Clinical Chemistry Profiles Research Articles

Aetiology and Prognostic Implication of Severe Jaundice in Surgical Trauma Patients

Background: Pronounced postoperative jaundice occurs not infrequently in trauma patients. The aim of this study was to elucidate the implication of early, pronounced jaundice (serum-bilirubin >100 μmol·1̄¹) for 30-day survival of such patients. Methods: From 1995 through 2001, 53 surgical trauma patients developing pronounced postoperative jaundice were identified. Nine were excluded from the study because of major hepatobiliary injury or pre-existing liver disease. The clinical course and laboratory chemistry profiles of the remaining 44 patients were analysed. Results: Thirty-one patients survived and 13 died within 30 days of trauma. Non-survivors had higher age, higher injury severity score (ISS) and lower probability of survival (PS) (P ≺ 0.05) than survivors. ISS averaged 34 in survivors and 45 in non-survivors. Survivors and non-survivors received a mean of 46 (range 10-97) and 55 units of blood (range 11-128), respectively (P = 0.366). Systematic hypotension, local infections and sepsis were common in both groups. Bilirubin levels peaked around the 11th day in survivors (median 189 μmol·1̄¹). In non-survivors, serum bilirubin values rose progressively, reaching maximum levels at time of death (median 231 μmol·1̄¹). These patients died in a setting of sepsis and multiple organ failure. Conclusion: Large endogenous production of bilirubin because of rapid breakdown of transfused and extravasated blood can cause pronounced jaundice in multitransfused trauma patients. In such patients, serum bilirubin rising > 100 μmol·1̄¹ does not by itself signal poor outcome. However, progressive pronounced jaundice outlasting the trauma incident by 10-12 days portends fatal outcome for the patient.

High blood glutathione levels accompany excellent physical and mental health in women ages 60 to 103 years

Earlier we found a high percentage of subnormal total glutathione (GT) levels in blood from elderly subjects and patients with chronic diseases. These findings suggested a hypothesis that high levels of GT in the blood occur in old persons who are in excellent physical and mental health. To this end, we recruited 87 white women who ranged in age from 60 to 103 years and reported that they felt healthy. Their health was verified with physical examinations, clinical chemistry profiles, psychosocial assessments, and blood GT determinations. This evaluation was performed in three waves over a 5-year period. The values were compared with those from representative individuals in this region and with normal national data. The results verified that these healthy subjects were in top physical and mental health. We also found that subjects of all ages had very high blood GT levels in waves I and II but only normal levels in wave III. These findings confirm that high blood GT concentrations and excellent physical and mental health are characteristics of long-lived women. (J Lab Clin Med 2002;140:413-7)

A 13-week subchronic intravaginal toxicity study of the novel broad-spectrum anti-HIV and spermicidal agent, N-[2-(1-cyclohexenyl)ethyl]-N'-[2-(5-bromopyridyl)]-thiourea (PHI-346) in mice.

The nonnucleoside inhibitor (NNI) of HIV-1 reverse transcriptase, PHI-346 (N-[2-(1-cyclohexenyl)ethyl]-N'-[2-(5-bromopyridyl)]-thiourea), is a dual-function spermicidal agent with potent anti-HIV activity against drug-sensitive as well as drug-resistant HIV-1 strains with genotypic and phenotypic NNI resistance. PHI-346 was formulated via a lipophilic gel-microemulsion for intravaginal use as a potential dual-function microbicide. To evaluate the toxicity potential of short-term intravaginal exposure to PHI-346, groups of 15 female B6C3F1 and CD-1 mice were exposed intravaginally to a gel-microemulsion containing 0, 0.5, 1.0, or 2.0% PHI-346, 5 days per week for 13 consecutive weeks. On a molar basis, these concentrations of PHI-346 are 350 to 1,400-times higher than its spermicidal EC50 and nearly 5 x 10(6) to 2 x 10(7) times higher than its in vitro anti-HIV IC50. After 13 weeks of intravaginal treatment, B6C3F1 mice were evaluated for survival, body weight gain, absolute and relative organ weights. Blood was analyzed for hematology and clinical chemistry profiles. Microscopic examination was performed on hematoxylin and eosin-stained tissue sections from each study animal. Placebo control and PHI-346 dosed female CD-1 mice were mated with untreated males in order to evaluate if PHI-346 has any deleterious effects on the reproductive performance. There were no treatment-related mortalities. Mean body weight gain during the dosing period was not reduced by PHI-346 treatment. The hemogram or blood chemistry profiles revealed lack of systemic toxicity following daily intravaginal instillation of PHI-346 for 13 weeks. No clinically significant changes in absolute and relative organ weights were noted in PHI-346 dose groups. Extensive histopathological examination of tissues revealed no treatment-related abnormalities in any of the three PHI-346 dose groups. Repeated intravaginal exposure of CD-1 mice to increasing concentrations of PHI-346 for 13 weeks had no adverse effect on their subsequent reproductive capability, perinatal outcome, growth, and development of offspring. Collectively, these findings demonstrate that repetitive intravaginal administration of PHI-346 at concentrations as high as 1,400-times its spermicidal EC50 and 2 x 10(7) times its in vitro anti-HIV IC50 was not associated with local or systemic toxicity and did not adversely affect the reproductive performance in mice. PHI-346 may be useful as an active ingredient of a safe vaginal microbicide for prevention of the sexual transmission of multidrug-resistant HIV-1.

