Predominance of CXCR4 tropism in HIV-1 CRF14_BG strains from newly diagnosed infections.

OBJECTIVES: R5-tropic viruses are associated with HIV-1 transmission and predominate during the early stages of infection. X4-tropic populations have been detected in ~50% of patients with late-stage disease infected with subtype B viruses. In this study, we compared the frequency of X4 tropism in individuals infected with HIV-1 CRF14_BG viruses, which have a V3 loop of subtype B, with a control group of individuals infected with subtype B viruses. METHODS: Sixty-three individuals infected with HIV-1 CRF14_BG (n = 31) or subtype B (n = 32) were studied. Similar proportions of newly diagnosed and chronically infected individuals were included in the subtype B and CRF14_BG groups. V3 sequences were obtained and coreceptor tropism was predicted using the Geno2pheno[coreceptor] algorithm. V3 net charge and 11/25 rules were also used for coreceptor prediction. RESULTS: Overall, X4 tropism was more frequent among individuals infected with CRF14_BG viruses (87.1%) than subtype B viruses (34.3%), a difference that was statistically highly significant (P = 0.00001). Importantly, the frequencies among newly diagnosed individuals were 90% and 13.3%, respectively (P = 0.0007). Characteristic amino acids in the V3 loop (T13, M14, V19 and W20) were identified at higher frequencies in CRF14_BG viruses (54%) than subtype B viruses (0%; P < 0.000001). CONCLUSIONS: CRF14_BG is the genetic form with the highest proportion of X4-tropic viruses reported to date in newly diagnosed and chronic infections. This suggests high pathogenicity for CRF14_BG viruses, potentially leading to rapid disease progression. CCR5 antagonists will be ineffective in most CRF14_BG-infected patients, even at early stages of infection.

Improvement of HIV-1 coreceptor tropism prediction by employing selected nucleotide positions of the env gene in a Bayesian network classifier.

OBJECTIVES: This study aimed to develop a genotypic method to predict HIV-1 coreceptor usage by employing the nucleotide sequence of the env gene in a tree-augmented naive Bayes (TAN) classifier, and to evaluate its accuracy in prediction compared with other available tools. METHODS: A wrapper data-mining strategy interleaved with a TAN algorithm was employed to evaluate the predictor value of every single-nucleotide position throughout the HIV-1 env gene. Based on these results, different nucleotide positions were selected to develop a TAN classifier, which was employed to predict the coreceptor tropism of all the full-length env gene sequences with information on coreceptor tropism currently available at the Los Alamos HIV Sequence Database. RESULTS: Employing 26 nucleotide positions in the TAN classifier, an accuracy of 95.6%, a specificity (identification of CCR5-tropic viruses) of 99.4% and a sensitivity (identification of CXCR4/dual-tropic viruses) of 80.5% were achieved for the in silico cross-validation. Compared with the phenotypic determination of coreceptor usage, the TAN algorithm achieved more accurate predictions than WebPSSM and Geno2pheno [coreceptor] (P<0.05). CONCLUSIONS: The use of the methodology presented in this work constitutes a robust strategy to identify genetic patterns throughout the HIV-1 env gene differently present in CCR5-tropic and CXCR4/dual-tropic viruses. Moreover, the TAN classifier can be used as a genotypic tool to predict the coreceptor usage of HIV-1 isolates reaching more accurate predictions than with other widely used genotypic tools. The use of this algorithm could improve the correct prescribing of CCR5 antagonist drugs to HIV-1-infected patients.

Evaluation of genotypic tropism prediction tests compared with in vitro co-receptor usage in HIV-1 primary isolates of diverse subtypes.

