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Background and objective: In recent years, carbapenemase-producing Enterobacterales (CPE) resistance to antibiotics has dramatically increased leading to limitations of their treatment options. In the present study, we investigated the occurrence of carbapenemase-producing Escherichia coli and Salmonella in rural Burkina Faso.Materials and methods: Salmonella isolates were serotyped according to the Kauffman White scheme. Diarrheagenic Escherichia coli (DEC) strains was identified using 16-plex Polymerase Chain Reaction (PCR), whereas antibiotic susceptibility was realized using the disk diffusion method. Furthermore, multiplex PCR assays were carried out using oligonucleotides to detect the presence of genes of the blaKPC, blaVIM, blaIMP, blaTEM, blaSHV, blaOXA and blaCTX-M types in all E. coli and Salmonella strains.Results: The study highlighted high resistance rates of the identified bacteria to common antibiotics. Likewise, two strains of E. coli were imipenem resistant with carbapenemase-encoding genes. The genes detected were Klebsiella pneumoniae carbapenemase (KPC), Verona integrin-encoded metallo-β-lactamase (VIM) and Imipenemase (IMP-2) reaching a rate of 40% each in E. coli strains. However, no Salmonella carbapenemases blaKPC, blaVIM or blaIMP were detected.Conclusion: This study showed that for a real-time infection control and prompt application of antimicrobial chemotherapy, characterization of carbapenemase-producing Enterobacterales in patients is crucial.Keywords: Antibiotics, Carbapenemase-Producing Enterobacterales, children, Burkina Faso

Bioenergy recovery from cashew nut shells was investigated throughout using efficient inoculums source and appropriate pretreatment. Physicochemical characteristics of shells and inoculums including pH, volatile fatty acid (VFA, total solid, volatile solid, ash were determined using standard methods. Total anaerobes and methanogenic archaea from inocula were determined by MPP method. Wastewater (WW), sludge from bioreactor (SBR), bovine dung (BD) and mixed inoculums (MIX) were used to evaluate inoculums source in batch system. Biochemical methane potential of pretreated shells was evaluated. Biogas was measurement by liquid displacement. CH4 and CO2 were performed by GC. MIX and WW showed high concentration of methanogenic bacteria (2.3 102 CFU/mL). Best biomethane levels 70.38% with yields of 55.52 L biogas. (Kg VS)-1 was observed with old shells inoculated by MIX. Low productivity was noted with thermal and biological treatment of old and fresh shells, 11.20 and 0.02 L CH4. (Kg VS)-1, respectively. Inoculums source has significant effect on biogas production. Mixed inoculums exhibited significantly high yields of biomethane. Thermo-biological pretreatment seems to be not appropriate for a better biomethane production. Combination of thermochemical and biological pretreatment could be necessary for best biomethane production yield.

Fibroblast growth factor receptor type 2 (FGFR2) has emerged as a key oncogenic factor that regulates gastric cancer (GC) progression, but the underlying mechanism of FGF–FGFR2 signaling pathway remains largely unknown. To identify the potential molecular mechanisms of the oncogenic FGFR2 in gastric carcinogenesis and convey a novel therapeutic strategy, we profiled the FGFR alterations and analyzed their clinical associations in TCGA and Hong Kong GC cohorts. We found that FGFR2 overexpression in GC cell lines and primary tumors predicted poor survival and was associated with advanced stages of GC. Functionally, growth abilities and cell cycle progression of GC were inhibited by inactivation of ERK–MAPK signal transduction after FGFR2 knockdown, while apoptosis was promoted. Meanwhile, the first-line anti-cancer drug sensitivity was enhanced. RNA-seq analysis further revealed that YAP1 signaling serves as a significant downstream modulator and mediates the oncogenic signaling of FGFR2. When stimulating FGFR2 by rhFGF18, we observed intensified F-actin, nuclear accumulation of YAP1, and overexpression of YAP1 targets, but these effects were attenuated by either FGFR2 depletion or AZD4547 administration. Additionally, the FGF18–FGFR2 signaling upregulated YAP1 expression through activating c-Jun, an effector of MAPK signaling. In our cohort, 28.94% of GC cases were characterized as FGFR2, c-Jun, and YAP1 co-positive and demonstrated worse clinical outcomes. Remarkably, we also found that co-targeting FGFR2 and YAP1 by AZD4547 and Verteporfin synergistically enhanced the antitumor effects in vitro and in vivo. In conclusion, we have identified the oncogenic FGF–FGFR2 regulates YAP1 signaling in GC. The findings also highlight the translational potential of FGFR2–c-Jun–YAP1 axis, which may serve as a prognostic biomarker and therapeutic target for GC.

Hippo signaling functions to limit cellular growth, but the aberrant nuclear accumulation of its downstream YAP1 leads to carcinogenesis. YAP1/TEAD complex activates the oncogenic downstream transcription, such as CTGF and c-Myc. How YAP1 is protected in the cytoplasm from ubiquitin-mediated degradation remains elusive. In this study, a member of Angiomotin (Motin) family, AMOTL1 (Angiomotin Like 1), was screened out as the only one to promote YAP1 nuclear accumulation by several clinical cohorts, which was further confirmed by the cellular functional assays. The interaction between YAP1 and AMOTL1 was suggested by co-immunoprecipitation and immunofluorescent staining. The clinical significance of the AMOTL1–YAP1–CTGF axis in gastric cancer (GC) was analyzed by multiple clinical cohorts. Moreover, the therapeutic effect of targeting the oncogenic axis was appraised by drug-sensitivity tests and xenograft-formation assays. The upregulation of AMOTL1 is associated with unfavorable clinical outcomes of GC, and knocking down AMOTL1 impairs its oncogenic properties. The cytoplasmic interaction between AMOTL1 and YAP1 protects each other from ubiquitin-mediated degradation. AMOTL1 promotes YAP1 translocation into the nuclei to activate the downstream expression, such as CTGF. Knocking down AMOTL1, YAP1, and CTGF enhances the therapeutic efficacies of the first-line anticancer drugs. Taken together, AMOTL1 plays an oncogenic role in gastric carcinogenesis through interacting with YAP1 and promoting its nuclear accumulation. A combination of AMOTL1, YAP1, and CTGF expression might serve as a surrogate of Hippo activation status. The co-activation of the AMOTL1/YAP1–CTGF axis is associated with poor clinical outcomes of GC patients, and targeting this oncogenic axis may enhance the chemotherapeutic effects.

Germline mutation in the PTEN gene is the genetic basis of PTEN hamartoma tumor syndrome with the affected individuals harboring features of autism spectrum disorders. Characterizing a panel of 14 autism‐associated PTEN missense mutations revealed reduced protein stability, catalytic activity, and subcellular distribution. Nine out of 14 (64%) PTEN missense mutants had reduced protein expression with most mutations confined to the C2 domain. Selected mutants displayed enhanced polyubiquitination and shortened protein half‐life, but that did not appear to involve the polyubiquitination sites at lysine residues at codon 13 or 289. Analyzing their intrinsic lipid phosphatase activities revealed that 78% (11 out of 14) of these mutants had twofold to 10‐fold reduction in catalytic activity toward phosphatidylinositol phosphate substrates. Analyzing the subcellular localization of the PTEN missense mutants showed that 64% (nine out of 14) had altered nuclear‐to‐cytosol ratios with four mutants (G44D, H123Q, E157G, and D326N) showing greater nuclear localization. The E157G mutant was knocked‐in to an induced pluripotent stem cell line and recapitulated a similar nuclear targeting preference. Furthermore, iPSCs expressing the E157G mutant were more proliferative at the neural progenitor cell stage but exhibited more extensive dendritic outgrowth. In summary, the combination of biological changes in PTEN is expected to contribute to the behavioral and cellular features of this neurodevelopmental disorder., Mutation of the PTEN gene has been linked to autism spectrum disorders (ASDs) and other disorders with autism‐like features. Here, Andrew Chen and co‐authors characterized a panel of PTEN autism‐associated mutants, unearthing a E157G PTEN mutant that displays preferential localization to the cell nucleus. The E157G mutant was knocked‐in to an iPSC line that recapitulated a similar nuclear‐targeting preference. In addition, iPSCs expressing the E157G mutant were more proliferative at the neural progenitor cell stage and exhibited more extensive dendritic outgrowth when induced to undergo neuronal differentiation, hinting at the contribution of PTEN to the cellular features of ASDs.

