Biochemical aspects of nitric oxide synthase feedback regulation by nitric oxide

Nitric oxide (NO) is a small gas molecule derived from at least three isoforms of the enzyme termed nitric oxide synthase (NOS). More than 15 years ago, the question of feedback regulation of NOS activity and expression by its own product was raised. Since then, a number of trials have verified the existence of negative feedback loop both in vitro and in vivo. NO, whether released from exogenous donors or applied in authentic NO solution, is able to inhibit NOS activity and also intervenes in NOS expression processes by its effect on transcriptional nuclear factor NF-κB. The existence of negative feedback regulation of NOS may provide a powerful tool for experimental and clinical use, especially in inflammation, when massive NOS expression may be detrimental

The Role of Protein Crystallography in Defining the Mechanisms of Biogenesis and Catalysis in Copper Amine Oxidase

Copper amine oxidases (CAOs) are a ubiquitous group of enzymes that catalyze the conversion of primary amines to aldehydes coupled to the reduction of O2 to H2O2. These enzymes utilize a wide range of substrates from methylamine to polypeptides. Changes in CAO activity are correlated with a variety of human diseases, including diabetes mellitus, Alzheimer’s disease, and inflammatory disorders. CAOs contain a cofactor, 2,4,5-trihydroxyphenylalanine quinone (TPQ), that is required for catalytic activity and synthesized through the post-translational modification of a tyrosine residue within the CAO polypeptide. TPQ generation is a self-processing event only requiring the addition of oxygen and Cu(II) to the apoCAO. Thus, the CAO active site supports two very different reactions: TPQ synthesis, and the two electron oxidation of primary amines. Crystal structures are available from bacterial through to human sources, and have given insight into substrate preference, stereospecificity, and structural changes during biogenesis and catalysis. In particular both these processes have been studied in crystallo through the addition of native substrates. These latter studies enable intermediates during physiological turnover to be directly visualized, and demonstrate the power of this relatively recent development in protein crystallography

Evidence-based Kernels: Fundamental Units of Behavioral Influence

This paper describes evidence-based kernels, fundamental units of behavioral influence that appear to underlie effective prevention and treatment for children, adults, and families. A kernel is a behavior–influence procedure shown through experimental analysis to affect a specific behavior and that is indivisible in the sense that removing any of its components would render it inert. Existing evidence shows that a variety of kernels can influence behavior in context, and some evidence suggests that frequent use or sufficient use of some kernels may produce longer lasting behavioral shifts. The analysis of kernels could contribute to an empirically based theory of behavioral influence, augment existing prevention or treatment efforts, facilitate the dissemination of effective prevention and treatment practices, clarify the active ingredients in existing interventions, and contribute to efficiently developing interventions that are more effective. Kernels involve one or more of the following mechanisms of behavior influence: reinforcement, altering antecedents, changing verbal relational responding, or changing physiological states directly. The paper describes 52 of these kernels, and details practical, theoretical, and research implications, including calling for a national database of kernels that influence human behavior

Racial and Sex Differences in 24 Hour Urinary Hydration Markers among Male and Female Emerging Adults: A Pilot Study

The purpose of this study was to examine 24 h urinary hydration markers in non-Hispanic White (WH) and non-Hispanic Black (BL) males and females. Thirteen males (BL, n = 6; WH, n = 7) and nineteen females (BL, n = 16, WH, n = 3) (mean ± SD; age, 20 ± 4 y; height, 169.2 ± 12.2 cm; body mass, 71.3 ± 12.2 kg; body fat, 20.8 ± 9.7%) provided a 24 h urine sample across 7 (n = 13) or 3 (n = 19) consecutive days (148 d total) for assessment of urine volume (UVOL), urine osmolality (UOSM), urine specific gravity (USG), and urine color (UCOL). UVOL was significantly lower in BL (0.85 ± 0.43 L) compared to WH college students (2.03 ± 0.70 L) (p < 0.001). Measures of UOSM, USG, and UCOL, were significantly greater in BL (716 ± 263 mOsm∙kg−1, 1.020 ± 0.007, and 4.2 ± 1.4, respectively) compared to WH college students (473 ± 194 mOsm∙kg−1, 1.013 ± 0.006, 3.0 ± 1.2, and respectively) (p < 0.05). Differences in 24 h urinary hydration measures were not significantly different between males and females (p > 0.05) or between the interaction of sex and race/ethnicity (p > 0.05). Non-Hispanic Black men and women were inadequately hydrated compared to their non-Hispanic White counterparts. Our findings suggest that development of targeted strategies to improve habitual fluid intake and potentially overall health are needed

Influence of Nutrient Intake on 24 Hour Urinary Hydration Biomarkers Using a Clustering-Based Approach

Previous work focusing on understanding nutrient intake and its association with total body water homeostasis neglects to consider the collinearity of types of nutrients consumed and subsequent associations with hydration biomarkers. Therefore, the purpose of this study was to analyze consumption patterns of 23 a priori selected nutrients involved in osmotic homeostasis, as well as their association with 24 h urinary hydration markers among fifty African–American first-year college students through a repeated measures observation in a daily living setting. Through application of hierarchical clustering, we were able to identity four clusters of nutrients based on 24 h dietary recalls: (1) alcohol + pinitol, (2) water + calcium + magnesium + erythritol + inositol + sorbitol + xylitol, (3) total calories + total fat + total protein + potassium + sodium + zinc + phosphorous + arginine, and (4) total carbohydrates + total fiber + soluble fiber + insoluble fiber + mannitol + betaine. Furthermore, we found that consumption of nutrients in Cluster #2 was significantly predictive of urine osmolality (p = 0.004); no other clusters showed statistically significant associations with 24 h urinary hydration biomarkers. We conclude that there may be some nutrients that are commonly consumed concomitantly (at the day level), across a variety of settings and populations, and that a limited subset of the clustering of these nutrients may associate with body water status

Phenyl Ether- and Aniline-Containing 2-Aminoquinolines as Potent and Selective Inhibitors of Neuronal Nitric Oxide Synthase

Excess nitric oxide (NO) produced by neuronal nitric oxide synthase (nNOS) is implicated in neurodegenerative disorders. As a result, inhibition of nNOS and reduction of NO levels is desirable therapeutically, but many nNOS inhibitors are poorly bioavailable. Promising members of our previously reported 2-aminoquinoline class of nNOS inhibitors, although orally bioavailable and brain-penetrant, suffer from unfavorable off-target binding to other CNS receptors, and they resemble known promiscuous binders. Rearranged phenyl ether- and aniline-linked 2-aminoquinoline derivatives were therefore designed to a) disrupt the promiscuous binding pharmacophore and diminish off-target interactions, and b) preserve potency, isoform selectivity, and cell permeability. A series of these compounds was synthesized and tested against purified nNOS, endothelial NOS (eNOS), and inducible NOS (iNOS) enzymes. One compound, 20, displayed high potency, selectivity, and good human nNOS inhibition, and retained some permeability in a Caco-2 assay. Most promisingly, CNS receptor counterscreening revealed this rearranged scaffold significantly reduces off-target binding