Biomedical subjects

J A Thompson

Publications and source records attributed to J A Thompson.

At least 37 records · Page 2Linked to original sources

PubMed2001

Ranch-management factors associated with antibody seropositivity for Neospora caninum in consignments of beef calves in Texas, USA.

A study was conducted with a 1998 retained-ownership population of Texas (USA) beef calves to determine the ranch-management practices associated with calf seroprevalence to Neospora caninum. Management practices of 76 Texas ranches that consigned 760 calves to a retained-ownership feedlot program were reviewed from a mailed questionnaire. Ninety-nine of 760 (13%; 95% CI, 9.4%, 17.7%) calves were positive to N. caninum and 59% of the ranches consigned at least one positive calf. In the logistic multiple-regression model which controlled for overdispersion, increased odds of calf-level seropositivity was associated with seasonal calving patterns, with stocking>1cow/calfunit/2.2ha, using a round-bale feeder, allowing wildlife access to the weaning supplement, and self-reared replacement heifers. However, decreased odds of seropositivity was associated with using a cattle-working dog and with using a self-contained cattle feeder. There was substantial overdispersion due to ranch.

Animal Husbandry↗

PubMed2001

Association of serologic status for Neospora caninum and postweaning feed efficiency in beef steers.

OBJECTIVE: To determine the effects of serologic status for Neospora caninum on short-term weight gain, feed intake, and feed efficiency (feed intake/gain). DESIGN: Longitudinal observational study. ANIMALS: 34 weaned mixed-breed beef steers. PROCEDURE: Serologic status for N. caninum was determined for each steer on days 0 (weaning), 88, 116, 144, 172, and 200, using an agglutination test. Individual steer body weight was measured on days 0, 88, 116, 144, 172, 200, and 242 (slaughter). Daily feed intake was monitored from days 116 through 242. Serologic status was matched to animal performance for the period immediately following serum sample collection. A mixed mode, using repeated-measures with an unstructured covariance matrix, was used in the analysis. Breed, age, and pen effects were controlled for in the analysis. RESULTS: A reduction in average daily gain for the period following a positive serologic result was detected for the entire trial (6 measurements/steer). This may have been attributed to a significant impairment in feed efficiency rather than to an impairment in feed intake. Changes in serologic status in individual steers over time were common; additionally, the effects of serologic status on steer performance were also transitory. CONCLUSIONS AND CLINICAL RELEVANCE: Significant reductions in short-term weight gain and feed efficiency were associated with the presence of antibodies against N. caninum in postweaning beef steers.

Agglutination Tests↗

PubMed2001

Comparisons of hydroperoxide isomerase and monooxygenase activities of cytochrome P450 for conversions of allylic hydroperoxides and alcohols to epoxyalcohols and diols: probing substrate reorientation in the active site.

Ferric cytochrome P450 (P450) catalyzes intramolecular rearrangements of hydroperoxides to diols by heterolytic scission of the O-O bond and insertion of the terminal oxygen atom into the resulting alcohol. The goals of this work were to further characterize the regio- and stereochemistry of P450 isomerase activity using allylic hydroperoxides and to compare these reactions with NADPH-supported monooxygenations of the corresponding alcohols. Microsomes from phenobarbital-treated rats or purified P450 2B1 catalyzed the conversions of several peroxyquinols, including 2-tert-butyl-4-hydroperoxy-4-methylcyclohexa-2,5-dien-1-one (BMPOOH) and its 2,6-dimethyl derivative (TMPOOH), to diols and to alpha,beta-epoxyquinols by predominant (>93%) cis addition of oxygen to the least hindered double bond. Monooxygenation of the 4-hydroxy analogues (quinols) yielded identical cis-epoxyquinols, and both isomerization and monooxygenation reactions exhibited similar enantioselectivities (32-74% enantiomer excess). Regioselectivities were similar for BMPOOH and BMPOH (epoxyquinol:diol ratios of 5.8-7.6), but quite different for TMPOOH and TMPOH (ratios of 0.6 and 6.4, respectively). Bulky peroxyquinols and quinols derived from the A-rings of 17beta-estradiol and estrone were utilized to further compare these reactions. Both estradiol derivatives underwent approximately equal amounts of 6beta-hydroxylation and 1,2-epoxidation. The estrone derivatives also underwent 6beta-hydroxylation, but only estrone quinol yielded a second product consistent with hydroxylation at position 16. The results support several conclusions. (i) Allylic hydroperoxides may be isomerized to alpha,beta-epoxyalcohols by a heterolytic O-O cleavage mechanism with high stereoselectivity. (ii) Hydroperoxide isomerization is an efficient process relative to monooxygenation. (iii) Isomerase substrates remain in proximity to the P450 oxoferryl intermediate and are rapidly captured by the oxidant. Monooxygenase substrates, on the other hand, may bind to ferric P450 in multiple orientations and undergo more extensive substrate reorientation prior to oxidative attack.