OBJECTIVES: To evaluate the sensitivity and specificity of genotypic methods for predicting the co-receptor usage of subtypes B and non-B HIV-1 primary isolates, using as gold standard the infectivity of each primary isolate in GHOST cells stably expressing HIV-1 co-receptors. METHODS: Primary isolates were obtained by co-culturing either patient's peripheral blood mononuclear cells (PBMCs) or ultracentrifuged plasma with donor-activated PBMCs. In vitro co-receptor usage was determined by infecting GHOST cells. Tropism prediction, based on V3 sequences, was determined with simple rules and bioinformatic tools (Geno2pheno[coreceptor] and WebPSSM). RESULTS: This study includes 102 HIV-1 primary isolates; 23 (22.5%) subtype B and 79 (77.5%) non-B genetic forms. V3 sequences were classified into six subtypes (A-G), although 32 (31.4%) were circulating recombinant forms and 21 (20.6%) were unique recombinant forms. Sixty-nine isolates were R5, 27 R5X4 and 6 X4. The highest levels of sensitivity and specificity for the detection of X4 strains among V3 sequences, between 91% and 100%, were obtained by using PSSM(x4r5), PSSM(si/nsi) and the 11/25 rule for sequences of subtypes A, B and G, but not for subtype F. Establishing the recommended cut-off for clinical settings of a 10% false positive rate for Geno2pheno, we obtained 93% specificity and 97% sensitivity. CONCLUSIONS: Comparing genotypic assays for HIV-1 co-receptor use with a cell-culture phenotypic assay could provide more reliable results of sensitivity and specificity for the detection of X4 strains than comparing them with recombinant assays, considered as gold standard. In general, except for subtype F isolates, there is a good correlation for tropism prediction.

New insights into the origin of the HIV type 1 subtype A epidemic in former Soviet Union's countries derived from sequence analyses of preepidemically transmitted viruses.

The HIV-1 subtype A epidemic affecting injecting drug users (IDU) in former Soviet Union (FSU) countries started dramatically in Odessa, southern Ukraine, in 1995, and is caused by a variant of monophyletic origin, often designated IDU-A. We phylogenetically analyzed one near full-length genome and two partial sequences of three HIV-1 subtype A viruses collected in St. Petersburg, Russia, heterosexually transmitted in 1992-1994. The sequences branched basally to the IDU-A clade, together with eight viruses from Odessa collected in 1993, all presumably acquired heterosexually, and two viruses from the Democratic Republic of Congo. Of all other FSU sequences in databases, only those from three recently collected viruses, one from Ukraine and two from northwestern Russia, at least one of them acquired heterosexually, branched basally to the IDU-A cluster. The results indicate that the FSU IDU-A variant derives from a strain that initially propagated heterosexually in Ukraine and originated in central Africa.

Identification of an HIV-1 BG Intersubtype Recombinant Form (CRF73_BG), Partially Related to CRF14_BG, Which Is Circulating in Portugal and Spain.

HIV-1 exhibits a characteristically high genetic diversity, with the M group, responsible for the pandemic, being classified into nine subtypes, 72 circulating recombinant forms (CRFs) and numerous unique recombinant forms (URFs). Here we characterize the near full-length genome sequence of an HIV-1 BG intersubtype recombinant virus (X3208) collected in Galicia (Northwest Spain) which exhibits a mosaic structure coincident with that of a previously characterized BG recombinant virus (9601_01), collected in Germany and epidemiologically linked to Portugal, and different from currently defined CRFs. Similar recombination patterns were found in partial genome sequences from three other BG recombinant viruses, one newly derived, from a virus collected in Spain, and two retrieved from databases, collected in France and Portugal, respectively. Breakpoint coincidence and clustering in phylogenetic trees of these epidemiologically-unlinked viruses allow to define a new HIV-1 CRF (CRF73_BG). CRF73_BG shares one breakpoint in the envelope with CRF14_BG, which circulates in Portugal and Spain, and groups with it in a subtype B envelope fragment, but the greatest part of its genome does not appear to derive from CRF14_BG, although both CRFs share as parental strain the subtype G variant circulating in the Iberian Peninsula. Phylogenetic clustering of partial pol and env segments from viruses collected in Portugal and Spain with X3208 and 9691_01 indicates that CRF73_BG is circulating in both countries, with proportions of around 2-3% Portuguese database HIV-1 isolates clustering with CRF73_BG. The fact that an HIV-1 recombinant virus characterized ten years ago as a URF has been shown to represent a CRF suggests that the number of HIV-1 CRFs may be much greater than currently known.

Sequence Analysis of In Vivo-Expressed HIV-1 Spliced RNAs Reveals the Usage of New and Unusual Splice Sites by Viruses of Different Subtypes.