With high annual mortality rates among young children, antimicrobial resistant salmonellosis is considered a major public health concern worldwide. Antimicrobial resistant salmonellosis is a worldwide health issue, particularly in low income countries with high microbially-derived food contaminations. As a result, it is important to better understand the biological factors that may control these bacteria’s dissemination low immunity individuals such as children. Thus, a sound epidemiological surveillance and control of salmonellosis (that is, tet and catA1) requires a better understanding of the role that class 1, 2 and 3 integrons play in the spread of these antimicrobial resistant genes. A total of 275 stool samples of children suffering of diarrhea in rural Burkina Faso were collected and their Salmonella species were screened. The antimicrobial resistance determinants were investigated by Polymerase Chain Reaction, checking the presence of class 1, 2, 3 integrons, tet and catA1 resistance genes. Seven of the nine confirmed Salmonella strains (78%) were multidrug resistant while 100% were resistant to amoxicillin. Antibiotic resistance genes catA and tet were present in 11.1 and 22.2%, respectively. Integrons were detected as follows: Int1 (44.4%) and Int2 (22.2%). No class 3 integron was detected. A surveillance and control programme of antimicrobial drug resistant Salmonella species is of paramount importance for limiting spread of these pathogens among children. Key words: Antibiotic resistance genes, Class 1 and 2 integrons, Salmonella, children.

Acetic acid bacteria are known for their ability to oxidize ethanol to acetic acid. This study investigated to identify dominant strain acetic acid bacteria involving in fermented juice of mango and capable to produce Vinegar, total of 4 bacteria (CRSBAN-BVA1, CRSBAN-BVK1, CRSBAN-BVK2, CRSBAN-BVI1) bacteria strains were preselected ¶for the analyses. The strains were examined with biochemical, physiological and phenotypical methods such as Gram die, catalase and oxidase test, ethanol oxidation to acetic acid, and over to CO2 and H2O and also metabolism of carbohydrate was tested, for their affiliation to the genera of acetic acid bacteria. Subsequently, genotypic identification was conducted by sequencing the gene coding for 16S rRNA of one targeted strain and phylogenetic analysis was realized throughout 16S rRNA sequences. The results showed that one of the isolated strain (CRSBAN-BVA1) present 99. 90% of similarity in the sequence 16S rRNA region with Acetobacter tropicalis. It demonstrated that bacterial diversity in the mango vinegar is dominated by A. tropicalis. Therefore this strain is potentially useful for its utilization as a starter in vinegar production. Key words: Fermented juice, acetic acid bacteria, 16S rRNA gene sequence, Acetobacter tropicalis.

Essential oils are natural substances which can be used as natural conservators for foods. The present work aimed to evaluate biological properties of essential oil of Hyptis suaveolens and its effect on shelf-life of beef. The extraction was carried out by hydro-distillation. Antioxidant activity was determined by method of DPPH and FRAP. Antimicrobial activity was determined by the microdilution and agar diffusion methods. The sensitivity of eleven pathogen microbial strains was tested. The conservation test of ground meat using essential oil was carried out by monitoring the evolution of the microbial groups. Result showed that extraction yield of essential oil was 0.21% (m/v). The DPPH assay gave IC50 of 16.367 ± 0.0739 µL and was confirmed by the FRAP assay. The essential oil from H. suaveolens showed inhibitory activity on microbial strains. Biopreservation assay revealed a significant decrease of microbial charge during conservation time. Thus essential oil was successful in vitro and its activity on prolonging shelf-life of beef was found to be 7 days. The results of the present study demonstrated the possibility of essential oil from H. suaveolens to be used as natural conservator for food industry. Key words: Hyptis suaveolens, essential oil, biopreservation, biological properties, meat.

Background Lymphoepithelioma-like hepatocellular carcinoma (LEL-HCC) as a distinct variant of HCC displayed immunologically hot tumor features with prominent tumor-infiltrating CD8+T lymphocytes (TILs). Our whole exosome sequencing data found an increased prevalence of chromosome 11q13.3 amplification in LEL-HCC, in which contains fas-associated death domain (FADD) that displayed a strongest positive correlation with differentially expressed genes function in TIL migration. We hence aim to elucidate the functional roles and molecular mechanisms of hepatoma-intrinsic FADD in regulating TIL abundance in general HCC patients. Methods FADD-overexpressed HepG2 (FADD-oe) and FADD-knockdown huh7 cells (FADD-kd) were constructed to investigate cell growth rate by colony formation and MTS assay in vitro, subcutaneous tumor in immunodeficient nude and NOD/SCID mice in vivo. A co-culture model of human peripheral blood-derived- CD8+T cells and HepG2-oe or Huh7-kd in vitro, T cell adoptive transfer and humanized mice models in vivo were used to detect T migration preference. The downstream functional chemokines and molecular pathways controlled by FADD were determined by RT-qPCR, ELISA, western blot as well as gene modulation assays. Finally, the associations among FADD (Fadd) expression, TIL proportion and patient prognosis or tumor development were evaluated by NanoString/immunohistochemistry analysis in HCC patients or RT-qPCR/flow cytometry in orthotopic HCC mouse models. Results CD8+T cell migration rate was significantly enhanced in FADD-oe tumors, but reduced in FADD-kd tumors in both HCC cells-T cell co-culture system in vitro and subcutaneous tumor models in vivo. Mechanistically, FADD activated NF-κB, which in turn promoted T cell trafficking chemokine CCL5 production. Furthermore, Fadd expression was positively correlated with Ccl5 and TIL proportions, but negatively correlated with orthotopic HCC tumor weights. As we showed that FADD was positively correlated with CD8+T cells in HCC patients, our data pinpointed that FADD controls TIL abundance in HCC. Conclusions Our findings reveal an underlying mechanism of TIL accumulation in HCC, which provides a novel strategy of FADD activation in turning immunologically cold tumors into hot ones for better prognosis and immunotherapeutic responses.

Autosomal dominant retinitis pigmentosa (adRP) is frequently caused by mutations in RHO, the gene for rhodopsin. In previous experiments in dogs with the T4R mutation in RHO, an AAV2/5 vector expressing both an shRNA directed to human and dog RHO mRNA and an shRNA-resistant human RHO cDNA (AAV-RHO820-shRNA820) prevented retinal degeneration for more than 8 months following injection. To confirm that this same vector could protect the retinas of a different species and bearing a different RHO mutation, we injected mice transgenic for human P23H RHO at postnatal day 30 in one eye. For nine months, we monitored their retinal structure using spectral- domain optical coherence tomography (SD-OCT) and retinal function using electroretinography (ERG). We compared these to P23H RHO transgenic mice injected with AAV-GFP. Though retinas continued to thin over time, compared to control injected eyes, AAV-RHO820-shRNA820 slowed the loss of photoreceptor cells and decreased ERG amplitudes in AAV-RHO820-shRNA820 eyes during the nine-month study period. Unexpectedly, we also observed preservation of retinal structure and function in the untreated contralateral eyes of AAV-RHO820-shRNA820 treated mice. PCR analysis and western blots provided evidence that a low amount of vector from injected eyes was present in uninjected eyes.

