Harvesting Sugar From Nonflowering Plants: Implications of a Marked Sugar Bait on Honey Bee (Hymenoptera: Apidae) Whole Hive Health.

Honey bees (Apis mellifera) are effective foragers for floral and extra-floral sources of sugars and as a result may easily be exposed to toxicants in the environment, such as pesticides. Toxic sugar baits (TSB) or insecticide-laced baits are designed for insect vector management but may be a danger to foraging honey bees and their hives. During a mosquito movement study at a zoological institution, nonflowering foliage surrounding the zoo was marked with sugar solution dyed with over the counter food-coloring. Mosquitoes and other insects foraged on the marked sugar and were collected within the zoo. Additionally, four of six honey bee hives within the zoo had workers that also foraged the dyed sugar and consequently colored approximately 57 kg of honey within the hives. This observation identifies a possibility route of toxicant accumulation within the hives from TSB and possible exposure may have whole hive health implications not previously described on an individual bee level.

Development of a novel clinical biomarker assay to detect and quantify aggrecanase-generated aggrecan fragments in human synovial fluid, serum and urine.

OBJECTIVE: Proteolytic degradation of aggrecan in articular cartilage is a hallmark feature of osteoarthritis (OA). The present study was aimed at developing a sensitive enzyme linked immunosorbent assay (ELISA) for the detection of aggrecanase-cleaved fragments of aggrecan in human serum and urine to facilitate the clinical development of aggrecanase inhibitors for OA. METHODS: The BC3 monoclonal antibody that detects the ARGS neoepitope sequence in aggrecanase-cleaved aggrecan was engineered and optimized using complementarity determining region (CDR)-saturation mutagenesis to improve its binding affinity to the neoepitope. A sandwich ELISA (BC3-C2 ELISA) was developed using the optimized alpha-ARGS antibody (BC3-C2) as capture antibody and a commercially available antibody directed against the hyaluronic-acid binding region (HABR) of aggrecan as detection antibody. Aggrecanase-cleaved fragments of aggrecan present in in vitro digests, human cartilage explant culture supernatants and in human synovial fluid, serum and urine were detected and quantified using this ELISA. RESULTS: The optimized antibody had a 4-log improvement in affinity for the ARGS containing peptide compared to the parental BC3 antibody, while maintaining the ability to not cross-react with a spanning peptide. The BC3-C2 ELISA demonstrated the ability to detect aggrecanase-cleaved aggrecan fragments in the native state, without the need for deglycosylation. This ELISA was able to measure aggrecanase-generated ARGS containing aggrecan fragments in human articular cartilage (HAC) explant cultures in the basal state (without cytokine stimulation). Treatment with an aggrecanase inhibitor resulted in a dose-dependent inhibition of ARGS neoepitope released into the culture supernatant. The ELISA assay also enabled the detection of ARGS containing fragments in human synovial fluid, serum and urine, suggesting its potential utility as a biomarker of aggrecanase activity. CONCLUSIONS: We have developed a novel ELISA using an optimized ARGS antibody and have demonstrated for the first time, an ELISA-based measurement of aggrecan degradation products in human serum and urine. This assay has the potential to serve as a mechanistic drug activity biomarker in the clinic and is expected to significantly impact/accelerate the clinical development of aggrecanase inhibitors and other disease modifying drugs for OA.

Translocation as a species conservation tool: status and strategy.

Surveys of recent (1973 to 1986) intentional releases of native birds and mammals to the wild in Australia, Canada, Hawaii, New Zealand, and the United States were conducted to document current activities, identify factors associated with success, and suggest guidelines for enhancing future work. Nearly 700 translocations were conducted each year. Native game species constituted 90 percent of translocations and were more successful (86 percent) than were translocations of threatened, endangered, or sensitive species (46 percent). Knowledge of habitat quality, location of release area within the species range, number of animals released, program length, and reproductive traits allowed correct classification of 81 percent of observed translocations as successful or not.

Radioactivity of the lunar surface.

