SELECTION OF METHODS OF COMBINED GERNIOPLASTICS EXTENSIVE MEDIAN HERNIAS TAKING INTO ACCOUNT THE DYSPLASIA OF CONNECTING TISSUE

For the purpose of improvement of results of treatment of extensive median hernias in the choice of ways of the combined hernioplasty at a stage of an electromyography ball assessment of the stigmata of a dysplasia of a connecting tissue, influence of a mesenchymal failure on contractility of abdominal muscles and data of program diagnostics of a collagen found at survey in microscopic preparations of a skin and aponeurosis at 95 surgical patients is introduced. In group 25 (26,4%) of patients with clinically significant level of a dysplasia depression of electroactivity of rectus muscles for 24,7% and the lateral group of the abdominal muscles - for 22,8% is revealed. The microscopy of sites of an aponeurosis among them taped depression of density of laying of a collagen to 31,7% and augmentation of intensity of its staining twice. As a result of the undertaken improvement the way of surgical treatment of median hernias of the extensive sizes which use in clinical practice allows to reduce a share of a dysplasia of a connecting tissue among the reasons of a recurrence of a disease is developed

Towards a methodology for the engineering of event-driven process applications

Successful applications of the Internet of Things such as smart cities, smart logistics, and predictive maintenance, build on observing and analyzing business-related objects in the real world for business process execution and monitoring. In this context, complex event processing is increasingly used to integrate events from sensors with events stemming from business process management systems. This paper describes a methodology to combine the areas and engineer an event-driven logistics processes application. Thereby, we describe the requirements, use cases and lessons learned to design and implement such an architecture

Virtual patients design and its effect on clinical reasoning and student experience : a protocol for a randomised factorial multi-centre study

Background Virtual Patients (VPs) are web-based representations of realistic clinical cases. They are proposed as being an optimal method for teaching clinical reasoning skills. International standards exist which define precisely what constitutes a VP. There are multiple design possibilities for VPs, however there is little formal evidence to support individual design features. The purpose of this trial is to explore the effect of two different potentially important design features on clinical reasoning skills and the student experience. These are the branching case pathways (present or absent) and structured clinical reasoning feedback (present or absent). Methods/Design This is a multi-centre randomised 2x2 factorial design study evaluating two independent variables of VP design, branching (present or absent), and structured clinical reasoning feedback (present or absent).The study will be carried out in medical student volunteers in one year group from three university medical schools in the United Kingdom, Warwick, Keele and Birmingham. There are four core musculoskeletal topics. Each case can be designed in four different ways, equating to 16 VPs required for the research. Students will be randomised to four groups, completing the four VP topics in the same order, but with each group exposed to a different VP design sequentially. All students will be exposed to the four designs. Primary outcomes are performance for each case design in a standardized fifteen item clinical reasoning assessment, integrated into each VP, which is identical for each topic. Additionally a 15-item self-reported evaluation is completed for each VP, based on a widely used EViP tool. Student patterns of use of the VPs will be recorded. In one centre, formative clinical and examination performance will be recorded, along with a self reported pre and post-intervention reasoning score, the DTI. Our power calculations indicate a sample size of 112 is required for both primary outcomes

Epstein–Barr virus LMP1 oncogene polymorphism in tatar and slavic populations in Russian Federation impacting on some malignant tumours

