The fungal species Metarhizium pingshaense, M. anisopliae, M. robertsii, and M. brunneum, a monophyletic group informally referred to as the PARB species complex, are well known facultative entomopathogens, including many commercialized strains used for biological pest control. Accurate and expedient species identification of Metarhizium isolates represents an important first step when addressing ecological as well as application-related questions involving these fungi. To this end, a species-specific multiplexed polymerase chain reaction (PCR) assay was developed for identification and discrimination among Metarhizium PARB complex species, based on unique sequence signature differences within the nuclear ribosomal intergenic spacer (rIGS) and nuclear intergenic spacer regions MzFG546 and MzIGS2. Species-specificities of the four primer pairs were assessed following a three-step approach including: (1) in silico verification of sequence signatures by BLASTN searches against publically available genome and amplicon sequence data, (2) corroboration of assay specificity and robustness by performing test PCR amplifications against a taxonomically curated reference strain collection of 68 Metarhizium strains representing 12 species, and (3) testing against a field collection of 19 unknown Metarhizium isolates from soil of a Swiss meadow. The specificity of these four primer pairs provide an efficient means to detect and discriminate PARB species in studies targeting ecological aspects of indigenous isolates, as well as efficacy, persistence and potential non-target effects of applied biocontrol strains.

Many patients with sporadic Alzheimer's disease (AD) suffer from memory impairment, anxiety- and depression. The systemic utility of N-Methyl-d-Aspartate (NMDA) receptor antagonists has been shown to be potential therapeutic target for memory loss in AD. However, there is no evidence that shows whether NMDA receptor antagonists have the same effects when these blockers are directly used within the brain regions including hippocampus. It might be an urgent to further explore the therapeutic role of NMDA receptor antagonists in behavioral abnormalities such as anxiety and depression in AD. The aim of this study was to determine whether blockade of the hippocampal NMDA receptors could attenuate neurobehavioral abnormalities in rats with sporadic AD. Twelve days after AD induction by streptozotocin (STZ), animals received either vehicle or MK-801 (NMDA receptor antagonist) in the hippocampus for 10 days. Two or five days after the last MK-801 treatment, spatial memory, anxiety- and depression-related behaviors, and inflammatory cytokines (interleukin-(IL)-6, IL-1β and tumor necrosis factor (TNF)-α) were evaluated. Our findings indicated that STZ treatment significantly elevated hippocampal inflammation, impaired spatial memory, and increased anxiety- and depression-related symptoms in rats. Interestingly, the hippocampal NMDA receptor blockade improved these neurobehavioral phenotypes and decreased inflammatory cytokines in the hippocampus of STZ-treated rats. Hippocampal NMDA receptors might be involved in neurobehavioral abnormalities via inflammation in sporadic AD.

Bifidobacteria represent one of the first colonizers of human gut microbiota, providing to this ecosystem better health and nutrition. To maintain a mutualistic relationship, they have enzymes to degrade and use complex carbohydrates non-digestible by their hosts. To succeed in the densely populated gut environment, they evolved molecular strategies that remain poorly understood. Herein, we report a novel mechanism found in probiotic Bifidobacteria for the depolymerization of the ubiquitous 2-acetamido-2-deoxy-4-O-(β-d-mannopyranosyl)-d-glucopyranose (Man-β-1,4-GlcNAc), a disaccharide that composes the universal core of eukaryotic N-glycans. In contrast to Bacteroidetes, these Bifidobacteria have a specialist and strain-specific β-mannosidase that contains three distinctive structural elements conferring high selectivity for Man-β-1,4-GlcNAc: a lid that undergoes conformational changes upon substrate binding, a tryptophan residue swapped between the two dimeric subunits to accommodate the GlcNAc moiety, and a Rossmann fold subdomain strategically located near to the active site pocket. These key structural elements for Man-β-1,4-GlcNAc specificity are highly conserved in Bifidobacterium species adapted to the gut of a wide range of social animals, including bee, pig, rabbit, and human. Together, our findings uncover an unprecedented molecular strategy employed by Bifidobacteria to selectively uptake carbohydrates from N-glycans in social hosts.

