p38alpha MAP kinase is activated in response to many cellular stresses and also regulates the differentiation and/or survival of various cell types in vitro, including skeletal muscle cells and cardiomyocytes. Here we show that targeted inactivation of the mouse p38alpha gene results in embryonic lethality at midgestation correlating with a massive reduction of the myocardium and malformation of blood vessels in the head region. However, this defect appears to be secondary to insufficient oxygen and nutrient transfer across the placenta. When the placental defect was rescued, p38alpha(-/-) embryos developed to term and were normal in appearance. Our results indicate that p38alpha is required for placental organogenesis but is not essential for other aspects of mammalian embryonic development.
Obesity in childhood and adolescence is considered the most prevalent nutritional disorder, in which eating behaviours represent one important factors of influence. Many aspects influence eating behaviours, but taste is considered the main predictor. However, data concerning correlations of obesity, taste sensitivity and behavioural attitudes, such as food neophobia, in children and adolescents are inconsistent. Moreover, it has been suggested that oral bacteria could have a possible role in obesity development and, also, in taste perception. In this context, the present study focused on host related factors with a proposed link to weight gain. To this purpose, taste sensitivity, salivary microbiota composition and food neophobia were compared between children and adolescents with and without obesity in a cross-sectional study. Results showed that children with obesity presented a significantly lower ability in correctly identifying taste qualities and were characterized by a lesser number of Fungiform Papillae (reported as FP/cm2) compared to normal-weight subjects. Differences in the ecological indexes of microbial alpha-diversity was found between subjects with obesity and normal-weight ones. Moreover, independently from nutritional status, some bacterial genera seemed to differ between subjects with different sensitivity. The potentiality of this multidisciplinary approach could help to better understand and deepen the sensory-driven and microbiological factors related to weight gain.
in this work we present easyprimer, a user-friendly online tool developed to assist pan-pcR and High Resolution Melting (HRM) primer design. The tool finds the most suitable regions for primer design in a gene alignment and returns a clear graphical representation of their positions on the consensus sequence. EasyPrimer is particularly useful in difficult contexts, e.g. on gene alignments of hundreds of sequences and/or on highly variable genes. HRM analysis is an emerging method for fast and cost saving bacterial typing and an HRM scheme of six primer pairs on five Multi-Locus Sequence Type (MLST) genes is already available for Klebsiella pneumoniae. We validated the tool designing a scheme of two HRM primer pairs on the hypervariable gene wzi of Klebsiella pneumoniae and compared the two schemes. the wzi scheme resulted to have a discriminatory power comparable to the HRM MLST scheme, using only one third of primer pairs. then we successfully used the wzi HRM primer scheme to reconstruct a Klebsiella pneumoniae nosocomial outbreak in few hours. the use of hypervariable genes reduces the number of HRM primer pairs required for bacterial typing allowing to perform cost saving, large-scale surveillance programs.Most methods used for the identification and typing of prokaryotes are based on DNA amplification and sequencing. Indeed, the sequence of specific genes can harbour enough information to classify bacteria at species, subspecies or also to a clonal level. For instance, Multi-Locus Sequence Typing (MLST) is one of the most used methods for bacterial typing and it is based on the amplification and sequencing of few housekeeping genes 1 . During the last ten years, the analysis of the entire bacterial genome by Whole Genome Sequencing (WGS) approach revolutionized the field, drastically increasing the typing precision 1 .The reconstruction of nosocomial outbreaks is one of the most important clinical applications of bacterial typing. A nosocomial outbreak occurs when the number of patients infected by a pathogen increases above the expected in a limited time 2 . In these situations, it is fundamental to determine the clonality of bacteria causing disease in the patients to define the proper strategy to handle the emergency. Pulsed-Field Gel Electrophoresis (PFGE), MLST and WGS are the most frequently applied molecular methods in outbreak investigation 1 .During a nosocomial outbreak, clinicians need bacterial typing information in the shortest time possible. Despite the high potential of WGS in outbreak reconstruction, the sequencing of a complete genome requires two to four working days, introducing an important time lag. Similarly, PFGE typing requires five days and also MLST needs few days. During the last years, the High Resolution Melting (HRM) assay has emerged as a low-cost and fast method for bacterial typing, particularly promising for epidemiological applications 3-6 . HRM is a single-step
Background: Growing evidence suggests that an altered microbiota composition contributes to the pathogenesis and clinical features in celiac disease (CD). We performed a comparative analysis of the gut microbiota in adulthood CD to evaluate whether: (i) dysbiosis anticipates mucosal lesions, (ii) gluten-free diet restores eubiosis, (iii) refractory CD has a peculiar microbial signature, and (iv) salivary and fecal communities overlap the mucosal one. Methods: This is a cross-sectional study where a total of 52 CD patients, including 13 active CD, 29 treated CD, 4 refractory CD, and 6 potential CD, were enrolled in a tertiary center together with 31 controls. A 16S rRNA-based amplicon metagenomics approach was applied to determine the microbiota structure and composition of salivary, duodenal mucosa, and stool samples, followed by appropriate bioinformatic analyses. Results: A reduction of both α- and β-diversity in CD, already evident in the potential form and achieving nadir in refractory CD, was evident. Taxonomically, mucosa displayed a significant abundance of Proteobacteria and an expansion of Neisseria, especially in active patients, while treated celiacs showed an intermediate profile between active disease and controls. The saliva community mirrored the mucosal one better than stool. Conclusion: Expansion of pathobiontic species anticipates villous atrophy and achieves the maximal divergence from controls in refractory CD. Gluten-free diet results in incomplete recovery. The overlapping results between mucosal and salivary samples indicate the use of saliva as a diagnostic fluid.
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