Animals↗

PubMed2001

Prognostic factors analysis of 17,600 melanoma patients: validation of the American Joint Committee on Cancer melanoma staging system.

PURPOSE: The American Joint Committee on Cancer (AJCC) recently proposed major revisions of the tumor-node-metastases (TNM) categories and stage groupings for cutaneous melanoma. Thirteen cancer centers and cancer cooperative groups contributed staging and survival data from a total of 30,450 melanoma patients from their databases in order to validate this staging proposal. PATIENTS AND METHODS: There were 17,600 melanoma patients with complete clinical, pathologic, and follow-up information. Factors predicting melanoma-specific survival rates were analyzed using the Cox proportional hazards regression model. Follow-up survival data for 5 years or longer were available for 73% of the patients. RESULTS: This analysis demonstrated that (1) in the T category, tumor thickness and ulceration were the most powerful predictors of survival, and the level of invasion had a significant impact only within the subgroup of thin (< or = 1 mm) melanomas; (2) in the N category, the following three independent factors were identified: the number of metastatic nodes, whether nodal metastases were clinically occult or clinically apparent, and the presence or absence of primary tumor ulceration; and (3) in the M category, nonvisceral metastases was associated with a better survival compared with visceral metastases. A marked diversity in the natural history of pathologic stage III melanoma was demonstrated by five-fold differences in 5-year survival rates for defined subgroups. This analysis also demonstrated that large and complex data sets could be used effectively to examine prognosis and survival outcome in melanoma patients. CONCLUSION: The results of this evidence-based methodology were incorporated into the AJCC melanoma staging as described in the companion publication.

Adolescent↗

PubMed2001

Final version of the American Joint Committee on Cancer staging system for cutaneous melanoma.

PURPOSE: To revise the staging system for cutaneous melanoma under the auspices of the American Joint Committee on Cancer (AJCC). MATERIALS AND METHODS: The prognostic factors analysis described in the companion publication (this issue), as well as evidence from the published literature, was used to assemble the tumor-node-metastasis criteria and stage grouping for the melanoma staging system. RESULTS: Major changes include (1) melanoma thickness and ulceration but not level of invasion to be used in the T category (except for T1 melanomas); (2) the number of metastatic lymph nodes rather than their gross dimensions and the delineation of clinically occult (ie, microscopic) versus clinically apparent (ie, macroscopic) nodal metastases to be used in the N category; (3) the site of distant metastases and the presence of elevated serum lactic dehydrogenase to be used in the M category; (4) an upstaging of all patients with stage I, II, and III disease when a primary melanoma is ulcerated; (5) a merging of satellite metastases around a primary melanoma and in-transit metastases into a single staging entity that is grouped into stage III disease; and (6) a new convention for defining clinical and pathologic staging so as to take into account the staging information gained from intraoperative lymphatic mapping and sentinel node biopsy. CONCLUSION: This revision will become official with publication of the sixth edition of the AJCC Cancer Staging Manual in the year 2002.

Humans↗

PubMed2001

Acidic fibroblast growth factor (FGF-1) signaling inhibits peroxynitrite-induced cell death during pancreatic tumorigenesis.

