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1.
Artigo em Inglês | MEDLINE | ID: mdl-38791777

RESUMO

Lead is an established neurotoxicant, and it has known associations with adverse neurodevelopmental and reproductive outcomes. Exposure to lead at any level is unsafe, and the United States (US) has enacted various federal and state legislations to regulate lead levels in drinking water in K-12 schools and childcare facilities; however, no regulations exist for higher education settings. Upon the discovery of lead in drinking water fixtures in the University of North Carolina at Chapel Hill (UNC-CH) campus, a cross-campus water testing network and sampling plan was developed and deployed. The campaign was based on the US Environmental Protection Agency's (EPA) 3Ts (Training, Testing, and Taking Action) guidance. The seven-month campaign involved 5954 tests on 3825 drinking water fixtures across 265 buildings. A total of 502 (8.43%) tests showed lead above the limit of detection (1 part per billion, ppb), which represented 422 (11.03%) fixtures. Fewer than 1.5% of the tests were above the EPA action level for public water systems (15 ppb). In conclusion, systematic testing of all the fixtures across campus was required to identify localized contamination, and each entity in the cross-campus network undertook necessary roles to generate a successful testing campaign. UNC-CH established preventative measures to test drinking water fixtures every three years, which provide a framework for other higher education institutions in responding to lead contamination.


Assuntos
Água Potável , Chumbo , Chumbo/análise , Água Potável/análise , Água Potável/química , Universidades , North Carolina , Poluentes Químicos da Água/análise , Humanos , Monitoramento Ambiental , Estados Unidos , United States Environmental Protection Agency
2.
Emerg Microbes Infect ; 9(1): 124-128, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-31913767

RESUMO

A serological survey of 2,430 archived serum samples collected between 1997 and 2012 was conducted to retrospectively determine the prevalence of Marburg virus in five African countries. Serum samples were screened for neutralizing antibodies in a pseudotype micro-neutralization assay and confirmed by enzyme-linked immunosorbent assay (ELISA). Surprisingly, a seroprevalence for Marburg virus of 7.5 and 6.3% was found in Cameroon and Ghana, respectively, suggesting the circulation of filoviruses or related viruses outside of known endemic areas that remain undetected by current surveillance efforts. However, due to the lack of validated assays and appropriate positive controls, these results must be considered preliminary.


Assuntos
Anticorpos Antivirais/sangue , Filoviridae/imunologia , Doença do Vírus de Marburg/sangue , Doença do Vírus de Marburg/epidemiologia , Marburgvirus/imunologia , Animais , Camarões/epidemiologia , Ensaio de Imunoadsorção Enzimática , Filoviridae/genética , Infecções por Filoviridae/sangue , Infecções por Filoviridae/epidemiologia , Infecções por Filoviridae/virologia , Gana/epidemiologia , Humanos , Doença do Vírus de Marburg/virologia , Marburgvirus/genética , Estudos Retrospectivos , Estudos Soroepidemiológicos
3.
Front Microbiol ; 10: 1581, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31379769

RESUMO

Acinetobacter baumannii (Ab) is one of the most treacherous pathogens among those causing hospital-acquired pneumonia (HAP). A. baumannii possesses an adaptable physiology, seen not only in its antibiotic resistance and virulence phenotypes but also in its metabolic versatility. In this study, we observed that A. baumannii undergoes global transcriptional changes in response to human pleural fluid (PF), a key host-derived environmental signal. Differential gene expression analyses combined with experimental approaches revealed changes in A. baumannii metabolism, affecting cytotoxicity, persistence, bacterial killing, and chemotaxis. Over 1,220 genes representing 55% of the differentially expressed transcriptomic data corresponded to metabolic processes, including the upregulation of glutamate, short chain fatty acid, and styrene metabolism. We observed an upregulation by 1.83- and 2.61-fold of the pyruvate dehydrogenase complex subunits E3 and E2, respectively. We also found that pyruvate (PYR), in conjunction with PF, triggers an A. baumannii pathogenic behavior that adversely impacts human epithelial cell viability. Interestingly, PF also amplified A. baumannii cytotoxicity against murine macrophages, suggesting an immune evasion strategy implemented by A. baumannii. Moreover, we uncovered opposing metabolic strategies dependent on the degree of pathogenicity of the strains, where less pathogenic strains demonstrated greater utilization of PYR to promote persister formation in the presence of PF. Additionally, our transcriptomic analysis and growth studies of A. baumannii suggest the existence of an alternative phenylalanine (PA) catabolic route independent of the phenylacetic acid pathway, which converts PA to phenylpyruvate (PP) and shuttles intermediates into styrene metabolism. This alternative route promoted a neutrophil-evasive state, as PF-induced degradation of PP significantly reduced overall human neutrophil chemotaxis in ex vivo chemotactic assays. Taken together, these data highlight A. baumannii pathoadaptabililty in response to host signals and provide further insight into the role of bacterial metabolism in virulence traits, antibiotic persistence strategies, and host innate immune evasion.

