Subchronic oral toxicity (six months) of carboxyethylgermanium sesquioxide [(HOOCCH2CH2Ge)2O3]n in rats

After briefly renewing toxicological data on germanium compounds, the authors report on the subchronic oral toxicity of carboxyethylgermanium sesquioxide in rats. During six months, male and female animals received 1 g kg^?1. day^?1. No particular toxic symptoms, and no behaviour problems except a small decrease of body weight in male rats, at the end of the six-month experimentation period, were observed. A significant decrease of erythropoiesis and some significant changes in leucocyte ratios were demonstrated. The main marked effect was a moderate renal dysfunction characterized by a tubular disease with the presence of cylinders, swelling of tubulus cells and flocculus deposits. Germanium urinary excretion was constant and linked to the received dose. Six months later, no preferential accumulation in organs was evident.     

Resolution of lymphocyte subpopulations derived from rat spleen by polyamine-graft-PHEMA copolymer

The retention behavior of lymphocyte subpopulations, B cell, T cell and null cell, derived from rat spleen to polyamine-graft-poly(2-hydroxyethyl methacrylate) copolymer (HA) surface was investigated, focusing on the conformational transition of the polyamine side chain as well as the protonation of amino groups in the polyamine grafts. Furthermore, the availability of HA was discussed as a column adsorbent for separation of lymphocyte subpopulations derived from spleen. The conformational transition of polyamine grafts significantly influenced the mode of retention of lymphocyte subpopulations. When polyamine grafts existed in an aggregated conformation (protonatin degree α < 0.5), the retention of lymphocyte subpopulations was decreased in the order B cell> null cell> T cell. On the other hand, when polyamine existed in an extended conformation into the aqueous interior from the matrix interface (α > 0.5), T cell retention became greater than null cell retention, resulting in a decreased B cell> T cell> null cell order. These results indicate that the differential retention of spleen lymphocyte subpopulations is attributed to their differential responses to the change in matrix interface accompanied by the protonation of amino groups. Furthermore, spleen lymphocytes were compared with lymph node lymphocytes in terms of resolution efficacy by an HA copolymer column.     

大鼠脑组织中不同部位单胺类神经递质含量的HPLC测定

单胺类神经递质,多巴胺(DA)、去甲肾上腺素(NA)和肾上腺素(A)是哺乳动物和人类中枢重要的信息传递物质。了解脑组织中各部位单胺类神经递质的含量对于研究其生理功能、有关疾病的诊断和药物及手术疗效的评价均有重要的意义。     

Determination of levodopa methyl ester and its metabolites in rat serum by CZE with amperometric detection

A reliable and reproducible method, capillary zone electrophoresis with amperometric detection (CZE–AD), has been developed for separation and quantification of levodopa methyl ester (LDME) and its biotransformation products levodopa (L-DOPA) and dopamine (DA) in rat serum. A carbon-disk electrode was used as working electrode. The optimum conditions for CZE detection were 50 mmol L^–1 phosphate solution at pH 7.0 as running buffer, 17 kV as separation voltage, 1.0 V (vs Ag/AgCl, 3.0 mol L^–1) as detection potential, and sample injection for 8 s at 17 kV. The linear ranges were from 2.4×10^–2 to 2.2 g mL^–1 for LDME, 2.9×10^–1 to 49.5 g mL^–1 for L-DOPA, and 1.4×10^–2 to 1.5 g mL^–1 for DA with correlation coefficients of 0.9997, 0.9994, and 0.9999, respectively. The detection limits for LDME, L-DOPA, and DA were 14.6, 98.0, and 9.7 ng mL^–1, respectively. Recoveries were 80.3% for LDME, 93.5% for L-DOPA, and 86.5% for DA. This method was applied to serum samples after intravenous injection of LDME and L-DOPA to rats.     

Nuclear Magnetic Resonance-Based Blood Metabolic Profiles of Rats Exposed to Short-Term Caloric Restriction

Caloric restriction increases the life-span of a number of organisms. The relationship between the increase in life-span and the extent of caloric restriction, however, varies among species. The underlying mechanisms are yet unknown, but appear to be related to changes in metabolism. In order to investigate the metabolic response of caloric restriction of rats, here is presented the first nuclear magnetic resonance (NMR) spectroscopy-based study of how blood metabolite profiles are influenced by graded levels of caloric restriction. The study involved three groups of obese rats exposed to 0, 20, and 40 percent caloric restriction for five days. Blood serum from each individual was analyzed by ¹H NMR and the resulting spectra were subjected to multivariate analysis by unsupervised principal component analysis and supervised orthogonal-partial least square discriminant analysis. The analyses revealed that a response to caloric restriction was present at 20 percent caloric restriction. The metabolites that distinguished the profiles at 20 percent restriction deviated from those at 40 percent restriction. The changes induced by caloric restriction were most clearly observed as an increased level of 3-hydroxybutyrate, and decreased levels of lipids and pyruvate. The metabolic responses of rats exposed to caloric restriction are in good agreement with a switch in metabolism from anabolic pathways towards fatty acid catabolism and gluconeogenesis, which is consistent with previous observations for mice.     

