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The article presents a case study of a 4-year old boy with an asymptomatic mass at the back of right thigh. The morphology and immunophenotype of the tumor corresponded to the characterized group of soft tissue spindle cell lesions defined by a relatively uniform cytomorphology, perivascular hyalinization, and S100 and CD34 coexpression. The article adds benefits of parallel sequencing as the types and number of gene fusions these tumors can render single-gene assay insensitive.

The expression of the myogenic regulatory factors (MRFs), Myf5, MyoD, myogenin ( Mgn) and MRF4 have been analysed during the development of chicken embryo somites and limbs. In somites, Myf5 is expressed first in somites and paraxial mesoderm at HH stage 9 followed by MyoD at HH stage 12, and Mgn and MRF4 at HH stage 14. In older somites, Myf5 and MyoD are also expressed in the ventrally extending myotome prior to Mgn and MRF4 expression. In limb muscles a similar temporal sequence is observed with Myf5 expression detected first in forelimbs at HH stage 22, MyoD at HH stage 23, Mgn at HH stage 24 and MRF4 at HH stage 30. This report describes the precise time of onset of expression of each MRF in somites and limbs during chicken embryo development, and provides a detailed comparative timeline of MRF expression in different embryonic muscle groups. [ABSTRACT FROM AUTHOR]

Background and Purpose: Icariin may be the active ingredient in Herba Epimedii, a Chinese herb commonly used for treatment of osteoporosis. The present study aims to delineate the mechanism(s) by which icariin prevents bone loss after ovariectomy (OVX) in vivo and stimulates osteoblastic functions in vitro.Experimental Approach: Ovariectomized or sham-operated C57BL/6 mice were treated with vehicle, 17beta-oestradiol or icariin for 6 weeks. Total and trabecular bome mineral density (BMD) as well as polar stress-strain index of distal femur were measured by peripheral computed tomography. The mRNA expressions of OPG and RANKL in tibia were studied by RT-PCR. Interactions between the oestrogen receptor (ER) antagonist ICI182,780 and icariin were studied in UMR 106 cells. The functional transactivation of ERalpha and ERbeta as well as ERalpha phosphorylation by icariin were also assessed.Key Results: Icariin suppressed the loss of bone mass and strength in distal femur and increased the mRNA expression ratio of OPG/RANKL in tibia, following OVX. Icariin increased ER-dependent cell proliferation, alkaline phosphatase (ALP) activity, gene expression of OPG and the OPG/RANKL ratio in UMR 106 cells. Icariin did not activate ERE-luciferase activity in UMR 106 cells, via the ERalpha or the ERbeta-mediated pathway, but it did increase ERalpha phosphorylation at Ser118.Conclusions and Implications: Our results indicate that icariin exerts anabolic effects in bone possibly by activating ER in a ligand-independent manner. Its ability to prevent OVX-induced bone loss without inducing uterotrophic effects supports its use as an alternative regimen for management of postmenopausal osteoporosis. [ABSTRACT FROM AUTHOR]

The modification of synaptic and neural connections in adults, including the formation and removal of synapses, depends on activity‐dependent synaptic and structural plasticity. MicroRNAs (miRNAs) play crucial roles in regulating these changes by targeting specific genes and regulating their expression. The fact that somatic and dendritic activity in neurons often occurs asynchronously highlights the need for spatial and dynamic regulation of protein synthesis in specific milieu and cellular loci. MicroRNAs, which can show distinct patterns of enrichment, help to establish the localized distribution of plasticity‐related proteins. The recent study using atomic force microscopy (AFM)‐based nanoscale imaging reveals that the abundance of miRNA(miR)‐134 is inversely correlated with the functional activity of dendritic spine structures. However, the miRNAs that are selectively upregulated in potentiated synapses, and which can thereby support prospective changes in synaptic efficacy, remain largely unknown. Using AFM force imaging, significant increases in miR‐132 in the dendritic regions abutting functionally‐active spines is discovered. This study provides evidence for miR‐132 as a novel positive miRNA regulator residing in dendritic shafts, and also suggests that activity‐dependent miRNAs localized in distinct sub‐compartments of neurons play bi‐directional roles in controlling synaptic transmission and synaptic plasticity. [ABSTRACT FROM AUTHOR]

