Methodological approaches for minimization laboratory researches in diagnostics of platelets hyperaggregation

The way of diagnostics of hyperactivity platelets, not demanding special conditions of haemocoagulation shifts in a vascular wall in vitro is offered. The way is based on biochemical research of plasma of blood of the patient used for routine haemostaziological of tests; with the subsequent calculation of a diagnostic index by solving rule. It is surveyed 65 patients in total. The correlation analysis of a degree of aggregation platelets and pH-plasmas of blood at women is carried out The method of the discriminantal analysis deduces solving rule of conformity of values pH-plasmas and hyperaggregations of platelets.Sol\ring rule: Д =constl X PPP-const2.Where: Д — diagnostic significant index; constl and const2-constants; PPP - value of a parameter pH poor platelets plasmas of the researched patient. The way does not demand the expensive material base, special reactants, is convenient in execution at use of a minimum quantity of a researched material, provides reception of the necessary information within one hour from the moment of a fence of blood of the patient, we reproduce in laboratory of any level.Предложен способ диагностики гиперактивности тромбоцитов, не требующий специальных условий моделирования реологии в сосудистой стенке in vitro. Способ основан на биохимическом исследовании плазмы крови пациента, используемой для рутинных гемостазиологических тестов, с последующим подсчетом диагностического индекса по решающему правилу. Всего обследовано 65 пациентов. Проведен корреляционный анализ степени агрегации тромбоцитов и pH плазмы крови пациентов. Методом дискриминантного анализа выведено решающее правило соответствия значений pH плазмы и гиперагрегации тромбоцитов: Д = constlXPPP- const2 где: Д - диагностически значимый индекс; constl и const2- постоянные величины: РРР - значение показателя pH бедной тромбоцитами плазмы исследуемого пациента. Способ не требует дорогостоящей материально-технической базы, специальных реактивов, удобен в исполнении при использовании минимального количества исследуемого материала, обеспечивает получение необходимой информации в течение часа с момента забора крови пациента, воспроизводим в лаборатории любого уровня

SORPTION OF SB (III) FROM SOLUTIONS USING MODIFIED MONTMORILLONITE

The possibility of antimony (III) sorption on composite sorbents created by modification of montmorillonite was studied. Montmorillonite modified with Fe3O4 nanoparticles and cationic surfactant showed a better degree of antimony (III) sorption compared to other modifications

Study of the Sorption of Arsenic(III) by Montmorillonite

The possibility of arsenic(III) sorption on composite sorbents created by modification of montmorillonite was studied. Montmorillonite modified with Fe3O4 nanoparticles and cationic surfactant showed a better degree of arsenic sorption compared to other modifications.Работа выполнена по Государственному заданию ИМЕТ УрО РАН в рамках Программы фундаментальных исследований государственных академий

Widespread Expression of BORIS/CTCFL in Normal and Cancer Cells

BORIS (CTCFL) is the paralog of CTCF (CCCTC-binding factor; NM_006565), a ubiquitously expressed DNA-binding protein with diverse roles in gene expression and chromatin organisation. BORIS and CTCF have virtually identical zinc finger domains, yet display major differences in their respective C- and N-terminal regions. Unlike CTCF, BORIS expression has been reported only in the testis and certain malignancies, leading to its classification as a “cancer-testis” antigen. However, the expression pattern of BORIS is both a significant and unresolved question in the field of DNA binding proteins. Here, we identify BORIS in the cytoplasm and nucleus of a wide range of normal and cancer cells. We compare the localization of CTCF and BORIS in the nucleus and demonstrate enrichment of BORIS within the nucleolus, inside the nucleolin core structure and adjacent to fibrillarin in the dense fibrillar component. In contrast, CTCF is not enriched in the nucleolus. Live imaging of cells transiently transfected with GFP tagged BORIS confirmed the nucleolar accumulation of BORIS. While BORIS transcript levels are low compared to CTCF, its protein levels are readily detectable. These findings show that BORIS expression is more widespread than previously believed, and suggest a role for BORIS in nucleolar function

De novo CCND2 mutations leading to stabilization of cyclin D2 cause megalencephaly-polymicrogyria-polydactyly-hydrocephalus syndrome

Activating mutations in genes encoding phosphatidylinositol 3-kinase (PI3K)-AKT pathway components cause megalencephaly-polymicrogyria-polydactyly-hydrocephalus syndrome (MPPH, OMIM 603387). Here we report that individuals with MPPH lacking upstream PI3K-AKT pathway mutations carry de novo mutations in CCND2 (encoding cyclin D2) that are clustered around a residue that can be phosphorylated by glycogen synthase kinase 3β (GSK-3β). Mutant CCND2 was resistant to proteasomal degradation in vitro compared to wild-type CCND2. The PI3K-AKT pathway modulates GSK-3β activity, and cells from individuals with PIK3CA, PIK3R2 or AKT3 mutations showed similar CCND2 accumulation. CCND2 was expressed at higher levels in brains of mouse embryos expressing activated AKT3. In utero electroporation of mutant CCND2 into embryonic mouse brains produced more proliferating transfected progenitors and a smaller fraction of progenitors exiting the cell cycle compared to cells electroporated with wild-type CCND2. These observations suggest that cyclin D2 stabilization, caused by CCND2 mutation or PI3K-AKT activation, is a unifying mechanism in PI3K-AKT–related megalencephaly syndromes