Background The aim of this study was to determine the resistance of diarrheagenic Escherichia coli strains to β-lactams antibiotics and to perform the molecular characterization of Extended Spectrum β-lactamases (ESBL) and integrons genes. Methods This study was carried out from August 2013 to October 2015 and involved 31 DEC strains isolated from diarrheal stools samples collected from children less than five years of age. The identification and characterization of DEC strains was done through the standard biochemical tests those were confirmed using API 20E and Polymerase Chain Reaction (PCR). The determination of antimicrobial resistance was realized by the disk diffusion method then an amplification of the β-lactamase resistance genes and integrons by PCR was done. Results Out of the 419 E. coli strains identified, 31 isolates (7.4%) harbored the DEC virulence genes. From these DEC, 21 (67.7%) were ESBL-producing E. coli. Susceptibility to ESBL-producing E. coli showed that the majority of isolates were highly resistant to amoxicillin (77.4%), amoxicillin clavulanic acid (77.4%) and piperacillin (64.5%). The following antibiotic resistance genes and integron were identified from the 31 DEC isolates: blaTEM (6.5%), blaSHV (19.4%), blaOXA (38.7%) blaCTX−M (9.7%), Int1 (58.1%) and Int3 (19.4%). No class 2 integrons (Int2) was characterized. Conclusions Because of the high prevalence of multidrug-resistant ESBL organisms found in this study among pediatric patients, there is a need of stringent pediatric infection control measures.

Tumor suppressor genes (TSGs) exhibit distinct evolutionary features. We speculated that TSG promoters could have evolved specific features that facilitate their tumor-suppressing functions. We found that the promoter CpG dinucleotide frequencies of TSGs are significantly higher than that of non-cancer genes across vertebrate genomes, and positively correlated with gene expression across tissue types. The promoter CpG dinucleotide frequencies of all genes gradually increase with gene age, for which young TSGs have been subject to a stronger evolutionary pressure. Transcription-related features, namely chromatin accessibility, methylation and ZNF263-, SP1-, E2F4- and SP2-binding elements, are associated with gene expression. Moreover, higher promoter CpG dinucleotide frequencies and chromatin accessibility are positively associated with the ability of TSGs to resist downregulation during tumorigenesis. These results were successfully validated with independent datasets. In conclusion, TSGs evolved specific promoter features that optimized cancer resistance through achieving high expression in normal tissues and resistance to downregulation during tumorigenesis.

This work was supported by a Newton Fund award from the Medical Research Council (United Kingdom) MR/N019296/1 and the Philippine Council for Health Research and Development. Additional support was provided by the National Institute for Health Research (United Kingdom), the Global Health Research Unit on genomic surveillance of AMR (16/136/111) and by research grant U01CA207167 from the US National Institutes of Health. Antimicrobial-resistant Neisseria gonorrhoeae is a major threat to public health and is of particular concern in the Western Pacific Region, where the incidence of gonorrhoea is high. The Antimicrobial Resistance Surveillance Program (ARSP) has been capturing information on resistant gonorrhoea since 1996, but genomic epidemiology studies on this pathogen are lacking in the Philippines. We sequenced the whole genomes of 21 N. gonorrhoeae isolates collected in 2013-2014 by ARSP. The multilocus sequence type, multiantigen sequence type, presence of determinants of antimicrobial resistance and relatedness among the isolates were all derived from the sequence data. The concordance between phenotypic and genotypic resistance was also determined. Ten of 21 isolates were resistant to penicillin, ciprofloxacin and tetracycline, due mainly to the presence of the blaTEM gene, the S91F mutation in the gyrA gene and the tetM gene, respectively. None of the isolates was resistant to ceftriaxone or cefixime. The concordance between phenotypic and genotypic resistance was 92.38% overall for five antibiotics in four classes. Despite the small number of isolates studied, they were genetically diverse, as shown by the sequence types, the N. gonorrhoeae multiantigen sequence typing types and the tree. Comparison with global genomes placed the Philippine genomes within global lineage A and led to the identification of an international transmission route. This first genomic survey of N. gonorrhoeae isolates collected by ARSP will be used to contextualize prospective surveillance. It highlights the importance of genomic surveillance in the Western Pacific and other endemic regions for understanding the spread of drug-resistant gonorrhoea worldwide. Publisher PDF

Background & aims Immune checkpoint blockade (ICB) has been approved for treatment of hepatocellular carcinoma (HCC). However, many patients with advanced HCC are non-responders to ICB monotherapy. Cytotoxic chemotherapy has been proposed to modulate the tumor microenvironment (TME) and sensitize tumors to ICB. Thus, we aimed to study the combination of cytotoxic chemotherapy and ICB in an orthotopic HCC model. Methods Preclinical orthotopic HCC mouse models were used to elucidate the efficacy of 5-fluorouracil (5-FU) and ICB. The mice were intrahepatically injected with RIL-175 or Hepa1-6 cells, followed by treatment with 5-FU and anti-programmed cell death ligand 1 (PD-L1) antibody. Myeloid-derived suppressor cells (MDSCs) were depleted to validate their role in attenuating sensitivity to immunotherapy. Flow cytometry-based immune profiling and immunofluorescence staining were performed in mice and patient samples, respectively. Results 5-FU could induce intratumoral MDSC accumulation to counteract the infiltration of T lymphocytes and natural killer cells, thus abrogating the anti-tumor efficacy of PD-L1 blockade. In clinical samples, MDSCs accumulated and CD8+ T cell numbers decreased following transarterial chemoembolization. Conclusion 5-FU can trigger the accumulation of immunosuppressive MDSCs, impairing the response to PD-L1 blockade in HCC. Our data suggest that the combination of specific chemotherapy and ICB may impair anti-tumor immune responses, warranting further study in preclinical models and consideration in clinical settings. Lay summary Our findings suggest that some chemotherapies may impair the anti-tumor efficacy of immunotherapy. Further studies are required to uncover the specific effects of different chemotherapies on the immunological profile of tumors. This data will be critical for the rational design of combination immunotherapy strategies for patients with hepatocellular carcinoma., Graphical abstract, Highlights • Some chemotherapeutic agents could counteract the efficacy of PD-L1 blockade in HCC. • 5-FU mediates MDSC accumulation in tumors which blunts the response to immunotherapy. • MDSCs are enriched in HCC patients treated with transarterial chemoembolization.

Background The immune-checkpoint-blockade (ICB) therapy has produced promising and yet modest objective response rates in hepatocellular carcinoma (HCC), which has been attributable to the strong immunosuppressive tumor microenvironment (TME). As a key player in the TME, myeloid-derived suppressor cell (MDSC) shows potent T cell-suppressive activity that remarkably associates with poor prognosis and ICB resistance of cancer patients. While targeting MDSC can blunt T cell activity, a new approach is directed towards driving MDSC differentiation into antigen presentation cell crucial for T cell priming and activation. We have recently shown that hepatoma-intrinsic cyclin-dependent-kinase 20 (CDK20), or cell-cycle-related-kinase (CCRK) depletion diminishes MDSC-mediated immunosuppression leading to improved ICB efficacy (Gut 2018). As emerging evidence highlights the key roles of immune cell-intrinsic CDKs, we aimed to further explore the potentials of CCRK in immune cell identity. Methods The expression profile of CCRK was determined in flow-sorted immune cells from tumor-bearing mice and HCC patients. Functional significance and molecular mechanisms of CCRK in MDSCs were conducted by gene knockdown in human blood-derived MDSCs, followed by mRNA and protein detection, qChIP-PCR and multi-colour flow cytometry. The MDSC differentiation and T cell suppression in tumorigenicity were validated in HCC mouse model with intratumoral MDSC injection. Results We uncovered specific over-expression of CCRK in MDSCs but not lymphocytes from tumor-bearing mice and HCC patients. Notably, blockade of MDSC-intrinsic CCRK induced its differentiation into antigen-presenting macrophage, which amplified T cell responses in vitro and in vivo, resulting in reduced tumorigenicity. CCRK inhibition suppressed signal transducer and activator of transcription 3 (STAT3) signaling to revert E4-binding protein 4 (E4BP4)-dependent interleukin-10 (IL-10)/IL-12 imbalance and arginase I expression, thus blunting immunosuppression. Conclusions Our findings demonstrate that targeting myeloid-intrinsic CCRK signal can amplify anti-tumor T cell responses. As we also showed CCRK overexpression in patient-derived MDSCs, our results not only unravel mechanistic insights in MDSC identity but also offer a novel therapeutic kinase-target for a combinational immunotherapy strategy for conferring durable eradication of solid tumors.

