Counteracting gemcitabine+nab-paclitaxel induced dysbiosis in KRAS wild type and KRASG12D mutated pancreatic cancer in vivo model

Pancreatic cancer (PC) has a very low survival rate mainly due to late diagnosis and refractoriness to therapies. The latter also cause adverse effects negatively affecting the patients' quality of life, often requiring dose reduction or discontinuation of scheduled treatments, compromising the chances of cure. We explored the effects of a specific probiotic blend on PC mice xenografted with KRAS wild-type or KRASG12D mutated cell lines alone or together with gemcitabine+nab-paclitaxel treatment to then assess tumor volume and clinical pathological variables. Beside a semi-quantitative histopathological evaluation of murine tumor and large intestine samples, histochemical and immunohistochemical analyses were carried out to evaluate collagen deposition, proliferation index Ki67, immunological microenvironment tumor-associated, DNA damage markers and also mucin production. Blood cellular and biochemical parameters and serum metabolomics were further analyzed. 16S sequencing was performed to analyze the composition of fecal microbiota. Gemcitabine+nab-paclitaxel treatment impaired gut microbial profile in KRAS wild-type and KRASG12D mice. Counteracting gemcitabine+nab-paclitaxel- induced dysbiosis through the administration of probiotics ameliorated chemotherapy side effects and decreased cancer-associated stromatogenesis. Milder intestinal damage and improved blood count were also observed upon probiotics treatment as well as a positive effect on fecal microbiota, yielding an increase in species richness and in short chain fatty acids producing- bacteria. Mice' serum metabolomic profiles revealed significant drops in many amino acids upon probiotics administration in KRAS wild-type mice while in animals transplanted with PANC-1 KRASG12D mutated all treated groups showed a sharp decline in serum levels of bile acids with respect to control mice. These results suggest that counteracting gemcitabine+nab-paclitaxel-induced dysbiosis ameliorates chemotherapy side effects by restoring a favorable microbiota composition. Relieving adverse effects of the chemotherapy through microbiota manipulation could be a desirable strategy in order to improve pancreatic cancer patients' quality of life and to increase the chance of cure

Goodbye Hartmann trial: a prospective, international, multicenter, observational study on the current use of a surgical procedure developed a century ago

Background: Literature suggests colonic resection and primary anastomosis (RPA) instead of Hartmann's procedure (HP) for the treatment of left-sided colonic emergencies. We aim to evaluate the surgical options globally used to treat patients with acute left-sided colonic emergencies and the factors that leading to the choice of treatment, comparing HP and RPA. Methods: This is a prospective, international, multicenter, observational study registered on ClinicalTrials.gov. A total 1215 patients with left-sided colonic emergencies who required surgery were included from 204 centers during the period of March 1, 2020, to May 31, 2020. with a 1-year follow-up. Results: 564 patients (43.1%) were females. The mean age was 65.9 ± 15.6 years. HP was performed in 697 (57.3%) patients and RPA in 384 (31.6%) cases. Complicated acute diverticulitis was the most common cause of left-sided colonic emergencies (40.2%), followed by colorectal malignancy (36.6%). Severe complications (Clavien-Dindo ≥ 3b) were higher in the HP group (P < 0.001). 30-day mortality was higher in HP patients (13.7%), especially in case of bowel perforation and diffused peritonitis. 1-year follow-up showed no differences on ostomy reversal rate between HP and RPA. (P = 0.127). A backward likelihood logistic regression model showed that RPA was preferred in younger patients, having low ASA score (≤ 3), in case of large bowel obstruction, absence of colonic ischemia, longer time from admission to surgery, operating early at the day working hours, by a surgeon who performed more than 50 colorectal resections. Conclusions: After 100 years since the first Hartmann's procedure, HP remains the most common treatment for left-sided colorectal emergencies. Treatment's choice depends on patient characteristics, the time of surgery and the experience of the surgeon. RPA should be considered as the gold standard for surgery, with HP being an exception

Production of He-4 and (4) in Pb-Pb collisions at root(NN)-N-S=2.76 TeV at the LHC

Results on the production of He-4 and (4) nuclei in Pb-Pb collisions at root(NN)-N-S = 2.76 TeV in the rapidity range vertical bar y vertical bar <1, using the ALICE detector, are presented in this paper. The rapidity densities corresponding to 0-10% central events are found to be dN/dy4(He) = (0.8 +/- 0.4 (stat) +/- 0.3 (syst)) x 10(-6) and dN/dy4 = (1.1 +/- 0.4 (stat) +/- 0.2 (syst)) x 10(-6), respectively. This is in agreement with the statistical thermal model expectation assuming the same chemical freeze-out temperature (T-chem = 156 MeV) as for light hadrons. The measured ratio of (4)/He-4 is 1.4 +/- 0.8 (stat) +/- 0.5 (syst). (C) 2018 Published by Elsevier B.V.Peer reviewe

Accurate and precise DNA quantification in the presence of different amplification efficiencies using an improved Cy0 method.

