Remarkable genetic diversity detected at river buffalo prolactin receptor (PRLR) gene and association studies with milk fatty acid composition

Prolactin is an anterior pituitary peptide hormone involved in many different endocrine activities and is essential for reproductive performance. This action is mediated by its receptor, the prolactin receptor, encoded by the PRLR gene. In this study, we sequenced and characterized the Mediterranean river buffalo PRLR gene (from exon 3 to 10), and we found remarkable genetic diversity. In particular, we found 24 intronic polymorphisms and 13 exonic SNPs, seven of which were non‐synonymous. Furthermore, the polymorphisms identified in the 3′‐UTR were investigated to establish their possible influence on microRNA binding sites. Considering all the amino acid changes and the observed allelic combinations, it is possible to deduce at least six different translations of the buffalo prolactin receptor and, consequently, the presence at the PRLR gene of at least six alleles. Furthermore, we identified a deletion of a CACTACC heptamer between nucleotides 1102 and 1103 of exon 10 (3′‐UTR), and we developed an allele‐specific PCR to identify the carriers of this genetic marker. Finally, the SNP g.11188A>G, detected in exon 10 and responsible for the amino acid replacement p.His328Arg, was genotyped in 308 Italian Mediterranean river buffaloes, and an association study with milk fat traits was carried out. The statistical analysis showed a tendency that approached significance for the AA genotype with higher contents of odd branched‐chain fatty acids. Thus, our results suggest that the PRLR gene is a good candidate for gene association studies with qualitative traits related to buffalo milk production.     

Comprehensive DNA microarray expression profiles of tumors in tenascin-C-knockout mice

Tenascin-C (TNC), an extracellular matrix glycoprotein, plays a pivotal role in tumor growth. However, the mechanism whereby TNC affects tumor biology remains unclear. To investigate the exact role of TNC in primary tumor growth, a mouse mammary tumor cell line, GLMT1, was first developed. Subsequently, global gene expression in GLMT1-derived tumors was compared between wild-type (WT) and TNC-knockout (TNKO) mice. Tumors in WT mice were significantly larger than those in TNKO mice. DNA microarray analysis revealed 447 up and 667 downregulated in the tumors inoculated into TNKO mice as compared to tumors in WT mice. Validation by quantitative gene expression analysis showed that Tnc, Cxcl1, Cxcl2, and Cxcr2 were significantly upregulated in WT mice. We hypothesize that TNC stimulates the CXCL1/2-CXCR2 pathway involved in cancer cell proliferation.     

Inhibition of acid production in Streptococcus mutans R9: Inhibition constants and reversibility

Abstract The pac gene encoding the penicillin G acylase (PGA) of Bacillus megaterium ATCC 14945 has been cloned in Escherichia coli HB101 ( proA, leuB ) using a selective minimal medium containing phenylacetyl-L-leucine instead of L-leucine. The nucleotide sequence of this gene has been determined and contains an open reading frame of 2406 nucleotides. The deduced amino acid sequence shows significant similarity with other β-lactam acylases. Although the PGA of B. megaterium is extracellular, the enzyme produced in E. coli appears to have a cytoplasmic localization.     

The Immunocytochemical Detection of P53 Protein In Cytological Specimens: Technical Considerations

