Equivalence of Conventionally-Derived and Parthenote-Derived Human Embryonic Stem Cells

As human embryonic stem cell (hESC) lines can be derived via multiple means, it is important to determine particular characteristics of individual lines that may dictate the applications to which they are best suited. The objective of this work was to determine points of equivalence and differences between conventionally-derived hESC and parthenote-derived hESC lines (phESC) in the undifferentiated state and during neural differentiation.hESC and phESC were exposed to the same expansion conditions and subsequent neural and retinal pigmented epithelium (RPE) differentiation protocols. Growth rates and gross morphology were recorded during expansion. RTPCR for developmentally relevant genes and global DNA methylation profiling were used to compare gene expression and epigenetic characteristics. Parthenote lines proliferated more slowly than conventional hESC lines and yielded lower quantities of less mature differentiated cells in a neural progenitor cell (NPC) differentiation protocol. However, the cell lines performed similarly in a RPE differentiation protocol. The DNA methylation analysis showed similar general profiles, but the two cell types differed in methylation of imprinted genes. There were no major differences in gene expression between the lines before differentiation, but when differentiated into NPCs, the two cell types differed in expression of extracellular matrix (ECM) genes.These data show that hESC and phESC are similar in the undifferentiated state, and both cell types are capable of differentiation along neural lineages. The differences between the cell types, in proliferation and extent of differentiation, may be linked, in part, to the observed differences in ECM synthesis and methylation of imprinted genes

How ligand binds to the type 1 insulin-like growth factor receptor

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Пресепсин в диагностике нозокомиальной инфекции центральной нервной системы

ABSTRACT Introduction Nosocomial infection of the central nervous system (NI-CNS) is a serious complication in neurocritical patients that leads to deterioration of patient’s condition, worsening of outcomes and increased cost of treatment. The timely diagnosis of NI-CNS is a relevant problem and the search for new reliable markers of NI-CNS is an important issue.MATERIAL AND METHODS The prospective observational study consisted of two parts. The aim of the frst part was to defne normal ranges of cerebral spinal presepsin (CSF PSP). The aim of the second part was investigation of CSF PSP in neurocritical patients. We studied CSF sampling obtained during spinal anesthesia for elective urologic surgery in order to defne the normal CSF PSP. The following data was collected in neurocritical patients: CSF cell count, glucose, lactate, PSP, microbiological tests, polymerase chain reaction (PCR), when it was possible. Blood tests included complete blood count, C-reactive protein (CRP), procalcitonin (PCT), PSP. IBM SPSS Statistics (version 23.0) was used for statistical analysis.RESULTS Fifteen CSF samplings were obtained for investigation of normal CSF PSP ranges, which was 50–100 pg/ml. Nineteen neurocritical patients were included. Sixty-three pairs of CSF and blood samplings were obtained. All pairs were divided into the 4 groups in accordance with presence/absence of NI-CNS or systemic infection. In cases without both NI-CNS and systemic infection (group 4) CSF PSP was 406±203.1 pg/ml. In cases without NI-CNS and with systemic infection (group 2) CSF PSP was 614.9±315 pg/ml. In cases with NI-CNS and without systemic infection (group 3) CSF PSP was 547.8±264.3 pg/ml. In cases with both NI-CNS and systemic infection (group 1) CSF PSP was 731.1±389.7 pg/ml. The ROC analysis showed that in neurocritical patients without systemic infection CSF PSP 537 pg/ml meant NI-CNS with sensitivity 68.8% and specifcity 85.7%.CONCLUSION The normal value of the CSF PSP is 50-100 pg/ml. CSF PSP more than 537 pg/ml in neurocritical patients without systemic infection meant NI-CNS with 688% sensitivity and 857% specifcity. CSF PSP may be used for diagnosing NI-CNS in neurocritical patients as an additional marker only. CSF may be used as an additional diagnostic criterion, but further research is needed.ВВЕДЕНИЕ Нозокомиальная инфекция центральной нервной системы (НИ ЦНС) является тяжелым осложнением, приводящим к ухудшению состояния, удлинению продолжительности лечения и ухудшению исходов заболевания у нейрореанимационных пациентов. Ранняя диагностика НИ ЦНС является актуальной клинической задачей, а поиск новых надежных маркеров НИ ЦНС — важной научной целью.МАТЕРИАЛ И МЕТОДЫ Представленное исследование было проспективным и состояло из двух частей. Целью первой части было определить нормальный уровень пресепсина (ПСП) в спинномозговой жидкости (СМЖ). Для определения нормального уровня ПСП в СМЖ исследовались образцы ликвора, полученные при спинномозговой анестезии во время плановых урологических и общехирургических операций. Целью второй части было изучение динамики ПСП в СМЖ у различных групп нейрореанимационных пациентов в зависимости от наличия НИ ЦНС и системной инфекции. Вместе с ПСП в ликворе исследовали цитоз, уровень глюкозы, лактата, проводили его микробиологическое исследование и полимеразную цепную реакцию (ПЦР), когда это было возможно. Исследование крови включало в себя ее клинический анализ, определение содержания в ней С-реактивного белка (СРБ), прокальцитонина (PCT) и ПСП. Статистический анализ проводился с использованием IBM SPSS версии 23.0.РЕЗУЛЬТАТЫ В первой части исследования для получения нормального уровня ПСП в СМЖ были исследованы 15 проб СМЖ у пациентов урологического или общехирургического профиля без поражения нервной системы. Уровень ПСП в СМЖ составил 50–100 пг/мл. Эти значения были приняты в качестве референсных для уровня ПСП в СМЖ в норме. Во второй части исследования были проанализированы 63 пары проб ликвора и крови у 19 нейрореанимационных пациентов. Все пары были разделены на 4 группы в соответствии с наличием в момент забора ликвора и крови НИ ЦНС и системной инфекции. При наличии НИ ЦНС и системной инфекции (группа 1) уровень ПСП в СМЖ составил 731,1±389,7 пг/мл. При отсутствии НИ ЦНС и наличии системной инфекции (группа 2) уровень ПСП в СМЖ составил 614,9±315 пг/мл. При наличии НИ ЦНС и отсутствии системной инфекции (группа 3) уровень ПСП в СМЖ составил 547,8±264,3 пг/мл. При отсутствии НИ ЦНС и системной инфекции (группа 4) уровень ПСП в СМЖ составил 406±203,1 пг/мл. ROC-анализ показал, что уровень ПСП в СМЖ выше 537 пг/мл у нейрореанимационных пациентов без системной инфекции означает наличие НИ ЦНС с чувствительностью 68,8% и специфичностью 85,7%.ВЫВОДЫ В норме уровень пресепсина в ликворе составляет 50–100 пг/мл. Уровень пресепсина в ликворе выше 537 пг/мл у нейрореанимационного пациента без системной инфекции статистически значимо означает наличие у него НИ ЦНС. При диагностике НИ ЦНС определение уровня пресепсина в ликворе должно подлежать анализу вместе с традиционными маркерами инфекции ЦНС в качестве дополнительного маркера. Необходимо проведение дальнейших исследований