Selective herbicides are used to control undesirable vegetation or weeds in fields without harming crops. Herbicide use for weed management can directly impact the densities of insect pests in agricultural communities as a result of insect mortality during and immediately after application. In insects, the glutathione S‐transferase (GST) enzyme is involved in both the detoxification process and the defense of cellular membranes against oxidative damage. In this study, two TmGSTs (TmGST‐iso1 and TmGST‐iso2) were identified and characterized to elucidate the GST family in Tenebrio molitor. Among the developmental stages of T. molitor, eggs had the highest expression levels of TmGST‐iso1. TmGST‐iso2 expression was highest in the pre‐pupal stage. TmGST‐iso1 expression was high in the guts of early and late larvae, whereas TmGST‐iso2 expression was not observed in early larvae. Adults, both male and female, had the highest levels of TmGST‐iso1 mRNA expression in the gut and reproductive organs, and the highest levels of TmGST‐iso2 expression in the reproductive organs. Quantitative polymerase chain reaction was used to investigate the impact of treatment with butachlor on the mRNA expression of TmGST‐iso1 and TmGST‐iso2 in larvae. TmGST‐iso1 expression increased in the butachlor‐treated group after 3 and 24 h, whereas TmGST‐iso2 expression peaked at 24 h after treatment. This study provides vital information about the detoxifying activities of T. molitor. [ABSTRACT FROM AUTHOR]

Agricultural intensification has led to significant increases in production, but the overuse of pesticides and associated hazards pose threats to biodiversity and ecological functions. Catalase (CAT), a key antioxidant enzyme, plays a crucial role in alleviating oxidative stress by directly interacting with toxins. In this study, we identified three CAT isoforms in Tenebrio molitor (TmCAT‐iso1, TmCAT‐iso2 and TmCAT‐iso3). These CATs possess a CAT domain, tetramer interface sites and a heme‐binding pocket. We examined the expression of Tm catalases across all developmental stages and in specific tissues using quantitative real time polymerase chain reaction (qRT‐PCR) experiments. Our findings demonstrate that TmCAT‐iso1 and TmCAT‐iso3 exhibit peak expression in young and late larval stages, respectively, whereas TmCAT‐iso2 shows peak expression during the egg and pre‐pupal stages. Tissue distribution analysis revealed the high expression of TmCAT‐iso1 and TmCAT‐iso2 in larval hemocytes, whereas TmCAT‐iso3 is predominantly expressed in larval Malpighian tubules. Furthermore, injection with chlorantraniliprole significantly elevated mRNA expression levels of TmCAT‐iso1, TmCAT‐iso2, and TmCAT‐iso3 in larval groups, compared with control groups. Our study highlights the distinct developmental stages and tissues where TmCATs are expressed. We also elucidated the effects of pesticide application on the expression of each TmCAT, revealing the physiological characteristics of CATs in response to these pesticides, which are dose‐ and time‐dependent in T. molitor. [ABSTRACT FROM AUTHOR]

Objectives: Periodontitis is a highly prevalent chronic inflammatory disease that results in destruction of tooth‐supporting structures followed by tooth‐loss. Until now, periodontitis has been regarded to be initiated by bacterial infection followed by aberrant host response. Although increasing evidence suggests a strong association between oxidative stress and periodontitis, precise molecular mechanism has been left unanswered. In this study, we investigated roles of SOD2, the main antioxidant enzyme maintaining reactive oxygen species (ROS) homeostasis, under inflammatory conditions. Methods: We computationally analyzed SOD2 expression in periodontitis. To confirm this data, immunoblot assay was performed with samples from periodontitis patients. The cellular mechanism of change in SOD2 expression was identified through immunoblot assay and immunofluorescence. To evaluate the molecular function of SOD2, we generated SOD2‐deficient cells by utilizing the CRISPR/Cas9 system. Results: We first determined that SOD2 expression was significantly increased in periodontitis. We also confirmed that SOD2 expression was upregulated through the NF‐κB pathway when the inflammatory signal was stronger and extended. Gene manipulation against SOD2 through the CRISPR/Cas9 system showed that the absence of SOD2 increased production of NLRP3 inflammasome components. Conclusions: Our study demonstrates that intracellular SOD2 has a protective role by suppressing NLRP inflammasome‐caspase‐1‐IL‐1β axis under inflammatory conditions. [ABSTRACT FROM AUTHOR]

