Specific inhibition of acetylcholinesterase as an approach to decrease muscarinic side effects during myasthenia gravis treatment

© 2017 The Author(s). Non-selective inhibitors of cholinesterases (ChEs) are clinically used for treatment of myasthenia gravis (MG). While being generally safe, they cause numerous adverse effects including induction of hyperactivity of urinary bladder and intestines affecting quality of patients life. In this study we have compared two ChEs inhibitors, a newly synthesized compound C547 and clinically used pyridostigmine bromide, by their efficiency to reduce muscle weakness symptoms and ability to activate contractions of urinary bladder in a rat model of autoimmune MG. We found that at dose effectively reducing MG symptoms, C547 did not affect activity of rat urinary bladder. In contrast, at equipotent dose, pyridostigmine caused a significant increase in tonus and force of spontaneous contractions of bladder wall. We also found that this profile of ChEs inhibitors translates into the preparation of human urinary bladder. The difference in action observed for C547 and pyridostigmine we attribute to a high level of pharmacological selectivity of C547 in inhibiting acetylcholinesterase as compared to butyrylcholinesterase. These results raise reasonable hope that selective acetylcholinesterase inhibitors should show efficacy in treating MG in human patients with a significant reduction in adverse effects related to hyperactivation of smooth muscles

Assessment of orthologous splicing isoforms in human and mouse orthologous genes

<p>Abstract</p> <p>Background</p> <p>Recent discoveries have highlighted the fact that alternative splicing and alternative transcripts are the rule, rather than the exception, in metazoan genes. Since multiple transcript and protein variants expressed by the same gene are, by definition, structurally distinct and need not to be functionally equivalent, the concept of gene orthology should be extended to the transcript level in order to describe evolutionary relationships between structurally similar transcript variants. In other words, the identification of true orthology relationships between gene products now should progress beyond primary sequence and "splicing orthology", consisting in ancestrally shared exon-intron structures, is required to define orthologous isoforms at transcript level.</p> <p>Results</p> <p>As a starting step in this direction, in this work we performed a large scale human- mouse gene comparison with a twofold goal: first, to assess if and to which extent traditional gene annotations such as RefSeq capture genuine splicing orthology; second, to provide a more detailed annotation and quantification of true human-mouse orthologous transcripts defined as transcripts of orthologous genes exhibiting the same splicing patterns.</p> <p>Conclusions</p> <p>We observed an identical exon/intron structure for 32% of human and mouse orthologous genes. This figure increases to 87% using less stringent criteria for gene structure similarity, thus implying that for about 13% of the human RefSeq annotated genes (and about 25% of the corresponding transcripts) we could not identify any mouse transcript showing sufficient similarity to be confidently assigned as a splicing ortholog. Our data suggest that current gene and transcript data may still be rather incomplete - with several splicing variants still unknown. The observation that alternative splicing produces large numbers of alternative transcripts and proteins, some of them conserved across species and others truly species-specific, suggests that, still maintaining the conventional definition of gene orthology, a new concept of "splicing orthology" can be defined at transcript level.</p

