Hydrolysis of N'-benzoyl-d-arginine-p-nitroanilide by members of the Haloanaerobiaceae: additional evidence that Haloanaerobium praevalens is related to endospore-forming bacteria

Abstract Three out of the four described halophilic obligately anaerobic bacteria of the family Haloanaerobiaceae hydrolyze d -BAPA (N'-benzoyl- d -arginine-p-nitroanilide), while showing no or little l -BAPA hydrolyzing activity. This property was shown earlier to be characteristic only of non-halophilic Gram-positive endospore-forming bacteria of the genera Bacillus and Clostridium . These results suggest that Haloanaerobium praevalens , which has never been shown to produce endospores, but was shown to be related to the endosphere-forming representatives of the Haloanaerobiaceae on the basis of 16S rRNA nucleotide sequence data, shares other properties characteristics of the endospore-forming bacteria. Neither significant d -BAPA nor l -BAPA hydrolyzing activity was found in Sporohalobacter lortetii .     

Localization and Characterization of Nitric Oxide Synthase in the Rat Suprachiasmatic Nucleus: Evidence for a Nitrergic Plexus in the Biological Clock

Abstract: Behavioral and electrophysiological evidence indicates that the biological clock in the hypothalamic suprachiasmatic nuclei (SCN) can be reset at night through release of glutamate from the retinohypothalamic tract and subsequent activation of nitric oxide synthase (NOS). However, previous studies using NADPH-diaphorase staining or immunocytochemistry to localize NOS found either no or only a few positive cells in the SCN. By monitoring conversion of l -[³H]arginine to l -[³H]citrulline, this study demonstrates that extracts of SCN tissue exhibit NOS specific activity comparable to that of rat cerebellum. The enzymatic reaction requires the presence of NADPH and is Ca²⁺/calmodulin-dependent. To distinguish the neuronal isoform (nNOS; type I) from the endothelial isoform (type III), the enzyme activity was assayed over a range of pH values. The optimal pH for the reaction was 6.7, a characteristic value for nNOS. No difference in nNOS levels was seen between SCN collected in day versus night, either by western blot or by enzyme activity measurement. Confocal microscopy revealed for the first time a dense plexus of cell processes stained for nNOS. These data demonstrate that neuronal fibers within the rat SCN express abundant nNOS and that the level of the enzyme does not vary temporally. The distribution and quantity of nNOS support a prominent regulatory role for this nitrergic component in the SCN.