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International audience; To investigate if the artificial delivery of microRNAs naturally present in the breastmilk can impact the gut and brain of young rats according to weaning. Animals from a new transgenic rat line expressing the green-fluorescent protein in the endocrine lineage (cholecystokinin expressing cells) received a single oral bolus of miR-320-3p or miR-375-3p embedded in DiOleyl-Succinyl-Paromomycin (DOSP) on D-12. The pups were weaned early (D-15), or regularly (D-30). The expression of relevant miRNA, mRNAs, chromatin complexes, and duodenal cell density were assessed at 8 h post-inoculation and on D-45. The miR-320-3p/DOSP induced immediate effects on H3K4me3 chromatin complexes with polr3d promoter (p < 0.05). On regular weaning, on D-45, miR-320-3p and 375-3p were found to be downregulated in the stomach and upregulated in the hypothalamus (p < 0.001), whereas miR-320-3p was upregulated in the duodenum. After early weaning, miR-320-3p and miR-375-3p were downregulated in the stomach and the duodenum, but upregulated in the hypothalamus and the hippocampus. Combination of miR-320-3p/DOSP with early weaning enhanced miR-320-3p and chromogranin A expression in the duodenum. In the female brain stem, miR-320-3p, miR-504, and miR-16-5p levels were all upregulated. Investigating the oral miRNA-320-3p loads in the duodenal cell lineage paved the way for designing new therapeutics to avoid unexpected long-term impacts on the brain.

International audience; Apolipoproteins govern lipoprotein metabolism and are promising biomarkers of metabolic and cardiovascular diseases. Unlike immunoassays, MS enables the quantification and phenotyping of multiple apolipoproteins. Hence, here, we aimed to develop a LC-MS/MS assay that can simultaneously quantitate 18 human apolipoproteins [A-I, A-II, A-IV, A-V, B48, B100, C-I, C-II, C-III, C-IV, D, E, F, H, J, L1, M, and (a)] and determined apoE, apoL1, and apo(a) phenotypes in human plasma and serum samples. The plasma and serum apolipoproteins were trypsin digested through an optimized procedure and peptides were extracted and analyzed by LC-MS/MS. The method was validated according to standard guidelines in samples spiked with known peptide amounts. The LC-MS/MS results were compared with those obtained with other techniques, and reproducibility, dilution effects, and stabilities were also assessed. Peptide markers were successfully selected for targeted apolipoprotein quantification and phenotyping. After optimization, the assay was validated for linearity, lower limits of quantification, accuracy (biases: -14.8% to 12.1%), intra-assay variability [coefficients of variation (CVs): 1.5-14.2%], and inter-assay repeatability (CVs: 4.1-14.3%). Bland-Altman plots indicated no major statistically significant differences between LC-MS/MS and other techniques. The LC-MS/MS results were reproducible over five repeated experiments (CVs: 1.8-13.7%), and we identified marked differences among the plasma and serum samples. The LC-MS/MS assay developed here is rapid, requires only small sampling volumes, and incurs reasonable costs, thus making it amenable for a wide range of studies of apolipoprotein metabolism. We also highlight how this assay can be implemented in laboratories.

AimTo investigate if the artificial delivery of microRNAs naturally present in the breastmilk can impact the gut and brain of young rats according to weaning.MethodsAnimals from a new transgenic rat line expressing green-fluorescent protein in the endocrine lineage (cholecystokinin expressing cells) received at Day-12, near neural diversification, a single oral bolus of mir-320-3p or miR-375-3p, embedded in DiOleyl-Succinyl-Paromomycin (DOSP), and were further early (Day-15) or regularly (Day-30) weaned. Relevant miRNA (miR-320-3p, miR-375-3p, miR-375-5p, miR-16-5p, miR-132-3p, miR-504), polr3d, hspb6, inflammation, enteroendocrine, and circadian clock-related mRNAs, chromatin complexes, and duodenal cell density were assayed at 8h post-inoculation and at Day-45.ResultsThe miR-320-3p/DOSP induced immediate effects on H3K4me3 chromatin complexes with polr3d promoter (pConclusionAddressing oral miRNA-320-3p loads to duodenal cell lineage is paving the way for the design of new therapeutics, manipulating long term consequences of early life stress.