The p53 gene encodes a 393 amino acid nuclear phosphoprotein that appears to act as a cell cycle checkpoint, possibly by transactivating other target genes. Abnormalities of the p53 gene are common in a wide range of human tumours and are associated in many cases with immunologically detectable p53 protein. Detection of p53 immunoreactivity is uncommon in normal cells, but is frequently seen in neoplasia. Here we define the optimum conditions for the detection of p53 immunoreactivity in cytological material, including fixation and storage. Immersion in acetone-methanol for 10 min is optimal, and after air drying, smears or cytospin preparations can be stored at - 70°C for at least 6 months. We describe the range of controls necessary, including the use of positive control cell lines with known mutations of the p53 gene and defined abnormalities of p53 protein. Negative controls should include cell lines (or strains) with no p53 abnormality as well as the conventional negative immunological controls. It is only with these technical caveats and controls that p53 immunoreactivity can be performed reliably on cytological specimens. Le géne p53 code pour une phosphoprotéine nucléaire de 393 acides aminés qui semble jouer en rôle dans la régulation du cycle cellulaire, probablement par transactivation d'autres gènes cibles. Les anomalies du gène p53 sont présentes dans un large éventail de tumeurs humaines et sont associées a la présence d'une protéine p53 détectable immunologiquement. La détection d'une immunoréactivité anti p53 est rare dans les cellules normales alors qu'elle est fréquente dans les tumeurs. Nous avons défini dans ce travail les conditions optimales pour la détection de l'immunoréactivité anti p53 sur matériel cytologique, y compris les conditions de fixation et de conservation. L'immersion dans l'acétone-méthanol pendant 10 minutes est optimale. Aprés séchage à l'air, les frottis ou les préparations par cytocentrifugation peuvent être stockés à—70°C pendant au moins 6 mois. Nous décrivons aussi l'éventail des contrôles nécessaires incluant I'utilisation, comme contrôle positif, de lignées cellulaires avec des mutations connues du gène p53 et des anomalies définies de la protéine p53. Les contrôles négatifs doivent comporter des lignées cellulaires (ou des espèces) sans anomalie de p53 ainsi que les contrôles immunologiques négatifs convrentionnels. C'est seulement lorsque ces précautions techniques et ces contrôles sont respectés que l'immunoréactivité anti p53 peut être étudiée valablement sur les prélèvements cytologiques. Das p53 kodiert ein aus 393 Aminosären bestehendes Phosphoprotein, das offensichtlich den Zellzyklus blockiert, möglicherweise durch Aktivierung anderer Gene. Veränderungen des p53-Gens wurden in zahlreichen menschlischen Tumoren nachgewiesen, sodaß eine positive Reaktion in Neoplasien häufig, in Normalzellen jedoch ungewöhnlich ist. Die optimalen Bedingungen für den p53-Nachweis in cytologischem Material werden hinsichtlich Fixation und Lagerung untersucht. Eine 10minütige Aceton-Methanol-Fixierung mit anschlies-sender Lufttrocknung erlaubt die Lagerung von Ausstrichen und Cytozentrifugen-präparaten bei - 70°C für mindestens 6 Monate. Die erforderlichen Kontrollen einschließlich positiver Zellinien mit bekannten Mutationen des p53-Gens und definierten Anomalien des Proteins werden beschrieben. Negativkontrollen sollten Zellinien ohne p53-Anomalie ebenso umfassen wie die üblichen negativen immunologischen Kontrollen. Nur unter diesen Bedingungen ist ein zuverlässiger Nachweis von p53 mögloch.     

Epigenetic Regulator CoREST Controls Social Behavior in Ants

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Screening of significant biomarkers related with prognosis of liver cancer by lncRNA-associated ceRNAs analysis

This study aimed to identify significant biomarkers related to the prognosis of liver cancer using long noncoding RNA (lncRNA)-associated competing endogenous RNAs (ceRNAs) analysis. Differentially expressed mRNA and lncRNAs between liver cancer and paracancerous tissues were screened, and the functions of these mRNAs were predicted by gene ontology and pathway enrichment analyses. A ceRNA network consisting of differentially expressed mRNAs and lncRNAs was constructed. LncRNA FENDRR and lncRNA HAND2-AS1 were hub nodes in the ceRNA network. A risk score assessment model consisting of eight genes (PDE2A, ESR1, FBLN5, ALDH8A1, AKR1D1, EHHADH, ADRA1A, and GNE) associated with prognosis were developed. Multivariate Cox regression suggested that both pathologic_T and risk group could be regarded as independent prognostic factors. Furthermore, a nomogram model consisting of pathologic_T and risk group showed a good prediction ability for predicting the survival rate of liver cancer patients. The nomogram model consisting of pathologic_T and a risk score assessment model could be regarded as an independent factor for predicting prognosis of liver cancer.     

Metabolic Reconfiguration in C. elegans Suggests a Pathway for Widespread Sterol Auxotrophy in the Animal Kingdom

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