Frequency of subtype B and F1 dual infection in HIV-1 positive, Brazilian men who have sex with men

Introdução: Como varios estudos sobre vacinacao contra HIV estao em progresso, e importante compreender a frequencia na qual ocorrem co/superinfeccoes intra ou intersubtipos em grupos de alto risco. Esse conhecimento auxiliaria no desenvolvimento de programas de prevencao futuros. Nesse estudo transversal, relatamos a frequencia de coinfeccao entre os subtipos B e F1 em uma cohorte clinica de 41 homens que fazem sexo com homens (HSH), recem infectados com HIV-1, em São Paulo, Brasil. Metodologia: O DNA proviral do HIV-1 foi isolado a partir de leucocitos de sangue periferico de sujeitos polimorfonucleares (PMNs), que foram obtidos no momento da inscricao. Cada individuo era conhecidamente infectado por um virus do subtipo B, conforme determinado em estudo anterior. Um pequeno fragmento do gene da integrase (nucleotideo 4255-4478 do HXB2) foi amplificado por PCR utilizando primers especificos para F1. Os resultados da PCR foram confirmados por analise filogenetica. Os dados de carga viral (VL) foram inferidos a partir dos prontuarios de cada paciente. Resultados: Das 41 amostras estudadas, 5 apresentaram DNA proviral do subtipo F1, O que representa uma taxa de 12,2% de coinfeccao. A comparacao entre os valores de carga viral entre os coinfectados e os infectados apenas pelo subtipo B nao foi estatisticamente diferente (p> 0,16). Nos individuos com infeccao dupla a carga viral mediana foi de 5,3 x 104 copias/mL (intervalo de <400 u 12,5 x 104 copias/mL), e nos individuos infectados apenas pelo subtipo B a carga viral mediana foi de 4,3 x 104 copias/mL (intervalo de <400 u 39,9 x 104 copias/mL). Conclusao: Esse estudo indicou que a coinfeccao entre os subtipos B e F1 ocorre com frequencia na populacao de homens que fazem sexo com homens, HIV-1 positivos, como sugerido por um grande numero de virus recombinantes BF1 relatados no Brasil. Na ausencia de uma vacina eficaz contra o HIV-1, o teste para co/superinfeccao e a implantacao de medidas eficazes nos grupos de risco podem ajudar a reduzir a exposicao viral, a transmissao e a recombinacaoBV UNIFESP: Teses e dissertaçõe

Variability of HIV-1 Genomes among Children and Adolescents from São Paulo, Brazil

Background: Genetic variability is a major feature of the human immunodeficiency virus type 1 (HIV-1) and considered the key factor to frustrating efforts to halt the virus epidemic. in this study, we aimed to investigate the genetic variability of HIV-1 strains among children and adolescents born from 1992 to 2009 in the state of São Paulo, Brazil.Methodology: Plasma and peripheral blood mononuclear cells (PBMC) were collected from 51 HIV-1-positive children and adolescents on ART followed between September 1992 and July 2009. After extraction, the genetic materials were used in a polymerase chain reaction (PCR) to amplify the viral near full length genomes (NFLGs) from 5 overlapped fragments. NFLGs and partial amplicons were directly sequenced and data were phylogenetically inferred.Results: of the 51 samples studied, the NFLGs and partial fragments of HIV-1 from 42 PBMCs and 25 plasma were successfully subtyped. Results based on proviral DNA revealed that 22 (52.4%) patients were infected with subtype B, 16 (38.1%) were infected with BF1 mosaic variants and 4 (9.5%) were infected with sub-subtype F1. All the BF1 recombinants were unique and distinct from any previously identified unique or circulating recombinant forms in South America. Evidence of dual infections was detected in 3 patients coinfected with the same or distinct HIV-1 subtypes. Ten of the 31 (32.2%) and 12 of the 21 (57.1%) subjects with recovered proviral and plasma, respectively, protease sequences were infected with major mutants resistant to protease inhibitors. the V3 sequences of 14 patients with available sequences from PBMC/or plasma were predicted to be R5-tropic virus except for two patients who harbored an X4 strain.Conclusions: the high proportion of HIV-1 BF1 recombinant, coinfection rate and vertical transmission in Brazil merits urgent attention and effective measures to reduce the transmission of HIV among spouses and sex partners.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)National Institutes of HealthUniv São Paulo, Sch Med, Clin & Res Lab LIM 03, São Paulo, BrazilUniv São Paulo, Inst Trop Med, Virol Lab LIM HCFMUSP 52, São Paulo, BrazilUniv Florida, Miller Sch Med, Story Lab 2, Miami, FL USAUniversidade Federal de São Paulo, Paulista Sch Med, Dept Pediat, São Paulo, BrazilUniv São Paulo, Sch Med, Div Clin Immunol & Allergy, São Paulo, BrazilUniv Calif San Francisco, Dept Med, Div Expt Med, San Francisco, CA USAUniversidade Federal de São Paulo, Paulista Sch Med, Dept Pediat, São Paulo, BrazilFAPESP: 2011/09983-1FAPESP: 2009/540055-5FAPESP: 2009/52381-2FAPESP: 2010/05845-0 2004/15856-9FAPESP: 2006/50096-0CAPES: 2571/2009National Institutes of Health: R01 AI060379Web of Scienc

