Description: Brucella abortus and Yersinia enterocolitica serotype O:9 serologically cross-react in the immune response with the host; therefore, our aim was to compare the immune responses to these two pathogens. We selected typical B. abortus and Y. enterocolitica O:9 strains to study the cytokine immune response and the histopathological changes in livers and spleens of BALB/c mice. The data showed the cytokine responses to the two strains of pathogens were different, where the average levels of granulocyte-macrophage colony-stimulating factor (GM-CSF), gamma interferon (IFN-), interleukin-12 (IL-12), and tumor necrosis factor alpha (TNF-α) were higher with B. abortus infections than with Y. enterocolitica O:9 infections, especially for IFN-, while the IL-10 level was lower and the levels of IL-1β, IL-4, IL-5, and IL-6 were similar. The histopathological effects in the livers and spleens of the BALB/c mice with B. abortus and Y. enterocolitica O:9 infections were similar; however, the pathological changes in the liver were greater with B. abortus infections, while damage in the spleen was greater with Y. enterocolitica O:9 infections. These observations show that different cytokine responses and histopathological changes occur with B. abortus and Y. enterocolitica O:9 infections.

Description: The etiology of uterine sarcomas: a pooled analysis of the epidemiology of endometrial cancer consortium British Journal of Cancer 108, 727 (19 February 2013). doi:10.1038/bjc.2013.2 Authors: A S Felix, L S Cook, M M Gaudet, T E Rohan, L J Schouten, V W Setiawan, L A Wise, K E Anderson, L Bernstein, I De Vivo, C M Friedenreich, S M Gapstur, R A Goldbohm, B Henderson, P L Horn-Ross, L Kolonel, J V Lacey, X Liang, J Lissowska, A Magliocco, M L McCullough, A B Miller, S H Olson, J R Palmer, Y Park, A V Patel, J Prescott, R Rastogi, K Robien, L Rosenberg, C Schairer, X Ou Shu, P A van den Brandt, R A Virkus, N Wentzensen, Y-B Xiang, W-H Xu, H P Yang & L A Brinton

Description: Bacterial operons are nature’s tool for regulating and coordinating multi-gene expression in prokaryotes. They are also a gene architecture commonly used in the biosynthesis of many pharmaceutically important compounds and industrially useful chemicals. Despite being an important eukaryotic production host, Saccharomyces cerevisiae has never had such gene architecture. Here, we report the development of a system to assemble and regulate a multi-gene pathway in S. cerevisiae . Full pathways can be constructed using pre-made parts from a plasmid toolbox. Subsequently, through the use of a yeast strain containing a stably integrated gene switch, the assembled pathway can be regulated using a readily available and inexpensive compound—estradiol—with extremely high sensitivity (10 nM). To demonstrate the use of the system, we assembled the five-gene zeaxanthin biosynthetic pathway in a single step and showed the ligand-dependent coordinated expression of all five genes as well as the tightly regulated production of zeaxanthin. Compared with a previously reported constitutive zeaxanthin pathway, our inducible pathway was shown to have 50-fold higher production level.

Description: Background— Circulating branched-chain amino acids and aromatic amino acids were recently related to insulin resistance and diabetes mellitus in prospective cohorts. We tested the effects of a genetic determinant of branched-chain amino acid/aromatic amino acid ratio on changes in body weight and insulin resistance in a 2-year diet intervention trial. Methods and Results— We genotyped the branched-chain amino acid/aromatic amino acid ratio—associated variant rs1440581 near the PPM1K gene in 734 overweight or obese adults who were assigned to 1 of 4 diets varying in macronutrient content. At 6 months, dietary fat significantly modified genetic effects on changes in weight, fasting insulin, and homeostasis model assessment of insulin resistance (HOMA-IR) after adjustment for the confounders (all P for interaction ≤0.006). Further adjustment for weight change did not appreciably change the interactions for fasting insulin and HOMA-IR. In the high-fat diet group, the C allele was related to less weight loss and smaller decreases in serum insulin and HOMA-IR (all P ≤ 0.02 in an additive pattern), whereas an opposite genotype effect on changes in insulin and HOMA-IR was observed in the low-fat diet group ( P =0.02 and P =0.04, respectively). At 2 years, the gene-diet interactions remained significant for weight loss ( P =0.008) but became null for changes in serum insulin and HOMA-IR resulting from weight regain. Conclusions— Individuals carrying the C allele of the branched-chain amino acid/aromatic amino acid ratio—associated variant rs1440581 may benefit less in weight loss and improvement of insulin sensitivity than those without this allele when undertaking an energy-restricted high-fat diet. Clinical Trial Registration— URL: http://www.clinicaltrials.gov . Unique identifier: NCT00072995.

