ABSTRACT
AIM: A better understanding about the health care and support needs is beneficial to maintain the linkage between People Living with Human Immunodeficiency Virus (PLWH) and healthcare services. This study aims to explore the healthcare needs of PLWH from their perspectives. DESIGN: A qualitative descriptive study design was undertaken in July 2017 to June 2018. METHODS: Fourteen participants were recruited by purposive sampling technique. Face-to-face in-depth interview and focus-group discussion (FGD) were conducted and analysed using content analysis. RESULTS: Data revealed five themes, namely the needs to be free from stigma and discrimination, strengthen life spirit, have safe sexual practices, social support, and have accessible and affordable healthcare services. CONCLUSION: People Living with Human Immunodeficiency Virus have their own perspectives on their healthcare needs. Therefore, nurses and other healthcare providers need to explore, understand, and respond to the needs, and incorporate them into comprehensive and holistic care.
Subject(s)
HIV Infections , Delivery of Health Care , HIV , Humans , Qualitative Research , Social StigmaABSTRACT
The involvement of host proteins in the replication and transcription of viral RNA is a poorly understood area for many RNA viruses. For coronaviruses, it was long speculated that replication of the giant RNA genome and transcription of multiple subgenomic mRNA species by a unique discontinuous transcription mechanism may require host cofactors. To search for such cellular proteins, yeast two-hybrid screening was carried out by using the nonstructural protein 14 (nsp14) from the coronavirus infectious bronchitis virus (IBV) as a bait protein, leading to the identification of DDX1, a cellular RNA helicase in the DExD/H helicase family, as a potential interacting partner. This interaction was subsequently confirmed by coimmunoprecipitation assays with cells coexpressing the two proteins and with IBV-infected cells. Furthermore, the endogenous DDX1 protein was found to be relocated from the nucleus to the cytoplasm in IBV-infected cells. In addition to its interaction with IBV nsp14, DDX1 could also interact with the nsp14 protein from severe acute respiratory syndrome coronavirus (SARS-CoV), suggesting that interaction with DDX1 may be a general feature of coronavirus nsp14. The interacting domains were mapped to the C-terminal region of DDX1 containing motifs V and VI and to the N-terminal portion of nsp14. Manipulation of DDX1 expression, either by small interfering RNA-induced knockdown or by overexpression of a mutant DDX1 protein, confirmed that this interaction may enhance IBV replication. This study reveals that DDX1 contributes to efficient coronavirus replication in cell culture.
Subject(s)
Coronavirus Infections/metabolism , DEAD-box RNA Helicases/metabolism , Infectious bronchitis virus/physiology , Viral Nonstructural Proteins/metabolism , Virus Replication , Animals , Cell Nucleus/metabolism , Chlorocebus aethiops , Coronavirus Infections/genetics , Coronavirus Infections/virology , Cytoplasm/metabolism , DEAD-box RNA Helicases/genetics , HeLa Cells , Humans , Infectious bronchitis virus/genetics , Protein Binding , Protein Transport , Two-Hybrid System Techniques , Vero Cells , Viral Nonstructural Proteins/geneticsABSTRACT
Open reading frame 89 (ORF89) is one of the three genes that are believed to be involved in the latent infection of white spot syndrome virus (WSSV). Here, we report the structure and functional characterization of ORF89. cDNA sequencing, 5' RLM-RACE, and 3' RLM-RACE showed that ORF89 gene is transcribed into an unspliced mRNA of 4436 nucleotides, which is predicted to encode a protein of 1437 amino acids. ORF89 expressed an approximately 165-kDa protein in Sf9 cells that localized in the nucleus. Amino acids 678-683 were found to be essential for nuclear localization. Cotransfection assays demonstrated that ORF89 protein repressed its own promoter as well as those of a protein kinase and the thymidine-thymidylate kinase genes of WSSV. SYBR Green real-time PCR indicated that the repression occurred at the transcriptional level.
Subject(s)
DNA Viruses/genetics , Genes, Viral , Open Reading Frames , Penaeidae/virology , Virus Latency/genetics , Animals , Base Sequence , Cell Line , Molecular Sequence Data , Nuclear Localization Signals , Promoter Regions, Genetic , Protein Kinases/genetics , RNA, Messenger/analysis , Spodoptera , Thymidine Kinase/geneticsABSTRACT
To investigate whether specific-pathogen-free (SPF) shrimps are asymptomatic carriers of white spot syndrome virus (WSSV), we used a WSSV-specific DNA microarray to measure WSSV gene expression in SPF and WSSV-infected shrimps. Three WSSV genes were found to be relatively highly expressed in SPF shrimps. Reverse transcription-PCR using nested primers as well as real-time detection confirmed that these genes have no detectable counterparts in GenBank; structural analysis of the putative proteins revealed helix-loop-helix and leucine zipper motifs. Viral sequences could be PCR amplified from genomic DNA of SPF shrimp, further supporting the suggestion that these shrimps are asymptomatic carriers.
