Regression of hypoxic hypertension-induced changes in the elastic laminae of rat pulmonary arteries

Liu, S. Q. Regression of hypoxic hypertension-inducedchanges in the elastic laminae of rat pulmonary arteries.J. Appl. Physiol. 82(5):1677-1684, 1997.The elastic laminae of the pulmonary arteries(PAs) undergo a progressive structural change in hypoxic hypertension.This study focused on the reversibility of altered PA elastic laminaeof the rat due to hypoxic hypertension. The structure andcross-sectional area of the PA medial elastic laminae were examined byusing electron-microscopic and image-analytic approaches duringrecovery from 12 h and 10 days of hypoxic hypertension. At 12 h ofhypoxic hypertension, the elastic laminae, which appeared homogeneousin normal control animals, were reorganized into structures composed ofrandomly oriented filaments, with an increase in the cross-sectionalarea of 70%. At 10 days of hypoxic hypertension, the elastic laminaeappeared homogeneous in structure and normal in cross-sectional areadespite continuous exposure to hypoxia. During recovery from 12 h ofhypoxic hypertension, the medial elastic laminae regained theirhomogeneous structure and normal cross-sectional area afterday 2. During recovery from 10 days ofhypoxic hypertension, the medial elastic laminae changed from homogeneous to filamentous structures, with a progressively altered cross-sectional area that increased by 89% from recoveryday 0 to day10 and returned to the normal level onday 30. These changes were associatedwith alterations in the PA wall tensile stress. These results indicatedthat structural changes in the PA elastic laminae were reversible andthat the regression process depended on the duration of exposure tohypoxia, the state of the elastic laminae, and possibly the tensilestress level in the PA wall.

     

An increase in nitric oxide produced by rat peritoneal neutrophils is not involved in cell apoptosis

Polymorphonuclear neutrophils (PMN) obtained from carrageenin-stimulated peritoneal cavities of rats, but not blood PMN, spontaneously produced nitric oxide (NO) when incubated in vitro. Incubation of the cells with the NO synthase inhibitors, L-imino-ethyl-L-ornithine (L-NIO) or N(G)-monomethyl-L-arginine (L-NMMA), inhibited NO production. This inhibition could be reversed by L-arginine. Incubation of PMN with lipopolysaccharide (LPS) failed to enhance NO production. Pretreatment of the rats with dexamethasone (DEXA) prior to carrageenin injection or incubation of PMN with the glucocorticoid in vitro partially inhibited the spontaneous release of NO. On the other hand, when PMN obtained from DEXA pretreated rats were incubated in vitro with DEXA, NO synthase activity and hence NO generation were almost abolished. A similar inhibition was also observed following the addition of L-NIO or cycloheximide to cultures of carrageenin-elicited PMN. The NO production by PMN did not appear to be related to cell viability or apoptosis. Indeed, neither the blockade of NO generation by L-NIO nor the incubation of the neutrophils with a NO donor, S-nitroso-acetylpenicillamine (SNAP) modified the pattern of LDH release or DNA fragmentation. In summary, it appears that PMN migration triggers a continuous NO synthesis, and that NO produced by these cells is not related to their apoptosis.     

Calcium-activated thiol-proteinase activity in the fusion of rat erythrocytes induced by benzyl alcohol.