Two-year toxicity and carcinogenicity studies in B 6C 3F 1 mice with 5-bromo-6-methoxy-5,6-dihydro-3′-azidothymidine-5′-( p-bromophenyl) methoxyalaninyl phosphate (WHI-07), a novel anti-HIV and contraceptive agent

The zidovudine derivative, 5-bromo-6-methoxy-5,6-dihydro-3′-azidothymidine-5′-( p-bromophenyl) methoxy alaninyl phosphate (WHI-07), is a dual-function spermicidal and anti-HIV agent with contraceptive and microbicidal activity. In previous two subchronic toxicity studies, intravaginal application of 0.5–2.0% WHI-07, for up to 13 weeks, was shown to cause no local, systemic or reproductive toxicity. To evaluate the toxicity and carcinogenic potential of long-term exposure to WHl-07, groups of 50 female B 6C 3F 1 mice were given no treatment or exposed intravaginally to a gel-microemulsion formulation with and without 2.0% WHI-07, 5 days per week for 2 years. The dose of WHI-07 was equivalent to 5700 times its spermicidal EC 50 and 5.7×10 6 times its anti-HIV IC 50. The endpoints that were evaluated included survival, body weight, hematologic and clinical chemistry profiles, absolute and relative organ weights, and histopathology. No significant differences in mean body weight gain and survival were found among the groups of untreated control, placebo control, and 2% WHI-07-treated mice at the end of the 2-year study. The hematological and clinical chemistry profiles did not reveal any toxicologically significant changes that could be attributed to WHI-07 treatment. No clinically significant changes in absolute and relative organ weights were noted in the WHI-07 group. A variety of neoplastic and nonneoplastic lesions which were considered incidental, related to aging, or procedural, were observed in both the untreated and intravaginally treated groups. The proportion of animals with malignant tumors, the total number of malignant tumors, as well as the types of malignant tumors in the three groups was similar. The cumulative incidence of microscopic lesions in various organs showed that malignant lymphoma was the major cause of death in aging female B 6C 3F 1 mice, the incidence of which was unaffected by intravaginal treatment. We conclude that long-term intravaginal administration of WHI-07 is not associated with systemic toxicity or increased carcinogenicity in mice. WHI-07 has clinical potential as an active ingredient of a safe vaginal/rectal microbicide.

Subchronic (13-week) toxicity studies of intravaginal administration of spermicidal vanadocene acetylacetonato monotriflate in mice

Bis-cyclopentadienyl complexes of vanadium(IV) or vanadocenes are rapid and potent inhibitors of human sperm motility with potential as a new class of contraceptive agents. In this study, groups of 10 B 6C 3F 1 and 20 CD-1 female mice were exposed intravaginally to a gel-microemulsion containing 0, 0.06, 0.12, or 0.25% of a representative vanadocene, vanadocene acetylacetonato monotriflate (VDACAC), five days per week for 13 consecutive weeks. The doses of VDACAC used were nearly 300- to 1250-fold higher than its in vitro spermicidal EC 50 value. After 13 weeks of intravaginal treatment, B 6C 3F 1 mice were evaluated for survival, body weight gain, absolute and relative organ weights, and systemic toxicity. Blood was analyzed for hematological and clinical chemistry profiles. Microscopic examination was performed on hematoxylin- and eosin-stained tissue sections from each study animal. Vanadium content in tissues was determined by atomic absorption spectroscopy. Gel-microemulsion (placebo) control and VDACAC dosed female CD-1 mice were mated with untreated males in order to evaluate if VDACAC has any adverse effects on the reproductive outcome. There were no treatment-related mortalities in either study. Mean body weight gain during the dosing period was not reduced by VDACAC treatment. Hemograms or clinical chemistry profiles did not reveal any toxicologically significant changes attributed to VDACAC treatment. No clinically significant dose-dependent changes in absolute and relative organ weights were noted in VDACAC dose groups. Extensive histopathological examination of tissues revealed no treatment-related abnormalities in any of the three VDACAC dose groups. Vanadium was not incorporated in mouse tissues at levels above 1 μg/g. Repeated intravaginal exposure of CD-1 mice to increasing concentrations of VDACAC for 13 weeks had no adverse effect on their subsequent reproductive capability (100% fertile), neonatal survival (>96%) or pup development. Collectively, these findings demonstrate that repetitive intravaginal administration of VDACAC to yield effective spermicidal concentrations (<0.1%) in the vagina was not associated with systemic toxicity and did not adversely affect the reproductive performance in mice. The spermicidal vanadocene-chelated complex, VDACAC, may be useful as a safe vaginal contraceptive.

