Inhibitory effects of microalgae on the activation of hyaluronidase

The inhibitory effects of seven microalgae, Nostoc flagelliforme, Spirulina platensis, Porphyridium purpureum, Rhodosorus marinus,Chlorella pyrenoidosa, Dunaliella salina and Pleurochrysiscarterae on the activation of hyaluronidase were evaluated. Theinhibitory effect of the ethanol-insoluble fraction of each water extract frommicroalgae was stronger than that of the ethanol-soluble fraction. The50% inhibitory concentration (IC₅₀) of the ethanol-insolublefraction of S. platensis, P. purpureum, R. marinus, C.pyrenoidosa, D. salina and P. carterae was 0.15, 0.18, 0.26,0.94, 0.15 and 0.41 mg mL^-1, respectively. The IC₅₀ ofN .flagelliforme was not calculated, because there was no detectableinhibitory effect of this alga. The IC₅₀ of disodium cromoglycate(DSCG) used as the anti-allergic medicine was 0.14 mg mL^-1. The IC₅₀ of S. platensis, P. purpureum and D. salinawere almost the same as that of DSCG. This suggests that theethanol-insoluble fraction of S. platensis, P. purpureum and D. salina might be an anti-allergic substance. The ethanol-insoluble fractionof S. platensis and D. salina was ultrafiltered through a membranehaving a molecular exclusion limit of 20 kDa. The IC₅₀ of theresidue was stronger than that of the filtrate. These results suggest that theanti-allergic substance(s) of these microalgae may be polysaccharides.     

Activation of Rho in the injured axons following spinal cord injury

Axons of the adult central nervous system have very limited ability to regenerate after injury. This inability may be, at least partly, attributable to myelin-derived proteins, such as myelin-associated glycoprotein, Nogo and oligodendrocyte myelin glycoprotein. Recent evidence suggests that these proteins inhibit neurite outgrowth by activation of Rho through the neurotrophin receptor p75^NTR/Nogo receptor complex. Despite rapidly growing knowledge on these signals at the molecular level, it remained to be determined whether Rho is activated after injury to the central nervous system. To assess this question, we establish a new method to visualize endogenous Rho activity in situ. After treatment of cerebellar granular neurons with the Nogo peptide in vitro, Rho is spatially activated and colocalizes with p75^NTR. Following spinal cord injury in vivo, massive activation of Rho is observed in the injured neurites. Spatial regulation of Rho activity may be necessary for axonal regulation by the inhibitory cues.     

Identification of oxidative stress-regulated genes in rat aortic smooth muscle cells by suppression subtractive hybridization

Transfer RNA modification improves the rate of aa-tRNA selection at the A-site and the fitness in the P-site and thereby prevents frameshifting according to a new model how frameshifting occurs [Qian et al. (1998) Mol. Cell 1, 471-482]. Evidence that the presence of various modified nucleosides in tRNA also influences central metabolism, thiamine metabolism, and bacterial virulence is reviewed.     

Effect of DNA sequence divergence on homologous recombination as analyzed by a random-walk model

A point connecting a pair of homologous regions of DNA duplexes moves along the homology in a reaction intermediate of the homologous recombination. Formulating this movement as a random walk, we were previously successful at explaining the dependence of the recombination frequency on the homology length. Recently, the dependence of the recombination frequency on the DNA sequence divergence in the homologous region was investigated experimentally; if the methyl-directed mismatch repair (MMR) system is active, the logarithm of the recombination frequency decreases very rapidly with an increase of the divergence in a low-divergence regime. Beyond this regime, the logarithm decreases slowly and linearly with the divergence. This "very rapid drop-off" is not observed when the MMR system is defective. In this article, we show that our random-walk model can explain these data in a straightforward way. When a connecting point encounters a diverged base pair, it is assumed to be destroyed with a probability that depends on the level of MMR activity.     