HIV-1 RNAs are generated through a complex splicing mechanism, resulting in a great diversity of transcripts, which are classified in three major categories: unspliced, singly spliced (SS), and doubly spliced (DS). Knowledge on HIV-1 RNA splicing in vivo and by non-subtype B viruses is scarce. Here we analyze HIV-1 RNA splice site usage in CD4+CD25+ lymphocytes from HIV-1-infected individuals through pyrosequencing. HIV-1 DS and SS RNAs were amplified by RT-PCR in 19 and 12 samples, respectively. 13,108 sequences from HIV-1 spliced RNAs, derived from viruses of five subtypes (A, B, C, F, G), were identified. In four samples, three of non-B subtypes, five 3' splice sites (3'ss) mapping to unreported positions in the HIV-1 genome were identified. Two, designated A4i and A4j, were used in 22% and 25% of rev RNAs in two viruses of subtypes B and A, respectively. Given their close proximity (one or two nucleotides) to A4c and A4d, respectively, they could be viewed as variants of these sites. Three 3'ss, designated A7g, A7h, and A7i, located 20, 32, and 18 nucleotides downstream of A7, respectively, were identified in a subtype C (A7g, A7h) and a subtype G (A7i) viruses, each in around 2% of nef RNAs. The new splice sites or variants of splice sites were associated with the usual sequence features of 3'ss. Usage of unusual 3'ss A4d, A4e, A5a, A7a, and A7b was also detected. A4f, previously identified in two subtype C viruses, was preferentially used by rev RNAs of a subtype C virus. These results highlight the great diversity of in vivo splice site usage by HIV-1 RNAs. The fact that four of five newly identified splice sites or variants of splice sites were detected in non-subtype B viruses allows anticipating an even greater diversity of HIV-1 splice site usage than currently known.

The analysis of near full-length genome sequences of human immunodeficiency virus type 1 BF intersubtype recombinant viruses from Chile, Venezuela and Spain reveals their relationship to diverse lineages of recombinant viruses related to CRF12_BF.

Human immunodeficiency virus type 1 (HIV-1) BF intersubtype recombinant viruses are common in Argentina and Uruguay, where CRF12_BF and related recombinants are frequently found, and, in a lower proportion, in Brazil. Full-length genome sequences have been characterized in several of these recombinant viruses. Here, we analyze six newly derived near full-length genome sequences of BF recombinant viruses, three from Chile, one from Venezuela and two from Spain. Five of them had known epidemiological links to Argentina. Genomes were amplified by PCR from plasma RNA or from peripheral blood mononuclear cells' DNA. Mosaic structures and phylogenetic relationships were analyzed by bootscanning, neighbour-joining phylogenetic trees and by examination of subtype signature nucleotides. One virus from Spain had a mosaic structure fully coincident with CRF12_BF. The others had unique mosaic structures, except the viruses from two Chilean sisters infected vertically from the same mother, who showed identical recombination patterns. Each of the unique recombinants had one to six breakpoints coincident with CRF12_BF and three also had two or three breakpoints coincident with a previously characterized unique recombinant from Argentina (A025) related to CRF12_BF. A phylogenetic tree of concatenated subtype F segments supported the relationship of five recombinants with CRF12_BF. In trees of partial subtype F and B segments, four recombinants clustered with A025. The examination of CRF12_BF signature amino acids and nucleotides supported the common ancestry of all the analyzed viruses. Based on these results, a model of generation of HIV-1 BF recombinants of Argentinean ancestry by successive rounds of recombination along diverse lineages deriving from a common BF recombinant ancestor related to CRF12_BF is proposed.

Epidemiological Surveillance of HIV-1 Transmitted Drug Resistance in Spain in 2004-2012: Relevance of Transmission Clusters in the Propagation of Resistance Mutations.

Our objectives were to carry out an epidemiological surveillance study on transmitted drug resistance (TDR) among individuals newly diagnosed of HIV-1 infection during a nine year period in Spain and to assess the role of transmission clusters (TC) in the propagation of resistant strains. An overall of 1614 newly diagnosed individuals were included in the study from January 2004 through December 2012. Individuals come from two different Spanish regions: Galicia and the Basque Country. Resistance mutations to reverse transcriptase inhibitors (RTI) and protease inhibitors (PI) were analyzed according to mutations included in the surveillance drug-resistance mutations list updated in 2009. TC were defined as those comprising viruses from five or more individuals whose sequences clustered in maximum likelihood phylogenetic trees with a bootstrap value ≥90%. The overall prevalence of TDR to any drug was 9.9%: 4.9% to nucleoside RTIs (NRTIs), 3.6% to non-nucleoside RTIs (NNRTIs), and 2.7% to PIs. A significant decrease of TDR to NRTIs over time was observed [from 10% in 2004 to 2% in 2012 (p=0.01)]. Sixty eight (42.2%) of 161 sequences with TDR were included in 25 TC composed of 5 or more individuals. Of them, 9 clusters harbored TDR associated with high level resistance to antiretroviral drugs. T215D revertant mutation was transmitted in a large cluster comprising 25 individuals. The impact of epidemiological networks on TDR frequency may explain its persistence in newly diagnosed individuals. The knowledge of the populations involved in TC would facilitate the design of prevention programs and public health interventions.