4.
Emerg Infect Dis ; 25(5): 911-918, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-31002071

RESUMO

We conducted a serologic survey of 2,430 serum samples collected during 1997-2012 for various studies to determine the prevalence of the hemorrhagic fever virus Ebola virus (EBOV) in equatorial Africa. We screened serum samples for neutralizing antibodies by using a pseudotype microneutralization assay and a newly developed luciferase immunoprecipitation system assay. Specimens seroreactive for EBOV were confirmed by using an ELISA. Our results suggest a serologic prevalence of 2%-3.5% in the Republic of the Congo and the Democratic Republic of the Congo, which have reported outbreaks of infection with EBOV. In addition we detected a seroprevalence of 1.3% in southern Cameroon, which indicated a low risk for exposure in this region.


Assuntos
Ebolavirus , Doença pelo Vírus Ebola/epidemiologia , África Central/epidemiologia , Anticorpos Antivirais/sangue , Ebolavirus/imunologia , Ensaio de Imunoadsorção Enzimática , Células HEK293 , Doença pelo Vírus Ebola/sangue , Humanos , Imunoprecipitação , Nucleoproteínas/imunologia , Estudos Soroepidemiológicos , Proteínas do Core Viral/imunologia , Proteínas do Envelope Viral/imunologia
5.
Biochem Biophys Res Commun ; 505(3): 726-732, 2018 11 02.
Artigo em Inglês | MEDLINE | ID: mdl-30292413

RESUMO

The production of reactive oxygen species (ROS) is a prominent response to infection among innate immune cells such as macrophages and neutrophils. To better understand the relationship between antimicrobial and regulatory functions of blood cell ROS, we have characterized the ROS response to infection in Drosophila hemocytes. Using fluorescent probes, we find a biphasic hemocyte ROS response to bacterial infection. In the first hour, virtually all hemocytes generate a transient ROS signal, with nonphagocytic cells including prohemocytes and crystal cells displaying exceptionally strong responses. A distinct, and more delayed ROS response starting at 90 min is primarily within cells that have engulfed bacteria, and is sustained for several hours. The early response has a clear regulatory function, as dampening or intensifying the intracellular ROS level has profound effects on plasmatocyte activation. In addition, ROS are necessary and sufficient to activate JNK signalling in crystal cells, and to promote JNK-dependent crystal cell rupture. These findings indicate that Drosophila will be a promising model in which to dissect the mechanisms of ROS stimulation of immune activation.


Assuntos
Escherichia coli/imunologia , Hemócitos/imunologia , Macrófagos/imunologia , Espécies Reativas de Oxigênio/imunologia , Animais , Drosophila , Escherichia coli/genética , Escherichia coli/fisiologia , Hemócitos/metabolismo , Hemócitos/microbiologia , Interações Hospedeiro-Patógeno/imunologia , Larva/imunologia , Larva/metabolismo , Larva/microbiologia , Proteínas Luminescentes/genética , Proteínas Luminescentes/metabolismo , Sistema de Sinalização das MAP Quinases/imunologia , Macrófagos/metabolismo , Macrófagos/microbiologia , Microscopia de Fluorescência , Fagocitose/imunologia , Espécies Reativas de Oxigênio/metabolismo
6.
Mol Biol Cell ; 29(18): 2137-2147, 2018 09 01.
Artigo em Inglês | MEDLINE | ID: mdl-29995573

