase are produced by tissue motion and are used to calculate Doppler shift frequency. This results in a color assignment: red and blue indicate direction of flow with respect to the ultrasound beam, and lighter shades represent higher velocities. The carotid bifurcations of 10 normal subjects were studied. Changes in flow velocities across the arterial lumen were clearly visualized as varying shades of red or blue during the cardiac cycle. A region of flow separation was observed in all proximal internal carotids as a blue area located along the outer wall of the bulb. Thus, it is possible to detect the localized flow patterns that characterize normal carotid arteries. Other advantages of color-flow imaging include the ability to rapidly identify the carotid bifurcation branches and any associated anatomic variations.

    Title Determining Total Cost-effectiveness of Drug Therapy.
    Date April 1987
    Journal American Journal of Hospital Pharmacy
    Title A Comprehensive Hospital Pharmacy Program of Intrapreneurial Ventures.
    Date March 1987
    Journal Topics in Hospital Pharmacy Management / Aspen Systems Corporation
    Title Isotype-specific Enzyme Immunoassay for Influenza Antibody with Monoclonal Antibodies to Human Immunoglobulins.
    Date January 1987
    Journal Journal of Clinical Microbiology
    Excerpt

    Mouse monoclonal antibodies specific for human immunoglobulin isotypes were investigated for use in an isotype-specific enzyme immunoassay for detection of antibody to influenza type A hemagglutinin (H1 and H3). The monoclonal antibody reagents were compared with isotype-specific, hyperimmune rabbit antisera from the National Institutes of Health. Endpoint titers for immunoglobulin G (IgG) obtained with the two reagents were within fourfold of each other 84% of the time (79 of 84) and within eightfold of each other 95% of the time (89 of 94). Regression analysis of the data gave a multiple correlation coefficient (r2) of 0.77 and a Spearman rank value of 0.83 (P less than 0.001). For IgA reagents, endpoint titers agreed within fourfold 77% of the time (88 of 114) and within eightfold 92% of the time (105 of 114). The r2 was 0.73, and Spearman rank was 0.83 (P less than 0.001). IgM antibody was detected in only 17 of 114 sera by either monoclonal or polyclonal reagents. Of these sera, 14 (82%) gave titers with the two reagents that were within fourfold of each other. A similar number of fourfold titer rises were detected with each reagent in paired sera showing hemagglutination inhibition titer rises. Monoclonal antibody reagents detected 27 IgA, 29 IgG, and 6 IgM rises, while polyclonal antisera detected 26 IgA, 31 IgG, and 7 IgM rises. These results show that monoclonal antibodies specific for human immunoglobulin isotypes are suitable as reagents for diagnostic assays. The advantages of monoclonal antibodies are their high degree of specificity and the ability to be standardized and produced in unlimited quantities. Moreover, the availability of immunoglobulin subclass- and allotype-specific monoclonal antibodies will enable a more detailed analysis of the antibody response to influenza as well as other infectious agents.

    Title Collective Bargaining Should Be a Last-ditch Effort.
    Date August 1986
    Journal American Journal of Hospital Pharmacy
    Title Diastolic Flow As a Predictor of Arterial Stenosis.
    Date April 1986
    Journal Journal of Vascular Surgery : Official Publication, the Society for Vascular Surgery [and] International Society for Cardiovascular Surgery, North American Chapter
    Excerpt

    With a pulsed Doppler imaging system, it is now possible to interrogate sites from the aorta to the popliteal trifurcation. To determine which velocity parameters could be correlated with the degree of disease as determined by angiography, 34 arterial stenoses identified by scanning were also evaluated by contrast arteriography and classified in 10% increments. The angiographic readings were blinded with respect to the scan results. Four hand-measured parameters from velocity waveforms obtained at the site of stenosis were correlated with the angiogram--peak systolic velocity, systolic rise time, diastolic reverse velocity, and diastolic reverse flow time. When diastolic reverse flow was absent, diastolic forward flow was recorded. To describe diastolic flow along a continuum, diastolic reverse velocity was ascribed a positive value and diastolic forward velocity was ascribed a negative value. A systolic velocity gradient (peak velocity/rise time) was also calculated. The relationship between the angiographic categories and the measured parameters was evaluated with the Jonkheere-Terpstra trend test. A trend was determined with diastolic flow (diastolic reverse flow or diastolic flow velocity) that was significant (p less than 0.01). The linear regression was calculated (y = 40.8 + [-5.6X]), and correlation coefficient was obtained (r = 0.76) that was statistically significant (p less than 0.01). The method enables mapping and calculation of arterial stenoses by noninvasive means. This can be expected to obviate the need for diagnostic angiograms in certain and select cases in which angioplasty can be expected to be beneficial. It also affords a convenient quantitative means of following lesions over time.

    Title Human Resource Management in Hospital Pharmacy.
    Date December 1985
    Journal Topics in Hospital Pharmacy Management / Aspen Systems Corporation
    Title Evaluation of Monoclonal Antibodies Having Specificity for Human Igg Sub-classes: Results of an Iuis/who Collaborative Study.
    Date November 1985
    Journal Immunology Letters
    Excerpt

    Seventy-four monoclonal antibodies (McAb) of putative specificity for human IgG (11), the IgG sub-classes (59) or Gm allotypes (4) have been evaluated for reactivity and specificity in eight laboratories employing different assay techniques or protocols. For the IgG, IgG3, IgG4, G1m(f) and G3m(u) specificities McAb have been produced that can be satisfactorily applied in most methodologies employed and have potential as reference reagents. The IgG1 and particularly IgG2 specificities proved problematical with all McAb evaluated demonstrating apparent assay restriction and whilst performing well in some assays proved to be poor or inactive reagents in others. However, the study identifies McAb individually suited to application within most commonly employed methodologies. Epitope display is the probable variability rather than capricious behaviour by the McAb. IgG1 and IgG2 were the least immunogenic of the sub-class proteins and there is evidence that epitope display is influenced by the physical and chemical procedures used to immobilize or fix antigen - a common requirement in the assay systems studied.

    Title Sources of Variability in Carotid Duplex Examination: a Prospective Study.
    Date November 1985
    Journal Ultrasound in Medicine & Biology
    Excerpt

    Interobserver and intraobserver variability of ultrasonic, duplex carotid artery examinations was studied in a prospective, randomized, and blinded clinical trial. Forty-eight patients were examined by two technologists, yielding 96 carotid artery examinations. The kappa statistic was calculated to determine the degree of agreement corrected for chance. The kappa value between examinations by different technologists was 0.476. Variability occurred at both steps in the examination procedure: (1) obtaining the velocity waveforms (kappa = 0.536); and (2) using these waveforms to classify the extent of carotid disease (kappa = 0.609 for interobserver variability in reading waveforms). Minimal to moderate disease categories accounted for most of the variability. There was little disagreement in categorizing lesions as greater than or less than 50% diameter reduction. Intraobserver variability in rereading spectral waveforms was minimal (kappa = 0.842 and 0.894). Recognition of disturbed flow patterns in normal carotid bulbs may reduce variability.