Retinitis pigmentosa (RP) is a group of blinding disorders caused by diverse mutations, including in rhodopsin (RHO). Effective therapies have yet to be discovered. The I307N Rho mouse is a light-inducible model of autosomal dominant RP. Our purpose was to describe the glial response in this mouse model to educate future experimentation. I307N Rho mice were exposed to 20,000 lx of light for thirty minutes to induce retinal degeneration. Immunofluorescence staining of cross-sections and flat-mounts was performed to visualize the response of microglia and Müller glia. Histology was correlated with spectral-domain optical coherence tomography imaging (SD-OCT). Microglia dendrites extended between photoreceptors within two hours of induction, withdrew their dendrites between twelve hours and one day, appeared ameboid by three days, and assumed a ramified morphology by one month. Glial activation was more robust in the inferior retina and modulated across the boundary of light damage. SD-OCT hyper-reflectivity overlapped with activated microglia. Finally, microglia transiently adhered to the RPE before which RPE cells appeared dysmorphic. Our data demonstrate the spatial and temporal pattern of glial activation in the I307N Rho mouse, and correlate these patterns with SD-OCT images, assisting in interpretation of SD-OCT images in preclinical models and in human RP.

Background The emergence and spread of multidrug-resistant gram-negative bacteria (MDR) has become a major public health concern worldwide. This resistance is caused by enzymes-mediated genes (i.e., extended spectrum beta-lactamases) that are common in certain Enterobacterioceae species. However, the distribution of these genes is poorly documented in Burkina Faso. This study aims to determine the prevalence and distribution of the resistant genes coding for broad spectrum beta-lactamases and quinolones in rural Burkina Faso. Methods Multiplex PCR assays were carried out to detect ESBL-encoding genes, including blaOXA, blaTEM, blaCTX-M, blaSHV. The assays also assessed the presence of quinolone resistance gene namely qnrA, qnrB and qnrS in the quinolone-resistance DEC and Salmonella strains. Results The Extended-Spectrum Beta-Lactamases (ESBL) resistance phenotype was reported in all the E. coli isolates (5/5). Cross-resistance phenotype to quinolones (CRQ) was shown by one Salmonella strain (1/9) and three E. coli (3/5). Cross-resistance phenotypes to fluoroquinolones (CRFQ) were harboured by one Salmonella (1/9) and carbapenemase phenotypes were detected in two E. coli strains (2/5). Whilst the blaOXA genes were detected in 100% (5/5) of E. coli isolates and in 33.33% (3/9) Salmonella isolates. One strain of E. coli (1/5) harbored the blaCTX−M gene and the qnrB gene simultaneously. Conclusions This study identified β-lactam (bla) and quinolone resistance (qnr) genes in multidrug-resistant E. coli and Salmonella spp. in rural Burkina Faso. Our finding which highlighted the enterobacteriaceae strains resistance to β-lactams and quinolones are of high interest for adequate management of antimicrobial resistant genes outbreak in Burkina Faso.

The suppressor of cytokine signaling (SOCS) family of intracellular checkpoint inhibitors has received little recognition compared to other checkpoint inhibitors. Two members of this family, SOCS1 and SOCS3, are indispensable, since SOCS1 knockout in mice results in neonatal death due to interferon gamma (IFNI³) induced inflammatory disease, and SOCS3 knockout leads to embryonic lethality. We have shown that SOCS1 and SOCS3 (SOCS1/3) function as virus induced intrinsic virulence factors for influenza A virus, EMC virus, herpes simplex virus 1 (HSV-1), and vaccinia virus infections. Other viruses such as pathogenic pig enteric coronavirus and coronavirus induced severe acute respiratory syndrome (SARS) spike protein also induce SOCS virus intrinsic virulence factors. SOCS1/3 exert their viral virulence effect via inhibition of type I and type II interferon (IFN) function. Specifically, the SOCS bind to the activation loop of receptor-associated tyrosine kinases JAK2 and TYK2 through the SOCS kinase inhibitory region (KIR), which inhibits STAT transcription factor activation by the kinases. Activated STATs are required for IFN function. We have developed a small peptide antagonist of SOCS1/3 that blocks SOCS1/3 inhibitory activity and prevents virus pathogenesis. The antagonist, pJAK2(1001-1013), is comprised of the JAK2 activation loop, phosphorylated at tyrosine 1007 with a palmitate for cell penetration. The remarkable thing about SOCS1/3 is that it serves as a broad, simple tool of perhaps most pathogenic viruses to avoid innate host IFN defense. We suggest in this Perspective that SOCS1/3 antagonist is a simple counter measure to SOCS1/3 and should be an effective mechanism as a prophylactic and/or therapeutic against the COVID-19 pandemic that is caused by coronavirus SARS-CoV2.

Microglia are immune cells of the central nervous system capable of distinct phenotypic changes and migration in response to injury. These changes most notably include the retraction of fine dendritic structures and adoption of a globular, phagocytic morphology. Due to their characteristic responses, microglia frequently act as histological indicators of injury progression. While algorithms seeking to automate microglia counts and morphological analysis are becoming increasingly popular, few exist that are adequate for use within the retina and manual analysis remains prevalent. To address this, we propose a novel segmentation routine, implemented within FIJI-ImageJ, to perform automated segmentation and cell counting of retinal microglia. We show that our routine could perform cell counts with accuracy similar to manual observers using the I307N Rho model. Tracking cell position relative to retinal vasculature, we observed population migration towards the photoreceptor layer beginning 12 h post light damage. Using feature selection with Chi2 and principal component analysis, we resolved cells along a morphological gradient, demonstrating that extracted features were sufficiently descriptive to capture subtle morphological changes within cell populations in I307N Rho and Balb/c TLR2−/− retinal degeneration models. Taken together, we introduce a novel automated routine capable of efficient image processing and segmentation. Using data retrieved following segmentation, we perform morphological analysis simultaneously on whole populations of cells, rather than individually. Our algorithm was built entirely with open-source software, for use on retinal microglia.

National networks of laboratory-based surveillance of antimicrobial resistance (AMR) monitor resistance trends and disseminate these data to AMR stakeholders. Whole-genome sequencing (WGS) can support surveillance by pinpointing resistance mechanisms and uncovering transmission patterns. However, genomic surveillance is rare in low- and middle-income countries. Here, we implement WGS within the established Antimicrobial Resistance Surveillance Program of the Philippines via a binational collaboration. In parallel, we characterize bacterial populations of key bug-drug combinations via a retrospective sequencing survey. By linking the resistance phenotypes to genomic data, we reveal the interplay of genetic lineages (strains), AMR mechanisms, and AMR vehicles underlying the expansion of specific resistance phenotypes that coincide with the growing carbapenem resistance rates observed since 2010. Our results enhance our understanding of the drivers of carbapenem resistance in the Philippines, while also serving as the genetic background to contextualize ongoing local prospective surveillance., Whole-genome sequencing (WGS) can support drug resistance surveillance, but is rare in low- and middle-income countries. Here, the authors integrate WGS within the national antimicrobial resistance surveillance program in the Philippines and conduct a retrospective sequencing survey to characterize bacterial populations and dissect resistance phenotypes.

Avian hepatitis E virus (aHEV), clinically important in poultry industry, can cause death and reduce egg production of chickens, resulting in significant economic losses in the poultry industry. However, little is known about this aHEV infection in Burkina Faso. This study presents the results of distribution and characterization of aHEV in domestic and wild birds without clinical disease. In total 173 birds liver samples were collected from four Burkina Faso provinces, between February 2015 and June 2016. Reverse transcription polymerase chain reaction (RT-PCR) with aHEV specific degenerate primers was used to screen the presence of aHEV. RNA of aHEV was detected in 29 (16.8%) liver samples. Of these, the prevalence was diverse in different species of birds; the most frequent level was 35.3% in Numida meleagris, respectively followed by 23.5% in Gallus gallus domesticus, 13.3% in Streptopelia turtur, 13.3% in Columba livia, 6.7% in Anas platyrhynchos and 3.3% in Pternistis natalensis. The present study firstly revealed the prevalence of HEV infection in six species of birds in Burkina. It is therefore important to conduct further research on the impact on poultry mortality and egg production in our country. Key words: Avian hepatitis E virus, zoonosis, birds, prevalence, Burkina Faso.

