Decreased ability of HIV-1 Tat protein-treated accessory cells to organize cellular clusters is associated with partial activation of T cells

It has been shown in several animal models that HIV infection of accessory cells (ACs) plays an important role in development of AIDS. Here, we report that ACs treated with HIV-1 Tat protein (Tat-ACs) have a decreased ability to organize cellular aggregates as compared with untreated ACs, resulting in incomplete activation of T cells in responses to anti-CD3 mAb or staphylococcal enterotoxin B stimulation. The T cells failed to up-regulate adhesion molecules CD11a and CD2 on the cell surface and had reduced proliferative responses, as determined by [3H]thymidine incorporation, but they obtained lymphoblast-like morphology and expressed early activation antigens on the cell surface such as Fas and CD69 and interleukin 2 receptor, at comparable levels as those T cells undergoing a maximal proliferation. These results suggest that the Tat-AC-induced defect occurs in the late, but not in the early, phases of T cell activation. Normal expression of cell surface Fas antigen accompanied by defects in late activation thus may result in the susceptibility of these T cells to apoptosis. Our studies suggest that dysfunction, hyperactivation, and susceptibility to apoptosis, as observed with T cells isolated from HIV-infected individuals, may be, at least in part, a consequence of abnormal functions of ACs.

Tsg101, a homologue of ubiquitin-conjugating (E2) enzymes, binds the L domain in HIV type 1 Pr55Gag

Ubiquitination appears to be involved in virus particle release from infected cells. Free ubiquitin (Ub), as well as Ub covalently bound to a small fraction of p6 Gag, is detected in mature HIV particles. Here we report that the p6 region in the Pr55Gag structural precursor polyprotein binds to Tsg101, a putative Ub regulator that is involved in trafficking of plasma membrane-associated proteins. Tsg101 was found to interact with Gag in (i) a yeast two-hybrid assay, (ii) in vitro coimmunoprecipitation by using purified Pr55Gag and rabbit reticulocyte lysate-synthesized Tsg101, and (iii) in vivo in the cytoplasm of COS cells transfected with gag. The PTAPP motif [or late (L) domain] within p6, which is required for release of mature virus from the plasma membrane, was the determinant for binding Pr55Gag. The N-terminal region in Tsg101, which is homologous to the Ubc4 class of Ub-conjugating (E2) enzymes, was the determinant of interaction with p6. Mutation of Tyr-110 in Tsg101, present in place of the active-site Cys that binds Ub in E2 enzymes, and other residues unique to Tsg101, impaired p6 interaction, indicating that features that distinguish Tsg101 from active E2 enzymes were important for binding the viral protein. The results link L-domain function in HIV to the Ub machinery and a specific component of the cellular trafficking apparatus.

Selective pressure of a quinoxaline nonnucleoside inhibitor of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase (RT) on HIV-1 replication results in the emergence of nucleoside RT-inhibitor-specific (RT Leu-74-->Val or Ile and Val-75-->Leu or Ile) HIV-1 mutants.

The quinoxaline nonnucleoside RT inhibitor (NNRTI) (S)-4-isopropoxycarbonyl-6-methoxy-3-(methylthiomethyl)-3,4- dihydroquinoxaline-2(1H)-thione (HBY 097) was used to select for drug-resistant HIV-1 variants in vitro. The viruses first developed mutations affecting the NNRTI-binding pocket, and five of six strains displayed the RT G190-->E substitution, which is characteristic for HIV-1 resistance against quinoxalines. In one variant, a new mutant (G190-->Q) most likely evolved from preexisting G190-->E mutants. The negative charge introduced by the G190-->E substitution was maintained at that site of the pocket by simultaneous selection for V179-->D together with G190-->Q. After continued exposure to the drug, mutations at positions so far known to be specific for resistance against nucleoside RT inhibitors (NRTIs) (L74-->V/I and V75-->L/I) were consistently detected in all cultures. The inhibitory activities of the cellular conversion product of 2',3'-dideoxyinosine (ddI, didanosine), 2',3'-dideoxyadenosine (ddA) and of 2',3'-didehydro-3'-deoxythymidine (d4T, stavudine) against these late-passage viruses were shown to be enhanced with the L74-->V/I RT mutant virus as compared with the wild-type (wt) HIV-1MN isolate. Clonal analysis proved linkage of the codon 74 and codon 75 mutations to the NNRTI-specific mutations in all RT gene fragments. The nonnucleoside- and nucleoside-resistance mutation sites are separated by approximately 35 A. We propose that the two sites "communicate" through the template-primer which is situated in the DNA-binding cleft between these two sites. Quinoxalines cause high selective pressure on HIV-1 replication in vitro; however, the implication of these findings for the treatment of HIV-1 infection has yet to be determined.