Diffusion of radon and thoron from the Ilunar surface provides a mechanism for production of a radioactive surface layer on the moon. If the radon and thoron flux from the lunar surface is equal to that measured at the earth's surface, the equilibrium activity of this surface layet is estimated as approximately 1 microcurie per square meter, due to radon and its decay products. This activity consists of alpha particles and gamnmna rays at well-defined energies and of beta rays.

Single and multiple-dose pharmacokinetics of tepoxalin and its active metabolite after oral administration to rabbits (Oryctolagus cuniculus).

The anti-inflammatory agent, tepoxalin, was administered to eight healthy 6-month-old female New Zealand white rabbits once daily at an oral dose of 10 mg/kg. Blood samples were obtained immediately before and at 0.25, 0.5, 1, 2, 3, 4, 6, 8, 12, and 24 h postadministration on days 1 and 10. Tepoxalin and its active metabolite, RWJ 20142, concentrations were determined in plasma by use of high-performance liquid chromatography with mass spectrometry. C(max) of the parent compound was reached between 3 and 8 h of drug administration, with a harmonic mean t(1/2) of 3.6 h. Peak tepoxalin plasma concentrations were 207 +/- 49 ng/mL. After oral administration, the metabolite RWJ 20142 achieved C(max) in plasma 2-8 h after administration, with a t(1/2) of 1.9-4.8 h (harmonic mean 2.8 h). Peak plasma concentrations of RWJ 20142 on day 1 were 2551 +/- 1034 ng/mL.

Suspected Hepadnavirus Association with a Hepatocellular Carcinoma in a Black-Tailed Prairie Dog (Cynomys ludovicianus).

Hepatocellular carcinomas are the most commonly reported neoplasm of black-tailed prairie dogs (Cynomys ludovicianus). In several other closely related Sciuridae species, infection with species-specific hepadnaviruses is associated with the development of these tumours, but such a hepadnavirus has not yet been identified in any prairie dog species, although its presence has been hypothesized previously. An adult prairie dog was humanely destroyed due to progressive illness and the identification of a cranial abdominal mass that was determined on histopathology to be a hepatocellular carcinoma. Deep sequencing of the tumour tissue identified the presence of a hepadnavirus, similar in its genetic structure to woodchuck hepatitis virus. Electron microscopy showed the presence of viral particles similar in structure to other hepadnaviral particles. This report suggests that a hepadnavirus may be associated with the development of hepatocellular carcinomas in the prairie dog.

Mechanisms of action of 6-thioguanine, 6-mercaptopurine, and 8-azaguanine.

The effects of 6-thioguanine on purine biosynthesis and cell viability have been examined in H.Ep. 2 cells grown in culture. Toxicity is not reversed by aminoimidazolecarboxamide, suggesting that inhibition of purine biosynthesis de novo is not the sole mechanism of toxicity. Also, 6-(methylmercapto)purine ribonucleoside, a potent inhibitor of purine biosynthesis de novo, produces more marked reductions in cellular pools of purines than does 6-thioguanine without killing cells. There is no apparent inhibition by 6-thioguanosine 5'-monophosphate of other enzymes leading to the synthesis of guanosine 5'-triphosphate as determined in whole cells by measurements of radioactive hypoxanthine or guanine incorporation. Inhibition of DNA synthesis by 1 mM thymidine protects cells from 6-mercaptopurine or 6-thioguanine but fails to protect cells from 8-azaguanine toxicity. On the other hand, inhibition of RNA synthesis by 6-azauridine plus deoxycytidine protects cells against 8-azaguanine but does not protect against 6-thioguanine or 6-mercaptopurine toxicity. In agreement with the in vitro data, arabinosylcytosine (a potent inhibitor of DNA synthesis) fails to protect mice against 8-azaguanine but has previously been shown to protect mice from 6-mercaptopurine or 6-thioguanine toxicity. The results support the hypotheses of others that incorporation into DNA (as 6-thioguanine nucleotide) is a mechanism of toxicity for these thiopurines, whereas 8-azaguanine is toxic due to its incorporation into RNA.