Ovarian cancer is the most lethal cancer of the female reproductive system. In that regard, several epidemiological studies suggest that long‐term exposure to estrogen could increase ovarian cancer risk, although its precise role remains controversial. To decipher a mechanism for this, we previously generated a mathematical model of how estrogen‐mediated upregulation of the transcription factor, E2F6, upregulates the ovarian cancer stem/initiating cell marker, c‐Kit, by epigenetic silencing the tumor suppressor miR‐193a, and a competing endogenous (ceRNA) mechanism. In this study, we tested that previous mathematical model, showing that estrogen treatment of immortalized ovarian surface epithelial cells upregulated both E2F6 and c‐KIT, but downregulated miR‐193a. Luciferase assays further confirmed that microRNA‐193a targets both E2F6 and c‐Kit. Interestingly, ChIP‐PCR and bisulphite pyrosequencing showed that E2F6 also epigenetically suppresses miR‐193a, through recruitment of EZH2, and by a complex ceRNA mechanism in ovarian cancer cell lines. Importantly, cell line and animal experiments both confirmed that E2F6 promotes ovarian cancer stemness, whereas E2F6 or EZH2 depletion derepressed miR‐193a, which opposes cancer stemness, by alleviating DNA methylation and repressive chromatin. Finally, 118 ovarian cancer patients with miR‐193a promoter hypermethylation had poorer survival than those without hypermethylation. These results suggest that an estrogen‐mediated E2F6 ceRNA network epigenetically and competitively inhibits microRNA‐193a activity, promoting ovarian cancer stemness and tumorigenesis.

Tomato fruit enormously consumed in Burkina Faso is a source of health risk due to its quality. The plant of tomato cultivated in most the industrially polluted area as Ouagadougou, accumulates significant quantities of heavy metals and microorganisms and could be dangerous for consumption. The present study was conducted to investigate heavy metal and microbial contamination in soils wastewater and vegetables (tomato fruit) and to evaluate the possible health risks associated with the consumption of vegetables. Total concentrations of As, Cd, Cr, Cu, Hg, Ni, Se and Zn were determined using atomic absorption spectrophotometry. Transfer factor (TF), daily intake of metals (DIM) and health risk index (HRI) were also calculated. Microbial quality was analyzed for the presence of foodborne pathogens. The pH involved from 6.50 ± 0.1 to 9.40 ± 0.12 respectively for Boulmiougou and Kossodo. The mean metal concentrations indicated that soils were strongly polluted followed by wastewater and vegetable. The range of the mean values (in mg · kg-1) exhibited by As, Cd, Cr, Cu, Hg, Ni, Se and Zn in tomato fruit was 17.80 ± 0.32, 2.71 ± 0.02, 11.57 ± 0.2, 203.41 ± 1.20, 15.13 ± 0.30, 53.18 ± 0.20, 38.56 ± 0.10, 109.13 ± 0.25 concerning samples of Boulmiougou and 3.47 ± 0.15, 2.78 ± 0.01, 22.11 ± 0.1, 242.85 ± 2.10, 14.86 ± 0.10, 132.19 ± 1.50, 33.23 ± 0.15, 146.43 ± 1.12 for Kossodo. Microbial count was below to acceptable limits for vegetable, but underlined the presence of spoiler and foodborne pathogens. However, the daily intake of metals (DIM) and the health risk index (HRI) values were found to be under to the prescribed maximum tolerable.

Red onion bulb is a vegetable containing micronutrients with antioxidant properties. However, certain factors can have an impact on the content of these compounds. The present research evaluate the effects of parameters such as cultural practices, storage conditions and culinary practices, on the antioxidants content of red onion bulb of some varieties grown in Burkina Faso. The identification of the factors was carried out through field surveys, and their effects on the antioxidants content were assessed and appreciated through especially specific related bibliographic and laboratory test data. The results showed that84.78% of farmers use chemical fertilizers and all of them use chemical pesticides on onion crop. They don’t use premises or equipment suitable for storage. About dishes,7.61% of cooks, peel, cut and wash onion bulb before steam cooking, boiling water cooking or oil frying them at high temperature during 30 minutes to 3 hours. Some cooks use to braise onion bulb scales as ingredients for different dishes such as barbecue. Only 02,72% of cooks grind the onion bulbs with a pestle, before using the crushed as such on barbecue, or slow cooking it in different sauces. Research results show that these culinary practices contribute declining significantly the final intake of antioxidant for the consumer's body.

Background & Aims: Dysregulation of metabolism plays an important role in the development and progression of cancers, while the underlying mechanisms remain largely unknown. This study aims to explore the regulation and relevance of glycolysis in chemoresistance of gastric cancer. Methods: Biochemical differences between chemoresistant and chemosensitive cancer cells were determined by metabolism profiling, microarray gene expression, PCR or western blotting. Cancer cell growth in vitro or in vivo were analyzed by viability, apoptosis and nude mice assay. Immunoprecipation was used to explore the interaction of proteins with other proteins or DNAs. Results: By metabolic and gene expression profiling, we found that pyruvate dehydrogenase kinase 3 (PDK3) was highly expressed to promote glycolysis in chemoresistant cancer cells. Its genetic or chemical inhibition reverted chemoresistance in vitro and in vivo. It was transcriptionally regulated by transcription factor HSF1 (Heat shock factor 1). Interestingly, PDK3 can localize in the nucleus and interact with HSF1 to disrupt its phosphorylation by GSK3β. Since HSF1 was subjected to FBXW7-catalyzed polyubiquitination in a phosphorylation-dependent manner, PDK3 prevented HSF1 from proteasomal degradation. Thus, metabolic enzyme PDK3 and transcription factor HSF1 forms a positive feedback loop to promote glycolysis. As a result, inhibition of HSF1 impaired enhanced glycolysis and reverted chemoresistance both in vitro and in vivo. Conclusions: PDK3 forms a positive feedback loop with HSF1 to drive glycolysis in chemoresistance. Targeting this mitonuclear communication may represent a novel approach to overcome chemoresistance.

Rabile is a ferment of dolo, a traditional sorghum beer of Burkina Faso and it is harvested at the end of dolo fermentation process. It is a significant source of proteins and it is used like a condiment and food seasoning by some communities. The present study characterized and identified the yeasts isolated from Rabile. A total of 70 samples were collected from 7 localities of Burkina Faso with dolo sellers according to the ethnic groups. The aerobic mesophilic flora count from Rabile varied from 8.34 ± 0.72 to 10.07 ± 0.51 log10 CFU·g-1 and yeast varied from 7.24 to 8.28 log10 UFC·g-1. Based on morphological, cultural, sexual and biochemical (carbon and nitrogen assimilation) profiles, 50 yeast strains were identified and found to belong to 4 genera and 7 species. Saccharomyces cerevisiae was found as most predominant yeasts species of yeast in of Rabile with 64 %, Rhodotorula muciloginosa, (8%), Candida pseudorhangii (6%), Candida heliconiae (12%), Candida utilis (4%), Shizosaccharomyces pombe (4%) and Sporobolomyces odoratus (2%). This activity has been carried out in the Laboratory of Biotechnology for Food and Nutritional Sciences, University Ouaga I Pr Josph KY-ZERBO, Burkina Faso, and it was done from June to October 2018.

Environmental pollution can have detrimental effects on crop yield and its consumers. The current study was designed to investigate the potential human health risks associated with the consumption of lettuce crop contaminated with toxic heavy metals and microbiological status. Irrigated water, soil and lettuce were analyzed by AAS technics for heavy metals including Cd, Cr, Cu, Mn, Ni, Pb and Zn. Transfer factor (TF), daily intake of metals (DIM) and health risk index (HRI) were also calculated. Microbial analysis was carried out for the presence foodborne pathogens. The results showed that the heavy metals contents were higher in the soil than wastewater and the vegetables. Heavy metals ranged (mg·Kg-1) for Cd (1.27 to 2.93), Cr (7.28 to 7.38), Cu (0.91 to 1.70), Mn (0.29 to 6.60), Ni (1.74 to 2.16), Pb (1.32 to 1.69), Zn (3.08 to 3.79); and were higher than the WHO maximum limit permissible (ML) in vegetables. HRI < 1 indicates minimal risk. TF values designated an enhanced bio-contamination. Microbial numeration revealed the presence of spoiler and pathogenic microorganisms. The lettuce tested was not safe for human use, especially for direct consumption by human beings.

