Search

Two new isocoumarins named peniciisocoumarins I and J (1 − 2), together with four known analogues (3 − 6), were obtained from Penicillium sp. GXIMD 03001, an endophytic fungus derived from the rhizophoraceous mangrove Kandelia candel. The structures and absolute configurations of 1 and 2 were determined by detailed analysis of their NMR, MS spectroscopic data, a modified Mosher's method and electronic circular dichroism data. All isolated isocoumarins were evaluated for their cytotoxic activity against four human cancer cell lines (SPCA-1, BEL-7402, GSC-7901 and MCF-7). However, none of the compounds exhibited any appreciable activity (IC 50 > 50 μ M). [Display omitted] • Two new isocoumarins, peniciisocoumarins I–J (1 and 2) were obtained from Penicillium sp. GXIMD 03001. • The absolute configuration of 1 was determined by a modified Mosher's ester method. • Cytotoxicity of all isolates against SPCA-1, BEL-7402, GSC-7901 and MCF-7 were examined. [ABSTRACT FROM AUTHOR]

Objective To identify the predictors for successful neurodynamic management in patients with patellofemoral pain syndrome. Design Prospective cohort, prediction rule study. Setting Hospital. Participants Patients with patellofemoral pain syndrome (N=51) underwent clinical examination and measurement of physical parameters, including femoral slump test, lower-extremity alignment, flexibility and muscle strength, and functional level. Intervention Patients received 6 treatment sessions of femoral nerve mobilization within 2 weeks. Main Outcome Measures Pain level during functional testing was assessed before and after the first and sixth session of treatment. Patients were then grouped into responder and nonresponder groups. Criteria for the responder group was a pain score decrease ≥50% or Global Rating Scale score ≥4. Chi-square and independent t tests were used to identify potential variables with a significance level of .10, and stepwise logistic regression was used to find predictors with a significance level of .05. Results Twenty-five patients responded to the initial treatment (immediate effect), and 28 patients responded after 6 sessions (longer-term effect). A positive femoral slump test was identified as the predictor for the immediate treatment effect. The prediction factors for the longer-term effect included responding to femoral nerve mobilization the first time and a bilateral difference in hip extension angles. Application of the clinical predictors improved the success rate to 90% for 1 treatment session and 93% for 6 treatment sessions. Conclusions Clinicians could use the positive femoral slump test and a bilateral difference in hip extension angles during the femoral slump test to determine whether or not patients with patellofemoral pain syndrome might benefit from femoral nerve mobilization. [ABSTRACT FROM AUTHOR]

PTEN is known as a tumor suppressor and plays essential roles in brain development. Here, we report that PTEN in primary sensory neurons is involved in processing itch and thermal information in adult mice. Deletion of PTEN in the dorsal root ganglia (DRG) is achieved in adult Drg11 -CreER: PTEN flox/flox (PTEN CKO) mice with oral administration of tamoxifen, and CKO mice develop pathological itch and elevated itch responses on exposure to various pruritogens. PTEN deletion leads to ectopic expression of TRPV1 and MrgprA3 in IB4+ non-peptidergic DRG neurons, and the TRPV1 is responsive to capsaicin. Importantly, the elevated itch responses are no longer present in Drg11 -CreER: PTEN flox/flox: TRPV1 flox/flox (PTEN : TRPV1 dCKO) mice. In addition, thermal stimulation is enhanced in PTEN CKO mice but blunted in dCKO mice. PTEN-involved regulation of itch-related gene expression in DRG neurons provides insights for understanding molecular mechanism of itch and thermal sensation at the spinal level. [Display omitted] • PTEN is mainly expressed in IB4+ non-peptidergic dorsal root ganglion (DRG) neurons • Adult deletion of PTEN in DRG leads to augmented itch and enhanced thermal nociception • TRPV1+ neurons are significantly increased in the DRG of PTEN CKO mice • Ectopic TRPV1 expression in DRG neurons causes itch and thermal responses in PTEN CKO Hu et al. report that PTEN, a tumor-suppressor gene, is involved in processing itch and thermal information in adult mice. They reveal that PTEN also suppresses expression of itch-related genes in primary sensory neurons and when defective leads to elevated itch and thermal sensation at the spinal level. [ABSTRACT FROM AUTHOR]