Influence of obesity, impaired glucose tolerance, and NIDDM on LDL structure and composition. Possible link between hyperinsulinemia and atherosclerosis.

The possible causes of the enhanced risk for coronary heart disease (CHD) were examined in morbidly obese women with normoglycemia, impaired glucose tolerance (IGT), and non-insulin-dependent diabetes mellitus (NIDDM) before and after gastric bypass surgery. Compared with age-matched lean women, plasma lipid and apolipoprotein concentrations of the obese women before surgery favored atherogenesis. The risk for CHD may further be exacerbated in the IGT and NIDDM groups by the prevalence of smaller and denser low-density-lipoprotein (LDL) particles. LDL size correlated negatively with plasma insulin levels independent of triglyceride levels, age, or body mass index (BMI). After surgery, BMI, plasma insulin, and triglyceride levels decreased, but LDL size increased, and LDL density decreased. Neither cholesterol nor LDL cholesterol levels were affected after surgery, but high-density-lipoprotein cholesterol was increased in all patients after surgery. Although the mechanisms underlying the changes in the properties of LDL could not be determined from this study, these changes appear to be of benefit in reducing CHD risk in these patients.

Influence of morbid obesity and non-insulin-dependent diabetes mellitus on high-density lipoprotein composition and subpopulation distribution.

The chemical composition, subpopulation distribution and peak hydrated density of high-density lipoprotein (HDL) were examined in 18 morbidly obese women with either normal glucose tolerance or with non-insulin-dependent diabetes mellitus (NIDDM), and in 15 age-matched lean control subjects. Similar measurements were made in the morbidly obese subjects after gastric bypass surgery for weight loss. In the diabetic group, HDL was relatively protein-enriched and cholesterol- and cholesterol ester-poor compared with the lean controls. The same trend was seen in the nondiabetic group, although the cholesterol ester difference was not significant. In both cases, both electrophoretic and density gradient analysis showed that plasma HDL contained more of the HDL3 and less of the HDL2 subfractions than that for the lean controls. Following surgery in the diabetic group, the protein percentage of HDL decreased, and that of cholesterol ester increased; for the nondiabetic group, the protein decreased and phospholipid increased. In the diabetic group, a shift of the electrophoretic HDL subpopulation distribution toward more HDL2 accompanied these changes. Average hydrated peak density of HDL shifted from the HDL3 to the HDL2 range for the diabetic group following surgery; for the nondiabetic group the peak density also decreased but still remained within the HDL3 range. In all cases, the differences seen were more pronounced among the diabetic patients. These changes in the properties of HDL after gastric bypass surgery may favorably influence the risk for coronary heart disease usually associated with diabetes.

Alterations in low-density lipoproteins in subjects with abdominal adiposity.

Abdominal adiposity, as indexed by the waist to hip girth ratio (WHR), has been associated with increased risk and incidence of coronary heart disease (CHD). The purpose of this study was to determine if this enhanced risk is related to alterations in the structure of low density lipoproteins (LDL). LDL were isolated from nine nonobese men with an average WHR of 1.046 and nine nonobese men with a WHR of 0.94, who were matched on age (45.6 +/- 2.7 v 47.7 +/- 2.3 mean +/- SEM) percent body fat (26.5 +/- 0.5 v 26.1 +/- 0.9), and body mass index (27.3 +/- 0.6 v 26.3 +/- 0.6). The average molecular weight of LDL from the subjects with a high WHR was lower than that of subjects with low WHR (2.70 v 3.02 x 10(6) d), the average hydrated density higher (1.050 v 1.040 g/mL), and the mobility (Rf) on 2% to 16% polyacrylamide gradient gel electrophoresis higher. Subfractionation by equilibrium density ultracentrifugation showed that the LDL of subjects with a high WHR was predominantly in the heavy density range (1.038 to 1.048 g/mL) compared with the LDL of subjects with low WHR, which was in the lighter density range (1.030 to 1.040 g/mL). Chemical analysis of the subfractions showed that the peak density fractions of LDL of subjects with a high WHR had a lower cholesterol to protein ratio than the peak density fractions of LDL of subjects with low WHR. Electron microscopy of these peak density fractions showed that LDL of subjects with high WHR was smaller than that of subjects with low WHR. These characteristics of LDL of subjects with abdominal adiposity closely resemble the properties of LDL of patients with documented CHD. It is concluded that the increased risk of CHD associated with abdominal adiposity may be due in part to the alterations in LDL characteristics, and that these alterations in LDL are independent of the degree of obesity.

