A kinetic study of gamma-glutamyltransferase (GGT)-mediated S-nitrosoglutathione catabolism.

S-Nitrosoglutathione (GSNO) is a nitric oxide (NO) donor compound which has been postulated to be involved in transport of NO in vivo. It is known that c-glutamyl transpeptidase (GGT) is one of the enzymes involved in the enzyme-mediated decomposition of GSNO, but no kinetics studies of the reaction GSNO-GGT are reported in literature. In this study we directly investigated the kinetics of GGT with respect to GSNO as a substrate and glycyl- glycine (GG) as acceptor co-substrate by spectrophotometry at 334 nm. GGT hydrolyses the c-glutamyl moiety of GSNO to give S-nitroso-cysteinylglycine (CGNO) and c-glutamyl-GG. However, as both the substrate GSNO and the first product CGNO absorb at 334 nm, we optimized an ancillary reaction coupled to the enzymatic reaction, based on the copper-mediated decomposition of CGNO yielding oxidized cysteinyl-glycine and NO. The ancillary reaction allowed us to study directly the GSNO/GGT kinetics by following the decrease of the characteristic absorbance of nitrosothiols at 334 nm. A Km of GGT for GSNO of 0.398 ± 31 mM was thus found, comparable with Km values reported for other c-glutamyl substrates of GGT

T-786→C polymorphism of the endothelial nitric oxide synthase gene is associated with insulin resistance in patients with ischemic or non ischemic cardiomyopathy

Background: Insulin resistance (IR) and endothelial dysfunction are frequently associated in cardiac disease. The T-786→C variant in the promoter region of the endothelial nitric oxide synthase (eNOS) gene has been associated with IR in both non-diabetic and diabetic subjects. Aim of the study was to assess the reciprocal relationships between T-786→C eNOS polymorphism and IR in ischemic and non-ischemic cardiomyopathy.Method: A group of 132 patients (108 males, median age 65 years) with global left ventricular (LV) dysfunction secondary to ischemic or non-ischemic heart disease was enrolled. Genotyping of T-786→C eNOS gene promoter, fasting glucose, insulin, and insulin resistance (defined as HOMA-IR index > 2.5) were determined in all patients.Results: Genotyping analysis yielded 37 patients homozygous for the T allele (TT), 70 heterozygotes (TC) and 25 homozygous for C (CC). Patients with CC genotype had significantly higher systemic arterial pressure, blood glucose, plasma insulin and HOMA index levels than TT. At multivariate logistic analysis, the history of hypertension and the genotype were the only predictors of IR. In particular, CC genotype increased the risk of IR (CI% 1.4-15.0, p < 0.01) 4.5-fold. The only parameter independently associated with the extent of LV dysfunction and the presence of heart failure (HF) was the HOMA index (2.4 CI% 1.1-5.6, p < 0.04).Conclusions: T-786→C eNOS polymorphism was the major independent determinant of IR in a population of patients with ischemic and non-ischemic cardiomyopathy. The results suggest that a condition of primitive eNOS lower expression can predispose to an impairment of glucose homeostasis, which in turn is able to affect the severity of heart disease. © 2012 Vecoli et al.; licensee BioMed Central Ltd

Impaired myocardial metabolic reserve and substrate selection flexibility during stress in patients with idiopathic dilated cardiomyopathy.

Under resting conditions, the failing heart shifts fuel use toward greater glucose and lower free fatty acid (FFA) oxidation. We hypothesized that chronic metabolic abnormalities in patients with dilated cardiomyopathy (DCM) are associated with the absence of the normal increase in myocardial glucose uptake and maintenance of cardiac mechanical efficiency in response to pacing stress. In 10 DCM patients and 6 control subjects, we measured coronary flow by intravascular ultrasonometry and sampled arterial and coronary sinus blood. Myocardial metabolism was determined at baseline, during atrial pacing at 130 beats/min, and at 15 min of recovery by infusion of [(3)H]oleate and [(13)C]lactate and measurement of transmyocardial arteriovenous differences of oxygen and metabolites. At baseline, DCM patients showed depressed coronary flow, reduced uptake and oxidation of FFA, and preferential utilization of carbohydrates. During pacing, glucose uptake increased by 106% in control subjects but did not change from baseline in DCM patients. Lactate release increased by 122% in DCM patients but not in control subjects. Cardiac mechanical efficiency in DCM patients was not different compared with control subjects at baseline but was 34% lower during stress. Fatty acid uptake and oxidation did not change with pacing in either group. Our results show that in DCM there is preferential utilization of carbohydrates, which is associated with reduced flow and oxygen consumption at rest and an impaired ability to increase glucose uptake during stress. These metabolic abnormalities might contribute to progressive cardiac deterioration and represent a target for therapeutic strategies aimed at modulating cardiac substrate utilization