Previous immunohistochemical studies have demonstrated enhanced appearance of FGF-1 and nitrotyrosine, a footprint of reactive nitrogen species peroxynitrite (ONOO(-)), in human pancreatic adenocarcinoma. We have examined the consequences of constitutive exposure to FGF-1 in nontumorigenic rat ductal epithelial cells (ARIP). ARIP cells were transduced with either a secreted chimera of FGF-1, ARIP(FGF-1), or a control plasmid, 65 RIP(betag). These cells were evaluated for alteration in growth and morphology, responses to ONOO(-) (protein tyrosine nitration/phosphorylation), and in vivo tumor formation. ARIP(FGF-1) cells, in contrast to 65 RIP(betag), demonstrated a transformed morphology, a 2-fold increased growth rate, and enhanced protein tyrosine phosphorylation. Treatment with 150 microM ONOO(-) resulted in 86 and 7% (p <.01) death of ARIP(betag) and ARIP(FGF-1), respectively. Exposure of 65 RIP(betag) cells to ONOO(-) enhanced tyrosine phosphorylation and tyrosine nitration of several polypeptides. Cell signaling by FGF-1 enhanced both phosphorylation and nitration of tyrosine residues in target proteins modified by ONOO(-). ARIP(betag) cells failed to exhibit tumor formation in nude mice, but at d 7 in vivo cells were TUNEL and nitrotyrosine positive and FGF-1 negative. ARIP(FGF-1) cells readily formed tumor nodules, exhibiting features of pancreatic adenocarcinoma and demonstrating FGF-1-positive, nitrotyrosine-positive, and TUNEL-negative epithelium. These results suggest an interdependent role between FGF-1 and ONOO(-) during the development and progression of pancreatic adenocarcinoma.

Animals↗

PubMed2001

Studies using structural analogs and inbred strain differences to support a role for quinone methide metabolites of butylated hydroxytoluene (BHT) in mouse lung tumor promotion.

Chronic treatment of BALB and GRS mice with BHT (2,6-di-tert-butyl-4-methylphenol) following a single urethane injection increases lung tumor multiplicity, but this does not occur in CXB4 mice. Previous data suggest that promotion requires the conversion of BHT to a tert-butyl-hydroxylated metabolite (BHTOH) in lung and the subsequent oxidation of this species to an electrophilic quinone methide. To obtain additional evidence for the importance of quinone methide formation, structural analogs that form less reactive quinone methides were tested and found to lack promoting activity in BHT-responsive mice. The possibility that promotion-unresponsive strains are unable to form BHTOH was tested by substituting this compound for BHT in the promotion protocol using CXB4 mice. No promotion occurred, and in-vitro work demonstrated that CXB4 mice are, in fact, capable of producing BHTOH and its quinone methide, albeit in smaller quantities. Incubations with BALB lung microsomes and radiolabeled substrates confirmed that more covalent binding to protein occurs with BHTOH than with BHT and, in addition, BHTOH quinone methide is considerably more toxic to mouse lung epithelial cells than BHT quinone methide. These data are consistent with the hypothesis that a two-step oxidation process, i.e. hydroxylation and quinone methide formation, is required for the promotion of mouse lung tumors by BHT.

Animals↗

PubMed2001

Development of a measure of attitude toward nutrition in patient care.

BACKGROUND: Development of reliable measures of medical student and resident attitudes about nutrition in patient care is needed before the effects of educational interventions or clinical experience can be gauged. This report describes the systematic development of a measure of attitude toward nutrition in patient care. It presents evidence about scale reliability and the absence of response bias that endorses the trustworthiness of data from the measure. METHODS: An eight-step attitude scale development procedure was used to create the Nutrition In Patient care Survey (NIPS). Data from five samples of first- and second-year medical students and first-year medical residents were subjected to factor analysis (PA2, varimax rotation), reliability analyses, and statistical analyses to test for demographic bias in the attitude data. RESULTS: A 45-item attitude measure was developed that contains five subscales derived from the factor analysis: (1) nutrition in routine care (NRC, 8 items); (2) clinical behavior (CB, 20 items); (3) physician-patient relationship (PPR, 8 items); (4) patient behavior/motivation (PBM, 3 items); and (5) physician efficacy (PE, 6 items). Each subscale yields reliable data in terms of internal consistency (alpha coefficients) and stability (test-retest reliability). Medical student and resident demographic variables have negligible influence on attitude scores. DISCUSSION: The NIPS subscales yield reliable data that can be used to assess outcomes in evaluation research on educational or clinical interventions or to predict patient care practices. Systematic attitude scale development increases the likelihood that the resulting measures will produce useful, trustworthy data.