En la presente investigación el objetivo fue determinar los factores que atraen y retienen las actividades económicas del turismo, para realizar propuestas que coadyuven el desarrollo de este sector, para ello se realizó una encuesta en la que participaron 450 administradores de hoteles, restaurantes, transporte turístico y servidores públicos del área de turismo y líderes de seis comunidades del estado de Tabasco, México, los cuales respondieron de forma voluntaria al instrumento de investigación, se realizó el análisis factorial exploratorio de los datos, en la prueba piloto y en la muestra. En ambos casos se obtuvieron valores superiores a .80 en el coeficiente de fiabilidad alfa de Cronbach, así mísmo se evidenció validez en la estructura mediante la fiabilidad compuesta (CR, u Omega de Mcdonald). El análisis de los datos se realizó mediante modelos de ecuaciones estructurales (SEM), así también por medio de la estadística descriptiva y análisis de varianza ANOVA de un factor. Los resultados presentan ajuste idóneos para el modelo de investigación propuesto, se validaron como factores de atracción y retención del turismo a los recursos turísticos, infraestructura, accesibilidad, políticas públicas y la comunidad, se encontró que los stakeholders moderan los elementos del turismo y de acuerdo al modelo estructural de relaciones y a los efectos totales, directos y significativos, las variables validadas se influyen recíprocamente, por lo que deben ser consideradas de manera integral en la planificación turística. 192

International audience

International audience

Turnera diffusa (damiana) grows wild in Baja California Sur (BCS), Mexico. It is traditionally used for its aphrodisiac effects and for diabetes treatment. Two varieties (var.) have been identified: diffusa and aphro-disiaca. To evaluate the effects of water-ethanol extracts from both varieties, measurements on blood glucose, body weight, biochemical parameters, and cell blood counts were performed in a diabetic rat model. Oxidative stress was also evaluated when var. diffusa was administered. No significant reductions in blood glucose levels, in biochemical parameters, or in cell blood counts with any variety of damiana were observed; however, it helped diabetic animals to keep their body weight. Damiana var. diffusa did not mo-dify the mitochondrial nitric oxide levels and superoxide dismutase activity; however, thiobarbituric acid reactive species diminished. In conclusion, damiana can be utilized reliably in the management of diabetes because of its antioxidant potential; it promoted body weight loss in healthy animals and helped to preserve it in diabetes

The family of enzymes 5-enolpiruvil shikimato-3-phosphate synthase (EPSPS) is found in plants and microorganisms. The substrates of this enzyme are phosphoenolpyruvate (PEP) and 3-phospho-shikimate and their products are phosphate and 5-enolpyruvylshikimate-3-phosphate that is the biological target of the herbicide glyphosate, which is used in genetically modified crops. The interaction between cultivated genetically modified plants (GMP) and wild plant species could be a transference source of transgenes. Presence of transgenes could be cause and adverse environmental impact on non-target organisms. Gossypium hirsutum genotype Bollgard II® is a GMP with tolerance to herbicide glyphosate and it has been cultivated during 20 years in Mexico and the possibility to gene flow primary in congeners of the Malvaceae family is possible. The objective of this study was to quantify and identify weed species associated to genetically modified cotton fields and to detect the present of glyphosate-insensitive EPSP synthases (CP4 EPSPS) in these species. The results showed that plants of the families Amaranthaceae, Asteraceae, Boraginaceae, Chenopodiaceae, Convolvulaceae, Fabaceae, Malvaceae, Poaceae, Portulacaceae, Solanaceae and Zygophyllaceae are present in the study site. Twenty-five weed species belonging to these botanical families were collected and identified in the site. From these, two species of the Malvaceae family with potential risk of gene flow plants, Anoda cristata and Sida hederacea were identified in the site; however, the CP4 EPSPS protein was not detected in none of the collected weed species and only the GM genotype Bollgard II® was positive to the CP4 EPSPS protein in the study site.

Ovarian fibrosarcomas are extremely rare tumors with little genomic information available to date. In the present report we present the tumoral exome and transcriptome and the germinal exome of an ovarian fibrosarcoma from a 9-years old child. We found a paucity of mutations (0.77/Mb) and CNV alterations. Of these, the most relevant were a point mutation in the metal-binding site of the microRNA-processing DICER1 enzyme and a frame-shift alteration in the tumor suppressor gene NF1. We validated a germinal truncating mutation in DICER1, which was consistent with a DICER1 Syndrome diagnosis, providing the first example of an ovarian fibrosarcoma as the presenting neoplasia in this syndrome. Network and enrichment analyses showed that both a mesenchymal signature and a Hedgehog cascade could be driving the progression of this tumor. We were also able to find a global lincRNA deregulation, as the number of lincRNAs transcripts expressed in the tumor was decreased, with a concomitant upregulation of previously described non-coding transcripts associated with cancer, such as MALAT1, MIR181A1HG, CASC1, XIST and FENDRR. DICER1 Syndrome should be considered as a possible diagnosis in children ovarian fibrosarcoma. The role of lncRNAs in neoplasias associated with DICER1 alterations need to be studied in more detail.

