Measurement of organic-35S and organic-S in lake sediments: Methodological considerations

Three protocols for the determination of inorganic and organic sulfur fractions were tested for their suitability to estimate total indigenous organic sulfur (S_org) and³⁵S_org formed from added³⁵SO₄ ^2– in sediments of chemically dilute lakes in the ELA. The protocols tested have all been reported in the literature. It was found that two protocols involving sequential analyses for S fractions following acid treatment gave estimates of both S_org and³⁵S_org up to 87% lower than a non-sequential protocol. The low estimates were largely due to hydrolysis and solubilization of solid phase S which was then removed in a rinsing step. The non-sequential protocol, in which total reduced inorganic sulfur and total sulfur were determined on separate aliquots, is recommended as the most reliable of the three. Individual analyses in this protocol were verified for these lake sediments using a variety of S standards.     

Application of rRNA-based probes for observing marine nanoplanktonic protists.

The use of small-subunit rRNA-based oligonucleotides as probes for detecting marine nanoplanktonic protists was examined with a ciliate (an Uronema sp.), a flagellate (a Cafeteria sp.), and mixed assemblages of protists from enrichment cultures and natural seawater samples. Flow cytometry and epifluorescence microscopy analyses demonstrated that hybridizations employing fluorescein-labeled, eukaryote-specific probes intensely stained logarithmically growing protists, whereas these same protist strains in late stationary growth were barely detectable. The fluorescence intensity due to probe binding was significantly enhanced by the use of probes end labeled with biotin, which were detected by fluorescein-labeled avidin. The degree of signal amplification ranged from two- to fivefold for cultured protists in both logarithmic and stationary growth phases. Mixed assemblages of heterotrophic protists from enrichment cultures were also intensely labeled by rRNA-targeted oligonucleotide probes by the biotin-avidin detection system. Protists in late stationary growth phase and natural assemblages of protists that were otherwise undetectable when hybridized with fluorescein-labeled probes were easily visualized by this approach. In the latter samples, hybridization with multiple, biotin-labeled probes was necessary for detection of naturally occurring marine protists by epifluorescence microscopy. The signal amplification obtained with the biotin-avidin system should increase the utility of rRNA-targeted probes for identifying protists and facilitate characterization of the population structure and distribution of protists in aquatic environments.     

Kinetic and metabolic effects of nitrogen, magnesium and sulphur restriction in Xanthomonas campestris batch cultures

By reducing the concentration of nitrogen (from 5.0 to 2.5 mmol 1^-1), batch cultures of Xanthomonas campestris induced the enzyme UDP-glucose dehydrogenase and stimulated the Entner-Doudoroff pathway enzyme glucose-6-P dehydrogenase. The surplus energy generation was directed to xanthan biosynthesis resulting in a 10% polysaccharide increase. The nitrogen restriction led to a higher consumption of nitrogen (93%) whereas glucose consumption did not surpass 75% utilization. Low concentrations of both magnesium and sulphur exerted a negative effect on xanthan formation. Both restrictions reduced the phosphomannose isomerase enzyme activity by 10-fold turning the mannose transference presumably into the rate-limiting step for xanthan biosynthesis. Conversely, the rate of synthesis of glucuronic acid residues did not affect the rate of xanthan biosynthesis. Polysaccharide synthesis in magnesium and sulphur cultures was negatively affected in comparison with cell formation as the cell volumetric production rate increased from 0.037 to 0.091 g 1^-1 h^-1 and the xanthan volumetric production rate dropped from 0.133 g 1^-1 h^-1 to the minimum obtained at 0.083 g 1^-1 h^-1. The efficiency of the carbon substrate conversion was also greatly changed.     

A new approach for extracting information from protein dynamics

Increased ability to predict protein structures is moving research focus towards understanding protein dynamics. A promising approach is to represent protein dynamics through networks and take advantage of well-developed methods from network science. Most studies build protein dynamics networks from correlation measures, an approach that only works under very specific conditions, instead of the more robust inverse approach. Thus, we apply the inverse approach to the dynamics of protein dihedral angles, a system of internal coordinates, to avoid structural alignment. Using the well-characterized adhesion protein, FimH, we show that our method identifies networks that are physically interpretable, robust, and relevant to the allosteric pathway sites. We further use our approach to detect dynamical differences, despite structural similarity, for Siglec-8 in the immune system, and the SARS-CoV-2 spike protein. Our study demonstrates that using the inverse approach to extract a network from protein dynamics yields important biophysical insights.     