    Title Noninvasive Mapping of Lower Limb Arterial Lesions.
    Date November 1985
    Journal Ultrasound in Medicine & Biology
    Excerpt

    Thirty patients with peripheral arterial disease were evaluated using an ultrasonic duplex scanner. A total of 338 arterial segments from the level of the iliac to the popliteal artery were studied and compared with the results of arteriography read independently by two radiologists who were unaware of the results with the scanner. The results demonstrate that this method is not only suitable for clinical use but is as good as arteriography in defining both the location and extent of the arterial involvement.

    Title Immunoassay for Igg Rheumatoid Factor with a Murine Monoclonal Anti-fd Antibody.
    Date August 1985
    Journal The Journal of Laboratory and Clinical Medicine
    Excerpt

    Conventional radioimmunoassays for IgG rheumatoid factors (RF) detect the binding of IgG RF (or their F(ab')2 fragments) to solid-phase human IgG Fc fragments or rabbit IgG. Binding is detected with a radiolabeled antibody that is IgG-specific but is nonreactive with human Fc (i.e., an alpha-Fd reagent) or with rabbit IgG. Anti-Fd reagents are quite laborious to produce. We developed a murine monoclonal alpha-Fd antibody, confirmed its specificity, and compared it with conventional rabbit alpha-Fd in the IgG RF radioimmunoassay. We also adapted the assay to an enzyme-linked assay with both human Fc and rabbit IgG as antigen. We compared the four assay methods with each other and examined their relationship to certain clinical and laboratory features of rheumatoid arthritis.

    Title Hemodynamics of the Normal Human Carotid Bifurcation: in Vitro and in Vivo Studies.
    Date August 1985
    Journal Ultrasound in Medicine & Biology
    Excerpt

    The spatial and temporal characteristics of blood flow in the normal adult human carotid bifurcation are investigated by two different methods: in vitro pulsatile flow model experiments using laser Doppler anemometry and in vivo studies employing pulsed Doppler velocity measurements obtained with an ultrasound duplex scanner. Glass and Plexiglas models based upon arteriographic measurements were evaluated with laser Doppler anemometer methods for pulsatile flow. A similarity approach permits the model study to be geometrically and hydrodynamically accurate with respect to the human carotid bifurcation. These parallel but separate approaches were originally performed by the principal authors without knowledge of each others' work. Normal flow patterns in the proximal internal carotid artery are demonstrated to include: unidirectional, helical, transient reversal, and low velocity regions of flows. The characterization of these complex temporal and spatially variant flow fields required the high sample volume resolution afforded by the model study. Pulsed Doppler ultrasound and a novel method of positioning the sample volume permitted a qualitative description of the complex flow velocity fields in the normal human bifurcation. Results of the two methods are compared and a striking similarity between the two methods is observed for the primary and secondary flow features. The problem of associating blood flow velocity disturbances with the presence of intralumenal disease is addressed in the discussion. It is suggested that the flow disturbances associated with the normal carotid bifurcation are different from those associated with intraluminal disease and further, that the secondary flow structures can be usefully employed to establish normalcy.

    Title Carotid Endarterectomy. Relationship of Outcome to Early Restenosis.
    Date June 1985
    Journal Journal of Vascular Surgery : Official Publication, the Society for Vascular Surgery [and] International Society for Cardiovascular Surgery, North American Chapter
    Excerpt

    The results following carotid endarterectomy were prospectively evaluated in 134 patients (145 sides) by repeat ultrasonic duplex scanning and clinical evaluation extending for a period of 4 years. There were 107 men and 27 women in the study group. The perioperative stroke rate was 1.3% and the mortality rate, 0.7%. There were 9 late deaths, of which two were stroke related (1.4%). Focal symptoms occurred in 12 patients on the ipsilateral side, six of which were strokes (one lacunar). The remaining symptoms developed in the presence of moderate degrees of carotid stenosis (less than 50%). There were seven patients who had transient ischemic attacks (TIAs) referable to the operated side, but only two of these were associated with a recurrent high-grade stenosis. During follow-up 32 (22%) patients had recurrent high-grade stenosis. Restenosis regressed in seven, giving a persistent rate of 17.1%. The incidence of restenosis was significantly higher in women (p less than 0.01). By life-table analysis, restenosis occurred early, the majority within 24 months. There was no consistent association between the development of symptoms and the occurrence of restenosis. Therefore, it is concluded that there is no justification for reoperation based on the degree of narrowing observed to prevent subsequent TIAs and strokes.

    Title Igg Subclass Antibodies to Herpes Simplex Virus.
    Date June 1985
    Journal The Journal of Infectious Diseases
    Excerpt

    Several mouse monoclonal antibodies specific for human IgG1, IgG2, IgG3, and IgG4 were evaluated by enzyme-linked immunosorbent assay to detect human IgG subclass antibodies to herpes simplex virus (HSV) antigens. The variable results with different monoclonal antibodies point to the need for well-characterized reagents in the study of antibody responses to infectious agents. The 204 sera tested were obtained from 157 patients with various forms of clinically manifest HSV infections and from several controls. IgG1 antibodies were demonstrated in almost all HSV-infected subjects and were the first antibodies to appear in primary genital infections. IgG2, IgG3, and IgG4 antibodies were detected in acute-phase sera, most often in patients with recurrent genital herpes but in none of those with primary infections. IgG4 antibodies occurred significantly more frequently in sera from men than in those from women with recurrent genital infections.

    Title Cryptosporidiosis in Hospital Personnel. Evidence for Person-to-person Transmission.
    Date May 1985
    Journal Annals of Internal Medicine
    Excerpt

    An intern responsible for the care of a patient with chronic cryptosporidiosis developed acute diarrhea and serologic evidence of cryptosporidium infection. Sera from 26 hospital personnel exposed to the patient and 18 personnel with no exposure were examined with an indirect immunofluorescent antibody procedure for the presence of antibodies to Cryptosporidium. Eight (31%) exposed personnel--5 nurses, 2 house officers, and 1 student--had positive antibody titers (1:10 or more). The frequency of positivity in the nurse-housestaff-student group (8 of 18, 45%) was significantly greater (p less than 0.05) than that in the attending physicians and respiratory therapists (0 of 8). The former group had significantly more exposure to the patient's feces than did the latter group (p less than 0.01). Three of eighteen control personnel (17%) had positive cryptosporidium antibody titers. These findings suggest that Cryptosporidium may be transmitted from person to person in the hospital environment and that serologic evidence of infection is common among hospital personnel.