Clustering of Hepatotoxins Based on Mechanism of Toxicity Using Gene Expression Profiles

Microarray technology, which allows one to quantitate the expression of thousands of genes simultaneously, has begun to have a major impact on many different areas of drug discovery and development. The question remains of whether microarray analysis and gene expression signature profiles can be applied to the field of toxicology. To date, there are very few published studies showing the use of microarrays in toxicology and important questions remain regarding the predictability and accuracy of applying gene expression profiles to toxicology. To begin to address these questions, we have treated rats with 15 different known hepatotoxins, including allyl alcohol, amiodarone, Aroclor 1254, arsenic, carbamazepine, carbon tetrachloride, diethylnitrosamine, dimethylformamide, diquat, etoposide, indomethacin, methapyrilene, methotrexate, monocrotaline, and 3-methylcholanthrene. These agents cause a variety of hepatocellular injuries including necrosis, DNA damage, cirrhosis, hypertrophy, and hepatic carcinoma. Gene expression analysis was done on RNA from the livers of treated rats and was compared against vehicle-treated controls. The gene expression results were clustered and compared to the histopathology findings and clinical chemistry values. Our results show strong correlation between the histopathology, clinical chemistry, and gene expression profiles induced by the agents. In addition, genes were identified whose regulation correlated strongly with effects on clinical chemistry parameters. Overall, the results suggest that microarray assays may prove to be a highly sensitive technique for safety screening of drug candidates and for the classification of environmental toxins.

Short-Term (13-week) toxicity study of 5-bromo-6-methoxy-5,6-dihydro-3'-azidothymidine-5'-(p-bromophenyl) methoxyalaninyl phosphate (WHI-07), a novel anti-HIV and contraceptive agent, in B6C3F1 mice.

The zidovudine derivative, WHI-07 (5-bromo-6-methoxy-5,6-dihydro-3'-azidothymidine-5'-(p-bromophenyl) methoxyalaninyl phosphate), is a dual-function spermicidal agent with potent anti-HIV activity. In this study, groups of 20 female B6C3F1 mice were exposed intravaginally to a gel microemulsion containing 0, 0.5, 1.0, or 2.0% WHI-07, 5 days per week for 13 consecutive weeks. On a molar basis, these concentrations of WHI-07 are 1400- to 5700-times higher than its spermicidal EC(50) and 1.4 x 10(6) to 5.7 x 10(6) times higher than its in vitro anti-HIV IC(50). After 13 weeks of intravaginal treatment, mice were evaluated for toxicity. The endpoints that were used for evaluation included survival, body weight, hematologic and clinical chemistry profiles, absolute and relative organ weights, and histopathology. No effects related to WHI-07 treatments were observed on survival, mean body weight, and mean body-weight gain. Repeated intravaginal exposure of mice to WHI-07 for 13 weeks had no toxicologically significant effect on organ weights, and did not cause any adverse changes in hematology parameters or blood chemistry profiles. Extensive histopathologic examination of tissues showed no lesions of pathologic significance. Thus, intravaginal application of WHI-07, for up to 13 weeks, does not cause systemic toxicity.

