Case Report: Morphologic and Functional Characteristics of Intestinal Mucosa in a Child With Short Bowel Syndrome After Treatment With Teduglutide: Evidence in Favor of GLP-2 Analog Safety

Teduglutide is a glucagon-like peptide-2 (GLP-2) analog employed in patients with short bowel syndrome (SBS) to reduce the need of parenteral nutrition in these patients, by virtue of its effects on enteric function. The experimental studies reported that the stimulating action of GLP-2 on epithelial turnover implies the potential development of dysplastic and neoplastic lesion. However, the clinical trials could not detect preneoplastic lesions on histologic material, and in a recent pilot study the occurrence of polyps was similar before and after treatment and included only low-grade dysplastic lesions. Another clue in GLP-2 function in stimulating mucosal restore is its enhancement through cooperation with epidermal growth factor (EGF). In this study, we analyzed gastroscopy and colonoscopy samplings from a child successfully weaned off parenteral nutrition with teduglutide. Villous and crypt structure was regular both in duodenal and in colonic samplings; in properly oriented villi, villus/crypt ratio was regular. The absorptive epithelium demonstrated a regular morphology. No atypia was detected in enterocytes, along epithelial structures. At the ultrastructural analysis, only a few enterocytes with vacuolized cytoplasm were observed. An S-phase marker Ki67 stained nuclei in the transitional amplifying zone, while nuclei stained by the cell cycle regulatory proteins p21 and p27 were placed in the differentiated epithelium of the duodenal villi and colonic crypts, as in the control cases. The counts of enterocytes immunostained with the same antisera, evaluated with image analysis software, were in the range of control cases. The ratio of the number of epidermal growth factor receptor (EGFR) signals/the number of centromere probe of chromosome 7 (CEP7) signals was less than 2. The findings available from this single patient are consistent with good preservation of functional capability of intestinal epithelium after treatment with GLP-2, given the histologic and ultrastructural features of enterocytes. In addition, the findings from cell cycle regulatory proteins immunolocalization and quantitative analysis show that cell renewal machinery in our case is comparable to control cases. The gene of the receptor EGFR is regularly expressed in enteric epithelium of our case. Morphologic and functional data from our patient improve evidence in favor of the safety of GLP-2 employ in SBS

Apoptotic epitope-specific CD8+ T cells and interferon signaling intersect in chronic hepatitis C virus infection

CD8(+) T cells specific to caspase-cleaved antigens derived from apoptotic T cells represent a principal player in chronic immune activation (CIA). Here, we found that both apoptotic epitope (AE)-specific and hepatitis C virus (HCV)-specific CD8(+) T cells were mostly confined within the effector memory (EM) or terminally differentiated EM CD45RA(+) cell subsets expressing a dysfunctional T-helper-1-like signature program in chronic (c)HCV infection. However, AE-specific CD8(+) T cells produced tumor necrosis factor (TNF)-α and interleukin-2 at the intrahepatic level significantly more than HCV-specific CD8(+) T cells, despite both populations acquiring high levels of programmed death-1 receptor expression. Contextually, only AE-specific CD8(+) T cells correlated with both interferon-stimulated gene levels in T cells and hepatic fibrosis score. Taken together, these data suggest that AE-specific CD8(+) T cells can sustain CIA by their capacity to produce TNF-α and be resistant to inhibitory signals more than HCV-specific CD8(+) T cells in cHCV infection

Implications of Storing Urinary DNA from Different Populations for Molecular Analyses

Molecular diagnosis using urine is established for many sexually transmitted diseases and is increasingly used to diagnose tumours and other infectious diseases. Storage of urine prior to analysis, whether due to home collection or bio-banking, is increasingly advocated yet no best practice has emerged. Here, we examined the stability of DNA in stored urine in two populations over 28 days.Urine from 40 (20 male) healthy volunteers from two populations, Italy and Zambia, was stored at four different temperatures (RT, 4 degrees C, -20 degrees C & -80 degrees C) with and without EDTA preservative solution. Urines were extracted at days 0, 1, 3, 7 and 28 after storage. Human DNA content was measured using multi-copy (ALU J) and single copy (TLR2) targets by quantitative real-time PCR. Zambian and Italian samples contained comparable DNA quantity at time zero. Generally, two trends were observed during storage; no degradation, or rapid degradation from days 0 to 7 followed by little further degradation to 28 days. The biphasic degradation was always observed in Zambia regardless of storage conditions, but only twice in Italy.Site-specific differences in urine composition significantly affect the stability of DNA during storage. Assessing the quality of stored urine for molecular analysis, by using the type of strategy described here, is paramount before these samples are used for molecular prognostic monitoring, genetic analyses and disease diagnosis

