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Changes in intracellular pH (pHi) reflect metabolic states of cancer cells during tumor growth and dissemination. Therefore, monitoring of pHi is essential for understanding the metabolic mechanisms that support cancer progression. Genetically encoded fluorescent pH sensors have become irreplaceable tools for real-time tracking pH in particular subcellular compartments of living cells. However, ratiometric readout of most of the pH probes is poorly suitable to measure pH in thick samples ex vivo or tissues in vivo including solid tumors. Fluorescence lifetime imaging (FLIM) is a promising alternative to the conventional fluorescent microscopy. Here, we present a quantitative approach to map pHi in cancer cells and tumors in vivo, relying on fluorescence lifetime of a genetically encoded pH sensor SypHerRed. We demonstrate the utility of SypHerRed in visualizing pHi in cancer cell culture and in mouse tumor xenografts using fluorescence lifetime imaging microscopy and macroscopy. For the first time to our knowledge, the absolute pHi value is obtained for tumors in vivo by an optical technique. In addition, we demonstrate the possibility of simultaneous detection of pHi and endogenous fluorescence of metabolic cofactor NADH, which provides a complementary insight into metabolic aspects of cancer. Fluorescence lifetime-based readout and red-shifted spectra make pH sensor SypHerRed a promising instrument for multiparameter in vivo imaging applications.

Glofitamab, a novel CD20xCD3, T-cell–engaging bispecific antibody, exhibited single-agent activity in Study NP30179, a first-in-human, phase 1 trial in relapsed/refractory B-cell non-Hodgkin lymphoma. Preclinical studies showed that glofitamab leads to T-cell activation, proliferation, and tumor cell killing upon binding to CD20 on malignant cells. Here, we provide evidence of glofitamab’s clinical activity, including pharmacodynamic profile, mode of action, and factors associated with clinical response, by evaluating biomarkers in patient samples from the dose-escalation part of this trial. Patients enrolled in Study NP30179 received single-dose obinutuzumab pretreatment (1000 mg) 7 days before IV glofitamab (5 µg-25 mg). Glofitamab treatment lasted ≤12 cycles once every 2 or 3 weeks. Blood samples were collected at predefined time points per the clinical protocol; T-cell populations were evaluated centrally by flow cytometry, and cytokine profiles were analyzed. Immunohistochemical and genomic biomarker analyses were performed on tumor biopsy samples. Pharmacodynamic modulation was observed with glofitamab treatment, including dose-dependent induction of cytokines, and T-cell margination, proliferation, and activation in peripheral blood. Gene expression analysis of pretreatment tumor biopsy samples indicated that tumor cell intrinsic factors such as TP53 signaling are associated with resistance to glofitamab, but they may also be interlinked with a diminished effector T-cell profile in resistant tumors and thus represent a poor prognostic factor per se. This integrative biomarker data analysis provides clinical evidence regarding glofitamab’s mode of action, supports optimal biological dose selection, and will further guide clinical development. This trial was registered at www.clinicaltrials.gov as #NCT03075696.

Genetically encoded pH-sensors are the promising instrument for intracellular pH (pHi) registration. In tumor tissue the reversed pH gradient is known to be the important hallmark of cancer and regulator of tumor response on chemotherapy. However the effect of chemotherapeutic drugs on the pHi of tumor cells is largely unknown. Here we using genetically encoded ratiometric pH-sensor SypHer2 were able to monitor pHi in vitro in cell monolayer and tumor spheroids and in vivo in tumor xenografts. In tumor cell monolayer different pHi dynamic was revealed in the dying cell and division-arrested surviving cells. The treatment effect of taxol varied in monolayer and tumor spheroids and pHi changes were able to reflect these difference. The tend to pHi decrease in respect to taxol in vivo matched with results obtained for the cell monolayer. Also in both cases the cell cycle-arrest was the main treatment effect in contrast to tumor spheroid, where the cell death was the primary result. These findings elucidate the significance of pHi in the mechanisms of taxol action on cervical cancer cells and will be valuable for development of new approaches for cancer treatment.

Aim To study features of diagnosis and treatment of acute myocardial infarction (AMI) in Russian hospitals, results of the treatment, and early and late outcomes (6 and 12 months after AMI diagnosis); to evaluate the consistence of the treatment with clinical guidelines; and to evaluate patients’ compliance with the treatment.Material and methods The program was designed for 3 years, including 24 months for recruitment of patients to the study. The study will include 10, 000 patients hospitalized with a confirmed diagnosis (I21 according to ICD-10) of ST segment elevation acute myocardial infarction (MI) (STEMI) or non-ST segment elevation MI (NSTEMI) based on criteria of the European Society of Cardiology Guidelines on Forth Universal Definition of Myocardial Infarction (2018). The follow-up period was divided into three stages: observation during the stay in the hospital and at 6 and 12 months following inclusion into the registry. The primary endpoint included cardiac death, nonfatal MI during the hospitalization and after one-year follow-up. Secondary endpoints were 6-months and one-year incidence of repeated MI, heart failure, ischemic stroke, clinically significant hemorrhage, unscheduled revascularization after discharge from the hospital, and the proportion of patients who continue on statins, antiplatelet drugs, and drugs of other groups for 6 months and 1 year.Results The inclusion of patients into the registry started in 2020 and will continue for 24 months. By the time of the article publication (June, 2021), more than 2,000 patients will be included.Conclusion REGION-MI (Russian rEGIstry Of acute myocardial iNfarction) is a multicenter, retrospective and prospective observational cohort study that excludes any interference with the clinical practice. Results of the registry will help to analyze a real picture of medical care provided to patients with myocardial infarction and to schedule ways to improve the situation.

A clinical case of incorrect verification of the angiosarcoma of the ethmoidal labyrinth and frontal sinus in a 36-year-old man is presented. A feature of this case is the disregard of additional research methods without taking into account the unilateralism of the pathological process, which led to the wrong tactics of surgical treatment and the development of a relapse of the disease complicated by nasal liquorrhea. A thorough examination of the patient and an analysis of a series of computer tomograms of the paranasal sinuses suggested a diagnosis of malignancy. The patient underwent neoplasm removal with fistula plastic, and histological confirmation of the presumptive diagnosis was performed.Представлен клинический случай некорректной верификации ангиосаркомы решетчатого лабиринта и лобной пазухи у 36-летнего мужчины. Особенностью является игнорирование дополнительных методов исследования, одностороннего характера патологического процесса, приведшее к ошибочной тактике хирургического лечения и развитию рецидива заболевания, осложнившегося назальной ликвореей. Тщательный осмотр пациента, анализ серии компьютерных томограмм околоносовых пазух позволил предположить диагноз злокачественного новообразования. Пациенту проведено удаление новообразования с пластикой ликворного свища и гистологическим подтверждением предполагаемого диагноза.

Non-invasive radionuclide imaging of human epidermal growth factor receptor type 2 (HER2) expression in breast, gastroesophageal, and ovarian cancers may stratify patients for treatment using HER2-targeted therapeutics. Designed ankyrin repeat proteins (DARPins) are a promising type of targeting probe for radionuclide imaging. In clinical studies, the DARPin [

CD133 is an extensively studied marker of the most malignant tumor cell population, designated as cancer stem cells (CSCs). However, the function of this glycoprotein and its involvement in cell regulatory cascades are still poorly understood. Here we show a positive correlation between the level of CD133 plasma membrane expression and the proliferative activity of cells of the Caco-2, HT-29, and HUH7 cancer cell lines. Despite a substantial difference in the proliferative activities of cell populations with different levels of CD133 expression, transcriptomic and proteomic profiling revealed only minor distinctions between them. Nonetheless, a further in silico assessment of the differentially expressed transcripts and proteins revealed 16 proteins that could be involved in the regulation of CD133 expression; these were assigned ranks reflecting the apparent extent of their involvement. Among them, the TRIM28 transcription factor had the highest rank. The prominent role of TRIM28 in CD133 expression modulation was confirmed experimentally in the Caco2 cell line clones: the knockout, though not the knockdown, of the TRIM28 gene downregulated CD133. These results for the first time highlight an important role of the TRIM28 transcription factor in the regulation of CD133-associated cancer cell heterogeneity.

