Analysis of protein induction in the CNS of SJL mice with experimental allergic encephalomyelitis by proteomic screening and immunohistochemistry.

Experimental allergic encephalomyelitis (EAE) is an autoimmune disease characterized by demyelination and inflammatory infiltrates in the CNS, and it is an animal model of multiple sclerosis. Piperonyl butoxide (PBO) suppresses disease in EAE mice, and it exhibits a dual effect on cytochrome P450s that manifests in a transient inhibitory phase followed by induction. In order to identify the expression of proteins associated with EAE, a proteomic screening was performed on hindbrain microsomes from control + vehicle, control + PBO, EAE + vehicle, and EAE + PBO female mice. Glucose regulated protein 94 (Grp94) and coagulation factor VIII were among the proteins identified in EAE + vehicle and EAE + PBO mice. Immunohistochemical staining of Grp94 was present in some neurons and oligodendrocytes in hindbrain sections from control animals, and in some cells within inflammatory infiltrates in EAE animals. Since Grp94 (also known as Gp96) can partake in antigen presentation and induction of proinflammatory cytokine expression, its presence in these cells suggests that it may play a role in the pathogenesis of EAE. Coagulation factor VIII is carried and protected by von Willebrand factor. Immunohistochemical staining of von Willebrand factor revealed its presence in some vessels within hindbrain sections from control animals. In EAE animals, the number of labeled vessels was significantly increased, and extracellular granular deposits were observed around labeled vessels indicating that the breakdown of the blood-brain barrier that occurs in EAE permitted its extravasation into the CNS. Additional proteins were identified in the different groups of mice by proteomic screening, but confirmation of their expression profile awaits investigations by independent measures.     

Role of T-lymphocyte subsets in recovery from herpes simplex virus infection.

Our investigations probed the nature of different T-lymphocyte subsets effecting clearance of herpes simplex virus after infection of the pinna. Cell populations from animals recently infected subcutaneously or intraperitoneally (acute population) or from animals infected 6 weeks previously (primed population) or the latter cells reimmunized in vitro with virus (memory population) were studied. Viral clearance was a function of the Lyt 1+2- subset in the acute population, but with the memory population both Lyt 1+ and Lyt 2+ cells affected clearance. In primed populations, viral clearance was effected only by the Lyt 2+ subset. The ability of the various cell populations to adoptively transfer delayed-type hypersensitivity was also studied. Only acute population cells from animals infected subcutaneously and memory population cells transferred delayed-type hypersensitivity. In both cases, the cell subtype was Lyt 1+2-. Our results demonstrated that the delayed-type hypersensitivity response does not always correlate with immunity to herpes simplex virus. Multiple subsets of T cells participate in viral clearance, and their respective importances vary according to the stage of the virus-host interaction.     

Lipopolysaccharide treatment modifies pH- and cation-dependent ecto-ATPase activity of endothelial cells.

The aim of this study was to investigate the distribution pattern of Ca2+- and Mg2+-dependent ecto-ATPases on the surface of rat brain capillary endothelial cells (ECs) in control and lipopolysaccharide (LPS)-treated animals. Ecto-ATPases in the membrane of vascular endothelial cells are suggested to play a crucial role in thromboregulation. Loss of this enzyme activity after oxidative stress and upregulation of the enzyme chain hydrolyzing extracellular ATP after transient forebrain ischemia have also been reported. We used histochemistry to localize the activities of this enzyme on ECs and found pH- and cation-dependent changes in the localization of enzyme activity both in control and in LPS-treated animals. These findings suggest the presence of more than one ecto-ATPase enzyme on the surface of rat capillary ECs. The different behavior of ECs after LPS treatment is the target of further investigations. The increased ecto-nucleotidase activity might play a role in nucleotide-mediated cellular responses after bacterial infections.     