RESUMO

Wound reepithelialization is an evolutionarily conserved process in which skin cells migrate as sheets to heal the breach and is critical to prevent infection but impaired in chronic wounds. Integrin heterodimers mediate attachment between epithelia and underlying extracellular matrix and also act in large signaling complexes. The complexity of the mammalian wound environment and evident redundancy among integrins has impeded determination of their specific contributions to reepithelialization. Taking advantage of the genetic tools and smaller number of integrins in Drosophila, we undertook a systematic in vivo analysis of integrin requirements in the reepithelialization of skin wounds in the larva. We identify αPS2-ßPS and αPS3-ßPS as the crucial integrin dimers and talin as the only integrin adhesion component required for reepithelialization. The integrins rapidly accumulate in a JNK-dependent manner in a few rows of cells surrounding a wound. Intriguingly, the integrins localize to the distal margin in these cells, instead of the frontal or lamellipodial distribution expected for proteins providing traction and recruit nonmuscle myosin II to the same location. These findings indicate that signaling roles of integrins may be important for epithelial polarization around wounds and lay the groundwork for using Drosophila to better understand integrin contributions to reepithelialization.


Assuntos
Proteínas de Drosophila/metabolismo , Cadeias alfa de Integrinas/metabolismo , Integrinas/fisiologia , Cicatrização/fisiologia , Animais , Movimento Celular , Drosophila/metabolismo , Proteínas de Drosophila/fisiologia , Epitélio/metabolismo , Epitélio/fisiologia , Matriz Extracelular , Cadeias alfa de Integrinas/fisiologia , Integrinas/metabolismo , Larva , Morfogênese , Fenótipo , Transdução de Sinais , Talina/metabolismo
7.
PLoS One ; 12(6): e0178988, 2017.
Artigo em Inglês | MEDLINE | ID: mdl-28582431

RESUMO

BACKGROUND: The prevalence of chronic Hepatitis B Virus (HBV) infection is 2-4% in the Pakistani population, defining Pakistan as an intermediate prevalence country. In this study, hepatitis B surface antigen (HBsAg) reactive blood donations were screened using a combination of serological and molecular methods to identify immune escape HBV mutant strains and to determine the HBV genotypes and subtypes present in Pakistan. METHODS: Blood donations were collected at the Armed Forces Institute of Transfusion (AFIT) located in northern Pakistan and the Hussaini Blood Bank (HBB) located in the south. From 2009 to 2013 a total of 706,575 donations were screened with 2.04% (14,409) HBsAg reactive. A total of 2055 HBsAg reactive specimens, were collected and screened using a monoclonal antibody based research assay to identify immune escape mutants followed by PCR amplification and DNA sequencing to identify the mutation present. DNA sequences obtained from 192 specimens, including mutant candidates and wild type strains, were analyzed for escape mutations, genotype, and HBsAg subtype. RESULTS: Mutations were identified in approximately 14% of HBsAg reactive donations. Mutations at HBsAg amino acid positions 143-145 are the most common (46%) with the mutation serine 143 to leucine the most frequently occurring change (28%). While regional differences were observed, the most prevalent HBV strains are subgenotypes of D with subgenotype D1/subtype ayw2 accounting for the majority of infections; 90.2% at AFIT and 52.5% at HBB. CONCLUSIONS: The high frequency of immune escape HBV mutants in HBV infected Pakistani blood donors highlights the need for more studies into the prevalence of escape mutants. Differences between vaccinated and unvaccinated populations, the correlation of escape mutant frequency with genotype, and impact of escape mutations in different genotype backgrounds on the performance of commercially available HBsAg assays represent avenues for further investigation.


Assuntos
Doadores de Sangue/estatística & dados numéricos , DNA Viral/genética , Genótipo , Antígenos de Superfície da Hepatite B/genética , Vírus da Hepatite B/genética , Mutação , Adolescente , Adulto , Idoso , Bancos de Sangue/organização & administração , Transfusão de Sangue , DNA Viral/imunologia , Seleção do Doador/estatística & dados numéricos , Feminino , Hepatite B/diagnóstico , Hepatite B/imunologia , Hepatite B/virologia , Antígenos de Superfície da Hepatite B/análise , Antígenos de Superfície da Hepatite B/imunologia , Vírus da Hepatite B/classificação , Vírus da Hepatite B/imunologia , Vírus da Hepatite B/isolamento & purificação , Humanos , Evasão da Resposta Imune , Masculino , Pessoa de Meia-Idade , Paquistão
8.
J Virol ; 91(6)2017 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-28077647

