Sorption Behavior of Cesium in Two Greek Soils: Effects of Cs Initial Concentration,Clay Mineralogy,and Particle-size Fraction

The characteristics of Cs sorption behavior in two soils (soil 1 and soil 2) with nearly the same clay content and exhangeable K concentration, but with different clay mineralogy, were studied by the quantification of the distribution coefficient (k_d). It was observed that as the initial Cs concentration increased from 4 to 50 mg L^?1, the k_d values decreased in both soils, suggesting a progressive saturation of Cs available sorption sites. However, the presence of expansible 2:1 phyllosilicates minerals in the clay fraction of soil 2 maintained a high Cs sorption ability for this soil, even at high Cs concentrations. The experimental data were also fitted to the Freundlich isotherm and the results showed that parameters of the Freundlich equation could be used to estimate the degree of Cs sorption and the nature of the available sorption sites. For the studied soils, the k_f and the k_d values followed a similar trend and the n Freundlich constant values provided a reliable indicator for the soils’ clay mineralogy. The removal of the sand fraction enhanced Cs sorption in both soils and the absence of sorbed Cs ions on the quartz minerals, as observed by the SEM analysis, additionally supported the effect of particle-size fraction on Cs sorption.     

Positive genetic interactors of HMG2 identify a new set of genetic perturbations for improving sesquiterpene production in Saccharomyces cerevisiae

Background

Production of correctly disulfide bonded proteins to high yields remains a challenge. Recombinant protein expression in Escherichia coli is the popular choice, especially within the research community. While there is an ever growing demand for new expression strains, few strains are dedicated to post-translational modifications, such as disulfide bond formation. Thus, new protein expression strains must be engineered and the parameters involved in producing disulfide bonded proteins must be understood.

Results

We have engineered a new E. coli protein expression strain named SHuffle, dedicated to producing correctly disulfide bonded active proteins to high yields within its cytoplasm. This strain is based on the trxB gor suppressor strain SMG96 where its cytoplasmic reductive pathways have been diminished, allowing for the formation of disulfide bonds in the cytoplasm. We have further engineered a major improvement by integrating into its chromosome a signal sequenceless disulfide bond isomerase, DsbC. We probed the redox state of DsbC in the oxidizing cytoplasm and evaluated its role in assisting the formation of correctly folded multi-disulfide bonded proteins. We optimized protein expression conditions, varying temperature, induction conditions, strain background and the co-expression of various helper proteins. We found that temperature has the biggest impact on improving yields and that the E. coli B strain background of this strain was superior to the K12 version. We also discovered that auto-expression of substrate target proteins using this strain resulted in higher yields of active pure protein. Finally, we found that co-expression of mutant thioredoxins and PDI homologs improved yields of various substrate proteins.

Conclusions

This work is the first extensive characterization of the trxB gor suppressor strain. The results presented should help researchers design the appropriate protein expression conditions using SHuffle strains.     

Olympic weightlifting training causes different knee muscle-coactivation adaptations compared with traditional weight training

The purpose of this study was to compare the effects of an Olympic weightlifting (OL) and traditional weight (TW) training program on muscle coactivation around the knee joint during vertical jump tests. Twenty-six men were assigned randomly to 3 groups: the OL (n = 9), the TW (n = 9), and Control (C) groups (n = 8). The experimental groups trained 3 d · wk(-1) for 8 weeks. Electromyographic (EMG) activity from the rectus femoris and biceps femoris, sagittal kinematics, vertical stiffness, maximum height, and power were collected during the squat jump, countermovement jump (CMJ), and drop jump (DJ), before and after training. Knee muscle coactivation index (CI) was calculated for different phases of each jump by dividing the antagonist EMG activity by the agonist. Analysis of variance showed that the CI recorded during the preactivation and eccentric phases of all the jumps increased in both training groups. The OL group showed a higher stiffness and jump height adaptation than the TW group did (p < 0.05). Further, the OL showed a decrease or maintenance of the CI recorded during the propulsion phase of the CMJ and DJs, which is in contrast to the increase in the CI observed after TW training (p < 0.05). The results indicated that the altered muscle activation patterns about the knee, coupled with changes of leg stiffness, differ between the 2 programs. The OL program improves jump performance via a constant CI, whereas the TW training caused an increased CI, probably to enhance joint stability.     

Soluble CD40 ligand as a biomarker for storage-related preanalytic variations of human serum

There are currently no appropriate and sensitive biomarkers available to assess preanalytic variations in human biological fluids stored in biobanks. We identified soluble CD40 ligand (sCD40L) as the first ubiquitous biomarker to show an on-off response in serum exposed to moderate or elevated room temperature conditions. We used immunoenzyme assays to monitor the sCD40L response after 12 h storage at 37 degrees C or 48 h at 20 degrees C. Our findings show that prolonged storage of serum samples at elevated room temperature can be determined by the absence of detectable sCD40L.     