Background Coronavirus disease 2019 (COVID-19) was declared a pandemic by the WHO on 11 March 2020 and global surgical practice was compromised. This Commission aimed to document and reflect on the changes seen in the surgical environment during the pandemic, by reviewing colleagues’ experiences and published evidence. Methods In late 2020, BJS contacted colleagues across the global surgical community and asked them to describe how severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) had affected their practice. In addition to this, the Commission undertook a literature review on the impact of COVID-19 on surgery and perioperative care. A thematic analysis was performed to identify the issues most frequently encountered by the correspondents, as well as the solutions and ideas suggested to address them. Results BJS received communications for this Commission from leading clinicians and academics across a variety of surgical specialties in every inhabited continent. The responses from all over the world provided insights into multiple facets of surgical practice from a governmental level to individual clinical practice and training. Conclusion The COVID-19 pandemic has uncovered a variety of problems in healthcare systems, including negative impacts on surgical practice. Global surgical multidisciplinary teams are working collaboratively to address research questions about the future of surgery in the post-COVID-19 era. The COVID-19 pandemic is severely damaging surgical training. The establishment of a multidisciplinary ethics committee should be encouraged at all surgical oncology centres. Innovative leadership and collaboration is vital in the post-COVID-19 era.

Cardiovascular complications including arrhythmias and cardiac conduction disorders are one of the main reasons of high mortality rate in acromegaly, while they have not been well explored.To estimate arrhythmias frequency in acromegaly, identify risk factors leading to the development of arrhythmia and cardiac conduction disorder, to determine the role of cardiac MRI in detecting structural and functional changes.A single-center prospective cohort study, which included 461 patients (151 men and 310 women) with acromegaly, was conducted. All the patients underwent a standard medical examination, including hormonal blood test, electrocardiogram, echocardiography, electrocardiogram daily monitoring. 18 patients with arrhythmias (11 men and 7 women) had cardiac MRI with gadolinium-based contrast.The results of our research show high frequency of arrhythmias and cardiac conduction disorders in patients with acromegaly 42%. Most frequent kinds of arrhythmias and cardiac conduction disorders were sinus bradycardia 19.1% of the cases and conduction disorders of bundle branch blocks 14.5%. Men were more likely to suffer from arrhythmias and cardiac conduction disorders than women (54.2% and 37.4%, respectively,p=0.0005). Not acromegaly activity but duration of the disease was a main risk factor of arrhythmias and cardiac conduction disorders. Patients with arrhythmias had a long anamnesis of acromegaly (10 and 7 years, respectively, p=0.04). Meanwhile, cardiac conduction disorders were commonly observed in the patients who were treated with somatostatin analogs comparing to the patients who didnt undergo this therapy (50% and 38.6% respectively,p=0.004). We showed that 61% of patients with acromegaly and cardiac conduction disorders who underwent magnetic resonance imaging (MRI) had the signs of myocardial fibrosis. The value of the ejection fraction of the left ventricle according to MRI was higher than with echocardiography (p=0.04).Arrhythmias and cardiac conduction disorders are often observed in patients with acromegaly even with remission of the disease. High risk group need careful diagnostic and monitoring approaches. Cardiac MRI is the gold standard for visualization of structural and morphological changes in the heart. Use of cardiac MRI in acromegalic patients expands our understanding of arrhythmias and cardiac conduction disorders in this disease. There are no specific laboratory markers of diffuse myocardial fibrosis, and the role of myocardial fibrosis in the occurrence of cardiac arrhythmias and conduction disorders needs further studying.Сердечно-сосудистые осложнения, включая нарушение ритма и проводимости сердца (НРПС), являются одной из основных причин высокой летальности при акромегалии, однако в настоящее время недостаточно изучены. Цель.Оценить частоту аритмий у пациентов с акромегалией, выявить факторы риска, способствующие развитию НРПС, определить роль магнитно-резонансной томографии (МРТ) сердца в выявлении структурных и функциональных изменений. Материалы и методы.Проведено одноцентровое проспективное когортное исследование, в которое включен 461 пациент (151 мужчина и 310 женщин) с акромегалией. Всем пациентам проведено стандартное медицинское обследование, включающее гормональное исследование крови, регистрацию электрокардиограммы (ЭКГ), эхокардиографию, холтеровское мониторирование ЭКГ. МРТ сердца с контрастированием гадолинием выполнено 18 пациентам с НРПС (11 мужчин и 7 женщин). Результаты.Результаты нашего исследования демонстрируют высокую встречаемость НРПС среди пациентов с акромегалией 42%. Наиболее частыми видами являлись синусовая брадикардия 19,1% случаев, а также нарушения проводимости по ножкам пучка Гиса 14,5%. Мужчины чаще страдали НРПС, чем женщины (54,2 и 37,4% соответственно,р=0,0005). Мы показали, что развитие НРПС зависит, в первую очередь, от длительности акромегалии, в то время как активность заболевания не оказывает значимого влияния. Анамнез акромегалии у пациентов с НРПС и без них составил 10 и 7 лет соответственно (p=0,04). Кроме того, важным фактором риска оказалась терапия аналогами соматостатина. Доля пациентов, получающих данный вид лечения, составила 50% с НРПС и 38,6% среди пациентов без НРПС (р=0,004). Мы продемонстрировали, что 61% пациентов с акромегалией и НРПС, которым провели МРТ сердца, имели признаки фиброза миокарда. При этом значение фракции выброса левого желудочка по данным МРТ оказалось статистически выше, чем при эхокардиографии (p=0,04). Заключение.НРПС часто встречаются у пациентов с акромегалией, несмотря на достижение ремиссии заболевания. Группа высокого риска требует более тщательной диагностики и мониторинга. МРТ сердца является золотым стандартом визуализации структурно-морфологических изменений сердца. Проведение МРТ сердца у пациентов с акромегалий расширяет наше представление о развитии НРПС при данном заболевании. В настоящее время отсутствуют специфические лабораторные маркеры диффузного миокардиального фиброза, а его роль в возникновении нарушений ритма и проводимости сердца требует дальнейшего изучения.

Antitumor and immunotropic effects of κ-, λ-carrageenan from red marine algae Chondrus armatus and their low-molecular weight (LMW) degradation products were explored. Effects on human esophageal cancer cell lines KYSE30 and FLO1 viability and ability to induce production of pro- and anti-inflammatory cytokines by human monocytes was assessed. All polysaccharides demonstrated antimetabolic and cytostatic activity towards cancer lines, with high-molecular weight carrageenans possessing higher antimetabolic and lower cytostatic activity than their LMW degradation products. All carrageenans induced monocytes to produce pro-inflammatory cytokines IL1β, IL6, IL18, and TNFα. However, secretion of anti-inflammatory cytokine IL10 was induced only by LMW λ-carrageenan, which exhibited the highest cytokine production inducing efficacy overall. We demonstrate that LMW carrageenan degradation products not only retain biological activity of their precursors, but also increase their efficacy in type-dependent manner, allowing for their future development for pharmacological practice.

Circulating autoantibodies against tumor-associated autoantigens (TAA) may serve as valuable biomarkers for a wide range of diagnostic purposes. Modern immunology offers a large variety of methods for in-depth comparative analysis of the repertoires of circulating antibodies' antigenic specificities in health and disease. Nevertheless, this research field so far has met somewhat limited clinical success, while numerous data on the repertoires of circulating autoantibodies' specificities in cancer patients are poorly integrated into the contemporary picture of the immunological and molecular landscapes of human tumors. This review is an attempt to identify and systematize the key and essentially universal conceptual and methodological limitations of analyses of the repertoires of circulating antibodies' antigenic specificities in cancer (expression bias, redundancy of TAA repertoires, identification of natural IgG, the absence of the pathogenetically relevant context in the experimental systems used to detect TAA), as well as to discuss potential and already known methodological improvements that may significantly increase the detectability of the pathogenetically relevant and diagnostically significant bona fide TAA.

Detection of thymidine analogues after their incorporation into replicating DNA represents a powerful tool for the study of cellular DNA synthesis, progression through the cell cycle, cell proliferation kinetics, chronology of cell division, and cell fate determination. Recent advances in the concurrent detection of multiple such analogues offer new avenues for the investigation of unknown features of these vital cellular processes. Combined with quantitative analysis, temporal discrimination of multiple labels enables elucidation of various aspects of stem cell life cycle in situ, such as division modes, differentiation, maintenance, and elimination. Data obtained from such experiments are critically important for creating descriptive models of tissue histogenesis and renewal in embryonic development and adult life. Despite the wide use of thymidine analogues in stem cell research, there are a number of caveats to consider for obtaining valid and reliable labeling results when marking replicating DNA with nucleotide analogues. Therefore, in this review, we describe critical points regarding dosage, delivery, and detection of nucleotide analogues in the context of single and multiple labeling, outline labeling schemes based on pulse-chase, cumulative and multilabel marking of replicating DNA for revealing stem cell proliferative behaviors, and determining cell cycle parameters, and discuss preconditions and pitfalls in conducting such experiments. The information presented in our review is important for rational design of experiments on tracking dividing stem cells by marking replicating DNA with thymidine analogues.