Backgroud: There were limitations existing in programmed cell‐death ligand 1 (PD‐L1) as predictive biomarkers for breast cancer (BC), hence exploring the correlation between PD‐L1 levels and other biomarkers in BC may become a very useful therapeutic clinical tool. Methods: A total of 301 Chinese patients with different BC subtypes including 47 HR+/HER2+, 185 HR+/HER2−, 38 HR−/HER2+, and 31 triple‐negative breast cancer (TNBC) were enrolled in our study. Next‐generation sequencing based Yuansu450 gene panel was used for genomic alteration identification and PD‐L1 expression was tested using immunohistochemistry. Results: The most prevalent BC‐related mutations were TP53 mutations, followed by mutations in PIK3CA, ERBB2, CDK12, and GATA3 in our Chinese cohort. We found that mutations DDR2 and MYCL were only mutated in HR−/HER2+ subtype, whereas H3‐3A and NRAS mutations were only occurred in HR−/HER2− subtype. The percentage of patients with PD‐L1‐positive expression was higher in patients with HR‐/HER2− mainly due to the percentage of PD‐L1‐high level. Mutational frequencies of TP53, MYC, FAT4, PBRM1, PREX2 were observed to have significant differences among patients with different BC subtypes based on PD‐L1 levels. Moreover, a positive correlation was observed between TMB and PD‐L1 level in HR+/HER2− subtype, and showed that the proportion of patients with high PD‐L1 expression was higher than that of patients with low PD‐L1 expression in the HR+/HER2− and HR+/HER2+ cohorts with high Ki67 expression. Conclusions: The genomic alterations based on PD‐L1 and other biomarkers of different cohorts may provide more possibilities for the treatment of BC with different subtypes. [ABSTRACT FROM AUTHOR]

Targeting host factors is a complementary strategy for the development of new antiviral drugs. We screened a library of isoxazolidine and isoxazole sulfonamides and found four compounds that inhibited HIV-1 infection in human CD4+ lymphocytic T cells with no toxicity at IC90 concentrations. Structure-activity relationship showed that benzyl sulfonamides and a halo-substituted aromatic ring on the heterocycle scaffold were critical for antiretroviral activity. The size and position of the incorporated halogen had a marked effect on the antiretroviral activity. The sulfonamide derivatives had no significant effect on HIV-1 entry, reverse transcription and integration but impaired a step necessary for activation of viral gene expression. This step was Tat-independent, strongly suggesting that the target is a cell factor. A virus partially resistant to the least potent compounds could be selected but could not be propagated in the long term, consistent with the possibility that HIV-1 may be less likely to develop resistance against drugs targeting some host factors. Here, we provide evidence that novel synthetic methods can be applied to develop small molecules with antiretroviral activity that target host factors important for HIV-1 replication. [ABSTRACT FROM AUTHOR]

Abstract: Members of the claudin family play important roles in the formation of tight junctions in the kidneys, liver and intestine. Claudin-19 (Cldn19), a newly identified member of this family, is highly expressed in the kidney of the mouse. To have a better understanding on mouse claudin-19 gene expression, a 0.9-kb DNA fragment containing the 5′-flanking region of the Cldn19 gene was isolated. DNA sequence comparison between the mouse and human Cldn19 promoter regions exhibited little homology. One transcription initiation site was located at 104 nucleotides upstream of the start codon (ATG) of the Cldn19 gene. The mouse claudin-19 promoter lacked typical CAAT or GC-box. Deletion constructs of the 0.9-kb DNA fragment were generated and fused to a promoterless luciferase (Luc) reporter plasmid. Transfection studies using various kidney cell lines (MDCK, mIMCD3 and HEK293) revealed that the minimal promoter fragment resided in the −39 to −108 region, which contained a number of binding sites for transcription factors including Sp1. Site-directed mutagenesis using specific oligo probes confirmed that Sp1 was crucial for Cldn19 transactivation in the three cell lines studied. Electromobility shift assay confirmed that the nuclear extracts of these cells bound to the Sp1 oligo derived from Cldn19 promoter, but not to the mutated Sp1 oligo probe. Moreover, this DNA–protein complex would be recognized by Sp1 antibody, indicating specific Sp1 binding. Collectively, our data suggest that Sp1 binds to the claudin-19 promoter region and is responsible for its expression in the kidney cell lines in vitro. [Copyright &y& Elsevier]