Luminescence investigation of the Sm(III)-b-diketonates with sulfoxides, phosphine oxides and amides ligands.

In this paper we report on a photoluminescent investigation of complexes involving Sm -b-diketonates with sulfoxides, phosphine oxides and amides ligands. In the synthesis of the coordination compounds we used samarium tris(thenoyltrifluoroacetonate) dehydrated precursor with the following ligands (L): DBSO and PTSO sulfoxides; TPPO phosphine oxide and (PHA) N-phenylacetamide. They have 31 31 shown high orange luminescence characteristic of the Sm ion. The emission spectra of the Sm -complexes present narrow bands 4 6 4 6 arising from the G →H (J55/ 2, 7/ 2, 9/ 2, 11/2) transitions with the hypersensitive G →H transition as a prominent group. It 5 / 2 J 5 / 2 9/ 2 4 31 is observed an efficient intramolecular energy transfer from the triplet state (T) of the ligands to the emitting G state of the Sm ion. 5 / 2 The experimental intensity parameters (h andh ) for the Sm and Eu complexes have been determined and compared. The lifetimes (t ) Sm Eu 4 of the emitting level G of the Sm-complexes are approximately 10 times higher than in the precursor compound [Sm(TTA) ?(H O) ] 5 / 2 3 2 2 indicating that radiative processes are operative in all the compounds due to the absence of multiphonon relaxation by coupling with the OH oscillators

Genetic distances and phylogenetic tree constructed using a maximum-likelihood method of HIV isolates recovered from patient 01BR_IMT_041.

<p>A: Phylogenetic tree from concatenated regions assigned as subtype B from the BF1 recombinant isolate. B: Phylogenetic tree showing the clustering pattern of F1 sequences (marked by dotted box). F1 region from genuine F1 sequence recovered from plasma and PBMCs are marked by a black circle and square, respectively while the F1 region from the BF1 recombinant sequence is marked by an empty square. The approximate likelihood ratio test (aLRT) values of ≥70% are indicated at nodes. The scale bar represents 0.05 nucleotide substitutions per site.</p

Drug-resistance mutations detected in PBMC.

1<p>Displayed insertion at position 69.</p><p>Regions not sequenced are indicated by empty boxes.</p><p>High resistance mutations are indicated by bold lettering.</p

Schematic representation of the NFLG, partial structure and breakpoint profiles of the BF1 sequences identified in this study both from HIV RNA and proviral DNA.

<p>Samples that were identified in this study to host distinct viruses are indicated with the star symbol. The region of subclade F1 and subtypes B are indicated at the bottom.</p

Patient characteristics for the study population.

<p>Patient characteristics for the study population.</p

Genetic distances and phylogenetic tree constructed using a maximum-likelihood method from concatenated regions of HIV RNA (indicated by black circles) and proviral DNA (indicated by black squares) marked with Arabic numbers 1 and 2 from patient 010BR_IMT_020 along with HIV-1 reference sequences from the Los Alamos HIV-1 database representing 11 genetic subtypes.

<p>For purposes of clarity, the tree was midpoint rooted. The approximate likelihood ratio test (aLRT) values of ≥70% are indicated at nodes. The scale bar represents 0.05 nucleotide substitutions per site.</p

Maximum-likelihood phylogenetic trees form each non-recombinant fragment were constructed using all available sequences from proviral DNA (indicated by black circles) and plasma isolate (indicated by an empty circles) along with HIV-1 reference sequences from the Los Alamos HIV-1 database representing 11 genetic subtypes.

<p>The numbering for the HIV-1 fragment A, B1, B2 and C sequences corresponds to the HXB2 reference sequence. For purposes of clarity, the tree was midpoint rooted. The approximate likelihood ratio test (aLRT) values of ≥70% are indicated at nodes. The scale bar represents 0.05 nucleotide substitutions per site.</p