The effect of 2.1 T static magnetic field on astrocyte viability and morphology

The viability and a number of morphological properties of in situ astrocytes of rat spinal cord cultures including changes in surface area and migration of both cell body and nucleus were investigated at magnetic field intensities comparable to those currently used for magnetic resonance imaging. Viability of rat spinal astrocytes was studied after up to 72 hours of 2.1T static magnetic field exposure. Surface areas and two-dimensional centroids of both soma and nucleus after 2 hours of magnetic field exposure were determined and compared with those of the same cells before magnetic field exposure. Cell membrane ruffling was quantified using fractal analysis.     

Ultra‐fast LC–ESI‐MS/MS method for the simultaneous determination of six highly toxic Aconitum alkaloids from Aconiti kusnezoffii radix in rat plasma and its application to a pharmacokinetic study

A fast, sensitive, and efficient ultra‐fast LC–ESI‐MS/MS method was developed for the simultaneous quantitation of six highly toxic Aconitum alkaloids, that is, aconitine, mesaconitine, hypaconitine, benzoylaconine, benzoylmesaconine, and benzoylhypaconine, in rat plasma after oral administration of crude ethanol extracts from Aconiti kusnezoffii radix by ultrasonic extraction, reflux extraction for 1 h, and reflux extraction for 3 h, respectively. The separation of six Aconitum alkaloids and aminopyrine (internal standard) was performed on an InertSustain® C₁₈ column, and the quantification of the analytes was performed on a 4000Q ultra‐fast LC–MS/MS system with turbo ion spray source in the positive ion and multiple‐reaction monitoring mode. Absolute recoveries ranged within 65.06–85.1% for plasma samples. The intra‐ and interday precision and accuracy of analytes were satisfactory. The methods were validated with sensitivity reaching the lower LOQ for aconitine, mesaconitine, hypaconitine, benzoylaconine, benzoylmesaconine, and benzoylhypaconine, which were 0.025, 0.025, 0.050, 0.025, 0.025, and 0.100 ng/mL, respectively. The method was successfully applied to a pharmacokinetic study of six Aconitum alkaloids in rat plasma after oral administration of crude ethanol extracts from the raw root of Aconitum kusnezoffii Reichb. by three different extraction processes.     

Screening and analysis of metabolites in rat urine after oral administration of Apocynum venetum L. extracts using HPLC–TOF‐MS

HPLC with diode array detection and ESI‐TOF‐MS was used for the study of the constituents in Apocynum venetum L. extracts and the metabolites in rat urine after oral administration of A. venetum L. extracts. A formula database of the known constituents in A. venetum L. was established, and 21 constituents were rapidly identified by accurately matching their molecular masses with the formulae of the compounds in the database. Furthermore, 34 metabolites were detected and elucidated in the rat urine. The scientific and plausible biotransformation pathways of the flavonoid components in A. venetum L. were also proposed together with the presentation of clues for potential mechanisms of bioactivity. This specific and sensitive HPLC–ESI‐TOF‐MS method can be used to identify the chemical components in the extracts of A. venetum L. and their metabolites in rat urine. This method can also be used to reveal the possible metabolic mechanisms of action of the extract components in vivo.     

Dissociation between lactate accumulation and acidosis in middle cerebral artery-occluded rats assessed by P and H NMR metabolic images under A 2-T magnetic field

The relationships among tissue edema, lactate accumulation, and intracellular pH in middle cerebral artery (MCA)-occluded rats were investigated with multiecho ¹H magnetic resonance imaging and spatially resolved metabolic images constructed by ¹H and ³¹P nuclear magnetic resonance (NMR) chemical shift imaging (CSI). For the effective and sensitive detection of NMR signals from the brain, outer volume suppression (OVS), reduced k-space sampling and proton irradiation were incorporated into the CSI sequences. The consecutive three measurements of calculated T₂ image, lactate image, and pH image which were required for 3.75 h were repeated for four cycles of 1–16 h after MCA occlusion. Tissue edema and lactate accumulation in the infarcted region were gradually and consistently increased during the 15-h observation period. In contrast, severe acidosis was already detected on the first pH image (2–4.7 h after MCA occlusion); thereafter, the degree of acidosis became milder and showed no further progression. The dissociation between the time courses of the lactate accumulation and pH decrease was clearly demonstrated by the NMR metabolic images. Acid-base balance in cerebral infarction might be affected not only by lactate production but also by complicated interactions with tissue edema and some other factors.     

The rheological properties of the arterial wall

Key words The rheological properties of rat arteries were studied in vitro. Using intact arterial segments undergoing large deformations in longitudinal and circumferential direction, both the stress-strain relationship and the corresponding histological changes with deformation are presented. Correlations between the mechanical properties and the histological structure are discussed.