    Title Computer Based Pattern Recognition of Carotid Artery Doppler Signals for Disease Classification: Prospective Validation.
    Date April 1985
    Journal Ultrasound in Medicine & Biology
    Excerpt

    A computer based pattern recognition method has been developed to classify the percent diameter reduction in nonoccluded internal carotid arteries. Using a combined B-mode/pulsed Doppler unit, the system utilizes spectral waveforms obtained from the low common and proximal internal carotid artery locations. The ECG-R wave is used as a time reference to synchronize the averaging of Doppler spectra from 20 heart cycles. An averaged waveform is generated and represents the spectral data from which features are extracted for analysis. A stepwise selection algorithm identifies a feature subset for partitioning the entire range of disease into two states, less than and greater than a decision point. Three such partitions are made, leading to the following categories: Normal, 1-20, 21-50 and 51-99% dia. reduction. A classifier was trained, tested prospectively against unknown data and the results compared to angiography. Of the 170 vessels tested, 141 (82%) were classified in the same category by angiography and the computer system. Agreement for each category was 93% (27/29) for the normals, 81.5% (44/54) for the 1-20% lesions, 78% (29/37) for the 21-50% lesions and 82% (41/50) for the 51-99% lesions. The computer method and angiography differed by more than one category in only one of the 170 tests. The level of agreement corrected for chance (Kappa +/- SE(K] was 0.769 +/- 0.039. Future efforts will be directed toward dividing classification of disease further (especially in the 51-99% category), developing a dedicated microprocessor for on-line analysis of the signals and using the system for prospective epidemiological studies of various populations.

    Title Six-month Hospital-pharmacy-based Technician Training Program.
    Date February 1985
    Journal American Journal of Hospital Pharmacy
    Excerpt

    A six-month training program for hospital pharmacy technicians is described. The development of this program was stimulated by the need to use departmental resources more efficiently and the desire of local hospital pharmacy directors to have trained technicians. Financial support was sought and received from the local Private Industry Council. The department selects 10 students and two alternates for each class on the basis of test performance and an interview. The program consists of 1014 scheduled hours of instruction. The first week is spent orienting the students to the program and general hospital pharmacy concepts. The next six weeks are spent in a laboratory where the students acquire their basic skills. The rest of the program, approximately 4.5 months, is spent in on-the-job training, additional didactic lectures, and field trips. The students are evaluated routinely throughout the program. At the end of the program, each participant is granted a certificate. Attrition has been low; three classes have completed the program, and 28 technicians have graduated. Twenty-five of these technicians are now employed full-time. Positive feedback has been received from the technicians' employers. This program meets the need of area hospital pharmacies to recruit trained technicians.

    Title Evaluation of Thirty-one Mouse Monoclonal Antibodies to Human Igg Epitopes.
    Date January 1985
    Journal Hybridoma
    Excerpt

    Stable clones of 31 mouse hybridomas that produce monoclonal antibodies (MAbs) against human IgG antigenic determinants were obtained. The number of hybridomas of different specificity described are: 2 anti-IgG1 Fc, 1 anti-IgG2 Fc, 1 anti-IgG2 Fd, 2 anti-IgG3 Fc, 2 anti-IgG3 hinge, 3 anti-IgG4 Fc, 3 anti-IgG4 Fd, 2 anti-nG4m(b), 4 anti-IgGFc, 2 anti-IgGFd, 1 anti-kappa, 1 anti-lambda, 1 anti-non IgG1, 2 anti-non IgG2, 2 anti-non-IgG3, 2 anti-non-IgG4. Evidence is presented validating their specificity. Some MAbs demonstrated to be avid, potent, and specific for well defined IgG-subclass epitopes may be partially or completely inactive in other assay systems, presumably because of different presentations of antigen epitopes. In general, this problem requires careful writing of protocols describing the use of MAbs.

    Title Diversification Strategies for Hospital Pharmacies.
    Date November 1984
    Journal American Journal of Hospital Pharmacy
    Excerpt

    Several ways used by the pharmacy department of a large university hospital to generate revenue through diversification are described. The department offers its facilities and staff as a resource in training medical service representatives for several pharmaceutical manufacturers, which is projected to provide $85,000 in net income for fiscal year (FY) 1983-84. The pharmacy department also conducts a six-month program for training pharmacy technicians, which yields a small net profit. The pharmacy department actively participates in educational programs such as college courses and clerkships earning extra income. An apothecary-style outpatient pharmacy was set up under a for-profit corporation. Services have been expanded to include the preparation of i.v. solutions that support home care. A durable medical equipment (DME) business is planned. The ambulatory and home-care programs are expected to generate approximately $165,000 in net profit next year. Contract pharmaceutical services are provided to another hospital. The net income generated through diversification in this pharmacy department will exceed $250,000 in FY 1983-84.

    Title Natural History of Carotid Artery Disease on the Side Contralateral to Endarterectomy.
    Date November 1984
    Journal Journal of Vascular Surgery : Official Publication, the Society for Vascular Surgery [and] International Society for Cardiovascular Surgery, North American Chapter
    Excerpt

    The natural history of the nonoperated carotid artery opposite an endarterectomy was examined in 134 patients by means of ultrasonic duplex scanning over a period extending to 48 months. None of the nine deaths that occurred during follow-up was stroke related. A total of 22 arteries showed progression of disease over this period. By life-table analysis the mean annual rate of progression for all categories of disease was 12.6% and 7.4% for progression to a diameter reduction greater than 50%. Disease progression was more rapid in patients under 65 years of age. Symptoms occurred in 13 patients for an overall incidence of 10% and a mean annual rate estimated at 5%. All symptoms indicated transient ischemic attacks; there were no strokes. There was a strong relationship between the development of symptoms and stenoses greater than 80% either at the initial examination or secondary to progression. No correlation was found between the presence of bruits or their change over time and the progression or appearance of symptoms. Conservative management of nonoperated vessels opposite an endarterectomy appears appropriate until symptoms develop or a lesion greater than 80% is detected.

    Title A Collage of Multi-disciplinary Techniques in the Advanced Case of Periodontal Prosthesis.
    Date October 1984
    Journal Periodontal Case Reports : a Publication of the Northeastern Society of Periodontists
    Title The Natural History of Carotid Arterial Disease in Asymptomatic Patients with Cervical Bruits.
    Date August 1984
    Journal Stroke; a Journal of Cerebral Circulation
    Excerpt

    A prospective study was initiated in January 1980 to follow with Duplex scanning a consecutive series of 167 asymptomatic patients with cervical bruits. Patients were seen at six month intervals for the first year and yearly thereafter. Based on previously validated criteria, disease at the carotid bifurcation was classified into 6 categories: Normal, 1-15% diameter reduction, 16-49%, 50-79%, 80-99%, and occlusion. Patients were evaluated to assess: the occurrence of new neurological symptoms, the stability of the lesions at the carotid bifurcation, and the possible role of risk indicators on disease changes. During follow-up, ten patients became symptomatic (6 with TIA's and 4 with stroke). The development of symptoms was accompanied by disease progression in 8 patients. By life table analysis, the annual rate occurrence of symptoms was 4%. The mean annual rate of disease progression to a greater than 50% stenosis was 8%. When progression in all categories was considered, 60% of the sides showed some disease aggravation. The presence of or progression to a greater than 80% stenosis was highly correlated (p = 0.00001) with either the development of a total occlusion of the internal carotid artery or new symptoms. The major risk factors associated with disease progression were cigarette smoking, diabetes mellitus, and age. Those patients under 65 years of age were most likely to show progression. Despite high rates of disease progression, this study further supports the contention that it is prudent to follow a conservative course in the management of asymptomatic patients presenting with a cervical bruit.(ABSTRACT TRUNCATED AT 250 WORDS)