Intestinal microbiota has increasingly been recognized as a major determinant of host health due to its critical role in the maintenance of immune homeostasis. An experimental fish model with disrupted intestinal microbiota allows us to identify potential strategies for restoring or modulating microbiota composition. The aim of this study was to develop a reliable grass carp model with disrupted intestinal microbiota by treatment with an antibiotic cocktail consisting of vancomycin, enrofloxacin, florfenicol, and metronidazole at different concentrations. To identify the appropriate concentration and exposure time of antibiotic treatment required to disrupt intestinal microbiota, five antibiotic regimens were compared in terms of their effects on intestinal microbiota composition, growth and health status, and histological changes in the intestine, hepatopancreas, and spleen. Antibiotic-induced changes in the expression of inflammation- and apoptosis-related genes in grass carp intestines were also examined. The results revealed that the highest antibiotic concentration over a 28 day period was required to disrupt both luminal and mucosal microbiota through significant expansion of Proteobacteria and suppression of Fusobacteria. Treatment with this regimen induced significant down-regulation of caspase-8, MLCK, ZO-1, IL-17N, and IL-23R, and up-regulation of IL-1β and IL-6 in the intestine, suggesting possible effects on intestinal epithelial tight junction and inflammatory conditions. This treatment did not produce substantial histopathological changes, and was not directly detrimental to growth and health. Therefore, this antibiotic regimen may provide a suitable option to disrupt intestinal microbiota in grass carp. In addition, our study also implies that fish may serve as a promising model for investigating growth-promoting effects of antibiotics, when treated with antibiotics in low doses. • Antibiotic treatment did not affect alpha diversity of the luminal microbiota. • Antibiotic exposure duration matters more than concentration in disrupting microbiota. • The microbiota composition differed substantially between intestinal content and mucosa. • The growth-promoting effect of low-dose antibiotics was confirmed in grass carp. [ABSTRACT FROM AUTHOR]

Abstract: Taxonomical analysis of two genetically distinguished Lactobacillus strains isolated from traditional Chinese fermented vegetables ‘Suan cai’ was performed. They formed l-lactate from glucose, were facultatively heterofermentative, and had a DNA G+C content of 53–54mol%. They fermented d- and l-arabinose. They produced lactate, ethanol and acetate from gluconate at a molar ratio of 1.1:0.4:0.7. Phylogenetic analysis of 16S rDNA revealed that the two strains were closely related to L. perolens. DNA–DNA hybridization analysis revealed that the two strains were different from L. perolens type strain DSM 12744 and formed a separate cluster with L. perolens DSM 12745. G+C molar content of DNA of the former is 51%, whereas those of the latter strains were in the range of 53–54%. Based on the results, we propose that the new species be named L. harbinensis sp. nov. and that L. perolens DSM 12745 be reclassified as L. harbinensis DSM 12745. The type strain of L. harbinensis DSM 16991T (=AHU 1762T=SBT 10908T). [Copyright &y& Elsevier]