Insoluble protein aggregates with fibrillar morphology called amyloids and β-barrel proteins both share a β-sheet-rich structure. Correctly folded β-barrel proteins can not only function in monomeric (dimeric) form, but also tend to interact with one another—followed, in several cases, by formation of higher order oligomers or even aggregates. In recent years, findings proving that β-barrel proteins can adopt cross-β amyloid folds have emerged. Different β-barrel proteins were shown to form amyloid fibrils in vitro. The formation of functional amyloids in vivo by β-barrel proteins for which the amyloid state is native was also discovered. In particular, several prokaryotic and eukaryotic proteins with β-barrel domains were demonstrated to form amyloids in vivo, where they participate in interspecies interactions and nutrient storage, respectively. According to recent observations, despite the variety of primary structures of amyloid-forming proteins, most of them can adopt a conformational state with the β-barrel topology. This state can be intermediate on the pathway of fibrillogenesis (“on-pathway state”), or can be formed as a result of an alternative assembly of partially unfolded monomers (“off-pathway state”). The β-barrel oligomers formed by amyloid proteins possess toxicity, and are likely to be involved in the development of amyloidoses, thus representing promising targets for potential therapy of these incurable diseases. Considering rapidly growing discoveries of the amyloid-forming β-barrels, we may suggest that their real number and diversity of functions are significantly higher than identified to date, and represent only “the tip of the iceberg”. Here, we summarize the data on the amyloid-forming β-barrel proteins, their physicochemical properties, and their biological functions, and discuss probable means and consequences of the amyloidogenesis of these proteins, along with structural relationships between these two widespread types of β-folds.

Surgery of the ureterovesical anastomoses lesions in case of weakly-dilated ureters is well developed by the Cohen and Lich-Grgoire techniques. However, there are no generally accepted approaches for heavily-dilated ureters. The experience of creating ureterocystoneoanastomosis according to the Bradi technique (1975) for the first time in the literature is presented.The authors modification of the Bradi technique was used in 12 patients of both sexes with a dilated ureters diameter of more than 10 mm. Two cases of refluxogenic and 10 cases of obstructive megaureter were operated on 2010-2019. The follow-up period was 1-10 years. All patients underwent resection of the ureter in width.No intraoperative complications were noted. One case of acute pyelonephritis with acute urinary retention was noted as postoperative complications. All patients showed a decrease in dilatation of the pyelocaliceal system and ureter according to multispiral computed tomography after 6 months of follow-up. Two patients had vesicoureteral reflux of the first degree. Glomerular filtration rates decreased in 41.7% of cases after 12 months of follow-up. No recurrence of urinary tract infection and stenosis of the anastomosis was detected for 1-10 years monitoring.The Bradi technique along with the Hodgsons ureter reconstruction provides superior results for heavily-dilated ureters in adults.

Adrenocortical cancer (ACC) is a rare endocrine malignancy of the adrenal cortex, which has an unfavorable prognosis and extremely aggressive clinical behavior in most cases. Nevertheless, cases of a more favorable disease course with late metastasis and slow progression have been described. In 2017, the International Agency for Research on Cancer (IARC) and the World Health Organization (WHO) in the 4To provide a detailed description of the morphological variants of ACC with an emphasis on their histological characteristics and the expression of immunohistochemical markers.A total of 75 cases of ACC were analyzed in the adult population diagnosed as having the morphological variants in accordance with the International Histological Classification of Adrenal Tumors (WHO, 2017). Monoclonal antibodies to SF1, Inhibin A, Melan A, Ki-67, p53, and antimitochondrial antibodies were used for immunohistochemical diagnosis.The classic, oncocytic, and myxoid subtypes of ACC were found in 51 (68%), 15 (20%), and 9 (12%) cases, respectively. The functional activity of the tumors was observed in 43% (Despite the determination of a minimal immunohistochemical panel to confirm the diagnosis of ACC, it is important to remember that each histological variant may be characterized by the different expression of immunohistochemical markers. The identification of morphological variants of ACC and the use of specific, sensitive, and prognostically significant immunohistochemical markers will allow clinicians and pathologists to more accurately judge the biological properties of this tumor and the clinical course of the disease.Адренокортикальный рак (АКР) представляет собой редкую злокачественную эндокринную опухоль коры надпочечников, в большинстве случаев имеющую неблагоприятный прогноз и крайне агрессивное клиническое поведение. Тем не менее описаны случаи более благоприятного течения заболевания с поздним метастазированием и медленным прогрессированием. В 2017 г. Международным агентством по изучению рака (International Agency for Research on Cancer, IARC) и Всемирной организацией здравоохранения (ВОЗ) в 4-м издании Классификации опухолей эндокринных органов (WHO Classification of Tumors Pathology and Genetics) выделены гистологические варианты АКР: классический, онкоцитарный, миксоидный и саркоматоидный, свидетельствующие о морфологической неоднородности такой опухоли.Подробное описание морфологических вариантов АКР с акцентом на их гистологических характеристиках и экспрессии иммуногистохимических маркеров.В настоящем исследовании выполнен анализ 75 случаев АКР во взрослой популяции с диагностикой морфологических вариантов в соответствии с Международной гистологической классификацией опухолей надпочечника (ВОЗ, 2017 г.). Для иммуногистохимической диагностики использованы моноклональные антитела к SF1, Inhibin A, Melan A, Ki-67, р53, антимитохондриальные антитела.Классический подтип АКР выявлен у 51 (68%) человека, онкоцитарный — у 15 (20%), миксоидный — у 9 (12%). Функциональная активность опухолей наблюдалась в 43% (18 случаев) при классическом варианте АКР, при этом наиболее часто клиническая картина проявлялась симптомами гиперкортицизма (38%) и вирилизации (5%). Достоверных различий в гормональной активности между различными морфологическими вариантами не обнаружено. Определена характеристика вышеописанных гистологических вариантов опухоли с описанием паттернов роста, которые позволяют улучшить диагностику АКР. Подтверждение диагноза АКР возможно только при проведении иммуногистохимического исследования, необходимая минимальная панель маркеров должна включать: SF1, Melan A, Inhibin A. Индекс пролиферативной активности Ki-67 показал достоверные различия (Несмотря на определение минимальной иммуногистохимической панели, позволяющей подтвердить диагноз АКР, важно помнить, что каждый гистологический вариант может характеризоваться различной выраженностью экспрессии иммуногистохимических маркеров. Выделение морфологических вариантов АКР и использование специфичных, чувствительных и прогностически значимых иммуногистохимических маркеров позволят клиницистам и патоморфологам более точно судить о биологических свойствах этой опухоли и клиническом течении заболевания.

A course of treatment with humic acids extracted with sodium pyrophosphate from high-moor pine-peat moss-cotton grass peat improves humoral immune response of C57BL/6 mice, stimulates the production of TNFα, IL-1β, and IL-12 by the animal peritoneal macrophages and the production of IFNγ and TNFα by donor peripheral blood mononuclears, causing no changes in the production of IL-10 in vitro.

The Q61R mutation of the NRAS gene is one of the most frequent driver mutations of thyroid cancer. Tumors with this mutation are characterized by invasion into blood vessels and formation of distant metastases. To study the role of this mutation in the growth of thyroid cancer, we developed a model system on the basis of thyroid epithelial cell line Nthy-ori 3-1 transduced by a lentiviral vector containing the NRAS gene with the Q61R mutation. It was found that the expression of NRAS(Q61R) in thyroid epithelial cells has a profound influence on groups of genes involved in the formation of intercellular contacts, as well as in processes of epithelial-mesenchymal transition and cell invasion. The alteration in the expression of these genes affects the phenotype of the model cells, which acquire traits of mesenchymal cells and demonstrate increased ability for survival and growth without attachment to the substrate. The key regulators of these processes are transcription factors belonging to families SNAIL, ZEB, and TWIST, and in different types of tumors the contribution of each individual factor can vary greatly. In our model system, phenotype change correlates with an increase in the expression of SNAIL2 and TWIST2 factors, which indicates their possible role in regulating invasive growth of thyroid cancer with the mutation of NRAS(Q61R).