    Title Phase I Study of Intravenous Gamma Globulin in Multiple Myeloma.
    Date May 1984
    Journal The American Journal of Medicine
    Excerpt

    Seventeen patients with multiple myeloma were given intravenous immunoglobulin at doses ranging from 150 mg/kg to 500 mg/kg in a phase I study. The intravenous immunoglobulin was well tolerated with only three transient episodes of mild clinical toxicity during 27 infusions. In no instance was hepatic or renal toxicity seen. Marked biologic variability over the one month study period in total IgG levels in patients with non-IgG myeloma and IgG subclasses in many of the patients was observed, making intravenous immunoglobulin half-life determinations based on IgG or IgG subclass levels problematical. The decay of functional antibody to hepatitis B surface antigen was determined. Analysis of the hepatitis antibody data suggested that intravenous immunoglobulin half-life was in the range of seven to 20 days for the entire study group and was not related to the isotype of the myeloma paraprotein or to the baseline levels of IgG. No infections were observed in the study group during the study period, but the potential for infection prophylaxis by intravenous immunoglobulin in myeloma patients must be evaluated in a randomized, prospective, controlled phase III study.

    Title Application of a Solid-phase Immunofluorometric Assay to the Selection of Monoclonal Antibody Specific for the Adenovirus Group-reactive Hexon Antigen.
    Date April 1984
    Journal Archives of Virology
    Excerpt

    An immunofluorometric assay (IFMA) has been evaluated as a screening assay to detect monoclonal antibodies to the group-specific antigen of the adenovirus hexon component. The antibodies were produced as mouse ascitic fluids from hybridoma cells generated from Balb/C mice immunized with purified adenovirus type 2 hexon component and crude adenovirus type 3 culture supernatants. The purified IgG fractions from all monoclonal ascitic fluids tested were identified as the IgG1 K mouse isotype. Antibody titers ranged from 102,400 to 204,800 by the IFMA, from 200 to 12,800 by indirect FA, and were generally nonreactive in counterelectrophoresis, complement fixation, hemagglutination-inhibition, serum neutralization, and immune electron microscopy titrations. The IFMA is a reliable method for quantitating low levels of specific antibody in large numbers of test samples, and is therefore ideal as a screening assay for monoclonal antibody in tissue culture fluids and in mouse ascitic fluids.

    Title The Impact of Duplex Scanning on the Evaluation of Patients with Asymptomatic Bruits in the Region of the Carotid Arteries.
    Date December 1983
    Journal The Netherlands Journal of Surgery
    Excerpt

    Between January 1980 and January 1981, 95 patients with 'asymptomatic' bruits in the region of the carotid arteries were evaluated at the University of Washington with an ultrasonic Duplex scanner combined with spectral analysis. Based upon the interpretation of a hardcopy of the spectral analysis, the artery of interest was classified into one of five categories: A normal; B 5-15% diameter reduction; C 16-49% diameter reduction; D 50-99% diameter reduction, and E total occlusion. Forty-eight patients had unilateral and 47 patients had bilateral bruits; 142 bruits were evaluated. The distribution of disease in the internal carotid artery was: A: 2% (3/142); B: 30% (42/142); C: 37% (53/142); D: 29% (41/142), and E: 2% (3/142). If the presence of a high grade stenosis of the internal carotid artery (50-99% diameter reduction) is an indication for invasive intervention, then 71% of these patients would not require contrast arteriography. The role of a direct ultrasonic study, such as Duplex scanning, in patients with asymptomatic bruits is discussed.

    Title Hospital-based Training for Pharmaceutical Manufacturers' Representatives.
    Date December 1983
    Journal American Journal of Hospital Pharmacy
    Excerpt

    A hospital-based training program for pharmaceutical manufacturers' representatives is described. The pharmacy department of a large teaching institution established a training program for new sales representatives of a major pharmaceutical company. Goals were outlined by the sales training manager and the pharmacy department. The sales training personnel, department of pharmacy, and the cooperating departments of medicine, surgery, and nursing worked together to formulate objectives for the sessions, and teaching responsibilities were delegated to members of all these departments. The program length varied from one to five days. A formal contract was developed specifying content, program dates, and reimbursement. The institution is reimbursed for the use of the facility, materials, and administrative overhead. The program's success has led to the development of similar programs with several other companies. The extra income has enabled the pharmacy to create a new division within the department. Evaluations from more than 500 sales representatives who have participated in the programs have been consistently positive. The pharmacy department in a teaching institution has the resources to provide a training program for sales representatives that can be an additional source of income.

    Title The Surgical Management of the Restorative Alveolar Interface (ii).
    Date November 1983
    Journal The International Journal of Periodontics & Restorative Dentistry
    Title Flow Velocity Patterns in the Carotid Bifurcations of Young, Presumed Normal Subjects.
    Date September 1983
    Journal Ultrasound in Medicine & Biology
    Excerpt

    Spectral analysis of pulsed Doppler velocity waveforms has been found useful as a diagnostic technique in the assessment of carotid artery disease. While spectral broadening of the velocity waveform obtained at center stream sites is usually associated with arterial disease, the present study describes spectral patterns resulting from disturbed blood flow in the proximal branches of the carotid bifurcation in young, presumed normal human subjects. In those studied, spectral patterns in the bifurcation region exhibit characteristics similar to those occurring in zones of flow separation in model studies under conditions of steady flow. It is important to distinguish the spectral patterns due to arterial disease from those occurring in the normal bifurcation. This paper describes the types of flow disturbances noted in presumed normal arteries and points out the need to understand the flow velocity patterns that may be found at specific anatomical sites across the carotid bifurcation.