[Display omitted] • 20E stimulates the expression of LdFoxO. • Silencing LdFoxO represses 20E signaling and reduces larval weight. • RNAi of LdFoxO impairs larva-pupa-adult transition. • 20E or JH consumption cannot restore defects in LdFoxO depleted animals. Forkhead box O (FoxO) protein, a major downstream transcription factor of insulin/insulin-like growth factor signaling/target of rapamycin pathway (IIS/TOR), is involved in the regulation of larval growth and the determination of organ size. FoxO also interacts with 20-hydroxyecdysone (20E) and juvenile hormone (JH) signal transduction pathways, and hence is critical for larval development in holometabolans. However, whether FoxO plays a critical role during larval metamorphosis needs to be further determined in Leptinotarsa decemlineata. We found that 20E stimulated the expression of LdFoxO. RNA interference (RNAi)-aided knockdown of LdFoxO at the third-instar stage repressed 20E signaling and reduced larval weight. Although the resultant larvae survived through the third-fourth instar ecdysis, around 70% of the LdFoxO depleted moribund beetles developmentally arrested at prepupae stage. These LdFoxO depleted beetles were completely wrapped in the larval exuviae, gradually darkened and finally died. Moreover, approximately 12% of the LdFoxO RNAi beetles died as pharate adults. Ingestion of either 20E or JH by the LdFoxO depletion beetles excessively rescued the corresponding hormonal signals, but could not alleviate larval performance and restore defective phenotypes. Therefore, FoxO plays an important role in regulation of larval-pupal-adult transformation in L. decemlineata , in addition to mediation of IIS/TOR pathway and stimulation of ecdysteroidogenesis. [ABSTRACT FROM AUTHOR]

The genomic and coding sequences of four members of grass carp interleukin (IL)-17 cytokine family, IL-17A/F2, IL-17A/F3, IL-17C and IL-17N, were isolated using rapid amplification of cDNA ends and conventional PCR techniques. A phylogenetic analysis showed that grass carp IL-17 family genes, including previously identified IL-17A/F1 and IL-17D, clustered with their counterparts from other fish species. IL-17D had two exons and one intron, while all the other members were composed of three exons and two introns. A multiple sequence alignment revealed that grass carp IL-17 family cytokines shared a cystine-knot motif that was highly conserved in mammals. A quantitative real-time PCR analysis revealed that these genes were expressed in the different examined tissues at varying levels. Moreover, they displayed different tissue expression patterns in fish at 24 h after intraperitoneal injection (IP) or anal intubation (AI) of Aeromonas hydrophila , implying roles in inflammatory responses. Next, we compared disease activity status and histopathological changes in intestinal tissues over a 7 day period after IP or AI, and then examine the potential differential effects of different infection routes on the expression profiles of IL-17 family genes in inflamed intestinal tissues. The intraperitoneal route resulted in more pronounced pathological symptoms than the anal route in terms of the disease activity index (DAI). Changes in the DAI highly coincided with the severity of intestinal inflammation. The two infection routes resulted in differential IL-17 family gene expression patterns over time in grass carp intestine, which suggests that IL-17 family members may play differential roles in mucosal and systemic immune responses and that IL-17N and IL-17A/F1 might possibly function as pro-inflammatory cytokines that provide defense against bacterial infection in grass carp intestine. • Grass carp IL-17A/F2, IL-17A/F3, IL-17C and IL-17 N sequences were isolated. • Most grass carp IL-17 family genes exhibited an identical exon-intron structure. • Grass carp IL-17 family cytokines contained a cystine-knot motif found in mammals. • Tissue expression patterns of IL-17 family genes differed after bacterial infection. • IL-17 family members had unique roles in mucosal and systemic immune responses. [ABSTRACT FROM AUTHOR]

The RNA exosome is a ribonuclease complex that mediates both RNA processing and degradation. This complex is evolutionarily conserved, ubiquitously expressed, and required for fundamental cellular functions, including rRNA processing. The RNA exosome plays roles in regulating gene expression and protecting the genome, including modulating the accu-mulation of RNA-DNA hybrids (R-loops). The function of the RNA exosome is facilitated by cofactors, such as the RNA helicase MTR4, which binds/remodels RNAs. Recently, missense mutations in RNA exosome subunit genes have been linked to neurological diseases. One possibility to explain why missense mutations in genes encoding RNA exosome subunits lead to neurological diseases is that the complex may interact with cell- or tissue-specific cofactors that are impacted by these changes. To begin addressing this question, we performed immunoprecipitation of the RNA exosome subunit, EXOSC3, in a neuronal cell line (N2A), followed by proteomic analyses to identify novel interactors. We identified the putative RNA helicase, DDX1, as an interactor. DDX1 plays roles in doublestrand break repair, rRNA processing, and R-loop modulation. To explore the functional connections between EXOSC3 and DDX1, we examined the interaction following doublestrand breaks and analyzed changes in R-loops in N2A cells depleted for EXOSC3 or DDX1 by DNA/RNA immunoprecipitation followed by sequencing. We find that EXOSC3 interaction with DDX1 is decreased in the presence of DNA damage and that loss of EXOSC3 or DDX1 alters R-loops. These results suggest EXOSC3 and DDX1 interact during events of cellular homeostasis and potentially suppress unscrupulous expression of genes promoting neuronal projection. [ABSTRACT FROM AUTHOR]