Intravaginal Toxicity Studies of a Gel-Microemulsion Formulation of Spermicidal Vanadocenes in Rabbits

Bis-cyclopentadienyl complexes of vanadium(IV) or vanadocenes are rapid and potent inhibitors of human sperm motility with potential as a new class of contraceptive agents. We investigated the toxicity potential of intravaginally administered gel-microemulsion formulation of two representative vanadocenes, vanadocene acetylacetonato monotriflate (VDACAC) and vanadocene dithiocarbamate (VDDTC), in the rabbit model. New Zealand White rabbits in subgroups of three were exposed intravaginally to a gel-microemulsion with and without 0.1 or 0.25% VDACAC and VDDTC for 10 consecutive days. The doses of vanadocenes used were nearly 500- to 1250-fold and 2000- to 5000-fold higher than their respective in vitro spermicidal EC50 values. Animals were euthanized on day 11 and vaginal tissues were evaluated for local toxicity by histopathology, cell proliferating activity by immunohistochemical detection of proliferating cell nuclear antigen (PCNA), and in situ apoptosis by the terminal deoxynucleotidyl transferase-mediated FITC-deoxyuridine triphosphate nick end-labeling (TUNEL) assay and confocal laser scanning microscopy (CLSM). Blood was analyzed for clinical chemistry profiles. Vanadium content in selected organs and body fluids was determined by atomic absorption spectroscopy. None of the rabbits given 0.1% VDACAC and VDDTC intravaginally developed epithelial ulceration, edema, leukocyte influx, or vascular congestion characteristic of inflammation. Only minimal to moderate irritation was observed at 0.25% VDACAC and VDDTC. A significant decrease in epithelial and stromal PCNA expression was observed in the 0.25% dose group. However, TUNEL assay and CLSM revealed no staining in the vaginal epithelium and only minimal nonspecific staining in the stroma. Repetitive intravaginal application of 0.1 or 0.25% VDACAC and VDDTC had no adverse effects on clinical chemistry profiles. Vanadium was not incorporated into rabbit tissues and body fluids at levels above 1 μg/g. Thus, intravaginal administration of VDACAC and VDDTC at concentrations nearly 500 and 2000 times higher than their respective in vitro spermicidal EC50 values did not induce marked vaginal irritation, mucosal toxicity, or systemic absorption of vanadium in the rabbit model. The lack of significant mucosal or systemic toxicity of intravaginal vanadocenes observed may have particular clinical utility as a new class of contraceptive agents.

The acute and subchronic toxicity of BRB-I-28, a novel class Ib antiarrhythmic agent, in CD-1 mice

The acute and subchronic toxic effects of BRB-I-28 (7-benzyl-3-thia-7-azabicyclo[3.3.1]nonane HCl), a novel class Ib antiarrhythmic agent, were investigated in male and female mice. The estimated oral LD 50 for BRB-I-28 was 128 mg/kg (male mice) and 131 mg/kg (female mice). In subchronic oral studies, four groups of mice (15/sex/group/dose) were fed daily with diets containing BRB-I-28 for 90 consecutive days. The equivalent daily doses were approximately 0, 16, 32, 76 (male) and 0, 18, 37, 89 mg/kg (female). All mice survived. Food consumption per day was decreased, but water consumption per day was increased (in a non-dose-dependent manner). However, both mean body weight and mean body weight gain were not significantly changed as were true for hematological and clinical chemistry profiles, except for serum Na + concentration (male) and serum K + concentration in male and female mice (high dose levels). Hepatocellular necrosis occurred in male and female mice (in a dose-dependent fashion). Renal cortical vacuoles and myocardial necrosis with low numbers of lymphocytic infiltrations were present in female mice (middle and high doses). Lesions in the liver, kidney and heart were mild with (very small) changes in serum biochemical values. These data suggest that BRB-I-28 has limited toxic potential, and coupled with low proarrhythmic and other desirable cardiovascular effects, makes BRB-I-28 worthy of further development.

Blood values in wild and captive Komodo dragons (Varanus komodoensis).

The Komodo dragon (Varanus komodoensis) is the largest living lizard and occupies a range smaller than that of any other large carnivore in the world. Samples from 33 free-ranging animals at five localities in Komodo National Park, Indonesia were evaluated to assess underlying health problems. To build a comparative database, samples from 44 Komodo dragons in both Indonesian and U.S. zoos were also analyzed. Tests performed included complete blood counts, clinical chemistry profiles, vitamin A, D(3), and E analyses, mineral levels, and screening for chlorinated pesticides or other toxins in wild specimens. Blood samples from wild dragons were positive for hemogregarines, whereas captive specimens were all negative. Total white blood cell counts were consistently higher in captive Komodo dragons than in wild specimens. Reference intervals were established for some chemistry analytes, and values obtained from different groups were compared. Vitamin A and E ranges were established. Vitamin D(3) levels were significantly different in Komodo dragons kept in captive, indoor exhibits versus those with daily ultraviolet-B exposure, whether captive or wild specimens. Corrective measures such as ultraviolet-permeable skylights, direct sunlight exposure, and self-ballasted mercury vapor ultraviolet lamps increased vitamin D(3) concentrations in four dragons to levels comparable with wild specimens. Toxicology results were negative except for background-level chlorinated pesticide residues. The results indicate no notable medical, nutritional, or toxic problems in the wild Komodo dragon population. Problems in captive specimens may relate to, and can be corrected by, husbandry measures such as regular ultraviolet-B exposure. Zoo Biol 19:495-509, 2000. Copyright 2000 Wiley-Liss, Inc.

