Determination of citrulline in human plasma, red blood cells and urine by electron impact (EI) ionization gas chromatography-mass spectrometry

A method was developed by using gas chromatography-mass spectrometry in the electron impact ionization mode to quantify citrulline in plasma, red blood cells (RBC) and urine. For all three fluids, citrulline was extracted on ion exchange resins, before derivatization to its propyl-heptaflorobutyryl-ester. Assay precision (coefficient of variation, CV) was <5%, recovery% was >90% and the within- and between-day CV were <10% on 200 microL of plasma and RBC, and 400 microL of urine. The current method allows for the detection of 20 pmol of natural citrulline in aqueous standards, and small volumes (<100 microL) of biological fluids.     

Gène CFTR et infertilité masculine en 2001 Conseil génétique, diagnostic prénatal, diagnostic pré-implantatoire

As the vas deferens is also absent in the majority of CF (cystic fibrosis) males, it has been proposed that CBAVD (Congenital Bilateral Absence of Vas Deferens) males may present an incomplete or mild form of CF. Many studies using more extensive mutation analysis have confirmed the role of CFTR gene defects: 80% of CBAVD patients carry one or two mutations. Each patient with a diagnosis of CBAVD should also be examined for pulmonary and pancreatic signs, and sweat tests should be performed. In couples with CBAVD linked to CFTR mutations, the risk of having children with CF or infertility is increased if the female is also a carrier. The woman should be screened for the most frequent CFTR mutations according to her ethnic background. After screening for 80% of the mutations responsible for CF, the residual risk of being a carrier with negative screening is: Z=h(1?a)/(1?ah)=1/120 considering a carrier frequency of 1/25 in the general population. In the case of positive screening, antenatal diagnosis by chorionic villus sampling may be proposed. However, in some situations it is difficult to predict the phenotypic consequences for the child, particularly when a severe transmutation of a variable allele is identified. As these couples require medically-assisted reproduction techniques, pre-implantation genetic diagnosis appears to be more appropriate than antenatal diagnosis. Only embryos that inherit the non-mutated maternal CFTR allele are replaced in the uterus. Examination of childre born to couples with CBAVD is mandatory: immunoreactive trypsinogen assay at 3 days of age, sweat test at 3 months and clinical examination, especially looking for signs of CF. Identification of CFTR mutations in a CBAVD patient has important consequences for his family. Each sibling has a 50% risk of being a carrier and a 25% risk of inheriting the same genotype. The genetic counsellor must inform these siblings about the possible risk of having CF children if they carry CFTR mutations and if their partner is also a carrier.     

Signature Lipids and Stable Carbon Isotope Analyses of Octopus Spring Hyperthermophilic Communities Compared with Those of Aquificales Representatives

The molecular and isotopic compositions of lipid biomarkers of cultured Aquificales genera have been used to study the community and trophic structure of the hyperthermophilic pink streamers and vent biofilm from Octopus Spring. Thermocrinis ruber, Thermocrinis sp. strain HI 11/12, Hydrogenobacter thermophilus TK-6, Aquifex pyrophilus, and Aquifex aeolicus all contained glycerol-ether phospholipids as well as acyl glycerides. The n-C_20:1 and cy-C₂₁ fatty acids dominated all of the Aquificales, while the alkyl glycerol ethers were mainly C_18:0. These Aquificales biomarkers were major constituents of the lipid extracts of two Octopus Spring samples, a biofilm associated with the siliceous vent walls, and the well-known pink streamer community (PSC). Both the biofilm and the PSC contained mono- and dialkyl glycerol ethers in which C₁₈ and C₂₀ alkyl groups were prevalent. Phospholipid fatty acids included both the Aquificales n-C_20:1 and cy-C₂₁, plus a series of iso-branched fatty acids (i-C_15:0 to i-C_21:0), indicating an additional bacterial component. Biomass and lipids from the PSC were depleted in ¹³C relative to source water CO₂ by 10.9 and 17.2‰, respectively. The C_20–21 fatty acids of the PSC were less depleted than the iso-branched fatty acids, 18.4 and 22.6‰, respectively. The biomass of T. ruber grown on CO₂ was depleted in ¹³C by only 3.3‰ relative to C source. In contrast, biomass was depleted by 19.7‰ when formate was the C source. Independent of carbon source, T. ruber lipids were heavier than biomass (+1.3‰). The depletion in the C_20–21 fatty acids from the PSC indicates that Thermocrinis biomass must be similarly depleted and too light to be explained by growth on CO₂. Accordingly, Thermocrinis in the PSC is likely to have utilized formate, presumably generated in the spring source region.     