Медикаментозная гипокоагуляция: результаты экстренного хирургического лечения

Introduction. The drug-induced hemorrhagic syndrome in surgical patients has become a serious problem of contemporary surgery, complicating the choice of treatment tactics and often leading to death. The present study is aimed at optimising emergency surgical treatment of acute cholecystitis and strangulated hernia in patients receiving warfarin.Material and methods. The study involved 18 patients with hypocoagulant state caused by warfarin (the main group). The control group (10 patients) consisted of patients taking no drugs affecting the coagulation profile. The study was conducted during 2013-2019. The range of diseases included acute cholecystitis and strangulated hernia. The scope of surgery was presented by laparoscopic cholecystectomy and cholecystectomy using minimally invasive technologies. In patients with strangulated hernias, prosthetic tension-free hernioplasty and local tissue rearrangement were used. In the main group, the INR values comprised 8.7 ± 2.8 and 8.4 ± 0.8 in the case of laparoscopic cholecystectomy and short-scar incision, respectively. In patients with strangulated hernias, the pre-surgery INR indices were 8.1 ± 2.7 and 8.0 ± 1.5 with local tissue rearrangement and prosthetic hernioplasty, respectively. The examination included: complete blood count, common urine analysis, US, endoscopic examination, etc.; in the setting of hemorrhagic syndrome, coagulogram, INR and others were added.Results and discussion. In patients with hypocoagulation syndrome, no significant differences were observed in coagu-logram parameters of the pre- and postoperative period in spite of the corrective therapy by quarantine fresh-frozen plasma. The patients with acute cholecystitis receiving warfarin demonstrated the contact bleeding of the liver and longterm serous-hemorrhagic discharge through the drains. However, the rate of hemorrhage in the group of patients with short-scar incision cholecystectomy was observed to be significantly higher as compared to the laparoscopy-operated group. In patients with strangulated hernias, in cases of drainage of the postoperative wound, a long-term serous-hemorrhagic discharge was observed.Conclusion. Laparoscopic cholecystectomy is the operation of choice in patients receiving warfarin and developing acute cholecystitis. However, in case of strangulation in patients with hernias, the optimal operation consists in local tissue rearrangement.Введение. Геморрагический синдром у хирургических больных, обусловленный приемом медикаментозных препаратов, стал серьезной проблемой современной хирургии, осложняющей выбор тактики лечения и нередко приводящей к летальному исходу. Цель исследования — оптимизировать экстренное хирургическое лечение острого холецистита и ущемленной грыжи у больных, принимающих варфарин.Материал и методы. Исследование включало 18 пациентов, гипокоагуляционое состояние которых было обусловлено приемом варфарина (основная группа). Контрольную группу (10 пациентов) составили больные, не принимавшие препараты, влияющие на коагуляционный профиль. Сроки исследования — 2013-2019 гг. Спектр заболеваний включал острый холецистит и ущемленную грыжу. Объем оперативного вмешательства включал лапароскопическую холецистэктомию и холецистэктомию с применением минимально инвазивных технологий. У больных с ущемленными грыжами применялась протезирующая ненатяжная герниопластика и пластика местными тканями. Показатели МНО в основной группе больных составили 8,7 ± 2,8 в случае применения лапароскопической холецистэктомии и 8,4 ± 0,8 при операции через мини-доступ. У больных с ущемленными грыжами показатели МНО до операции составили — 8,1 ± 2,7 при пластике местными тканями, у больных с протезирующей герниопластикой 8,0 ± 1,5. Обследование включало общий анализ крови, общий анализ мочи, УЗИ, эндоскопическое исследование и др., при геморрагическом синдроме — коагулограмма, МНО и пр.Результаты и обсуждение. Показатели коагулограммы у больных с гипокоагуляционным синдромом достоверно не отличались в пред- и послеоперационном периоде, несмотря на проведенную корригирующую терапию с применением свежезамороженной плазмы. Установлено, что у больных с острым холециститом, принимавших варфарин, наблюдается контактная кровоточивость печени, длительное истечение серозно-геморрагического отделяемого по дренажам. Однако дебит геморрагии в группе больных с холецистэктомией из мини-доступа был существенно больше в сравнении с группой, оперированной с использованием лапароскопии. У больных с ущемленными грыжами в случаях дренирования послеоперационной раны наблюдалось длительное истечение серозно-геморрагического отделяемого.Заключение. У больных, принимающих варфарин, при развитии острого холецистита предпочтительной операцией является лапароскопическая холецистэктомия. В случае возникновения ущемления у больных с грыжами оптимальной операцией является пластика местными тканями