Phylogeny and Phylogeography of a Recent HIV-1 Subtype F Outbreak among Men Who Have Sex with Men in Spain Deriving from a Cluster with a Wide Geographic Circulation in Western Europe.

We recently reported the rapid expansion of an HIV-1 subtype F cluster among men who have sex with men (MSM) in the region of Galicia, Northwest Spain. Here we update this outbreak, analyze near full-length genomes, determine phylogenetic relationships, and estimate its origin. For this study, we used sequences of HIV-1 protease-reverse transcriptase and env V3 region, and for 17 samples, near full-length genome sequences were obtained. Phylogenetic analyses were performed via maximum likelihood. Locations and times of most recent common ancestors were estimated using Bayesian inference. Among samples analyzed by us, 100 HIV-1 F1 subsubtype infections of monophyletic origin were diagnosed in Spain, including 88 in Galicia and 12 in four other regions. Most viruses (n = 90) grouped in a subcluster (Galician subcluster), while 7 from Valladolid (Central Spain) grouped in another subcluster. At least 94 individuals were sexually-infected males and at least 71 were MSM. Seventeen near full-length genomes were uniformly of F1 subsubtype. Through similarity searches and phylogenetic analyses, we identified 18 viruses from four other Western European countries [Switzerland (n = 8), Belgium (n = 5), France (n = 3), and United Kingdom (n = 2)] and one from Brazil, from samples collected in 2005-2011, which branched within the subtype F cluster, outside of both Spanish subclusters, most of them corresponding to recently infected individuals. The most probable geographic origin and age of the Galician subcluster was Ferrol, Northwest Galicia, around 2007, while the Western European cluster probably emerged in Switzerland around 2002. In conclusion, a recently expanded HIV-1 subtype F cluster, the largest non-subtype B cluster reported in Western Europe, continues to spread among MSM in Spain; this cluster is part of a larger cluster with a wide geographic circulation in diverse Western European countries.

Short communication: Molecular epidemiology, phylogeny, and phylodynamics of CRF63_02A1, a recently originated HIV-1 circulating recombinant form spreading in Siberia.

The HIV-1 epidemic in Russia is dominated by the former Soviet Union subtype A (A(FSU)) variant, but other genetic forms are circulating in the country. One is the recently described CRF63_02A1, derived from recombination between a CRF02_AG variant circulating in Central Asia and A(FSU), which has spread in the Novosibirsk region, Siberia. Here we phylogenetically analyze pol and env segments from 24 HIV-1 samples from the Novosibirsk region collected in 2013, with characterization of three new near full-length genome CRF63_02A1 sequences, and estimate the time of the most recent common ancestor (tMRCA) and the demographic growth of CRF63_02A1 using a Bayesian method. The analyses revealed that CRF63_02A1 is highly predominant in the Novosibirsk region (81.2% in pol sequences) and is transmitted both among injecting drug users and by heterosexual contact. Similarity searches with database sequences combined with phylogenetic analyses show that CRF63_02A1 is circulating in East Kazakhstan and the Eastern area of Russia bordering China. The analyses of near full-length genome sequences show that its mosaic structure is more complex than reported, with 18 breakpoints. The tMRCA of CRF63_02A1 was estimated around 2006, with exponential growth in 2008-2009 and subsequent stabilization. These results provide new insights into the molecular epidemiology, phylogeny, and phylodynamics of CRF63_02A1.

Identification of new and unusual rev and nef transcripts expressed by an HIV type 1 primary isolate.

