Pertussis Toxin-sensitive Activation of Phospholipase C by the C5a and fMet-Leu-Phe Receptors

Signal transduction pathways that mediate C5a and fMet-Leu-Phe (fMLP)-induced pertussis toxin (PTx)-sensitive activation of phospholipase C (PLC) have been investigated using a cotransfection assay system in COS-7 cells. The abilities of the receptors for C5a and fMLP to activate PLC beta 2 and PLC beta 3 through the Gbeta gamma subunits of endogenous Gi proteins in COS-7 cells were tested because both PLC beta 2 and PLC beta 3 were shown to be activated by the beta gamma subunits of G proteins in in vitro reconstitution assays. Neither of the receptors can activate endogenous PLC beta 3 or recombinant PLC beta 3 in transfected COS-7 cells. However, both receptors can clearly activate PLC beta 2 in a PTx-sensitive manner, suggesting that the receptors may interact with endogenous PTx-sensitive G proteins and activate PLC beta 2 probably through the Gbeta gamma subunits. These findings were further corroborated by the results that PLC beta 3 could only be slightly activated by Gbeta 1gamma 1 or Gbeta 1gamma 5 in the cotransfection assay, whereas the Gbeta gamma subunits strongly activated PLC beta 2 under the same conditions. PLC beta 3 can be activated by Galpha q, Galpha 11, and Galpha 16 in the cotransfection assay. In addition, the Ggamma 2 and Ggamma 3 mutants with substitution of the C-terminal Cys residue by a Ser residue, which can inhibit wild type Gbeta gamma -mediated activation of PLC beta 2, were able to inhibit C5a or fMLP-mediated activation of PLC beta 2. These Ggamma mutants, however, showed little effect on m1-muscarinic receptor-mediated PLC activation, which is mediated by the Gq class of G proteins. These results all confirm that the Gbeta gamma subunits are involved in PLC beta 2 activation by the two chemoattractant receptors and suggest that in COS-7 cells activation of PLC beta 3 by Gbeta gamma may not be the primary pathway for the receptors

Evaluating Impacts of a Multilevel Resilience-Based Psychosocial Intervention on Mental Health of Children Affected by Parental HIV in China

Background: Children affected by parental HIV are commonly exposed to multiple risk factors, including parental illness and death, other traumatic life events, HIV stigma, and poverty, all of which in turn put them at elevated risk of experiencing poor mental health outcomes. Previous research has suggested the promise of psychosocial interventions in improving mental health for children affected by parental HIV through an integrated and multilevel resilience-based approach. However, there are few multilevel resilience-based interventions for this group, and the efficacy of such interventions on mental health outcomes has not been fully examined. Furthermore, very few studies have examined whether resilience-based interventions impact various sub-populations differently or explored the mechanisms through which such intervention effects occur. Therefore, the first aim of this dissertation research was to examine the short-term efficacy (e.g., up to 18 months) of the Child-Caregiver-Advocacy Resilience (ChildCARE) intervention, a multilevel resilience-based psychosocial intervention, on selected mental health outcomes (i.e., depressive symptoms, school anxiety, loneliness) among children affected by parental HIV, as well as testing the potential moderation roles of gender and age in the intervention effects. The second aim of this dissertation research was to examine whether the ChildCARE intervention would yield improvement in mental health beyond 18 months of follow up and whether emotional regulation and coping would act as the potential mechanisms of change through which the ChildCARE intervention improves mental health outcomes for these children. Methods: The ChildCARE intervention is a culturally tailored intervention developed for children affected by parental HIV in China, which consists of intervention components at three levels: child, caregiver, and community. The intervention was evaluated using a 4-arm community-based cluster randomized controlled trial with a sample of 790 children 6-17 years of age (51.6% boys) affected by parental HIV in a rural county in central China from 2012 to 2016. Children and their primary caregivers were randomly assigned by school clusters to a control condition or one of three intervention conditions (i.e., child-only, child + caregiver, child + caregiver + community). Of the three intervention conditions, children and caregivers assigned to the child-only condition were provided child intervention component only, those assigned to the child + caregiver condition were provided both child and caregiver intervention components, and children and caregivers assigned to the child + caregiver + community condition were provided all three intervention components. Data on depressive symptoms, school anxiety, loneliness, emotional regulation, coping, and demographic characteristics were collected from children via self-report at baseline and every six months over 36 months. Results: Overall, the ChildCARE intervention yielded some short-term improvements in depressive symptoms and loneliness, but these improvements were not sustained at 18 months or beyond for children affected by parental HIV. Older children (i.e., ≥ 12 years of age) benefited more from the intervention than their younger counterparts (i.e., \u3c 12 years). Mediation analyses further showed that the ChildCARE intervention yielded significant improvements in positive coping, but not emotional regulation or negative coping at 18 months, whereas changes in emotional regulation, positive coping, and negative coping were consistently associated with depressive symptoms, school anxiety, and loneliness. Conclusions: The findings in this research provide support for the benefits of the ChildCARE intervention on mental health outcomes but highlight the challenges in producing robust, long-term impacts for children affected by parental HIV in central China. This research also suggests the important roles of emotional regulation and coping in influencing mental health outcomes for these children