Grim-19 (gene associated with retinoid-IFN-induced mortality 19), the essential component of complex I of mitochondrial respiratory chain, functions as a noncanonical tumor suppressor by controlling apoptosis and energy metabolism. However, additional biological actions of Grim-19 have been recently suggested in male reproduction. We investigated here the expression and functional role of Grim-19 in murine testis. Testicular Grim-19 expression was detected from mouse puberty and increased progressively thereafter, and GRIM-19 protein was observed to be expressed exclusively in interstitial Leydig cells (LCs), with a prominent mitochondrial localization. In vivo lentiviral vector-mediated knockdown of Grim-19 resulted in a significant decrease in testosterone production and triggered aberrant oxidative stress in testis, thus impairing male fertility by inducing germ cell apoptosis and oligozoospermia. The control of testicular steroidogenesis by GRIM-19 was validated using the in vivo knockdown model with isolated primary LCs and in vitro experiments with MA-10 mouse Leydig tumor cells. Mechanistically, we suggest that the negative regulation exerted by GRIM-19 deficiency-induced oxidative stress on steroidogenesis may be the result of two phenomena: a direct effect through inhibition of phosphorylation of steroidogenic acute regulatory protein (StAR) and subsequent impediment to StAR localization in mitochondria and an indirect pathway that is to facilitate the inhibiting role exerted by the extracellular matrix on the steroidogenic capacity of LCs via promotion of integrin activation. Altogether, our observations suggest that Grim-19 plays a potent role in testicular steroidogenesis and that its alterations may contribute to testosterone deficiency-related disorders linked to metabolic stress and male infertility. [ABSTRACT FROM AUTHOR]

Protein arginine methyltransferase 5 (PRMT5) is a member of the arginine methyltransferase protein family that critically mediates the symmetric dimethylation of Arg-3 at histone H4 (H4R3me2s) and is involved in many key cellular processes, including hematopoiesis. However, the post-translational modifications (PTMs) of PRMT5 that may affect its biological functions remain less well-understood. In this study, using MS analyses, we found that PRMT5 itself is methylated in human erythroleukemia Lys-562 cells. Biochemical assays revealed that coactivator-associated arginine methyltransferase 1 (CARM1) interacts directly with and methylates PRMT5 at Arg-505 both in vivo and in vitro. Substitutions at Arg-505 significantly reduced PRMT5's methyltransferase activity, decreased H4R3me2s enrichment at the γ-globin gene promoter, and increased the expression of the γ-globin gene in Lys-562 cells. Moreover, CARM1 knockdown consistently reduced PRMT5 activity and activated γ-globin gene expression. Importantly, we show that CARM1-mediated methylation of PRMT5 is essential for the intracellular homodimerization ofPRMT5to its active form. These results thus reveal a critical PTM of PRMT5 that represses human γ-globin gene expression.Weconclude that CARM1-mediated asymmetric methylation of PRMT5 is critical for its dimerization and methyltransferase activity leading to the repression of γ-globin expression. Given PRMT5's crucial role in diverse cellular processes, these findings may inform strategies for manipulating its methyltransferase activity for managing hemoglobinopathy or cancer. [ABSTRACT FROM AUTHOR]

