Hepatic collagen production in the rat is unaffected by methotrexate

Methotrexate (MTX) has been implicated in the pathogenesis of hepatic fibrosis. However, no information exists regarding the effects of MTX on hepatic collagen metabolism. Therefore, we studied the role of MTX in hepatic collagen production in vivo in rats receiving an 8-week course of varying doses of MTX. Twenty-four hours prior to sacrifice animals received an injection of [5-3H]proline. Collagen was extracted with hot trichloroacetic acid and the proteinbound [3H]hydroxyproline was used as a measure of de novo collagen production. The hepatic collagen content was essentially the same in the control and treatment groups in spite of evidence of hepatotoxicity. Similarly, no significant differences were present among the control and MTX-treated groups in the de novo absolute collagen production. In summary, we found no evidence of increased hepatic fibrogenesis in small groups of animals after 8 weeks of treatment with MTX. Data clearly supporting the claim that MTX itself is responsible for hepatic fibrosis are lacking.     

The effect of migration on metapopulation stability is qualitatively unaffected by demographic and spatial heterogeneity

Coupled map lattices (CMLs), using two coupled logistic equations, have been extensively used to model the dynamics of two-patch ecological systems. Such studies have revealed that migration rate plays an important role in determining the dynamics of the system, particularly when the two maps differ in their intrinsic growth rate parameter, r. However, under more realistic assumptions, a metapopulation can be expected to consist of more than two subpopulations, each with its own demographic parameters, which will in part be a function of the environment of that patch. The role of the spatial arrangement of heterogeneous (i.e. with different r values) subpopulations in shaping the dynamics of such a metapopulation has rarely been investigated. Here, we study the effect of demographic and spatial heterogeneity on the stability of one- and two-dimensional systems of 64 coupled Ricker maps with different r values, under periodic and absorbing boundary conditions. We show that the effects of migration rate on metapopulation stability do not depend upon either the precise spatial arrangement of the subpopulations in the lattice, or on the presence of a moderate proportion of vacant (uninhabitable) patches in the lattice. The results, thus, suggest that metapopulation models are robust to variation in spatial arrangement of patch quality and, hence, of demographic parameters. We also show that for any given arrangement of the patches, maximum stability of the metapopulation occurs when the migration levels are intermediate, a result that agrees well with previous studies on two-map CML systems.     

The effect of deoxycholate on yeast cell permeability

1. (1)
   Мембраны ненарушенных клеток дрожжей непроницаемы для солей янтарной кислоты и феррицианида, так что в их суспензии не наблюдается сукцинодегидрогеназы с феррикианидом как акцептором электронов.     
   
   

Phospholipase C activation by ethanol in rat hepatocytes is unaffected by chronic ethanol feeding.

The activation of phosphoinositide-specific phospholipase C by ethanol was compared in hepatocytes isolated from ethanol-fed rats and from pair-fed control animals. Ethanol (100-300 mM) caused a dose-dependent transient increase in cytosolic free Ca2+ levels in indo-1-loaded hepatocytes from both groups of animals. The rate of Ca2+ increase was similar in hepatocytes from control and ethanol-fed rats, but the decay of the Ca2+ increase was somewhat slower in the latter preparation. The ethanol-induced Ca2+ increase caused activation of glycogen phosphorylase, with 50% response at 50 mM-ethanol and a maximal response at 150-200 mM-ethanol, not significantly different in hepatocytes from control and ethanol-fed animals. Ins(1,4,5)P3 formation in response to ethanol (300 mM) or vasopressin (2 nM or 40 nM) was also similar in the two preparations. It is concluded that long-term ethanol feeding does not lead to an adaptive response with respect to the ethanol-induced phospholipase C activation in rat hepatocytes. The ability of ethanol in vitro to decrease membrane molecular order in liver plasma membranes from ethanol-fed and control rats was measured by e.s.r. Membranes from ethanol-fed animals had a significantly lower baseline order parameter compared with control preparations (0.313 and 0.327 respectively), indicative of decreased membrane molecular order. Addition of 100 mM-ethanol significantly decreased the order parameter in control preparations by 2.1%, but had no effect on the order parameter of plasma membranes from ethanol-fed rats, indicating that the plasma membranes had developed tolerance to ethanol, similar to other membranes in the liver. Thus the membrane structural changes associated with this membrane tolerance do not modify the ethanol-induced activation of phospholipase C. The transient activation of phospholipase C by ethanol in hepatocytes may play a role in maintaining an adaptive phenotype in rat liver.     

The effect of oral contraceptives on rat platelet membrane glycoproteins

Female rats were administered oral contraceptives and the levels of sialic acid on platelet membrane and granule glycoproteins were compared to controls using a sialic acid assay and a fluorescein-conjugated wheat germ agglutinin binding assay and also by measuring the binding of ¹²⁵I-labelled wheat germ agglutinin to glycoprotein bands from platelets separated by polyacrylamide electrophoresis. The contraceptive-treated rats showed increased levels of glycoprotein sialylation which may partly explain the altered physiological function of the platelets.     

