A major cellular substrate for protein kinases, annexin II, is a DNA-binding protein

AbstractWe have screened a human cDNA expression library in λgt11 for clones encoding Alu-binding proteins using direct binding of labeled Alu DNA to recombinant phage lysates fixed on a membrane, and isolated a clone 98% identical in sequence to the well-known substrate of protein kinases, annexin II, which was suggested earlier to play a role in transduction of mitogenic signals and DNA replication. A diagnostic property of annexins is their binding to phospholipids in the presence of calcium ions, and we have found that the interaction of proteins of human nuclear extracts with Alu subsequences is suppressed by Ca/phosphatidylserine liposomes, suggesting overlapping of Ca/phospholipid- and DNA-binding domains in annexin II

Individual differences in toddlers' social understanding and prosocial behavior: Disposition or socialization?

We examined how individual differences in social understanding contribute to variability in early-appearing prosocial behavior. Moreover, potential sources of variability in social understanding were explored and examined as additional possible predictors of prosocial behavior. Using a multi-method approach with both observed and parent-report measures, 325 children aged 18-30 months were administered measures of social understanding (e.g., use of emotion words; self-understanding), prosocial behavior (in separate tasks measuring instrumental helping, empathic helping, and sharing, as well as parent-reported prosociality at home), temperament (fearfulness, shyness, and social fear), and parental socialization of prosocial behavior in the family. Individual differences in social understanding predicted variability in empathic helping and parent-reported prosociality, but not instrumental helping or sharing. Parental socialization of prosocial behavior was positively associated with toddlers' social understanding, prosocial behavior at home, and instrumental helping in the lab, and negatively associated with sharing (possibly reflecting parents' increased efforts to encourage children who were less likely to share). Further, socialization moderated the association between social understanding and prosocial behavior, such that social understanding was less predictive of prosocial behavior among children whose parents took a more active role in socializing their prosociality. None of the dimensions of temperament was associated with either social understanding or prosocial behavior. Parental socialization of prosocial behavior is thus an important source of variability in children's early prosociality, acting in concert with early differences in social understanding, with different patterns of influence for different subtypes of prosocial behavior

EXAMINING LOCALLY EXPRESSED mRNA OF INFLAMMATORY MEDIATOR GENES IN A MODEL OF RETINAL PIGMENT EPITHELIUM ATROPHY AND RETINAL DEGENERATION INDUCED BY SUBRETINAL SALINE INJECTION IN RABBITS

Degenerative-dystrophic retinal diseases, particularly age-related macular degeneration (AMD), are now considered to be the lead cause of blindness and low vision in developed countries, with a steadily increasing trend. Recent publications provide evidence for the involvement of inflammatory mechanisms in TMD development and progression unveiled due to advances in innate and adaptive immunity research. However, the immunopathogenesis of atrophic AMD form, “geographic atrophy” (GA) remains largely unstudied. Objective: to investigate local mRNA expression of inflammatory cytokines IL-1β, IL-18, CCL2/MCP-1 in a model of RPE atrophy induced after subretinal injection of 0.9% sodium chloride solution in experimental rabbits. The investigation was carried out in tissue complex retina-RPE-choroid (TC) samples isolated from eyes of 23 albino New Zealand rabbits after modeling RPE atrophy by subretinal injection of 0.9% sodium chloride solution and 5 healthy rabbits lacking eye lesions. Animals in the experimental group (one week before surgical intervention, in the early period, and in the period of sustained RPE atrophy formation) and controls were subjected to optical coherence tomography (OCT) and ocular fundus autofluorescence (FAF). Evaluation of proinflammatory cytokine gene expression levels in TC was performed by RT-PCR. Results. Subretinal injection of 0.01 ml of 0.9% sodium chloride solution induced experimental RPE atrophy development in rabbits vs. control that was associated with multidirectional changes of IL-1β, IL-18, MCP-1/CCL2 gene mRNA expression. Three types of response in the TC, formed during development of atrophic changes and determined by the value of local cytokine gene expression were characterized: 1) hypo/ no response – decreased/no expression; 2) normal response – moderate increase; 3) hyper response – overexpression. 69.6% of animals with persistent atrophy had a moderate to hypertrophic increase in locally expressed mRNA MCP-1/CCL2, whereas 30% cases had significantly increased IL-1β mRNA expression – factors damaging the blood-retinal barrier and contributing to posterior segment immune privilege. It should be taken into account while developing new strategies for treatment of ophthalmic pathology, in particular the currently actively studied and tested options for RPE stem cell transplantation into subretinal space. The data obtained may be useful to investigate various types of RPE atrophy and develop new strategies of ophthalmopathology treatment in preclinical studies