IP-10 detection in urine is associated with lung diseases

<p>Abstract</p> <p>Background</p> <p>blood cytokines and chemokines have been proposed as biomarkers for tuberculosis (TB). Recently, some immune mediators found in the urine of patients with renal dysfunctions have also been suggested as potential biomarkers. Finding biomarkers for TB in urine would present several advantages over blood in terms of collection and safety. The objective of this study was to investigate the presence of cytokines and chemokines in the urine of patients with pulmonary TB at the time of diagnosis. In a subgroup, the evaluation was also performed during TB treatment and at therapy completion. Patients with lung diseases other than TB, and healthy subjects were also enrolled.</p> <p>Methods</p> <p>urine samples from 138 individuals, after exclusion of renal dysfunctions, were collected during an 18 month-period. Among them, 58 received a diagnosis of pulmonary TB, 28 resulted having lung diseases other than TB, and 34 were healthy subjects. Moreover, 18 TB patients, 9 of whom were tested 2 months after AFB smear sputum reversion and 9 of whom were cured of TB were also included. Cytokines and chemokines in urine were evaluated using a Cytometric-Bead-Array-Flex-Set. IP-10 detection in 49 subjects was also carried out in parallel by using an Enzyme Linked ImmunoSorbent Assay (ELISA).</p> <p>Results</p> <p>IFN-γ, TNF-α, IL-2, IL-8, MIP-1α, MIP-1β and RANTES were poorly detected in all urine samples. Conversely, IP-10 was consistently detected in urine and its level was significantly increased in patients with lung disease compared to healthy subjects (p < 0.001). Increased IP-10 levels were found in both pulmonary TB and lung diseases other than TB. Moreover lower IP-10 levels were found in cured-TB patients compared to the levels at the time of diagnosis, and this difference was close to significance (p = 0.06). Interestingly, we demonstrated a significant correlation between the data obtained by flow cytometry and ELISA (r²0.82, p < 0.0001).</p> <p>Conclusions</p> <p>IP-10, in contrast to IFN-γ, TNF-α, IL-2, IL-8, MIP-1α, MIP-1β and RANTES, is detectable in the urine of patients with pulmonary diseases in the absence of renal dysfunctions. Moreover, the IP-10 level in cured-TB patients is comparable to that found in healthy subjects. More studies are needed to further investigate the clinical utility of these findings.</p

Burnout among surgeons before and during the SARS-CoV-2 pandemic: an international survey

Background: SARS-CoV-2 pandemic has had many significant impacts within the surgical realm, and surgeons have been obligated to reconsider almost every aspect of daily clinical practice. Methods: This is a cross-sectional study reported in compliance with the CHERRIES guidelines and conducted through an online platform from June 14th to July 15th, 2020. The primary outcome was the burden of burnout during the pandemic indicated by the validated Shirom-Melamed Burnout Measure. Results: Nine hundred fifty-four surgeons completed the survey. The median length of practice was 10&nbsp;years; 78.2% included were male with a median age of 37&nbsp;years old, 39.5% were consultants, 68.9% were general surgeons, and 55.7% were affiliated with an academic institution. Overall, there was a significant increase in the mean burnout score during the pandemic; longer years of practice and older age were significantly associated with less burnout. There were significant reductions in the median number of outpatient visits, operated cases, on-call hours, emergency visits, and research work, so, 48.2% of respondents felt that the training resources were insufficient. The majority (81.3%) of respondents reported that their hospitals were included in the management of COVID-19, 66.5% felt their roles had been minimized; 41% were asked to assist in non-surgical medical practices, and 37.6% of respondents were included in COVID-19 management. Conclusions: There was a significant burnout among trainees. Almost all aspects of clinical and research activities were affected with a significant reduction in the volume of research, outpatient clinic visits, surgical procedures, on-call hours, and emergency cases hindering the training. Trial registration: The study was registered on clicaltrials.gov "NCT04433286" on 16/06/2020

Impact of bacterial motility on biosorption and cometabolism of pyrene in a porous medium

8 páginas.- 6 figuras.- 40 referenciasThe risks of pollution by polycyclic aromatic hydrocarbons (PAHs) may increase in bioremediated soils as a result of the formation of toxic byproducts and the mobilization of pollutants associated to suspended colloids. In this study, we used the motile and chemotactic bacterium Pseudomonas putida G7 as an experimental model for examining the potential role of bacterial motility in the cometabolism and biosorption of pyrene in a porous medium. For this purpose, we conducted batch and column transport experiments with 14C-labelled pyrene loaded on silicone O-rings, which acted as a passive dosing system. In the batch experiments, we observed concentrations of the 14C-pyrene equivalents well above the equilibrium concentration observed in abiotic controls. This mobilization was attributed to biosorption and cometabolism processes occurring in parallel. HPLC quantification revealed pyrene concentrations well below the 14C-based quantifications by liquid scintillation, indicating pyrene transformation into water-soluble polar metabolites. The results from transport experiments in sand columns revealed that cometabolic-active, motile cells were capable of accessing a distant source of sorbed pyrene. Using the same experimental system, we also determined that salicylate-mobilized cells, inhibited for pyrene cometabolism, but mobilized due to their tactic behavior, were able to sorb the compound and mobilize it by biosorption. Our results indicate that motile bacteria active in bioremediation may contribute, through cometabolism and biosorption, to the risk associated to pollutant mobilization in soils. This research could be the starting point for the development of more efficient, low-risk bioremediation strategies of poorly bioavailable contaminants in soils. © 2020 Elsevier B.V.We thank the Spanish Ministry of Economy, Industry and Competitiveness (CGL2016-77497-R), the Andalusian Government (RNM 2337), and the European Commission (LIFE15 ENV/IT/000396). Joaquim Vila is a Serra Húnter Fellow (Generalitat de Catalunya).Peer reviewe