Docosahexaenoic acid synthesis from alpha-linolenic acid by rat brain is unaffected by dietary n-3 PUFA deprivation

Rates of conversion of alpha-linolenic acid (alpha-LNA, 18:3n-3) to docosahexaenoic acid (DHA, 22:6n-3) by the mammalian brain and the brain's ability to upregulate these rates during dietary deprivation of n-3 polyunsaturated fatty acids (PUFAs) are unknown. To answer these questions, we measured conversion coefficients and rates in post-weaning rats fed an n-3 PUFA deficient (0.2% alpha-LNA of total fatty acids, no DHA) or adequate (4.6% alpha-LNA, no DHA) diet for 15 weeks. Unanesthetized rats in each group were infused intravenously with [1-(14)C]alpha-LNA, and their arterial plasma and microwaved brains collected at 5 minutes were analyzed. The deficient compared with adequate diet reduced brain DHA by 37% and increased brain arachidonic (20:4n-6) and docosapentaenoic (22:5n-6) acids. Only 1% of plasma [1-(14)C]alpha-LNA entering brain was converted to DHA with the adequate diet, and conversion coefficients of alpha-LNA to DHA were unchanged by the deficient diet. In summary, the brain's ability to synthesize DHA from alpha-LNA is very low and is not altered by n-3 PUFA deprivation. Because the liver's reported ability is much higher, and can be upregulated by the deficient diet, DHA converted by the liver from circulating alphaLNA is the source of the brain's DHA when DHA is not in the diet.     

The action of tropic adenohypophyseal hormones when administered intraventricularly on rat behavioral reactions

In experiments on rats the influence was studied of tropic hormones of adenohypophysis--somatotropin, thyrotropin and adrenocorticotropin (STH, ThTN and ACTH respectively) on the elaboration and extinction of conditioned reaction of active avoidance (CRAA) in Y-maze and behaviour in the open field under microinjections of hormones into the brain lateral ventricle (0.001-0.002 ME). It has been shown that all studied hormones improve animals learning at negative pain reinforcement; ThTH and ACTH retard it in contrast to STH, which accelerates the extinction of the elaborated CRAA; tropic hormones exert differentiated influence on rats behaviour in the open field. No distinct correlation was found between behavioural manifestation and the level of catecholamines in the rats hypothalamus.     

Gliding motility of algae is unaffected by cytochalasin B

The effect of the antibiotic cytochalasin B on gliding motility of Oscillatoria and Navicula was studied using time lapse microcinematography. Contrary to its effects on many other non-muscle motile systems, cytochalasin B did not significantly inhibit motility.     

The phosphorus status of northern hardwoods differs by species but is unaffected by nitrogen fertilization

Northern hardwood forests in the eastern US exhibit species-specific influences on nitrogen (N) cycling, suggesting that their phosphorus (P) cycling characteristics may also vary by species. These characteristics are increasingly important to understand in light of evidence suggesting that atmospheric N deposition has increased N availability in the region, potentially leading to phosphorus limitation. We examined how P characteristics differ among tree species and whether these characteristics respond to simulated N deposition (fertilization). We added NH₄NO₃ fertilizer (50 kg ha^?1 year^?1) to single-species plots of red oak (Quercus rubra L.), sugar maple (Acer saccharum Marsh.), eastern hemlock (Tsuga canadensis (L.) Carr.), American beech (Fagus grandifolia Ehrh.), and yellow birch (Betula alleghaniensis Britt.), in the Catskill Mountains, New York from 1997 to 2007. Species differences were observed in foliar, litter and root P concentrations, but all were unaffected by a cumulative N fertilization of 550 kg/ha. Similarly, measures of soil P availability and biotic P sufficiency differed by species but were unaffected by N fertilization. Results suggest species exhibit unique relationships to P as well as N cycles. We found little evidence that N fertilization leads to increased P limitation in these northern hardwood forests. However, species such as sugar maple and red oak may be sufficient in P, whereas beech and hemlock may be less sufficient and therefore potentially more sensitive to future N-stimulated P limitation.     

Selective inactivation of lactate dehydrogenase of rat tissues by sodium deoxycholate.

Soluble lactate dehydrogenase (EC 1.1.1.27) extracted from brain, skeletal and cardiac muscle and liver of rats, and purified isoenzymes LDH-1 and LDH-5, were incubated with sodium deoxycholate. Deoxycholate almost totally inactivated isoenzyme LDH-5 (A4), whereas it left isoenzyme LDH-1 (B4) unaffected. Tissue lactate dehydrogenase was inactivated to different degrees depending on the origin of the enzyme. Electrophoretic isoenzyme studies of tissue lactate dehydrogenase showed the loss of activity to be quantitatively related to the overall percentage of subunit A distributed among the homotetramer LDH-5 and the heterotetramers LDH-2, LDH-3 and LDH-4. It was concluded that subunit A of lactate dehydrogenase interacts selectively with deoxycholate, irrespective of its association with subunit B. Distinct changes in electrophoretic mobilities of deoxycholate-treated isoenzymes strongly indicated an indiscriminate binding of deoxycholate by all LDH isoenzymes, probably through hydrophobic interactions. The results suggest that the inactivation of the enzyme is non-competitive, but the basis of the selectivity of deoxycholate towards subunit A is not known at present.     