ВЫСОКИЙ УРОВЕНЬ ГЛИКОЗАМИНОГЛИКАНОВ СЫВОРОТКИ КРОВИ КАК НЕЗАВИСИМЫЙ ПРЕДИКТОР РАЗВИТИЯ СПАЕЧНОЙ БОЛЕЗНИ БРЮШИНЫ

Peritoneal commissures is the phenomenon of adhesions formation with apparent symptom group of violations in functioning of the gastrointestinal tract and small pelvis. Unlike other postoperative complications adhesive disease is characterized by high risk for life manifestations in the form of intestinal obstruction, infertility and chronic pain. It is known that in the process of restoring the integrity of the peritoneum the decisive role is given to makrophages, peritoneal immune system, processes of angiogenesis, increasing fibroblasts and collagen production. However, the biochemical processes taking place in the intercellular matrix of connective tissue, particularly, the exchange of glycosaminoglycans in the formation of peritoneal adhesions have not been studied. In this context, the main objective of this work was to study levels of glycosaminoglycans of the blood in the development of adhesive disease and justification of the use of indicators to exchange connective tissue as additional prognostic criteria for the development of postoperative adhesions. An open prospective randomized trial analyzed the data of 67 patients that were admitted for the planned reconstructive surgery on the anterior abdominal wall to eliminate postoperative ventral hernia, at the surgery department of State Clinical Hospital No. 8, city ofUFAduring the period of 2005-2008. The severity of adhesions was assessed and contrasted to the results of determination of glycosaminoglycans in blood serum. The total content of glycosaminoglycans, hyaluronic acid, chondroitin sulfates and heparan sulfates was determined. Postoperative monitoring stages of the studied indicators -3, 5 and 7 day after herniotomy. The group of patients with adhesive peritoneum desease showed total content of glycosaminoglycans by 19.2% (p &lt; 0.0001) higher than reference values. It should be noted that there was continuous growth of all factions of glycosaminoglycans during all seven days of the postoperative monitoring. The growth of the overall content of the glycosaminoglycans made 261% (p &lt; 0.0001) compared with the control group. Method of ROC-analysis established that the area under the ROC curve is a common glycosaminoglycans is equal 0.824 ± 0.055, 95% confidence interval-0.906 0.711. Sensitivity-70.4% (54.8 -86.0), specificity is 97.5% (82.4 -98.6). The optimum point for the division amounted to 30 μmol/l. Positive predicting level amounted to 96.8%, negative 75.1%. Thus, the assessment of the level of glycosaminoglycans can serve as an additional prognostic test to diagnose disturbance in intercellular matrix that can be used in the diagnosis of commissural process and in monitoring the effectiveness of preventive measures. Спаечная болезнь – это явление образования спаек с выраженным симптомокомплексом нарушения функционирования органов желудочно-кишечного тракта и малого таза. В отличие от других послеоперационных осложнений спаечная болезнь характеризуется пожизненным высоким риском проявлений кишечной непроходимости, бесплодия и хронических болей. Известно, что в процессе восстановления целостности брюшины решающую роль играют макрофаги, иммунная система брюшины, процессы ангиогенеза, увеличение продукции фибробластов и коллагена. Однако биохимические процессы, происходящие в межклеточном матриксе соеди-нительной ткани, в частности обмен гликозаминогликанов при формировании брюшинных спаек, не изучены. В этой связи основной целью данной работы являлось исследование уровней гликозаминогликанов крови при развитии спаечной болезни брюшины и обоснование использования показателей обмена соединительной ткани в качестве дополнительных прогностических критериев развития послеоперационных спаек. В рамках открытого проспективного рандомизированного исследования мы проанализировали данные 67 пациентов, поступавших для планового проведения реконструктивной операции на передней брюшной стенке с целью устранения послеоперационной вентральной грыжи в хирургическое отделение ГКБ №8 г. Уфы в период 2005-2008 гг. Проводили оценку выраженности спаечного процесса и сопоставляли с результатами определения гликозаминогликанов сыворотки крови. Определяли общее содержание гликозаминогликанов, гиалуроновой кислоты, хондроитинсульфатов и гепарансульфатов. Этапы послеоперационного мониторинга изучаемых показателей – 3-и, 5и 7-е сутки после грыжесечения. Установлено, что в группе пациентов со спаечной болезнью брюшины общее содержание гликозаминогликанов на 19,2% (р&lt;0,0001) выше контрольных значений. Следует отметить, что регистрировался непрерывный рост всех фракций гликозаминогликанов на протяжении всех семи суток послеоперационного мониторинга. Рост общего содержания ГАГ составил 261% (р&lt;0,0001) в сравнении с группой контроля. Методом ROC-анализа установлено, что площадь под ROC-кривой для уровня общего ГАГ оказалась равной 0,824±0,055, 95% доверительный интервал составил 0,711 0,906, чувствительность – 70,4% (54,8-86,0), специфичность – 97,5% (82,4-98,6). Оптимальная точка разделения составила 30 мкмоль/л. Положительный предсказывающий уровень составил 96,8%, отрицательный – 75,1%. Таким образом, оценка уровня гликозаминогликанов может являться дополнительным прогностическим тестом диагностики нарушения структуры межклеточного матрикса, что может быть использовано и в диагностике спаечного процесса, и в контроле эффективности профилактических мероприятий.

Острая тонкокишечная непроходимость, вызванная эктопированным участком поджелудочной железы. Клинический случай