Fatty Acids of Erythrocyte Membranes and Blood Serum in Differential Diagnosis of Inflammatory Bowel Diseases

Aim: to study fatty acid levels in erythrocyte membranes (RBC) and blood serum (BS) in patients with inflammatory bowel diseases (IBDs) to develop differential diagnostic models including fatty acids as biomarkers to distinguish between nosological entities of IBDs (ulcerative colitis — UC, Crohn's disease — CD, unclassified colitis — UCC).Materials and methods. We examined 110 patients (mean age 37,7 ± 12,1 years) with IBDs and 53 healthy patients in control group (43,3 ± 11,7 years). The IBDs group included 50 patients with UC, 41 patients with CD, 19 patients with UCC. An exacerbation of the disease was revealed in 42 patients (84 %) with UC, 34 patients with CD (82.9 %) and 11 people with UCC (57.9 %). The study of fatty acids (FA) composition of RBC membranes and BS was carried out using GC/MS system based on three Agilent 7000B quadrupoles (USA).Results. The most significant for distinguishing active UC from CD exacerbation were serum levels of elaidin (p = 0.0006); docosatetraenoic (n-6) (p = 0.004); docodienic (n-6) (p = 0.009); omega-3/omega-6 ratio (p = 0.02); docosapentaenoic (n-3) (p = 0.03); the sum of eicosapentaenoic and docosahexaenoic (p = 0.03), as well as the content of RBC lauric FA (p = 0.04) (AUC — 0.89, sensitivity — 0.91, specificity — 0.89, diagnostic accuracy — 0.91). To distinguish active UC from the same of UCC, the following serum FA were found to be significant: alpha-linolenic; saturated (pentadecanoic, palmitic, stearic, arachidic); monounsaturated (palmitoleic, oleic); omega-6 (hexadecadienic, arachidonic) (p = 0.00000011—0.03300000) (AUC — 0.995, sensitivity — 0.98, specificity — 0.96, diagnostic accuracy — 0.97). The most significant in distinguishing patients with active CD from UCC exacerbation were levels of the following FA: alpha-linolenic; palmitoleic; oleic; the amount of saturated fatty acids (SFA); total unsaturated fatty acids (UFA); stearic; monounsaturated fatty acids (MUFA) amount; SFA/UFA; SFA/PUFA (polyunsaturated fatty acids); linoleic; total PUFA n6; lauric; arachidic acid (p = 0.0000000017–0.030000000) (AUC — 0.914, sensitivity — 0.90, specificity — 0.87, diagnostic accuracy — 0.91).Conclusion. The study of FA levels in groups with different nosological forms of IBDs using complex statistical analysis, including machine learning methods, made it possible to create diagnostic models that differentiate CD, UC and UCC in the acute stage with high accuracy. The proposed approach is promising for the purposes of differential diagnosis of nosological forms of IBDs

Synovial changes detected by ultrasound in people with knee osteoarthritis - a meta-analysis of observational studies

Objectives To examine the prevalence of synovial effusion, synovial hypertrophy and positive Doppler signal (DS) detected by ultrasound (US) in people with knee osteoarthritis (OA) and/or knee pain compared to that in the general population. Method A systematic literature search was undertaken in Medline, EMBASE, Allied and Complementary Medicine, PubMed Web of Science, and SCOPUS databases in May 2015. Frequencies of US abnormalities in people with knee OA/pain, in the general population or asymptomatic controls were pooled using the random effects model. Publication bias and heterogeneity between studies were examined. Results Twenty four studies in people with knee pain/OA and five studies of the general population or asymptomatic controls met the inclusion criteria. The pooled prevalence of US effusion, synovial hypertrophy and positive DS in people with knee OA/pain were 51.5% (95% CI 40.2 to 62.8), 41.5% (26.3–57.5) and 32.7% (8.34–63.24), respectively, which were higher than those in the general population or asymptomatic controls (19.9% (95%CI 7.8–35.3%), 14.5% (0–58.81), and 15.8 (3.08–35.36), respectively). People with knee OA (ACR criteria or radiographic OA) had greater prevalence of US abnormalities than people with knee pain (P = 0.037, P = 0.010 and P = 0.009, respectively). Conclusions US detected effusion, synovial hypertrophy and DS are more common in people with knee OA/pain, compared to the general population. These abnormalities relate more to presence of OA structural changes than to pain