Diabetic nephropathy is the commonly developed complication of vasculature in type 2 diabetic patients. Chronic hyperglycemia leads to nephropathy in diabetics because of the formation of excessive reactive oxygen species and advanced glycation end products which is reflected in the form of glomerulosclerosis, tubular atrophy and interstitial fibrosis. As per the various reports reduction in SIRT1 expression in kidney tissue is key factor in the development of nephropathy in diabetes because its reduction in tissue is linked with excessive formation of ROS. Formononetin is a polyphenolic compound reported for its effect on SIRT1 and ROS.

Electroconvulsive therapy has been used successfully in some individuals with posttraumatic stress disorder (PTSD) whose symptoms have not improved with other treatments. But there are only a few reports. Meanwhile, an array of new neuromodulation strategies, including repetitive transcranial magnetic stimulation, transcranial direct current stimulation, vagus nerve stimulation, trigeminal nerve stimulation, and deep brain stimulation have been developed and applied experimentally in the treatment of other psychiatric disorders. This article will review the clinical evidence and mechanistic basis for their use in PTSD.

Vitamin D plays an important role in maintaining skeletal development and bone homeostasis. Although vitamin D has been extensively researched, the direct effect of 1,25(OH)2D3 on osteoblasts is unclear. To explore the 1,25(OH)2D3 action on murine osteoblasts, we performed tandem mass tag experiments on MC3T3-E1 cells treated with and without 1,25(OH)2D3. Three up-regulated proteins (MRC2, WWTR1 and RASSF2) related to bone metabolism were confirmed in this study. 1,25(OH)2D3 up-regulated the expression of MRC2 through vitamin D receptor. MRC2 affects collagen metabolism in osteoblasts. Combined with bioinformatics and parallel reaction monitoring analysis, we inhibited the expression of MRC2 to explore the relationship between MRC2 and collagens. Then we found MRC2 down-regulated COL5A2 and up-regulated MMP13. This study provides a protein profile of 1,25(OH)2D3-treated murine osteoblasts, reveals a newly discovered signaling axis (1,25(OH)2D3/VDR/MRC2/COL5A2 and MMP13), and explains the effect of 1,25(OH)2D3 on bone metabolism from a new perspective.

C1q is an important immune gene that can mediate a variety of immune regulatory functions, and is involved in complement pathway activation. In the present study, a ghC1q gene from the razor clam Sinonovacula constricta was identified and named ScghC1q-1. The complete ScghC1q-1 gene is 692 bp in length, with an open reading frame (ORF) of 489 bp encoding a protein of 162 amino acids. ScghC1q-1 mRNA was widely expressed in various tissues, and transcript levels in the hemolymph were significantly up-regulated following Staphylococcus aureus or Vibrio anguillarum challenge. Recombinant ScghC1q-1 protein was found to agglutinate both Gram-positive and Gram-negative bacteria. These results indicate that ScghC1q-1 plays an essential role in the immune defense of S. constricta.

Plant cell proliferation is affected by microgravity during spaceflight, but involved molecular mechanisms, key for space agronomy goals, remain unclear. To investigate transcriptomic changes in cell cycle phases caused by simulated microgravity, an Arabidopsis immobilized synchronous suspension culture was incubated in a Random Positioning Machine. After simulation, a transcriptomic analysis was performed with two subpopulations of cells (G2/M and G1 phases enriched) and an asynchronous culture sample. Differential expression was found at cell proliferation, energy/redox and stress responses, plus unknown biological processes gene ontology groups. Overall expression inhibition was a common response to simulated microgravity, but differences peak at the G2/M phase and stress response components change dramatically from G2/M to the G1 subpopulation suggesting a differential adaptation response to simulated microgravity through the cell cycle. Cell cycle adaptation using both known stress mechanisms and unknown function genes may cope with reduced gravity as an evolutionary novel environment.

Histone acetylation is an important post-translational modification in eukaryotes and is regulated by two antagonistic enzymes, namely histone acetyltransferase (HAT) and histone deacetylase (HDAC). However, little has been done on the HAT superfamily in Brachypodium distachyon (B. distachyon), a new model plant of Poaceae. In this study, eight HATs were identified from B. distachyon and classified into four major families. Subcellular localization analysis showed that a majority of BdHATs were predominantly localized in the nucleus. Syntenic and phylogenetic analysis indicated there may be two common ancestral CREB-binding protein (p300/CBP, HAC) genes prior to the separation of monocots and dicots. Expression analysis revealed that the potential roles of BdHATs in B. distachyon development and responses to four abiotic stresses. Protein-protein network analysis identified some potential interactive genes with BdHATs. Thus, our results will provide solid basis for further study the function of HAT genes in B. distachyon and other monocot plants.