Effects of sedation on the controlled pulmonary circulation

The pulmonary circulatory effects of sedatives and groups of sedatives in dosages commonly employed as pre cardiac catheterization sedatives were evaluated. The sedatives included: meperidine, thiamylal, hydroxyzine, promethazine, meperidine + chlorpromazine + promethazine and meperidine + hydroxyzine. An animal preparation with constant pulmonary blood flow and left atrial pressure was employed. This preparation has the unique advantage of permitting measurement of the active pulmonary vascular resistance change since passive effects are controlled. In order to establish the significance of a given change in active PVR, a dose response relationship for serotonin was established and the effect of the sedatives was calibrated against it. PVR increased following infusion of all sedatives tested except hydroxyzine and chlorpromazine.A technique was detailed to permit subtraction of passive PVR change from total PVR change in intact, unsedated animals. Results of active change in PVR of previous investigations in intact, unsedated animals were compared to the active change observed in this experiment and were found to be similar in direction and magnitude. This investigation stresses the need to consider possible effects of sedatives on the pulmonary circulation.     

Effects of repetitive immunogen injections and fasting versus feeding on iron,zinc, and copper metabolism in chicks

Most investigations on the effect of immunogenic challenge on trace-mineral metabolism use a single immunogen injection in fasted animals. Because these investigations are not representative of realistic situations in which animals are constantly exposed to immunogens and still consume feed, the following studies were done. In Expt. 1, chicks were injected ip with sheep red blood cells (SRBC), Sephadex, SRBC+Sephadex, or saline. Chicks were then fasted or fed equal amounts of feed (equal fed) for 16 h. Immunogen injection decreased serum Fe and Zn and increased serum Cu within each feeding program. Differences in Cu, and to a lesser extent Zn, concentrations between immunogen- and saline-injected chicks were more pronounced in fasted than in equal-fed chicks. In Expt. 2, equalfed chicks were injected ip every 48 h for 6 d with SRBC+Sephadex or saline. Two days after each injection, tissues were taken. An additional group of chicks was injected once and subsequently fasted 16 h, whereupon tissues were taken. Changes in plasma Fe, Zn, Cu, and ceruloplasmin; hepatic Fe, Zn, Cu, and metallothionein; pancreatic Fe and Zn; and splenic Fe in repetitively injected chicks were different from changes observed in chicks injected once. The results indicate that the trace-mineral response to immunogenic challenge is dependent upon the number of immunogen injections and the nutritional state of the animal.     

Alternative ultrastructural localization of Dolichos biflorus lectin binding sites in proton secreting parietal cells of mice

High amount of N-acetyl-D-galactosamine specific lectin binding sites were detected on the canalicular membranes of human parietal cells. Our present model investigations on mice showed that the intracellular distribution of the terminal N-acetyl-D-galactosamine containing glycoprotein highly depends on the actual functional state of the parietal cells. In the normal gastric mucosa 40%-60% of parietal cells react positively after staining with horseradish peroxidase or biotin labelled Dolichos biflorus lectin. Ultrastructurally lectin binding sites occur mainly on the basolateral membrane infoldings in fed animals, while they are present exclusively on the canalicular membranes of fasting mice, suggesting that the alternative appearance of lectin binding sites on the opposite membrane areas of parietal cells is tightly coupled to their main function, to H+ secretion.     

The uptake and accumulation of iron by the intestinal bacterium Desulfotomaculum acetoxidans DSM 771

Sulfate-reducing bacteria (e.g. Desulfotomaculum acetoxidans) exist in animal intestine. These bacteria are able to bind heavy metals (e.g. cadmium or lead). Comparative investigations on the composition of cellular walls of Desulfotomaculum acetoxidans--depending on the initial Fe2+ supplement in the medium (7.5, 57.5 and 507.5 M) were performed. Iron(II) was cumulated as FeS or as pyrite (FeS2). However, if the initial amount of iron was higher, its majority (46% 85%) was transported onto the membrane. It was determined that the siderophore found in Desulfotomaculum acetoxidans was deferroxamine as in animals.     

Reduction of the RT6.2+ subset of T lymphocytes in brown Norway rats with mercury-induced renal autoimmunity