PURPOSE Glofitamab is a T-cell–engaging bispecific antibody possessing a novel 2:1 structure with bivalency for CD20 on B cells and monovalency for CD3 on T cells. This phase I study evaluated glofitamab in relapsed or refractory (R/R) B-cell non-Hodgkin lymphoma (B-NHL). Data for single-agent glofitamab, with obinutuzumab pretreatment ( Gpt) to reduce toxicity, are presented. METHODS Seven days before the first dose of glofitamab (0.005-30 mg), all patients received 1,000 mg Gpt. Dose-escalation steps were determined using a Bayesian continuous reassessment method with overdose control. Primary end points were safety, pharmacokinetics, and the maximum tolerated dose of glofitamab. RESULTS Following initial single-patient cohorts, 171 patients were treated within conventional multipatient cohorts and received at least one dose of glofitamab. This trial included heavily pretreated patients with R/R B-NHL; most were refractory to prior therapy (155; 90.6%) and had received a median of three prior therapies. One hundred and twenty-seven patients (74.3%) had diffuse large B-cell lymphoma, transformed follicular lymphoma, or other aggressive histology, and the remainder had indolent lymphoma subtypes. Five (2.9%) patients withdrew from treatment because of adverse events. Cytokine release syndrome occurred in 86 of 171 (50.3%) patients (grade 3 or 4: 3.5%); two (1.2%) patients experienced grade 3, transient immune effector cell–associated neurotoxicity syndrome-like symptoms. The overall response rate was 53.8% (complete response [CR], 36.8%) among all doses and 65.7% (CR, 57.1%) in those dosed at the recommended phase II dose. Of 63 patients with CR, 53 (84.1%) have ongoing CR with a maximum of 27.4 months observation. CONCLUSION In patients with predominantly refractory, aggressive B-NHL, glofitamab showed favorable activity with frequent and durable CRs and a predictable and manageable safety profile.

Distal ureter strictures are complications of surgical interventions for obstetric, gynecological, urological, vascular and oncological diseases. Reconstructive plastic correction of these strictures is a complex urological procedure. The choice of a technique is associated with the localization and extent of the stricture. Boari flap ureteroneocystostomy is one of the main operations to choose from. Its implementation in open and laparoscopic versions in recent history required special analysis.to analyze the experience of ureteral reimplantation according to the Boari technique with a detailed description of its open and laparoscopic approaches.In 2010-2019, 30 patients underwent surgery using the Boari technique. 17 patients underwent open surgery (group 1) and 13 patients underwent surgery using laparoscopic techniques (group 2). A comparative analysis of the two groups was carried out considering clinical and intraoperative data. Complications were studied, as well as immediate and long-term results with a follow-up period of 6-120 months.Patients of the two groups showed similar clinical characteristics. Patients had an equal extent of lesion in the groups (p0.05). However, the lesions were located more proximally from the bladder in group 1, which required the use of longer flaps (p=0.024). Blood loss volume was identical, and the duration of laparoscopic operations was significantly shorter (p=0.019). Postoperative complications occurred in 26.7% of cases in the next 3 months, which required a temporary percutaneous puncture nephrostomy (IIIa degree according to Clavien-Dindo) in 2 cases and conservative therapy (II degree according to Clavien-Dindo) in 6 cases. Clinical vesicoureteral reflux was determined in only one case during a one-year follow-up period.Open and laparoscopic Boari techniques have an equally high ureteral recovery efficiency without the need for a re-operation in all cases. Refluxing flap ureteral anastomosis is extremely rarely accompanied by a clinic of vesicoureteral reflux. The ureter can be restored using a laparoscopic Boari technique in all cases.

Genetically-encoded fluorescent sensors have been actively developed over the last few decades and used in live imaging and drug screening. Real-time monitoring of drug action in a specific cellular compartment, organ, or tissue type; the ability to screen at the single-cell resolution; and the elimination of false-positive results caused by low drug bioavailability that is not detected by in vitro testing methods are a few of the obvious benefits of using genetically-encoded fluorescent sensors in drug screening. In combination with high-throughput screening (HTS), some genetically-encoded fluorescent sensors may provide high reproducibility and robustness to assays. We provide a brief overview of successful, perspective, and hopeful attempts at using genetically encoded fluorescent sensors in HTS of modulators of ion channels, Ca2+ homeostasis, GPCR activity, and for screening cytotoxic, anticancer, and anti-parasitic compounds. We discuss the advantages of sensors in whole organism drug screening models and the perspectives of the combination of human disease modeling by CRISPR techniques with genetically encoded fluorescent sensors for drug screening.

Background Recent studies reported abnormal alpha-synuclein deposition in biopsy-accessible sites of the peripheral nervous system in Parkinson's disease (PD). This has considerable implications for clinical diagnosis. Moreover, if deposition occurs early, it may enable tissue diagnosis of prodromal PD. Objective The aim of this study was to develop and test an automated bright-field immunohistochemical assay of cutaneous pathological alpha-synuclein deposition in patients with idiopathic rapid eye movement sleep behavior disorder, PD, and atypical parkinsonism and in control subjects. Methods For assay development, postmortem skin biopsies were taken from 28 patients with autopsy-confirmed Lewy body disease and 23 control subjects. Biopsies were stained for pathological alpha-synuclein in automated stainers using a novel dual-immunohistochemical assay for serine 129-phosphorylated alpha-synuclein and pan-neuronal marker protein gene product 9.5. After validation, single 3-mm punch skin biopsies were taken from the cervical 8 paravertebral area from 79 subjects (28 idiopathic rapid eye movement sleep behavior disorder, 20 PD, 10 atypical parkinsonism, and 21 control subjects). Raters blinded to clinical diagnosis assessed the biopsies. Results The immunohistochemistry assay differentiated alpha-synuclein pathology from nonpathological-appearing alpha-synuclein using combined phosphatase and protease treatments. Among autopsy samples, 26 of 28 Lewy body samples and none of the 23 controls were positive. Among living subjects, punch biopsies were positive in 23 (82%) subjects with idiopathic rapid eye movement sleep behavior disorder, 14 (70%) subjects with PD, 2 (20%) subjects with atypical parkinsonism, and none (0%) of the control subjects. After a 3-year follow-up, eight idiopathic rapid eye movement sleep behavior disorder subjects phenoconverted to defined neurodegenerative syndromes, in accordance with baseline biopsy results. Conclusion Even with a single 3-mm punch biopsy, there is considerable promise for using pathological alpha-synuclein deposition in skin to diagnose both clinical and prodromal PD. © 2020 International Parkinson and Movement Disorder Society.

Summary: Store-operated calcium entry (SOCE) through STIM-gated ORAI channels governs vital cellular functions. In this context, SOCE controls cellular redox signaling and is itself regulated by redox modifications. However, the molecular mechanisms underlying this calcium-redox interplay and the functional outcomes are not fully understood. Here, we examine the role of STIM2 in SOCE redox regulation. Redox proteomics identify cysteine 313 as the main redox sensor of STIM2 in vitro and in vivo. Oxidative stress suppresses SOCE and calcium currents in cells overexpressing STIM2 and ORAI1, an effect that is abolished by mutation of cysteine 313. FLIM and FRET microscopy, together with MD simulations, indicate that oxidative modifications of cysteine 313 alter STIM2 activation dynamics and thereby hinder STIM2-mediated gating of ORAI1. In summary, this study establishes STIM2-controlled redox regulation of SOCE as a mechanism that affects several calcium-regulated physiological processes, as well as stress-induced pathologies.

Redox reactions are of high fundamental and practical interest since they are involved in both normal physiology and the pathogenesis of various diseases. However, this area of research has always been a relatively problematic field in the context of analytical approaches, mostly because of the unstable nature of the compounds that are measured. Genetically encoded sensors allow for the registration of highly reactive molecules in real-time mode and, therefore, they began a new era in redox biology. Their strongest points manifest most brightly in in vivo experiments and pave the way for the non-invasive investigation of biochemical pathways that proceed in organisms from different systematic groups. In the first part of the review, we briefly describe the redox sensors that were used in vivo as well as summarize the model systems to which they were applied. Next, we thoroughly discuss the biological results obtained in these studies in regard to animals, plants, as well as unicellular eukaryotes and prokaryotes. We hope that this work reflects the amazing power of this technology and can serve as a useful guide for biologists and chemists who work in the field of redox processes.