Lipogenic potential of liver from morbidly obese patients with and without non-insulin-dependent diabetes.

Intra-abdominal liver biopsies were obtained during surgery from fasted obese patients with non-insulin-dependent diabetes mellitus (NIDDM), obese normoglycemic controls, and lean controls. Lipid synthesis was studied in freshly isolated hepatocytes and liver homogenates from the three groups of subjects. Incorporation of 3H2O into the lipids of hepatocytes was determined in the absence and presence of insulin (0.1 mumol/L). The activities of five enzymes involved in fatty acid synthesis, and the incorporation of 14C-glycerol-3-phosphate into lipids were determined in liver homogenates. Basal lipid synthesis by hepatocytes was not different in the three groups of patients. Insulin stimulated lipogenesis by 8% +/- 30% in the lean controls, 33% +/- 8% in the obese controls and 17% +/- 6% in the NIDDM patients. No significant differences in the activities of the five enzymes that are involved in de novo fatty acid synthesis among the three groups of patients were observed. Similarly, incorporation of 14C-glycerol-3-phosphate by liver homogenates, in the presence of saturating or submaximal concentrations of fatty acids, did not differ among the three groups. These results show that under the experimental conditions of this study, including the fasted state of the patients, the basal capacity of liver of NIDDM patients to synthesize fatty acids or glycerides is the same as that of liver from obese and lean controls. Thus, it is likely that an increase in fatty acid flux into a liver with normal lipogenic potential may contribute to the increased synthesis of triglycerides by the liver of these patients in vivo.(ABSTRACT TRUNCATED AT 250 WORDS)

The effects of exercise on lipogenic enzyme activity and glyceride synthesis by liver homogenates of diabetic rats.

The purpose of this study was to determine if exercise ameliorates the elevated levels of triglycerides in diabetic rats and also to determine if the capacity of liver to synthesize glycerides correlates with changes in plasma triglyceride levels. Forty female rats were divided into four groups: sedentary control, sedentary diabetic, exercised control, and exercised diabetic. Diabetes was induced by intravenous injection of alloxan (40 mg/kg), and control rats were sham-dosed with physiologic saline. All rats remained sedentary in their cages for the first week after the injections. The exercised groups were exercised for seven consecutive days for 2 h/d at 20 m/min (0 grade). All rats were killed 24 hours after the last exercise bout. Blood glucose levels were significantly higher in the diabetic group than the nondiabetic counterparts, but exercise did not affect glucose levels in either the diabetic or nondiabetic groups. Exercise, however, significantly lowered plasma triglyceride and free fatty acid levels in both diabetic and nondiabetic rats. The activities of the five enzymes involved in fatty acid synthesis were all depressed in the diabetic rats compared to controls, and exercise had no effect on the activities of the enzymes. The capacity of liver to synthesize total lipids, diglycerides, or triglycerides was not different in the diabetic rats from that of nondiabetic rats, and exercise did not change that. Only phospholipid synthesis from glycerol-3-phosphate was affected by diabetes. It is concluded that exercise ameliorates the elevations in plasma triglyceride levels that result from diabetes.(ABSTRACT TRUNCATED AT 250 WORDS)

Influence of age and exercise training on lipid metabolism in Fischer-344 rats.