    Title Evaluating Carotid Artery Disease. The Concordance Between Pulsed Doppler/spectrum Analysis and Angiography.
    Date September 1983
    Journal Ultrasound in Medicine & Biology
    Excerpt

    The results of ultrasonic pulsed Doppler duplex scanning with spectral analysis and computer pattern recognition are compared with the results of contrast arteriography in patients screened for extracranial carotid artery disease. The intraangiographer variability (one radiologist reading the same films twice) and the interangiographer variability (two radiologists reading the same film independently) were also studied. To calculate degrees of agreement corrected for chance, the Kappa statistic was computed for all the evaluation methods employed. At the present time, the concordance between spectral analysis and cerebral contrast angiography reaches a Kappa value of 0.682 +/- 0.064. This level of agreement compares favorably with the interangiographer agreement level (K = 0.568 +/- 0.058) and the intraangiographer agreement (K = 0.711 +/- 0.054). The computer pattern recognition program predicted the degree of stenosis by angiography with an agreement of K = 0.721 +/- 0.059. This concordance compares favorably to that observed when the radiologists are compared with themselves and is greater than that reached by two different radiologists. The continuous improvement in precision and accuracy of duplex scanning offers the promise of its usefulness in clinical and epidemiological studies.

    Title Post-endarterectomy Carotid Ultrasonic Duplex Scanning Concordance with Contrast Angiography.
    Date September 1983
    Journal Ultrasound in Medicine & Biology
    Excerpt

    The results of ultrasonic duplex scanning combined with spectral analysis are compared with the results of contrast angiography in patients after endarterectomy in which recurrence of carotid arterial disease was suspected. Thirty-six patients underwent a duplex scan study within 3 months of their post-operative angiogram, performed at their physician's discretion (44 studies). The overall accuracy of the method was 80%. Our ability to predict a greater than 50% diameter reduction along with total occlusion was 94%. The measure of agreement corrected for chance between arteriography and duplex scanning as expressed by the Kappa statistic was 0.675 +/- SE (K) 0.096. This level of agreement compared favorably to that of inter- and intra-observer variability in reading cerebral angiograms. The accuracy reported justifies the clinical use of ultrasonic duplex scanning in the detection of recurrent stenosis after carotid endarterectomy.

    Title Trace Elements in the Pacific Oyster in Hong Kong.
    Date January 1983
    Journal Archives of Environmental Contamination and Toxicology
    Title Computer Based Classification of Carotid Arterial Disease: a Prospective Assessment.
    Date December 1982
    Journal Stroke; a Journal of Cerebral Circulation
    Excerpt

    A minicomputer based pattern recognition method has been used to prospectively classify the category of disease involvement of 105 carotid arteries. The system utilized spectral patterns obtained from a combined B-mode/pulsed Doppler unit. All decisions are based upon comparison of an unknown, averaged waveform with a series of vessels with known severity of disease. The variability in the computer decision as compared to arteriography is discussed.

    Title Evaluation of a Solid-phase Immunoassay with Fluorescein Isothiocyanate-conjugated Heterogeneous or Monoclonal Antibodies for Identification of Virus Isolates, with Influenza Virus As a Model.
    Date September 1982
    Journal Journal of Clinical Microbiology
    Excerpt

    A solid-phase immunofluorescence assay was evaluated for the identification of viruses isolated in tissue culture, with influenza virus as a model. Purified immunoglobulin G (IgG) from hyperimmune rabbit sera specific for contemporary strains of influenza A or B was covalently attached to microscopic plastic beads to capture virus. Fluorescein isothiocyanate (FITC)-conjugated antibodies of different specificities were then reacted with bound antigen, and the resulting complexes were quantified in a suitable filter fluorometer. The assay, with appropriately absorbed FITC-conjugated second antibody, reliably identified virus present in harvests from cell cultures infected with clinical specimens. For influenza A (H1N1) virus, sensitivity of detecting antigen was about 8- to 32-fold less when an FITC-conjugated monoclonal Igg antibody pool specific for epitopes in three different antigenic sites on influenza hemagglutinin was used as the second antibody as compared to when IgG from hyperimmune sera specific for virus or its components was used as the second antibody. The immunofluorometric assay provides a method for quantitative detection of viral antigen in tissue culture fluids and objective identification of virus type and subtype with FITC-conjugated reagents.

    Title Immunoglobulin M and G Antibody Response to Type- and Subtype-specific Antigens After Primary and Secondary Exposures of Mice to Influenza A Viruses.
    Date August 1982
    Journal Infection and Immunity
    Excerpt

    A mouse model of influenza infection was studied to help define parameters that may affect serodiagnosis of human infections by immunoassays. Antibodies to both type- and subtype-specific influenza A antigens were measured by a solid-phase immunofluorometric assay. Dilute mouse sera were added to purified influenza virus that had been covalently bound to polyaminostyrene microbeads, and the bound antibody was detected by fluorescein isothiocyanate-labeled isotype-specific antisera. Results were consistent in that upon exposure of mice by either infection alone or by vaccination after infection, both immunoglobulin M (IgM) and IgG antibodies reactive with newly encountered subtype specific viral antigens were measured. IgG antibody was usually detectable by the solid-phase immunofluorometric assay several days before it could be detected by a hemagglutination inhibition test. Increased levels of antibody of the IgG1, IgGa, IgG2b, and IgG3 subclasses were also measured during influenza infection. Surprisingly, response to type-specific viral antigens was of the IgG class in primary as well as in secondary exposure. The results suggest that for serodiagnosis of influenza infections by detection of specific IgM antibody, the assay should use subtype-specific antigens.

    Title Empirical Findings Relating Sample Volume Size to Diagnostic Accuracy in Pulsed Doppler Cerebrovascular Studies.
    Date July 1982
    Journal Journal of Clinical Ultrasound : Jcu
    Excerpt

    Ultrasonic duplex scanning has a sensitivity of 97% in detecting disease of the carotid bulb. However, the specificity is much less, being 37%. The error in distinguishing normal arteries from those with minimal disease appears to be in part related to the size of the sample volume of the pulsed Doppler. This paper addresses the importance of the features of scan head design in relation to distinguishing normal and diseased arteries.

    Title Word Processing in Hospitals: Improve Information Handling While Holding Down Costs.
    Date June 1982
    Journal Hospital Financial Management
    Title Computer Based Pattern Recognition of Carotid Arterial Disease Using Pulsed Doppler Ultrasound.
    Date June 1982
    Journal Ultrasound in Medicine & Biology
    Excerpt

    A minicomputer based system has been developed for studying carotid artery blood flow data obtained for a combined B-mode, pulsed Doppler ultrasound scanner. The goals of this work are to devise and improve techniques for estimating the extent of atherosclerosis at the carotid artery bifurcation. Features are automatically extracted from spectrum analyzed Doppler blood flow data. Five statistical pattern recognition algorithms are compared, with cross validation being used to improve the estimate of classification accuracy. A data collection protocol has been devised in which four sites are studied along each carotid arterial system. Classification of unknowns is done using a hierarchy of three decisions.

    Title Spectral Analysis of Doppler Velocity Patterns in Normals and Patients with Carotid Artery Stenosis.
    Date May 1982
    Journal Clinical Physiology (oxford, England)
    Excerpt

    A computerized pattern recognition program was utilized to assess the predictive ability of various parameters obtained from the spectra of ultrasonic pulsed Doppler signals from the carotid arteries. The most accurate features selected by linear regression analysis were the natural log (ln) of the ratio of the mean velocity in the internal carotid artery compared to that in the common carotid artery, and the ln of the maximum velocity, the ln of the maximum frequency, and the square of the fractional broadening term, all of which were measured at peak systole in the internal carotid artery. Using the combination of the velocity ratio and the fractional broadening term, the average difference in the estimated percentage stenosis, as compared to that obtained by arteriography, was 12.8%.