Plasma concentrations of choline in man following choline chloride

Plasma choline levels were measured in patients being treated with choline chloride for movement disorders. Following single doses of 5 g given orally in aqueous solution, plasma concentrations rose to a peak within four hours and then rapidly declined. The degree of increase was variable both between and within patients. During chronic treatment, plasma choline concentrations tended to rise as the dose increased, although the relationship was not strong. The highest concentrations attained by patients were always at a dose of 16 or 20 g daily. Following chronic treatment, the disappearance of choline from plasma was rapid, with most patients reaching baseline by four days. Choline chloride is generally given in four divided doses, which seems reasonable in the early stages of treatment. Most therapeutic effect is seen when patients are treated with daily doses in the 12 to 20 g range, doses likely to produce substantial increases in plasma choline concentration. However, the relationship of plasma choline concentration to clinical efficacy may be tenuous. Following discontinuation of treatment, clinical improvement tends to persist long after plasma choline has returned to baseline concentrations.     

Lipid-drug interaction: a structural analysis of pindolol effects on model membranes.

The ternary system constituted by distearoylphosphatidylcholine, pindolol (a vasodilator drug) and water has been investigated by using X-ray diffraction and calorimetric techniques. The structural modifications induced by the drug have been determined and a possible interaction model has been derived. In particular, the pindolol content-temperature dependent phase diagram shows the occurrence of two new phases: the first is an interdigitated gel, and the second is a lamellar structure presenting an unusual mixed disordered-ordered conformation of the hydrocarbon chains (L alpha beta). The comparative analysis of electron density profiles relative to the L alpha beta phase, reveals significant modifications in the paraffinic region of the lipid layer. In agreement with thermodynamic results, the structural data suggest that the drug induces a stiffening and a tightening of the hydrocarbon chains. Moreover, the hydrophilic properties of the membrane (particularly in P beta, and L alpha beta phases) present an evident dependence with the drug concentration.     

Diagrammatic scale for severity evaluation of the Eremanthus erythropappus–Puccinia velata pathosystem

Candeia (Eremanthus erythropappus (DC.) MacLeish), a native forest species from South America, has garnered commercial interest due to its production of essential oil that contains alpha-bisabolol. This compound is widely used in the pharmaceutical and cosmetics industry, with approximately 80% of Brazilian production being exported. Since candeia rust (Puccinia velata) has only been reported in Brazil, little is known about its epidemiology and control. There is no methodology to quantify rust severity in candeia, justifying the elaboration and validation of a diagrammatic scale containing eight levels of disease severity based on leaf area coverage (0.25%, 0.5%, 1%, 2%, 4%, 8%, 16% and 32%). In a natural sampling of disease in the field, 95% of the leaves showed severity below 16%, with the remaining 5% showing severities between 16% and 32% of leaf area. Validation of the proposed diagrammatic scale was performed by assessing the results from 10 inexperienced evaluators, performing evaluations of three leaves with different severity levels. The evaluations were performed at 7-day intervals; in the first instance, severity values were assigned without the diagrammatic scale, and for the second attempt, the scale proposed in this study was used. The accuracy and precision of the severity estimates produced by each evaluator compared to the real severity was analysed by linear regression and by Lin's statistics. The reproducibility of the estimates was evaluated by analysing the coefficient of determination of linear regressions by pairs of evaluators. The scale provided adequate levels of accuracy, precision, repeatability and reproducibility, indicating the proposed scale was a suitable method for quantifying the severity of candeia rust.     

S-adenosylmethionine induces mitochondrial dysfunction,permeability transition pore opening and redox imbalance in subcellular preparations of rat liver

Journal of Bioenergetics and Biomembranes - S-adenosylmethionine (AdoMet) predominantly accumulates in tissues and biological fluids of patients affected by liver dysmethylating diseases,...