Characterization of genetic biodiversity with Vitis vinifera L. Sangiovese and Colorino genotypes by AFLP and ISTR DNA marker technology

A comparison between two recently developed, PCR-based DNA marker technologies (amplified fragment length polymorphism, AFLP; inverse sequence-lagged repeat analysis, ISTR) was carried out in a group of 19 Vitis vinifera L. accessions, including 13 putative Sangiovese-related grapevines and 6 ''coloured'' ecotypes whose fruits are of importance for conferring intense red colour to the wine. A large amount of polymorphic DNA fragments was revealed by both molecular techniques: 8 different AFLP and 5 ISTR primer combinations generated 264 and 249 polymorphic markers, respectively. Similarity relationships among the accessions were described by cluster analysis. The AFLP analysis revealed the existence of a uniform group for the Sangiovese (SG) ecotypes showing a high degree of genetic relatedness for the members of this cultivar. Among the coloured ecotypes (CLR), variability was more evident. Only the so called Colorino americano ecotype significantly diverged from both groups. ISTR analysis confirmed the genetic dissimilarity of Colorino americano and the existence of the SG and CLR groups, but in addition detected a higher proportion of polymorphism among the Sangiovese accessions compared to AFLP analysis. Sangiovese forte and Saragiolo apparently differed from the other SG-related grapevines in agreement with AFLP results. It is possible that the observed genetic dissimilarity between Sangiovese forte, Saragiolo and other SG-related types could be interpreted by the putative polyclonal origin of many grapevine cultivars, a concept which is generally accepted by the grapevine research community. Both AFLP and ISTR appear to represent innovative, efficient and sensitive molecular tools for investigating genetic diversity among Vitis vinifera ecotypes and for the eventual identification of clones

Could be the swine responsible of transmission to the humans of Dientamoeba fragilis infection?

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miR-146a-5p impairs melanoma resistance to kinase inhibitors by targeting COX2 and regulating NFkB-mediated inflammatory mediators

BACKGROUND: Targeted therapy with BRAF and MEK inhibitors has improved the survival of patients with BRAF-mutated metastatic melanoma, but most patients relapse upon the onset of drug resistance induced by mechanisms including genetic and epigenetic events. Among the epigenetic alterations, microRNA perturbation is associated with the development of kinase inhibitor resistance. Here, we identified and studied the role of miR-146a-5p dysregulation in melanoma drug resistance.METHODS: The miR-146a-5p-regulated NFkB signaling network was identified in drug-resistant cell lines and melanoma tumor samples by expression profiling and knock-in and knock-out studies. A bioinformatic data analysis identified COX2 as a central gene regulated by miR-146a-5p and NFkB. The effects of miR-146a-5p/COX2 manipulation were studied in vitro in cell lines and with 3D cultures of treatment-resistant tumor explants from patients progressing during therapy.RESULTS: miR-146a-5p expression was inversely correlated with drug sensitivity and COX2 expression and was reduced in BRAF and MEK inhibitor-resistant melanoma cells and tissues. Forced miR-146a-5p expression reduced COX2 activity and significantly increased drug sensitivity by hampering prosurvival NFkB signaling, leading to reduced proliferation and enhanced apoptosis. Similar effects were obtained by inhibiting COX2 by celecoxib, a clinically approved COX2 inhibitor.CONCLUSIONS: Deregulation of the miR-146a-5p/COX2 axis occurs in the development of melanoma resistance to targeted drugs in melanoma patients. This finding reveals novel targets for more effective combination treatment. Video Abstract

How future surgery will benefit from SARS-COV-2-related measures: a SPIGC survey conveying the perspective of Italian surgeons

