ОЦІНКА ОКИСНОГО СТРЕСУ І РІВЕНЬ МОЛЕКУЛЯРНИХ УШКОДЖЕНЬ ПРИ ПОЗАЛІКАРНЯНІЙ ПНЕВМОНІЇ В ДІТЕЙ ІЗ ЙОДОДЕФIЦИТНОГО РЕГІОНУ

The aim of the study ‒ to learn the state of antioxidant protection and the subcellular distribution of iodine in children with moderate and severe course of community-acquired pneumonia.Materials and Methods. Seventy children with community-acquired pneumonia were examined. All children were examined subcellular iodine in the blood, the state of the antioxidant defense system, the state of stress-responsive systems and the level of molecular damage were assessed.Results and Discussion. A lower concentration of organified iodine was found (by 12 % and 59 %, respectively) and a high concentration of inorganic iodine (21 % and 48 %, respectively) in moderate and severe course pneumonia in compare to the control group. The severe course of pneumonia was accompanied by activation of catalase, suppression of superoxide dismutase, and against the background of increased oxyradical content, intensive formation of oxidative modification of proteins, and enhancement of DNA fragmentation. The revealed changes tend to decrease depending on the severity of pneumonia.Conclusions. The distribution of intracellular pool of iodine in the body are directly proportional to the severity of communityacquired pneumonia. The heavy flow of pneumonia contributes to a sharp increase in the concentration of inorganic iodine with a simultaneous decrease in the level of organic iodine in the blood. Fluctuations in the indices of oxidative stress and the level of molecular damage in children with community-acquired pneumonia correlate with the severity of the disease.Цель исследования ‒ изучить состояние антиоксидантной защиты при внебольничной пневмонии средней степени тяжести и тяжелом течении этого заболевания во взаимосвязи с особенностями субклеточного распределения йода в организме ребенка.Материалы и методы. Обследовано 70 детей с внегоспитальной пневмонией. Исследовали йодную обеспеченность пациентов путем изучения субклеточного распределения йода в крови, оценивали состояние системы антиоксидантной защиты, состояние стрессзависимых систем и уровень молекулярных повреждений.Результаты исследования и их обсуждение. Установлено меньшее содержание органифицированного йода (на 12 и 59 % соответственно) и высокую концентрацию в крови неорганического йода (21 и 48 % соответственно) при пневмонии средней степени тяжести и тяжелом течении этого заболевания относительно группы контроля. Тяжелое течение пневмонии сопровождалось активацией каталазы, угнетением супероксиддисмутазы на фоне увеличения содержания оксирадикалов, интенсивного образования окислительномодифицированных белков и усиления фрагментации ДНК. Выявленные изменения имеют тенденцию к уменьшению в зависимости от степени тяжести пневмонии.Выводы. Выявленные изменения показателей внутриклеточного пула йода и его распределение в организме прямо пропорциональны тяжести внебольничной пневмонии. Тяжелое течение пневмонии способствует резкому повышению концентрации неорганического йода с одновременным снижением уровня органического йода в крови. Колебания показателей окислительного стресса и уровень молекулярных повреждений у детей с внегоспитальной пневмонией коррелируют с тяжестью заболевания.Мета дослідження – вивчити стан антиоксидантного захисту при позалікарняній пневмонії середнього ступеня тяжкості й тяжкому перебігу цього захворювання у взаємозв’язку з особливостями субклітинного розподілу йоду в організмі дитини.Матеріали та методи. Обстежено 70 дітей з позалікарняною пневмонією. Досліджували йодну забезпеченість пацієнтів шляхом вивчення субклітинного розподілу йоду в крові, оцінювали стан системи антиоксидантного захисту, стан стресзалежних систем і рівень молекулярних ушкоджень.Результати дослідження та їх обговорення. Встановлено менший вміст органіфікованого йоду (на 12 і 59 % відповідно) та високу концентрацію в крові неорганічного йоду (21 і 48 % відповідно) при пневмонії середнього ступеня тяжкості й тяжкому перебігу цього захворювання щодо групи контролю. Тяжкий перебіг пневмонії супроводжувався активацією каталази, пригніченням супероксиддисмутази на тлі збільшення вмісту оксирадикалів, інтенсивного утворення окисномодифікованих білків і посилення фрагментації ДНК. Виявлені зміни мають тенденцію до зменшення залежно від ступеня тяжкості пневмонії.Висновки. Виявлені зміни показників внутрішньоклітинного пулу йоду і його розподіл в організмі прямо пропорційні тяжкості позалікарняної пневмонії. Тяжкий перебіг пневмонії сприяє різкому підвищенню концентрації неорганічного йоду з одночасним зниженням рівня органічного йоду в крові. Коливання показників окисного стресу і рівень молекулярних ушкоджень у дітей з позалікарняною пневмонією корелюють із тяжкістю захворювання