Evaluation of subchronic (13-week) and reproductive toxicity potential of intravaginal gel-microemulsion formulation of a dual-function phenyl phosphate derivative of bromo-methoxy zidovudine (compound whi-07) in B(6)C(3)F(1) mice.

Heterosexual transmission of human immunodeficiency virus (HIV) accounts for 90% of all new infections worldwide and significantly contributes to new acquired immunodeficiency syndrome (AIDS) cases in the USA. In a systematic effort to develop a microbicidal contraceptive capable of preventing HIV transmission as well as providing fertility control, we previously identified novel phenyl phosphate derivatives of 3'-azido-3'-deoxythymidine (zidovudine) that exhibit potent anti-HIV and spermicidal activities. This study reports the preclinical studies of our lead compound WHI-07, 5-bromo-6-methoxy-5,6-dihydro-3'-azidothymidine-5'-(p-bromophenyl) methoxyalaninyl phosphate, for use as a dual-function topical microbicide. In vivo toxicity studies in non-human primates and rodents given WHI-07 (20 mg kg(-1)) intravenously and intraperitonealy, respectively, had no detectable adverse effects on hematological and clinical chemistry profiles. The 13-week subchronic and reproductive toxicity potential of an intravaginal gel-microemulsion formulation of WHI-07 was studied in mice to support its further development as a dual-function microbicide. Groups of ten female B(6)C(3)F(1) mice were exposed intravaginally to a gel-microemulsion formulation containing 0, 0.5, 1.0 or 2.0% WHI-07, 5 days a week, for 13 consecutive weeks. On a molar basis, these concentrations represent 1400-5700 times their in vitro spermicidal potency EC(50)) and 1.4 x 10(6)-5.7 x 10(6) times their in vitro anti-HIV activity(50)). After 13 weeks of intravaginal treatment, half of the treated mice were evaluated for toxicity and the other half were mated with untreated males to evaluate potential reproductive and developmental effects. The endpoints that were evaluated included survival, body weight gain, hematological and clinical chemistries, absolute and relative organ weights and histopathology. The WHI-07 applications did not cause weight loss, morbidity, mortality or specific tissue lesions detectable by histopathology. Repeated intravaginal exposure of mice to WHI-05 for 13 weeks had no adverse effects on subsequent reproductive performance (100% fertile), neonatal survival (>95%) or pup development. These findings collectively show that the experimental dual-function anti-HIV and contraceptive agent WHI-07 did not cause significant acute or subchronic toxicity.

Clinical outcome of a protocol to produce immunosuppression in rhesus monkeys (Macaca mulatta): application to infectious disease and gene therapy studies.

Induced immunosuppression is required for a number of studies using rhesus monkeys (Macaca mulatta). This report describes the clinical outcome and safety of a dose-finding experiment that determined doses of cyclophosphamide and prednisone that could be used to induce a state of immunosuppression in rhesus monkeys. After determining the optimum dose of immunosuppressive agents, the protocol was then used on animals participating in infectious disease and gene therapy studies. Splenectomy was performed in some animals to increase the severity of immunosuppression. The onset, duration, and severity of lymphopenia and leukopenia were consistent in all animals. In most animals, physical examination findings and clinical serum chemistry profiles demonstrated only transient abnormalities. With proper clinical monitoring, combination treatment with cyclophosphamide and prednisone is an effective and safe method for inducing immunosuppression in rhesus monkeys.

An evaluation of the toxicities of 2'-fluorouridine and 2'-fluorocytidine-HCl in F344 rats and woodchucks (Marmota monax).