Chemically induced autoimmunity is a recently recognized environmental hazard that may affect individuals genetically predisposed to autoimmune disease and chronically exposed to certain chemicals. For example, moderate concentrations of mercury may lead to renal autoimmune disease in a small but significant percentage of the exposed population. Mercury also induces autoimmune glomerulonephritis in susceptible Brown Norway (BN) and MAXX inbred strain rats. Autoimmune responses, directed to epitopes of the renal glomerular basement membrane (GBM), are rapid in onset and have a self-limiting course in mercury-treated rats. Both regulatory T cells and idiotype-anti-idiotype network have been implicated in the resolution of this autoimmune process. In our investigations of immune regulation of mercury-induced autoimmune glomerulonephritis, we have used flow cytometry to quantitate lymphocyte subpopulations in the spleen and lymph nodes of mercury-treated and control BN rats. Of particular interest was the RT6+ T cell subset, that appears to have important immunoregulatory properties in a rat model of autoimmune insulin-dependent diabetes mellitus. Spleen and lymph nodes from control BN rats contained 22 and 52%, respectively, RT6+ cells. Spleens from mercury-treated animals contained 21% RT6+ cells on Day 10 of treatment, 13% on Day 17, 16% on Day 24 and 20% on Day 30. Lymph nodes from the same rats had 36% RT6+ cells on Day 10, 23% on Day 17, 29% on Day 24, and 28% on Day 30. The decrease in RT6+ cells correlated inversely with autoimmune responses to GBM, which peaked on Days 17-24 and declined by Day 30. Moreover, autoimmune responses were also associated with elevated RT6-:RT6+ T cell ratios. Similar results were obtained in two additional groups of BN rats, comprising both younger and older animals, sacrificed at Day 18 of mercury treatment. Analysis of other lymphocyte subpopulations demonstrated a decrease of CD4+ and CD5+ cells, whereas B cells as well as CD8+, IL-2 receptor+, and MHC class II+ subsets showed no consistent correlation with the onset or resolution of the autoimmune process. These findings suggest that mercury-induced changes in RT6+ T lymphocytes may be related to the development of renal autoimmune disease in genetically predisposed BN rats.     

黄河三角洲土壤运行多样性初步调查研究

1995－1996年对黄河三角洲9个代表性样区,进行了土壤运动定性,定量调查,并对古代,近代,现代黄河三角洲3个代表性样区开展了土壤动物周年调查,研究了土壤动物的种类组成,分布和节季变化,以及土壤动物多样性,调查研究结果,黄河三角洲共鉴定出土壤运动38种,土壤运动种类组成和数量分布因三个三角洲成土年龄不同而存在差异,土壤运动多样性以夏季为丰富,黄河三角洲土壤运动多样性]与均匀度表现为正相关关系,与单纯度呈负相关趋势。     

Resistance to antibiotics in humans and animals on the Cape Verde Islands

From the Cape Verde Islands no studies are known about an efficient treatment with antibiotics and chemotherapeutics. Because of this human samples and samples of domestic animals were collected, the different bacterias were isolated and the antibiograms were analyzed by agar-diffusions-test. The human samples consisted of wound-, eye-, anal-, vaginal- and throat-smears, the animal samples consisted of jaw-, anal- and wound-smears. the aim of these investigations was to make efficient treatment with antibiotics and chemotherapeutics possible. The best efficacy had Cefotaxim, Gentamicin and Trimethoprim + Sulfonamid. Very poor were Penicillin G, Aminopenicillin and Cephalexin.     

The effect of body weight on the degree of blood velocity profile skewness in the aortic annulus in domestic pigs

We investigated the blood velocity profile in the aortic annulus (AA) in two groups of domestic pigs using epicardial Doppler echocardiography. The velocity profile skewness in terms of max/mean TVI (the ratio of maximal to cross-sectional mean time-velocity integral along the diameter) was 1.107 +/- 0.01 in the small pigs (n = 10; body weight: 24.6 +/- 0.8 kg) and 1.216 +/- 0.026 in the large pigs (n = 8; body weight: 50.6 +/- 2.5 kg) (P = 0.002). The velocity profile in the AA is more skewed in large animals than in small animals and the skewness in the larger animals is similar to that in normal adult humans. This study shows the importance of choosing animals of sufficient size if flow method investigations are to be performed. This is particularly important for ultrasound Doppler investigations based on a limited sample of velocities across the flow channel.     