Risco cardiovascular e sua associação com variáveis demográficas, clínicas e psicossociais em pessoas vivendo com HIV/aids

Trata-se de um estudo analítico de corte transversal, que visa avaliar o risco cardiovascular de PVHA segundo o Escore de Framingham e identificar a associação entre o risco e as variáveis demográficas, comportamentais, psicossociais e clínicas de PVHA. O estudo foi aprovado na Secretaria Municipal de Saúde e no Comitê de Ética da Escola de Enfermagem de Ribeirão Preto, a coleta de dados foi realizada no período de outubro de 2014 a agosto de 2015 em cinco Serviços de Atendimento Especializado às PVHA utilizando questionário sociodemográfico, clínico e comportamental, avaliação da alimentação saudável, Inventário de Sintomas de Stress para Adultos de Lipp e avaliação do risco cardiovascular por meio do Escore de Framingham. A análise dos dados ocorreu através de estatística descritiva e teste de associação entre as variáveis, onde foi adotado nível de significância com valor de p<0,05. Identificou-se que 58,3% pertenciam ao sexo masculino, 69,1% apresentavam idade acima de 40 anos, com média de 44,4 anos, 40,6% referiram ser brancos e 40,0% pardos, e 70,9% eram heterossexuais. Observou-se que 64,0% eram sedentários, 35,4% tabagistas e 40,0% faziam uso de bebida alcóolica regularmente. Do mesmo modo, 73,7% consideraram sua alimentação saudável, no entanto, ao ser avaliado de acordo com o escore da alimentação saudável, 70,9% obtiveram score intermediário para alimentação. Com relação às variáveis psicossociais, foi identificado que 52,0% tinham menos de oito anos de estudo, e 80,6% referiram receber até três salários mínimos por mês. Quanto aos sintomas de estresse, foi visto que 29,1% e 22,3% estavam nas fases de resistência e exaustão, respectivamente. Além disso, identificou-se que 15,4% da amostra tinha diagnóstico médico para depressão e que 71,4% não realizavam atividades de lazer regularmente. Com relação às variáveis clínicas gerais, 57,7% referiram antecedentes familiares para HAS, 40,6% para DM, 21,7% para IAM e 27,4% para AVE. Quanto aos antecedentes pessoais, foi visto que 15,4% eram hipertensos, 8,0% eram diabéticos e 8,0% tinham dislipidemia. Desta mesma amostra, 45,2% apresentavam IMC maior que 25,0 kg/m² e 41,7% estavam em síndrome metabólica. Com relação às variáveis clínicas relacionadas ao HIV, observou-se que 42,2% e 32,0% possuíam o diagnóstico de soropositividade e fazem uso de TARV há mais de dez anos, respectivamente. A contagem de células TCD4+ e carga viral mostrou que 82,8% dos participantes apresentaram contagem maior que 350 cels/mm³, e 80,6% tinham carga viral indetectável. Foi identificado que 25,8% dos sujeitos apresentam risco cardiovascular de médio a alto, segundo o Escore de Framingham. Apenas as variáveis sociodemográficas sexo (p=0,006), idade (p<0,001) e estado civil (p=0,003) apresentaram associação com o risco cardiovascular calculado pelo Escore de Framingham. Nas variáveis comportamentais, as fases de estresse (p=0,039) tiveram associação com o risco cardiovascular, e com relação às variáveis clínicas, antecedentes familiares para DM (p=0,035), HAS, DM e SM (p<0,001) e DLP (p=0,030) apresentaram significância estatística. Nas variáveis clínicas relacionadas ao HIV, o tempo de diagnóstico (p=0,005) e o tempo de TARV (p=0,038) também apresentaram associação. Conclui-se que 25,8% de PVHA no município de Ribeirão Preto apresentam risco cardiovascular de moderado a alto, medido pelo Escore de Framingham