Adult↗

PubMed2001

Radioimaging of implants in rats using Tc-99m-MDP.

Radiopharmaceutical isotopes are widely used clinically to detect tumors of osteogenic origin. One example is Technetium-99m methylene diphosphonate (Tc-99m-MDP). When viewed with a gamma camera, the concentration of the isotope (increased gamma activity) indicates an area of increased bone activity. This technology provides an opportunity to measure bone growth around implants in vivo. The purpose of this study was to measure Tc-99m-MDP activity around titanium alloy implants placed in the tibiae of rats. Some implant sites were treated with a growth factor; other sites served as controls. The hypothesis tested was that implants placed with a growth factor would have greater associated Tc-99m-MDP activity. Twelve adult male Sprague-Dawley rats were anesthetized and surgical access to the medial proximal tibiae was obtained. Titanium alloy screw implants were placed in six animals along with 65 microgram of acidic fibroblast growth factor (FGF-1); the other six animals received implants only and served as controls. After five days, rats were injected with 1500 microCi of Tc-99m-MDP. After 3 hours, rats were imaged with a gamma camera. The Tc-99m-MDP intensity associated with each implant was quantified and the means for each group were compared using ANOVA. Implants treated with FGF-1 demonstrated significantly more Tc-99m-MDP activity than implants alone. This suggests that Tc-99m-MDP analysis may be a useful tool for determining bone growth around implants in laboratory animals in vivo.

Analysis of Variance↗

PubMed2001

Increased dietary salt accelerates chronic allograft nephropathy in rats.

BACKGROUND: Chronic allograft nephropathy (CAN), a major problem in renal transplantation, is related to both alloantigen-dependent and -independent processes. Because dietary salt intake modulated glomerular production of transforming growth factor-beta, which has been shown to play an important role in CAN, we hypothesized that dietary salt would directly enhance renal injury in a rodent model of CAN. METHODS: Dietary NaCl was increased from 1.0% (normal) to 8.0% in a group of Fisher/Lewis rats 25 days following orthotopic renal transplantation and was continued until 16 weeks after transplantation. RESULTS: Blood pressure, which was recorded using radiotelemetry in the first eight-weeks post-transplantation, did not differ between the groups, but allograft recipients on the 8.0% NaCl diet rapidly demonstrated increased urinary albumin excretion. Renal function determined by dynamic functional imaging was worse in allograft recipients on the 8.0% NaCl diet by six weeks following transplantation. Histologic examination at 16 weeks confirmed a significant increase in allograft damage in the 8.0% NaCl group compared with allografts from rats on 1.0% NaCl diet. These findings included glomerulosclerosis and tubulointerstitial injury that consisted of fibrosis, tubular atrophy and dilation, intratubular casts, and tubular epithelial cell damage. Small arteries and arterioles did not show evidence of damage from hypertension or other abnormality. CONCLUSIONS: In this model of CAN, renal allograft dysfunction preceded hypertension and was accelerated significantly by an increase in dietary salt.

Albuminuria↗

PubMed2001

Randomized multicenter phase II trial of subcutaneous recombinant human interleukin-12 versus interferon-alpha 2a for patients with advanced renal cell carcinoma.

Recombinant human interleukin-12 (rHuIL-12) is a pleiotropic cytokine with anticancer activity against renal cell carcinoma (RCC) in preclinical models and in a phase I trial. A randomized phase II study of rHuIL-12 compared with interferon-alpha (IFN-alpha) evaluated clinical response for patients with previously untreated, advanced RCC. Patients were randomly assigned 2:1 to receive either rHuIL-12 or IFN-alpha2a. rHuIL-12 was administered by subcutaneous (s.c.) injection on days 1, 8, and 15 of each 28-day cycle. The dose of IL-12 was escalated during cycle 1 to a maintenance dose of 1.25 microg/kg. IFN was administered at 9 million units by s.c. injection three times per week. Serum concentrations of IL-12, IFN-gamma, IL-10, and neopterin were obtained in 10 patients treated with rHuIL-12 after the first full dose of 1.25 microg/kg given on day 15 (dose 3) of cycle 1 and again after multiple doses on day 15 (dose 6) of cycle 2. Thirty patients were treated with rHuIL-12, and 16 patients were treated with IFN-alpha. Two (7%) of 30 patients treated with rHuIL-12 achieved a partial response, and the trial was closed to accrual based on the low response proportion. IL-12 was absorbed rapidly after s.c. drug administration, with the peak serum concentration appearing at approximately 12 h in both cycles. Serum IL-12 concentrations remained stable on multiple dosing. Levels of IFN-gamma, IL-10, and neopterin increased with rHuIL-12 and were maintained in cycle 2. rHuIL-12 is a novel cytokine with unique pharmacologic and pharmacodynamic features under study for the treatment of malignancy and other medical conditions. The low response proportion associated with rHuIL-12 single-agent therapy against metastatic RCC was disappointing, given the preclinical data. Further study of rHuIL-12 for other medical conditions is underway. For RCC, the study of new cytokines is of the highest priority.