Определение валидности шкалы дыхательных нарушений у пациентов с острым поражением нервной системы

The aim of study: to investigate validity of respiratory insufficiency scale (RIS) in patients with acute lesions of nervous system.Material and methods. The prospective observational study included neurocritical care patients (n=179), admitted to the resuscitation and intensive care unit with independent breathing and RIS score 1 and higher. Patients were assessed according to RIS every 12 hours during the the period of RICU stay until the beginning of artificial lung ventilation or transfer to a specialized department. The RIS score did not influence the physician's decision upon intubation. The treatment was performed in accordance with national and international recommendations.Depending on the tracheal intubation and ALV, patients were divided into 3 groups. Group I (n=65): 0% tracheal intubation and ALV; Group II (n =54): 42,6% cases of intubation and ALV; Group III (n=60): 100% patients requiring intubation and ALV.The statistical analysis was performed using Shapiro—Wilk test, Mann-Whitney test, Kruskal—Wallis test, Chi-squared test. The ROC analysis was carried out to determine the sensitivity and specificity of the RIS scale.Results. Patients with RIS score 1 — 2 did not require intubation and ALV. Patients with RIS 5 or more required urgent intubation and ALV. In patients with RIS score 3—4 the need for intubation and ALV was unpredictable. If RIS score 4 was sustainig during several hours, or if increased from 3 to 4, a patient required intubation and initiation of ALV.Conclusion. RIS helps objectify indications for intubation and ALV in patients with acute neural lesions.Цель исследования. Исследовать валидность шкалы дыхательных нарушений (ШДН) у пациентов с острым повреждением нервной системы.Материал и методы. В проспективное обсервационное исследование включили нейрореанимационных пациентов (n=179), поступавших в отделение реанимации и интенсивной терапии (ОРИТ) c сохраненным самостоятельным дыханием с оценкой по ШДН, равной 1 и более балла. Пациентов оценивали по ШДН каждые 12 ч до момента интубации трахеи или до перевода из ОРИТ в профильное отделение. Оценка пациента по ШДН не влияла на принятие решения лечащим врачом об интубации трахеи. Ведение пациентов осуществляли в соответствии с национальными и международными рекомендациями. Пациентов разделили на 3 группы в зависимости от степени необходимости произвести интубацию трахеи и искусственную вентиляцию легких (ИВЛ). В группе I (n=65) — 0% интубации трахеи и ИВЛ; в группе II (n=54) — 42,6% интубации трахеи и ИВЛ; в группе III (n=60) — 100% интубации трахеи и ИВЛ. Статистический анализ проводили с применением методик Шапиро–Уилка, тестов Манна–Уитни, Крускала–Уоллиса, критерия согласия Пирсона Хи-квадрат (χ2). ROC-анализ был проведен для определения чувствительности и специфичности ШДН-шкалы.Результаты. При оценке по ШДН, равной 1–2 балла, пациенты не нуждаются в интубации трахеи и ИВЛ, при сумме 5 и более баллов необходима незамедлительная интубация трахеи и ИВЛ, при сумме 3– 4 балла необходимость в ИВЛ непредсказуема. При сохраняющейся в течение нескольких часов оценке по ШДН, равной 4 баллам, или при ее увеличении с 3 баллов до 4 необходимы интубация трахеи и начало ИВЛ.Заключение. У пациентов с острым поражением нервной системы применение оценки дыхательных нарушений по ШДН позволяет объективизировать показания для интубации трахеи и начала ИВЛ