RESUMO

As the epidemiological epicenter of the human immunodeficiency virus (HIV) pandemic, the Democratic Republic of the Congo (DRC) is a reservoir of circulating HIV strains exhibiting high levels of diversity and recombination. In this study, we characterized HIV specimens collected in two rural areas of the DRC between 2001 and 2003 to identify rare strains of HIV. The env gp41 region was sequenced and characterized for 172 HIV-positive specimens. The env sequences were predominantly subtype A (43.02%), but 7 other subtypes (33.14%), 20 circulating recombinant forms (CRFs; 11.63%), and 20 unclassified (11.63%) sequences were also found. Of the rare and unclassified subtypes, 18 specimens were selected for next-generation sequencing (NGS) by a modified HIV-switching mechanism at the 5' end of the RNA template (SMART) method to obtain full-genome sequences. NGS produced 14 new complete genomes, which included pure subtype C (n = 2), D (n = 1), F1 (n = 1), H (n = 3), and J (n = 1) genomes. The two subtype C genomes and one of the subtype H genomes branched basal to their respective subtype branches but had no evidence of recombination. The remaining 6 genomes were complex recombinants of 2 or more subtypes, including subtypes A1, F, G, H, J, and K and unclassified fragments, including one subtype CRF25 isolate, which branched basal to all CRF25 references. Notably, all recombinant subtype H fragments branched basal to the H clade. Spatial-geographical analysis indicated that the diverse sequences identified here did not expand globally. The full-genome and subgenomic sequences identified in our study population significantly increase the documented diversity of the strains involved in the continually evolving HIV-1 pandemic.IMPORTANCE Very little is known about the ancestral HIV-1 strains that founded the global pandemic, and very few complete genome sequences are available from patients in the Congo Basin, where HIV-1 expanded early in the global pandemic. By sequencing a subgenomic fragment of the HIV-1 envelope from study participants in the DRC, we identified rare variants for complete genome sequencing. The basal branching of some of the complete genome sequences that we recovered suggests that these strains are more closely related to ancestral HIV-1 strains than to previously reported strains and is evidence that the local diversification of HIV in the DRC continues to outpace the diversity of global strains decades after the emergence of the pandemic.


Assuntos
Variação Genética , Infecções por HIV/virologia , HIV-1/classificação , HIV-1/genética , Análise por Conglomerados , República Democrática do Congo/epidemiologia , Genoma Viral , Genótipo , Proteína gp41 do Envelope de HIV/genética , Infecções por HIV/epidemiologia , HIV-1/isolamento & purificação , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Epidemiologia Molecular , População Rural , Homologia de Sequência
9.
Medsurg Nurs ; 26(2): 99-104, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30304589

RESUMO

Hip and knee arthroplasty are common 'surgical procedures. Enhanced recovery programs help reduce length of stay with good patient outcomes and satisfaction. An audit is described showing improved pain management, reduced nausea and vomiting, and quicker recovery through an interprofessional approach.


Assuntos
Artroplastia de Quadril/normas , Artroplastia do Joelho/normas , Enfermagem Perioperatória/normas , Enfermagem em Pós-Anestésico/normas , Guias de Prática Clínica como Assunto , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade
10.
AIDS Res Hum Retroviruses ; 33(4): 347-352, 2017 04.
Artigo em Inglês | MEDLINE | ID: mdl-27758113

RESUMO

HIV-2 exhibits a natural history of infection distinct from HIV-1. Primarily found in West Africa and in only 10%-20% of HIV infections in this region, patients with HIV-2 typically exhibit a slower progression to AIDS, lower viral loads, and decreased rates of transmission. Here, we used next-generation sequencing to determine the sequence and phylogenetic classification of nine HIV-2 genomes. We identified a patient with a series of mutations in an invariant cytotoxic lymphocyte (CTL)-restricted gag epitope required for retroviral structure and replication and implicated in long-term nonprogression to AIDS. The presence of wild-type sequence argues these mutations are involved in immune escape, whereas its reversion to a sequence seen only in the sooty mangabey reservoir suggests an alternate means of controlling infection. Surveillance and molecular characterization of circulating strains are essential for continued development of monitoring tools and may provide greater insight into the reduced pathogenicity of HIV-2.