Immune recovery in HIV-infected patients after candida esophagitis is impaired despite long-term antiretroviral therapy.

OBJECTIVE Candida esophagitis belongs to the most common AIDS-defining diseases, however, a comprehensive immune pathogenic concept is lacking. DESIGN We investigated the immune status of 37 HIV-1-infected patients from the Swiss HIV cohort study at diagnosis of Candida esophagitis, 1 year before, 1 year later and after 2 years of suppressed HIV RNA. We compared these patients to 3 groups: 37 HIV-1-infected patients without Candida esophagitis but similar CD4 counts as the patients at diagnosis (advanced HIV group), 15 HIV-1-infected patients with CD4 counts >500 cells/μl, CD4 nadir >350 cells/μl and suppressed HIV RNA under combination antiretroviral therapy (cART) (early cART group), and 20 healthy individuals. METHODS We investigated phenotype, cytokine production and proliferative capacity of different immune cells by flow cytometry and ELISpot. RESULTS We found that patients with Candida esophagitis had nearly abolished CD4 proliferation in response to C. albicans, significantly increased percentages of dysfunctional CD4 cells, significantly decreased cytotoxic NK-cell counts and peripheral innate lymphoid cells and significantly reduced IFN-γ and IL-17 production compared to the early cART group and healthy individuals. Most of these defects remained for more than 2 years despite viral suppression. The advanced HIV group without opportunistic infection showed partly improved immune recovery. CONCLUSIONS Our data indicate that Candida esophagitis in HIV-1-infected patients is caused by an accumulation of multiple, partly Candida-specific immunological defects. Long-term immune recovery is impaired, illustrating that specific immunological gaps persist despite cART. These data also support the rationale for early cART initiation to prevent irreversible immune defects.

Reading and interpreting the HIV-1 Western Blot test /

Accompanying material in pockets, front and back covers.

HIV Status Disclosure among Older African American Men Who Have Sex with Men and Women

Thesis (Ph.D.)--University of Washington, 2016-06

Diplacanthid acanthodians from the Aztec Siltstone (late Middle Devonian) of southern Victoria Land, Antartica

One articulated and several partial, semi-articulated specimens of acanthodians were collected in 1970 from the freshwater deposits of the Aztec Siltstone (Middle Devonian; Givetian), Portal Mountain, southern Victoria Land, Antarctica, during a Victoria University of Wellington Antarctic Expedition. The Portal Mountain fish fauna, preserved in a finely laminated, non-calcareous siltstone, includes acanthodians, palaeoniscoids, and bothriolepid placoderms. The articulated acanthodian specimens are the most complete fossil fish remains documented so far from the Aztec assemblage, which is the most diverse fossil vertebrate fauna known from Antarctica. They are described as a new taxon, Milesacanthus antarctica gen. et sp. nov., which is assigned to the family Diplacanthidae. Its fin spines show some similarities to spine fragments named Byssacanthoides debenhami from glacial moraine at Granite Harbour, Antarctica, and much larger spines named Antarctonchus glacialis from outcrops of the Aztec Siltstone in the Boomerang Range, southern Victoria Land. Both of these are reviewed, and retained as form taxa for isolated spines. Various isolated remains of fin spines and scales are described from Portal Mountain and Mount Crean (Lashly Range), and referred to Milesacanthus antarctica gen. et sp. nov. The histology of spines and scales is documented for the first time, and compared with acanthodian material from the Devonian of Australia and Europe. Distinctive fin spines from Mount Crean are provisionally assigned to Culmacanthus antarctica Young, 1989b. Several features on the most complete of the new fish specimens - in particular, the apparent lack of an enlarged cheek plate - suggest a revision of the diagnosis for the Diplacanthidae.