Reduction in animal numbers by long-term implantation of intravenous and intra-arterial catheters in thyroparathyroidectomized rats

The thyroparathyroidectomized (TPTx) rat has been extensively used to study parathyroid hormone (PTH)-mediated bone resorption by measuring systemic Ca2+ concentrations. Animals have been traditionally used acutely; that is, they are often infused immediately after surgery and are sacrificed after a single use. To perform multiple experiments using a single group of animals we developed a system of long-term implanted intravenous/arterial catheters. Using calcitonin (CT) as a positive control, we successfully completed 12 separate controlled subexperiments documenting significant reductions in PTH-induced hypercalcemia in rats of the CT group. We then successfully completed two separate TPTx subexperiments, using a 3 x 3 Latin square experimental design. In both subexperiments, CT significantly inhibited the increase of blood Ca2+ concentration resulting from continuous PTH infusion. Our results indicate that, by combining the long-term use of catheters with the Latin square design, we can successfully reduce the number of animals used, increase the number of compounds screened, and improve the quality of the data. Although results of this study confirmed the acceptability of multiple infusions in anti-resorptive studies, investigations into the applicability of this set up to other areas of study requiring infusions and frequent blood sample collections seem appropriate.     

The adaptive strategy of rodent populations living in conditions of radioactive and chemical environmental pollution

The comparative analysis of demographic, morphological and physiological processes in mouselike rodents in pollution zones (90Sr + 90Y, 137Cs) on East-Ural radioactive track (EURT) and (Cu + Cd + Pb + Zn + SO2) on a site near copper-smelting factory is carried out. The direct (not mediated) defeat of animals by an irradiation leads to inherited adaptation (density preservation, tolerance increase to pollution, migration decrease and so forth). The mediated defeat of animals at pollution by metals influences animals as a result of degradation of a vegetative cover, reducing a forage reserve, shelters and reproduction places. Population is decreasing, migration is increasing. Hence, population reacts onto direct defeat of animals or on inhabitancy locuses degradation, id est unspecifically, without dependence from the physical and chemical nature of pollution.     

COMPARATIVE STUDIES ON RESPIRATION : I. INTRODUCTION.

A series of investigations on respiration with improved quantitative methods has been commenced. The first of these are here described. They show that when anesthetics are employed in sufficient concentration to produce any result, plants show a rise in the rate of respiration which is followed by a fall. In the animals studied, the rise (found in higher concentrations only) was preceded by a temporary fall which is not entirely due to lowering of muscular activity or tonus. In lower concentrations the effect on animals was merely a decrease of respiration. The results of all the investigations are opposed to the theory of Verworn.     

Successful transfer of vitrified Ilama (Lama glama) embryos

The exploitation of the domestic animals species of South American camelids is of great social importance for the native people living in the High Andes. The reproductive physiology of these species is a unique challenge in the development of advanced breeding techniques. At present, the cryopreservation of embryos has not been developed and very few investigations have been conducted. The objective of the present work was to evaluate the in vivo survival of vitrified llama embryos after transfer to recipient females. Donors females were treated with a CIDR-estradiol benzoate-eCG regimen and were mated naturally 6 days after CIDR withdrawal. One ovulatory dose (8 microg) of GnRH was administered immediately after mating. A second mating was allowed 24 h later. Embryo recovery was performed nonsurgically between 8 and 8.5 days after the first mating. Twenty-two ova/embryos were recovered from 12 donor females. Hatched blastocysts were exposed to vitrification solution (20% glycerol + 20% ethylene glycol + 0.3 M sucrose + 0.375 M glucose + 3% polyethylene glycol (P/V)) in three steps, and after loading into 0.25 ml straws, were plunged into liquid nitrogen. For embryo transfer, recipients animals were ovulation-synchronized using GnRH administered at the same time as donors. A total of eight vitrified-warmed embryos and 12 fresh embryos were nonsurgically transferred to four and six recipient females, respectively (two embryo per recipient). The pregnancy rates were 50 and 33.3% for recipients that had received vitrified embryos and fresh embryos, respectively. The results demonstrated the effectiveness of this simple vitrification method for cryopreservation of llama embryos.     

Chronopharmacologic characteristics of the development of tolerance for the narcotic effect of ethanol in a hot climate

The experiments have been carried out on rats tested by Parsolt in various time of the day in January and July. The results of investigations showed that the behaviour activity of animals was exposed to daily and season fluctuations. The tolerance development to the ethanol narcotic effect depends upon the behaviour activity of animals. During summer the increasing of ethanol toxicity was observed. In highly active rats as distinct from low active animals the real increasing of noradrenaline concentration and the decreasing of dofamine concentration were noted. Ethanol changes the correlations of blood monoamines and the behaviour activity of animals.     