Introduction. An ectopic pancreas is an abnormality in which pancreatic tissue has grown outside its normal location with its own blood supply and ducts while having no anatomical, vascular or innervation connections with the pancreas situated normally. The accessory pancreas is the most frequent congenital disorder of this gland. The pancreatic tissue can be found in the walls of stomach, intestine, gallbladder, Meckel’s diverticulum, the liver, spleen and, seldom, in other organs having no contact with the pancreas. Currently the ectopic pancreas incidence is on the rise and amounts up to 0.2% on average in abdominal surgeries and is reported in 0.5–13% of autopsy cases. Most frequently the aberrant pancreas is located in the gastroduodenal zone (63–70% of all the heterotopic pancreas cases); most often it is found in the pylorus and antrum (85–95% of all the gastric ectopic pancreas cases).Materials and methods. This paper presents a case of a successful treatment of a 39 year old male with an acute small intestinal obstruction caused by ectopic pancreatic tissue in the intestinal wall. The patient received a diagnostic laparoscopy and a mid-midline laparotomy with the wedge resection of the small intestine.Results.The postoperative period was recorded as uneventful; the patient was discharged home on day nine following the surgery after removal of cutaneous sutures. The pathology of the small intestinal fragment with the polyp-like neoplasm identified heterotypic pancreatic foci (with acini and ducts) located between muscular bands, with extensive areas of haemorrhaging and necrosis.Conclusion.The diagnosis of the ectopic pancreas tissue in intestinal wall is an extremely complex issue in abdominal surgery; this diagnosis is normally made only when complications occur. Pathomorphological verification is of paramount importance to make the final diagnosis of this disease, making it possible to identify correctly the cause and mechanism of the development of an acute surgical disorder.Введение. Эктопия поджелудочной железы — это аномалия, связанная с необычным расположением панкреатической ткани с собственным кровоснабжением и протоками. Она не имеет анатомического, сосудистого и нервного контакта с нормально расположенной поджелудочной железой. Добавочная поджелудочная железа (ПЖ), самый частый порок развития железы, который заключен в гетеротопии ее ткани в стенку желудка, кишечника, желчного пузыря, дивертикул Меккеля, печень, селезенку и, значительно реже, в другие органы без связи с основной поджелудочной железой. В настоящее время частота встречаемости гетеротопии поджелудочной железы значительно увеличилась и составляет в среднем до 0,2 % при оперативных вмешательствах на органах брюшной полости и 0,5–13 % случаев при аутопсиях. Наиболее часто аберрантная ПЖ локализована в гастродуоденальной зоне (63–70 % от всех случаев гетеротопии ПЖ) с преимущественным расположением в антральном и пилорическом отделах желудка (85–95 % от всех случаев гетеротопии ПЖ в желудке).Материалы и методы. В статье представлен клинический пример успешного лечения 39-летнего мужчины с острой тонкокишечной непроходимостью, вызванной эктопированной тканью поджелудочной железы в стенку тонкой кишки. Пациенту была выполнена диагностическая лапароскопия, произведена средне-срединная лапаротомия с клиновидной резекцией тонкой кишки.Результаты. Послеоперационный период гладкий, больной выписан на девятые сутки после хирургического вмешательства после снятия кожных швов. По результатам гистологического исследования: фрагмент тонкой кишки и полиповидное образование, представленное гетеротопированными участками поджелудочной железы, состоящими из ацинусов и проток, располагающихся между мышечными тяжами, с обширными кровоизлияниями и некрозами.Заключение. Диагностика эктопированной ткани поджелудочной железы в тонкую кишку является крайне сложной проблемой в абдоминальной хирургии и выявляется в большинстве случаев при развитии осложнений. Для окончательной диагностики данного заболевания имеет огромное значение патоморфологическая верификация, которая позволяет правильно пределить причину и механизм развития острого хирургического заболевани

Diversity of Protein and mRNA Forms of Mammalian Methionine Sulfoxide Reductase B1 Due to Intronization and Protein Processing

Background: Methionine sulfoxide reductases (Msrs) are repair enzymes that protect proteins from oxidative stress by catalyzing stereospecific reduction of oxidized methionine residues. MsrB1 is a selenocysteine-containing cytosolic/nuclear Msr with high expression in liver and kidney. Principal Findings: Here, we identified differences in MsrB1 gene structure among mammals. Human MsrB1 gene consists of four, whereas the corresponding mouse gene of five exons, due to occurrence of an additional intron that flanks the stop signal and covers a large part of the 3′-UTR. This intron evolved in a subset of rodents through intronization of exonic sequences, whereas the human gene structure represents the ancestral form. In mice, both splice forms were detected in liver, kidney, brain and heart with the five-exon form being the major form. We found that both mRNA forms were translated and supported efficient selenocysteine insertion into MsrB1. In addition, MsrB1 occurs in two protein forms that migrate as 14 and 5 kDa proteins. We found that each mRNA splice form generated both protein forms. The abundance of the 5 kDa form was not influenced by protease inhibitors, replacement of selenocysteine in the active site or mutation of amino acids in the cleavage site. However, mutation of cysteines that coordinate a structural zinc decreased the levels of 5 and 14 kDa forms, suggesting importance of protein structure for biosynthesis and/stability of these forms. Conclusions: This study characterized unexpected diversity of protein and mRNA forms of mammalian selenoprotein MsrB1