RNA-Sequencing and de novo assembly have enabled the analysis of species with non-available reference transcriptomes, although intrinsic features (biological and technical) induce errors in the reconstruction. A strategy to resolve these errors consists of varying assembling process parameters to generate multiple reconstructions. However, the best assembly selection remains a challenge. Quantitative metrics for quality assessment have been inconsistent when compared with pertinent references. In this paper, a criterion for supporting assembly selection based on mapping DNA microarray hybridized probes to assembly sets is proposed. Mouse and fruit fly RNA-Seq datasets were assembled with standard de novo procedures. Quality assessment was estimated using quantitative metrics and the proposed criterion. The assembly that best mapped to the available reference transcriptomes of these model species provided the highest quality assembly. The hybridized probes identified the best assemblies, whereas quantitative metrics remained inconsistent. For example, subtle probe mapping difference of 0.25 percent, but statistically significant (ANOVA, p0.05), enabled the assembly selection that led to identify 3,719 more contigs and led to 1,049 further mapped contigs to the mouse reference transcriptome. The microarray data availability for non-model species makes the proposed criterion suitable for quality assessment of multiple de novo assembly strategies.

"Una fracción del extracto acuoso del alga marina Hypnea cervicornis se sometió a evaluación contra el virus del dengue, DENV-2. La fracción obtenida por precipitación con sulfato de amonio mostró en un inmunoensayo reducción del 43 % de la proteína viral no estructural NS1 en el sobrenadante de células Huh-7 infectadas con el virus. Esa observación sugiere una inhibición del proceso de infección. La actividad se confirmó con experimentos de q-PCR observándose una reducción significativa del número de copias del genoma viral a nivel intracelular (1.17 log) con respecto al control positivo de infección. Adicionalmente, la fracción evaluada mostró altos títulos de hemaglutinación y un patrón de afinidad que sugiere que la actividad antiviral observada podría ser debida a lectinas."

Pancreatic cancer is one of the most aggressive human malignancies with an increasing incidence worldwide. In addition to the poor survival rates, combinations using gemcitabine as a backbone have failed to show any benefit beyond monotherapy. These facts underscore an urgent need for novel therapeutic options and motivated us to study the effect of berberine on pancreatic cancer cells. Here, we undertook an mRNA-based gene expression profiling study in order to get deeper insight into the molecular targets mediating the growth inhibitory effects of berberine on pancreatic cancer cells compared to normal ones. Twenty-four hours after treatment, berberine showed preferential selectivity toward pancreatic cancer cells compared to normal ones. Moreover, expression profiling and Ingenuity pathway analysis results showed that the cytotoxicity of berberine was accompanied with an activation of BRCA1-mediated DNA damage response, G1/S and G2/M cell cycle checkpoint regulation, and P53 signalling pathways. The activation of these signalling pathways might be explained by the fact that berberine intercalates DNA and induces DNA strand break through inhibition of topoisomerases and induction of DNA lesions.

We describe the methodology used to analyze multiple transcripts using microarray techniques in simultaneous biopsies of muscle, adipose tissue and lymphocytes obtained from the same individual as part of the standard protocol of the Genetics of Metabolic Diseases in Mexico: GEMM Family Study.We recruited 4 healthy male subjects with BM1 20-41, who signed an informed consent letter. Subjects participated in a clinical examination that included anthropometric and body composition measurements, muscle biopsies (vastus lateralis) subcutaneous fat biopsies anda blood draw. All samples provided sufficient amplified RNA for microarray analysis. Total RNA was extracted from the biopsy samples and amplified for analysis.Of the 48,687 transcript targets queried, 39.4% were detectable in a least one of the studied tissues. Leptin was not detectable in lymphocytes, weakly expressed in muscle, but overexpressed and highly correlated with BMI in subcutaneous fat. Another example was GLUT4, which was detectable only in muscle and not correlated with BMI. Expression level concordance was 0.7 (p0.001) for the three tissues studied.We demonstrated the feasibility of carrying out simultaneous analysis of gene expression in multiple tissues, concordance of genetic expression in different tissues, and obtained confidence that this method corroborates the expected biological relationships among LEPand GLUT4. TheGEMM study will provide a broad and valuable overview on metabolic diseases, including obesity and type 2 diabetes.