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Epithelial ion and fluid secretion determine the physiological functions of a broad range of organs, such as the lung, liver, or pancreas. The molecular mechanism of pancreatic ion secretion is challenging to investigate due to the limited access to functional human ductal epithelia. Patient-derived organoids may overcome these limitations, however direct accessibility of the apical membrane is not solved. In addition, due to the vectorial transport of ions and fluid the intraluminal pressure in the organoids is elevated, which may hinder the study of physiological processes. To overcome these, we developed an advanced culturing method for human pancreatic organoids based on the removal of the extracellular matrix that induced an apical-to-basal polarity switch also leading to reversed localization of proteins with polarized expression. The cells in the apical-out organoids had a cuboidal shape, whereas their resting intracellular Ca 2+ concentration was more consistent compared to the cells in the apical-in organoids. Using this advanced model, we demonstrated the expression and function of two novel ion channels, the Ca 2+ activated Cl - channel Anoctamin 1 (ANO1) and the epithelial Na + channel (ENaC), which were not considered in ductal cells yet. Finally, we showed that the available functional assays, such as forskolin-induced swelling, or intracellular Cl - measurement have improved dynamic range when performed with apical-out organoids. Taken together our data suggest that polarity-switched human pancreatic ductal organoids are suitable models to expand our toolset in basic and translational research., (© 2023. The Author(s).)

Background: Percutaneous transhepatic gallbladder drainage (PTGBD) plays an important role in the treatment of elderly patients and/or patients in poor health with acute cholecystitis (AC). The primary aim of this study is to determine how these factors influence the clinical outcome of PTGBD. Moreover, we assessed the timing and results of subsequent cholecystectomies., Patients and Methods: We retrospectively examined the results of 162 patients undergoing PTGBD between 2010 and 2020 (male-female ratio: 51.23% vs. 48.77%; mean age: 71.43 ± 13.22 years). Patient's performance status and intervention outcomes were assessed with clinical success rates (CSR) and in-hospital mortality. The conversion rate (CR) of possible urgent or delayed, elective laparoscopic cholecystectomies (LC) after PTGBD were analysed., Results: PTGBD was the definitive treatment in 42.18% of patients, while it was a bridging therapy prior to cholecystectomy (CCY) for the other patients. CSR was 87.97%, it was only 64.29% in grade III AC. In 9.87% of the cases, urgent LC was necessary after PTGBD, and its conversion rate was approximately equal to that of elective LC (18.18 vs. 17.46%, respectively, p = 0.2217). Overall, the post-PTGBD in-hospital mortality was 11.72%, while the same figure was 0% for grade I AC, 7.41% for grade II and 40.91% for grade III. Based on logistic regression analyses, in-hospital mortality (OR 6.07; CI 1.79-20.56), clinical progression (OR 7.62; CI 2.64-22.05) and the need for emergency CCY (OR 14.75; CI 3.07-70.81) were mostly determined by AC severity grade., Conclusion: PTGBD is an easy-to-perform intervention with promising clinical success rates in the treatment of acute cholecystitis. After PTGBD, the level of gallbladder inflammation played a decisive role in the course of AC. In a severe, grade III inflammation, we have to consider low CSR and high mortality., (© 2021. The Author(s).)

Background: The European Association of Endoscopic Surgery (EAES) fellowship programme was established in 2014, allowing nine surgeons annually to obtain experience and skills in minimally invasive surgery (MIS) from specialist centres across the Europe and United States. It aligns with the strategic focus of EAES Education and Training Committee on enabling Learning Mobility opportunities. To assess the impact of the programme, a survey was conducted aiming to evaluate the experience and impact of the programme and receive feedback for improvements., Methods: A survey using a 5-point Likert scale was used to evaluate clinical, education and research experience. The impact on acquisition of new technical skills, change in clinical practice and ongoing collaboration with the host institute was assessed. The fellows selected between 2014 and 2018 were included. Ratings were analysed in percentage; thematic analysis was applied to the free-text feedbacks using qualitative analysis., Results: All the fellows had good access to observing in operating theatres and 70.6% were able to assist. 91.2% participated in educational activities and 23.5% were able to contribute through teaching. 44.1% participated in research activities and 41.2% became an author/co-author of a publication from the host. 97.1% of fellows stated that their operative competency had increased, 94.3% gained new surgical skills and 85.7% was able to introduce new techniques in their hospitals. 74.29% agreed that the clinical experience led to a change in their practices. The most commonly suggested improvements were setting realistic target in clinical and research areas, increasing fellowship duration, and maximising theatre assisting opportunities. Nevertheless, 100% of fellows would recommend the fellowship to their peers., Conclusion: EAES fellowship programme has shown a positive impact on acquiring and adopting new MIS techniques. To further refine the programme, an individualised approach should be adopted to set achievable learning objectives in clinical skills, education and research., (© 2021. The Author(s).)