Symptoms and diagnosis of borderline, early and advanced epithelial ovarian cancer

Objective. To document symptoms associated with borderline, early and advanced ovarian cancer and identify personal characteristics associated with early versus late diagnosis. Methods. Information concerning symptoms and diagnosis history was available from 811 women with ovarian cancer who took part in an Australian case–control study in the early 1990s. Women were classified into three groups for comparison based on their diagnosis: borderline, early (stage I–II) and advanced (stage III–IV) invasive cancer. Results. Sixteen percent of women with borderline tumors, 7% with early cancer and 4% with advanced cancer experienced no symptoms before diagnosis (P < 0.0001). Among women with symptoms, abdominal pain (44%) or swelling (39%) were most frequently reported; an abdominal mass (12%) and gynecological symptoms (12%) were less common. Compared to advanced stage cancer, women with early stage cancer were more likely to report an abdominal mass or urinary symptoms but less likely to report gastrointestinal problems or general malaise. General malaise and ‘other’ symptoms were least common in borderline disease. Older women, and those with higher parity or a family history of breast or ovarian cancer, were more likely to be diagnosed at an advanced stage of disease. Conclusions. Women who experience persistent or recurrent abdominal symptoms, particularly swelling and/or pain should be encouraged to seek medical attention and physicians should be alert to the possibility of ovarian cancer even in the absence of an abdominal mass. Further information about the prevalence of these symptoms in the general population is essential to assist physicians in patient management.

Population pharmacokinetics of metformin in late pregnancy

The pharmacokinetic disposition of metformin in late pregnancy was studied together with the level of fetal exposure at birth. Blood samples were obtained in the third trimester of pregnancy from women with gestational diabetes or type 2 diabetes, 5 had a previous diagnosis of polycystic ovary syndrome. A cord blood sample also was obtained at the delivery of some of these women, and also at delivery of others who had been taking metformin during pregnancy but from whom no blood had been taken. Plasma metformin concentrations were assayed by a new, validated, reverse-phase HPLC method, A 2-compartment, extravascular maternal model with transplacental partitioning of drug to a fetal compartment was fitted to the data. Nonlinear mixed-effects modeling was performed in'NONMEM using FOCE with INTERACTION. Variability was estimated using logarithmic interindividual and additive residual variance models; the covariance between clearance and volume was modeled simultaneously. Mean (range) metformin concentrations in cord plasma and in maternal plasma were 0.81 (range, 0.1-2.6) mg/L and 1.2 (range, 0. 1-2.9) mg/L, respectively. Typical population values (interindividual variability, CV%) for allometrically scaled maternal clearance and volume of distribution were 28 L/h/70 kg (17.1%) and 190 L/70 ka (46.3%), giving a derived population-wide half-life of 5.1 hours. The placental partition coefficient for metformin was 1.07 (36.3%). Neither maternal age nor weight significantly influenced the pharmacokinetics. The variability (SD) of observed concentrations about model-predicted concentrations was 0.32 mg/L. The pharmacokinetics were similar to those in nonpregnant patients and, therefore, no dosage adjustment is warranted. Metformin readily crosses the placenta, exposing the fetus to concentrations approaching those in the maternal circulation. The sequelae to such exposure, ea, effects on neonatal obesity and insulin resistance, remain unknown.

theta-defensins prevent HIV-1 Env-mediated fusion by binding gp41 and blocking 6-helix bundle formation