Spina bifida-predisposing heterozygous mutations in Planar Cell Polarity genes and Zic2 reduce bone mass in young mice

Fractures are a common comorbidity in children with the neural tube defect (NTD) spina bifida. Mutations in the Wnt/planar cell polarity (PCP) pathway contribute to NTDs in humans and mice, but whether this pathway independently determines bone mass is poorly understood. Here, we first confirmed that core Wnt/PCP components are expressed in osteoblasts and osteoclasts in vitro. In vivo, we performed detailed µCT comparisons of bone structure in tibiae from young male mice heterozygous for NTD-associated mutations versus WT littermates. PCP signalling disruption caused by Vangl2 (Vangl2Lp/+) or Celsr1 (Celsr1Crsh/+) mutations significantly reduced trabecular bone mass and distal tibial cortical thickness. NTD-associated mutations in non-PCP transcription factors were also investigated. Pax3 mutation (Pax3Sp2H/+) had minimal effects on bone mass. Zic2 mutation (Zic2Ku/+) significantly altered the position of the tibia/fibula junction and diminished cortical bone in the proximal tibia. Beyond these genes, we bioinformatically documented the known extent of shared genetic networks between NTDs and bone properties. 46 genes involved in neural tube closure are annotated with bone-related ontologies. These findings document shared genetic networks between spina bifida risk and bone structure, including PCP components and Zic2. Genetic variants which predispose to spina bifida may therefore independently diminish bone mass

Effectiveness of Progressive and Resisted and Non-Progressive or Non-Resisted Exercise in Rotator Cuff Related Shoulder Pain: A Systematic Review and Meta-analysis of Randomised Controlled Trials

Objective: Synthesize evidence regarding effectiveness of progressive and resisted or non-progressive and non-resisted exercise compared with placebo or no treatment, in rotator cuff related pain. Data sources: English articles, searched in Cochrane CENTRAL, MEDLINE, EMBASE and CINAHL databases up until May 19, 2020. Methods: Randomized controlled trials in people with rotator cuff related pain comparing either progressive and resisted exercise or non-progressive and non-resisted exercise, with placebo or no treatment were included. Data extracted independently by two authors. Risk of bias appraised with the Cochrane Collaboration tool. Results: Seven trials (468 participants) were included, four trials (271 participants) included progressive and resisted exercise and three trials (197 participants) included non-progressive or non-resisted exercise. There was uncertain clinical benefit for composite pain and function (15 point difference, 95% CI 9 to 21, 100-point scale) and pain outcomes at >6weeks to 6months with progressive and resisted exercise compared to placebo or no treatment (comparison 1). For non-progressive or non-resisted exercise there was no significant benefit for composite pain and function (4 point difference, 95% CI –2 to 9, 100-point scale) and pain outcomes at >6weeks to 6months compared to placebo or no treatment (comparison 2). Adverse events were seldom reported and mild. Conclusions: There is uncertain clinical benefit for all outcomes with progressive and resisted exercise and no significant benefit with non-progressive and non-resisted exercise, versus no treatment or placebo at >6weeks to 6months. Findings are low certainty and should be interpreted with cautio

Wolbachia Bacteria Reside in Host Golgi-Related Vesicles Whose Position Is Regulated by Polarity Proteins