Adult↗

PubMed2001

Mares with delayed uterine clearance have an intrinsic defect in myometrial function.

Persistent, postmating endometritis affects approximately 15% of mares and results in reduced fertility and sizable economic losses to the horse-breeding industry. Mares that are susceptible to postmating endometritis have delayed uterine clearance associated with reduced uterine contractility. Unfortunately, the mechanism for reduced uterine contractility remains an enigma. The present study examined the hypothesis that mares with delayed uterine clearance have an intrinsic contractile defect of the myometrium. Myometrial contractility was evaluated in vitro by measuring isometric tension generated by longitudinal and circular uterine muscle strips in response to KCl, oxytocin, and prostaglandin F(2alpha) (PGF(2alpha)) for young nulliparous mares, older reproductively normal mares, and older mares with delayed uterine clearance. In addition, intracellular Ca(2+) regulation was evaluated using laser cytometry to measure oxytocin-stimulated intracellular Ca(2+) transients of myometrial cells loaded with a Ca(2+)-sensitive fluorescent dye, fluo-4. For all contractile agonists, myometrium from mares with delayed uterine clearance failed to generate as much tension as myometrium from older normal mares. Oxytocin-stimulated intracellular Ca(2+) transients were similar for myometrial cells from mares with delayed uterine clearance and from older normal mares, suggesting that the contractile defect did not result from altered regulation of intracellular Ca(2+) concentration. Furthermore, no apparent age-dependent decline was observed in myometrial contractility; KCl-depolarized and oxytocin-stimulated longitudinal myometrium from young normal mares and older normal mares generated similar responses. However, circular myometrium from young normal mares failed to generate as much tension as myometrium from older normal mares when stimulated with oxytocin or PGF(2alpha), suggesting possible age-related alterations in receptor-second messenger signaling mechanisms downstream of intracellular Ca(2+) release. In summary, for mares with delayed uterine clearance, an intrinsic contractile defect of the myometrium may contribute to reduced uterine contractility following breeding.

Aniline Compounds↗

PubMed2001

America's best medical schools: a critique of the U.S. News & World Report rankings.

Rankings of American medical schools published annually by the news magazine U.S. News & World Report are widely used to judge the quality of the schools and their programs. The authors describe and then critique the rankings on methodologic and conceptual grounds, arguing that the annual U.S. News medical school evaluation falls short in both areas. Three categories of program quality indicators different from those used by U.S. News are presented as alternative ways to judge medical schools. The authors conclude that the annual U.S. News & World Report rankings of American medical schools are ill-conceived; are unscientific; are conducted poorly; ignore medical school accreditation; judge medical school quality from a narrow, elitist perspective; and do not consider social and professional outcomes in program quality calculations. The medical school rankings have no practical value and fail to meet standards of journalistic ethics.

Bias↗

PubMed2001

Acidic fibroblast growth factor attenuates the cytotoxic effects of peroxynitrite in primary human osteoblast precursors.