Quantitative real-time PCR represents a highly sensitive and powerful technology for the quantification of DNA. Although real-time PCR is well accepted as the gold standard in nucleic acid quantification, there is a largely unexplored area of experimental conditions that limit the application of the Ct method. As an alternative, our research team has recently proposed the Cy0 method, which can compensate for small amplification variations among the samples being compared. However, when there is a marked decrease in amplification efficiency, the Cy0 is impaired, hence determining reaction efficiency is essential to achieve a reliable quantification. The proposed improvement in Cy0 is based on the use of the kinetic parameters calculated in the curve inflection point to compensate for efficiency variations. Three experimental models were used: inhibition of primer extension, non-optimal primer annealing and a very small biological sample. In all these models, the improved Cy0 method increased quantification accuracy up to about 500% without affecting precision. Furthermore, the stability of this procedure was enhanced integrating it with the SOD method. In short, the improved Cy0 method represents a simple yet powerful approach for reliable DNA quantification even in the presence of marked efficiency variations

Scienza Politica - Guide agli studi di scienze sociali in Italia

Tra filosofia politica e teoria giuridica, la scienza politica ha dovuto interrogarsi sull'autonomia del proprio oggetto e metodo, come testimonia il difficile affrancamento di essa dall'alveo delle scienze sociali. In tale prospettiva, gli studi raccolti nel volume forniscono un apporto all'interpretazione storica della disciplina.- Indice #6- Presentazione, Marcello Pacini #10- Premessa #16- Introduzione. Ancora un bilancio lamentevole?, Leonardo Morlino #18- Cap.I Teoria e macropolitica, Leonardo Morlino #66- Cap.II La cultura politica, Giacomo Sani #102- Cap.III Le strutture di rappresentanza, Angelo Panebianco #120- Cap.IV Elezioni e comportamento elettorale, Renato Mannheimer #158- Cap.V Le élites politiche, Mauro Calise #194- Cap.VI Strutture e processi decisionali, Carlo Guarnieri #212- Cap.VII Burocrazia e magistratura, Carlo Guarnieri #236- Cap.VII Le politiche pubbliche, Maurizio Ferrera #254- Appendice Bibliografica, Franco Mattei #27

Effect of increasing PCR inhibition on the accuracy of the HE and LE amplification systems.

<p>PCR quantifications were performed using HE and LE systems in the presence of an equal starting number of template molecules and increasing blocked primer concentrations. Panels (A) and (B) show the obtained amplification plots for each system. Panels (C) and (D) show the relative errors obtained using <i>Cy0</i> and <i>Cy0</i> corrected following Eq. 10. The relative error was reported as <i>Log(DNA_exp)</i>–<i>Log(DNA_obs)</i> where <i>Log(DNA_exp)</i> is the number of expected molecules and <i>Log(DNA_obs)</i> is the number of calculated molecules using the <i>Cy0 (Black dots)</i> or <i>Cy0_corrected (White dots).</i> Each symbol represents a mean 6 runs. SOD detected almost all the runs in which the blocked primer was added as outliers.</p

Description of the High Efficiency (HE) and Low Efficiency (LE) systems.

<p>The HE and LE systems were obtained amplifying the mitochondrial MT-ND1 gene with the same reverse primer ND1R2 but using ND1F2 or ND1F5 as forward primer, respectively. Panel (A) shows the primer annealing locations in the mitochondrial sequence; panel (B) shows the secondary structure of the obtained amplification products calculated using <a href="http://mfold.rna.albany.edu/" target="_blank">http://mfold.rna.albany.edu/</a>. This analysis shows that ND1F5 primer overlaps the stem-loop at 5′ terminus, The overlapping coupled with its reduced length impairs the primer’s ability to bind the DNA template. Panel (C) shows the overall efficiency analysis of the two systems (HE and LE). The slopes of the two regressions are significantly different, with slope_HE = −3.488; slope_LE = −3.942; consequently, the overall efficiencies, calculated using the equation , were <i>E_overall-HE</i> = 0.93; <i>E_overall-LE</i> = 0.79.</p

Real-time PCR quantifications in the presence of “Competimer.”

<p>The <i>Cy0</i> and <i>Cy0_corrected</i> values were calculated from the independent freely available data set named “Competimer”, reported in Ruijter et al. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0068481#pone.0068481-Ruijter1" target="_blank">[32]</a>. Briefly, the amplification curves were obtained using decreasing input gDNA amounts (64, 16, 4, 1, 0.25 and 0.0625 ng/µl) and in optimal (% Competimer = 0–5) and inhibited (increasing % Competimer from 10 to 50%) amplification conditions. The and parameters were estimated from optimal amplification conditions. Panel (A) shows <i>Cy0</i> and <i>Cy0_corrected</i> values obtained for each condition; Panel (B) shows the standard deviation calculated from the Cqs (<i>Cy0</i> and <i>Cy0_corrected</i>) corresponding to the same input cDNA quantity.</p