RESUMEN
Resumen. El Insomnio infantil definido como la dificultad mantenida, a pesar de la oportunidad y en función etaria, para iniciar o mantener el sueño o su calidad que provoca alteraciones funcionales en el niño y/o familia. Puede repercutir significativamente en la conducta, aprendizaje y metabolismo del niño y en su familia afectando su calidad de vida. La actigrafía nos permite a través de un dispositivo identificar periodos de vigilia y sueño. El objetivo de este trabajo es caracterizar a través de la actigrafía el patrón sueño-vigilia y evaluar la exposición a luz azul en niños menores de 2 años que consultan por insomnio. Se realizó un estudio observacional, descriptivo, transversal de lactantes derivados por insomnio al Centro de Sueño de Red de Salud UC Christus, durante: Marzo 2017-2018, con actigrafía. Fueron 20 actigrafías de 7 días. Edades entre 5 y 22 meses con diagnóstico de insomnio que no respondió al manejo inicial. 8 hombres, 12 mujeres. Horarios promedios: Acostarse: 20:31 hrs. Levantarse: 7:43 hrs. Horarios variables para acostarse: 15/20. 10/20 siestas después de las 16 hrs. Todos presentaron tiempo total de sueño disminuido, con aumentos del tiempo despierto una vez iniciado el sueño. Latencias del sueño aumentadas: 6/20. Eficiencia del sueño disminuidas en 4/20. Despertares nocturnos: promedio: 10.58. Expuestos a luz azul: 14/20. Horas exposición media: 2,4 hr/evento. Concluimos de este estudio que las principales dificultades fueron los despertares nocturnos con largos tiempo de vigilia, con disminución del tiempo total de sueño para la edad, y la actigrafía fue una herramienta de apoyo para objetivar conductas que dificultan la adquisición de un buen patrón de sueño.Palabras Clave: Insomnio infantil, sueño en lactantes, actigrafía, luz azul, despertares.
Abstract. Child insomnia is defined as sustained difficulty, despite the opportunity and according to age group, to initiate or maintain sleep, or a sleep quality causing that causes alterations in the child or family. It can significantly impact behavior, learning and metabolism of the child and his or her family, affecting their quality of life. The actigraphy through a device allows us to identify periods of wakefulness and sleep. The purpose of this work is to characterize sleep-wake patterns through actigraphy and to determine exposure to blue light in children younger than 2 years old, consulting for insomnia. An observational, descriptive, cross-sectional study of infants consulting for insomnia at the Sleep Center of UC Christus health network, in the period from March 2017 to 2018, with actigraphy. Twenty week long actigraphies were performed. The ages of the patients varied between 5 and 22 months, all had a diagnosis of insomnia that did not respond to initial management. Eight patients were male and 12 female. Hourly averages: bedtime: 8:31 PM. Waking up: 7:43 AM. Time variable for bedtime: 15/20. 10/20 NAPs after 16 hrs. All showed decreased sleep, with increases of total awake time once sleep began. Increased sleep latency: 6/20. Sleep efficiency decreased in 4/20. Nighttime Awakenings: average: 10.58. Exposed to blue light: 14/20. Average exposure in hours: 2.4 hr/event. We conclude from this study that the main difficulties were the nighttime awakenings with long time vigil, with decrease of the total sleep time for the age, and the actigraphy was a support tool to record behaviors that hinder a normal sleep pattern acquisition.Key words: Child insomnia, sleep in infants, actigraphy, blue light, awakenings
Asunto(s)
Humanos , Masculino , Femenino , Lactante , Actigrafía/métodos , Trastornos del Inicio y del Mantenimiento del Sueño/diagnóstico , Luz AzulRESUMEN
BACKGROUND: Variants in HIV-coreceptor C-C chemokine receptor type 5 (CCR5) and Human leukocyte antigen (HLA) genes are the most important host genetic factors associated with HIV infection and disease progression. Our aim was to analyze the association of these genetic factors in the presence of clinical symptoms during Primary HIV Infection (PHI) and disease progression within the first year. METHODS: Seventy subjects diagnosed during PHI were studied (55 symptomatic and 15 asymptomatic). Viral load (VL) and CD4 T-cell count were evaluated. HIV progression was defined by presence of B or C events and/or CD4 T-cell counts <350 cell/mm3. CCR5 haplotypes were characterized by polymerase chain reaction and SDM-PCR-RFLP. HLA-I characterization was performed by Sequencing. RESULTS: Symptoms during PHI were significantly associated with lower frequency of CCR5-CF1 (1.8% vs. 26.7%, p = 0.006). Rapid progression was significantly associated with higher frequency of CCR5-CF2 (16.7% vs. 0%, p = 0.024) and HLA-A*11 (16.7% vs. 1.2%, p = 0.003) and lower frequency of HLA-C*3 (2.8% vs. 17.5%, p = 0.035). Higher baseline VL was significantly associated with presence of HLA-A*11, HLA-A*24, and absence of HLA-A*31 and HLA-B*57. Higher 6-month VL was significantly associated with presence of CCR5-HHE, HLA-A*24, HLA-B*53, and absence of HLA-A*31 and CCR5-CF1. Lower baseline CD4 T-cell count was significantly associated with presence of HLA-A*24/*33, HLA-B*53, CCR5-CF2 and absence of HLA-A*01/*23 and CCR5-HHA. Lower 6-month CD4 T-cell count was associated with presence of HLA-A*24 and HLA-B*53, and absence of HLA-A*01 and HLA-B*07/*39. Moreover, lower 12-month CD4 T-cell count was significantly associated with presence of HLA-A*33, HLA-B*14, HLA-C*08, CCR5-CF2, and absence of HLA-B*07 and HLA-C*07. CONCLUSION: Several host factors were significantly associated with disease progression in PHI subjects. Most results agree with previous studies performed in other groups. However, some genetic factor associations are being described for the first time, highlighting the importance of genetic studies at a local level.