Assuntos
Epitopos de Linfócito T/genética , Infecções por HIV/virologia , HIV-2/classificação , HIV-2/genética , Mutação de Sentido Incorreto , África Ocidental , Monitoramento Epidemiológico , Sobreviventes de Longo Prazo ao HIV , HIV-2/isolamento & purificação , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Epidemiologia Molecular , Produtos do Gene gag do Vírus da Imunodeficiência Humana/genética
11.
PLoS One ; 11(12): e0167411, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-27997536

RESUMO

We conducted a literature review of reported temperature, salinity, pH, depth and oxygen preferences and thresholds of important marine species found in the Gulf of St. Lawrence and Scotian Shelf region. We classified 54 identified fishes and macroinvertebrates as important either because they support a commercial fishery, have threatened or at risk status, or meet one of the following criteria: bycatch, baitfish, invasive, vagrant, important for ecosystem energy transfer, or predators or prey of the above species. The compiled data allow an assessment of species-level impacts including physiological stress and mortality given predictions of future ocean physical and biogeochemical conditions. If an observed, multi-decadal oxygen trend on the central Scotian Shelf continues, a number of species will lose favorable oxygen conditions, experience oxygen-stress, or disappear due to insufficient oxygen in the coming half-century. Projected regional trends and natural variability are both large, and natural variability will act to alternately amplify and dampen anthropogenic changes. When estimates of variability are included with the trend, species encounter unfavourable oxygen conditions decades sooner. Finally, temperature and oxygen thresholds of adult Atlantic wolffish (Anarhichas lupus) and adult Atlantic cod (Gadus morhua) are assessed in the context of a potential future scenario derived from high-resolution ocean models for the central Scotian Shelf.


Assuntos
Organismos Aquáticos/fisiologia , Mudança Climática , Gadus morhua/fisiologia , Modelos Biológicos , Consumo de Oxigênio/fisiologia , Animais , Escócia
12.
J Clin Microbiol ; 54(4): 868-82, 2016 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-26699702

RESUMO

Molecular surveillance is essential to monitor HIV diversity and track emerging strains. We have developed a universal library preparation method (HIV-SMART [i.e.,switchingmechanismat 5' end ofRNAtranscript]) for next-generation sequencing that harnesses the specificity of HIV-directed priming to enable full genome characterization of all HIV-1 groups (M, N, O, and P) and HIV-2. Broad application of the HIV-SMART approach was demonstrated using a panel of diverse cell-cultured virus isolates. HIV-1 non-subtype B-infected clinical specimens from Cameroon were then used to optimize the protocol to sequence directly from plasma. When multiplexing 8 or more libraries per MiSeq run, full genome coverage at a median ∼2,000× depth was routinely obtained for either sample type. The method reproducibly generated the same consensus sequence, consistently identified viral sequence heterogeneity present in specimens, and at viral loads of ≤4.5 log copies/ml yielded sufficient coverage to permit strain classification. HIV-SMART provides an unparalleled opportunity to identify diverse HIV strains in patient specimens and to determine phylogenetic classification based on the entire viral genome. Easily adapted to sequence any RNA virus, this technology illustrates the utility of next-generation sequencing (NGS) for viral characterization and surveillance.


Assuntos
Genoma Viral , Infecções por HIV/virologia , HIV/classificação , HIV/isolamento & purificação , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Plasma/virologia , Análise de Sequência de DNA/métodos , Camarões , Genótipo , HIV/genética , Humanos , Filogenia
13.
PLoS Pathog ; 11(12): e1005325, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26658760

