MicroRNA 130a Regulates both Hepatitis C Virus and Hepatitis B Virus Replication through a Central Metabolic Pathway

ABSTRACT Hepatitis C virus (HCV) infection has been shown to regulate microRNA 130a (miR-130a) in patient biopsy specimens and in cultured cells. We sought to identify miR-130a target genes and to explore the mechanisms by which miR-130a regulates HCV and hepatitis B virus (HBV) replication. We used bioinformatics software, including miRanda, TargetScan, PITA, and RNAhybrid, to predict potential miR-130a target genes. miR-130a and its target genes were overexpressed or were knocked down by use of small interfering RNA (siRNA) or clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 guide RNA (gRNA). Selected gene mRNAs and their proteins, together with HCV replication in OR6 cells, HCV JFH1-infected Huh7.5.1 cells, and HCV JFH1-infected primary human hepatocytes (PHHs) and HBV replication in HepAD38 cells, HBV-infected NTCP-Huh7.5.1 cells, and HBV-infected PHHs, were measured by quantitative reverse transcription-PCR (qRT-PCR) and Western blotting, respectively. We selected 116 predicted target genes whose expression was related to viral pathogenesis or immunity for qPCR validation. Of these, the gene encoding pyruvate kinase in liver and red blood cell (PKLR) was confirmed to be regulated by miR-130a overexpression. miR-130a overexpression (via a mimic) knocked down PKLR mRNA and protein levels. A miR-130a inhibitor and gRNA increased PKLR expression, HCV replication, and HBV replication, while miR-130a gRNA and PKLR overexpression increased HCV and HBV replication. Supplemental pyruvate increased HCV and HBV replication and rescued the inhibition of HCV and HBV replication by the miR-130a mimic and PKLR knockdown. We concluded that miR-130a regulates HCV and HBV replication through its targeting of PKLR and subsequent pyruvate production. Our data provide novel insights into key metabolic enzymatic pathway steps regulated by miR-130a, including the steps involving PKLR and pyruvate, which are subverted by HCV and HBV replication. IMPORTANCE: We identified that miR-130a regulates the target gene PKLR and its subsequent effect on pyruvate production. Pyruvate is a key intermediate in several metabolic pathways, and we identified that pyruvate plays a key role in regulation of HCV and HBV replication. This previously unrecognized, miRNA-regulated antiviral mechanism has implications for the development of host-directed strategies to interrupt the viral life cycle and prevent establishment of persistent infection for HCV, HBV, and potentially other viral infections

69th Issue Info Kampus UiTM Sarawak Buletin : Januari 2015 / Professor Dato Dr. Jamil Hj Hamali... [et al.]

A very happy 2015 to UITM staff and students. I would like to express my deepest appreciation to the Chief Minister of Sarawak, Yang Amat Berhormat Dato Patinggi Tan Sri (Dr) Haji Adenan bin Haji Satem and the Vice - Chancellor of UITM,Yang Berbahagia Tan Sri Dato' Sri Professor Ir. Dr. Sahol Hamid Abu Bakar, FASc for the continues support given to UITM Sarawak.To the staff, i thank you all for your cooperation and dedication which have contributed to the tremendous development made by UITM Sarawak.In addition, i also encourage UITM Sarawak student to work hard and excel in their studies

De Novo Mutations in SLC1A2 and CACNA1A Are Important Causes of Epileptic Encephalopathies

Epileptic encephalopathies (EEs) are the most clinically important group of severe early-onset epilepsies. Next-generation sequencing has highlighted the crucial contribution of de novo mutations to the genetic architecture of EEs as well as to their underlying genetic heterogeneity. Our previous whole-exome sequencing study of 264 parent-child trios revealed more than 290 candidate genes in which only a single individual had a de novo variant. We sought to identify additional pathogenic variants in a subset (n = 27) of these genes via targeted sequencing in an unsolved cohort of 531 individuals with a diverse range of EEs. We report 17 individuals with pathogenic variants in seven of the 27 genes, defining a genetic etiology in 3.2% of this unsolved cohort. Our results provide definitive evidence that de novo mutations in SLC1A2 and CACNA1A cause specific EEs and expand the compendium of clinically relevant genotypes for GABRB3. We also identified EEs caused by genetic variants in ALG13, DNM1, and GNAO1 and report a mutation in IQSEC2. Notably, recurrent mutations accounted for 7/17 of the pathogenic variants identified. As a result of high-depth coverage, parental mosaicism was identified in two out of 14 cases tested with mutant allelic fractions of 5%–6% in the unaffected parents, carrying significant reproductive counseling implications. These results confirm that dysregulation in diverse cellular neuronal pathways causes EEs, and they will inform the diagnosis and management of individuals with these devastating disorders