The toxicities of 2'-fluorouridine (2'-FU) and 2'-fluorocytidine-HCl (2'-FC) were separately evaluated in 2 species, male Fischer 344 (F334) rats and woodchucks. Particular attention was focused on the ability of these nucleosides to induce toxicities similar to those induced by the antiviral drug fialuridine (FIAU). 2'-FU or 2'-FC was administered to F344 male rats by intravenous injection at doses of 5, 50, and 500 mg/kg/day for 90 consecutive days and to male and female woodchucks at doses of 0.75 and 7.5 mg/kg/day for 90 consecutive days. Clinical chemistry, hematology, and urinalysis (woodchuck only) profiles were assessed during and at the termination of the study. At necropsy, organs were weighed and tissues collected for routine histologic analysis. Cytochrome c oxidase activity, citrate synthase activity, and mitochondrial DNA content were measured, and micronucleus formation in the bone marrow (rats only) was evaluated. No adverse clinical effects were observed in either species. Rats treated with high doses of either 2'-FU or 2'-FC had body weights that were 90% of those of controls. 2'-FU and 2'-FC both induced a moderate decrease in the median lymphocyte count, and 2'-FC and 2'-FU induced a mild increase in mean corpuscular hemoglobin and mean corpuscular volume. Both compounds caused slight to moderate, reversible, histologic changes in the spleen and thymus. In the woodchuck, 2'-FC caused a slight increase in mean absolute lymphocytes, and 2'-FC and 2'-FU slightly increased hepatic periportal vacuolation and/or mononuclear cell infiltration. In summary, neither compound showed evidence of the toxicity induced by fialuridine in either species. Although compound effects were observed, none of these effects were considered to be adverse, and the no-observed adverse effect level was determined to be 500 mg/kg/day for both compounds in the male F344 rat and 7.5 mg/kg/day in the woodchuck.

Reference intervals and physiologic alterations in hematologic and biochemical values of free-ranging desert tortoises in the Mojave Desert.

Desert tortoise (Gopherus agassizii) populations have experienced precipitous declines resulting from the cumulative impact of habitat loss, and human and disease-related mortality. Evaluation of hematologic and biochemical responses of desert tortoises to physiologic and environmental factors can facilitate the assessment of stress and disease in tortoises and contribute to management decisions and population recovery. The goal of this study was to obtain and analyze clinical laboratory data from free-ranging desert tortoises at three sites in the Mojave Desert (California, USA) between October 1990 and October 1995, to establish reference intervals, and to develop guidelines for the interpretation of laboratory data under a variety of environmental and physiologic conditions. Body weight, carapace length, and venous blood samples for a complete blood count and clinical chemistry profile were obtained from 98 clinically healthy adult desert tortoises of both sexes at the Desert Tortoise Research Natural area (western Mojave), Goffs (eastern Mojave) and Ivanpah Valley (northeastern Mojave). Samples were obtained four times per year, in winter (February/March), spring (May/June), summer (July/August), and fall (October). Years of near-, above- and below-average rainfall were represented in the 5 yr period. Minimum, maximum and median values, and central 95 percentiles were used as reference intervals and measures of central tendency for tortoises at each site and/or season. Data were analyzed using repeated measures analysis of variance for significant (P < 0.01) variation on the basis of sex, site, season, and interactions between these variables. Significant sex differences were observed for packed cell volume, hemoglobin concentration, aspartate transaminase activity, and cholesterol, triglyceride, calcium, and phosphorus concentrations. Marked seasonal variation was observed in most parameters in conjunction with reproductive cycle, hibernation, or seasonal rainfall. Year-to-year differences and long-term alterations primarily reflected winter rainfall amounts. Site differences were minimal, and largely reflected geographic differences in precipitation patterns, such that results from these studies can be applied to other tortoise populations in environments with known rainfall and forage availability patterns.

D-Dimer Testing within the Routine Clinical Chemistry Profile

D-Dimer is the term for any of a variety of split products that are derived from fibrin fibrils by the action of the fibrinolytic enzyme plasmin. They occur in the circulation in a wide range of molecular weights and carry various numbers of cross-linked D-domains, called D-Dimers (1). Although no commonly accepted standard is available, D-Dimers can be measured by immunoassays that use specific antibodies against the cross-linked D-domain. The concentration of D-Dimers in the blood plasma is correlated to the activity state of the coagulation and fibrinolytic system (2). D-Dimer testing has been shown to be of value for diagnosis of clinically suspected thromboembolism (2)(3)(4)(5). In a recent meta-analysis (6) including data from 2000 patients with suspected deep venous thrombosis and/or pulmonary embolism, increased D-Dimers as measured by ELISA were found to provide a diagnostic sensitivity of 97% for both diseases. However, the practical diagnostic value of D-Dimer testing by ELISA is limited by the long turnaround time and the special equipment required for the assay. Qualitative or semiquantitative latex agglutination slide-assays are suited for bedside testing, but are not …

