Effect of hepatic enzyme inducers on the in vivo and in vitro metabolism of dicrotophos,dimethoate, and phosphamidon in mice

Daily 75 mg/kg phenobarbital ip injections for 3 days or 25 ppm dieldrin in the diet of mice for 14 days caused an increase in liver cytochrome P-450 and blood B-esterase. Liver A-esterase was not significantly increased. Under in vitro conditions, phenobarbital and dieldrin induced the oxidative as well as hydrolytic metabolism of dicrotophos, dimethoate, and phosphamidon by liver homogenates or combined microsomes plus 105,000g supernatant fractions. The concentration of dimethoxon was increased more than fourfold by the pretreatments after incubation for 4 hr at 37.5°C with NADPH added. The organophosphorus insecticides used in this study were not metabolized as well by the liver microsomes alone or 105,000g supernatant alone, as by the combination of microsomes and 105,000g supernatant. Under in vivo conditions in mice, phenobarbital and dieldrin treatments increased the urinary recovery of metabolites in the initial 6 hr after [¹⁴C]carbonyl-dimethoate or [¹⁴C]N-ethyl-phosphamidon administration. Analysis of urine showed that the inducers caused a more than sixfold increase in dimethoxon recovered and twofold increase in water-soluble nontoxic metabolites within 6 hr after dimethoate administration. With phosphamidon both inducers increased the rate of metabolism, and the total recovery in aqueous and chloroform fractions was decreased. These results suggest that increased dimethoate toxicity after phenobarbital and dieldrin treatments in whole animals results from stimulation of the activation of dimethoate to dimethoxon, while the increase in hydrolytic products after both pretreatments results in decreased toxicity of the direct acetylcholinesterase inhibitors, dicrotophos and phosphamidon.     

Antigenic and genotypical characterization of Newcastle disease viruses isolated in Taiwan between 1969 and 1996

Three major epidemics of Newcastle disease (ND) occurred in Taiwan over the past three decades (in 1969, 1984, and 1995). In order to gain a better understanding of the relationships between past ND epizootics in Taiwan, 36 ND viruses (NDVs) isolated between 1969 and 1996 were characterized antigenically and genotypically. The antigenicity of these viruses was analysed by their ability to cause binding of mouse monoclonal antibodies (mAbs) to cell cultures infected with the isolate. Using a panel of 22 mAbs to divide NDVs into subgroups, a total of 18 binding patterns were revealed. The sequences covering the cleavage site of the fusion protein gene of these isolates were also determined. The results of the phylogenetic analysis placed 36 NDVs into I, II, VIb, VIIa, VIII and two novel genotypes (provisionally termed X and VIh). The 1969 velogenic isolates were of genotypes X and VIh; the 1984-1985 velogenic isolates were genotyped VIb, VIh, VIIa, and X; while the 1995-1996 velogenic isolates were genotyped VIIa or VIII. Some 1969 and 1984 velogenic isolates were of the same mAbs binding pattern and genotype, and the mAbs binding patterns of the 1995-1996 isolates have not been seen before. It is concluded that velogenic NDVs of different genotype and antigenic type have co-circulated in Taiwan at least since 1969. Also there were epizootiological links between strains isolated in 1969 and 1984, whereas the 1995-1996 epidemic was caused by new antigenic variants.     

Effects of different forages on the chemical compositions and antiosteoporotic activities of velvet antlers

For this study, we aimed to assess the dose–response antiosteoporotic effects of the middle section of velvet antlers (VAs) from sika deers (Cervus nippon) fed with different types of fodders. VAs prepared from farmed sika deers fed with feed mixtures containing sorghum distillery residue (VA‐SDR) or without SDR (SDR replaced with hay, VA‐Hay) were divided into upper (VAU), middle (VAM) and basal (VAB) sections. The chemical constituents of the middle sections obtained from each VA type were compared, and their antiosteoporotic activities were evaluated using rats with ovaries removed surgically (ovariectomy, OVX). The VA‐Hay exhibited markedly increased iron and cysteine levels, whereas the VA‐SDR exhibited markedly increased level of alcoholic extract and testosterone. Both VA‐Hay‐ and VA‐SDR‐treated rats exhibited increased femur strength compared with the control group. However, VA‐SDR exhibited greater bone‐strengthening effects than did VA‐Hay. The serum osteocalcin and estradiol levels were significantly moderated in the VA‐Hay group alone. These results suggest that VA‐SDR and VA‐Hay prevent the loss of bone strength, and preserve trabecular architecture connectivity in an estrogen‐deficient state. However, differences in the chemical compositions of different forages may be responsible for the varying antiosteoporotic mechanisms observed. Thus, the addition of SDR in deer forage may enhance antiosteoporosis activity in VAs, and confer considerable economic and ecological benefits.     