BACKGROUND: Skeletal injury and associated ischemia and inflammation induce the generation of pro-oxidants such as peroxynitrite (ONOO-), which has been demonstrated to induce apoptosis in several cell lines. Fibroblast growth factor (FGF-1) is important for coordinating osteogenesis and angiogenesis of osseous repair. In vitro studies were performed examining the effect of FGF-1 on human osteoblast progenitor stromal stem (HSS) cell proliferation, differentiation, and response to ONOO-. METHODS: HSS cells were isolated and growth kinetics determined in the presence and absence of FGF-1. The effect of FGF-1 on HSS cell expression of osteoblast-specific osteopontin and osteocalcin mRNA and protein was examined by reverse transcriptase polymerase chain reaction and Western blot techniques. To determine the sensitivity of HSS cells to ONOO- in the absence and presence of FGF-1 pretreatment, cells were exposed to varying concentrations of the oxidant and examined for cell death using quantitative fluorescence staining with fluorescein diacetate and propidium diacetate. RESULTS: Treatment of HSS cells with FGF-1 significantly enhanced cellular growth rates by 5 days (4.6 x 105 cells/mL vs. 3.1 x 105 cells/mL) and induced expression of both osteopontin and osteocalcin mRNA and protein. Exposure of HSS cells to ONOO- resulted in a dose- and time-dependent delayed cell death that was more characteristic of apoptosis than necrosis. Pretreatment of HSS cells with FGF-1 prevented ONOO- mediated apoptosis. CONCLUSION: In vitro, treatment of HSS cells with FGF-1 stimulates cell growth and induces expression of differentiation markers specific to osteoblasts. FGF-1 treatment renders osteoblast precursors resistant to the cytotoxic effects of ONOO-. These results suggest that FGF-1 promotes the progression of bone repair mechanisms by increasing the population of osteoblasts and imparting protection to the cell line from the hostile inflammatory environment.

Apoptosis↗

PubMed2001

Tyrosine kinase receptor activation inhibits NPR-C in lung arterial smooth muscle cells.

We have previously demonstrated that expression of the atrial natriuretic peptide (ANP) clearance receptor (NPR-C) is reduced selectively in the lung of rats and mice exposed to hypoxia but not in pulmonary arterial smooth muscle cells (PASMCs) cultured under hypoxic conditions. The current study tested the hypothesis that hypoxia-responsive growth factors, fibroblast growth factors (FGF-1 and FGF-2) and platelet-derived growth factor-BB (PDGF-BB), that activate tyrosine kinase receptors can reduce expression of NPR-C in PASMCs independent of environmental oxygen tension. Growth-arrested rat PASMCs were incubated under hypoxic conditions (1% O2) for 24 h; with FGF-1, FGF-2, or PDGF-BB (0.1-20 ng/ml for 1-24 h); or with ANG II (1-100 nM), endothelin-1 (ET-1, 0.1 microM), ANP (0.1 microM), sodium nitroprusside (SNP, 0.1 microM), or 8-bromo-cGMP (0.1 mM) for 24 h under normoxic conditions. Steady-state NPR-C mRNA levels were assessed by Northern blot analysis. FGF-1, FGF-2, and PDGF-BB induced dose- and time-dependent reduction of NPR-C mRNA expression within 1 h at a threshold concentration of 1 ng/ml; hypoxia, ANG II, ET-1, ANP, SNP, or cGMP did not decrease NPR-C mRNA levels in PASMCs under the above conditions. Downregulation of NPR-C expression by FGF-1, FGF-2, and PDGF-BB was inhibited by the selective FGF-1 receptor tyrosine kinase inhibitor PD-166866 and mitogen-activated protein/extracellular signal-regulated kinase inhibitors U-0126 and PD-98059. These results indicate that activation of tyrosine kinase receptors by hypoxia-responsive growth factors, but neither hypoxia per se nor activation of G protein-coupled receptors, inhibits NPR-C gene expression in PASMCs. These results suggest that FGF-1, FGF-2, and PDGF-BB play a role in the signal transduction pathway linking hypoxia to altered NPR-C expression in lung.

Animals↗

PubMed2001

Promoting collaborative teaching in clinical education.