Plasminogen activator inhibitor type 1 inhibits smooth muscle cell proliferation in pulmonary arterial hypertension

RATIONALE: Pulmonary arterial smooth muscle cells (PASMCs) in the medial layer of the vessel wall are responsible for vessel homeostasis, but also for pathologic vascular remodelling in diseases, such as idiopathic pulmonary arterial hypertension (IPAH). Vascular remodelling in IPAH results in vessel stiffness, occlusion, and increased vascular resistance, but its underlying mechanisms remain to be fully elucidated. In this study, we investigated the expression and function of plasminogen activator inhibitor (PAI)-1, an inhibitor of the plasminogen activator system and target gene of the transforming growth factor (TGF)-beta1 signalling cascade, in PASMC in IPAH. METHODS AND RESULTS: RNA and protein analysis from lung tissues of donors and patients with IPAH (n=7 each) revealed a significant downregulation of PAI-1 in IPAH lungs. Immunohistochemical analysis localised PAI-1 to the bronchial and alveolar epithelium, as well as to vascular and airway smooth muscle cells. PAI-1 was also downregulated in primary PASMC derived from IPAH lungs as compared with donor-derived PASMC. In order to elucidate PAI-1 function, primary PASMC were stimulated with active recombinant (r)PAI-1, or transfected with PAI-1-specific siRNA. Stimulation with rPAI-1 led to decreased PASMC proliferation and adhesion to vitronectin, and increased PASMC migration. In contrast, PAI-1 knock-down with siRNA increased PASMC proliferation and decreased PASMC migration. CONCLUSIONS: PAI-1 is significantly downregulated in PASMC in IPAH, on the mRNA and protein level. PAI-1 negatively regulates PASMC proliferation, while it increases PASMC migration. Thus, its loss in IPAH may therefore contribute to pathologic vascular remodelling in IPAH.     

Metaproteome plasticity sheds light on the ecology of the rumen microbiome and its connection to host traits

The arsenal of genes that microbes express reflect the way in which they sense their environment. We have previously reported that the rumen microbiome composition and its coding capacity are different in animals having distinct feed efficiency states, even when fed an identical diet. Here, we reveal that many microbial populations belonging to the bacteria and archaea domains show divergent proteome production in function of the feed efficiency state. Thus, proteomic data serve as a strong indicator of host feed efficiency state phenotype, overpowering predictions based on genomic and taxonomic information. We highlight protein production of specific phylogenies associated with each of the feed efficiency states. We also find remarkable plasticity of the proteome both in the individual population and at the community level, driven by niche partitioning and competition. These mechanisms result in protein production patterns that exhibit functional redundancy and checkerboard distribution that are tightly linked to the host feed efficiency phenotype. By linking microbial protein production and the ecological mechanisms that act within the microbiome feed efficiency states, our present work reveals a layer of complexity that bears immense importance to the current global challenges of food security and sustainability.Subject terms: Microbial ecology, Microbiome     

Ala45Thr polymorphism of the NEUROD1 gene and diabetes susceptibility: a meta-analysis

A meta-analysis assessed whether the Ala45Thr polymorphism of the neurogenic differentiation 1 (NEUROD1) gene is associated with increased risk of diabetes mellitus type 1 (T1D) or type 2 (T2D). Fourteen case-control studies were analyzed, including genotype data on 3,057 patients with diabetes (T1D n=1,213, T2D n=1,844) and 2,446 controls. Overall and race-specific summary odds ratios (ORs) were obtained with fixed and random effects models. The Thr allele did not significantly increase the overall risk for T1D (OR 1.27 [0.94–1.71], P=0.12) or T2D (OR 1.07 [0.90–1.28], P=0.46). The Thr allele conferred increased susceptibility in subjects of Asian racial descent to T1D (OR 1.88 [1.10–3.21], P=0.020), but not to T2D (OR 1.08 [0.74–1.56], P=0.70). There was no association in subjects of European descent (OR 0.97 [0.76–1.23], P=0.80 for T1D; OR 1.03 [0.88–1.21], P=0.68 for T2D). Larger studies seemed to show more conservative estimates for the association with T1D (P=0.083). The Ala45Thr polymorphism of the NEUROD1 gene has no effect on susceptibility to T2D. It may however be a risk factor for susceptibility to T1D, in particular for subjects of Asian descent, although bias cannot be totally excluded.     

Recent advances in protein engineering and biotechnological applications of glutathione transferases

Glutathione transferases (GSTs, EC 2.5.1.18) are a widespread family of enzymes that play a central role in the detoxification, metabolism, and transport or sequestration of endogenous or xenobiotic compounds. During the last two decades, delineation of the important structural and catalytic features of GSTs has laid the groundwork for engineering GSTs, involving both rational and random approaches, aiming to create new variants with new or altered properties. These approaches have expanded the usefulness of native GSTs, not only for understanding the fundamentals of molecular detoxification mechanisms, but also for the development medical, analytical, environmental, and agricultural applications. This review article attempts to summarize successful examples and current developments on GST engineering, highlighting in parallel the recent knowledge gained on their phylogenetic relationships, structural/catalytic features, and biotechnological applications.