Analysis of lactase processing in rabbit

AbstractThe proteolytic processing of rabbit intestinal lactase-phlorizin-hydrolase (LPH) was studied by pulse-chase and continuous labeling experiments in organ culture from 15-day-old rabbits in the presence of glycosylation and processing inhibitors. Monensin and brefeldin A inhibited the two proteolytic cleavages of the precursor indicating that they are post-Golgi events as previously reported for the unique cleavage of LPH in man [1]. The inhibition was not related to a concomitant alteration glycosylation; in fact, if trimming was blocked by MDNM the abnormal glycosylated precursor was proteolytically processed normally. Finally the use of the anti-microtubular drug colchicine strongly inhibited both cleavages and caused accumulation of the complex-glycosylated precursor form in the brush border fraction indicating that proteolytic events depend on intact microtubule (transport)

Potential Celiac Patients: A Model of Celiac Disease Pathogenesis

BACKGROUND AND AIM: Potential celiacs have the 'celiac type' HLA, positive anti-transglutaminase antibodies but no damage at small intestinal mucosa. Only a minority of them develops mucosal lesion. More than 40 genes were associated to Celiac Disease (CD) but we still do not know how those pathways transform a genetically predisposed individual into an affected person. The aim of the study is to explore the genetic features of Potential CD individuals. METHODS: 127 'potential' CD patients entered the study because of positive anti-tissue transglutaminase and no mucosal lesions; about 30% of those followed for four years become frankly celiac. They were genotyped for 13 polymorphisms of 'candidate genes' and compared to controls and celiacs. Moreover, 60 biopsy specimens were used for expression studies. RESULTS: Potential CD bear a lighter HLA-related risk, compared to celiac (??(2)???=???48.42; p value???=???1×10(-8)). They share most of the polymorphisms of the celiacs, but the frequency of c-REL* G allele was suggestive for a difference compared to celiac (??(2)???=???5.42; p value???=???0.02). One marker of the KIAA1109/IL-2/IL-21 candidate region differentiated potentials from celiac (rs4374642: ??2???=???7.17, p value???=???0.01). The expression of IL-21 was completely suppressed in potentials compared to celiacs (p value???=???0.02) and to controls (p value???=???0.02), in contrast IL-2, KIAA1109 and c-REL expression were over-expressed. CONCLUSIONS: Potential CD show genetic features slightly different from celiacs. Genetic and expression markers help to differentiate this condition. Potential CD is a precious biological model of the pathways leading to the small intestinal mucosal damage in genetically predisposed individuals

Tungiasis: Case Report of a Traveller to Kenya

Tungiasis is a neglected parasitic skin disease caused by the permanent penetration of the female sand flea Tunga penetrans (also called jigger flea) into the skin of its host. Growing urbanisation, improved housing and the use of appropriate footwear have presumably led to an overall reduction of the occurrence of this ectoparasitosis within the last few decades. However, it is still highly prevalent in regions where people live in extreme poverty, such as in many Latin American and African countries [1, 2]. We report the case of a 44-year-old woman who returned from an excursion trip to Kenya's savannah with an infection of T. penetrans located on her right big toe around the nail. The natural history, pathology, epidemiology, diagnosis, therapy and control of this parasitic skin disease are discussed [1]

Deep Generative Models: The winning key for large and easily accessible ECG datasets?

Large high-quality datasets are essential for building powerful artificial intelligence (AI) algorithms capable of supporting advancement in cardiac clinical research. However, researchers working with electrocardiogram (ECG) signals struggle to get access and/or to build one. The aim of the present work is to shed light on a potential solution to address the lack of large and easily accessible ECG datasets. Firstly, the main causes of such a lack are identified and examined. Afterward, the potentials and limitations of cardiac data generation via deep generative models (DGMs) are deeply analyzed. These very promising algorithms have been found capable not only of generating large quantities of ECG signals but also of supporting data anonymization processes, to simplify data sharing while respecting patients' privacy. Their application could help research progress and cooperation in the name of open science. However several aspects, such as a standardized synthetic data quality evaluation and algorithm stability, need to be further explored

Short wheat challenge is a reproducible in-vivo assay to detect immune response to gluten.