Local Ca2+ Entry Via Orai1 Regulates Plasma Membrane Recruitment of TRPC1 and Controls Cytosolic Ca2+ Signals Required for Specific Cell Functions

Store-operated Ca2+ entry (SOCE) has been associated with two types of channels: CRAC channels that require Orai1 and STIM1 and SOC channels that involve TRPC1, Orai1, and STIM1. While TRPC1 significantly contributes to SOCE and SOC channel activity, abrogation of Orai1 function eliminates SOCE and activation of TRPC1. The critical role of Orai1 in activation of TRPC1-SOC channels following Ca2+ store depletion has not yet been established. Herein we report that TRPC1 and Orai1 are components of distinct channels. We show that TRPC1/Orai1/STIM1-dependent ISOC, activated in response to Ca2+ store depletion, is composed of TRPC1/STIM1-mediated non-selective cation current and Orai1/STIM1-mediated ICRAC; the latter is detected when TRPC1 function is suppressed by expression of shTRPC1 or a STIM1 mutant that lacks TRPC1 gating, STIM1(684EE685). In addition to gating TRPC1 and Orai1, STIM1 mediates the recruitment and association of the channels within ER/PM junctional domains, a critical step in TRPC1 activation. Importantly, we show that Ca2+ entry via Orai1 triggers plasma membrane insertion of TRPC1, which is prevented by blocking SOCE with 1 µM Gd3+, removal of extracellular Ca2+, knockdown of Orai1, or expression of dominant negative mutant Orai1 lacking a functional pore, Orai1-E106Q. In cells expressing another pore mutant of Orai1, Orai1-E106D, TRPC1 trafficking is supported in Ca2+-containing, but not Ca2+-free, medium. Consistent with this, ICRAC is activated in cells pretreated with thapsigargin in Ca2+-free medium while ISOC is activated in cells pretreated in Ca2+-containing medium. Significantly, TRPC1 function is required for sustained KCa activity and contributes to NFκB activation while Orai1 is sufficient for NFAT activation. Together, these findings reveal an as-yet unidentified function for Orai1 that explains the critical requirement of the channel in the activation of TRPC1 following Ca2+ store depletion. We suggest that coordinated regulation of the surface expression of TRPC1 by Orai1 and gating by STIM1 provides a mechanism for rapidly modulating and maintaining SOCE-generated Ca2+ signals. By recruiting ion channels and other signaling pathways, Orai1 and STIM1 concertedly impact a variety of critical cell functions that are initiated by SOCE

A High-Level Language for Rule-Based Modelling

Rule-based languages such as Kappa excel in their support for handling the combinatorial complexities prevalent in many biological systems, including signalling pathways. But Kappa provides little structure for organising rules, and large models can therefore be hard to read and maintain. This paper introduces a high-level, modular extension of Kappa called LBS-κ. We demonstrate the constructs of the language through examples and three case studies: a chemotaxis switch ring, a MAPK cascade, and an insulin signalling pathway. We then provide a formal definition of LBS-κ through an abstract syntax and a translation to plain Kappa. The translation is implemented in a compiler tool which is available as a web application. We finally demonstrate how to increase the expressivity of LBS-κ through embedded scripts in a general-purpose programming language, a technique which we view as generally applicable to other domain specific languages

Efficient and Correct Stencil Computation via Pattern Matching and Static Typing

Stencil computations, involving operations over the elements of an array, are a common programming pattern in scientific computing, games, and image processing. As a programming pattern, stencil computations are highly regular and amenable to optimisation and parallelisation. However, general-purpose languages obscure this regular pattern from the compiler, and even the programmer, preventing optimisation and obfuscating (in)correctness. This paper furthers our work on the Ypnos domain-specific language for stencil computations embedded in Haskell. Ypnos allows declarative, abstract specification of stencil computations, exposing the structure of a problem to the compiler and to the programmer via specialised syntax. In this paper we show the decidable safety guarantee that well-formed, well-typed Ypnos programs cannot index outside of array boundaries. Thus indexing in Ypnos is safe and run-time bounds checking can be eliminated. Program information is encoded as types, using the advanced type-system features of the Glasgow Haskell Compiler, with the safe-indexing invariant enforced at compile time via type checking