During amoeboidal migration, cancer cells migrate in a protease-independent manner by squeezing through pre-existing gaps in the extracellular matrix (ECM). However, the extent to which cells alter their physical properties in order to sustain this mode of migration remains unclear. Here, we address this question by documenting biophysical changes in the properties of highly invasive MDA-MB-231 and HT-1080 cells upon inhibition of pericellular proteolysis. Remarkably, treatment with the broad spectrum MMP inhibitor GM6001 not only induces cell rounding and loss of actomyosin contractility, but also induces nuclear softening via increased phosphorylation of the nuclear membrane protein lamin A/C. Though nuclear softening is necessary for sustaining migration through sub-nuclear sized transwell pores, it is not sufficient. In addition, baseline levels of contractility mediating pore entry and peri-nuclear actin inside the pores mediating pore migration are also required. Taken together, our results suggest that protease-independent migration through sub-nuclear sized pre-existing tracks is enabled by deformation of a softened nucleus by contractility and the peri-nuclear actin network.

Epigenetic mechanisms regulate gene expression and may influence the pathogenesis of type 2 diabetes through the loss of insulin sensitivity. The aims of this study were to measure variation in DNA methylation at the type 2 diabetes locus KCNQ1 and assess its relationship with metabolic measures and with genotype.

High temperature is a common stress, which influences the growth and reproduction of plants. Maize is one of the most important crops all over the world. However, heat stress reduces significantly the yield and quality of maize. Therefore, it is important to illuminate molecular mechanism of maize response to heat stress. To estimate genes related to heat stress, we analyzed the transcriptome of maize in response to heat stress. In this study, six cDNA libraries were constructed form total RNA isolated from leaves of maize. A total of 35,209,446 and 35,205,472 clean reads were generated from CK (Control condition) and HTP (Heat stress condition) treatments, respectively. The results showed that 1857 DEGs were identified in maize after heat stress (1029 up-regulated and 828 down-regulated). KEGG pathway enrichment analysis for DEGs indicated that protein processing in endoplasmic reticulum pathways play a central role in maize response to heat stress. In addition, in the present study, 167 putative TFs were identified, which belong to various TF families (e.g., MYB, AP2-EREBP, b-ZIP, bHLH, NAC and WRKY), and may be associated with heat stress response of maize. This research may contribute to understand the molecular mechanism of maize inbred line B73 response to heat stress, which is beneficial for developing maize cultivars to improve yield and quality.

Ion uptake mechanisms are diverse in fish species, certainly linked to duplication events that have led to the presence of a multitude of paralogous genes. In fish, Na+ uptake involves several ion transporters expressed in different ionocyte subtypes. In the European sea bass Dicentrarchus labrax, several key transporters potentially involved in Na+ uptake have been investigated in seawater (SW) and following a 2 weeks freshwater (FW) acclimation. Using gel electrophoresis, we have shown that the Na+/H+-exchanger 3 (nhe3, slc9a3) is expressed in gills and kidney at both salinities. Quantitative realtime PCR analysis showed a significantly higher nhe3 expression in fresh water (FW) compared to SW. Its apical localization in a subset of gill ionocytes in freshwater-acclimated fish supports the role of NHE3 in Na+ uptake. Interestingly, NHE3-immunopositive cells also express basolateral Na+/K+/2Cl- cotransporter 1 (NKCC1) and are mainly localized in gill lamella. Among the three nhe2 (slc9a2) paralogs, only nhe2c shows differential branchial expression levels with higher mRNA levels in SW than in FW. The increased branchial expression of the ammonia transporter rhcg1 (Rhesus protein), nhe3 and cytoplasmic carbonic anhydrase (cac) in FW could indicate the presence of a functional coupling between ion transporters to form a Na+/NH4+ exchange complex. Acid-sensing ion channel 4 (asic4) seems not to be expressed in sea bass gills. Na+/Cl- cotransporter (ncc2a or ncc-like) is about three times more expressed in FW compared to SW suggesting coupled Na+ and Cl- uptake in a subset of gill ionocytes. Besides the main pump Na+/K+-ATPase, branchial NCC2a and NHE3 may be key players in ion uptake in sea bass following a long-term freshwater challenge.

