METASTATIC MINERALIZATION IN A ZOOLOGIC COLLECTION OF SPOT-TAILED EARLESS LIZARDS (HOLBROOKIASPP.).

Vitamin and mineral supplements are commonly used in diets for zoologic and companion animals. Because specific nutrient requirements are often unknown, informed decisions are based on literature for related species. Over 18 mon beginning in November 2017, an entire population of spot-tailed earless lizards (Holbrookia lacerata and Holbrookia subcaudalis) died (N = 33). All but two lizards were submitted for histopathology (94%). All examined cases had mineralization in at least one tissue; 71% (22 of 31) had multisystemic mineral deposits consistent with metastatic mineralization. No underlying causes were detected histologically. The supplement used for dusting the food items fed five to six times per week was inadvertently switched for 2 to 4 mon, and the incorrect supplement was found to contain fourfold the intended vitamin D3 concentration. Thus, hypervitaminosis D was considered the most likely cause. Interestingly, eastern collared lizards (Crotaphytus collaris), also fed prey supplemented five to six times a week, and over 50 other insectivorous reptile and amphibian species possibly receiving the supplement one to seven times a week did not appear affected. During this time, only two other cases of metastatic mineralization were diagnosed in other herpetofauna at this institution. Prior to receiving the incorrect supplement, there were no cases of metastatic mineralization detected in the earless lizard population. These cases highlight species-specific sensitivities, and the deleterious effects of excessive or inappropriate supplementation. It is important to confirm product identification on arrival, regularly conduct chemical analysis of supplements, and educate keepers and owners about adverse effects of inappropriate supplementation.

In Vitro Production of Radiolabeled Red Clover (Trifolium pratense) Isoflavones.

Red clover isoflavones are increasingly used in dietary supplements for their purported estrogenic effects. However, little is known about their metabolism in animals due to a lack of commercially available isotopically-labeled tracers. The goal of this research was to establish red clover cell culturing methodology for (14)C-biolabeling of isoflavones. When root, leaf, and petiole-derived suspension cultures were grown in darkness or light, dark-grown, petiole-derived solution cultures produced the highest concentrations of the two major red clover isoflavones, formononetin (0.67 mg/g FM inoculum) and biochanin A (0.13 mg/g FM inoculum). Varying levels and timing of copper chloride elicitor did not significantly affect isoflavone accumulation. Approximately 38% of the (14)C-sucrose dose accumulated in the cells. Eighteen percent of the initial labeled dose was detected in the isoflavone-rich methanolic extract and of that, 22% accumulated in isoflavones.

In vivo metabolic tracking of 14C-radiolabelled isoflavones in kudzu (Pueraria lobata) and red clover (Trifolium pratense) extracts.

Absorption, distribution and elimination of 14C-labelled isoflavone-containing extracts from kudzu (Pueraria lobata) root culture and red clover (Trifolium pratense) cell culture were investigated in an in vivo rat model. The predominant isoflavones in the kudzu extract were the glycosides puerarin, daidzin and malonyl daidzin, while in the red clover extract, the major isoflavones were formononetin and its derivatives, genistein and biochanin A, with radioactivities of 3.770 and 7.256 MBq/g, respectively. Male Sprague-Dawley rats, implanted with a jugular catheter and a subcutaneous ultrafiltrate probe, were orally administered with 14C-labelled isoflavone extracts from either kudzu or clover cell cultures. Serum, interstitial fluid (ISF), urine and faeces were collected using a Culex Automated Blood Collection System for 24 h. Analysis of bone tissues revealed that radiolabel accumulated in the femur, tibia and vertebrae at 0.04, 0.03 and 0.01 % of the administered dose, respectively, in both kudzu and red clover treatments. The liver accumulated the greatest concentration of radiolabel among the tissues tested, at 1.99 and 1.54 % of the administered kudzu and red clover extracts, respectively. Serum and ISF analysis showed that both extracts were rapidly absorbed, distributed in various tissues, and largely eliminated in the urine and faeces. Urine and faeces contained 8.53 and 9.06 % of the kudzu dose, respectively, and 3.60 and 5.64 % of the red clover dose, respectively. Serum pharmacokinetics suggest that extracts from kudzu may undergo enterohepatic circulation.