The influence of training on fatty acid and glyceride synthesis by liver and adipose tissue homogenates of young and old Fischer-344 rats was examined. Four groups of rats (10 animals/group) were studied: young untrained, young trained, old untrained, and old trained. Training of each group was for 10 wk at 75% maximal O2 uptake. Young rats were killed at 6 mo of age and old rats were killed at 27 mo of age. Fatty acid synthesis was assessed by measuring the activities of acetyl-CoA carboxylase, fatty acid synthase, ATP citrate-lyase, "malic" enzyme, and glucose-6-phosphate dehydrogenase. Glyceride synthesis was evaluated by determining the rate of incorporation of [14C]glycerol 3-phosphate into lipids. In addition, lipoprotein lipase activity was measured in acetone-ether powders of adipose tissue from the four groups of rats. In liver, training had no effect on fatty acid or glyceride synthesis in either group. However, aging caused a significant decrease in the activities of four of the lipogenic enzymes but had no effect on glyceride synthesis. Training caused an increase in fatty acid synthase and glyceride synthesis in adipose tissue, and aging decreased lipoprotein lipase activity. It was concluded that training enhances the synthetic capacity of lipids by adipose tissue but that aging had a more profound effect in that the activities of the enzymes involved in these processes were lower in the old rats. Furthermore, the decreased activity of lipoprotein lipase in the older rats may explain the higher plasma triglyceride levels that were observed in these animals.

Simultaneous extraction and quantitation of fentanyl and norfentanyl from primate plasma with LC/MS detection.

The quantitation of both fentanyl and its desalkyl metabolite, norfentanyl, in plasma using LC/MS has not been previously described. The detection and quantitation of fentanyl and norfentanyl was achieved using LC/MS detection. The liquid-liquid extraction used toluene as the organic phase. Chromatography was carried out using a Zirchrom-PBD (50 mm x 2.1 mm, 3 microm) column with a mobile phase of acetonitrile-ammonium acetate (10 mM), citrate (0.1 mM, pH 4.4) (45:55, v/v) with a flow rate of 0.3 ml/min. Mass spectroscopy detection was performed using ESI in the positive mode. The LOQ for fentanyl was 25 pg/ml and norfentanyl was 50 pg/ml. For the concentrations of 75, 250, and 750 pg/ml, respectively, fentanyl had inter-day precisions of 6.6, 7.2, and 6.6% with accuracies of 4.0, 5.1, and 5.1% and intra-day precisions of 1.6, 1.9, and 1.9% with accuracies of 11.6, 9.4, and 8.4%, and norfentanyl had inter-day precisions of 7.4, 0.3, and 0.7% with accuracies of 9.1, 8.8, and 12.3% and intra-day precisions of 5.3, 1.4, and 0.1% with accuracies of 10.9, 8.9, and 12.8%. The recoveries of fentanyl were 85, 92, and 75% and of norfentanyl were 40, 49, and 46% at the 75, 250, and 750 pg/ml concentrations, respectively.

Pharmacokinetics of gentamicin in blood plasma of quail, pheasants, and cranes.

Rate of appearance, peak concentration, and the biological half-life of gentamicin in the plasma of quail (Coturnix coturnix), pheasants (Phasianus colchicus), and cranes (Grus canadensis tabida) were studied. Gentamicin was given IM in doses of 5, 10, and 20 mg/kg of body weight. Peak plasma concentrations occurred earliest in the quail, latest in the cranes. The peak concentrations varied directly with the administered doses in all species. The biological half-life of gentamicin was 42 +/- 12 minutes in the quail, 75 +/- 15 minutes in the pheasants, and 165 +/- 37 minutes in the cranes. On the basis of the present data, dosage regimens for gentaminic of 5 mg/kg every 8 hours in pheasants and cranes, and 10 mg/kg every 6 hours in quail, would be expected to give constant plasma concentrations greater than 4.0 micrograms/ml.

Pharmacokinetics and tissue concentrations of tylosin in selected avian species.