Description: Based on an accurate atmospheric radiative transfer scheme, a parametrization of instantaneous global horizontal irradiance (GHI) at the Earth's surface has been developed. The scheme is named SUNFLUX and this article describes the development of the scheme for clear-sky conditions. The work dealing with clouds has been published in a separate article. Unlike traditional methods, this study applies the band model idea used in radiative transfer theory to the development of the surface radiation scheme and, importantly, includes absorption and scattering in the parametrization. Thus the scheme is more accurate compared with those using simple empirical approaches and may be applied to any site without being tuned for local conditions. The parametrization of aerosol transmittance and albedo developed by Kokhanovsky et al. is adopted to account for the effects of aerosols. All variables used in the scheme are available in climate models or from satellite observations. Therefore, the parametrization can be used to determine the GHI at the surface under clear-sky conditions The scheme is evaluated using observations obtained from three US Atmospheric Radiation Measurement (ARM) stations and three stations on the Tibetan Plateau, and the results demonstrate that the scheme is accurate. The relative mean bias difference is less than 4.3% and the relative root-mean-squared difference is less than 0.09%. Copyright © 2013 Royal Meteorological Society

Description: TNFR-associated factor (TRAF)6 is an essential ubiquitin E3 ligase in immune responses, but its function in adaptive immunity is not well understood. In this study, we show that TRAF6 is recruited to the peripheral ring of the T cell immunological synapse in Jurkat T cells or human primary CD4 + T cells conjugated with staphylococcal enterotoxin E–pulsed B cells. This recruitment depends on TRAF6 interacting with linker for activation of T cells (LAT) via its TRAF domain. Although LAT was indispensable for TCR/CD28-induced TRAF6 ubiquitination and its ligase activity, RNA interference–induced TRAF6 knockdown in T cells decreased TCR/CD28-induced LAT ubiquitination, tyrosine phosphorylation, and association with tyrosine kinase ZAP70. Overexpression of TRAF6 or its catalytically inactive form C70A promoted and decreased, respectively, LAT tyrosine phosphorylation upon stimulation. Moreover, LAT was ubiquitinated at Lys 88 by TRAF6 via K63-linked chain. In addition, TRAF6 was required for and synergized with LAT to promote the TCR/CD28-induced activation of NFAT. These results reveal a novel function and mechanism of TRAF6 action in the TCR–LAT signaling pathway distinct from its role in TCR-induced NF-B activation, indicating that LAT also plays an adapter role in TCR/CD28-induced activation of TRAF6.

Description: Earthward-propagating dipolarization fronts (DFs) are often found to be associated with magnetic reconnection and bursty bulk flows (BBFs) in the magnetotail. Recent THEMIS (Time History of Events and Macroscale Interactions During Substorms) probe observations have shown a DF propagating over 10 RE from the mid-tail region to the near-Earth tail region, and THEMIS All-Sky Imager data show a north-south auroral form and intensification of westward auroral zone currents. In this study, we examine THEMIS in situ observations of DFs in the magnetotail and simultaneous observations of the proton aurora from ground-based CANOPUS (the Canadian Auroral Network for the OPEN Program Unified Study) Meridian Scanning Photometers (MSPs). We find that earthward-moving DFs are often associated with intensification of proton aurora when the THEMIS probes are conjugate to the meridian of the MSP. The proton auroral intensifications are transient and in some cases detached from the background proton precipitation. Just before the DFs, the ion distribution is anisotropic in the field-aligned direction (mostly earthward) and the ion energy increases. These observations suggest that plasma sheet protons can be reflected and energized by earthward-moving DFs as they propagate through the magnetotail. We postulate that this population of ions is the source of the proton auroral intensification observed on the ground. This conjecture is tested using our global MHD simulation results, where the proton precipitation is calculated with the field-line curvature (FLC) model. The MHD simulation results show that proton precipitation enhancement can be caused by compression of plasma by approaching DFs/BBFs, which is consistent with ion reflection at DFs. Thus, using the conjugate observations from THEMIS spacecraft and MSP in this study, we are able to directly link the magnetotail dynamics, i.e., dipolarization fronts, with ground auroral activities. However, understanding of DF-associated ion energization requires detailed test-particle simulations with an analytical magnetotail model, such as those in our companion paper.