It has been reported that interferon (IFN)-γ-secreting T cells reactive to gluten can be detected in the peripheral blood of individuals with treated coeliac disease (CD) after a short consumption of wheat-containing food. By contrast, very little is known about the reproducibility of this in-vivo procedure in the same patient cohort which underwent two, or more, gluten consumptions. Fourteen coeliac patients in remission consumed wheat bread for 3 days; 13 underwent a second gluten challenge after a wash-out of 3-10 months on a strict gluten-free diet. Immune reactivity to gluten was analysed in peripheral blood by detecting IFN-γ before and 6 days after commencing a gluten diet. Gliadin-specific IFN-γ-secreting CD4(+) T cells increased significantly on day 6 of the first challenge. These cells resulted as prevalently human leucocyte antigen (HLA)-DQ restricted and with a phenotype of gut homing, as suggested by the expression of β7-integrin. Similarly, reactiveness to gliadin was observed after the second wheat consumption, although with an individual variability of responses at each challenge. Our findings confirmed that the short wheat challenge is a non-invasive approach to investigate the gluten-related immune response in peripheral blood of subjects intolerant to gluten. Furthermore, we demonstrated that the in-vivo procedure can be reproduced in the same subject cohort after a gluten wash-out of at least 3 months. Our study has important implications for the application of this procedure to clinical practice

In vitro-deranged intestinal immune response to gliadin in type 1 diabetes.

Dietary gluten has been associated with an increased risk of type 1 diabetes. We have evaluated inflammation and the mucosal immune response to gliadin in the jejunum of patients with type 1 diabetes. Small intestinal biopsies from 17 children with type 1 diabetes without serological markers of celiac disease and from 50 age-matched control subjects were examined by immunohistochemistry. In addition, biopsies from 12 type 1 diabetic patients and 8 control subjects were cultured with gliadin or ovalbumin peptic-tryptic digest and examined for epithelial infiltration and lamina propria T-cell activation. The density of intraepithelial CD3(+) and gammadelta(+) cells and of lamina propria CD25(+) mononuclear cells was higher in jejunal biopsies from type 1 diabetic patients versus control subjects. In the patients' biopsies cultured with peptic-tryptic gliadin, there was epithelial infiltration by CD3(+) cells, a significant increase in lamina propria CD25(+) and CD80(+) cells and enhanced expression of lamina propria CD54 and crypt HLA-DR. No such phenomena were observed in control subjects, even those with celiac disease-associated HLA haplotypes. In conclusion, signs of mucosal inflammation were present in jejunal biopsies from type 1 diabetic patients, and organ culture studies indicate a deranged mucosal immune response to gliadin

Combined Analysis of Methylation and Gene Expression Profiles in Separate Compartments of Small Bowel Mucosa Identified Celiac Disease Patients' Signatures

By GWAS studies on celiac disease, gene expression was studied at the level of the whole intestinal mucosa, composed by two different compartments: epithelium and lamina propria. Our aim is to analyse the gene-expression and DNA methylation of candidate genes in each of these compartments. Epithelium was separated from lamina propria in biopsies of CeD patients and CTRs using magnetic beads. Gene-expression was analysed by RT-PC; methylation analysis required bisulfite conversion and NGS. Reverse modulation of gene-expression and methylation in the same cellular compartment was observed for the IL21 and SH2B3 genes in CeD patients relative to CTRs. Bioinformatics analysis highlighted the regulatory elements in the genomic region of SH2B3 that altered methylation levels. The cREL and TNFAIP3 genes showed methylation patterns that were significantly different between CeD patients and CTRs. In CeD, the genes linked to inflammatory processes are up-regulated, whereas the genes involved in the cell adhesion/ integrity of the intestinal barrier are down-regulated. These findings suggest a correlation between gene-expression and methylation profile for the IL21 and SH2B3 genes. We identified a "gene-expression phenotype" of CeD and showed that the abnormal response to dietary antigens in CeD might be related not to abnormalities of gene structure but to the regulation of molecular pathways.This work was funded by the FC-Grant2013. Programma di mobilita nell'ambito delle reti di eccellenza POR Campania FSE 2007-2013 Asse V. The authors thank the European Laboratory for Food-induced Disease (ELFID). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Consistency in Polyclonal T-cell Responses to Gluten between Children and Adults with Celiac Disease