The climatic significance of Late Ordovician-early Silurian black

The Ordovician-Silurian transition (455-430 Ma) is characterized by repeated climatic perturbations, concomitant with major changes in the global oceanic redox state best exemplified by the periodic deposition of black shales. The relationship between the climatic evolution and the oceanic redox cycles, however, remains largely debated. Here, using an ocean-atmosphere general circulation model accounting for ocean biogeochemistry (MITgcm), we investigate the mechanisms responsible for the burial of organic carbon immediately before, during and right after the latest Ordovician Hirnantian (445-444 Ma) glacial peak. Our results are compared with recent sedimentological and geochemical data. We show that the late Katian time slice (445 Ma), typified by the deposition of black shales at tropical latitudes, represents an unperturbed oceanic state, with regional organic carbon burial driven by the surface primary productivity. During the Hirnantian, our experiments predict a global oxygenation event, in agreement with the disappearance of the black shales in the sedimentary record. This suggests that deep-water burial of organic matter may not be a tenable triggering factor for the positive carbon excursion reported at that time. Our simulations indicate that the perturbation of the ocean circulation induced by the release of freshwater, in the context of the post-Hirnantian deglaciation, does not sustain over sufficiently long geological periods to cause the Rhuddanian (444 Ma) oceanic anoxic event. Input of nutrients to the ocean, through increased continental weathering and the leaching of newly-exposed glaciogenic sediments, may instead constitute the dominant control on the spread of anoxia in the early Silurian

New sedimentological and biostratigraphic data in the Kwahu Group (Meso- to Neo-Proterozoic), southern margin of the Volta Basin, Ghana: Stratigraphic constraints and implications on regional lithostratigraphic correlations

Since several decades, geologists disagree about the lithostratigraphic subdivision of the Volta Basin sedimentary infilling. Correlations at the scale of the Volta Basin were up to now largely limited by the discontinuity of outcrops along the basin margin and by the scarcity of stratigraphic constraints within the sedimentary succession. Even if a subdivision of the Volta Basin infilling into three main groups (Bombouaka, Oti, and Obosum Groups) is now generally accepted, there is no agreement for lithostratigraphic subdivision at the formation scale. The Bombouaka Group, which represents first deposits of the Volta Basin sedimentary infilling, is particularly concerned. During mapping project in Ghana, authors proceeded to a sedimentological study of the Kwahu Group deposits on the Kwahu Plateau area. Some samples collected during field mapping were analyzed on a palynological point of view and revealed several layers rich in palynomorphs (acritarchs). Siliciclastic sediments composing the Kwahu Group are distinguished in 12 main facies associations, which are interpreted to be representative of both marine and continental environments. Sedimentological study of these facies associations leads to identify a cyclic vertical evolution of depositional settings at different scales, which are interpreted as sequences. The Kwahu Group is thus composed of three distinct macrosequences (about 300-500 m thick) interpreted as deltaic progradational sequences. Each macrosequence has been used to define formations for mapping, sequence stratigraphy key surfaces being considered as limits for cartographic units. The acritarch content of the analyzed samples allows rather coarse but nonetheless useful biostratigraphic constraints which, in association with data from sedimentological and sequential analysis, are used to improve correlations into the Bombouaka and Kwahu Groups at the scale of the Volta basin. The present data complemented with the few recently published studies on Voltaian deposits, lead to a new regional lithostratigraphic synthesis of the Bombouaka and Kwahu Groups. based on stratigraphic constrains and sequence stratigraphy. (C) 2011 Elsevier B.V. All rights reserved