    Title Evaluating Doppler Devices Using a Moving String Test Target.
    Date March 1982
    Journal Journal of Clinical Ultrasound : Jcu
    Excerpt

    Two moving string test targets have been developed and used to characterize ultrasound Doppler instrumentation. The characteristics investigated were sample volume size and location in space, amplitude sensitivity to Doppler shift, and resolution of Doppler direction. The tests were performed on a combined echo/Doppler instrument and on an annular array system. The procedures can be carried out routinely in the clinical laboratory to ensure that the instrument is working properly or as an aid for correct interpretation of acquired data.

    Title Evaluation of Carotid Bifurcation Disease: the Role of Common Carotid Artery Velocity Patterns.
    Date March 1982
    Journal Archives of Surgery (chicago, Ill. : 1960)
    Excerpt

    We evaluated duplex ultrasonographic velocity patterns from the common carotid artery in 156 patients with arteriographically verified internal carotid artery disease. Flow to zero during diastole in the common carotid was found in three groups of patients: (1) those with flow to zero in both common carotids not related to disease of the carotid bulb; (2) those with a tight (90% to 99%) stenosis in the internal carotid; and (3) those with total occlusion of the internal carotid. Although such flow to zero is commonly observed with total occlusion of the internal carotid artery, it may also be seen with high-grade stenosis. Thus, when found, it is essential to ascertain if flow is indeed present in the internal carotid artery; understanding changes in velocity patterns of the common carotid has improved our overall accuracy in detecting extracranial carotid artery disease.

    Title Ultrasonic Duplex Scanning for Disease of the Carotid Artery.
    Date January 1982
    Journal Circulation
    Excerpt

    The duplex ultrasonic scanner combines real-time B-mode imaging with a single-gate, variable-range pulsed Doppler. The detection and categorization of the severity of carotid artery atherosclerosis is achieved by performing spectral analysis of the pulsed Doppler velocity signal obtained from vessels of interest. Using this technique, 750 patients with suspected extracranial carotid artery disease were evaluated between January 1978 and January 1980. One hundred thirty-five of these 750 patients (18%) underwent cerebral arteriography performed with biplanar views of the carotid bifurcation. The degree of stenosis was measured independently in these patients and was available for comparison with the results of duplex scanning and spectral analysis. Duplex scanning correctly detected the presence of disease in 252 of 259 carotid arteries studied (97%). The extent of involvement varied from plaques that produced less than 10% diameter reduction to those that resulted in a total occlusion. The technique was less accurate with lesions that produced less than 10% diameter reduction.

    Title Effect of Test System on the Ability of Monoclonal Antibodies to Detect Antigenic Drift in Influenza A(h1n1) Virus Haemagglutinins.
    Date December 1981
    Journal The Journal of General Virology
    Excerpt

    Results of analysing antigenic variation in the haemagglutinin (HA) molecule of naturally occurring influenza A (H1N1) viruses from 1977 to 1979 with monoclonal antibodies were found to be dependent in some instances on the test system used. In several instances A/USSR/90/77 HA-specific monoclonal antibodies had sharply reduced haemagglutination-inhibition (HI) titres with variant virus although they bound to the variant and A/USSR/90/77 HAs with similar efficiencies as judged by titration in a sensitive and accurate solid-phase immunofluorimetric assay. In another instance, the converse situation was observed: monoclonal antibodies having a reduced efficiency of binding to the HA of a variant virus nevertheless had comparable HI titres with the variant and with A/USSR/90/77. The chemical basis and epidemiological significance of these observations remain to be elucidated. Nevertheless, the finding that the reaction of monoclonal antibodies can, in some cases, be markedly dependent on the test system employed is of significance for the efficient design and correct interpretation of immunochemical studies which employ monoclonal antibodies to investigate the basis for variation in influenza strains.

    Title Repair of Radiation-induced Dna Damage in Nondividing Populations of Human Diploid Fibroblasts.
    Date October 1981
    Journal Biophysical Journal
    Excerpt

    The occurrence of DNA repair in UV- (254 nm) and X-irradiated normal human diploid fibroblasts maintained in a quiescent, nondividing state using low serum (0.5%) medium was ascertained. Techniques that detect different steps of the excision repair process were used so that the extent of completion of repair at single sites could be determined. These included measuring the disappearance of pyrimidine dimers by chromatography, detecting repair synthesis by density-gradient and autoradiographic methods and detecting the rejoining of repaired regions and repair of x-ray-induced single-strand DNA breaks using alkaline sucrose gradients. Results show that dimer excision occurs and the subsequent steps of repair synthesis and ligation are completed. About 50% of the dimers formed by exposure to 20 J/m2 is excised in the initial 24-h post-UV period. DNA repair (unscheduled DNA synthesis) can be detected through a 5-d post-UV period. The fraction of damaged sites eventually repaired is not known. X-ray-induced single-strand DNA breaks are repaired rapidly.

    Title Noninvasive Detection of Internal Carotid Artery Occlusion.
    Date September 1981
    Journal Angiology
    Excerpt

    Flow in the common carotid artery is normally quasisteady with flow never approaching zero during diastole. With total occlusion of the internal carotid artery, flow in the common carotid assumes the pattern observed in the external carotid, which supplies a relatively high-resistance vascular bed. In 34 instances of total internal carotid obstruction, flow went to zero in diastole in 33 cases and also demonstrated flow reversal in 22. In addition, there was a significant reduction in peak systolic velocity when the low-resistance internal carotid was obstructed. These observations, which are simple to determine using an ultrasonic duplex scanner, are of value in suspecting total occlusion of the internal carotid artery, thus obviating the need for arteriography in some cases.

    Title Color Digital Echo/doppler Image Presentation.
    Date July 1981
    Journal Ultrasound in Medicine & Biology
    Title Detection of Monoclonal Influenza Antibodies Synthesized in Culture by Hybridoma Cells with a Solid-phase Indirect Immunofluorometric Assay.
    Date July 1981
    Journal Journal of Virological Methods
    Excerpt

    A solid-phase indirect immunofluorometric assay for measuring reactions of mouse monoclonal antibodies with antigen has been developed, with influenza virus as a model. Purified IgG from hyperimmune rabbit sera is covalently linked to polyaminostyrene beads, to which influenza viruses are then bound immunologically to make solid-phase antigens. Alternatively, the virus is covalently coupled directly to the beads. Mouse antibodies, produced by hybridoma cells in culture, are reacted with constant amounts of solid-phase antigens, and then indirectly quantitated by adding FITC-labeled antimouse Ig and measuring the fluorescent intensity with a filter-fluorometer. The assay system permits rapid screening for low levels of antibodies synthesized by hybridoma cells in culture. It is about 25- to 150-fold more sensitive than hemagglutination inhibition tests in detecting monoclonal antibodies reactive with influenza virion HA protein.