Obesity increases the risk of hepatocellular carcinoma (HCC) especially in men, but the molecular mechanism remains obscure. Here, we show that an androgen receptor (AR)-driven oncogene, cell cycle-related kinase (CCRK), collaborates with obesity-induced pro-inflammatory signaling to promote non-alcoholic steatohepatitis (NASH)-related hepatocarcinogenesis. Lentivirus-mediated Ccrk ablation in liver of male mice fed with high-fat high-carbohydrate diet abrogates not only obesity-associated lipid accumulation, glucose intolerance and insulin resistance, but also HCC development. Mechanistically, CCRK fuels a feedforward loop by inducing STAT3-AR promoter co-occupancy and transcriptional up-regulation, which in turn activates mTORC1/4E-BP1/S6K/SREBP1 cascades via GSK3β phosphorylation. Moreover, hepatic CCRK induction in transgenic mice stimulates mTORC1-dependent G−csf expression to enhance polymorphonuclear myeloid-derived suppressor cell recruitment and tumorigenicity. Finally, the STAT3-AR-CCRK-mTORC1 pathway components are concordantly over-expressed in human NASH-associated HCCs. These findings unveil the dual roles of an inflammatory-CCRK circuitry in driving metabolic and immunosuppressive reprogramming through mTORC1 activation, thereby establishing a pro-tumorigenic microenvironment for HCC development., Obesity increases the risk of hepatocellular carcinoma (HCC) especially in men. Here the authors find a potential mechanistic explanation by showing that, in mice, obesity-induced STAT3 cooperates with the androgen receptor to activate the mTORC pathway through up regulation of CCRK, resulting in hepatic steatosis worsening and HCC development via metabolic and immune reprogramming.

The microbial activity estimated by respirometry test method made it possible to measure the quantity of CO2 produced per day, from four soils polluted at different degrees by hydrocarbons. These samples have been wet to 2/3 of their maximal capacity of retention and incubated at 30°C for 30 days. The quantity of CO2 released and measured shows a high value (83.97 mg/100 g of soils against 36.05 mg/100 g of soils) in soils highly polluted by hydrocarbons treated without cow dung. We get higher values (148.98 mg/100 g of polluted soils against 66.52 mg/100 g of not contaminated soils) when the cow dung is used as a ferment. The total aerobic mesophilic flora in the samples of soils has been counted on the nutritive agar. The result is that the total aerobic mesophilic flora varied from 1×104 Colony Forming Unit (CFU) to 50 × 104 CFU before treatment and from 5×104 CFU to 420×104 CFU after 30 days of treatment. During the treatment of the soils, the bacteria and molds microflora increases with time while yeasts disappear progressively. Key words: Respirometry, microorganisms, contaminated soils, hydrocarbons.

Lung cancer is the most prevalent cancer type globally. Over 1.8 million new cases are diagnosed and 1.6 million deaths are reported yearly. The 5-year survival rates of lung cancer vary in different disease stages, histological types, genders and regions. Early screening, detection and treatment are helpful to improve survival rates (1). In the last few years, targeted therapy is becoming a promising strategy for advanced lung cancer patients.

Fibroblast growth factors (FGFs) and their receptors are significant components during fundamental cellular processes. FGF18 plays a distinctive role in modulating the activity of both tumor cells and tumor microenvironment. This study aims to comprehensively investigate the expression and functional role of FGF18 in gastric cancer (GC) and elucidate its regulatory mechanisms. The upregulation of FGF18 was detected in seven out of eleven (63.6%) GC cell lines. In primary GC samples, FGF18 was overexpressed in genomically stable and chromosomal instability subtypes of GC and its overexpression was associated with poor survival. Knocking down FGF18 inhibited tumor formation abilities, induced G1 phase cell cycle arrest and enhanced anti-cancer drug sensitivity. Expression microarray profiling revealed that silencing of FGF18 activated ATM pathway but quenched TGF-β pathway. The key factors that altered in the related signaling were validated by western blot and immunofluorescence. Meanwhile, treating GC cells with human recombinant FGF18 or FGF18-conditioned medium accelerated tumor growth through activation of ERK-MAPK signaling. FGF18 was further confirmed to be a direct target of tumor suppressor, miR-590-5p. Their expressions showed a negative correlation in primary GC samples and more importantly, re-overexpression of FGF18 partly abolished the tumor-suppressive effect of miR-590-5p. Our study not only identified that FGF18 serves as a novel prognostic marker and a therapeutic target in GC but also enriched the knowledge of FGF-FGFR signaling during gastric tumorigenesis.

Objective Resistance to antibiotics most especially third generation cephalosporins has assumed a worrisome dimension globally. Genes conferring these resistance which are mediated by enzymes known as extended spectrum beta-lactamases (ESBLs) are now wide spread among several Enterobacteriaceae species. However there is paucity of data regarding the distribution of these genes in Burkina Faso. Hence this prospective study aims to determine the prevalence and distribution of ESBL encoding genes in ESBL producing Enterobacteriaceae strains isolated from clinical samples of patients attending the three major hospitals in Ouagadougou Burkina Faso. Results ESBL-encoding genes were assayed in 187 ESBL producing Enterobacteriaceae strains. Among these isolates, the prevalence of ESBL-producing strains with blaTEM, blaSHV and blaCTX-M genes were 26.2% (49/187), 5.9% (11/187) and 40.1% (75/187) respectively. The association of ESBL encoding genes with health centers was statistically significant (p = 0.0209). Approximately 39.6% of E. coli harbored CTX-M and Klebsiella spp. 5.9%. This study demonstrates the dissemination of TEM, SHV and CTX-M genes in ESBL producing Enterobacteriaceae strains in Ouagadougou. Continuous spread of these bacteria poses great public health risk, thus increased surveillance and regulation of antibiotics use is imperative in Burkina Faso.

Members in Motin family, or Angiomotins (AMOTs), are adaptor proteins that localize in the membranous, cytoplasmic or nuclear fraction in a cell context-dependent manner. They control the bioprocesses such as migration, tight junction formation, cell polarity, and angiogenesis. Emerging evidences have demonstrated that AMOTs participate in cancer initiation and progression. Many of the previous studies have focused on the involvement of AMOTs in Hippo-YAP1 pathway. However, it has been controversial for years that AMOTs serve as either positive or negative growth regulators in different cancer types because of the various cellular origins. The molecular mechanisms of these opposite roles of AMOTs remain elusive. This review comprehensively summarized how AMOTs function physiologically and how their dysregulation promotes or inhibits tumorigenesis. Better understanding the functional roles of AMOTs in cancers may lead to an improvement of clinical interventions as well as development of novel therapeutic strategies for cancer patients.

Infectious bursal disease (IBD) is a highly contagious, immunosuppressive disease of chickens and causes substantial economic losses to the poultry industry globally. This study investigated the genetic characteristics and pathological lesions induced by IBD viruses (IBDVs) that were associated with 60 suspected outbreaks in chickens during 2015-2016 in Lusaka Province, Zambia. Nucleotide sequences of VP2 hypervariable region (VP2-HVR) (n = 38) and part of VP1 (n = 37) of Zambian IBDVs were phylogenetically analysed. Phylogenetic analysis of the VP2-HVR and VP1 revealed that most viruses (n = 31 of each genome segment) clustered with the very virulent (vv) strains. The rest of the viruses clustered with the classical strains, with two of the viruses being closely related to attenuated vaccine isolates. Two of the viruses that belonged to the vv genotype had a unique amino acid (aa) substitution Q324L whereas one virus had two unique changes, N280S and E300A in the VP2-HVR aa sequence. Although Zambian strains with a vv genotype possessed virulence marker aa within VP1 at 145T, 146D and 147N, two viruses showed unique substitutions, with one virus having 147T while the other had 147H. Pathologically, it was noted that only viruses with a vv genotype appeared to be associated with inducing pathological lesions in non-lymphoid organs (proventriculus and gizzard). Whilst documenting for the first time the presence of classical virulent IBDVs, this study demonstrates the involvement of multiple genotypes, with predominance of vvIBDVs in the epidemiology of IBD in Zambia.