Analysis of development of «robotic surgery» on the basis of historical prerequisites, review of the possibilities of the new Senhance robotic surgical system and determination of further perspective.A prospective group of 51 patients with surgical pathology who underwent surgical interventions using the new Senhance robotic system was analyzed. The study included 51 patients. Robot-assisted cholecystectomy was performed in 20 patients, 31 patients underwent robotic transabdominal preperitoneal plastic surgery.A short-term analysis of intraoperative and early postoperative data showed the safety of using the Senhance robotic complex for the patient. The ergonomics of this system contributes to significantly less fatigue of the surgeon during the operation, increasing its efficiency and safety.The emergence of competitive robotic systems contributes to the dynamic development of robotic surgery, which in turn will lead to a reduction in the cost of these technologies in the future. Innovative solutions used in modern robotic complexes are based on the principles of machine learning, which in due time will make the «robot surgeon» not just a tool, but a full-fledged member of the operating team.На основе исторических предпосылок провести анализ развития роботизированной хирургии, изучить возможности новой роботической хирургической системы Senhance и определить дальнейшие перспективы ее применения.Проанализированы данные проспективной группы, включающей 51 пациента с хирургической патологией, которым были проведены оперативные вмешательства с применением новой роботической системы Senhance. Робот-ассистированная холецистэктомия выполнена у 20 пациентов, роботическая трансабдоминальная преперитонеальная пластика паховой грыжи — у 31 пациента.Анализ интраоперационных и ранних послеоперационных данных показал безопасность применения для пациентов роботического комплекса Senhance. Эргономичность системы способствует существенно меньшему утомлению хирурга в ходе операции, увеличивает ее эффективность.Появление конкурентоспособных роботических систем способствует динамичному развитию роботизированный хирургии и в перспективе приведет к снижению стоимости использования этой технологии. Инновационные решения, применяемые в современных роботических комплексах, основываются на принципах машинного обучения, что в дальнейшем сделает «робота-хирурга» не просто инструментом, а полноценном членом операционной бригады.

Amyloids are fibrillar protein aggregates with an ordered spatial structure called “cross-β”. While some amyloids are associated with development of approximately 50 incurable diseases of humans and animals, the others perform various crucial physiological functions. The greatest diversity of amyloids functions is identified within prokaryotic species where they, being the components of the biofilm matrix, function as adhesins, regulate the activity of toxins and virulence factors, and compose extracellular protein layers. Amyloid state is widely used by different pathogenic bacterial species in their interactions with eukaryotic organisms. These amyloids, being functional for bacteria that produce them, are associated with various bacterial infections in humans and animals. Thus, the repertoire of the disease-associated amyloids includes not only dozens of pathological amyloids of mammalian origin but also numerous microbial amyloids. Although the ability of symbiotic microorganisms to produce amyloids has recently been demonstrated, functional roles of prokaryotic amyloids in host–symbiont interactions as well as in the interspecies interactions within the prokaryotic communities remain poorly studied. Here, we summarize the current findings in the field of prokaryotic amyloids, classify different interspecies interactions where these amyloids are involved, and hypothesize about their real occurrence in nature as well as their roles in pathogenesis and symbiosis.

With the growing interest of the medical industry in biocompatible nanoparticles (NPs), the current synthetic methods should be adapted to appropriate demands (toxicity, scalability, etc.). Most applications require colloidal systems to be stable not only in water but also in vivo, which represents a major challenge. In this study, biocompatible Ta2O5 NPs were synthesized by a solvothermal method avoiding toxic reagents, and surfactant-free stable hydrosols were obtained and used for computed tomography (CT) imaging. The small hydrodynamic size (2 nm) and colloidal stability of primary NPs were studied by dynamic light scattering (DLS). The particles were characterized by X-ray diffraction, transmission electron microscopy, energy-dispersive X-ray spectroscopy, and Brunauer-Emmett-Teller analysis to confirm their structure and purity. To develop a stable hydrosol preparation protocol, the influence of pH and ultrasonication duration on the stability of Ta2O5 sols was analyzed by DLS and microelectrophoresis. To enhance the understanding of NP behavior in vivo, sol stability in conditions close to physiological (NaCl solutions) was studied in a pH range of 3-9. Hydrosols prepared by the proposed protocol were stable for at least 6 months and exhibited negligible cytotoxicity. Ta2O5 NPs also showed high CT contrast both in theoretical calculations and in vivo (rat gastrointestinal tract).

The root cause of non-inherited Alzheimer’s disease (AD) remains unknown despite hundreds of research studies performed to attempt to solve this problem. Since proper prophylaxis remains the best strategy, many scientists have studied the risk factors that may affect AD development. There is robust evidence supporting the hypothesis that cardiovascular diseases (CVD) may contribute to AD progression, as the diseases often coexist. Therefore, a lack of well-defined diagnostic criteria makes studying the relationship between AD and CVD complicated. Additionally, inflammation accompanies the pathogenesis of AD and CVD, and is not only a consequence but also implicated as a significant contributor to the course of the diseases. Of note, АроЕε4 is found to be one of the major risk factors affecting both the cardiovascular and nervous systems. According to genome wide association and epidemiological studies, numerous common risk factors have been associated with the development of AD-related pathology. Furthermore, the risk of developing AD and CVDs appears to be increased by a wide range of conditions and lifestyle factors: hypertension, dyslipidemia, hypercholesterolemia, hyperhomocysteinemia, gut/oral microbiota, physical activity, and diet. This review summarizes the literature and provides possible mechanistic links between CVDs and AD.

Hydrogen peroxide (H2O2) modulates critical phosphorylation pathways in vascular endothelial cells, many of which affect endothelial nitric oxide synthase (eNOS) signal transduction. Both intracellular and extracellular sources of H2O2 have been implicated in eNOS regulation, yet the specific endothelial pathways remain incompletely understood. Here we exploited chemogenetic approaches and live-cell imaging methods to both generate and detect H2O2 in different subcellular compartments (cytosol, nucleus, and caveolae) of cultured EA.hy926 human endothelial cells. We developed novel recombinant constructs encoding differentially-targeted yeast d-amino acid oxidase (DAAO), which generates H2O2 only when its d-amino acid substrate is provided. DAAO was expressed as a fusion protein with the new H2O2 biosensor HyPer7.2, which allowed us to quantitate intracellular H2O2 levels by ratiometric imaging in living endothelial cells following the activation of DAAO by d-alanine. The addition of extracellular H2O2 to the HyPer-DAAO-transfected cells led to increases in H2O2 throughout different regions of the cell, as measured using the differentially-targeted HyPer biosensor for H2O2. The sensor response to extracellular H2O2 was more rapid than that quantitated following the addition of d-alanine to transfected cells to activate differentially-targeted DAAO. The maximal intracellular levels of H2O2 observed in response to the addition of extracellular H2O2 vs. intracellular (DAAO-generated) H2O2 were quantitatively similar. Despite these similarities in the measured levels of intracellular H2O2, we observed a remarkable quantitative difference in the activation of endothelial phosphorylation pathways between chemogenetically-generated intracellular H2O2 and the phosphorylation responses elicited by the addition of extracellular H2O2 to the cells. Addition of extracellular H2O2 had only a nominal effect on phosphorylation of eNOS, kinase Akt or AMP-activated protein kinase (AMPK). By contrast, intracellular H2O2 generation by DAAO caused striking increases in the phosphorylation of these same key signaling proteins. We also found that the AMPK inhibitor Compound C completely blocked nuclear H2O2-promoted eNOS phosphorylation. However, Compound C had no effect on eNOS phosphorylation following H2O2 generation from cytosol- or caveolae-targeted DAAO. We conclude that H2O2 generated in the cell nucleus activates AMPK, leading to eNOS phosphorylation; in contrast, AMPK activation by cytosol- or caveolae-derived H2O2 does not promote eNOS phosphorylation via AMPK. These findings indicate that H2O2 generated in different subcellular compartments differentially modulates endothelial cell phosphorylation pathways, and suggest that dynamic subcellular localization of oxidants may modulate signaling responses in endothelial cells.