Prevalence of metabolic syndrome and associated risk factors in Illinois.

PURPOSE: To examine the prevalence of metabolic syndrome (MS) in Illinois and associated risk factors to document emerging problems as a basis for preventive and/or therapeutic interventions. DESIGN: The 2005 Illinois Behavioral Risk Factor Surveillance System data were analyzed for prevalence rates and relative risk (RR) of risk factors for MS. SETTING: Illinois. SUBJECTS: Representative sample of 5077 noninstitutionalized Illinoisans aged > or = 18. MEASURES: Presence of diabetes, hypertension, hypercholesterolemia, angina, and obesity; adequate fruit/vegetable intake; adequate daily physical activity; routine consumption of calcium-rich foods; smoking; and demographic variables. ANALYSIS: Prevalence, chi-square, RR, and analyses of variance. RESULTS: MS prevalence was 16.2%. The RR for MS was 8 for those having diabetes; 20 for those with hypertension; 14 for those with hypercholesterolemia; 6 for those with angina; and 7 for those with obesity. Fruit/vegetable intake did not significantly influence the RR (RR = 1.01), but RR was greater when calcium-rich foods were not routinely consumed (RR = 1.61) and with inadequate physical activity (RR = 1.85). MS prevalence was highest in those over 65 years; with incomes < $15,000; and among Blacks. Differences among those with one through five indicators of MS were not significant for fruit/vegetable intake, but were for physical activity, age group, income, and education level (p < .001). CONCLUSION: These findings reveal that MS prevention should be emphasized for lower-income, older Blacks. Increasing physical activity and calcium-rich foods are areas for community-based education.

Isolation of radiolabeled isoflavones from kudzu (Pueraria lobata) root cultures.

Isoflavones have potential for preventing and treating several chronic health conditions, such as osteoporosis, cardiovascular disease, and cancer. In this study, radiolabeled isoflavones were recovered from kudzu (Pueraria lobata) root cultures after incubation with uniformly labeled (14)C-sucrose in the culture medium for 21 days. Approximately 19% of administered label was recovered in the isoflavone-rich dried extracts of kudzu root cultures (90.2 microCi/g or 3.3 MBq/g extract). HPLC-PDA analysis revealed the predominant isoflavones isolated from kudzu root cultures to be puerarin, daidzin, and malonyl-daidzin. The average concentration of the major isoflavone puerarin in kudzu root cultures was 33.6 mg/g extract, with a specific activity of 63.5 microCi/g (2.3 MBq/g). The isolated isoflavones were sufficiently (14)C-labeled to permit utilization for subsequent in vivo metabolic tracking studies.

Identification of isoflavone glycosides in Pueraria lobata cultures by tandem mass spectrometry.

Isoflavones in the methanolic extracts of kudzu (Pueraria lobata) callus, suspension and root cultures were compared in order to develop an experimental system in which puerarin (daidzein 8-C-glucoside) and other isoflavones could be synthesised in vitro. Quantitative variation of puerarin and other known isoflavones was estimated in kudzu culture extracts using HPLC-UV. The highest and lowest amounts of puerarin (14.56 and 0.33 mg/g) were present in in vitro root cultures and leaf tissue-derived callus cultures, respectively. A total of 48 isoflavone metabolites were detected in extracts of kudzu root cultures by HPLC-MS/MS, and the structures of 33 of them were tentatively assigned. Amongst these, 12 isoflavone C-glycosides were identified. Hydroxyderivatives of puerarin in several isomeric forms were detected, some of which have not been previously reported in kudzu root. The molecular weights, interpretation of characteristic fragment ions obtained from HPLC-MS/MS and comparison with reported data allowed the putative identification of the isoflavone metabolites.