Tissue and plasma concentrations and the biological half-life of tylosin in avian species of a variety of body sizes and metabolic rates were studied. The species chosen were eastern bobwhite quail (Colinus virginianus virginianus), pigeons (Columba livia), greater sandhill cranes (Grus canadensis tabida), and emus (Dromaius novaehollandiae). In the 1st phase of this study, tylosin was administered IM to quail, pigeons, and emus at a dosage rate of 25 mg/kg of body weight and to cranes at a dosage rate of 15 mg/kg. The average peak plasma concentrations of tylosin in quail, pigeons, cranes, and emus were 4.31, 5.63, 3.62, and 3.26 microgram/ml, respectively. These peak concentrations occurred at 0.5 to 1.5 hours after administration. The biological half-life of tylosin averaged 1.2 hours in quail, pigeons, and cranes, and was 4.7 hours in emus. In the 2nd phase of this study, tylosin concentrations in the tissues of quail, pigeons, and cranes were markedly higher than were plasma concentrations at corresponding sampling times. Six hours after antibiotic administration, tissue concentrations of tylosin in all species remained within the minimum inhibitory concentration for most pathogenic organisms. Dosage regimens of 25 mg of tylosin/kg 4 times daily for quail and pigeons, 15 mg/kg 3 times daily for cranes, and 25 mg/kg 3 times daily for emus would be needed to establish and maintain therapeutic tissue concentrations.

Pharmacokinetics of cephalothin and cephalexin in selected avian species.

Plasma concentrations and the biological half-lives of cephalothin and cephalexin in avian species of a variety of body sizes and metabolic rates were studied. The species chosen were eastern bobwhite quail (Colinus v virginianus), pigeons (Columba livia), hybrid rosybill ducks (Netta sp), greater sandhill cranes (Grus canadensis tabida), and emus (Dromiceius novaehollandiae). In the 1st phase of the study, cephalothin sodium was given IM in a dose of 100 mg/kg of body weight. Plasma concentrations reached peak (av 18 micrograms/ml) at 0.5 hour and were measurable 2.5 to 5.5 hours after drug administration. The biological half-life of cephalothin was 16 to 54 minutes; the half-life varied directly with increased species body weight, with the exception of the ducks studied. In the 2nd phase, cephalexin monohydrate was given orally in doses of 25, 35, and 50 mg/kg of body weight. Plasma concentrations reached peak (av 20 micrograms/ml) at 0.5 to 1 hour and were measurable 2.5 to 5.5 hours after drug administration. The biological half-life of cephalexin was 36 to 126 minutes. In the 3rd phase, differences in plasma concentrations and the half-lives of cephalexin between fed quail and fasted quail were insignificant. Dosage regimens for cephalothin of 100 mg/kg 4 times a day and for cephalexin of 35 to 50 mg/kg 4 times a day would be expected to establish and maintain therapeutic plasma concentrations in large birds (pigeons, cranes, and emus). These same doses, administered every 2 to 3 hours, would be expected to establish and maintain therapeutic plasma concentrations in smaller birds (quail, ducks).

Gentamicin tissue concentration in various avian species following recommended dosage therapy.

Plasma and tissue drug concentrations were compared in eastern bobwhite quail (Colinus virginianus virginianus) and pigeons (Columba livia) given gentamicin by IM administration at the dosage of 10 mg/kg, and in greater sandhill cranes (Grus canadensis tabida) and hybrid rosybill ducks (Netta sp) given the same antibiotic at a dosage of 5 mg/kg. Quail and cranes had significantly higher liver concentrations of gentamicin at 6 hours after injection than did pigeons and ducks. Cranes had significantly higher plasma concentrations than did ducks at 6 hours after injection. Compared with plasma values, gentamicin concentrations were significantly higher in the liver of cranes at 12 hours after injection, and in the kidneys at 18 hours.

Fiber types in the longissimus muscle from water buffalo and selected domestic beef breeds.

The distribution and area of longissimus muscle fiber types from two intact male water buffaloes, one Angus bull and one Charolais bull were evaluated. Only fibers of aerobic-oxidative capacity were present in the muscles of the water buffalo examined. The absence of αW fibers may imply some significant and distinct differences in muscle metabolic and functional characteristics. All three fibers types were present in the muscles from the Angus and Charolais bulls. These findings may implicate distinct and unique differences in meat characteristics from water buffalo, such as in organoleptic and/or processing traits.