RESUMO

Hepatitis C virus (HCV) and human pegivirus (HPgV), formerly GBV-C, are the only known human viruses in the Hepacivirus and Pegivirus genera, respectively, of the family Flaviviridae. We present the discovery of a second pegivirus, provisionally designated human pegivirus 2 (HPgV-2), by next-generation sequencing of plasma from an HCV-infected patient with multiple bloodborne exposures who died from sepsis of unknown etiology. HPgV-2 is highly divergent, situated on a deep phylogenetic branch in a clade that includes rodent and bat pegiviruses, with which it shares <32% amino acid identity. Molecular and serological tools were developed and validated for high-throughput screening of plasma samples, and a panel of 3 independent serological markers strongly correlated antibody responses with viral RNA positivity (99.9% negative predictive value). Discovery of 11 additional RNA-positive samples from a total of 2440 screened (0.45%) revealed 93-94% nucleotide identity between HPgV-2 strains. All 12 HPgV-2 RNA-positive cases were identified in individuals also testing positive for HCV RNA (12 of 983; 1.22%), including 2 samples co-infected with HIV, but HPgV-2 RNA was not detected in non-HCV-infected individuals (p<0.0001), including those singly infected by HIV (p = 0.0075) or HBV (p = 0.0077), nor in volunteer blood donors (p = 0.0082). Nine of the 12 (75%) HPgV-2 RNA positive samples were reactive for antibodies to viral serologic markers, whereas only 28 of 2,429 (1.15%) HPgV-2 RNA negative samples were seropositive. Longitudinal sampling in two individuals revealed that active HPgV-2 infection can persist in blood for at least 7 weeks, despite the presence of virus-specific antibodies. One individual harboring both HPgV-2 and HCV RNA was found to be seronegative for both viruses, suggesting a high likelihood of simultaneous acquisition of HCV and HPgV-2 infection from an acute co-transmission event. Taken together, our results indicate that HPgV-2 is a novel bloodborne infectious virus of humans and likely transmitted via the parenteral route.


Assuntos
Infecções por Flaviviridae/virologia , Vírus GB C/genética , Hepacivirus/genética , Hepatite C/virologia , Hepatite Viral Humana/virologia , Sequência de Bases , Coinfecção/genética , Coinfecção/virologia , Feminino , Infecções por Flaviviridae/genética , Hepatite C/genética , Hepatite Viral Humana/genética , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa
14.
PLoS One ; 10(11): e0141723, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26599538

RESUMO

Given the dynamic changes in HIV-1 complexity and diversity, next-generation sequencing (NGS) has the potential to revolutionize strategies for effective HIV global surveillance. In this study, we explore the utility of metagenomic NGS to characterize divergent strains of HIV-1 and to simultaneously screen for other co-infecting viruses. Thirty-five HIV-1-infected Cameroonian blood donor specimens with viral loads of >4.4 log10 copies/ml were selected to include a diverse representation of group M strains. Random-primed NGS libraries, prepared from plasma specimens, resulted in greater than 90% genome coverage for 88% of specimens. Correct subtype designations based on NGS were concordant with sub-region PCR data in 31 of 35 (89%) cases. Complete genomes were assembled for 25 strains, including circulating recombinant forms with relatively limited data available (7 CRF11_cpx, 2 CRF13_cpx, 1 CRF18_cpx, and 1 CRF37_cpx), as well as 9 unique recombinant forms. HPgV (formerly designated GBV-C) co-infection was detected in 9 of 35 (25%) specimens, of which eight specimens yielded complete genomes. The recovered HPgV genomes formed a diverse cluster with genotype 1 sequences previously reported from Ghana, Uganda, and Japan. The extensive genome coverage obtained by NGS improved accuracy and confidence in phylogenetic classification of the HIV-1 strains present in the study population relative to conventional sub-region PCR. In addition, these data demonstrate the potential for metagenomic analysis to be used for routine characterization of HIV-1 and identification of other viral co-infections.


Assuntos
Vírus GB C/genética , Variação Genética , HIV-1/genética , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Metagenômica , Doadores de Sangue , Camarões , Coinfecção/genética , Genoma Viral , Genótipo , Humanos , Dados de Sequência Molecular , Filogenia , Recombinação Genética
15.
AIDS Res Hum Retroviruses ; 31(4): 412-20, 2015 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-25492033