COVID-19 negatively affected surgical activity, but the potential benefits resulting from adopted measures remain unclear. The aim of this study was to evaluate the change in surgical activity and potential benefit from COVID-19 measures in perspective of Italian surgeons on behalf of SPIGC. A nationwide online survey on surgical practice before, during, and after COVID-19 pandemic was conducted in March-April 2022 (NCT:05323851). Effects of COVID-19 hospital-related measures on surgical patients' management and personal professional development across surgical specialties were explored. Data on demographics, pre-operative/peri-operative/post-operative management, and professional development were collected. Outcomes were matched with the corresponding volume. Four hundred and seventy-three respondents were included in final analysis across 14 surgical specialties. Since SARS-CoV-2 pandemic, application of telematic consultations (4.1% vs. 21.6%; p < 0.0001) and diagnostic evaluations (16.4% vs. 42.2%; p < 0.0001) increased. Elective surgical activities significantly reduced and surgeons opted more frequently for conservative management with a possible indication for elective (26.3% vs. 35.7%; p < 0.0001) or urgent (20.4% vs. 38.5%; p < 0.0001) surgery. All new COVID-related measures are perceived to be maintained in the future. Surgeons' personal education online increased from 12.6% (pre-COVID) to 86.6% (post-COVID; p < 0.0001). Online educational activities are considered a beneficial effect from COVID pandemic (56.4%). COVID-19 had a great impact on surgical specialties, with significant reduction of operation volume. However, some forced changes turned out to be benefits. Isolation measures pushed the use of telemedicine and telemetric devices for outpatient practice and favored communication for educational purposes and surgeon-patient/family communication. From the Italian surgeons' perspective, COVID-related measures will continue to influence future surgical clinical practice

Infected pancreatic necrosis: outcomes and clinical predictors of mortality. A post hoc analysis of the MANCTRA-1 international study

: The identification of high-risk patients in the early stages of infected pancreatic necrosis (IPN) is critical, because it could help the clinicians to adopt more effective management strategies. We conducted a post hoc analysis of the MANCTRA-1 international study to assess the association between clinical risk factors and mortality among adult patients with IPN. Univariable and multivariable logistic regression models were used to identify prognostic factors of mortality. We identified 247 consecutive patients with IPN hospitalised between January 2019 and December 2020. History of uncontrolled arterial hypertension (p = 0.032; 95% CI 1.135-15.882; aOR 4.245), qSOFA (p = 0.005; 95% CI 1.359-5.879; aOR 2.828), renal failure (p = 0.022; 95% CI 1.138-5.442; aOR 2.489), and haemodynamic failure (p = 0.018; 95% CI 1.184-5.978; aOR 2.661), were identified as independent predictors of mortality in IPN patients. Cholangitis (p = 0.003; 95% CI 1.598-9.930; aOR 3.983), abdominal compartment syndrome (p = 0.032; 95% CI 1.090-6.967; aOR 2.735), and gastrointestinal/intra-abdominal bleeding (p = 0.009; 95% CI 1.286-5.712; aOR 2.710) were independently associated with the risk of mortality. Upfront open surgical necrosectomy was strongly associated with the risk of mortality (p < 0.001; 95% CI 1.912-7.442; aOR 3.772), whereas endoscopic drainage of pancreatic necrosis (p = 0.018; 95% CI 0.138-0.834; aOR 0.339) and enteral nutrition (p = 0.003; 95% CI 0.143-0.716; aOR 0.320) were found as protective factors. Organ failure, acute cholangitis, and upfront open surgical necrosectomy were the most significant predictors of mortality. Our study confirmed that, even in a subgroup of particularly ill patients such as those with IPN, upfront open surgery should be avoided as much as possible. Study protocol registered in ClinicalTrials.Gov (I.D. Number NCT04747990)

Global disparities in surgeons’ workloads, academic engagement and rest periods: the on-calL shIft fOr geNEral SurgeonS (LIONESS) study