We use topographic data to show that impact craters with pitted floor deposits are among the deepest on Mars. This is consistent with the interpretation of pitted materials as primary crater-fill impactite deposits emplaced during crater formation. Our database consists of 224 pitted material craters ranging in size from ∼1 to 150 km in diameter. Our measurements are based on topographic data from the Mars Orbiter Laser Altimeter (MOLA) and the High-Resolution Stereo Camera (HRSC). We have used these craters to measure the relationship between crater diameter and the initial post-formation depth. Depth was measured as maximum rim-to-floor depth, ( d r ), but we also report the depth measured using other definitions. The database was down-selected by refining or removing elevation measurements from “problematic” craters affected by processes and conditions that influenced their d r /D, such as pre-impact slopes/topography and later overprinting craters. We report a maximum (deepest) and mean scaling relationship of d r = ( 0.347 ± 0.021 ) D 0.537 ± 0.017 and d r = ( 0.323 ± 0.017 ) D 0.538 ± 0.016 , respectively. Our results suggest that significant variations between previously-reported MOLA-based d r vs. D relationships may result from the inclusion of craters that: 1) are influenced by atypical processes ( e.g. , highly oblique impact), 2) are significantly degraded, 3) reside within high-strength regions, and 4) are transitional (partially collapsed). By taking such issues into consideration and only measuring craters with primary floor materials, we present the best estimate to date of a MOLA-based relationship of d r vs. D for the least-degraded complex craters on Mars. This can be applied to crater degradation studies and provides a useful constraint for models of complex crater formation.

Slit-Robo GTPase-activating protein 1 (SRGAP1) functions as a GAP for Rho-family GTPases and downstream of Slit-Robo signaling. We aim to investigate the biological function of SRGAP1 and reveal its regulation by deregulated microRNAs (miRNAs) in gastric cancer (GC). mRNA and protein expression of SRGAP1 were examined by quantitative reverse transcription PCR (qRT-PCR) and western blot. The biological role of SRGAP1 was demonstrated through siRNA-mediated knockdown experiments. The regulation of SRGAP1 by miR-340 and miR-124 was confirmed by western blot, dual luciferase activity assays and rescue experiments. SRGAP1 is overexpressed in 9 out of 12 (75.0%) GC cell lines. In primary GC samples from TCGA cohort, SRGAP1 shows gene amplification in 5/258 (1.9%) of cases and its mRNA expression demonstrates a positive correlation with copy number gain. Knockdown of SRGAP1 in GC cells suppressed cell proliferation, reduced colony formation, and significantly inhibited cell invasion and migration. Luciferase reporter assays revealed that SRGAP1 knockdown significantly inhibited Wnt/β-catenin pathway. In addition, SRGAP1 was found to be a direct target of two tumor-suppressive miRNAs, miR-340 and miR-124. Concordantly, these two miRNAs were downregulated in primary gastric tumors and these decreasing levels w5ere associated with poor outcomes. Expression of miR-340 and SRGAP1 displayed a reverse relationship in primary samples and re-expressed SRGAP1, rescued the anti-cancer effects of miR-340. Taken together, these data strongly suggest that, apart from gene amplification and mutation, the activation of SRGAP1 in GC is partly due to the downregulation of tumor-suppressive miRNAs, miR-340 and miR-124. Thus SRGAP1 is overexpressed in gastric carcinogenesis and plays an oncogenic role through activating Wnt/β-catenin pathway.

The present study aims at optimizing the production of Saccharomyces cerevisiae SKM10 single cell proteins (SCP) from mango waste. To optimize the production process, the effect of two independent variables was evaluated by a three-level full factorial design using response surface methodology (RSM). The parameters studied were inoculum size (2 to 12% v/v) and concentration of substrates (5 to 10 g/L). Analytical methods were used to determine yeast cell biomass, sugar and protein content in yeast. Under optimal conditions, sugar content and maximum cell dry weight attained were 15.28 and 29.85% (w/w). Optimal protein content of 79.14% (w/w) was achieved at 8% (v/v) inoculum and 8% (g/L) substrate concentration. These results have provided useful information on how to improve the production by RSM and suggested that S. cerevisiae SKM10 might be applied effectively to produce SCP using mango waste as a low-cost substrate. Key words: Mango waste, Saccharomyces cerevisiae, bioconversion, production, single cell protein.

CCCTC-binding factor (CTCF) is a DNA-binding protein that interacts with a large number of highly divergent target sequences throughout the genome. It is implicated in a variety of functions, including chromatin organization and transcriptional control. The functional role of CTCF in tumour pathogenesis remains elusive. We showed that CTCF is frequently upregulated in a subset of primary hepatocellular carcinomas (HCCs) as compared with non-tumoural liver. Overexpression of CTCF was associated with shorter disease-free survival of patients. Short hairpin RNA (shRNA)-mediated suppression of CTCF inhibited cell proliferation, motility and invasiveness in HCC cell lines; these effects were correlated with prominent reductions in the expression of telomerase reverse transcriptase (TERT), the shelterin complex member telomerase repeat-binding factor 1, and forkhead box protein M1 (FOXM1). In contrast, upregulation of CTCF was positively correlated with FOXM1 and TERT expression in clinical HCC biopsies. Depletion of CTCF resulted in reduced motility and invasiveness in HCC cells that could be reversed by ectopic expression of FOXM1, suggesting that FOXM1 is one of the important downstream effectors of CTCF in HCC. Reporter gene analysis suggested that depletion of CTCF is associated with reduced FOXM1 and TERT promoter activity. Chromatin immunoprecipitation (ChIP)-polymerase chain reaction (PCR) analysis further revealed occupancy of the FOXM1 promoter by CTCF in vivo. Importantly, depletion of CTCF by shRNA significantly inhibited tumour progression and metastasis in HCC mouse models. Our work uncovered a novel functional role of CTCF in HCC pathogenesis, which suggests that targeting CTCF could be further explored as a potential therapeutic strategy for HCC. © 2017 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland.

ObjectiveMyeloid-derived suppressor cells (MDSCs) contribute to tumour immunosuppressive microenvironment and immune-checkpoint blockade resistance. Emerging evidence highlights the pivotal functions of cyclin-dependent kinases (CDKs) in tumour immunity. Here we elucidated the role of tumour-intrinsic CDK20, or cell cycle-related kinase (CCRK) on immunosuppression in hepatocellular carcinoma (HCC).DesignImmunosuppression of MDSCs derived from patients with HCC and relationship with CCRK were determined by flow cytometry, expression analyses and co-culture systems. Mechanistic studies were also conducted in liver-specific CCRK-inducible transgenic (TG) mice and Hepa1–6 orthotopic HCC models using CRISPR/Cas9-mediated Ccrk depletion and liver-targeted nanoparticles for interleukin (IL) 6 trapping. Tumorigenicity and immunophenotype were assessed on single or combined antiprogrammed death-1-ligand 1 (PD-L1) therapy.ResultsTumour-infiltrating CD11b+CD33+HLA-DR− MDSCs from patients with HCC potently inhibited autologous CD8+T cell proliferation. Concordant overexpression of CCRK and MDSC markers (CD11b/CD33) positively correlated with poorer survival rates. Hepatocellular CCRK stimulated immunosuppressive CD11b+CD33+HLA-DR− MDSC expansion from human peripheral blood mononuclear cells through upregulating IL-6. Mechanistically, CCRK activated nuclear factor-κB (NF-κB) via enhancer of zeste homolog 2 (EZH2) and facilitated NF-κB-EZH2 co-binding to IL-6 promoter. Hepatic CCRK induction in TG mice activated the EZH2/NF-κB/IL-6 cascade, leading to accumulation of polymorphonuclear (PMN) MDSCs with potent T cell suppressive activity. In contrast, inhibiting tumorous Ccrk or hepatic IL-6 increased interferon γ+tumour necrosis factor-α+CD8+ T cell infiltration and impaired tumorigenicity, which was rescued by restoring PMN-MDSCs. Notably, tumorous Ccrk depletion upregulated PD-L1 expression and increased intratumorous CD8+ T cells, thus enhancing PD-L1 blockade efficacy to eradicate HCC.ConclusionOur results delineate an immunosuppressive mechanism of the hepatoma-intrinsic CCRK signalling and highlight an overexpressed kinase target whose inhibition might empower HCC immunotherapy.