The Parkinson's disease (PD)‐causative leucine‐rich repeat kinase 2 (LRRK2) belongs to the Roco family of G‐proteins comprising a Ras‐of‐complex (Roc) domain followed by a C‐terminal of Roc (COR) domain in tandem (called Roc‐COR domain). Two prokaryotic Roc‐COR domains have been characterized as 'G proteins activated by guanine nucleotide‐dependent dimerization' (GADs), which require dimerization for activation of their GTPase activity and bind guanine nucleotides with relatively low affinities. Additionally, LRRK2 Roc domain in isolation binds guanine nucleotides with relatively low affinities. As such, LRRK2 GTPase domain was predicted to be a GAD. Herein, we describe the design and high‐level expression of human LRRK2 Roc‐COR domain (LRRK2 Roc‐COR). Biochemical analyses of LRRK2 Roc‐COR reveal that it forms homodimers, with the C‐terminal portion of COR mediating its dimerization. Furthermore, it co‐purifies and binds Mg2+GTP/GDP at 1 : 1 stoichiometry, and it hydrolyzes GTP with Km and kcat of 22 nM and 4.70 × 10−4 min−1, respectively. Thus, even though LRRK2 Roc‐COR forms GAD‐like homodimers, it exhibits conventional Ras‐like GTPase properties, with high‐affinity binding of Mg2+‐GTP/GDP and low intrinsic catalytic activity. The PD‐causative Y1699C mutation mapped to the COR domain was previously reported to reduce the GTPase activity of full‐length LRRK2. In contrast, this mutation induces no change in the GTPase activity, and only slight perturbations in the secondary structure contents of LRRK2 Roc‐COR. As this mutation does not directly affect the GTPase activity of the isolated Roc‐COR tandem, it is possible that the effects of this mutation on full‐length LRRK2 occur via other functional domains. Open Practices: Open Science: This manuscript was awarded with the Open Materials Badge. For more information see: https://cos.io/our-services/open-science-badges/ The Roc‐COR GTPase supradomain of the Parkinson's disease‐causative kinase LRRK2 (LRRK2 Roc‐COR) consists of a Roc domain and a COR domain arranged in tandem. We generated the recombinant LRRK2 Roc‐COR (inset), which forms homodimers in solution. The C‐terminal portion of COR domain was found to mediate dimerization. Like conventional Ras‐like GTPases, LRRK2 Roc‐COR exhibits tight binding of Mg2+‐guanine nucleotides (inset) and low catalytic efficiency, suggesting that its proper functioning requires a guanine nucleotide exchange factor (GEF) and a GTPase‐activating protein (GAP). LRRK2 Roc‐COR is a useful tool for future investigation to identify GEF(s), GAP(s), and downstream effectors of LRRK2. [ABSTRACT FROM AUTHOR]

Objective: The basis for clinical variation related to underlying progressive supranuclear palsy (PSP) pathology is unknown. We performed a genome-wide association study (GWAS) to identify genetic determinants of PSP phenotype.Methods: Two independent pathological and clinically diagnosed PSP cohorts were genotyped and phenotyped to create Richardson syndrome (RS) and non-RS groups. We carried out separate logistic regression GWASs to compare RS and non-RS groups and then combined datasets to carry out a whole cohort analysis (RS = 367, non-RS = 130). We validated our findings in a third cohort by referring to data from 100 deeply phenotyped cases from a recent GWAS. We assessed the expression/coexpression patterns of our identified genes and used our data to carry out gene-based association testing.Results: Our lead single nucleotide polymorphism (SNP), rs564309, showed an association signal in both cohorts, reaching genome-wide significance in our whole cohort analysis (odds ratio = 5.5, 95% confidence interval = 3.2-10.0, p = 1.7 × 10-9 ). rs564309 is an intronic variant of the tripartite motif-containing protein 11 (TRIM11) gene, a component of the ubiquitin proteasome system (UPS). In our third cohort, minor allele frequencies of surrogate SNPs in high linkage disequilibrium with rs564309 replicated our findings. Gene-based association testing confirmed an association signal at TRIM11. We found that TRIM11 is predominantly expressed neuronally, in the cerebellum and basal ganglia.Interpretation: Our study suggests that the TRIM11 locus is a genetic modifier of PSP phenotype and potentially adds further evidence for the UPS having a key role in tau pathology, therefore representing a target for disease-modifying therapies. Ann Neurol 2018;84:485-496. [ABSTRACT FROM AUTHOR]