Co-infection of a hypovirulent isolate of Sclerotinia sclerotiorum with a new botybirnavirus and a strain of a mitovirus

BACKGROUND: Sclerotinia sclerotiorum, a notorious plant fungal pathogen, causes yield loss of many crops and vegetables, and is a natural host of a diverse viruses with positive-sense RNA (+ssRNA), negative-sense RNA (−ssRNA), double-stranded RNA (dsRNA), or DNA genomes. Mixed-infection with multiple related or unrelated mycoviruses is a common phenomenon in S. sclerotiorum. However, a single strain co-infected with dsRNA and + ssRNA viruses has not been reported in S. sclerotiorum. RESULTS: We report two unrelated viruses, Sclerotinia sclerotiorum botybirnavirus 2 (SsBRV2) with a bipartite dsRNA genome and Sclerotinia sclerotiorum mitovirus 4 (SsMV4/AH16) with a + ssRNA genome, which were originally detected in a single hypovirulent strain AH16 of S. sclerotiorum. SsMV4/AH16 has a typical genome of mitovirus and is a strain of mitovirus SsMV4. The genome of SsBRV2 consists of two separated dsRNA segments. The large dsRNA segment is 6159 bp in length and only has a single open reading frame (ORF) encoding a putative 1868-aa polyprotein with a conserved RNA dependent RNA polymerase (RdRp) domain. The small dsRNA segment is 5872 bp in length and encodes a putative 1778-aa protein. Phylogenetic analysis using RdRp conserved domain sequences revealed that SsBRV2 is phylogenetically related to the previously reported three bipartite viruses SsBRV1, Botrytis porri RNA virus 1 (BpRV1), and soybean leaf-associated botybirnavirus 1 (SlaBRV1). Electron microscopy demonstrated that SsBRV2 forms rigid spherical virions with a diameter of approximately 40 nm in infected mycelia. The virion of SsBRV2 was successfully introduced into a virus-free strain, which provides conclusive evidence that SsBRV2 confers hypovirulence on phytopathogenic fungus S. sclerotiorum. CONCLUSIONS: A bisegmented dsRNA virus (SsBRV2/AH16) and a nonsegmented + ssRNA virus (SsMV4/AH16) were characterized in a hypovirulent strain AH16 of S. sclerotiorum. SsMV4/AH16 is a strain of a reported mitovirus, whereas SsBRV2 is a new botybirnavirus. SsBRV2 is the causal agent of hypovirulence on S. sclerotiorum. Our findings supplied a first evidence that a single S. sclerotiorum strain is co-infected by dsRNA and + ssRNA mycoviruses. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12985-016-0550-2) contains supplementary material, which is available to authorized users

Histone H3 Lysine 9 Methyltransferase DIM5 Is Required for the Development and Virulence of Botrytis cinerea