: The workload of general surgeons is multifaceted, encompassing not only surgical procedures but also a myriad of other responsibilities. From April to May 2023, we conducted a CHERRIES-compliant internet-based survey analyzing clinical practice, academic engagement, and post-on-call rest. The questionnaire featured six sections with 35 questions. Statistical analysis used Chi-square tests, ANOVA, and logistic regression (SPSS® v. 28). The survey received a total of 1.046 responses (65.4%). Over 78.0% of responders came from Europe, 65.1% came from a general surgery unit; 92.8% of European and 87.5% of North American respondents were involved in research, compared to 71.7% in Africa. Europe led in publishing research studies (6.6 ± 8.6 yearly). Teaching involvement was high in North America (100%) and Africa (91.7%). Surgeons reported an average of 6.7 ± 4.9 on-call shifts per month, with European and North American surgeons experiencing 6.5 ± 4.9 and 7.8 ± 4.1 on-calls monthly, respectively. African surgeons had the highest on-call frequency (8.7 ± 6.1). Post-on-call, only 35.1% of respondents received a day off. Europeans were most likely (40%) to have a day off, while African surgeons were least likely (6.7%). On the adjusted multivariable analysis HDI (Human Development Index) (aOR 1.993) hospital capacity > 400 beds (aOR 2.423), working in a specialty surgery unit (aOR 2.087), and making the on-call in-house (aOR 5.446), significantly predicted the likelihood of having a day off after an on-call shift. Our study revealed critical insights into the disparities in workload, access to research, and professional opportunities for surgeons across different continents, underscored by the HDI

Characterization of genetic biodiversity with Vitis vinifera L. Sangiovese and Colorino genotypes by AFLP and ISTR DNA marker technology

A comparison between two recently developed, PCR-based DNA marker technologies (amplified fragment length polymorphism, AFLP; inverse sequence-tagged repeat analysis, ISTR) was carried out in a group of 19 Vitis vinifera L. accessions, including 13 putative Sangiovese-related grapevines and 6 "coloured" ecotypes whose fruits are of importance for conferring intense red colour to the wine. A large amount of polymorphic DNA fragments was revealed by both molecular techniques: 8 different AFLP and 5 ISTR primer combinations generated 264 and 249 polymorphic markers, respectively. Similarity relationships among the accessions were described by cluster analysis. The AFLP analysis revealed the existence of a uniform group for the Sangiovese (SG) ecotypes showing a high degree of genetic relatedness for the members of this cultivar. Among the coloured ecotypes (CLR), variability was more evident. Only the so called Colorino americano ecotype significantly diverged from both groups. ISTR analysis confirmed the genetic dissimilarity of Colorino americano and the existence of the SG and CLR groups, but in addition detected a higher proportion of polymorphism among the Sangiovese accessions compared to AFLP analysis. Sangiovese forte and Saragiolo apparently differed from the other SG-related grapevines in agreement with AFLP results. It is possible that the observed genetic dissimilarity between Sangiovese forte, Saragiolo and other SG-related types could be interpreted by the putative polyclonal origin of many grapevine cultivars, a concept which is generally accepted by the grapevine research community. Both AFLP and ISTR appear to represent innovative, efficient and sensitive molecular tools for investigating genetic diversity among Vitis vinifera ecotypes and for the eventual identification of clones

Dientamoeba fragilis nella popolazione suina: indagini preliminari

Dientamoeba fragilis is a worldwide distributed protozoon which is now estimated responsible of enteric diseases in humans. A wide spectrum of intestinal symptoms has been described: diarrhoea (acute or prolonged), flatulence, abdominal pains, and not specific bowel syndrome; however, asymptomatic infection has been also reported. D. fragilis is still not well know because only the delicate trophozoite stage has been demonstrated in stool samples and definitive identification is based on permanent stains, usually not performed in ova and parasites (O&P) examinations, in Italy.D. fragilis is difficult to cultivate but molecular techniques, such as PCR, appear promising for a sure diagnosis of this protozoan infection.The epidemiology of dientamoebiasis is not clear too, and this paper presents preliminary results concerning the presence of D. fragilis in swine faeces in order to evaluate the possible role of pigs as reservoir of the parasite. So, 121 stool specimens of swine population were examined using Giemsa permanent stain and D. fragilis was observed in 53 stool specimens (43.8%). Other consideratioona are reported too