We analyzed RNA splice site usage in three HIV-1 subtype B primary isolates through reverse transcriptase polymerase chain reaction (RT-PCR) amplification of spliced RNAs using a fluorescently labeled primer, with computerized size determination and quantification of PCR products, which were also identified by clone sequencing. In one isolate, P2149-3, unusual and unreported spliced transcripts were detected. This isolate preferentially used for rev RNA generation a 3' splice site (3'ss) located five nucleotides upstream of A4a, previously identified only in a T cell line-adapted virus and in a group O isolate, and designated A4d. P2149-3 also used an unreported 3'ss for rev RNA generation, designated A4h, located 20 nucleotides upstream of 3'ss A4c. Additionally, unusual nef RNAs using 3'ss A5a and A7a and with exon composition 1.3.7 were identified. The identification of several unusual and unreported spliced transcripts in an HIV-1 primary isolate suggests a greater diversity of splice site usage in HIV-1 than previously appreciated.

Simplified sequence-specific oligonucleotide-based polymerase chain reaction protocol to characterize human major histocompatibility complex A*02 and A*24 specificities.

Major histocompatibility complex (MHC) multimer technology is used in studies of high scientific and clinical interest for the identification, analysis, purification, and adoptive transfer of virus-specific T cells. MHC peptide multimers are usually specific for MHC A*02 or A*24 specificities because both specificities exhibit a high worldwide frequency. However, commercially available typing methods perform complete typing instead of browsing for these prevailing specificities. In this study we demonstrate an easy and accessible polymerase chain reaction-based method to accurately identify A*02 and A*24 samples in a cost-effective way.

Construction and phenotypic characterization of HIV type 1 functional envelope clones of subtypes G and F.

Subtype G has been estimated to represent the fourth most prevalent clade in the HIV-1 pandemic and subtype F is widely circulating in parts of South America (frequently within BF recombinant forms) and in Romania. However, functional envelope clones of these subtypes are lacking, which are needed for studies on antibody-mediated neutralization, coreceptor usage, and efficiency of viral entry inhibitor drugs. Here we report the construction, neutralization properties, and coreceptor usage of HIV-1 functional envelope clones of subtypes G (n = 15) and F (n = 7). These clones were obtained through RT-PCR amplification of HIV-1 gp160 from plasma RNA, and were used for pseudovirus production. All 15 subtype G-enveloped pseudoviruses were resistant to neutralization by gp120-targeted broadly neutralizing monoclonal antibodies (MAbs) b12 and 2G12, while a majority were neutralized by gp41-targeted MAbs 2F5 and 4E10. With regard to the subtype F envelopes, all seven pseudoviruses were resistant to 2F5 and b12, six were resistant to G12, and six were neutralized by 4E10. Coreceptor usage testing revealed that 21 of 22 envelopes were CCR5-tropic, including all 15 subtype G envelopes, seven of which were from patients with CD4(+) T cell counts <200/ml. These results confirm the broadly neutralizing activity of 4E10 on envelope clones across all tested group M clades, including subtypes G and F, reveal the resistance of most subtype F-enveloped pseudoviruses to broadly neutralizing MAbs b12, 2G12, and 2F5, and suggest that, similarly to subtype C, CXCR4 tropism is uncommon in subtype G, even at advanced stages of infection.

Relación de la asignación de personal de enfermería con indicadores de resultado de la calidad de la atención en unidades de cuidados intensivos de adultos / Association between Nurse Staffing and Indicators of Healthcare Quality in Adult Intensive Care Units / Relação da designação de pessoal de enfermagem com indicadores de resultado da qualidade do atendimento em unidades de tratamento intensivo de adultos

Objetivo: determinar la relación de la asignación de personal de enfermería con indicadores de resultado de la calidad de la atención en unidades de cuidados intensivos. Materiales y métodos: se hizo un estudio descriptivo con un componente correlacional, en el que se observaron 4006 turnos y se analizaron las variables de asignación de personal (estructura) y de resultado, en términos de casos de shocko paro, infecciones asociadas a la atención en salud, número de muertes y totalidad de eventos negativos en salud. Resultados: los resultados muestran que las enfermeras desarrollan un número reducido de actividades de cuidado directo al paciente, que se concentran en la administración de medicamentos y la supervisión de transfusiones, y otras tareas administrativas, mientras que el personal auxiliar se hace cargo del cuidado directo al paciente, incluidas las actividades complejas. La razón paciente/profesional de enfermería promedio fue de 5,4 pacientes por cada enfermera disponible en la unidad, y la razón paciente/auxiliar de enfermería promedio fue de 2,4. Las correlaciones no son concluyentes. Conclusiones: se sugiere estudiar con mayor profundidad el skill mix como una medida más sensible de asignación de personal, a la vez que se plantean recomendaciones en torno a la diferenciación de actividades entre profesionales y auxiliares de enfermería, y algunas consideraciones relativas a la asignación de personal.