Background: Low‐dose computed tomography can identify smaller nodules more often than chest radiography in lung screening. However, complications from invasive diagnostic procedures performed to detect nodules are common. Exosomes contain a diverse array of biomolecules that reflect the biological state of the cell from which they are released. The aim of this study was to investigate the diagnostic value of bronchoalveolar lavage (BAL) fluid exosomal microRNAs (miRNAs) for early‐stage lung adenocarcinoma. Methods: We evaluated miRNAs (miR‐7, miR‐21, miR‐126, Let‐7a, miR‐17, and miR‐19) known to have diagnostic value for lung adenocarcinoma. Exosomes were isolated from the BAL fluid of control subjects (n = 15) and patients with lung adenocarcinoma (n = 13). Exosomal miRNA was analyzed using a commercial kit containing probes targeting six selected miRNAs. Results were validated via quantitative PCR. Results: The presence of miRNAs was confirmed in exosomes from BAL fluid of both lung adenocarcinoma patients and control subjects. miR‐126 (P < 0.001) and Let‐7a (P = 0.015) levels were significantly higher in the BAL fluid of lung adenocarcinoma patients than in control subjects. The BAL fluid miRNA signature was confirmed using an independent set of paired adenocarcinoma and normal tissue samples (n = 4). Lung adenocarcinoma tissues showed increased expression of miR‐126 (P = 0.039) compared to normal tissue samples. Conclusion: We identified a close correlation between BAL fluid exosomal miRNAs and tumor miRNAs. BAL fluid exosomal miRNAs obtained through noninvasive methods could serve as diagnostic biomarkers in early‐stage lung adenocarcinoma. [ABSTRACT FROM AUTHOR]

The expression and localization of the proline-rich membrane anchor (PRiMA), an anchoring protein of tetrameric globular form acetylcholinesterase (G4 AChE), were studied at vertebrate neuromuscular junctions. Both muscle and motor neuron contributed to this synaptic expression pattern. During the development of rat muscles, the expression of PRiMA and AChET and the enzymatic activity increased dramatically; however, the proportion of G4 AChE decreased. G4 AChE in muscle was recognized specifically by a PRiMA antibody, indicating the association of this enzyme with PRiMA. Using western blot and ELISA, both PRiMA protein and PRiMA-linked G4 AChE were found to be present in large amounts in fast-twitch muscle (e.g. tibialis), but in relatively low abundance in slow-twitch muscle (e.g. soleus). These results indicate that the expression level of PRiMA-linked G4 AChE depends on muscle fiber type. In parallel, the expression of PRiMA, AChET and G4 AChE also increased in the spinal cord during development. Such expression in motor neurons contributed to the synaptic localization of G4 AChE. After denervation, the expression of PRiMA, AChET and G4 AChE decreased markedly in the spinal cord, and in fast- and slow-twitch muscles. [ABSTRACT FROM AUTHOR]

Bone adapts to its environment. Osteoblasts and osteocytes are subject to mechanical load in vivo. It has been shown that osteoblasts alter cytokine expression in response to mechanical loading. However, signal transduction pathways that mediate bone cell response to mechanical stimuli have not been elucidated. In this study, we report an increase in proinflammatory gene expression in response to fluid shear stress (FSS) in human mesenchymal stem cells (hMSCs) and mouse preosteoblasts. Fluid shear stress (FSS)-induced effect was blocked by the inhibition of the calcineurin–NFATc1 axis, thus implicating a role for nuclear factor of activated T cells (NFAT) signaling in mechanotransduction. [ABSTRACT FROM AUTHOR]

Abstract: MSN1 is a putative yeast transcriptional activator involved in chromium (Cr) accumulation. Here we show that overexpression of MSN1 enhances Cr and sulfur accumulation and Cr tolerance in transgenic tobacco. In addition, we found that expression of NtST1 (Nicotiana tabacum sulfate transporter 1) was elevated in MSN1- expressing transgenic tobacco, suggesting that chromate and sulfate are taken up via the sulfate transporter in plants. Supporting this, expression of NtST1 increased levels of Cr and S in Saccharomyces cerevisiae. Our findings suggest that yeast transcriptional activators can be used for developing effective metal remediators, and for improving the nutritional status of plants. [Copyright &y& Elsevier]

Presents a study on the effect of STAT3 on hypoxia-inducible factor-1 (HIF-1)-mediated vascular endothelial factor gene expression in human renal carcinoma cells. Activation of STAT3 by hypoxia; Relationship between HIF-1α and STAT3; Result of the coimmunoprecipitation assays.