Genome Mapping in Aquatic Animals: Progress and Future Perspectives

The genome of aquatic animals is poorly understood and information from different taxonomic groups is sketchy. While there have been intensive genomic studies on some fish models, investigations on other fishes and invertebrates have been scarce. Yet there are recently some coordinated studies on genome mapping in a number of aquaculture animals of economic importance. This review summarizes information available on genome mapping of the important fish models and aquaculture animals. The future perspectives of this field of studies are discussed.     

Alternative ultrastructural localization of Dolichos biflorus lectin binding sites in proton secreting parietal cells of mice

Summary High amount of N-acetyl-d-galactosamine specific lectin binding sites were detected on the canalicular membranes of human parietal cells. Our present model investigations on mice showed that the intracellular distribution of the terminal N-acetyl-d-galactosamine containing glycoprotein highly depends on the actual functional state of the parietal cells. In the normal gastric mucosa 40%–60% of parictal cells react positively after staining with horseradish peroxidase or biotin labelled Dolichos biflorus lectin. Ultrastructurally lectin binding sites occur mainly on the basolateral membrane infoldings in fed animals, while they are present exclusively on the canalicular membranes of fasting mice, suggesting that the alternative appearance of lectin binding sites on the opposite membrane areas of parietal cells is tightly coupled to their main function, to H⁺ secretion.     

Genome mapping in aquatic animals: progress and future perspectives

The genome of aquatic animals is poorly understood and information from different taxonomic groups is sketchy. While there have been intensive genomic studies on some fish models, investigations on other fishes and invertebrates have been scarce. Yet there are recently some coordinated studies on genome mapping in a number of aquaculture animals of economic importance. This review summarizes information available on genome mapping of the important fish models and aquaculture animals. The future perspectives of this field of studies are discussed.     

Compendium of the antidiabetic effects of supranutritional selenate doses. In vivo and in vitro investigations with type II diabetic db/db mice

In recent years, a number of investigations on the antidiabetic effects of supranutritional selenate doses have been carried out. Selenate (selenium oxidation state +VI) was shown to possess regulatory effects on glycolysis, gluconeogenesis and fatty acid metabolism, metabolic pathways which are disturbed in diabetic disorders. An enhanced phosphorylation of single components of the insulin signalling pathway could be shown to be one molecular mechanism responsible for the insulinomimetic properties of selenate. In type II diabetic animals, a reduction of insulin resistance could be shown as an outcome of selenate treatment. The present study with db/db mice was performed to investigate the antidiabetic mechanisms of selenate in type II diabetic animals. Twenty-one young adult female db/db mice were randomly assigned to three experimental groups (selenium deficient=0Se, selenite-treated group=SeIV and selenate-treated group=SeVI) with seven animals each. Mice of all groups were fed a selenium-deficient diet for 8 weeks. The animals of the groups SeIV and SeVI were supplemented with increasing amounts of sodium selenite or sodium selenate up to 35% of the LD50 in week 8 in addition to the diet by tube feeding. Selenate treatment reduced insulin resistance significantly and reduced the activity of liver cytosolic protein tyrosine phosphatases (PTPs) as negative regulators of insulin signalling by about 50%. In an in vitro inhibition test selenate (oxidation state +VI) per se did not inhibit PTP activity. In this test, however, selenium compounds of the oxidation state +IV were found to be the actual inhibitors of PTP activity. Selenate administration in vivo further led to characteristic changes in the selenium-dependent redox system, which could be mimicked in an in vitro assay and provided further evidence for the intermediary formation of SeIV metabolites. The expression of peroxisome proliferator-activated receptor gamma (PPARgamma), another important factor in the context of insulin resistance and lipid metabolism, was significantly increased by selenate application. In particular, liver gluconeogenesis and lipid metabolism were influenced strongly by selenate treatment. In conclusion, our results showed that supranutritional selenate doses influenced two important mechanisms involved in insulin-resistant diabetes, namely, PTPs and PPARgamma, which, in turn, can be assumed as being responsible for the changes in intermediary metabolism, e.g., gluconeogenesis and lipid metabolism. The initiation of these mechanisms thereby seems to be coupled to the intermediary formation of the selenium oxidation state +IV (selenite state) from selenate.