Objective: To compare genetic structure of the main Epstein–Barr virus (EBV) oncogene, latent membrane protein 1 (LMP1), in EBV strains circulating in two genetically distinct ethnic populations in Russian Federation, Tatars and Slavs, as well as assess an impact of diverse LMP1 variants on incidence and mortality rate for some malignant tumors partially associated with EBV infection. Materials and methods. Oral washing samples were collected from 60 ethnic Kazan Tatars and 65 ethnic Moscow Slavics. Carboxy-terminal nucleotide sequences (41 and 40 sequences, respectively) derived from hypervariable LMP1 gene region were amplified from EBV DNA samples. Next, final nucleotide sequences were translated into amino acid sequences and analyzed according to classification by Edwards et al. Results. Analysis of 41 and 40 LMP1 samples obtained from ethnic Kasan Tatars and ethnic Moscow Slavics, respectively, revealed significant difference in relevant amino acid structures. In particular, all LMP1 samples derived from Moscow Slavics were found to belong to the four protein variants: B95.8/A, Med–, China1 and NC. Among them, low-transforming variant B95.8/A was dominant (82.5%). In contrast, solely 21 out of 41 LMP1 samples derived from ethnic Tatars were classified as B95.8/A, Med– and China1 variants. Importantly, the percentage of low-transforming B95.8/A variant among ethnic Tatar samples was significantly lower compared to that one found in Moscow Slavics (29.3% vs. 82.5%). On the other hand, seven (17.1%) out of 20 other samples formed a unique protein mono group characterized by LMP1 amino acid sequence differed from that one available in the GenBank database. Such group of variants was designated as LMP1-TatK. The remaining 13 samples (31.7%) did not match either protein variants, thereby forming the “beyond classification” (LMP1-TatBC) group. Conclusion. The data obtained suggest that various LMP1 variants exist in EBV strains persisting in ethnic Tatrs and ethnic Slavics examined in Russian Federation. It was also found that EBV strains isolated from ethnic Tatars contained a unique LMP1 gene variant encoding protein LMP1-TatK lacked in EBV strains derived from ethnic Moscow Slavics. Taking into account the genealogy of Tatars, it cannot be ruled out that EBV strain bearing LMP1-TatK variant represented ethnically specific EBV strain that might circulate many centuries ago among their historical human predecessors called Mongol-Tatar tribes. In addition, it was shown that the LMP1 variants in EBV strains isolated from ethnic Kazan Tatars and ethnic Moscow Slavics did not affect the incidence and mortality of different forms of cancer consisting of EBV-associated cases

Выявление мутаций в «горячих» участках генома: ампликоны-«шпильки» в методе плавления ДНК

Polymerase chain reaction (PCR) followed by DNA melting analysis with TaqMan probes effectively reveals mutations in the human genome “hot” spots. The necessity to carry out PCR in the asymmetric variant causes, however, a number of restrictions of this method: 1) an inability of quantitative estimates of gene copy numbers; 2) the need for 2 independent PCR tests for detection of mutations in both complementary strands of an amplicon (this approach improves reliability and sensitivity of the analysis); 3) the complication of PCR design and decrease in efficiency of amplification. Overcoming these restrictions was possible by means of symmetric PCR with primers containing the specific and universal sequences: the single-stranded “hairpins” (sense and antisense) are not capable to anneal with each other, but they can hybridize independently with 2 TaqMan probes present in the reaction mixture. The proposed approach allows quantitative and qualitative characterization of a DNA sample (the copy number estimates as well as mutation scanning of both complementary amplicon strands).Амплификация с последующим плавлением ДНК с зондами TaqMan эффективно выявляет мутации в «горячих» участках генома. Однако необходимость проводить полимеразную цепную реакцию (ПЦР) в асимметричном варианте обусловливает ряд ограничений этого метода: 1) невозможность количественного анализа из‑за снижения эффективности амплификации; 2) необходимость выполнения 2 независимых ПЦР для выявления мутаций в комплементарных нитях ампликона; 3) усложнение дизайна ПЦР. Преодоление этих ограничений оказалось возможным при использовании в симметричной ПЦР комбинированных праймеров, состоящих из универсальной и специфической последовательностей: образующиеся в результате однонитевые «шпилечные» (hairpin) ампликоны (sense и antisense) не способны ренатурировать друг с другом, но независимо гибридизуются с присутствующими в среде зондами TaqMan (antisense и sense соответственно). Разработанный способ позволяет в одном тесте получить количественные (число копий) и качественные (наличие мутаций в обеих нитях ампликона) характеристики исследуемого участка ДНК