Об адекватных зонах истинного ВГД в здоровых и глаукомных глазах

PURPOSE. The study was conducted to reveal using the Ocular Response Analyzer (ORA) the possible ranges of mean values of the “rigidity” and “fluctuation” criteria for fibrous membrane of healthy and glaucoma eyes with consideration of the age periods according to the classification by WHO, and to identify adequate true intraocular pressure (IOP) zones corresponding to these ranges.METHODS. The study consisted of a theoretical analysis of clinical measurements of rigidity and fluctuation of the fibrous membrane, the current level of true IOP and the individual level of IOP calculated in youth using ORA by the method of dynamic diagnosis of Koshitsa-Svetlova, and involved in total 674 healthy and 518 glaucoma eyes from individuals aged 18 to 90 years, who were distributed by age periods according to WHO.RESULTS. A "step pattern" of the distribution of Average values of rigidity and fluctuation in healthy and glaucoma eyes were distributed in a “step pattern”, which made it possible to rank the IOP zones adequately to these steps, taking into account the age periods according to WHO. The following adequate ranges of IOP levels for healthy and glaucoma eyes were identified: low IOP zone (up to 13 mm Hg); medium IOP zone (14–20 mm Hg); elevated IOP zone (21–26 mm Hg); high IOP zone (27–32 mm Hg); IOP subcompensation zone (33–39 mm Hg) and uncompensated IOP zone (≥40 mm Hg). The "steps" of the average values of rigidity, fluctuations, and the IOP ranges adequate to them do not intersect. The current value of the rigidity of the fibrous membrane and the IOP value calculated in youth make it possible to reliably attribute each healthy or glaucoma eye to its individual IOP zone.CONCLUSION. Rigidity of the fibrous membrane consistently determines the level of IOP (p>0.001), while the rigidity and fluctuation of the sclera directly influence its current level. The fundamental criteria — rigidity and fluctuation of the fibrous membrane of the eye — do not depend on the central corneal thickness and objectively determine the current functional state of the fibrous membrane. The ability to objectively and reliably determine whether a healthy or glaucoma eye belongs to its individual IOP zone is particularly important for the polyclinic network. The time such express diagnostics takes is 0.02 seconds.ЦЕЛЬ. С помощью пневмоанализатора Ocular Response Analyzer (ORA) выявить у фиброзной оболочки глаза (ФОГ) возможные диапазоны средних значений функциональных критериев «ригидность» и «флуктуация» в здоровых и глаукомных глазах с учетом возрастных периодов старения по классификации ВОЗ и определить соответствующие этим диапазонам адекватные диапазоны истинного внутриглазного давления (ВГД).МЕТОДЫ. Теоретический анализ результатов измерения в клинике значений ригидности и флуктуации ФОГ, текущего уровня истинного ВГД и расчетного индивидуального уровня ВГД у пациента в молодости, полученных с помощью ORA по способу динамической диагностики Кошица-Светловой у 674 здоровых и 518 глаукомных глаз в возрасте 18–90 лет, распределенных согласно периодам старения по ВОЗ.РЕЗУЛЬТАТЫ. Выявлена «ступенчатая закономерность» распределения средних значений ригидности и флуктуации в здоровых и глаукомных глазах, что позволило ранжировать зоны ВГД адекватно этим ступеням с учетом возрастных периодов старения по ВОЗ. Объективно выявлены следующие адекватные диапазоны уровня ВГД для здоровых и глаукомных глаз: зона низкого ВГД (до 13 мм рт.ст.); зона среднего ВГД (14–20 мм рт.ст.); зона повышенного ВГД (21–26 мм рт.ст); зона высокого ВГД (27–32 мм рт.ст.); зона субкомпенсации ВГД (33–39 мм рт.ст.) и зона некомпенсации ВГД (≥40 мм рт.ст.). «Ступени» средних значений ригидности, флуктуации и адекватные им диапазоны уровня ВГД не пересекаются. Текущее значение ригидности ФОГ и расчетное значение уровня ВГД в молодости позволяют достоверно отнести каждый здоровый или глаукомный глаз к его индивидуальной зоне ВГД.ЗАКЛЮЧЕНИЕ. Значение ригидности ФОГ закономерно определяет уровень ВГД (р<0,001), а ригидность и флуктуация склеры напрямую формируют его текущий уровень. Фундаментальные критерии ригидности и флуктуации ФОГ не зависят от центральной толщины роговицы и объективно определяют текущее функциональное состояние ФОГ. Возможность объективно и достоверно определять принадлежность здорового или глаукомного глаза к его индивидуальной зоне ВГД представляется особенно важной для поликлинической сети. Время проведения такой экспресс-диагностики составляет 0,02 секунды