1. Rat erythrocytes were fused by incubation with benzyl alcohol and Ca2+. 2. Cell fusion was inhibited by EGTA, N-ethylmaleimide, tetrathionate, iodoacetamide, cystamine, Tos-Lys-CH2Cl, and to a lesser extent by Tos-Phe-CH2Cl. Phenylmethanesulphonyl fluoride, Tos-Arg-OMe and histamine did not inhibit cell fusion. 3. Gel electrophoresis of membrane proteins from "ghosts" of the erythrocytes treated with benzyl alcohol showed that a high-molecular-weight polymer was present: this was consistent with the entry into the cells of Ca2+ and the activation of a transglutaminase enzyme. 4. In the treated cells the proteins corresponding to bands 2 and 3 in human erythrocytes were decreased, and a polypeptide with a slightly greater mobility than band 3 was produced. 5. These changes were inhibited by EGTA, N-ethylmaleimide, tetrathionate, iodoacetamide, cystamine, and Tos-Lys-CH2Cl, but not by phenylmethanesulphonyl fluoride, Tos-Arg-OMe, or histamine. 6. The intramembraneous particles of the P-fracture face of cells treated with benzyl alcohol to induce fusion were decreased in number and were susceptible to cold-induced aggregation; both of these phenomena were markedly inhibited to EGTA, and partially inhibited by Tos-Lys-CH2Cl and N-ethylmaleimide. 7. These several observations indicate that a Ca2+-activated thiol-proteinase, which acts to degrade membrane proteins and to give freedom of lateral movement to intramembranous particles, may be essential feature of membrane fusion in this system. 8. It is suggested that this proteinase may act to degrade spectrin-binding proteins that attach band-3 protein to the erythrocyte cytoskeleton.     

An animal model for the measurement of acute tolerance to ethanol

An apparatus is described for the measurement of acute tolerance to ethanol in small animals. Male, Sprague-Dawley rats were trained on the apparatus to leap to a descending platform to avoid being shocked. After an i.p. injection of 2 g/kg ethanol, the rats were tested repeatedly on the apparatus, and the plasma ethanol concentration was measured after each trial. The results demonstrated that the jumping ability of the rats was significantly more impaired during the ascending portion of the plasma ethanol curve than during the descending portion of the curve. Furthermore, it was demonstrated that the improvement in jumping ability during the descending portion of the curve was not dependent on a lowered plasma ethanol concentration. In a second experiment, the possibility of practice effects was eliminated by measuring the jumping ability and plasma ethanol concentration in one group of rats on the ascending portion of the plasma ethanol curve and in another group on the descending portion of the curve. A significant improvement in jumping ability was again observed during the descending portion of the curve, even though the plasma ethanol concentrations of the two groups were comparable. The development of acute tolerance to ethanol was thus demonstrated in both experiments.     

Exogenous C1q reconstitutes a secondary deficiency of C5-deficient AKR mouse macrophages for FcR-dependent cellular cytotoxicity and phagocytosis

Studies originally designed to assess the putative role of endogenous C5 in macrophage activation for antibody-dependent cellular cytotoxicity (ADCC) yielded unanticipated results. Resident and inflammatory peritoneal macrophages from C5-deficient AKR mice were found to have significantly lower capacity for FcR-dependent ADCC activation and phagocytosis of IgG-opsonized SRBC targets than did C5-competent C3HeB/FeJ (C3H) mice. Reconstitution of the ADCC response of AKR macrophages was accomplished initially with C5-sufficient C3H mouse serum, which suggested that endogenous C5 may be required for ADCC activation. However, further investigation largely eliminated C5 involvement in that a heat-labile component of C5-deficient AKR serum was shown to be active in the reconstitution of ADCC activation of AKR macrophages. Macrophages from AKR mice were found to have significantly lower levels of C1q mRNA synthesis, endogenous C1q levels, and C1q secretion than did C3H mouse macrophages as determined by Northern blot, Western blot, and presynthetic radiolabeling analysis, respectively. The addition of purified exogenous C1q to IgG-opsonized SRBC targets fully reconstituted ADCC activation for AKR inflammatory peritoneal macrophages to levels of normally FcR-responsive C3H macrophages. Similarly, exogenous C1q augmented FcR-dependent phagocytosis of AKR macrophages but had no effect on macrophages from responsive C3H mice. Our results indicate that AKR mice have a deficiency for FcR-dependent cellular cytotoxicity and phagocytosis that is related to their low potential for C1q synthesis and secretion rather than to their established genetic deficiency for C5 synthesis. We tentatively conclude that endogenous C1q is required as an accessory molecule for macrophage FcR-dependent effector functions and that C5 is not a prerequisite for ADCC activation.