We propose a new method for determining the target genes of transcriptional enhancers in specific cells and tissues. It combines global trends across many samples and sample-specific information, and considers the joint effect of multiple enhancers. Our method outperforms existing methods when predicting the target genes of enhancers in unseen samples, as evaluated by independent experimental data. Requiring few types of input data, we are able to apply our method to reconstruct the enhancer-target networks in 935 samples of human primary cells, tissues and cell lines, which constitute by far the largest set of enhancer-target networks. The similarity of these networks from different samples closely follows their cell and tissue lineages. We discover three major co-regulation modes of enhancers and find defense-related genes often simultaneously regulated by multiple enhancers bound by different transcription factors. We also identify differentially methylated enhancers in hepatocellular carcinoma (HCC) and experimentally confirm their altered regulation of HCC-related genes.

Background A genomic signal track is a set of genomic intervals associated with values of various types, such as measurements from high-throughput experiments. Analysis of signal tracks requires complex computational methods, which often make the analysts focus too much on the detailed computational steps rather than on their biological questions. Results Here we propose Signal Track Query Language (STQL) for simple analysis of signal tracks. It is a Structured Query Language (SQL)-like declarative language, which means one only specifies what computations need to be done but not how these computations are to be carried out. STQL provides a rich set of constructs for manipulating genomic intervals and their values. To run STQL queries, we have developed the Signal Track Analytical Research Tool (START, http://yiplab.cse.cuhk.edu.hk/start/), a system that includes a Web-based user interface and a back-end execution system. The user interface helps users select data from our database of around 10,000 commonly-used public signal tracks, manage their own tracks, and construct, store and share STQL queries. The back-end system automatically translates STQL queries into optimized low-level programs and runs them on a computer cluster in parallel. We use STQL to perform 14 representative analytical tasks. By repeating these analyses using bedtools, Galaxy and custom Python scripts, we show that the STQL solution is usually the simplest, and the parallel execution achieves significant speed-up with large data files. Finally, we describe how a biologist with minimal formal training in computer programming self-learned STQL to analyze DNA methylation data we produced from 60 pairs of hepatocellular carcinoma (HCC) samples. Conclusions Overall, STQL and START provide a generic way for analyzing a large number of genomic signal tracks in parallel easily. Electronic supplementary material The online version of this article (doi:10.1186/s12864-017-4071-1) contains supplementary material, which is available to authorized users.

Diarrheagenic Escherichia coli (DEC) is important bacteria of children's endemic and epidemic diarrhea worldwide. The aim of this study was to determine the prevalence of DEC isolated from stool samples collected from children with acute diarrhea living in Ouagadougou, Burkina Faso. From August 2013 to October 2015, stool samples were collected from 315 children under 5 years of age suffering from diarrhea in the "Centre Medical avec Antenne Chirurgicale (CMA)" Paul VI and the CMA of Schiphra. E. coli were isolated and identified by standard microbiological methods, and the 16-plex PCR method was used to further characterize them. Four hundred and nineteen (419) E. coli strains were characterized, of which 31 (7.4%) DEC pathotypes were identified and classified in five E. coli pathotypes: 15 enteroaggregative E. coli (EAEC) (48.4%), 8 enteropathogenic E. coli (EPEC) (25.8%) with 4 typical EPEC and 4 atypical EPEC, 4 enteroinvasive E. coli (EIEC) (12.9%), 3 enterohemorrhagic E. coli (EHEC) 9.67%, and 1 enterotoxigenic E. coli (ETEC) 3.2%. The use of multiplex PCR as a routine in clinical laboratory for the detection of DEC would be a useful mean for a rapid management of an acute diarrhea in children.

An association between periodontitis and nonalcoholic fatty liver disease (NAFLD) has been reported by experimental animal and epidemiologic studies. This study investigated whether circulating levels of serum C-reactive protein (CRP) and a weighted genetic CRP score representing markers of inflammatory burden modify the association between periodontitis and NAFLD. Data came from 2,481 participants of the Study of Health in Pomerania who attended baseline examination that occurred between 1997 and 2001. Periodontitis was defined as the percentage of sites (0%, 3 mg/L. Periodontitis was positively associated with higher prevalence odds of NAFLD, and this relationship was modified by serum CRP levels.

TEA domain (TEAD) transcription factors are key components of the Hippo-YAP1 signaling pathway, but their functional role and regulatory mechanisms remain unclear. This study aims to comprehensively explore the expression pattern and functional role of TEAD family in gastric carcinogenesis and investigate its regulation by microRNAs (miRNAs). The mRNA and protein expression of TEAD family were examined by quantitative reverse transcription-PCR (qRT-PCR) and western blot. Their functional roles were determined by in vitro and in vivo studies. The clinicopathological association of TEAD4 in gastric cancer (GC) was studied using immunohistochemistry on tissue microarray. The prediction of miRNAs, which potentially target TEAD1/4, was performed by TargetScan and miRDB. The regulation of TEAD1/4 by miRNAs was confirmed by qRT-PCR, western blot and luciferase assays. TEAD1/4 were overexpressed in GC cell lines and primary GC tissues. Knockdown of TEAD1/4 induced a significant anticancer effect in vitro and in vivo. TEAD1 was confirmed to be a direct target of miR-377-3p and miR-4269, while TEAD4 was negatively regulated by miR-1343-3p and miR-4269. Among them, miR-4269 was the most effective inhibitor of TEAD1/4. Ectopic expression of these miRNAs substantiated their tumor-suppressive effects. In primary GC tumors, downregulation of miR-4269 was associated with poor disease-specific survival and showed a negative correlation with TEAD4. TEAD1 and TEAD4 are oncogenic factors, whose aberrant activation are, in part, mediated by the silence of miR-377-3p, miR-1343-3p and miR-4269. For the first time, the nuclear accumulated TEAD4 and downregulated miR-4269 are proposed to serve as novel prognostic biomarkers in GC.

Extended spectrum beta lactamase (ESBL) producing Enterobacteriaceae is one of the main causes of antibiotic treatment failure in hospitals. The aim of this study was to evaluate the prevalence of ESBL produced by multidrug-resistant (MDR) Escherichia coli isolated from various clinical samples (urine, stool, pus, blood culture) in the “Hopital National de Niamey” and the “Hopital National Lamorde” of Niamey, Niger. Samples were processed using standard bacteriological methods. Isolates were identified by biochemical tests and confirmed on API 20 E system (Bio-Merieux, France). Antibiotic susceptibility was determined using the disk diffusion method on Mueller-Hinton (MH) agar plates (Liofilchem, Italy). Producing of extended spectrum beta-lactamase was performed using simple double-disk synergy test (DDST) and double-disk synergy test using cloxacillin. A total of two hundred and seventeen (217) multidrug-resistant E. coli were isolated from various clinical samples. Among these isolates, 57 (26.3%) were extended spectrum beta-lactamase producers. From clinical sources, prevalence of ESBL producing E. coli was observed in urine samples (26.7%), stool samples (26.3%), pus samples (25%) and blood samples (25%). ESBL producing E. coli were observed in the age groups under 5 years (24.9%), 26 to 45 (38.1%) and over 65 years (50%). This study showed a notable prevalence of extended spectrum beta-lactamase E. coli isolated from various clinical samples in two hospitals of Niamey, suggesting the rational and judicious use of antibiotics by clinicians. Key words: Extended spectrum beta lactamase (ESBL), multidrug-resistance, Escherichia coli, prevalence, Niamey, Niger.