Oxidative enzyme activities in rat brain subcellular fractions. The effect of deoxycholate, freeze-thaw, and water

(1) The distributions of four oxidative enzymes were studied in crude brain fractions. (2) Freeze-thaw cycle treatment and frozen storage of homogenate fractions gave apparent enhancement of cytochrome oxidase and NADH cytochrome c reductase activities. (3) Deoxycholate released cytochrome oxidase and NADH cytochrome c reductase activities from low-speed precipitates. The NADH diaphorase was enhanced to a small degree while NADPH cytochrome c reductase was not affected by deoxycholate. (4) Distilled water coupled with a single homogenization released trapped soluble enzymes and mitochondria and gave nearly maximal cytochrome oxidase activity as judged by deoxycholate treatment. The total distilled water activity of NADH cytochrome c reductase was much less than that of deoxycholate-stimulated fractions. The activities of other enzymes were not markedly affected by distilled water although their distribution was changed.     

Th2 differentiation is unaffected by Jagged2 expression on dendritic cells

Expression of the Jagged Notch ligands by dendritic cells (DCs) has been suggested to play a role in instructing Th2 responses. Supporting this hypothesis, we found that Jagged2 but not Jagged1 expression, correlates with the ability of DCs to induce Th2 responses. Jagged2 expression is up-regulated in response to the helminth soluble Schistosoma mansoni egg Ag, which conditions DCs to induce Th2 responses, and is markedly down-regulated following exposure to TLR agonists that generally promote Th1 responses. Conversely, Jagged1 expression is markedly induced by TLR ligation. Despite these correlations, suppression of expression of Jagged2 using retrovirally delivered small interfering RNA failed to affect the ability of DCs to induce Th2 cell differentiation either in vitro or in vivo. Moreover, retrovirally induced expression of Jagged2 did not enhance the ability of DCs to induce Th2 cell responses. Our data indicate that Jagged2 expression by DCs is not sufficient or required for Th2 cell differentiation.     

The neutrophil migration induced by tumour necrosis factor alpha in mice is unaffected by glucocorticoids

Macrophages harvested from the peritoneal cavities of rats release a neutrophil chemotactic factor (MNCF) in response to stimulation with Gram-negative bacterial lipopolysaccharide (LPS). MNCF has been shown to be active in rats treated with dexamethasone, a glucocorticoid that usually inhibits the neutrophil migration induced in this species by interleukin (IL)-1, tumour necrosis factor alpha (TNFalpha), IL-8, C5a and leukotriene B(4) (LTB(4)). Here we report that macrophages harvested from peritoneal cavities of mice, and stimulated in vitro with LPS, also release a factor that induces neutrophil migration in dexamethasone-treated animals. This chemotactic activity was neutralized by the incubation of the LPS-stimulated macrophage supernatants with a purified polyclonal IgG anti-mouse TNFalpha. In addition, significant amounts of TNF were detected in the supernatants. The neutrophil migration induced by intraperitoneal administration of recombinant murine TNFalpha was also unaffected by pretreatment of the mice with dexamethasone. Moreover, neutrophil migration induced by intraperitoneal injection of LPS was completely blocked by pretreatment of the mice with a monoclonal antibody against murine TNFalpha. In conclusion, our results support the hypothesis that, in contrast to the role of TNF in rats (where it indirectly induces neutrophil migration), in mice, it may be an important mediator in the recruitment of neutrophils to inflammatory sites.     

Enhancement by metronidazole of the antitumor effect of cyclophosphane

In experiments on mouse tumors (sarcoma-180, Ehrlich ascites tumor and hemoblastosis La) the ability of metronidazole to enhance the antitumor chemotherapeutic action of cyclophosphane was investigated. It was shown that metronidazole (1000 mg/kg) injected 60 min before cyclophosphane administration can elicit an almost two-fold increase in its radioprotective efficiency. The chemosensitizing effect of metronidazole depends on the drug concentration and the tumor type.     

脱氧胆酸钠体外对新生隐球菌胞外磷脂酶的作用

目的比较观察脱氧胆酸钠体外对自然生长条件下的新生隐球菌新生变种(CZYD7)胞外磷脂酶的抑制作用。方法在37℃下,用蛋黄平板法培养并测量菌株产生沉淀圈的大小,用沉淀圈比值(PZ值)比较磷脂酶活力的变化。结果对照组与脱氧胆酸钠不同浓度组(依次为0.1%、0.2%、0.3%和0.4%)的PZ值分别为0.390±0.050,0.782±0.038,0.875±0.074,0.914±0.048,0.885±0.091;脱氧胆酸钠各浓度组与对照组比较均有显著差异(P<0.01)。脱氧胆酸钠不同浓度组间PZ值比较,0.1%和0.2%、0.1%和0.3%、0.1%和0.4%组间有显著差异(P<0.05)。结论脱氧胆酸钠体外对新生隐球菌新生变种CZYD7株胞外磷脂酶具有抑制作用。