c-fos spontaneous expression during wakefulness is reversed during sleep in neuronal subsets of the rat cortex

c-fos is a transcriptional regulatory factor that can be rapidly and transiently induced in brain cells in response to a variety of stimuli. However, we observed that c-fos mRNA, in absence of exogenous stimulation, oscillates in different regions of the rat brain in relation to the rest-activity and sleep-wake circadian cycle (Grassi-Zucconi et al, 1993).This previous study was based on Northern blot analysis and did not provide any information on the neuronal cell populations subserving the circadian c-fos oscillation. Therefore, we used immuno- cytochemistry to detect in the rat brain the cells that express the Fos nuclear protein, encoded by the c-fos gene, during spontaneous wakefulness and sleep.The present report derives from analysis of the cerebral cortex; the findings observed at subcortical levels will be described elsewhere

Progression of Celiac Disease in Children With Antibodies Against Tissue Transglutaminase and Normal Duodenal Architecture

Potential celiac disease is characterized by positive results from serologic tests for tissue transglutaminase antibodies (anti-TG2) but normal duodenal architecture (Marsh stages 0-1). There is controversy over the best way to manage these patients. We investigated risk factors associated with the development of villous atrophy in children with potential celiac disease

Rotavirus VP6 protein mucosally delivered by cell wall-derived particles from Lactococcus lactis induces protection against infection in a murine model.

Rotaviruses are the primary cause of acute gastroenteritis in children worldwide. Although the implementation of live attenuated vaccines has reduced the number of rotavirus-associated deaths, variance in their effectiveness has been reported in different countries. This fact, among other concerns, leads to continuous efforts for the development of new generation of vaccines against rotavirus.In this work, we describe the obtention of cell wall-derived particles from a recombinant Lactococcus lactis expressing a cell wall-anchored version of the rotavirus VP6 protein. After confirming by SDS-PAGE, Western blot, flow cytometry and electronic immunomicroscopy that these particles were carrying the VP6 protein, their immunogenic potential was evaluated in adult BALB/c mice. For that, mucosal immunizations (oral or intranasal), with or without the dmLT [(double mutant Escherichia coli heat labile toxin LT(R192G/L211A)] adjuvant were performed. The results showed that these cell wall-derived particles were able to generate anti-rotavirus IgG and IgA antibodies only when administered intranasally, whether the adjuvant was present or not. However, the presence of dmLT was necessary to confer protection against rotavirus infection, which was evidenced by a 79.5 percent viral shedding reduction.In summary, this work describes the production of cell wall-derived particles which were able to induce a protective immune response after intranasal immunization. Further studies are needed to characterize the immune response elicited by these particles as well as to determine their potential as an alternative to the use of live L. lactis for mucosal antigen delivery

Clinical variability of neurofibromatosis 1: A modifying role of cooccurring PTPN11 variants and atypical brain MRI findings

Neurofibromatosis 1 (NF1) is a disorder characterized by variable expressivity caused by loss-of-function variants in NF1, encoding neurofibromin, a protein negatively controlling RAS signaling. We evaluated whether concurrent variation in proteins functionally linked to neurofibromin contribute to the variable expressivity of NF1. Parallel sequencing of a RASopathy gene panel in 138 individuals with molecularly confirmed clinical diagnosis of NF1 identified missense variants in PTPN11, encoding SHP2, a positive regulator of RAS signaling, in four subjects from three unrelated families. Three subjects were heterozygous for a gain-of-function variant and showed a severe expression of NF1 (developmental delay, multiple cerebral neoplasms and peculiar cortical MRI findings), and features resembling Noonan syndrome (a RASopathy caused by activating variants in PTPN11). Conversely, the fourth subject, who showed an attenuated presentation, carried a previously unreported PTPN11 variant that had a hypomorphic behavior in vitro. Our findings document that functionally relevant PTPN11 variants occur in a small but significant proportion of subjects with NF1 modulating disease presentation, suggesting a model in which the clinical expression of pathogenic NF1 variants is modified by concomitant dysregulation of protein(s) functionally linked to neurofibromin. We also suggest targeting of SHP2 function as an approach to treat evolutive complications of NF1

Clinical variability of neurofibromatosis 1: A modifying role of cooccurring PTPN11 variants and atypical brain MRI findings

Neurofibromatosis 1 (NF1) is a disorder characterized by variable expressivity caused by loss-of-function variants in NF1, encoding neurofibromin, a protein negatively controlling RAS signaling. We evaluated whether concurrent variation in proteins functionally linked to neurofibromin contribute to the variable expressivity of NF1. Parallel sequencing of a RASopathy gene panel in 138 individuals with molecularly confirmed clinical diagnosis of NF1 identified missense variants in PTPN11, encoding SHP2, a positive regulator of RAS signaling, in four subjects from three unrelated families. Three subjects were heterozygous for a gain-of-function variant and showed a severe expression of NF1 (developmental delay, multiple cerebral neoplasms and peculiar cortical MRI findings), and features resembling Noonan syndrome (a RASopathy caused by activating variants in PTPN11). Conversely, the fourth subject, who showed an attenuated presentation, carried a previously unreported PTPN11 variant that had a hypomorphic behavior in vitro. Our findings document that functionally relevant PTPN11 variants occur in a small but significant proportion of subjects with NF1 modulating disease presentation, suggesting a model in which the clinical expression of pathogenic NF1 variants is modified by concomitant dysregulation of protein(s) functionally linked to neurofibromin. We also suggest targeting of SHP2 function as an approach to treat evolutive complications of NF1

Procedure for Organic Matter Removal from Peat Samples for XRD Mineral Analysis

Ombrotrophic peatlands are recognized archives of past atmospheric mineral dust deposition. Net dust deposition rates, grain size, mineral hosts and source areas are typically inferred from down-core elemental data. Although elemental analysis can be time efficient and data rich, there are some inherent limitations. X-ray diffraction (XRD) analysis allowsdirect identification of mineral phases in environmental samples but few studies have applied this method to peat samples and a well-developed protocol for extracting the inorganic fraction of highly organic samples (>95%) is lacking. We tested and compared different levels of pre-treatment: no pre-treatment, thermal combustion (300, 350, 400, 450, 500 and 550 degrees C) and chemical oxidation (H2O2 and Na2S2O8) using a homogenised highly organic (>98%) composite peat sample. Subsequently, minerals were identified by XRD. The results show that combustion is preferred to chemical oxidation because it most efficiently removes organic matter (OM), an important pre-requisite for identifying mineral phases by XRD analysis. Thermally induced phase transitions can be anticipated when temperature is the only factor to take into consideration. Based on the data required in this studythe recommended combustion temperature is 500 degrees C which efficiently removes OM while preserving a majority of common dust minerals