RESUMO

By the end of 2012, more than 6.1 million people were infected with HIV-1 in South Africa. Subtype C was responsible for the majority of these infections and more than 300 near full-length genomes (NFLGs) have been published. Currently very few non-subtype C isolates have been identified and characterized within the country, particularly full genome non-C isolates. Seven patients from the Tygerberg Virology (TV) cohort were previously identified as possible non-C subtypes and were selected for further analyses. RNA was isolated from five individuals (TV047, TV096, TV101, TV218, and TV546) and DNA from TV016 and TV1057. The NFLGs of these samples were amplified in overlapping fragments and sequenced. Online subtyping tools REGA version 3 and jpHMM were used to screen for subtypes and recombinants. Maximum likelihood (ML) phylogenetic analysis (phyML) was used to infer subtypes and SimPlot was used to confirm possible intersubtype recombinants. We identified three subtype B (TV016, TV047, and TV1057) isolates, one subtype A1 (TV096), one subtype G (TV546), one unique AD (TV101), and one unique AC (TV218) recombinant form. This is the first NFLG of subtype G that has been described in South Africa. The subtype B sequences described also increased the NFLG subtype B sequences in Africa from three to six. There is a need for more NFLG sequences, as partial HIV-1 sequences may underrepresent viral recombinant forms. It is also necessary to continue monitoring the evolution and spread of HIV-1 in South Africa, because understanding viral diversity may play an important role in HIV-1 prevention strategies.


Assuntos
Genoma Viral , Genótipo , Infecções por HIV/virologia , HIV-1/classificação , HIV-1/genética , Análise de Sequência de DNA , Adulto , Análise por Conglomerados , Feminino , HIV-1/isolamento & purificação , Humanos , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Filogenia , Recombinação Genética , Homologia de Sequência , África do Sul
17.
Aust J Rural Health ; 21(2): 72-9, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-23586568

RESUMO

OBJECTIVE: High levels of alcohol-related harm are a salient feature of many rural communities in Australia. General practitioners (GPs) are uniquely placed to identify and treat patients with harmful alcohol use in remote settings, yet corresponding opportunities for education in effective brief psychological interventions for harmful alcohol use are limited. This study piloted a training model for alcohol screening and brief intervention for GPs working in Kalgoorlie-Boulder, a remote Western Australian community facing significant alcohol-related problems. DESIGN: Observational pilot study. SETTING: Primary care. MAIN OUTCOME MEASURE(S): Perceived role in responding to harmful alcohol use, and confidence and knowledge of alcohol screening and brief intervention; satisfaction with a short training session focused on alcohol screening and brief intervention; and impact of training on implementation of screening and brief intervention for harmful alcohol use. RESULTS: Fifty per cent of GPs took up the training opportunity. GPs recognised their professional responsibility for conducting brief intervention but reported comparatively lower confidence and skills in implementing screening and intervention prior to training. The training improved knowledge and confidence in conducting alcohol screening and brief intervention. All GPs increased their frequency of alcohol screening, and 88% of GPs reported increasing the frequency of brief intervention at 6 months. CONCLUSIONS: Preliminary findings suggest that among participating GPs, subsequent compliance with identification and management of harmful alcohol use was improved. Further work examining methods to improve rural and remote GP participation in alcohol-related harm prevention training is required, as the potential impact on communities with disproportionately high alcohol-related difficulties is significant.


Assuntos
Alcoolismo/diagnóstico , Clínicos Gerais/educação , Entrevista Motivacional/métodos , População Rural , Alcoolismo/terapia , Humanos , Projetos Piloto , Austrália Ocidental
18.
J Clin Virol ; 57(2): 169-72, 2013 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-23485348

RESUMO

BACKGROUND: HIV antigen/antibody (Ag/Ab) combination assays represent a significant advancement in assays used for diagnosing HIV infection based on their ability to detect acute and chronic infections. During acute HIV infection (AHI), detection depends on assay sensitivity for p24 Ag. OBJECTIVE: To directly compare the Ag sensitivity of the ARCHITECT(®) HIV Ag/Ab Combo assay to RNA viral load using cell culture supernatants of virus isolates. HIV-1 isolates allow correlation in the total absence of an antibody response to infection and across genetically diverse HIV-1 group M strains. METHODS: Thirty-five HIV-1 isolates comprising subtypes A-D, F and G, CRF01_AE, CRF02_AG, and unique recombinant forms were evaluated. Cell-free culture supernatant for each isolate was diluted to four levels and tested in the HIV Combo assay to determine a signal to cutoff ratio and the RealTime(®) HIV-1 assay to quantify RNA. The RNA copies/mL at the HIV Combo assay cutoff was determined. RESULTS: The median RNA copies/mL at the HIV Combo assay cutoff was 57,900 for individual virus isolates (range 26,440-102,400). A single plot of all the data gave a value of 58,500RNA copies/mL. An analysis of data published for acute HIV infection in human subjects gave a similar result; HIV Combo detected 97% of AHIs with RNA copies/mL > 30,700. CONCLUSIONS: Based on analysis of virus isolates, the ARCHITECT HIV Combo assay can detect p24 Ag when RNA is above approximately 58,000copies/mL. The correlation of viral load and Ag sensitivity was consistent across genetically diverse HIV-1 group M strains.