    Title Detection of Peripheral Vascular Disease Using the Duplex Scanner Iii.
    Date November 1980
    Journal Ultrasound in Medicine & Biology
    Title Quantitative Characterization of Specificity and Potency of Conjugated Antibody with Solid-phase, Antigen Bead Standards.
    Date September 1980
    Journal Journal of Immunological Methods
    Excerpt

    An immunofluorescent assay was used to characterize precisely the potency and specificity of fluorescein-conjugated immunoglobulin class-specific and polyvalent anti-sera. Stable antigen standards consisted of highly purified immunoglobulin antigens covalently bound to polyaminostyrene beads. Linear regressions of weighted logit transformations of relative fluorescent intensities versus the logarithm of relative conjugate concentrations were determined. The potency of conjugates was compared using two different methods derived from the logit transformation. Inappropriate specificities were measured in some conjugates described as immunoglobulin 'class-specific'.

    Title Ultrasonic Demonstration of External and Internal Carotid Patency with Common Carotid Occlusion: a Preliminary Report.
    Date September 1980
    Journal Stroke; a Journal of Cerebral Circulation
    Excerpt

    Non-invasive ultrasonic imaging of the carotid bifurcation by duplex scanning and ultrasonic arteriography combined with pulsed Doppler spectrum analysis demonstrated patency of the external and internal carotid arteries distal to a common carotid occlusion in 3 patients. Common carotid occlusion is not invariably associated with thrombosis of the ipsilateral internal carotid artery. Identification of internal carotid patency by the use of ultrasonic techniques will permit surgical treatment in selected cases.

    Title Carotid Artery Velocity Patterns in Normal and Stenotic Vessels.
    Date April 1980
    Journal Stroke; a Journal of Cerebral Circulation
    Excerpt

    Duplex scanning provides real time B-mode images of the carotid bifurcation vessels along with a single gate pulsed Doppler flow velocity detector. By using the B-mode output of the duplex system to measure the Doppler angle and spectrum analysis to measure the frequency content of the Doppler signal, instantaneous flow velocity can be calculated. Mean velocity at peak systole was calculated retrospectively in 68 common (CCA) and internal (ICA) carotid arteries of 39 patients who had undergone prior angiography and prospectively in 30 arteries of 15 healthy young controls. The ratio of mean peak ICA velocity to mean peak CCA velocity at systole (VICA/VCCA) was below 0.8 in all 36 normal arteries and above 1.5 in all 21 high-grade stenoses of 60% or greater diameter reduction. Sixty-one percent of 41 vessels with less than 10 to 55% diameter reduction had a velocity ratio between 0.8 and 1.5. Only 10% of all ICA's with any stenotic lesion were incorrectly classified as normal. VICA/VCCA appears to be an accurate indicator of the degree of ICA stenosis.

    Title Detection of Carotid Occlusive Disease by Ultrasonic Imaging and Pulsed Doppler Spectrum Analysis.
    Date December 1979
    Journal Surgery
    Excerpt

    Ultrasonic imaging of the cervical carotid arteries by ultrasonic arteriography and duplex scanning combined with pulsed Doppler spectrum analysis were investigated in a series of patients undergoing arteriography. By using the ultrasonic image as a guide for precise placement of the pulsed Doppler sample volume, the characteristics of blood flow at points of interest in the carotid arteries could be determined. Audible analysis of the Doppler signal permitted correct diagnosis of 23 of 26 (88%) high-grade stenoses or occlusions with ultrasonic arteriography and 24 of 26 (92%) with duplex scanning. Spectrum analysis of Doppler signals obtained with the duplex scanner detected all of the 22 high-grade stenoses. Spectral abnormalities of a lesser degree also were detected in 18 of 23 vessels (78%) with atherosclerotic plaques which should not have reduced cerebral blood flow. These techniques permit the accurate detection of and the distinction between high-grade stenoses and occlusion, as well as the identification of many plaques which are not large enough to affect intracranial hemodynamics.

    Title Pulsed Doppler Assessment of Normal Human Femoral Artery Velocity Patterns.
    Date September 1979
    Journal The Journal of Surgical Research
    Title The Significance of Carotid Bruits in Children: Transmitted Murmur or Vascular Origin, Studies by Pulsed Doppler Ultrasound.
    Date August 1979
    Journal American Heart Journal
    Excerpt

    Forty-four youngsters with precordial murmurs and carotid bruits were evaluated clinically and independently, using pulsed Doppler ultrasound. The precordial murmur was evaluated with M-mode echocardiography combined with Doppler flow evaluation, and the carotid bruit was evaluated with peripheral vascular sector scan with Doppler flow evaluation. These ultrasonic techniques can identify abnormal blood flow at anatomic sites such as the aortic valve and in the carotid arteries. The patients had no symptoms and their condition, except for six, was mild enough that catheterization was not indicated. The clinical diagnosis of aortic stenosis was made in 30 children, and nine were thought to have no heart disease. On the basis of the ultrasonic examinations, 28 patients were diagnosed as having aortic stenosis and seven subjects had no intracardiac turbulence. However, there was disagreement in 14 instances; four of the six clinical "normals" were found to have aortic stenosis by pulsed Doppler echocardiography; six patients diagnosed as having mild aortic stenosis on a clinical basis were found to have no aortic abnormality. The results confirm that aortic stenosis usually presents as a murmur maximal in the aortic area, which is associated with a carotid bruit. Unfortunately, in at least one-fourth of the cases the murmur was not maximal at the aortic area, and a carotid bruit was found in several normal subjects. Since the consequences of over- or under-diagnosis of aortic stenosis are substantial, careful thought should be given to the differential diagnosis, and if possible, pulsed Doppler echocardiography should be utilized for a definitive statement of aortic valve-induced turbulence.

    Title Antiserums for Immunofluorescent Enumeration of Human T Lymphocytes Utilizing Fluoresceinated Staphylococcal Protein A.
    Date October 1978
    Journal The American Journal of Pathology
    Excerpt

    Five lots (100 ml or more) of heterologous antiserums specific for human T lymphocytes were prepared using human or Rhesus monkey thymocytes as immunogens. After appropriate adsorptions, these antiserums reacted by immunofluorescence with 68% of human peripheral blood mononuclear cells and 98% of human thymocytes, with E-rosette--positive cells but not with EAC-rosette--positive cells or five human B-lymphoblastoid-cell lines. Blocking experiments showed that Rhesus monkey thymocytes share thymic antigenic determinant(s) with humans. E-rosette receptors modulated independently from T-cell heteroantigens. Non-E--rosetting neoplastic T cells were identified in several patients with lymphoproliferative malignancies. Applying both the E-rosette assay and the anti-T-cell serum provides a better method of defining the biologic properties of normal and neoplastic T lymphocytes. Standardization of immunofluorescent conjugates for human T- or B-cell enumeration is simplified if large lots of well-characterized antiserums are available.