Expression profiles of immunity and reproductive genes during transition period in Holstein cattle

The transition period is a critical time for dairy cows as the animal is subjected to the physiological stress accompanying parturition. Immunosuppression and health status were examined during this period in 80 Holstein cows. Blood samples were taken from each cow 3, 2 and 1 week before and after calving, and at calving (0 day). RNA was extracted and subjected to real‐time PCR to determine mRNA levels for the immune‐related genes TLR 2, 4, 6, 7 and β‐defensin 5 in addition to the reproduction‐related genes prolactin and IGF‐I. Results showed significant up‐regulation of pro‐inflammatory‐selected genes, TLR 4, 6 7 and β‐defensin 5 at the third‐week post‐calving; however, earlier periods had lower expression of such genes. In contrast, the immunosuppression biomarker TLR2 gene was up‐regulated at calving and 1 week after parturition and then down‐regulated again at second and third week. Prolactin and IGF‐I genes expression levels were significantly and gradually increased mainly post‐partum. This research highlights that the expression patterns of TLRs, BNBD5, PRL and IGF‐I could be biomarkers to follow up immune and reproductive status of dairy cow at peri‐parturient period to predict the most susceptible risk time for disease incidence and to build up management protocol.     

Can we see fade? A survey of anesthesia providers and our ability to detect partial neuromuscular block in dogs

Objective

To assess the ability to visually detect fade during train-of-four (TOF) or double burst stimulation (DBS) in anesthetized dogs recovering from nondepolarizing neuromuscular block.

Multiple‐Particle Tracking Study of Microheterogeneity of Nutrim‐10 Suspensions

Nutrim is a newly developed food product containing dietary soluble fiber β‐glucan. The microstructural heterogeneities of Nutrim‐10 suspensions were investigated by monitoring the thermally driven displacements of well‐dispersed microspheres through video fluorescence microscopy. By comparing the distribution of the time‐dependent mean‐square displacement (MSD) of polystyrene microspheres embedded in three concentrations of Nutrim‐10 suspensions, we found that the degree of heterogeneity of suspensions increased dramatically within a narrow range of Nutrim‐10 concentrations. The ensemble‐averaged MSD of 5.5% Nutrim‐10 suspension exhibited a power‐law behavior scaling linearly with time, which was similar to the behavior for a homogeneous aqueous glycerol solution. But the MSD distribution was wider and more asymmetric than for glycerol. Increasing Nutrim‐10 concentration rendered the MSD distribution much more asymmetric and skewed. This study provided a quantitative method to characterize the organization of Nutrim‐10 in suspension.     

Expression, purification, and characterization of the maltooligosyltrehalose trehalohydrolase from the thermophilic archaeon Sulfolobus solfataricus ATCC 35092

The maltooligosyltrehalose trehalohydrolase (MTHase) mainly cleaves the alpha-1,4-glucosidic linkage next to the alpha-1,1-linked terminal disaccharide of maltooligosyltrehalose to produce trehalose and the maltooligosaccharide with lower molecular mass. In this study, the treZ gene encoding MTHase was PCR-cloned from Sulfolobus solfataricus ATCC 35092 and then expressed in Escherichia coli. A high yield of the active wild-type MTHase, 13300 units/g of wet cells, was obtained in the absence of IPTG induction. Wild-type MTHase was purified sequentially using heat treatment, nucleic acid precipitation, and ion-exchange chromatography. The purified wild-type MTHase showed an apparent optimal pH of 5 and an optimal temperature at 85 degrees C. The enzyme was stable at pH values ranging from 3.5 to 11, and the activity was fully retained after a 2-h incubation at 45-85 degrees C. The k(cat) values of the enzyme for hydrolysis of maltooligosyltrehaloses with degree of polymerization (DP) 4-7 were 193, 1030, 1190, and 1230 s(-1), respectively, whereas the k(cat) values for glucose formation during hydrolysis of DP 4-7 maltooligosaccharides were 5.49, 17.7, 18.2, and 6.01 s(-1), respectively. The K(M) values of the enzyme for hydrolysis of DP 4-7 maltooligosyltrehaloses and those for maltooligosaccharides are similar at the same corresponding DPs. These results suggest that this MTHase could be used to produce trehalose at high temperatures.     

Pharmacokinetics of (-)-epigallocatechin-3-gallate in conscious and freely moving rats and its brain regional distribution

A liquid chromatography technique coupled with tandem mass spectrometry (LC-MS/MS) electrospray ionization was used to measure (-)-epigallocatechin-3-gallate (EGCG) in rat plasma. This method was applied to investigate the pharmacokinetics of EGCG in a conscious and freely moving rat by an automated blood sampling device. Multiple reaction monitoring (MRM) was used to monitor the transition of the deprotonated molecule m/z of 457 [M - H]- to the product ion 169 for EGCG and the m/z of 187 to 164 for the internal standard. The limit of quantification (LOQ) of EGCG in rat plasma was determined to be 5 ng/mL, and the linear range was 5-5000 ng/mL. The protein binding of EGCG in rat plasma was 92.4 +/- 2.5%. The brain distribution result indicated that EGCG may potentially penetrate through the blood-brain barrier at a lower rate. The disposition of EGCG in the rat blood was fitted well by the two-compartmental model after intravenous administration (10 mg/kg, iv). The elimination half-life of EGCG was 62 +/- 11 and 48 +/- 13 min for intravenous (10 mg/kg) and oral (100 mg/kg) administration, respectively. The pharmacokinetic data indicate that the oral bioavailability of EGCG in a conscious and freely moving rat was about 4.95%.