Mechanisms of the protective effect of L-alanine to D-galactosamine-induced hepatocellular injury: comparative studies of L-alanine and pyruvate

The addition of L-alanine reduced lactate dehydrogenase leakage from primary cultured rat hepatocytes treated with galactosamine (D-gal), while D-alanine and other amino acids did not. However, the mechanisms have not yet been entirely clarified. In this study, we used various inhibitors of metabolism, i.e., aminooxyacetate, oligomycin, and quinolinic acid, to examine the relation between this protective effect and the metabolism of L-alanine. Quinolinic acid (10 mM) did not affect the hepatoprotective effect of L-alanine, while oligomycin (0.1 mug/ml) and aminooxyacetate (1 mM) eliminated the hepatoprotective effect of L-alanine. L-Alanine also increased the albumin secretion by cultured hepatocytes treated with D-gal, while pyruvate had little effect. It was revealed that the intracellular content of pyruvate did not increase as a result of addition of L-alanine. These results are consistent with the hypothesis that L-alanine metabolism is important for hepatoprotection, but pyruvate cannot be used as a substitute for L-alanine.     

Leucine promotes glucose uptake in skeletal muscles of rats

Soleus muscles isolated from normal rats were incubated to evaluate whether or not leucine promotes glucose uptake under insulin-free conditions, using a labeled 2-deoxyglucose uptake assay. Glucose uptake was promoted by 2mM leucine. A metabolite of leucine, alpha-ketoisocaproic acid (alpha-KIC), also exhibited a similar stimulatory effect, although this was not as potent as leucine. Stimulation of glucose uptake by leucine was completely canceled by pre-treatment with either 10 microM LY294002, a specific inhibitor of phosphatidylinositol 3-kinase (PI3-kinase), or 6 microM GF109203X, a specific inhibitor of protein kinase C (PKC). No significant change was observed by pre-treatment with 1 microM rapamycin, a specific inhibitor of mammalian target of rapamycin (mTOR). These results suggest that leucine stimulates glucose transport in skeletal muscle via PI3-kinase and PKC pathways independently of the mammalian target of mTOR. They also suggest that leucine stimulates glucose transport by an insulin-independent mechanism.     

Asymmetric random walk in a reaction intermediate of homologous recombination

At an intermediate step of the homologous recombination between two double-stranded DNA molecules, a point (often called Holliday structure) connecting two strands coming from two recombining partners migrates along the homologous region. Assuming random walk of a connecting point, we previously explained the dependence of recombination frequency on the homology length observed in vivo. In this model, the random walk was assumed to be symmetric in that the forward transition rate equals the backward one. According to observations in vitro, however, catalysed migration appears unidirectional. Taking into account possible asymmetry, we thus reformulate our random walk model to reexamine the observations in vivo. We also derive some theoretical results to analyse dynamic processes observed in vitro.     

Angiogenic activity of osteopontin-derived peptide SVVYGLR

Angiogenesis plays an important role in various pathological conditions as well as some physiological processes. Although a number of soluble angiogenic factors have been reported, extracellular matrix also has crucial effect on angiogenesis through interaction with endothelial cells. Since recent reports showed osteopontin had some angiogenic activity, the effect of the SVVYGLR peptide, novel binding motif in osteopontin molecule, on angiogenesis was examined in this study. Synthetic peptide SVVYGLR did not have proliferative effect on endothelial cells but adhesion and migration activity to endothelial cells. Furthermore, SVVYGLR had as potent activity for tube formation in three-dimensional collagen gel as vascular endothelial growth factor which is known to be the strongest angiogenic factor. Electron microscopical analysis showed a number of microvilli on the endothelial luminar surface and tight junction formation in the luminar intercellular border between endothelial cells, indicating SVVYGLR induced cell porarity and differentiation of endothelial cells. This small peptide might be expected to stimulate angiogenesis to improve some ischemic conditions in the future because of some advantages due to smaller molecular weight.