Histone methylation is widely present in animals, plants and fungi, and the methylation modification of histone H3 has important biological functions. Methylation of Lys9 of histone H3 (H3K9) has been proven to regulate chromatin structure, gene silencing, transcriptional activation, plant metabolism and other processes. In this work, we investigated the functions of a H3K9 methyltransferase gene BcDIM5 in Botrytis cinerea, which contains a PreSET domain, a SET domain and a PostSET domain. Characterization of BcDIM5 knockout transformants showed that the hyphal growth rate and production of conidiophores and sclerotia were significantly reduced, while complementary transformation of BcDIM5 could restore the phenotypes to the levels of wild type. Pathogenicity assays revealed that BcDIM5 was essential for full virulence of B. cinerea. BcDIM5 knockout transformants exhibited decreased virulence, down-regulated expression of some pathogenic genes and drastically decreased H3K9 trimethylation level. However, knockout transformants of other two genes heterochromatin protein 1 (HP1) BcHP1 and DNA methyltransferase (DIM2) BcDIM2 did not exhibit significant change in the growth phenotype and virulence compared with the wild type. Our results indicate that H3K9 methyltransferase BcDIM5 is required for H3K9 trimethylation to regulate the development and virulence of B. cinerea

Cytoplasmic chromatin triggers inflammation in senescence and cancer

Chromatin is traditionally viewed as a nuclear entity that regulates gene expression and silencing. However, we recently discovered the presence of cytoplasmic chromatin fragments that pinch off from intact nuclei of primary cells during senescence, a form of terminal cell-cycle arrest associated with pro-inflammatory responses. The functional significance of chromatin in the cytoplasm is unclear. Here we show that cytoplasmic chromatin activates the innate immunity cytosolic DNA-sensing cGAS-STING (cyclic GMP-AMP synthase linked to stimulator of interferon genes) pathway, leading both to short-term inflammation to restrain activated oncogenes and to chronic inflammation that associates with tissue destruction and cancer. The cytoplasmic chromatin-cGAS-STING pathway promotes the senescence-associated secretory phenotype in primary human cells and in mice. Mice deficient in STING show impaired immuno-surveillance of oncogenic RAS and reduced tissue inflammation upon ionizing radiation. Furthermore, this pathway is activated in cancer cells, and correlates with pro-inflammatory gene expression in human cancers. Overall, our findings indicate that genomic DNA serves as a reservoir to initiate a pro-inflammatory pathway in the cytoplasm in senescence and cancer. Targeting the cytoplasmic chromatin-mediated pathway may hold promise in treating inflammation-related disorders

1. スポロトリコーシス5例(第443回千葉医学会例会 第16回千葉皮膚科臨床談話会)

A potential stable stem-loop structure in the 5’-terminal sequence (left) and a triple stem-loop structure in 3’-terminal sequences (right) were predicted with a RNA structure software. (PDF 60 kb

Проблемы формирования механизма мотивации персонала промышленного предприятия

Цель данной работы — углубление теоретических положений проблемы мотивации и разработка механизмов мотивации, стимулирования труда персонала промышленных предприятий в условиях формирования рыночных отношений.Исследованы теоретические и практические аспекты мотивации персонала, её структура и содержание. Разработаны критерии оценки эффективности систем мотивации персонала, модель формирования и мониторинга системы мотивации на основе уточненных факторов РичиМартина, трудовой концепции работника, построения «мотивационного профиля бюллетеня работника».Досліджені теоретичні і практичні аспекти мотивації персоналу, її структура і зміст. Розроблені критерії оцінки ефективності систем мотивації персоналу, модель формування і моніторингу системи мотивації на основі уточнених чинників Ричи-Мартіна, трудової концепції працівника, побудови «мотиваційного профілю бюлетеня працівника».The theoretical and practical aspects of motivation, its structure and content. Develop criteria for evaluating the effectiveness of staff motivation, a model of monitoring and incentive system based on the corrected factors Richie Martin, working the concept of worker, building a «motivational profile employee newsletter