Intestinal epithelial stem cells do not protect their genome by asymmetric chromosome segregation

The idea that stem cells of adult tissues with high turnover are protected from DNA replication-induced mutations by maintaining the same 'immortal' template DNA strands together through successive divisions has been tested in several tissues. In the epithelium of the small intestine, the provided evidence was based on the assumption that stem cells are located above Paneth cells. The results of genetic lineage-tracing experiments point instead to crypt base columnar cells intercalated between Paneth cells as bona fide stem cells. Here we show that these cells segregate most, if not all, of their chromosomes randomly, both in the intact and in the regenerating epithelium. Therefore, the 'immortal' template DNA strand hypothesis does not apply to intestinal epithelial stem cells, which must rely on other strategies to avoid accumulating mutations

H2AX phosphorylation screen of cells from radiosensitive cancer patients reveals a novel DNA double-strand break repair cellular phenotype

BACKGROUND: About 1-5% of cancer patients suffer from significant normal tissue reactions as a result of radiotherapy (RT). It is not possible at this time to predict how most patients' normal tissues will respond to RT. DNA repair dysfunction is implicated in sensitivity to RT particularly in genes that mediate the repair of DNA double-strand breaks (DSBs). Phosphorylation of histone H2AX (phosphorylated molecules are known as gammaH2AX) occurs rapidly in response to DNA DSBs, and, among its other roles, contributes to repair protein recruitment to these damaged sites. Mammalian cell lines have also been crucial in facilitating the successful cloning of many DNA DSB repair genes; yet, very few mutant cell lines exist for non-syndromic clinical radiosensitivity (RS).\ud \ud METHODS: Here, we survey DNA DSB induction and repair in whole cells from RS patients, as revealed by gammaH2AX foci assays, as potential predictive markers of clinical radiation response.\ud \ud RESULTS: With one exception, both DNA focus induction and repair in cell lines from RS patients were comparable with controls. Using gammaH2AX foci assays, we identified a RS cancer patient cell line with a novel ionising radiation-induced DNA DSB repair defect; these data were confirmed by an independent DNA DSB repair assay.\ud \ud CONCLUSION: gammaH2AX focus measurement has limited scope as a pre-RT predictive assay in lymphoblast cell lines from RT patients; however, the assay can successfully identify novel DNA DSB repair-defective patient cell lines, thus potentially facilitating the discovery of novel constitutional contributions to clinical RS

A multiplatform approach identifies miR-152-3p as a common epigenetically regulated onco-suppressor in prostate cancer targeting TMEM97

Prostate cancer (PCa) is a major cause of morbidity and mortality in men worldwide. MicroRNAs are globally downregulated in PCa, especially in poorly differentiated tumors. Nonetheless, the underlying mechanisms are still elusive. Herein, using combined analysis of microRNAs expression and genomewide DNA methylation, we aimed to identify epigenetically downregulated microRNAs in PCa.Research Center of Portuguese Oncology Institute of Porto (FB-GEBC-27 and 19-CI-IPOP-2016). JR-C and CSG are supported by FCT- Fundação para a Ciência e Tecnologia PhD fellowships (SFRH/BD/71293/2010 and SFRH/BD/92786/2013), SS is supported by a PhD fellowship IPO/ESTIMA-1 NORTE-01-0145-FEDER-000027, and IG is a research fellow from the strategic funding of FCT (PCT: PEst- UID/DTP/00776/2013 and COMPETE: POCI-01-0145-FEDER-006868). BMC is funded by FCT-Fundação para a Ciência e a Tecnologia (IF/00601/2012)info:eu-repo/semantics/publishedVersio