Role of Gut Microbiota in Overweight Susceptibility in an Adult Population in Italy.

Although the gut microbiota is known to affect body weight, its relationship with overweight/obesity is unclear. Our aim was to characterize microbiota composition in a cohort from the southernmost area of Italy. We investigated whether an altered gut microbiota could play an etiological role in the pathogenesis of overweight/obesity. A total of 163 healthy adults were enrolled. Microbiome analysis was performed via 16S rRNA gene sequencing. We found significant phylum variations between overweight (N = 88) and normal-weight (N = 75) subjects. Bacteroidetes and Proteobacteria were higher in overweight participants (p = 0.004; p = 0.03), and Firmicutes and Verrucomicrobia were lower (p = 0.02; p = 0.008) compared to normal-weight participants. Additionally, Akkermansia and Bifidobacterium (genus level) were significantly lower in the overweight group, as well as Akkermansia muciniphila at the species level. The Firmicutes/Bacteroidetes ratio (F/B ratio), an index of dysbiosis, was found to be inversely associated with BMI in linear and logistic regression models (p = 0.001; p = 0.005). The association remained statistically significant after adjustment for potential confounders. This cross-sectional study contributes to defining the gut microbiota composition in an adult population living in southern Italy. It confirms the relationship between overweight susceptibility and the dysbiosis status, highlighting the possible etiological role of the F/B ratio in disease susceptibility.

Association among nocturnal enuresis, body weight and obstructive sleep apnea in children of south Italy: an observational study.

BACKGROUND: To evaluate the rate of nocturnal enuresis (NE), body weight and obstructive sleep apnea in children 5 to 10 years of age in South Italy and the possible association among these disorders. METHODS: We have administered 1100 validated questionnaires, in Italian language, to parents and we have analyzed data with a logistic regression. RESULTS: Forty-two percent of children had a BMI≥85th (group 1) vs 58.0% normal weight children at the same age (group 2). There is a higher number of overweight males compared to females without statistically differences. In group 1 there was a higher number of children with NE and obstructive sleep disorders and some children present with the association among these three disorders. CONCLUSIONS: There are no statistically differences between two study groups for the association body weight-NE, body weight-NE-obstructive sleep disorders.

Storage and stability of IgG and IgM monoclonal antibodies dried on filter paper and utility in Neisseria meningitidis serotyping by Dot-blot ELISA.

BACKGROUND: A simple filter paper method was developed for, the transport and storage of monoclonal antibodies (Mabs) at room temperature or -20 degrees C after spotting on filter paper, for subsequent serotyping of outer membrane antigens of N.meningitidis by dot-blot ELISA. METHODS: Monoclonal antibodies (Mabs) were spotted within a 0.5-1 cm diameter area of Whatman grade 903 paper, which were stored individually at room temperature or at -20 degrees C. These MAbs were stored and analyzed after periods of one week, 4 weeks, 12 months, or 13 years in the case of frozen Mab aliquots, or after 4 weeks at -20 degrees C or at room temperature (RT) in the case of Mabs dried on filter paper strips. Assays were performed in parallel using dot-blot ELISA. In addition to the MAbs specific for serotyping class 1, 2 or 3, we used a larger number of Mabs for polysaccharides, lipooligosaccharides (LOS), class 5 and cross-reactive antigens for native outer membrane of N.meningitidis. The Mabs dried on filter paper were eluted with phosphate-buffered saline (PBS) containing 0.2% gelatin. RESULTS: Mabs of the isotypes IgG and IgM dried on filter papers were not affected by duration of storage. The detection by serotyping Mabs was generally consistent for dried filter paper MAb samples stored frozen for over 1 year at -20 degrees C, and although decreased reactive antibody titers were found after storage, this did not interfere with the specificity of the Mabs used after 13 years as dry spots on filter paper. CONCLUSION: The use of filter paper is an inexpensive and convenient method for collecting, storing, and transporting Mab samples for serotyping studies. In addition, the samples occupy little space and can be readily transported without freezing. The efficiency of using immunoglobulin G (IgG) or M (IgM) eluted was found to be consistent with measurement of IgG or IgM titers in most corresponding, ascites Mabs stored frozen for over 1 year. The application of meningococcal typing methods and designations depend on the question being asked.

Production of monoclonal antibodies against Neisseria meningitidis using popliteal lymph nodes and in vivo/in vitro immunization: prevalence study of new monoclonal antibodies in greater São Paulo, Brazil.

A rapid and efficient method for preparing monoclonal antibody (MAb) serotypes using Neisseria meningitidis outer membrane were used in BALB/c mouse footpads for the immunization. The popliteal lymph nodes were isolated 19 days later for MAb-producing hybridomas, from which the MAbs against the 37 kDa protein were screened. Variations in class 2/3 (PorB) proteins form the basis for meningococcal serotyping. This is the first report on the preparation of MAbs against N. meningitidis that is specific to PorB protein using popliteal lymph nodes. The new monoclonal antibodies were specific for PorB outer membrane protein FL24(PL)Br, a new serotype 24 class 3 antigens of non-typeable (NT:NST) serogroup B strain, and FL14(PL)Br specific for the serotype 14, and reacted with the S3446 reference strain analyzed. A total of 12% of the case isolates reacted with one or more of the monoclonal antibodies. The high-affinity MAbs produced by hybridoma methodology provide a basis for further research on the pathogenesis and early diagnosis of meningococcus.