Wolbachia pipientis are intracellular symbiotic bacteria extremely common in various organisms including Drosophila melanogaster, and are known for their ability to induce changes in host reproduction. These bacteria are present in astral microtubule-associated vesicular structures in host cytoplasm, but little is known about the identity of these vesicles. We report here that Wolbachia are restricted only to a group of Golgi-related vesicles concentrated near the site of membrane biogenesis and minus-ends of microtubules. The Wolbachia vesicles were significantly mislocalized in mutant embryos defective in cell/planar polarity genes suggesting that cell/tissue polarity genes are required for apical localization of these Golgi-related vesicles. Furthermore, two of the polarity proteins, Van Gogh/Strabismus and Scribble, appeared to be present in these Golgi-related vesicles. Thus, establishment of polarity may be closely linked to the precise insertion of Golgi vesicles into the new membrane addition site

Genomic and Transcriptional Co-Localization of Protein-Coding and Long Non-Coding RNA Pairs in the Developing Brain

Besides protein-coding mRNAs, eukaryotic transcriptomes include many long non-protein-coding RNAs (ncRNAs) of unknown function that are transcribed away from protein-coding loci. Here, we have identified 659 intergenic long ncRNAs whose genomic sequences individually exhibit evolutionary constraint, a hallmark of functionality. Of this set, those expressed in the brain are more frequently conserved and are significantly enriched with predicted RNA secondary structures. Furthermore, brain-expressed long ncRNAs are preferentially located adjacent to protein-coding genes that are (1) also expressed in the brain and (2) involved in transcriptional regulation or in nervous system development. This led us to the hypothesis that spatiotemporal co-expression of ncRNAs and nearby protein-coding genes represents a general phenomenon, a prediction that was confirmed subsequently by in situ hybridisation in developing and adult mouse brain. We provide the full set of constrained long ncRNAs as an important experimental resource and present, for the first time, substantive and predictive criteria for prioritising long ncRNA and mRNA transcript pairs when investigating their biological functions and contributions to development and disease

Wnt5a Regulates Ventral Midbrain Morphogenesis and the Development of A9–A10 Dopaminergic Cells In Vivo

Wnt5a is a morphogen that activates the Wnt/planar cell polarity (PCP) pathway and serves multiple functions during development. PCP signaling controls the orientation of cells within an epithelial plane as well as convergent extension (CE) movements. Wnt5a was previously reported to promote differentiation of A9–10 dopaminergic (DA) precursors in vitro. However, the signaling mechanism in DA cells and the function of Wnt5a during midbrain development in vivo remains unclear. We hereby report that Wnt5a activated the GTPase Rac1 in DA cells and that Rac1 inhibitors blocked the Wnt5a-induced DA neuron differentiation of ventral midbrain (VM) precursor cultures, linking Wnt5a-induced differentiation with a known effector of Wnt/PCP signaling. In vivo, Wnt5a was expressed throughout the VM at embryonic day (E)9.5, and was restricted to the VM floor and basal plate by E11.5–E13.5. Analysis of Wnt5a−/− mice revealed a transient increase in progenitor proliferation at E11.5, and a precociously induced NR4A2+ (Nurr1) precursor pool at E12.5. The excess NR4A2+ precursors remained undifferentiated until E14.5, when a transient 25% increase in DA neurons was detected. Wnt5a−/− mice also displayed a defect in (mid)brain morphogenesis, including an impairment in midbrain elongation and a rounded ventricular cavity. Interestingly, these alterations affected mostly cells in the DA lineage. The ventral Sonic hedgehog-expressing domain was broadened and flattened, a typical CE phenotype, and the domains occupied by Ngn2+ DA progenitors, NR4A2+ DA precursors and TH+ DA neurons were rostrocaudally reduced and laterally expanded. In summary, we hereby describe a Wnt5a regulation of Wnt/PCP signaling in the DA lineage and provide evidence for multiple functions of Wnt5a in the VM in vivo, including the regulation of VM morphogenesis, DA progenitor cell division, and differentiation of NR4A2+ DA precursors

Murine Dishevelled 3 Functions in Redundant Pathways with Dishevelled 1 and 2 in Normal Cardiac Outflow Tract, Cochlea, and Neural Tube Development