Background: The Alvarado score is a clinical scoring system used in the diagnosis of acute appendicitis. This study aimed to compare the reliability of the Alvarado score and clinical judgment and to refine the score to make it easier to use., Methods: In this prospective, randomized study, patients presenting at the authors' outpatient department with suspected appendicitis during a 1-year period were assigned in weekly alternation to either group A or group B. The group A patients were treated on the basis of their Alvarado score, and the group B patients underwent treatment based on clinical judgment. The correctness of the methods was assessed by evaluation of the final histology. Statistical comparison of the data was performed using SPSS 20., Results: The study investigated 269 patients (131 in group A and 138 in group B). The groups were homogeneous in terms of mean age, gender, body mass index, and American Society of Anesthesiologists score. The number of negative appendectomies was 12 (9.16%) in group A versus 5 (3.6%) in group B (p = 0.063). The clinical judgment had better specificity and sensitivity than the Alvarado score. For that reason, the specificity of the Alvarado score was refined using statistical methods, with weighting of certain clinical data and inclusion of new ones (e.g., ultrasound investigation). Consequently, the area under the curve by receiver operating characteristic analysis gradually increased, and the Alvarado score became more accurate., Conclusion: The study findings showed clinical judgment to be more reliable in the diagnosis of acute appendicitis than the Alvarado score, but the score is a useful diagnostic aid, especially for young colleagues. The use of the new scoring system has become easier. It includes fewer criteria as well as an important and sensitive predictor: the ultrasound investigation.

Myofibroblasts play central roles in wound healing, deposition of the extracellular matrix and epithelial function. Their functions depend on migration and proliferation within the subepithelial matrix, which results in accelerated cellular metabolism. Upregulated metabolic pathways generate protons which need to be excreted to maintain intracellular pH (pH(i)). We isolated human gastric myofibroblasts (HGMs) from surgical specimens of five patients. Then we characterized, for the first time, the expression and functional activities of the Na(+)/H(+) exchanger (NHE) isoforms 1, 2 and 3, and the functional activities of the Na(+)/HCO(3)(-) cotransporter (NBC) and the anion exchanger (AE) in cultured HGMs using microfluorimetry, immunocytochemistry, reverse transcription polymerase chain reaction and immunoblot analysis. We showed that NHE1-3, NBC and AE activities are present in HGMs and that NHE1 is the most active of the NHEs. In scratch wound assays we also demonstrated (using the selective NHE inhibitor HOE-642) that carbachol and insulin like growth factor II (IGF-II) partly stimulate migration of HGMs in a NHE1-dependent manner. EdU incorporation assays revealed that IGF-II induces proliferation of HGMs which is inhibited by HOE-642. The results indicate that NHE1 is necessary for IGF-II-induced proliferation response of HGMs. Overall, we have characterized the pH(i) regulatory mechanisms of HGMs. In addition, we demonstrated that NHE1 activity contributes to both IGF-II- and carbachol-stimulated migration and that it is obligatory for IGF-II-induced proliferation of HGMs.

Background: This retrospective single-institution study presents a successful treatment strategy for Boerhaave's syndrome., Methods: During 1995-2008, 15 patients with spontaneous esophageal perforation were treated. Patients were grouped according to time from symptoms to referral (early, <24 h; late, >24 h). In group I (early, n = 8 patients) treatment comprised primary surgical esophageal repair in seven cases and endoscopic clipping in one case. In group II (late, n = 7 patients) treatment comprised esophagectomy without primary reconstruction (4 cases) or controlled esophagocutaneous fistula (3 cases). Measures of outcome included age (years), delay to diagnosis (h), severe sepsis on admission, mortality, and hospital and intensive care unit (ICU) stay., Results: The overall hospital mortality rate was 6.6% (1/15), being 0% (0/8) in group I and 14.2% (1/7) in group II. Patient age (49.6 vs. 68.6 years, P < 0.0001), delay to diagnosis (17.75 vs. 69 h, P < 0.0001), severe sepsis on admission (0 vs. 4, P = 0.0256), and ICU stay (4 vs. 14 days, P = 0.006) were all greater in group II., Conclusions: Early diagnosis and carefully selected therapeutic tactics can reduce the mortality rate of Boerhaave's syndrome to an acceptably low level. Methods of organ preservation and minimally invasive techniques can be applied successfully in the treatment.

Protoplasts isolated from cultured cells of albino carrot (Daucus carota) and normal green D. capillifolius were fused by polyethylene glycol. Selection of somatic hybrid plants was based on the restoration of photosynthetic function in hybrids. Green plantlets selected from embryo cultures were characterized on the basis of leaf morphology. The interspecific protoplast fusion resulted in green plants with leaves which were intermediate between those of the parents. The somatic hybrids between orange rooted carrot variety and D. capillifolius with long white roots produced long, white and fleshy roots. The cytological analysis of parasexual hybrids revealed that the chromosome number ranged from 34 to 54. The most frequent chromosome number was 2n = 36. Hybrids were also found with 34 and 35 chromosomes. The somatic hybrid showed the same isoenzyme pattern of leaf peroxidase as D. carota.

Mitochondrial DNA (mtDNA) restriction patterns were studied in mutant, cybrid and somatic hybrid plants regenerated from Nicotiana protoplasts.It has been shown that neither components of the culture media used for protoplast culture and plant regeneration, nor the antibiotics streptomycin and lincomycin used for the mutant selection induced alterations in the mtDNA. No rearrangements were detected in the mtDNA of plants derived from homoplasmic fusions where the mtDNA of the parents was identical as judged by mtDNA restriction patterns.There were rearrangements, however, in the mtDNA of each of the cybrid plants derived from heteroplasmic fusions. Restriction patterns generated by BamHI and SalI restriction endonucleases were different from those of both parents, and were composed of parental and non-parental fragments.