Biogeochemistry of hypersaline microbial mats illustrates the dynamics of modern microbial ecosystems and the early evolution of the biosphere

Photosynthetic microbial mats are remarkably complete self-sustaining ecosystems at the millimeter scale, yet they have substantially affected environmental processes on a planetary scale. These mats may be direct descendents of the most ancient biological communities in which even oxygenic photosynthesis might have developed. Photosynthetic mats are excellent natural laboratories to help us to learn how microbial populations associate to control dynamic biogeochemical gradients.     

Comparative ecology of H2 cycling in sedimentary and phototrophic ecosystems

The simple biochemistry of H₂ is critical to a large number of microbial processes, affecting the interaction of organisms with each other and with the environment. The sensitivity of each of these processes to H₂ can be described collectively, through the quantitative language of thermodynamics. A necessary prerequisite is to understand the factors that, in turn, control H₂ partial pressures. These factors are assessed for two distinctly different ecosystems. In anoxic sediments from Cape Lookout Bight (North Carolina, USA), H₂ partial pressures are strictly maintained at low, steady-state levels by H₂-consuming organisms, in a fashion that can be quantitatively predicted by simple thermodynamic calculations. In phototrophic microbial mats from Baja California (Mexico), H₂ partial pressures are controlled by the activity of light-sensitive H₂-producing organisms, and consequently fluctuate over orders of magnitude on a daily basis. The differences in H₂ cycling can subsequently impact any of the H₂-sensitive microbial processes in these systems. In one example, methanogenesis in Cape Lookout Bight sediments is completely suppressed through the efficient consumption of H₂ by sulfate-reducing bacteria; in contrast, elevated levels of H₂ prevail in the producer-controlled phototrophic system, and methanogenesis occurs readily in the presence of 40 mM sulfate. This revised version was published online in August 2006 with corrections to the Cover Date.     

Reverse flux through cardiac NADP(+)-isocitrate dehydrogenase under normoxia and ischemia

Little is known about the role of mitochondrial NADP(+)-isocitrate dehydrogenase (NADP(+)-ICDH) in the heart, where this enzyme shows its highest expression and activity. We tested the hypothesis that in the heart, NADP(+)-ICDH operates in the reverse direction of the citric acid cycle (CAC) and thereby may contribute to the fine regulation of CAC activity (Sazanov and Jackson, FEBS Lett 344: 109-116, 1994). We documented a reverse flux through this enzyme in rat hearts perfused with the medium-chain fatty acid octanoate using [U-(13)C(5)]glutamate and mass isotopomer analysis of tissue citrate (Comte et al., J Biol Chem 272: 26117-26124, 1997). In this study, we assessed the significance of our previous finding by perfusing hearts with long-chain fatty acids and tested the effects of changes in O(2) supply. We showed that under all of these conditions citrate was enriched in an isotopomer containing five (13)C atoms. This isotopomer can only be explained by substrate flux through reversal of the NADP(+)-ICDH reaction, which is evaluated at 3-7% of flux through citrate synthase. Small variations in reversal fluxes induced by low-flow ischemia that mimicked hibernation occurred despite major changes in contractile function and O(2) consumption of the heart as well as citrate and succinate release rates and tissue levels. Our data show a reverse flux through NADP(+)-ICDH and support its hypothesized role in the fine regulation of CAC activity in the normoxic and O(2)-deprived heart.     

The importance of isometric maximum strength and peak rate-of-force development in sprint cycling

This study was designed to investigate the relationship of whole-body maximum strength to variables potentially associated with track sprint-cycling success. These variables included body composition, power measures, coach's rank, and sprint-cycling times. The study was carried out in 2 parts. The first part (n = 30) served as a pilot for the second part (n = 20). Subjects for both parts ranged from international-caliber sprint cyclists to local-level cyclists. Maximum strength was measured using an isometric midthigh pull (IPF). Explosive strength was measured as the peak rate-of-force development (IPRFD) from the isometric force-time curve. Peak power was estimated from countermovement (CMJPP) and static vertical jumps (SJPP) and measured by modified Wingate tests. Athletes were ranked by the U.S. national cycling coach (part 1). Sprint times (from a standing start) were measured using timing gates placed at 25, 82.5, 165, 247.5, and 330 m of an outdoor velodrome (part 2). Maximum strength (both absolute and body-mass corrected) and explosive strength were shown to be strongly correlated with jump and Wingate power. Additionally, maximum strength was strongly correlated with both coach's rank (parts 1 and 2) and sprint cycling times (part 2). The results suggest that larger, stronger sprint cyclists have an advantage in producing power and are generally faster sprint cyclists.