Description: Human slow skeletal troponin T (HSSTnT) shares a high degree of homology with cardiac TnT (CTnT). Although the presence of HSSTnT has not been confirmed in the heart at the protein level, detectable levels of HSSTnT mRNA have been found. Whether HSSTnT isoforms are expressed transiently remains unknown. Because transient re-expression of HSSTnT may be a potential mechanism of regulating function, we explored the effect of HSSTnT on the regulation of cardiac muscle. At least three HSSTnT isoforms have been found to exist in slow skeletal muscle: HSSTnT1 (+exons 5 and 12), HSSTnT2 (+exon 5, −exon 12), and HSSTnT3 (−exons 5 and 12). Another isoform, HSSTnT hypothetical (Hyp) (−exon 5, +exon 12), has only been found at the mRNA level. Compared with HCTnT3 (adult isoform), Tn complexes containing HSSTnT1, -2, and -3 did not alter the actomyosin ATPase activation and inhibition in the presence and absence of Ca2+, respectively. HSSTnTHyp was not evaluated as it did not form a Tn complex under a variety of conditions. Porcine papillary skinned fibers displaced with HSSTnT1, -2, or -3 and reconstituted with human cardiac troponin I and troponin C (HCTnI·TnC) complex showed a decrease in the Ca2+ sensitivity of force development and an increase in maximal recovered force (HSSTnT1 and -3) compared with HCTnT3. In contrast, HSSTnTHyp showed an increase in the Ca2+ sensitivity of force development. This suggests that re- or overexpression of specific SSTnT isoforms might have therapeutic potential in the failing heart because they increase the maximal force of contraction. In addition, circular dichroism and proteolytic digestion experiments revealed structural differences between HSSTnT isoforms and HCTnT3 and that HSSTnT1 is more susceptible to calpain and trypsin proteolysis than the other HSSTnTs. Overall, HSSTnT isoforms despite being homologues of CTnT may display distinct functional properties in muscle regulation.

Description: Neuropeptide Y (NPY) is implicated in the regulation of blood pressure (BP), and NPY pathways in the hypothalamus are sensitive to dietary fat. We evaluated the potential effect of a functional variant rs16147 located in the NPY gene promoter region on the association between 2-year diet intervention and change in multiple BP measures in the randomized Preventing Overweight Using Novel Dietary Strategies Trial. The NPY rs16147 was genotyped in 723 obese adults who were randomly assigned to 1 of 4 diets differing in the target percentages of energy derived from fat, protein, and carbohydrate. The changes of 4 BP phenotypes, including systolic BP, diastolic BP, pulse pressure, and mean arterial pressure, during 2-year diet intervention were analyzed. In the total participants and participants with hypertension, we observed significant and consistent interactions between rs16147 genotype and dietary fat intake on changes in multiple BP phenotypes at 2 years (all P for interactions 〈0.05). The risk allele (C allele) was associated with a greater reduction of BP phenotypes in response to low-fat diet, whereas an opposite genetic effect was observed in response to high-fat diet. In addition, the C allele was related to greater changes in 4 BP phenotypes in hypertensive compared with nonhypertensive participants. Our data suggest that NPY rs16147 may modulate the association between dietary fat intake and changes in BP phenotypes, and the C allele exerts a long-term beneficial effect on lowering BP in response to low-fat diet in obese and hypertensive subjects.

Description: Japanese encephalitis virus (JEV) is an enveloped flavivirus with a single-stranded, positive-sense RNA genome encoding three structural and seven nonstructural proteins. To date, the role of JEV nonstructural protein 2A (NS2A) in the viral life cycle is largely unknown. The interferon (IFN)-induced double-stranded RNA (dsRNA)-activated protein kinase (PKR) phosphorylates the eukaryotic translation initiation factor 2α subunit (eIF2α) after sensing viral RNA and results in global translation arrest as an important host antiviral defense response. In this study, we found that JEV NS2A could antagonize PKR-mediated growth inhibition in a galactose-inducible PKR-expressing yeast system. In human cells, PKR activation, eIF2α phosphorylation, and the subsequent translational inhibition and cell death triggered by dsRNA and IFN-α were also repressed by JEV NS2A. Moreover, among the four eIF2α kinases, NS2A specifically blocked the eIF2α phosphorylation mediated by PKR and attenuated the PKR-promoted cell death induced by the chemotherapeutic drug doxorubicin. A single point mutation of NS2A residue 33 from Thr to Ile (T33I) abolished the anti-PKR potential of JEV NS2A. The recombinant JEV mutant carrying the NS2A-T33I mutation showed reduced in vitro growth and in vivo virulence phenotypes. Thus, JEV NS2A has a novel function in blocking the host antiviral response of PKR during JEV infection.