Purpose: The purpose of the study is to identify the association between nursing staffing and indicators of healthcare quality in intensive care units. Materials and Methods: A descriptive study was done using a correlational component in which 4006 work shifts were observed and staffing variables (structure) and results were analyzed in terms of cases of shock or arrest, infections associated with health care, number of deaths, and all adverse health events. Findings: The findings show nurses conduct a limited number of activities involving direct patient care. These focus on administering medication, supervising transfusions and other administrative tasks, while assistant nurses handle direct patient care, including complex activities. The average patient-to-nurse ratio was 5.4 patients per nurse available during the shift, while the average patient-to-nursing assistant ratio was 2.4. The correlations are inconclusive. Conclusions: It is suggested the skill mix be explored further as a more sensitive measure of staffing, and recommendations are made on differentiating activities between nursing professionals and assistant nurses. Several considerations are raise concerning staff allocation.

Objetivo: determinar a relação da designação de pessoal de enfermagem com indicadores de resultado da qualidade do atendimento em unidades de tratamento intensivo. Materiais e métodos: realizou-se um estudo descritivo com um componente correlacional, no qual foram observados 4.006 turnos e analisadas as variáveis de designação de pessoal (estrutura) e de resultado, em termos de caso de choque ou parada, infecções associadas à atenção em saúde, número de mortes e totalidade de eventos negativos em saúde. Resultados: os resultados mostram que os enfermeiros desenvolvem um número reduzido de atividades de cuidado direto do paciente, que se concentram na administração de medicamentos e na supervisão de transfusões, além de outras tarefas administrativas, enquanto o pessoal auxiliar se responsabiliza pelo cuidado direto do paciente, o que inclui as atividades complexas. A razão paciente-profissional de enfermagem média foi de 5,4 pacientes por cada enfermeiro disponível na unidade, e a razão paciente-auxiliar de enfermagem foi de 2,4. As correlações não são conclusivas. Conclusões: sugere-se estudar com maior profundidade o skill mix como uma medida mais sensível de designação de pessoal, ao mesmo tempo que se propõem recomendações sobre a diferenciação de atividades entre profissionais e auxiliares de enfermagem, bem como algumas considerações relativas à designação de pessoal.

Calidad percibida de la atención de enfermería por pacientes hospitalizados / Perceived quality of nursing care for hospitalized patients

El objetivo de la presente investigación fue determinar la calidad percibida de la atención de Enfermería por pacientes hospitalizados en una Institución de Servicios de Salud de Barranquilla (Colombia). Material y Método: se llevó a cabo un estudio descriptivo, de corte transversal, cuantitativo, en el que se encuestaron 158 pacientes a través de un cuestionario de calidad percibida, previo cumplimiento de criterios de inclusión. Los datos se tabularon y graficaron en Microsoft Excel, utilizando medidas de tendencia central, y frecuencias absolutas y relativas; guardando el rigor ético y metodológico correspondiente. Resultados: El 44% fueron hombres y el 56% mujeres, con una media de 48.5 años. La calidad percibida de la atención de Enfermería es "Peor y mucho peor de lo que esperaba" en un 4%, "Como me lo esperaba" en un 17%, y "Mejor y mucho mejor de lo que me esperaba" en un 78%. El 48.7% estuvo "Muy Satisfecho", el 48.1% "Satisfecho", y el 3% "Poco y Nada Satisfecho". Conclusión: El 78% de los pacientes evidenció altos niveles de calidad percibida de la atención de Enfermería; el 17% modestos niveles de calidad percibida; y el 4% bajos niveles de calidad percibida. El 96.8% se sintió satisfecho y el 97% recomendaría el hospital.