Retrocyclin-1, a 0-defensin, protects target cells from human immunodeficiency virus, type 1 (HIV-1) by preventing viral entry. To delineate its mechanism, we conducted fusion assays between susceptible target cells and effector cells that expressed HIV-1 Env. Retrocyclin-1 (4 mu M) completely blocked fusion mediated by HIV-1 Envs that used CXCR4 or CCR5 but had little effect on cell fusion mediated by HIV-2 and simian immunodeficiency virus Envs. Retrocyclin-1 inhibited HIV-1 Env-mediated fusion without impairing the lateral mobility of CD4, and it inhibited the fusion of CD4-deficient cells with cells bearing CD4-independent HIV-1 Env. Thus, it could act without cross-linking membrane proteins or inhibiting gp120-CD4 interactions. Retrocyclin-1 acted late in the HIV-1 Env fusion cascade but prior to 6-helix bundle formation. Surface plasmon resonance experiments revealed that retrocyclin bound the ectodomain of gp41 with high affinity in a glycan-independent manner and that it bound selectively to the gp41 C-terminal heptad repeat. Native-PAGE, enzyme-linked immunosorbent assay, and CD spectroscopic analyses all revealed that retrocyclin-1 prevented 6-helix bundle formation. This mode of action, although novel for an innate effector molecule, resembles the mechanism of peptidic entry inhibitors based on portions of the gp41 sequence.

Prevalence of drug-resistance mutations and non-subtype B strains among HIV-infected infants from New York State

Summary: Prevalence studies indicate that transmission of drug-resistant HIV has been rising in the adult population, but data from the perinatally infected pediatric population are limited. In this retrospective study, we sequenced the pol region of HIV from perinatally infected infants diagnosed in New York State in 2001-2002. Analyses of drug resistance, subtype diversity, and perinatal antiretroviral exposure were conducted, and the results were compared with those from a previous study of HIV-infected infants identified in 1998-1999. Eight of 42 infants (19.1%) had provirus carrying at least 1 drug-resistance mutation, an increase of 58% over the 1998-1999 results. Mutations conferring resistance to nucleoside reverse transcriptase inhibitors, nonnucleoside reverse transcriptase inhibitors, and protease inhibitors were detected in 7.1%, 11.9%, and 2.4% of specimens, respectively. Consistent with previous results, perinatal antiretroviral exposure was not associated with drug resistance (P = 0.70). Phylogenetic analysis indicated that 16.7% of infants were infected with a non-subtype B strain of HIV. It seems that drug-resistant and non-subtype B strains of HIV are becoming increasingly common in the perinatally infected population. Our results highlight the value of resistance testing for all HIV-infected infants upon diagnosis and the need to consider subtype diversity in diagnostic and treatment strategies.

EFICÁCIA ADAPTATIVA E SITUAÇÕES DE CRISE EM PORTADORES DO VÍRUS HIV IMUNODEFICIÊNCIA HUMANA / AIDS.

O objetivo desta pesquisa foi investigar a eficácia adaptativa e situações de crise de indivíduos portadores do vírus HIV do Programa Municipal DST/AIDS de Aparecida SP. O instrumento utilizado foi a Entrevista Clínica Preventiva - EDAO (Escala Diagnóstica Adaptativa Operacionalizada). Participaram do estudo 5 homens e 5 mulheres que freqüentavam o serviço de saúde. Os resultados do trabalho revelaram que ser portador de uma doença crônica carregada de estigmas como a AIDS é um fator desestruturante para o diagnóstico. Foi percebida na população estudada o comprometimento dos setores afetivo-relacional e produtividade, seguido do sóciocultural. Foram observados alguns aspectos importantes como: a crise do impacto diagnóstico como sendo algo marcante em todos os participantes, bem como o uso de drogas, comportamentos vulneráveis que possivelmente levaram à infecção pelo vírus HIV, dificuldades de resolução de conflitos e nas relações interpessoais, perdas vivenciadas durante toda a vida e também no decorrer da infecção e o fenômeno da feminização do vírus HIV através das mulheres entrevistadas, que foram infectadas por seus parceiros sexuais estáveis. No setor orgânico de funcionamento, a população estudada mostrou adesão ao tratamento A maioria da população estudada foi diagnosticada com adaptação ineficaz severa. Este estudo trouxe questionamentos importantes sobre a maneira com a qual o indivíduo portador do vírus HIV mantém seu equilíbrio psíquico e suas relações com o trabalho que executa, chamando atenção para a necessidade de outros estudos que contemplem diferentes instrumentos para a compreensão do tema.