Natural oxygen gradients occur in tissues of biological organisms and also in the context of three-dimensional (3D) in vitro cultivation. Oxygen diffusion limitation and metabolic oxygen consumption by embedded cells produce areas of hypoxia in the tissue/matrix. However, reliable systems to detect oxygen gradients and cellular response to hypoxia in 3D cell culture systems are still missing. In this study, we developed a system for visualization of oxygen gradients in 3D using human adipose tissue–derived mesenchymal stem cells (hAD-MSCs) modified to stably express a fluorescent genetically engineered hypoxia sensor HRE-dUnaG. Modified cells retained their stem cell characteristics in terms of proliferation and differentiation capacity. The hypoxia-reporter cells were evaluated by fluorescence microscopy and flow cytometry under variable oxygen levels (2.5%, 5%, and 7.5% O2). We demonstrated that reporter hAD-MSCs output is sensitive to different oxygen levels and displays fast decay kinetics after reoxygenation. Additionally, the reporter cells were encapsulated in bulk hydrogels with a variable cell number, to investigate the sensor response in model 3D cell culture applications. The use of hypoxia-reporting cells based on MSCs represents a valuable tool for approaching the genuine in vivo cellular microenvironment and will allow a better understanding of the regenerative potential of AD-MSCs. © 2020 The Authors. Biotechnology and Bioengineering published by Wiley Periodicals LLC

PD-L1/PD-1 blocking antibodies have demonstrated therapeutic efficacy across a range of human cancers. Extending this benefit to a greater number of patients, however, will require a better understanding of how these therapies instigate anticancer immunity. Although the PD-L1/PD-1 axis is typically associated with T cell function, we demonstrate here that dendritic cells (DCs) are an important target of PD-L1 blocking antibody. PD-L1 binds two receptors, PD-1 and B7.1 (CD80). PD-L1 is expressed much more abundantly than B7.1 on peripheral and tumor-associated DCs in patients with cancer. Blocking PD-L1 on DCs relieves B7.1 sequestration in cis by PD-L1, which allows the B7.1/CD28 interaction to enhance T cell priming. In line with this, in patients with renal cell carcinoma or non-small cell lung cancer treated with atezolizumab (PD-L1 blockade), a DC gene signature is strongly associated with improved overall survival. These data suggest that PD-L1 blockade reinvigorates DC function to generate potent anticancer T cell immunity.

Hydrogen peroxide (H(2)O(2)) is a key redox intermediate generated within cells. Existing probes for H(2)O(2) have not solved the problem of detection of the ultra-low concentrations of the oxidant: these reporters are either not sensitive enough, or pH-dependent, or insufficiently bright, or not functional in mammalian cells, or have poor dynamic range. Here we present HyPer7, the first bright, pH-stable, ultrafast and ultrasensitive ratiometric H(2)O(2) probe. HyPer7 is fully functional in mammalian cells and in other higher eukaryotes. The probe consists of a circularly permuted GFP integrated into the ultrasensitive OxyR domain from Neisseria meningitidis. Using HyPer7, we were able to uncover the details of H(2)O(2) diffusion from the mitochondrial matrix, to find a functional output of H(2)O(2) gradients in polarized cells, and to prove the existence of H(2)O(2) gradients in wounded tissue in vivo. Overall, HyPer7 is a probe of choice for real-time H(2)O(2) imaging in various biological contexts.

Antigen recognition by T-cells is guided by the T-cell receptor (TCR) heterodimer formed by α and β chains. A huge diversity of TCR sequences should be maintained by the immune system in order to be able to mount an effective response towards foreign pathogens, so, due to cooperative binding of α and β chains to the pathogen, any constraints on chain pairing can have a profound effect on immune repertoire structure, diversity and antigen specificity. By integrating available structural data and paired chain sequencing results we were able to show that there are almost no constraints on pairing in TCRαβ complexes, allowing naive T-cell repertoire to reach the highest possible diversity. Additional analysis reveals that the specific choice of contacting amino acids can still have a profound effect on complex conformation. Moreover, antigen-driven selection can distort the uniform landscape of chain pairing, while small, yet significant, differences in the pairing can be attributed to various specialized T-cell subsets such as MAIT and iNKT T-cells, as well as other TCR sets specific to certain antigens., Author summary In the present paper we study chain pairing preferences in the T-cell receptor (TCR) heterodimer complex. The TCR molecule is formed by α and β chains and binding of both of these chains to an antigen presented by the major histocompatibility complex (MHC) molecule is required in order to trigger an immune response against foreign pathogens and neoantigens. We show that chain pairing in the TCR complex is nearly random ensuring a highly diverse set of TCRs required for recognition of a vast set of antigens. Our results also show that chain pairing preferences can nevertheless influence TCR complex geometry and biases in TCR chain pairing can be used to identify antigen-driven selection or selection towards specialized subsets of T-cells such as mucosal-associated and natural killer invariant T-cells.

Increasing numbers of patients are sharing their health-related experiences online in forums, or on social media websites, such as Twitter and Facebook. This largely untapped source of data about patients’ experience of living with disease and its treatment may be useful in deriving drug safety information such as the occurrence, nature and impact of side effects. Text mining techniques can transform free text into structured data amenable for analysis by automatically recognising mentions of various health conditions and their relationship to a particular medication. These techniques have been used to identify the occurrence of commonly discussed drug adverse events (AEs) from posts on Facebook and Twitter.1 2 They have also been used to identify discussions about benefits of drugs and how these benefits compared with the AEs, other treatment options, costs and complaints about the product.1 A recently published analysis of Twitter posts mentioning prednisolone or prednisone found insomnia and weight gain to be the most frequently discussed side effects.3 However, with the 140 (or more recently 280) character limit per tweet, any side effect information is limited to what can be included and discussed within this space. HealthUnlocked (HU), Europe’s largest social media network for health that supports patients and healthcare providers, hosts over 700 communities (including the UK’s …

Postoperative stress urinary incontinence has a significant effect on reducing the quality of life of patients undergoing radical prostatectomy. This review discusses the existing methods for correcting this condition, described in the world literature and which to one degree or another improve the quality of life of patients in this category. Analysis of present data shows that currently there are 4 main basic techniques during radical prostatectomy, which allow to statistically significantly improve of functional results. In particular, usage of this methodic allows reducing the frequency and degree of postoperative incontinence: 1) preservation of neurovascular bundles, 2) anterior and/or posterior reconstruction of the fascial spaces of the pelvis, 3) preservation of the maximum length of the membranous urethra, 4) reconstruction of the bladder neck. Preservation of the neurovascular bundles, anterior and/or posterior reconstruction of the pelvic fascial spaces have already proven themselves as effective techniques currently used by many surgeons. In contrast, the clinical utility of reserve the maximum possible length of the membranous urethra and reconstructive surgery of the bladder neck are relatively new techniques. Their clinical usefulness, as well as safety, continues to be studied.

Summary: Zyxin is a cytoskeletal LIM-domain protein that regulates actin cytoskeleton assembly and gene expression. In the present work, we find that zyxin downregulation in Xenopus laevis embryos reduces the expression of numerous genes that regulate cell differentiation, but it enhances that of several genes responsible for embryonic stem cell status, specifically klf4, pou5f3.1, pou5f3.2, pou5f3.3, and vent2.1/2. For pou5f3 family genes (mammalian POU5F1/OCT4 homologs), we show that this effect is the result of mRNA stabilization due to complex formation with the Y-box protein Ybx1. When bound to Ybx1, zyxin interferes with the formation of these complexes, thereby stimulating pou5f3 mRNA degradation. In addition, in zebrafish embryos and human HEK293 cells, zyxin downregulation increases mRNA levels of the pluripotency genes KLF4, NANOG, and POU5F1/OCT4. Our findings indicate that zyxin may play a role as a switch among morphogenetic cell movement, differentiation, and embryonic stem cell status.

Effects of testosterone (T) on the cardiovascular system of men remain controversial. The impact of T-replacement therapy (TRT) in men with functional hypogonadism and type 2 diabetes mellitus (T2DM) has to be elucidated. This study included 80 men (mean age 51.5 ± 6.3 years) with newly diagnosed T2DM (according to ADA criteria) and functional hypogonadism (according to EAU criteria). Randomization: Group1 (

Described herein is a clinical case report regarding treatment of a 70-year-old male patient presenting with a late complication following endoprosthetic repair for a Stanford type B dissecting thoracic aortic aneurysm. The man was admitted to our hospital for persistent type IIb endoleak and an increased diameter of the aorta in its thoracic and thoracoabdominal portions. Two years previously, he had endured endoprosthetic repair of the thoracic aorta. The findings of computed tomography revealed negative dynamics manifesting as an increase in the diameter of the false channel of the arch and descending thoracic aorta with persistent type IIb endoleak. He was subjected to elimination of abdominal aortic dissection and type IIb endoleak with partial prosthetic repair of the descending thoracic portion of the aorta by means of prosthetic repair of the lower thoracic portion of the aorta between the stent graft and linear vascular Dacron prosthesis. The postoperative period was complicated by transient acute renal failure and paraparesis of the lower limbs. The patient was discharged on POD 14, with no endoleaks revealed on control computed tomography 3 months thereafter.Представлен случай лечения пациента 70 лет с поздним осложнением после эндопротезирования грудной аорты при расслаивающейся аневризме B типа по Стэнфорду. Больной поступил в стационар по поводу сохранения эндолика Iб типа и увеличения диаметра аорты в ее грудном и торакоабдоминальном отделах. Пациент два года назад перенес эндопротезирование грудной аорты. При компьютерной томографии выявлена отрицательная динамика в виде увеличения диаметра ложного канала дуги и нисходящей грудной аорты с сохранением эндолика Ib типа. Выполнена ликвидация расслоения брюшного отдела аорты и эндолика Iб типа с частичным протезированием нисходящего грудного отдела аорты путем протезирования нижнегрудного отдела аорты между стент-графтом и линейным сосудистым дакроновым протезом. Послеоперационный период осложнился транзиторной острой почечной недостаточностью и парапарезом нижних конечностей. Выписан на 14 сутки после операции. При контрольной компьютерной томографии через 3 месяца эндоликов не выявлено.