Search for Specific Biomarkers of IFNβ Bioactivity in Patients with Multiple Sclerosis

Myxovirus A (MxA), a protein encoded by the MX1 gene with antiviral activity, has proven to be a sensitive measure of IFNβ bioactivity in multiple sclerosis (MS). However, the use of MxA as a biomarker of IFNβ bioactivity has been criticized for the lack of evidence of its role on disease pathogenesis and the clinical response to IFNβ. Here, we aimed to identify specific biomarkers of IFNβ bioactivity in order to compare their gene expression induction by type I IFNs with the MxA, and to investigate their potential role in MS pathogenesis. Gene expression microarrays were performed in PBMC from MS patients who developed neutralizing antibodies (NAB) to IFNβ at 12 and/or 24 months of treatment and patients who remained NAB negative. Nine genes followed patterns in gene expression over time similar to the MX1, which was considered the gold standard gene, and were selected for further experiments: IFI6, IFI27, IFI44L, IFIT1, HERC5, LY6E, RSAD2, SIGLEC1, and USP18. In vitro experiments in PBMC from healthy controls revealed specific induction of selected biomarkers by IFNβ but not IFNγ, and several markers, in particular USP18 and HERC5, were shown to be significantly induced at lower IFNβ concentrations and more selective than the MX1 as biomarkers of IFNβ bioactivity. In addition, USP18 expression was deficient in MS patients compared with healthy controls (p = 0.0004). We propose specific biomarkers that may be considered in addition to the MxA to evaluate IFNβ bioactivity, and to further explore their implication in MS pathogenesis

Crowdsourced estimation of cognitive decline and resilience in Alzheimer's disease

Identifying accurate biomarkers of cognitive decline is essential for advancing early diagnosis and prevention therapies in Alzheimer's disease. The Alzheimer's disease DREAM Challenge was designed as a computational crowdsourced project to benchmark the current state-of-the-art in predicting cognitive outcomes in Alzheimer's disease based on high dimensional, publicly available genetic and structural imaging data. This meta-analysis failed to identify a meaningful predictor developed from either data modality, suggesting that alternate approaches should be considered for prediction of cognitive performance

Predicting Functional Alternative Splicing by Measuring RNA Selection Pressure from Multigenome Alignments

High-throughput methods such as EST sequencing, microarrays and deep sequencing have identified large numbers of alternative splicing (AS) events, but studies have shown that only a subset of these may be functional. Here we report a sensitive bioinformatics approach that identifies exons with evidence of a strong RNA selection pressure ratio (RSPR) —i.e., evolutionary selection against mutations that change only the mRNA sequence while leaving the protein sequence unchanged—measured across an entire evolutionary family, which greatly amplifies its predictive power. Using the UCSC 28 vertebrate genome alignment, this approach correctly predicted half to three-quarters of AS exons that are known binding targets of the NOVA splicing regulatory factor, and predicted 345 strongly selected alternative splicing events in human, and 262 in mouse. These predictions were strongly validated by several experimental criteria of functional AS such as independent detection of the same AS event in other species, reading frame-preservation, and experimental evidence of tissue-specific regulation: 75% (15/20) of a sample of high-RSPR exons displayed tissue specific regulation in a panel of ten tissues, vs. only 20% (4/20) among a sample of low-RSPR exons. These data suggest that RSPR can identify exons with functionally important splicing regulation, and provides biologists with a dataset of over 600 such exons. We present several case studies, including both well-studied examples (GRIN1) and novel examples (EXOC7). These data also show that RSPR strongly outperforms other approaches such as standard sequence conservation (which fails to distinguish amino acid selection pressure from RNA selection pressure), or pairwise genome comparison (which lacks adequate statistical power for predicting individual exons)

Functional annotation of human long noncoding RNAs via molecular phenotyping

Long noncoding RNAs (lncRNAs) constitute the majority of transcripts in the mammalian genomes, and yet, their functions remain largely unknown. As part of the FANTOM6 project, we systematically knocked down the expression of 285 lncRNAs in human dermal fibroblasts and quantified cellular growth, morphological changes, and transcriptomic responses using Capped Analysis of Gene Expression (CAGE). Antisense oligonucleotides targeting the same lncRNAs exhibited global concordance, and the molecular phenotype, measured by CAGE, recapitulated the observed cellular phenotypes while providing additional insights on the affected genes and pathways. Here, we disseminate the largest-todate lncRNA knockdown data set with molecular phenotyping (over 1000 CAGE deep-sequencing libraries) for further exploration and highlight functional roles for ZNF213-AS1 and lnc-KHDC3L-2.Peer reviewe