Well-Child Care Clinical Practice Redesign at a Community Health Center

Background: Community health centers (CHCs) are a key element of the health care safety net for underserved children. They may be an ideal setting to create well-child care (WCC) clinical practice redesign to drastically improve WCC delivery. Objective: To examine the perspectives of clinical and administrative staff at a large, multisite urban CHC on alternative ways to deliver WCC services for low-income children aged 0 to 3 years. Methods: Eight semistructured interviews were conducted with 4 pediatric teams (each consisting of 1 pediatrician and 2 medical assistants) and 4 CHC executive/administrative staff (Medical Director, COO, CEO, and Nurse Supervisor). Discussions were recorded, transcribed, and analyzed using the constant comparative method of qualitative analysis. Salient themes included WCC delivery challenges and endorsed WCC clinical practice redesign solutions. Results: The 3 main WCC delivery challenges included long wait times due to insurance verification and intake paperwork, lack of time for parent education and sick visits due to WCC visit volume, and absence of a system to encourage physicians to use non–face-to-face communication with parents. To address WCC delivery challenges, CHC providers and administrators endorsed several options for clinical practice redesign in their setting. These included use of a health educator in a team-based model of care, a previsit tool for screening and surveillance, Web site health education, a structured system for non–face-to-face (eg, phone) parent communication, and group visits. Conclusion: CHC-specific strategies for WCC clinical practice redesign endorsed by a large, multisite safety net clinic may lead to more efficient, effective, and family-centered WCC for low-income populations

Parents views on engaging families of middle school students in obesity prevention and control in a multiethnic population.

INTRODUCTION: Overweight and obesity remain significant public health risks for youth in the United States, particularly among racial/ethnic minority groups. Efforts at obesity prevention and control have targeted youth and family members in diverse settings. Although involving parents in obesity prevention programs for youth may improve the potential of these programs, less is known about parents preferred methods of engagement, especially among racial/ethnic minority parents and parents whose primary language is not English. In this qualitative study, parents of middle-school-aged children were asked how best to engage their children in obesity prevention and control efforts. METHODS: We recruited 38 parents whose children attended Los Angeles middle schools to participate in focus groups. Two English-language focus groups with 14 parents of different racial/ethnic backgrounds and 2 Spanish language groups with 24 Latino parents were conducted from 2010 through 2011. We analyzed focus group transcripts by using content analysis using inductive and deductive techniques. RESULTS: Findings from focus groups confirmed that parents want to help their children avoid obesity but feel constrained in their ability to take action. Participants identified an overarching desire to become better parents as a potential incentive to engage in obesity prevention efforts. Parents advocated for family-focused approaches in obesity prevention programs, including family sports leagues and cooking classes. Most findings were consistent between language groups, but parents in the Spanish language groups cited language-related barriers. CONCLUSION: The development and testing of simple programs that are sustainable, community-based, and family-focused may empower families to address obesity prevention and control

Parents' views on engaging families of middle school students in obesity prevention and control in a multiethnic population.

IntroductionOverweight and obesity remain significant public health risks for youth in the United States, particularly among racial/ethnic minority groups. Efforts at obesity prevention and control have targeted youth and family members in diverse settings. Although involving parents in obesity prevention programs for youth may improve the potential of these programs, less is known about parents' preferred methods of engagement, especially among racial/ethnic minority parents and parents whose primary language is not English. In this qualitative study, parents of middle-school-aged children were asked how best to engage their children in obesity prevention and control efforts.MethodsWe recruited 38 parents whose children attended Los Angeles middle schools to participate in focus groups. Two English-language focus groups with 14 parents of different racial/ethnic backgrounds and 2 Spanish language groups with 24 Latino parents were conducted from 2010 through 2011. We analyzed focus group transcripts by using content analysis using inductive and deductive techniques.ResultsFindings from focus groups confirmed that parents want to help their children avoid obesity but feel constrained in their ability to take action. Participants identified an overarching desire to become better parents as a potential incentive to engage in obesity prevention efforts. Parents advocated for family-focused approaches in obesity prevention programs, including family sports leagues and cooking classes. Most findings were consistent between language groups, but parents in the Spanish language groups cited language-related barriers.ConclusionThe development and testing of simple programs that are sustainable, community-based, and family-focused may empower families to address obesity prevention and control

Parents' views on engaging families of middle school students in obesity prevention and control in a multiethnic population.

Overweight and obesity remain significant public health risks for youth in the United States, particularly among racial/ethnic minority groups. Efforts at obesity prevention and control have targeted youth and family members in diverse settings. Although involving parents in obesity prevention programs for youth may improve the potential of these programs, less is known about parents' preferred methods of engagement, especially among racial/ethnic minority parents and parents whose primary language is not English. In this qualitative study, parents of middle-school-aged children were asked how best to engage their children in obesity prevention and control efforts.We recruited 38 parents whose children attended Los Angeles middle schools to participate in focus groups. Two English-language focus groups with 14 parents of different racial/ethnic backgrounds and 2 Spanish language groups with 24 Latino parents were conducted from 2010 through 2011. We analyzed focus group transcripts by using content analysis using inductive and deductive techniques.Findings from focus groups confirmed that parents want to help their children avoid obesity but feel constrained in their ability to take action. Participants identified an overarching desire to become better parents as a potential incentive to engage in obesity prevention efforts. Parents advocated for family-focused approaches in obesity prevention programs, including family sports leagues and cooking classes. Most findings were consistent between language groups, but parents in the Spanish language groups cited language-related barriers.The development and testing of simple programs that are sustainable, community-based, and family-focused may empower families to address obesity prevention and control