Neuronal gap junctional protein connexin 36 (Cx36) contributes to neuronal death following a range of acute brain insults such as ischemia, traumatic brain injury and epilepsy. Whether Cx36 contributes to neuronal death and pathological outcomes in chronic neurodegenerative diseases, such as amyotrophic lateral sclerosis (ALS), is not known. We show here that the expression of Cx36 is significantly decreased in lumbar segments of the spinal cord of both human ALS subjects and SOD1G93A mice as compared to healthy human and wild-type mouse controls, respectively. In purified neuronal cultures prepared from the spinal cord of wild-type mice, knockdown of Cx36 reduces neuronal death caused by overexpression of the mutant human SOD1-G93A protein. Taken together, these data suggest a possible contribution of Cx36 to ALS pathogenesis. A perspective for the use of blockers of Cx36 gap junction channels for ALS therapy is discussed.

Multimodal nonlinear microscopy combining third-harmonic generation (THG) with two- and three-photon-excited fluorescence (2PEF and 3PEF) is shown to provide a powerful resource for high-fidelity imaging of nucleoli and nucleolar proteins. We demonstrate that, with a suitably tailored genetically encoded fluorescent stain, the 2PEF/3PEF readout from specific nucleolar proteins can be reliably detected against the extranucleolar 2PEF/3PEF signal, enabling high-contrast imaging of the key nucleolar ribosome biogenesis components, such as fibrillarin. THG is shown to provide a versatile readout for unstained nucleolus imaging in a vast class of biological systems as different as neurons in brain slices and cultured HeLa cells.

Thermogenetics is a promising innovative neurostimulation technique, which enables robust activation of neurons using thermosensitive transient receptor potential (TRP) cation channels. Broader application of this approach in neuroscience is, however, hindered by a limited variety of suitable ion channels, and by low spatial and temporal resolution of neuronal activation when TRP channels are activated by ambient temperature variations or chemical agonists. Here, we demonstrate rapid, robust and reproducible repeated activation of snake TRPA1 channels heterologously expressed in non-neuronal cells, mouse neurons and zebrafish neurons in vivo by infrared (IR) laser radiation. A fibre-optic probe that integrates a nitrogen−vacancy (NV) diamond quantum sensor with optical and microwave waveguide delivery enables thermometry with single-cell resolution, allowing neurons to be activated by exceptionally mild heating, thus preventing the damaging effects of excessive heat. The neuronal responses to the activation by IR laser radiation are fully characterized using Ca2+ imaging and electrophysiology, providing, for the first time, a complete framework for a thermogenetic manipulation of individual neurons using IR light., Current approaches to thermogenetic manipulation of neuronal activity lack sufficient spatiotemporal resolution. Here the authors show that neurons expressing snake TRPA1 channels are activated at high temporal resolution with IR light and this technique can be used to elicit behaviour in zebrafish larvae.

Hepatitis E virus (HEV) has emerged as a cause of chronic hepatitis among immunocompromised patients. Molecular assays have become important tools for the diagnosis and management of these chronically infected patients. A real-time reverse transcription-quantitative PCR (RT-qPCR) assay utilizing Pleiades probe chemistry and an RNA internal control for the simultaneous detection and quantification of HEV RNA in human serum was developed based on an adaptation of a previously described and broadly reactive primer set targeting the overlapping open reading frame 2/3 (ORF2/3) nucleotide sequence of HEV. A chimeric bovine viral diarrhea virus construct containing an HEV RNA insert (SynTura HEV) was developed, value assigned with the first World Health Organization (WHO) international standard for HEV RNA (code 6329/10), and used to prepare working assay calibrators and controls, which supported an assay quantification range of 100 to 5,000,000 IU/ml. The analytical sensitivity (95% detection rate) of this assay was 25.2 IU/ml (95% confidence interval [CI], 19.2 to 44.1 IU/ml). The assay successfully amplified 16 different HEV sequences with significant nucleotide mismatching in primer/probe binding regions, while evaluation of a WHO international reference panel for HEV genotypes (code 8578/13) showed viral load results falling within the result ranges generated by WHO collaborative study participants for all panel members (genotypes 1 to 4). Broadly reactive RT-qPCR primers targeting HEV ORF2/3 were successfully adapted for use in an assay based on Pleiades probe chemistry. The availability of secondary standards calibrated to the WHO HEV international standard can improve the standardization and performance of assays for the detection and quantification of HEV RNA.

The osteogenic, cytotoxic, and antibacterial activities of polysaccharide (PS-SC) and flavonoid (F-SC) fractions isolated from the leaves extract of Saussurea controversa were studied in vitro. F-SC consists of the five quercetin glycosides in the ratio 2:8:10:1:4, which were isolated from the leaves extract of S. controversa and have been characterized previously. PS-SC was first isolated from the leaves extract of S. controversa and has been described. PS-SC consists in 30 compounds is characterized by a high degree of heterogeneity with a heterogeneity index of 19.74. The Mw and Mn of PS-SC were 108.6 and 5.5 kDa, respectively. Structural fragments are represented by galactose, arabinose, xylose, glucose, uronic acids, mannose, and rhamnose in a 10.1:3.3:2.2:2.1:1.7:0.9:0.5 molar ratio. F-SC as compared with PS-SC showed in vitro microbicidal (50 g/L) and better bacteriostatic (6.25 g/L versus 25 g/L of PS-SC) effects against the 24-h growth of Staphylococcus aureus strain 209 P and a 21-day absence of cytotoxicity on human adipose-derived multipotent mesenchymal stromal cells (hAMMSCs). Both fractions (PS-SC > F-SC) at doses of 10–50 mg/L stimulated differentiation of hAMMSCs into secreting osteoblasts accompanied by local mineralization of extracellular matrix. These fractions of S. controversa and especially F-SC, might be promising peroral drugs in the complex treatment of bone fractures and for prophylaxis of their infectious complications.

Genetically encoded biosensors based on fluorescent proteins (FPs) are a reliable tool for studying the various biological processes in living systems. The circular permutation of single FPs led to the development of an extensive class of biosensors that allow the monitoring of many intracellular events. In circularly permuted FPs (cpFPs), the original N- and C-termini are fused using a peptide linker, while new termini are formed near the chromophore. Such a structure imparts greater mobility to the FP than that of the native variant, allowing greater lability of the spectral characteristics. One of the common principles of creating genetically encoded biosensors is based on the integration of a cpFP into a flexible region of a sensory domain or between two interacting domains, which are selected according to certain characteristics. Conformational rearrangements of the sensory domain associated with ligand interaction or changes in the cellular parameter are transferred to the cpFP, changing the chromophore environment. In this review, we highlight the basic principles of such sensors, the history of their creation, and a complete classification of the available biosensors.