Serum Lipoprotein(a) and Bioprosthetic Aortic Valve Degeneration

AIMS: Bioprosthetic aortic valve degeneration demonstrates pathological similarities to aortic stenosis. Lipoprotein(a) [Lp(a)] is a well-recognized risk factor for incident aortic stenosis and disease progression. The aim of this study is to investigate whether serum Lp(a) concentrations are associated with bioprosthetic aortic valve degeneration. METHODS AND RESULTS: In a post hoc analysis of a prospective multimodality imaging study (NCT02304276), serum Lp(a) concentrations, echocardiography, contrast-enhanced computed tomography (CT) angiography, and 18F-sodium fluoride (18F-NaF) positron emission tomography (PET) were assessed in patients with bioprosthetic aortic valves. Patients were also followed up for 2 years with serial echocardiography. Serum Lp(a) concentrations [median 19.9 (8.4-76.4) mg/dL] were available in 97 participants (mean age 75 ± 7 years, 54% men). There were no baseline differences across the tertiles of serum Lp(a) concentrations for disease severity assessed by echocardiography [median peak aortic valve velocity: highest tertile 2.5 (2.3-2.9) m/s vs. lower tertiles 2.7 (2.4-3.0) m/s, P = 0.204], or valve degeneration on CT angiography (highest tertile n = 8 vs. lower tertiles n = 12, P = 0.552) and 18F-NaF PET (median tissue-to-background ratio: highest tertile 1.13 (1.05-1.41) vs. lower tertiles 1.17 (1.06-1.53), P = 0.889]. After 2 years of follow-up, there were no differences in annualized change in bioprosthetic hemodynamic progression [change in peak aortic valve velocity: highest tertile [0.0 (-0.1-0.2) m/s/year vs. lower tertiles 0.1 (0.0-0.2) m/s/year, P = 0.528] or the development of structural valve degeneration. CONCLUSION: Serum lipoprotein(a) concentrations do not appear to be a major determinant or mediator of bioprosthetic aortic valve degeneration

Высокочувствительное сканирование генных мутаций: зонды TaqMan как блокирующие агенты

DNA Melting Analysis is very effective in clinical DNA diagnostics: it is simple to perform, high throughput, labor-, time- and cost-effective and is implemented in the “closed tube” format minimizing the risk of samples cross-contamination. Although more sensitive than sequencing by Sanger (mutant allele detection limit is ~5 and ~15 % respectively), it, however, is inferior in this respect to some other, more laborious and expensive methods (in particular, ddPCR (digital droplet PCR)). Using the BRAF gene as a prototype, we developed the original version of the DNA melting analysis, based on the ability of TaqMan probes to hamper the primer extension reaction by Taq-polymerase. It is found that the weaker blocking effect on the mutant template, which is due to the mismatch in the probe-DNA heteroduplex, permits enriched amplification of the mutant allele and provides a significant (10-fold or more) increase in sensitivity of mutation scanning.Метод плавления ДНК весьма эффективен в клинической генодиагностике, прост в исполнении, производителен, экономичен и, кроме того, реализуется в «закрытом формате», сводящем к минимуму затраты времени, труда и риск перекрестного загрязнения образцов. Данный метод более чувствительный, чем секвенирование по Сэнгеру (предел обнаружения мутантных аллелей ~5 и ~15 % соответственно), однако уступает в этом отношении другим, более трудоемким и дорогим методам (в частности, капельной цифровой полимеразной цепной реакции (digital droplet PCR)). На гене BRAF (как прототипе) мы разработали оригинальный вариант метода плавления ДНК, основанный на способности зондов TaqMan затруднять движение Taq-полимеразы по матрице. Установлено, что эффект блокирования слабее выражен на мутантной матрице из-за присутствия в дуплексе зонд-ДНК неспаренного основания. Предложен протокол полимеразной цепной реакции, дискриминирующий амплификацию мутантных и нормальных аллелей и обеспечивающий существенное (10-кратное и более) повышение чувствительности мутационного сканирования

Rapid Qualitative Urinary Tract Infection Pathogen Identification by SeptiFast® Real-Time PCR