EFFECTS OF PYRUVATE SUPPLEMENTATION DURING TRAINING ON HEMATOLOGIC AND METABOLIC PROFILES

881 Pyruvate supplementation has been reported to promote fat mass loss in sedentary obese individuals possibly by altering carbohydrate and/or lipid metabolism. The purpose of this study was to examine the effects of pyruvate supplementation (10 g/d) during training on standard clinical hematological and metabolic profiles. 23 healthy females (33±2 yrs, 71.5±2 kg, 27.4±0.7 BMI, 34.9±1% body fat) were matched/paired and assigned to ingest two times per day for 30 days in a double-blind and randomized manner either a placebo (PL) containing 2.5 g calcium carbonate, 4 g maltodextrin, & 1 g dextrose or 5 g/d of calcium pyruvate (PYR). During this time, subjects participated in a supervised walking (30 min, 3 times per week at 70% of VO2max) and resistance-training program (2 sets of 8-12 repetitions, 8 machines, 3 times per week). Prior to and following supplementation, fasting blood samples were obtained. A comprehensive clinical chemistry profile was run on serum samples and whole blood samples were analyzed for cell blood counts. Data are presented as mean±SEM percent changes from pre-supplementation values for the PL and PYR groups, respectively. ANOVA for repeated measures revealed no significant (p<0.05) or clinically meaningful differences between groups in urea nitrogen, creatinine, the ratio of urea nitrogen to creatinine, uric acid, glucose, electrolyte status, total protein, albumin, globulin, carbon dioxide, alkaline phosphatase, hemoglobin, hematocrit, red blood cells, white blood cells, or types of lymphocytes. There was some evidence that muscle and liver enzymes following training were lower in the PYR group (CK 11±18, 10±11%, p=0.97; LDH 4±5, -9±4%, p=0.06; AST 16±7,-1±7%, p=0.13; ALT 23±17, -21±9%, p=0.03; GGT 10±12, -7±6%, p=0.06). There was also evidence that PYR supplementation may have negated some of the positive lipid modifying effects of training (CHL 3±4, -2±4%, p=0.38; HDL 14±6,-6±4%, p=0.01; CHL/HDL ratio -8±5, 4±4%, p=0.06; LDL 4±7, -7±5%, p=0.25; VLDL -9±9, 19±17%, p=0.17; TG-13±7, 18±18%, p=0.13). Results indicate that although most hematological markers were not affected, additional study is necessary to evaluate the effects of PYR supplementation on lipid profiles and to determine whether the changes observed may limit the usefulness of this nutrient as a weight loss supplement.

Oral Antiplatelet Efficacy of the Platelet GPIIb/IIIa Antagonist, DMP754 in Non-Human Primates

Binding kinetic studies with XV459, the active form of DMP754, demonstrated comparable binding kinetics (Kd and Koff) with platelets obtained from either human or baboons which were different from that with platelets obtained from dogs. Therefore, the present study was undertaken to evaluate the antiplatelet efficacy of DMP754 following oral administration in baboons. The dose levels evaluated were 0.1 and 1.0 mg/kg, IV and 0.1, 0.3, 1.0, and 3.0 mg/kg, oral of DMP754. Oral doses of DMP754 resulted in dose- and time-related inhibition of platelet aggregation along with a modest effect on bleeding time prolongation. DMP754 at similar oral doses had 24 hours of antiplatelet effects in baboon as compared to 8–12 hours duration of antiplatelet efficacy in dogs. At maximal antiplatelet doses DMP754 demonstrated no significant effects on platelet count, clinical chemistry or hemodynamic profiles in baboons. These data suggest that DMP754 is a potent orally active antiplatelet agent with extended duration after once a day oral administration in non-human primate.

Reproductive effects of fenoxycarb on sheep.