Characterization of black patina from the Tiber River embankments using Next-Generation Sequencing.

Black patinas are very common biological deterioration phenomena on lapideous artworks in outdoor environments. These substrates, exposed to sunlight, and atmospheric and environmental agents (i.e. wind and temperature changes), represent extreme environments that can only be colonized by highly versatile and adaptable microorganisms. Black patinas comprise a wide variety of microorganisms, but the morphological plasticity of most of these microorganisms hinders their identification by optical microscopy. This study used Next-Generation Sequencing (NGS) (including shotgun and amplicon sequencing) to characterize the black patina of the travertine embankments (muraglioni) of the Tiber River in Rome (Italy). Overall, the sequencing highlighted the rich diversity of bacterial and fungal communities and allowed the identification of more than one hundred taxa. NGS confirmed the relevance of coccoid and filamentous cyanobacteria observed by optical microscopy and revealed an informative landscape of the fungal community underlining the presence of microcolonial fungi and phylloplane yeasts. For the first time high-throughput sequencing allowed the exploration of the expansive diversity of bacteria in black patina, which has so far been overlooked in routine analyses. Furthermore, the identification of euendolithic microorganisms and weathering agents underlines the biodegradative role of black patina, which has often been underestimated. Therefore, the use of NGS to characterize black patinas could be useful in choosing appropriate conservation treatments and in the monitoring of stone colonization after the restoration interventions

Proteomic analysis reveals a major role for contact inhibition in the terminal differentiation of hepatocytes

Hepatocytes are considered an exception of the paradigmatic inverse correlation between cell proliferation and terminal differentiation. In fact, hepatic vital functions are guaranteed by proliferating parenchymal cells during liver regeneration. However, a fine molecular characterization of the relationship between proliferation and differentiation in hepatocytes has been hampered by the lack of reliable in vivo or in vitro models

ANTI CAPSID DRUGS HAP12 AND AT130 TARGET HBV CORE PROTEIN NUCLEAR FUNCTIONS

Introduction and aim: HBV core protein (Cp) represents an attractive new therapeutic target for HBV chronic infection. Cp has been shown to bind the nuclear cccDNA mini-chromosome as well as a number of cellular genes promoters. Several compounds that target Cp and HBV capsids assembly, including the hetero- aryl-dihydropyrimidines (HAPs) and the phenyl-propenamide derivatives AT61 and AT130, have been shown to inhibit HBV repli- cation in vitro and in vivo. HAPs and AT130 enhance the rate of Cp assembly and stabilize preferentially non-capsid polymers of Cp. Here we investigated the ability of the Core protein Assembly Mod- ulators (CaMPs) HAP12 and AT130 to affect both nuclear cccDNA transcription and cytoplasmic capsid assembly Cp functions. Methods: HAP12 and AT130 effects on capsid-associated HBV- DNA, cccDNA and pgRNA levels (quantitative real-time PCR with specific primers) were assessed in: (a) HBV-infected NTCP-HepG2 cells; (b) AD38 inducible HBV stable cell line. Recruitment of HBc and histone modifications on the viral minichromosome were assessed using the cccDNA ChIP assay in AD38 cells. Results: CaMPs treatments resulted in a very strong inhibition of HBV replication (>95%) and a significant but incomplete reduction of the stable cccDNA pool. A strong effect on cccDNA-dependent HBeAg production (AD38 tet-off) and pgRNA transcription (AD38 tet-off/tet-on and NTCP-HepG2 infected cells) was also demon- strated. The ability of HAP12 to target cccDNA transcription was confirmed by the reduced cccDNA-bound H3 histone acetylation and the decreased HBc occupancy on the cccDNA in induced AD38 cells. Importantly, when CaMPs treatment was started during infec- tion, cccDNA formation/accumulation was completely inhibited (>95%) and viral replication was blunted. Conclusions: Anti-capsid compounds (CpAMs) have an impact on Cp nuclear functions at multiple levels: block of new cccDNA formation/accumulation, reduction of an established cccDNA pool and inhibition of HBc occupancy and histone acetylation on the cccDNA that translate into a reduced pgRNA transcription