The aim of this investigation was to determine the perceived quality of nursing care for patients hospitalized in an institution Health Services Barranquilla (Colombia). Method: A descriptive, cross-sectional, quantitative study, in which 158 patients were surveyed through a questionnaire of perceived quality, after fulfilling the inclusion criteria, was conducted. The data were tabulated and graphicated in Microsoft Excel, using central tendency measures and absolute and relative frequencies, keeping the appropriate ethical and methodological rigor. Results: 44% were male and 56% female, with a mean of 48.5 years old. The perceived quality of nursing care is "worse and much worse than I expected" by 4%, "As I expected" by 17%, and "Better, much better than I expected" by 78%. The 48.7% were "Very Satisfied", 48.1% "Satisfied", and 3% "Soon and Not Satisfied". Conclusion: 78% of patients showed high levels of perceived quality of nursing care, 17% modest levels of perceived quality, and 4% lower levels of perceived quality. The 96.8% was satisfied and 97% would recommend the hospital.

Molecular epidemiology of HIV-1 in St Petersburg, Russia: predominance of subtype A, former Soviet Union variant, and identification of intrasubtype subclusters.

OBJECTIVES: To examine HIV-1 genetic diversity in St. Petersburg. METHODS: Partial HIV-1 pol sequences from 102 plasma samples collected in 2006 were analyzed with a Bayesian phylogeny inference method. RESULTS: Subtype A, former Soviet Union (FSU) variant (AFSU), was the predominant clade (89.3%); other clades were subtypes B (9.7%) and F1 (1%). AFSU was predominant both among injecting drug users (98.2%) and heterosexually infected individuals (91.4%), whereas subtype B was more prevalent among homosexual men (75%). Within the AFSU variant, most sequences (93.5%) branched within 1 of 4 strongly supported subclusters. The largest comprised 63% AFSU viruses and was uncommon outside St Petersburg. A second subcluster (17.4% AFSU viruses) corresponds to the variant with the V77I substitution in protease, which is widely circulating in different FSU countries. Two minor subclusters comprised 8.7% and 6.5% AFSU viruses, respectively. There was no correlation between risk exposure and AFSU subclusters. Six of 8 subtype B sequences, 4 of them from homosexual men, grouped in a monophyletic subcluster. CONCLUSIONS: The results of this study show a great predominance of AFSU viruses in St Petersburg and point to a few phylogenetically identifiable introductions as the origin of most current HIV-1 AFSU infections in the city.

La administración que se enseña en el pregrado de enfermería / Administration learning in nursing undergraduate programs / A administração que se ensina nos curso de pré-graduação de enfermagem

La enseñanza de la administración, gestión o gerencia1 se ha incorporado en la formación del pregrado de enfermería en distintos momentos, y ha estado ligada al desarrollo de los países y de la profesión, a la apropiación y evolución del conocimiento administrativo y a las necesidades del contexto. Este estudio descriptivo se propuso identificar las temáticas de administración- gestión o gerencia que se imparten actualmente en programas de enfermería en Latinoamérica, y los objetivos educacionales o competencias de formación que se han propuesto para establecer, a partir de los hallazgos, algunos lineamientos para la enseñanza de las ciencias administrativas aplicadas a enfermería. Las encuestas que respondieron 15 personas en representación del programa que dicta la unidad docente de la región: 10 colombianas y 5 extranjeras, mostraron que las ciencias administrativas se enseñan a través de 31 asignaturas; en la mayoría de ellas se contempla la formación en temáticas relacionadas con enfoques administrativos -21-, seguidas de proceso administrativo y aplicación a enfermería -20-, contexto para la aplicación en salud y enfermería -17-, pensamiento administrativo -9- y, en una menor proporción, temas como legislación en salud, ética y empresa. Un alto porcentaje (77 %) contemplan objetivos de información y formación; el 23 % describe competencias sobre lo técnico y lo metodológico. El análisis de los hallazgos, contrastados con la situación del sector salud, guía la propuesta de enseñanza a fortalecer la aplicación de las herramientas administrativas en la gestión profesional, teniendo en cuenta la normatividad, pero en ningún caso se condiciona el desarrollo de la profesión a los requerimientos del contexto.