    Title The Mononuclear Cell in Human Blood Which Mediates Antibody-dependent Cellular Cytotoxicity to Virus-infected Target Cells. Ii. Identification As a K Cell.
    Date April 1977
    Journal Journal of Immunology (baltimore, Md. : 1950)
    Excerpt

    Studies were carried out to determine whether the mononuclear cell in human blood which mediates antibody-dependent cellular cytotoxicity (ADCC) to herpes simplex virus (HSV)-infected target cells has surface Fc receptors which participate in the reaction. The F (ab')2 fragment of human IgG antibody was inactive both in ADCC and in complement-mediated cytolysis, but retained the capacity to neutralize infectious virus, to agglutinate erythrocytes coated with viral antigens, and to bind to the surface of virus-infected cells. Treatment of sensitized virus-infected target cells with staphylococcus protein A, which has affinity for the Fc epitope of IgG, strongly reduced their susceptibility to lysis by ADCC in a dose-dependent relationship. These findings indicate that the Fc portion of IgG antibody to the virus is necessary for cytotoxicity. Treatment of blood mononuclear cells with either heat-aggregated gamma-globulin or HSV immune complexes inhibited effector cell activity. The presence of "third party" cellular immune complexes also strongly inhibited ADCC. Adsorption of mononuclear cells to plastic surfaces coated with soluble third party immune complexes resulted in a significant reduction in effector cell activity. These findings demonstrate that the ADCC effector cell possesses surface Fc receptors which are utilized in the ADCC reaction. The presence of Fc receptors on the surface of the effector cell indicates that it is a K cell rather than a null cell.

    Title Analysis of Mononuclear Cell Surfaces with Fluoresceinated Staphylococcal Protein A Complexed with Igg Antibody of Heat-aggregated Gamma-globulin.
    Date January 1977
    Journal Journal of Immunology (baltimore, Md. : 1950)
    Excerpt

    Fluorescein-conjugated staphylococcal protein A (SPA) was complexed with either: 1) heat-aggregated IgG, 2) B cell specific antibody, or 3) T cell specific antibody and then used for an immunofluorescent analysis of mononuclear cell surfaces. Cellular Fc receptors failed to recognize the Fc region of aggregated IgG that had been blocked by SPA. Moreover, fluoresceinated SPA that had been complexed either with anti-Fab (B-cell specific) or T cell-specific antisera prevented the nonspecific binding of these reagents to the IgG-Fc receptors on mononuclear cells, thereby permitting the latter to be properly identified as B or T lymphocytes. In addition, when unconjugated SPA was added to presensitized target cells in a test for antibody-dependent cell-mediated cytotoxicity, cytolysis was abrogated.

    Title Comparative Evaluation of Commercial Precipitating Antisera Against Human Iga.
    Date June 1971
    Journal The Journal of Laboratory and Clinical Medicine
    Title A New 'total' Activin B Enzyme-linked Immunosorbent Assay (elisa): Development and Validation for Human Samples.
    Date
    Journal Clinical Endocrinology
    Excerpt

    There are currently no sensitive and specific assays for activin B that could be utilized to study human biological fluids. The aim of this project was to develop and validate a 'total' activin B ELISA for use with human biological fluids and establish concentrations of activin B in the circulation and fluids from the reproductive organs.

    Title Myoelectric and Mechanical Changes Elicited by Ischemic Preconditioning in the Feline Hindlimb.
    Date
    Journal Journal of Electromyography and Kinesiology : Official Journal of the International Society of Electrophysiological Kinesiology
    Excerpt

    Tourniquet use is fraught with potential complications. For example, ischemia produced by the tourniquet may lead to nerve and muscle injuries. One technique shown in cardiovascular and free-flap surgery to improve the viability of muscle subjected to ischemia is preconditioning. This technique involves an initial brief period of ischemia, followed by reperfusion before a prolonged ischemic episode. The purpose of this study was to explore ischemic preconditioning as a method to reduce tourniquet-related morbidity. In six cats, one leg was preconditioned by 10 min of tourniquet-induced ischemia followed by 10 min of reperfusion. The contralateral limb was not preconditioned. Both limbs underwent 1 h of tourniquet inflation followed by a 2-h recovery period. Isometric force and electromyographic (EMG) amplitude were recorded throughout the procedure at 20-min intervals in both medial gastrocnemius muscles. Analysis of variance (ANOVA) with repeated measures shows that, after 60 min of tourniquet application, maximal isometric force was significantly larger in the preconditioned group. Furthermore, the EMG amplitude during recovery was found to be significantly larger in the preconditioned limbs. These results suggest that preconditioning improves skeletal muscle viability in vivo. Further research is needed, however, to assess the long-term effects of this technique, and to delineate appropriate preconditioning protocols that would improve surgical outcome without significantly increasing the complexity of the procedures.

    Title Developmental Programming: Impact of Excess Prenatal Testosterone on Intrauterine Fetal Endocrine Milieu and Growth in Sheep.
    Date
    Journal Biology of Reproduction
    Excerpt

    Prenatal testosterone excess in sheep leads to reproductive and metabolic disruptions that mimic those seen in women with polycystic ovary syndrome. Comparison of prenatal testosterone-treated sheep with prenatal dihydrotestosterone-treated sheep suggests facilitation of defects by androgenic as well as androgen-independent effects of testosterone. We hypothesized that the disruptive impact of prenatal testosterone on adult pathology may partially depend on its conversion to estrogen and consequent changes in maternal and fetal endocrine environments. Pregnant Suffolk sheep were administered either cottonseed oil (control) or testosterone propionate in cottonseed oil (100 mg, i.m. twice weekly), from Day 30 to Day 90 of gestation (term is ~147 d). Maternal (uterine) and fetal (umbilical) arterial samples were collected at Days 64-66, 87-90, and 139-140 (range; referred to as D65, D90, and D140, respectively) of gestation. Concentrations of gonadal and metabolic hormones, as well as differentiation factors, were measured using liquid chromatography/mass spectrometer, radioimmunoassay, or ELISA. Findings indicate that testosterone treatment produced maternal and fetal testosterone levels comparable to adult males and D65 control male fetuses, respectively. Testosterone treatment increased fetal estradiol and estrone levels during the treatment period in both sexes, supportive of placental aromatization of testosterone. These steroidal changes were followed by a reduction in maternal estradiol levels at term, a reduction in activin A availability, and induction of intrauterine growth restriction in D140 female fetuses. Overall, our findings provide the first direct evidence in support of the potential for both androgenic as well as estrogenic contribution in the development of adult reproductive and metabolic pathology in prenatal testosterone-treated sheep.

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