BACKGROUND & AIMS: Developing antigen-specific approaches for diagnosis and treatment of celiac disease requires a detailed understanding of the specificity of T cells for gluten. The existing paradigm is that T-cell lines and clones from children differ from those of adults in the hierarchy and diversity of peptide recognition. We aimed to characterize the T-cell response to gluten in children vs adults with celiac disease. METHODS: Forty-one children with biopsy-proven celiac disease (median age, 9 years old; 17 male), who had been on strict gluten-free diets for at least 3 months, were given a 3-day challenge with wheat; blood samples were collected and gluten-specific T cells were measured. We analyzed responses of T cells from these children and from 4 adults with celiac disease to a peptide library and measured T-cell receptor bias. We isolated T-cell clones that recognized dominant peptides and assessed whether gluten peptide recognition was similar between T-cell clones from children and adults. RESULTS: We detected gluten-specific responses by T cells from 30 of the children with celiac disease (73%). T cells from the children recognized the same peptides that were immunogenic to adults with celiac disease; deamidation of peptides increased these responses. Age and time since diagnosis did not affect the magnitude of T-cell responses to dominant peptides. T-cell clones specific for dominant α- or ω-gliadin peptides from children with celiac disease had comparable levels of reactivity to wheat, rye, and barley peptides as T-cell clones from adults with celiac disease. The α-gliadin-specific T cells from children had biases in T-cell receptor usage similar to those in adults. CONCLUSIONS: T cells from children with celiac disease recognize similar gluten peptides as T cells from adults with celiac disease. The findings indicate that peptide-based diagnostics and therapeutics for adults may also be used for children. Copyright © 2015 AGA Institute. Published by Elsevier Inc. All rights reserved

An Utstein-based model score to predict survival to hospital admission: The UB-ROSC Score

Abstract Aims To develop and validate a multi-parametric practical score to predict the probability of survival to hospital admission of an out-of-hospital cardiac arrest (OHCA) victim by using Utstein Style-based variables. Methods All consecutive OHCA cases occurring from 2015 to 2017 in two regions, Pavia Province (Italy) and Canton Ticino (Switzerland) were included. We used random effect logistic regression to model survival to hospital admission after an OHCA. We computed the model area under the ROC curve (AUC ROC) for discrimination and we performed both internal and external validation by considering all OHCAs occurring in the aforementioned regions in 2018. The Utstein-Based ROSC (UB-ROSC) score was derived by using the coefficients estimated in the regression model. The score value was obtained adding the pertinent score components calculated for each variable. The score was then plotted against the probability of survival to hospital admission. Results 1962 OHCAs were included (62% male, mean age 73 ± 16 years). Age, aetiology, location, witnessed OHCA, bystander CPR, EMS arrival time and shockable rhythm were independently associated with survival to hospital admission. The model showed excellent discrimination (AUC 0.83, 95%CI 0.81–0.85) for predicting survival to hospital admission, also at internal cross-validation (AUC 0.82, 95%CI 0.80–0.84). The model maintained good discrimination after external validation by using the 2018 OHCA cohort (AUC 0.77, 95%CI 0.74–0.80). Conclusions UB-ROSC score is a novel score that predicts the probability of survival to hospital admission of an OHCA victim. UB-ROSC shall help in setting realistic expectations about sustained ROSC achievement during resuscitation manoeuvres

Teaching and learning human Anatomy in the University of Pavia: from models and clinical specimens to prosection on 3D models from our museum collection

Due to decline of resources and support for teaching human anatomy, in our Institute the use of cadaver dissection is not economically feasible. After a few years in which I was able to perform prosection on fixed organs belonging to the Institute collection, over the last years students learned topographical anatomy on commercial plastic models (bones, muscles, joints, trunks and brains). New perspectives recently occurred thanks to a collaboration with Prof. Auricchio’s group, which is involved in the strategic plan “3DPRINTING” (http://www. unipv.eu/site/home/area-stampa/articolo12952.html). First, we have segmented DICOM images of Computed Tomography (CT) to reconstruct 3D models of all the feet’s bones from a patient. Then, these 3D models have been post-processed to obtain suitable file for 3D printing. A 3DSYSTEMS ProJet 460 Plus, professional, full-color binder jetting printer (property of General Surgery2), has been used to create 3D models of feet’s bone by chalk powder binding. Medical students will use these models to test their own ability to recognize feet’s bones shape and to recompose them. Second, a plastic 3D anatomical model has been scanned by Artec Eva 3D Object Scanner to obtain a 3D virtual model of the physical one; this model has been modified to create a new modular model, printed with a process similar to one described above. Our Anatomy Institute is enriched by a Museum, established in the late eighteenth century by universally known anatomists (Rezia, Scarpa, Panizza, Zoja). This historical collection contains several sections (osteology, angiology, splanchnology, neurology and topographic anatomy). It is impossible to use these anatomical specimens of historical interest for prosection, but their life-size copies will constitute a cheap and effective method of learning. This strategy could not replace cadaver dissection experience but we hope that it could assist students in the comprehension of anatomical systems in a cost effective way within a systemic anatomy course. Besides, this method should optimize specimen’s choice and focalize student’s attention on peculiar, selected samples, preparing more appropriately medical students to their clinical practice