Administration and management subjects have been gradually included in the nursing undergraduate programs at different times. This is closely linked to the development of the countries and the profession, the appropriation and evolution of administrative knowledge and the needs of the context. This descriptive study is aimed to identify the administration and management themes that are currently taught in nursing programs in Latin America, and the educational objectives or capacity-building competences proposed to set, based on our findings, some guidelines for teaching administrative sciences applied to nursing. The surveys responded in 15 teaching units of the region: 10 in Colombia and 5 abroad, showed that administrative sciences are taught through 31 subjects; that encompass, primarily, education in themes related to administrative approaches -21-, followed by administrative processes and application to the nursing -20-, context for applying administration in health and nursing -17-, administrative thinking -9- and, to a lesser extent, subjects dealing with health care legislations, ethics and entrepreneurship. Seventy-seven per cent (77 %) includes information and formation objectives; 23% describes competences about technical and methodological aspects. The analysis of data, compared against the situation of the health sector, guides the teaching proposal to strengthen the use of administrative tools in nursing management, considering regulatory issues, but under no circumstance, the development of the professions is subject to the requirement of the context.

Oensino da administração, gestão ou gerência tem sido incorporado na formação dos cursos de pré-graduação de enfermagem em diferentes momentos, e tem estado vinculado ao desenvolvimento dos países e da profssão, à apropriação e evolução do conhecimento administrativo e às necessidades do contexto. Este estudo descritivo objetivou identificar as temáticas de administração - gestão ou gerência que são ensinadas atualmente em programas de enfermagem na América Latina, e os objetivos educativos ou competências de formação propostas para estabelecer, com base nos achados, algumas linhas básicas para o ensino das ciências administrativas aplicadas à enfermagem. As enquetes que responderam em 15 unidades docentes da região: 10 colombianas e 5 estrangeiras, demonstraram que as ciências administrativas são ensinadas através de 31 matérias; que contemplam, maiormente, a formação em temáticas relativas às abordagens administrativas -21-, seguidas de processos administrativos e aplicação à enfermagem -20-, contexto para a aplicação em saúde e enfermagem -17-, pensa-mento administrativo -9- e, em menor proporção, temas com legislação em saúde, ética e empresa. Uma alta porcentagem (77 %) contempla objetivos de informação e formação; 23 % descreve competências sobre aspetos técnicos e metodológicos. A análise dos dados, confrontados com a situação do setor saúde, orienta a proposta de ensino para fortalecer a aplicação de ferramentas administrativas na gestão professional, levando em conta a regulamentação, mas sob nenhuma circunstância, o desenvolvimento da profissão é sujeito aos requerimentos do contexto.

Oligonucleotide ligation assay for detection of mutations associated with reverse transcriptase and protease inhibitor resistance in non-B subtypes and recombinant forms of human immunodeficiency virus type 1.

The oligonucleotide ligation assay is a genotypic assay for the detection of resistance-associated mutations to reverse transcriptase and protease inhibitors in human immunodeficiency virus type 1 subtype B. This assay has been modified and developed for non-B subtypes and recombinant strains and has been evaluated with sequencing, resulting in a more sensitive assay than sequencing for non-B subtypes.

Negative control of Listeria monocytogenes virulence genes by a diffusible autorepressor.

Virulence genes from the facultative intracellular pathogen Listeria monocytogenes are controlled by the transcriptional regulator PrfA. Although PrfA synthesis is activated at 37 degrees C, PrfA-dependent expression remains low in rich medium. However, a strong induction of the PrfA regulon is observed when L. monocytogenes is cultured in the presence of activated charcoal. Here, we show that the 'charcoal effect' results from the adsorption of a diffusible autorepressor substance released by L. monocytogenes during exponential growth. Analyses using an L. monocytogenes strain in which the prfA gene is expressed constitutively at 37 degrees C from a plasmid indicate that the autoregulatory substance represses PrfA-dependent expression by inhibiting PrfA activity. PrfA presumably functions via an allosteric activation mechanism. The inhibitory effect is bypassed by a PrfA* mutation that locks PrfA in fully active conformation, suggesting that the autorepressor interferes with the allosteric shift of PrfA. Our data indicate that the listerial autorepressor is a low-molecular-weight hydrophobic substance. We suggest that this diffusible substance mediates a quorum-sensing mechanism by which L. monocytogenes restricts the expression of its PrfA virulence regulon. This autoregulatory pathway could serve L. monocytogenes to ensure the silencing of virulence genes during extracellular growth at 37 degrees C. It may also play a role during intracellular infection, by limiting the damage to the host cell caused by an excess production of cytotoxic PrfA-dependent virulence factors in the PrfA-activating cytosolic compartment.