ASPECTOS PSICOLÓGICOS DE PACIENTES COM HIV/AIDS E LIPODISTROFIA ATENDIDOS NO HOSPITAL HELIÓPOLIS

A epidemia da Síndrome da Imunodeficiência Adquirida (aids) é, atualmente, um fenômeno de grande magnitude e extensão na saúde mundial. A síndrome de lipodistrofia é uma alteração que afeta a autoimagem corporal e a sexualidade, aumentando o estigma da doença e ocasionando dificuldades na adesão ao tratamento e nas relações sociais. OBJETIVOS: a) descrever aspectos da psicodinâmica de pacientes HIV/aids acometidos e não acometidos pela lipodistrofia, dando enfoque aos mecanismos de defesa utilizados ; b) investigar a percepção de imagem corporal em pacientes HIV/aids acometidos e não acometidos pela síndrome de lipodistrofia; c) identificar semelhanças e diferenças de percepção de imagem corporal em pacientes HIV/aids acometidos pela lipodistrofia com aqueles não acometidos. MÉTODO: Foram selecionados oito pacientes por critério de conveniência do ambulatório da Clínica de Infectologia do Hospital Heliópolis. Foram utilizados um Roteiro de Entrevista e o Desenho da Figura Humana DFH teste projetivo gráfico de personalidade; a análise dos dados foram submetidos à análise qualitativa conforme indicação do instrumental, auxiliados pela leitura do conteúdo clinico-diagnóstico psicológico. RESULTADOS: Nos dois grupos os dados apontaram para características em comum quanto à psicodinâmica interna e à percepção de imagem corporal. Recursos defensivos primitivos foram os mais utilizados caracterizando a presença de disfunção da imagem corporal e um controle egóico rígido, embora frágil. Percebeu-se o quanto é angustiante, para estas pacientes, lidar não somente com a autoimagem como também com a sexualidade. CONCLUSÕES: Os programas de acompanhamento ao HIV/aids devem considerar o quanto essas pacientes necessitam de ser acompanhadas em psicoterapia. A promoção de saúde deve levar em conta não somente a melhora da qualidade de vida, mas também buscar compreender como estas mulheres se relacionam e de que forma exercem a sua sexualidade.

The spatial patterns of β-amyloid (Aβ) deposits and neurofibrillary tangles (NFT) in late-onset sporadic Alzheimer's disease

The spatial patterns of β-amyloid (Aβ) deposits and neurofibrillary tangles (NFT) were studied in areas of the cerebral cortex in 16 patients with the late-onset, sporadic form of Alzheimer's disease (AD). Diffuse, primitive, and classic Aβ deposits and NFT were aggregated into clusters; the clusters being regularly distributed parallel to the pia mater in many areas. In a significant proportion of regions, the sizes of the regularly distributed clusters approximated to those of the cells of origin of the cortico-cortical projections. The diffuse and primitive Aβ deposits exhibited a similar range of spatial patterns but the classic Aβ deposits occurred less frequently in large clusters >6400m. In addition, the NFT often occurred in larger regularly distributed clusters than the Aβ deposits. The location, size, and distribution of the clusters of Aβ deposits and NFT supports the hypothesis that AD is a 'disconnection syndrome' in which degeneration of specific cortico-cortical and cortico-hippocampal pathways results in synaptic disconnection and the formation of clusters of NFT and Aβ deposits. © 2011 Nova Science Publishers, Inc.