Assuntos
Anticorpos Anti-HIV/sangue , Proteína do Núcleo p24 do HIV , Infecções por HIV/diagnóstico , HIV-1/genética , RNA Viral/sangue , Recombinação Genética , Carga Viral , Doença Aguda , Feminino , Variação Genética , Proteína do Núcleo p24 do HIV/sangue , Proteína do Núcleo p24 do HIV/imunologia , Infecções por HIV/imunologia , Infecções por HIV/virologia , HIV-1/classificação , HIV-1/imunologia , Humanos , Masculino , RNA Viral/genética , Kit de Reagentes para Diagnóstico , Sensibilidade e Especificidade
19.
J Virol ; 86(19): 10505-16, 2012 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-22811521

RESUMO

Although HLA-B*57 (B57) is associated with slow progression to disease following HIV-1 infection, B57 heterozygotes display a wide spectrum of outcomes, including rapid progression, viremic slow progression, and elite control. Efforts to identify differences between B57-positive (B57(+)) slow progressors and B57(+) rapid progressors have largely focused on cytotoxic T lymphocyte (CTL) phenotypes and specificities during chronic stages of infection. Although CTL responses in the early months of infection are likely to be the most important for the long-term rate of HIV-1 disease progression, few data on the early CTL responses of eventual slow progressors have been available. Utilizing the Multicenter AIDS Cohort Study (MACS), we retrospectively examined the early HIV-1-specific CTL responses of 14 B57(+) individuals whose time to development of disease ranged from 3.5 years to longer than 25 years after infection. In general, a greater breadth of targeting of epitopes from structural proteins, especially Gag, as well as of highly conserved epitopes from any HIV-1 protein, correlated with longer times until disease. The single elite controller in the cohort was an outlier on several correlations of CTL targeting and time until disease, consistent with reports that elite control is typically not achieved solely by protective HLA-mediated CTLs. When targeting of individual epitopes was analyzed, we found that early CTL responses to the IW9 (ISPRTLNAW) epitope of Gag, while generally subdominant, correlated with delayed progression to disease. This is the first study to identify early CTL responses to IW9 as a correlate of protection in persons with HLA-B*57.


Assuntos
Linfócitos T CD8-Positivos/imunologia , Regulação Viral da Expressão Gênica , HIV-1/metabolismo , Antígenos HLA-B/genética , Síndrome da Imunodeficiência Adquirida/sangue , Linfócitos T CD8-Positivos/virologia , Estudos de Coortes , Progressão da Doença , Epitopos/química , Humanos , Masculino , Modelos Estatísticos , Peptídeos/química , Fenótipo , Linfócitos T Citotóxicos/citologia , Produtos do Gene gag do Vírus da Imunodeficiência Humana/metabolismo
20.
J Virol ; 85(3): 1403-7, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21084486

RESUMO

We report the second human immunodeficiency virus (HIV) belonging to the new HIV type 1 (HIV-1) group P lineage that is closely related to the simian immunodeficiency virus found in gorillas. This virus was identified in an HIV-seropositive male hospital patient in Cameroon, confirming that the group P virus is circulating in humans. Results from screening 1,736 HIV-seropositive specimens collected in Cameroon indicate that HIV-1 group P infections are rare, accounting for only 0.06% of HIV infections. Despite its rarity, group P shows evidence of adaptation to humans.


Assuntos
Infecções por HIV/virologia , HIV-1/classificação , HIV-1/isolamento & purificação , Camarões , Genótipo , HIV-1/genética , Humanos , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Prevalência , Análise de Sequência de DNA , Vírus da Imunodeficiência Símia/genética
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