BACKGROUND: Although the practice of medicine is increasingly a multidisciplinary effort, clinical teaching of medical students is accomplished primarily within a departmental structure. PURPOSE: The purpose of this study was to identify subject matter within the clinical curriculum that could serve as focus for multidisciplinary teaching. METHODS: A questionnaire was sent to 13 clerkship directors (representing required clerkships) at a large Midwestern medical school in which they were asked to rate a list of 631 patient problems as critical (primary), important (secondary), or "nice to know," relevant to their respective clerkship objectives. RESULTS: All clerkship directors completed the questionnaire. There were 523 items that were considered primary, and over 90% of these items were listed as either primary or secondary in more than 1 clerkship. Twelve topics were considered primary or secondary by at least 5 clerkship directors. Four clerkship directors identified 43 patient problems, and 3 clerkship directors identified 92 topics as primary or secondary clerkship objectives. CONCLUSIONS: In this study, listing of patient problems across clerkships demonstrated significant overlap of the clinical curriculum, suggesting multiple opportunities for faculty collaboration in clinical education.

Clinical Clerkship↗

PubMed2001

Acidic fibroblast growth factor signaling inhibits peroxynitrite-induced death of osteoblasts and osteoblast precursors.

After trauma injury to the musculoskeletal system, conditions such as ischemia and inflammation involve excess production of superoxide (O2*), nitric oxide (*NO), and their reaction product, peroxynitrite (ONOO-). Exposure of murine osteoblasts and rat-derived primary osteoblast precursors to ONOO- resulted in a dose- and time-dependent delayed cell death that was more characteristic of apoptosis than necrosis. Exposure of both cell populations to ONOO- immediately enhanced phosphorylation and nitration of tyrosine residues within several polypeptides. Treatment of osteoblasts and osteoblast precursors with exogenous acidic fibroblast growth factor (FGF-1) enhanced cellular growth, increased endogenous levels of tyrosine phosphorylation, and significantly induced expression of both osteopontin and osteocalcin messenger RNA (mRNA) as well as osteopontin protein. Pretreatment of both cell populations with exogenous FGF-1 prevented ONOO(-)-mediated death. Cell signaling induced by FGF-1 pretreatment had no major effect of total levels of tyrosine nitration after ONOO- treatment. Collectively, these in vitro efforts show that FGF-1 signaling renders osteoblasts and osteoblast precursors resistant to the cytotoxic effects of ONOO-. Consequently, results presented here predict the therapeutic use of this growth factor for promoting the progression of bone repair mechanisms after fracture trauma.

Animals↗

PubMed2001

Activation of cumulus-free equine oocytes: effect of maturation medium, calcium ionophore concentration and duration of cycloheximide exposure.

Two different culture media (TCM-199 and follicular fluid), two activation treatments (10 and 50 micromol calcium ionophore l(-1)) and three culture periods with cycloheximide were evaluated to find effective culture conditions for activation of cumulus-free equine oocytes. Oocytes were collected by scraping the follicle walls of ovaries obtained from an abattoir. Oocytes with expanded cumuli were matured at 38.2 degrees C in a humidified atmosphere of 5% CO(2) in air, in either TCM-199 with 10% fetal bovine serum (FBS) and 5 microU FSH ml(-1), or in 100% follicular fluid derived from a preovulatory follicle 24 h after injection of hCG. After 40--42 h of in vitro maturation, oocytes were denuded by gentle pipetting in TCM-199 plus 10% FBS with hyaluronidase. Oocytes with intact cytoplasmic membranes (n = 398; 94% presumed metaphase II) were treated in protein-free PBS with 10 or 50 micromol calcium ionophore l(-1) for 5 min. After washing, the oocytes were cultured in TCM-199 containing 10% FBS and 10 microg cycloheximide ml(-1) for 6 h, in cycloheximide for 6 h and then in cycloheximide-free medium for 18 h, or in cycloheximide for 24 h. The oocytes were fixed and evaluated by fluorescence microscopy. Oocytes with pronucleus I--II (dense to decondensing chromatin), pronucleus III--IV (decondensed chromatin) or progressing towards the first cleavage division were considered activated. The activation rate for oocytes matured in TCM-199 was significantly (P < 0.05) higher than for oocytes matured in follicular fluid (49% (99/204) versus 35% (60/171), respectively; P < 0.05). Culture with cycloheximide for 24 h resulted in a significantly higher rate of activation (67%, 74/111) than did the 6 h (33%, 44/136) or 6 h plus 18 h (32%, 41/128) treatments. The highest rate of activation (82%) was observed in oocytes matured in TCM-199, treated with 50 micromol calcium ionophore l(-1) and cultured with cycloheximide for 24 h.

Animals↗