Fenoxycarb (ethyl [2-(4-phenoxyphenoxy)-ethyl] carbamate) is an insect growth regulator used for long-term fire ant control. Because of its effects on insect reproduction and its potential use on pasturage consumed by food animals, a reproductive study was conducted using Rambouillet sheep. The sheep were dosed daily with a placebo or with fenoxycarb at 0.69 or 1.38 mg/kg/day, representing ten (10x) and 20 times (20x) the maximum amounts of fenoxycarb in forage or hay treated at recommended levels for fire ant control. Parameters that were measured included rates of weight gain of adults, serum clinical chemistry profiles of adults, spermatozoal morphology and motility, estrus cycling, pregnancy rates, maintenance of pregnancies to term, numbers of live births, and rates of weight gain of lambs to 28 days. There were no statistically significant (P < or = 0.05) differences between the exposed and control groups of sheep in any of these facets of the study. No clinical signs associated with exposure to fenoxycarb were observed in any animal at any time, and no exposure-related pattern of pathologic lesions or reproductive organ histology was observed. Means of hepatic fenoxycarb residues in the rams followed a statistically significant (P < or = 0.05) dose-related pattern. No fenoxycarb was detected (detection limit of 5 ppb) in any neonatal liver, despite the presence of hepatic fenoxycarb residues in the treated ewes, indicating that transplacental transport of fenoxycarb was minimal. No fenoxycarb was detected in any lamb liver at 28 days, although both the colostrum and the milk of exposed ewes were found to contain fenoxycarb at levels proportional to the treatments. Based on the lack of significant findings in this study, it is unlikely that use of fenoxycarb, according to label instructions (currently applicable to homeowner and registered agricultural usage) for fire ant control in pasturage or hay fields, will affect ruminant reproduction.

Effects of graded levels of bentonite on serum clinical profiles, metabolic hormones, and serum swainsonine concentrations in lambs fed locoweed (Oxytropis sericea)1

To determine which clay minerals have the potential to bind swainsonine, an in vitro screening procedure was conducted. Twenty compounds were screened in one replicated in vitro trial. A commercially available bentonite bound approximately 10% swainsonine and was chosen for use in a subsequent lamb feeding trial. Twenty fine-wool lambs (30.5 +/- .7 kg) were assigned to one of five treatments (four lambs/treatment). Treatments included 1) positive control, 100% sorghum sudangrass hay, 2) 85% sorghum sudangrass:15% locoweed (Oxytropis sericea, 430 ppm [DM basis] of swainsonine) +0 g of bentonite, 3) Treatment 2 + 14 g of bentonite, 4) Treatment 2 + 28 g of bentonite, and 5) Treatment 2 + 42 g of bentonite. Lambs were fed the experimental diets for 35 d and were then fed the positive control diet for an additional 21 d. Lambs were weighted and blood was collected via jugular venipuncture weekly from d 0 through 56. On d 35, additional blood samples were collected 1, 2, 4, and 8 h after feeding. Weekly blood samples were analyzed for serum clinical chemistry profiles, and additional blood samples collected on d 35 were analyzed for serum metabolic hormones and serum swainsonine concentrations. Within 1 wk, serum alkaline phosphatase and glutamic oxaloacetic transaminase activities increased markedly (P < .05) in lambs fed locoweed. Serum insulin, growth hormone, and prolactin concentrations were not affected by feeding locoweed, but serum triiodothyronine and thyroxine concentrations were decreased by approximately 50% (P < .05) in lambs fed locoweed.(ABSTRACT TRUNCATED AT 250 WORDS)

Effects of lasalocid and monensin plus tylosin on serum metabolic hormones and clinical chemistry profiles of beef steers fed a 90% concentrate diet.

Fourteen crossbred beef steers (average initial BW 313 +/- 13.1 kg) fed a 90% concentrate diet (as-fed basis) were used to evaluate effects of ionophores on serum metabolic hormones and clinical chemistry profiles. Treatments were no ionophore (C; four steers), lasalocid (L; 33 mg/kg of diet; five steers), and monensin (33 mg/kg of diet) plus tylosin (11 mg/kg of diet; MT; five steers). All steers were adapted to the 90% concentrate diet, after which treatments were applied and blood was sampled via jugular catheters on d 7, 35, 63, 91, and 119 of the trial at 0, 1, 2, 3, 4, 6, 8, and 10 h after feeding. Averaged over sampling days, serum glucose concentrations did not differ (P > .10) among treatments at any sampling time (treatment x sampling time, P < .05). Average across sampling days and times, serum growth hormone, insulin, cholesterol, and triglyceride concentrations did not differ (P > .10) among treatments. No major effects of ionophores were noted for serum enzymes or protein fractions. Averaged over sampling times within day (treatment x day interaction, P < .10), both L and MT increased (P < .05) serum Ca and Na concentrations on d 91 and 119 compared with C. Serum inorganic P was increased (P < .10) for L vs MT and for ionophore treatments compared with C (P < .10) on several occasions (treatment x day x sampling time interaction, P < .05). Results suggest that ionophores do not cause dramatic changes in serum metabolic hormones or clinical chemistry profiles; however, monensin and lasalocid altered serum minerals in beef steers fed a high-concentrate diet.