Background Urinary tract infections (UTI) are frequent in outpatients. Fast pathogen identification is mandatory for shortening the time of discomfort and preventing serious complications. Urine culture needs up to 48 hours until pathogen identification. Consequently, the initial antibiotic regimen is empirical. Aim To evaluate the feasibility of qualitative urine pathogen identification by a commercially available real-time PCR blood pathogen test (SeptiFast®) and to compare the results with dipslide and microbiological culture. Design of study Pilot study with prospectively collected urine samples. Setting University hospital. Methods 82 prospectively collected urine samples from 81 patients with suspected UTI were included. Dipslide urine culture was followed by microbiological pathogen identification in dipslide positive samples. In parallel, qualitative DNA based pathogen identification (SeptiFast®) was performed in all samples. Results 61 samples were SeptiFast® positive, whereas 67 samples were dipslide culture positive. The inter-methodological concordance of positive and negative findings in the gram+, gram- and fungi sector was 371/410 (90%), 477/492 (97%) and 238/246 (97%), respectively. Sensitivity and specificity of the SeptiFast® test for the detection of an infection was 0.82 and 0.60, respectively. SeptiFast® pathogen identifications were available at least 43 hours prior to culture results. Conclusion The SeptiFast® platform identified bacterial DNA in urine specimens considerably faster compared to conventional culture. For UTI diagnosis sensitivity and specificity is limited by its present qualitative setup which does not allow pathogen quantification. Future quantitative assays may hold promise for PCR based UTI pathogen identification as a supplementation of conventional culture methods

Right ventricle to pulmonary artery coupling after transcatheter aortic valve implantation—Determinant factors and prognostic impact

IntroductionRight ventricular (RV) dysfunction and pulmonary hypertension (PH) have been previously associated with unfavorable outcomes in patients with severe aortic stenosis (AS) undergoing transcatheter aortic valve implantation (TAVI), but little is known about the effect of right ventricle (RV) to pulmonary artery (PA) coupling. Our study aimed to evaluate the determinant factors and the prognostic value of RV-PA coupling in patients undergoing TAVI.MethodsOne hundred sixty consecutive patients with severe AS were prospectively enrolled, between September 2018 and May 2020. They underwent a comprehensive echocardiogram before and 30 days after TAVI, including speckle tracking echocardiography (STE) for myocardial deformation analysis of the left ventricle (LV), left atrium (LA), and RV function. Complete data on myocardial deformation was available in 132 patients (76.6 ± 7.5 years, 52.5% men) who formed the final study population. The ratio of RV free wall longitudinal strain (RV-FWLS) to PA systolic pressure (PASP) was used as an estimate of RV-PA coupling. Patients were analyzed according to baseline RV-FWLS/PASP cut-off point, determined through time-dependent ROC curve analysis, as follows: normal RV-PA coupling group (RV-FWLS/PASP ≥0.63, n = 65) and impaired RV-PA coupling group (RV-FWLS/PASP < 0.63, n = 67).ResultsA significant improvement of RV-PA coupling was observed early after TAVI (0.75 ± 0.3 vs. 0.64 ± 0.3 before TAVI, p < 0.001), mainly due to PASP decrease (p < 0.001). LA global longitudinal strain (LA-GLS) is an independent predictor of RV-PA coupling impairment before and after TAVI (OR = 0.837, p < 0.001, OR = 0.848, p < 0.001, respectively), while RV diameter is an independent predictor of persistent RV-PA coupling impairment after TAVI (OR = 1.174, p = 0.002). Impaired RV-PA coupling was associated with a worse survival rate (66.3% vs. 94.9%, p-value < 0.001) and emerged as an independent predictor of mortality (HR = 5.97, CI = 1.44–24.8, p = 0.014) and of the composite endpoint of death and rehospitalization (HR = 4.14, CI = 1.37–12.5, p = 0.012).ConclusionOur results confirm that relief of aortic valve obstruction has beneficial effects on the baseline RV-PA coupling, and they occur early after TAVI. Despite significant improvement in LV, LA, and RV function after TAVI, RV-PA coupling remains impaired in some patients, it is mainly related to persistent pulmonary hypertension and is associated with adverse outcomes