Dishevelled (Dvl) proteins are important signaling components of both the canonical β-catenin/Wnt pathway, which controls cell proliferation and patterning, and the planar cell polarity (PCP) pathway, which coordinates cell polarity within a sheet of cells and also directs convergent extension cell (CE) movements that produce narrowing and elongation of the tissue. Three mammalian Dvl genes have been identified and the developmental roles of Dvl1 and Dvl2 were previously determined. Here, we identify the functions of Dvl3 in development and provide evidence of functional redundancy among the three murine Dvls. Dvl3−/− mice died perinatally with cardiac outflow tract abnormalities, including double outlet right ventricle and persistent truncus arteriosis. These mutants also displayed a misorientated stereocilia in the organ of Corti, a phenotype that was enhanced with the additional loss of a single allele of the PCP component Vangl2/Ltap (LtapLp/+). Although neurulation appeared normal in both Dvl3−/− and LtapLp/+ mutants, Dvl3+/−;LtapLp/+ combined mutants displayed incomplete neural tube closure. Importantly, we show that many of the roles of Dvl3 are also shared by Dvl1 and Dvl2. More severe phenotypes were observed in Dvl3 mutants with the deficiency of another Dvl, and increasing Dvl dosage genetically with Dvl transgenes demonstrated the ability of Dvls to compensate for each other to enable normal development. Interestingly, global canonical Wnt signaling appeared largely unaffected in the double Dvl mutants, suggesting that low Dvl levels are sufficient for functional canonical Wnt signals. In summary, we demonstrate that Dvl3 is required for cardiac outflow tract development and describe its importance in the PCP pathway during neurulation and cochlea development. Finally, we establish several developmental processes in which the three Dvls are functionally redundant

Large-Scale Clonal Analysis Reveals Unexpected Complexity in Surface Ectoderm Morphogenesis

Background: Understanding the series of morphogenetic processes that underlie the making of embryo structures is a highly topical issue in developmental biology, essential for interpreting the massive molecular data currently available. In mouse embryo, long-term in vivo analysis of cell behaviours and movements is difficult because of the development in utero and the impossibility of long-term culture. Methodology/Principal Findings: We improved and combined two genetic methods of clonal analysis that together make practicable large-scale production of labelled clones. Using these methods we performed a clonal analysis of surface ectoderm (SE), a poorly understood structure, for a period that includes gastrulation and the establishment of the body plan. We show that SE formation starts with the definition at early gastrulation of a pool of founder cells that is already dorso-ventrally organized. This pool is then regionalized antero-posteriorly into three pools giving rise to head, trunk and tail. Each pool uses its own combination of cell rearrangements and mode of proliferation for elongation, despite a common clonal strategy that consists in disposing along the antero-posterior axis precursors of dorso-ventrally-oriented stripes of cells. Conclusions/Significance: We propose that these series of morphogenetic processes are organized temporally and spatially in a posterior zone of the embryo crucial for elongation. The variety of cell behaviours used by SE precursor cells indicates that these precursors are not equivalent, regardless of a common clonal origin and a common clonal strategy. Anothe

C. elegans VANG-1 Modulates Life Span via Insulin/IGF-1-Like Signaling

The planar cell polarity (PCP) pathway is highly conserved from Drosophila to humans and a PCP-like pathway has recently been described in the nematode Caenorhabditis elegans. The developmental function of this pathway is to coordinate the orientation of cells or structures within the plane of an epithelium or to organize cell-cell intercalation required for correct morphogenesis. Here, we describe a novel role of VANG-1, the only C. elegans ortholog of the conserved PCP component Strabismus/Van Gogh. We show that two alleles of vang-1 and depletion of the protein by RNAi cause an increase of mean life span up to 40%. Consistent with the longevity phenotype vang-1 animals also show enhanced resistance to thermal- and oxidative stress and decreased lipofuscin accumulation. In addition, vang-1 mutants show defects like reduced brood size, decreased ovulation rate and prolonged reproductive span, which are also related to gerontogenes. The germline, but not the intestine or neurons, seems to be the primary site of vang-1 function. Life span extension in vang-1 mutants depends on the insulin/IGF-1-like receptor DAF-2 and DAF-16/FoxO transcription factor. RNAi against the phase II detoxification transcription factor SKN-1/Nrf2 also reduced vang-1 life span that might be explained by gradual inhibition of insulin/IGF-1-like signaling in vang-1. This is the first time that a key player of the PCP pathway is shown to be involved in the insulin/IGF-1-like signaling dependent modulation of life span in C. elegans