Winter jujube ( Zizyphus jujuba Mill.) is a kind of high quality, fresh jujube fruit originated from China. Selenium (Se) affects the chemical composition and antioxidative properties in plants. At appropriate concentrations, Se acts as an antioxidant or activates mechanisms that can alleviate oxidative stress in the chloroplast, and enhances plant growth. Although the addition of Se is known to increase Se uptake in crops, it is unclear whether exogenous Se affects nutritional and functional components in winter jujube. Winter jujube trees were sprayed with selenium (as sodium selenite) at five concentrations (0, 25, 50, 100, or 200 mg/L of Se) three times at the fruitlet stage between July and August. The Se content, ratio of organic Se to total Se in leaves and fruits, fruit quality, and yield were measured. At Se concentrations of 50 mg/L or higher, total Se content increased rapidly whereas the ratio increased initially and decreased later. Different Se concentrations affected the quality of fruits differently: at 50 mg/L, compared to their values in the unsprayed control trees, vitamin C content was greater by 20.94%; soluble sugars, 29.48%; total flavonoids, 43.48%; and the sugar: acid ratio, 41.81%. Along with the increase of Se addition, average fruit weight and yield of winter jujube presented a trend of increasing up to 50 mg L −1 and decreasing thereafter. Thus, the foliar spray of sodium selenite at Se concentration of 50 mg/L is recommended during the fruitlet stage of Zhanhua winter jujube. [ABSTRACT FROM AUTHOR]

Production of hemoglobin during development is tightly regulated. For example, expression from the human β-globin gene locus, comprising β-, δ-, ∊-, and γ-globin genes, switches from ∊-globin to γ-globin during embryonic development and then from γ-globin to β-globin after birth. Expression of human ∊-globin in mice has been shown to ameliorate anemia caused by β-globin mutations, including those causing β-thalassemia and sickle cell disease, raising the prospect that reactivation of ∊-globin expression could be used in managing these conditions in humans. Although the human globin genes are known to be regulated by a variety of multiprotein complexes containing enzymes that catalyze epigenetic modifications, the exact mechanisms controlling ∊-globin gene silencing remain elusive. Here we found that the heterochromatin protein HP1γ, a multifunctional chromatin- and DNA-binding protein with roles in transcriptional activation and elongation, represses ∊-globin expression by interacting with a histone-modifying enzyme, lysine methyltransferase SUV4-20h2. Silencing of HP1γ expression markedly decreased repressive histone marks and the multimethylation of histone H3 lysine 9 and H4 lysine 20, leading to a significant decrease in DNA methylation at the proximal promoter of the ∊-globin gene and greatly increased ∊-globin expression. In addition, using chromatin immunoprecipitation, we showed that SUV4-20h2 facilitates the deposition of HP1γ on the ∊-globin-proximal promoter. Thus, these data indicate that HP1γ is a novel epigenetic repressor of ∊-globin gene expression and provide a potential strategy for targeted therapies for β-thalassemia and sickle cell disease. [ABSTRACT FROM AUTHOR]

The T-synthase (Core 1 β3-galactosyltransferase) and its molecular chaperone Cosmc regulate the biosynthesis of mucin type O-glycans on glycoproteins, and evidence suggests that both T-synthase and Cosmc are transcriptionally suppressed in several human diseases, although the transcriptional regulation of these two genes is not understood. Here we characterized the promoters essential for human Cosmc and T-synthase transcription. The upstream regions of the genes lack a conventional TATA box but contain CpG-islands: cCpG-I and cCpG-II for Cosmc and tCpG for T-synthase. Using luciferase reporter assays, site-directed mutagenesis, ChIP assays, and Mithramycin A treatment, we identified the core promoters within cCpG-II and tCpG, which contain two binding sites for Kruppel-like transcription factors (KLF), including SP1/SP3, respectively. Methylome analysis of Tn4 B cells, which harbor a silenced Cosmc, confirmed the hypermethylation of Cosmc core promoter, but not for that of T-synthase. These results demonstrate that Cosmc and T-synthase are transcriptionally regulated at a basal level by SP/KLF family transcription factors, which explains their ubiquitous and coordinated expression, and also indicates that they are differentially epigenetically regulated beyond X-chromosome imprinting. These results are important in understanding the regulation of these genes that have roles in human diseases, such as IgA Nephropathy and cancer. [ABSTRACT FROM AUTHOR]