Selective cancer-germline gene expression in pediatric brain tumors

Cancer-germline genes (CGGs) code for immunogenic antigens that are present in various human tumors and can be targeted by immunotherapy. Their expression has been studied in a wide range of human tumors in adults. We measured the expression of 12 CGGs in pediatric brain tumors, to identify targets for therapeutic cancer vaccines. Real Time PCR was used to quantify the expression of genes MAGE-A1, MAGE-A2, MAGE-A3, MAGE-A4, MAGE-A6, MAGE-A10, MAGE-A12, MAGE-C2, NY-ESO-1 and GAGE-1,2,8 in 50 pediatric brain tumors of different histological subtypes. Protein expression was examined with immunohistochemistry. Fifty-five percent of the medulloblastomas (n = 11), 86% of the ependymomas (n = 7), 40% of the choroid plexus tumors (n = 5) and 67% of astrocytic tumors (n = 27) expressed one or more CGGs. Immunohistochemical analysis confirmed qPCR results. With exception of a minority of tumors, the overall level of CGG expression in pediatric brain tumors was low. We observed a high expression of at least one CGG in 32% of the samples. CGG-encoded antigens are therefore suitable targets in a very selected group of pediatric patients with a brain tumor. Interestingly, glioblastomas from adult patients expressed CGGs more often and at significantly higher levels compared to pediatric glioblastomas. This observation is in line with the notion that pediatric and adult glioblastomas develop along different genetic pathways

A Cell Cycle Role for the Epigenetic Factor CTCF-L/BORIS

CTCF is a ubiquitous epigenetic regulator that has been proposed as a master keeper of chromatin organisation. CTCF-like, or BORIS, is thought to antagonise CTCF and has been found in normal testis, ovary and a large variety of tumour cells. The cellular function of BORIS remains intriguing although it might be involved in developmental reprogramming of gene expression patterns. We here unravel the expression of CTCF and BORIS proteins throughout human epidermis. While CTCF is widely distributed within the nucleus, BORIS is confined to the nucleolus and other euchromatin domains. Nascent RNA experiments in primary keratinocytes revealed that endogenous BORIS is present in active transcription sites. Interestingly, BORIS also localises to interphase centrosomes suggesting a role in the cell cycle. Blocking the cell cycle at S phase or mitosis, or causing DNA damage, produced a striking accumulation of BORIS. Consistently, ectopic expression of wild type or GFP- BORIS provoked a higher rate of S phase cells as well as genomic instability by mitosis failure. Furthermore, downregulation of endogenous BORIS by specific shRNAs inhibited both RNA transcription and cell cycle progression. The results altogether suggest a role for BORIS in coordinating S phase events with mitosis

Plexin-A2 and its ligand, Sema6A, control nucleus-centrosome coupling in migrating granule cells.

During their migration, cerebellar granule cells switch from a tangential to a radial mode of migration. We have previously demonstrated that this involves the transmembrane semaphorin Sema6A. We show here that plexin-A2 is the receptor that controls Sema6A function in migrating granule cells. In plexin-A2-deficient (Plxna2(-/-)) mice, which were generated by homologous recombination, many granule cells remained in the molecular layer, as we saw in Sema6a mutants. A similar phenotype was observed in mutant mice that were generated by mutagenesis with N-ethyl-N-nitrosourea and had a single amino-acid substitution in the semaphorin domain of plexin-A2. We found that this mutation abolished the ability of Sema6A to bind to plexin-A2. Mouse chimera studies further suggested that plexin-A2 acts in a cell-autonomous manner. We also provide genetic evidence for a ligand-receptor relationship between Sema6A and plexin-A2 in this system. Using time-lapse video microscopy, we found that centrosome-nucleus coupling and coordinated motility were strongly perturbed in Sema6a(-/-) and Plxna2(-/-) granule cells. This suggests that semaphorin-plexin signaling modulates cell migration by controlling centrosome positioning

Regulators of the Actin Cytoskeleton Mediate Lethality in a Caenorhabditis elegans dhc-1 Mutant

Both LIS-1 and the dynein heavy-chain DHC-1 are required for integrity of the actin cytoskeleton in Caenorhabditis elegans. An RNAi screen revealed that knockdown of other actin regulators, including actin-capping protein genes and prefoldin subunit genes, suppresses dhc-1(or195ts)–induced lethality