In living organisms, the MicroRNAs act as an important role by controlling regulatory mechanisms, and likely manipulating the output of numerous protein-coding genes. Several computational databases, algorithms and tools have been developed to discover the miRNA target genes. But, the existing methods obtain poorer results in identification of miRNA target gene. Hence in this research work, integrated prediction scores is used to identify the microRNA target interactions and hybrid fuzzy algorithms are used to make final predictions. The proposed algorithms such as Fuzzy, Fuzzy + Support Vector Machine (SVM) and Fuzzy + SVM + Random Forest (RF) are used to conduct prediction by majority voting and it is compared with the existing techniques such as SVM, RF and Neural Network (NN) to demonstrate the performance of the proposed algorithms. Furthermore, the ranking features are estimated using the Arabidopsis thaliana microRNA sequence. From the experimental results, it is inferred that the proposed Fuzzy + SVM + RF algorithm performs superior than the existing ones.

Human plasma lipoproteins are known to contain various glycan structures whose composition and functional importance are starting to be recognized. We assessed N-glycosylation of human plasma HDL and LDL and the role of their glycomes in cellular cholesterol metabolism.

Viral aetiologies are the most common cause of central nervous system (CNS) infections. Approximately one-half of CNS infections remain of undetermined origin. High-throughput sequencing (HTS) brought new perspectives to CNS infection investigations, allowing investigation of viral aetiologies with an unbiased approach. HTS use is still limited to specific clinical situations.

Prasugrel as a second generation P2Y12 adenosine diphosphate receptor antagonist which in the cerebral aneurysms with Endovascular treatment have become more emphasized.

Insulin resistance and low-grade inflammation are pathophysiological mechanisms shared by type 2 diabetes and dementia. A cluster of biomarkers that could help diagnosing cognitive dysfunction prior to the installation of insulin resistance is desirable. This ELSA sub-study examined whether a cluster of selected inflammatory biomarkers was associated with worse cognitive scores in non-diabetic participants.

The molecular chaperon MRJ (DNAJB6) exhibits two splice isoforms that have different roles in human viral infection, but the regulatory mechanism of MRJ isoform expression is yet unclear. In this study, we show that reduction of the polyadenylation factor CstF64 was correlated with the increase of the MRJ large isoform (MRJ-L) in human macrophages and elucidate the mechanism underlying CstF64-modulated MRJ isoform expression. Moreover, we exploited an antisense strategy targeting MRJ-L for virus replication. A morpholino oligonucleotide complementary to the 5' splice site of MRJ intron 8 downregulated MRJ-L expression and suppressed the replication of not only HIV-1 but also respiratory syncytial virus (RSV). We demonstrated that downregulation of the MRJ-L level reduced HIV-1 replication as well as the subgenomic mRNA and viral production of RSV. The present findings that two human health-threatening viruses take advantage of MRJ-L for infection suggest MRJ-L as a potential target for broad-spectrum antiviral strategy.

BACKGROUND The prevalence and intensity of schistosomiasis infection in China has decreased markedly in recent years. Therefore, more accurate methods are critically needed to ensure further control of low-intensity schistosomiasis infection. For chronic schistosomiasis patients, the detection of schistosome eggs in colorectal mucosa tissues is commonly used. This work aimed to explore differences in sensitivity of the Schistosoma japonicum (S. japonicum) retrotransposon (SjR2) gene in colon tissue from S. japonicum infected hosts and to develop an ideal method for genetic diagnosis of low-intensity schistosomiasis. MATERIAL AND METHODS Serum and colon samples were collected from mice at different time points, either post-infection (PI) or post-treatment (PT). Colorectal biopsy specimens from outpatients with schistosomiasis were collected. All samples from mice and patients, including serum as well as colon tissue containing eggs and tissue containing no eggs, were examined using the polymerase chain reaction technique. RESULTS The results showed that the SjR2 gene could be detected in all colon tissue containing at least one egg, except for when the egg was completely degraded. The positive rate of gene detection in serum was low. The results from egg-free colon tissue from around the eggs were more consistent with the actual parasitism in vivo. CONCLUSIONS The results indicate that detection of the gene in colon tissue located within a 0.5 cm distance from the eggs would be a practical and ideal method for genetic diagnosis of schistosomiasis. After the colorectal biopsy, this method can be a sensitive assisted examination to the clinical diagnosis of low-intensity schistosomiasis infection.