Reactive oxygen species (ROS) regulate both normal cell functions by activating a number of enzymatic cascades and pathological processes in many diseases by inducing oxidative stress. For many years since the discovery of ROS in biological systems there were no adequate methods of detection and quantification of these molecules inside the living cells. We developed the first genetically encoded fluorescent indicator for intracellular detection of hydrogen peroxide, HyPer, that can be used for imaging of H

Hydrogen peroxide (H2O2) plays an important role in modulating cell signaling and homeostasis in live organisms. The HyPer family of genetically encoded indicators allows the visualization of H2O2 dynamics in live cells within a limited field of view. The visualization of H2O2 within a whole organism with a single cell resolution would benefit from a slowly reducible fluorescent indicator that integrates the H2O2 concentration over desired time scales. This would enable post hoc optical readouts in chemically fixed samples. Herein, we report the development and characterization of NeonOxIrr, a genetically encoded green fluorescent indicator, which rapidly increases fluorescence brightness upon reaction with H2O2, but has a low reduction rate. NeonOxIrr is composed of circularly permutated mNeonGreen fluorescent protein fused to the truncated OxyR transcription factor isolated from E. coli. When compared in vitro to a standard in the field, HyPer3 indicator, NeonOxIrr showed 5.9-fold higher brightness, 15-fold faster oxidation rate, 5.9-fold faster chromophore maturation, similar intensiometric contrast (2.8-fold), 2-fold lower photostability, and significantly higher pH stability both in reduced (pKa of 5.9 vs. &ge, 7.6) and oxidized states (pKa of 5.9 vs.&ge, 7.9). When expressed in the cytosol of HEK293T cells, NeonOxIrr demonstrated a 2.3-fold dynamic range in response to H2O2 and a 44 min reduction half-time, which were 1.4-fold lower and 7.6-fold longer than those for HyPer3. We also demonstrated and characterized the NeonOxIrr response to H2O2 when the sensor was targeted to the matrix and intermembrane space of the mitochondria, nucleus, cell membranes, peroxisomes, Golgi complex, and endoplasmic reticulum of HEK293T cells. NeonOxIrr could reveal endogenous reactive oxygen species (ROS) production in HeLa cells induced with staurosporine but not with thapsigargin or epidermal growth factor. In contrast to HyPer3, NeonOxIrr could visualize optogenetically produced ROS in HEK293T cells. In neuronal cultures, NeonOxIrr preserved its high 3.2-fold dynamic range to H2O2 and slow 198 min reduction half-time. We also demonstrated in HeLa cells that NeonOxIrr preserves a 1.7-fold ex vivo dynamic range to H2O2 upon alkylation with N-ethylmaleimide followed by paraformaldehyde fixation. The same alkylation-fixation procedure in the presence of NP-40 detergent allowed ex vivo detection of H2O2 with 1.5-fold contrast in neuronal cultures and in the cortex of the mouse brain. The slowly reducible H2O2 indicator NeonOxIrr can be used for both the in vivo and ex vivo visualization of ROS. Expanding the family of fixable indicators may be a promising strategy to visualize biological processes at a single cell resolution within an entire organism.

To compare fecal calprotectin (FC) concentration with laboratory and diagnostic methods in patients with inflammatory bowel diseases (IBD).The level of FC was measured in 110 patients with established IBD. Crohn diseases (CD) was diagnosed in 50 patients, ileocolitis - in 38 and terminal ileitis in 12 individuals. Ulcerative colitis (UC) was diagnosed in 60 patients, total colitis in 35, left-side colitis in 21 and 4 patients have proctitis. Laboratory data include measurement of FC, leukocytes, erythrocyte sedimentation rate (ESR), C reactive protein (CRP), fecal occult blood. All patients underwent colonoileoscopy (CIS) at the start of disease flare and after 12 weeks of treatment.We found linear correlation between level of FCP and endoscopic activity of CD, analyzing FCP level and endoscopic activity of CD before (during disease flare) and after 12 weeks treatment (r=0.66, plt;0.001). Linear correlation between FCP and SES-CD sustained after 12 weeks of treatment (r=0.77, plt;0.001). We revealed correlation between FCP concentration. And CRP level (r=0.59, plt;0.05). The linear correlation was detected between FCP and endoscopic activity of UC (r=0.88, plt;0.001) before the treatment. After 12 weeks of treatment linear correlation was shown between FCP and Meyo scale (r=0.73, plt;0.001). IBD patients with FCP more than 200 mcg/g have high risk of disease reccurence in short-term period of time (HR - 8.33; 95% CI 2.05-33.8; χ2 - 11.85; plt;0.001) and (HR - 2.7; 95% CI 1.1-6.6; χ2 - 5.3; plt;0.05), accordingly.Increased FCP level indicates poor effectiveness of treatment and high risk of reccurence. The level of FCP correlates strongly with recent laboratory and diagnostic indices of activity and enables to determine patients with high risk of reccurence. Thus, thorough monitoring, including additional procedures, contributes to just-in-time treatment modification.

The main challenge in electron external beam radiation therapy with clinical accelerators is the absence of integrated systems to form irregular fields. The current approach to provide conformal irradiation is to use additional metallic shaping blocks, with inefficient and expensive workflows. This work presents a simple method to form therapeutic electron fields using 3D printed samples. These samples are manufactured by fused deposition modeling, which can affect crucial properties, such as material homogeneity, due to the presence of residual air-filled cavities. The applicability of this method was therefore investigated with a set of experiments and Monte Carlo simulations aimed at determining the electron depth dose distribution in polymer materials. The results show that therapeutic electron beams with energies 6–20 MeV can be effectively absorbed using these polymeric samples. The model developed in this study provides a way to assess the dose distribution in such materials and to calculate the appropriate thickness of polymer samples for therapeutic electron beam formation. It is shown that for total absorption of 6 MeV electron beams the material thickness should be at least 4 cm, while this value should be at least 8 cm for 12 MeV and 11 cm for 20 MeV, respectively. The results can be used to further develop 3D printing procedures for medical electron beam profile formation, allowing the creation of a collimator or absorber with patient-specific configuration using rapid prototyping systems, thus contributing to improve the accuracy of dose delivery in electron radiotherapy within a short manufacturing time.

Papillary carcinoma is the most commonly diagnosed form of well-differentiated thyroid cancer that is generally characterized by a favorable prognosis. However, a number of relatively rare variants of this tumor, such as papillary carcinoma of high cells, papillary carcinoma of columnar cells, a diffuse sclerosing variant and recently described cancer of shoe nail cell type, are characterized by a less favorable clinical course, a high frequency of distant metastasis, and relatively low overall and relapse-free survival rates. In this connection, it is important to recognize these options at the stage of a primary morphological study. This review of the literature considers the morphological, clinical and molecular genetic features of the above variants of papillary thyroid carcinoma.Папиллярный рак является наиболее часто диагностируемой формой высокодифференцированного рака щитовидной железы и, как правило, характеризуется благоприятным прогнозом. Однако ряд сравнительно редко встречающихся вариантов данной опухоли, таких как папиллярный рак из высоких клеток, папиллярный рак из столбчатых клеток, диффузно-склерозирующий вариант и недавно описанный рак из клеток типа 'сапожных гвоздей', характеризуются менее благоприятным клиническим течением, высокой частотой отдаленного метастазирования и относительно низким уровнем общей и безрецидивной выживаемости. В связи с этим данные варианты важно распознавать на этапе первичного морфологического исследования. В настоящем обзоре литературы рассмотрены морфологические, клинические и молекулярно-генетические особенности вышеперечисленных прогностически неблагоприятных вариантов папиллярного рака щитовидной железы.

Collagen is one of the most promising materials for 3D bioprinting because of its distinguished biocompatibility. Cell-laden constructs made of pure collagen with or without incorporated growth supplements support engineered constructs persistence in culture and are perfectly suitable for grafting. The limiting factor for direct 3D collagen printing was poor printability of collagen solutions, especially admixed with cells or tissue spheroids. In our study, we showed that concentrated solutions of native collagen branded Viscoll were effective as bioinks with high fidelity performance. Viscoll containing 20, 30, or 40 mg/ml collagen were used for direct extrusion 3D bioprinting to form scaffolds appropriate to support spatial arrangement of tissue spheroids into rigid patterns with resolution of 0.5 mm in details. Incorporated cells demonstrated sufficient viability. Associated rheological study showed that good printability of the collagen solutions correlates with their increased storage modulus value, notably exceeding the loss modulus value. The proper combination of these physical parameters could become technological criteria for manufacturing various collagen bioinks for 3D bioprinting.

Prostatitis is considered as heterogeneous group of the diseases attracting broad interest of researchers worldwide. The acute and chronic stages of the process, as particular categories of bacterial prostatitis, remain in the realities of modern practical and scientific urology with lively discussions. Despite a large number of domestic and international publications, consensus on many topical issues of bacterial prostatitis is currently not achieved. The true incidence and prevalence of bacterial prostatitis in various regions of the world has not been finally determined. The exhaustive data concerning the etiological structure, mechanisms of development and persistence of the inflammatory process in the prostate tissue are not presented. At the same time, new risk factors that can influence the development and progress of these diseases are identified and discussed. Researchers review algorithms for examining patients, obligate spectrum of diagnostic methods, as well as the desirability of using the newest investment means necessary for the successful verification of the diagnosis taking into account the development of modern medical technologies. This review describes in detail the various aspects of the epidemiology and etiology of bacterial prostatitis, based on an analysis of major literature sources in conjunction with its own scientific facts.