REMARKS ON THE CAMBRIAN-ORDOVICIAN BOUNDARY IN THE ALGERIAN SAHARA (HASSI-RMEL AREA)

The question of the exact boundary between the Cambrian and Ordovician systems in the Algerian Sahara has never been clearly settled. The study of several early oilwells located between Laghouat and Ghardaia made possible an interpretation of the nature of the sequences and the environment of the lower Paleozoic, A thorough palynological study based on the Acritarchs Group was also performed on the NL 2 (Bordj Nili No 2) well. Satisfactory correlations with the standard scales used in Europe (Estonia) and North America (Eastern Canada) were found. The authors were able to reach valid conclusions concerning the limit between the two systems

Determination of S-nitrosoglutathione and other nitrosothiols by p-hydroxymercurybenzoate derivatization and reverse phase chromatography coupled with chemical vapor generation atomic fluorescence detection

NRC publication: Ye

Exogenous vs. endogenous gamma-glutamyltransferase activity: Implications for the specific determination of S-nitrosoglutathione in biological samples

The determination of S-nitrosoglutathione (GSNO) levels in biological fluids is controversial, partly due to the laborious sample handling and multiple pretreatment steps required by current techniques. GSNO decomposition can be effected by the enzyme gamma-glutamyltransferase (GGT), whose involvement in GSNO metabolism has been suggested. We have set up a novel analytical method for the selective determination and speciation of GSNO and its metabolite S-nitrosocysteinylglycine, based on liquid chromatography separation coupled to on-line enzymatic hydrolysis of GSNO by commercial GGT. In a post-column reaction coil, GGT allows the specific hydrolysis of the gamma-glutamyl moiety of GSNO, and the S-nitrosocysteinylglycine (GCNO) thus formed is decomposed by copper ions originating oxidized cysteinylglycine and nitric oxide (NO). NO immediately reacts with 4,5-diaminofluorescein (DAF-2) forming a triazole derivative, which is detected fluorimetrically. The limit of quantitation (LOQc) for GSNO and GCNO in plasma ultrafiltrate was 5 nM, with a precision (CV) of 1-6% within the 5-1500 nM dynamic linear range. The method was applied to evaluate the recovery of exogenous GSNO after addition of aliquots to human plasma samples presenting with different total GGT activities. By inhibiting GGT activity in a time dependent manner, it was thus observed that the recovery of GSNO is inversely correlated with plasmatic levels of endogenous GGT, which indicates the need for adequate inhibition of endogenous GGT activity for the reliable determination of endogenous GSNO

Chitinozoan biostratigraphy across the Katian (Late Ordovician) GICE event in the Borenshult-1 drillcore (Sweden)

The Late Ordovician is a period of major global environmental changes. Its understanding requires an integrated stratigraphical approach. Here, chitinozoan assemblages are described from the Borenshult-1 drillcore from Motala (Sweden) and related to stable carbon isotope event stratigraphy. Fifty-six rock samples, corresponding to a stratigraphic interval ranging from the upper Sandbian to Llandovery, were dissolved and analysed for chitinozoans. This stratigraphic interval covers the Sandbian to Llandovery and includes, in ascending order, the Freberga, Slandrom, Fjacka, Jonstorp, Loka and Motala formations. This study resulted in identification of two biozones the Spinachitina cervicomis Biozone (lower part of the Freberga Formation), as well as its Angochitina multiplex Subzone, and the Fungochitina spinifera Biozone (upper part of the Freberga Formation). The spinifera Biozone is characterized by the presence of Lagenochitina baltica, whereas the cervicomis Biozone is defined by the presence of its index species. Despite a low recovery of chitinozoans from Borenshult-1, we suggest that the Guttenberg Isotopic Carbon Excursion (GICE), as observed in the Borenshult-1 drillcore, correlates with the S. cervicomis Biozone. This fits well with chitinozoan and carbon isotope data from the Fjacka section in central Sweden (Dalarna, Siljan region) and from the Smedsby Gard drillcore (Ostergotland). The A. multiplex Subzone which is recognised in the Borenshult-1 and nearby Smedsby Gard drillcores, predates the GICE at both localities. (C) 2013 Elsevier B.V. All rights reserved