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Seropositividad para VIH/genética , Antígenos HLA/genética , Factores Celulares Derivados del Huésped/metabolismo , Receptores CCR5/genética , Argentina , Western Blotting , Recuento de Linfocito CD4 , Progresión de la Enfermedad , Haplotipos/genética , Humanos , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Carga ViralRESUMEN
The HIV-1 epidemic in South America is dominated by pure subtypes (mostly B and C) and more than 7 BF and BC recombinant forms. In Argentina, circulating recombinant forms (CRFs) comprised of subtypes B and F make up more than 50% of HIV infections. For this study, 28 HIV-1 primary isolates were obtained from patients in Buenos Aires, Argentina and initially classified into subtype B (nâ=â9, 32.1%), C (nâ=â1, 3.6%), and CRFs (nâ=â18, 64.3%) using partial pol and vpu-env sequences, which proved to be inconsistent and inaccurate for these phylogenetic analyses. Near full length genome sequences of these primary HIV-1 isolates revealed that nearly all intersubtype BF recombination sites were unique and countered previous "CRF" B/F classifications. The majority of these Argentinean HIV-1 isolates were CCR5-using but 4 had a dual/mixed tropism as predicted by both phenotypic and genotypic assays. Comparison of the replicative fitness of these BF primary HIV-1 isolates to circulating B, F, and C HIV-1 using pairwise competitions in peripheral blood mononuclear cells (PBMCs) indicated a similarity in fitness of these BF recombinants to subtypes B and F HIV-1 (of the same co-receptor usage) whereas subtype C HIV-1 was significantly less fit than all as previously reported. These results suggest that the multitude of BF HIV-1 strains present within the Argentinean population do not appear to have gained replicative fitness following recent B and F recombination events.
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VIH-1/genética , VIH-1/patogenicidad , Recombinación Genética/genética , Argentina/epidemiología , Genoma Viral/genética , Infecciones por VIH/epidemiología , VIH-1/clasificación , Humanos , Filogenia , Replicación Viral/genética , Replicación Viral/fisiologíaRESUMEN
The internal transcribed spacer (ITS) regions of rDNA genes of 49 Histoplasma capsulatum (48 from clinical samples and one from soil) isolates were examined. Nucleotide sequence heterogeneity within this region was useful for phylogenetic classification of H. capsulatum and species identification. Thus, in 45 of 49 isolates we observed higher percentages of identity in the nucleotide sequences of ITS regions when the isolates studied herein were compared with those reported in our country in the South America B clade. Phylogenetic analyses of rDNA sequences corresponding to the 537 bp of the ITS region obtained from H. capsulatum isolates assigned South America type B clade (45 isolates), North America type 1 and Asia clade (2 isolates each one). H. capsulatum strains isolated from soil and from patients living in Argentina (45 of 49) clustered together with the H. capsulatum isolates of the South America B clade. The high level of genetic similarity among our isolates suggests that almost one genetic population is present in the microenvironment. Isolates described as H. capsulatum var. capsulatum or var. farciminosum (2 isolates) did not form a monophyletic group and were found in the Asia clade. Subsequent studies are needed to properly identify these isolates.
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ADN Espaciador Ribosómico/genética , Variación Genética , Histoplasma/genética , Histoplasma/aislamiento & purificación , Histoplasmosis/microbiología , Histoplasmosis/veterinaria , Animales , Argentina/epidemiología , Secuencia de Bases , ADN de Hongos/genética , Histoplasma/clasificación , Histoplasmosis/epidemiología , Enfermedades de los Caballos/microbiología , Caballos , Humanos , Datos de Secuencia Molecular , FilogeniaRESUMEN
El propósito de esta presentación es dar a conocer los hallazgos clínicos, micológicos e histopatológicos de una aspergilosis pulmonar crónica por Aspergillus nomius, en una mujer de 52 años de edad que sufre de una feohifomicosis diseminada por Exophiala spinifera, con lesiones cutáneas, ganglionares y óseas de 22 años de evolución. La aspergilosis pulmonar se presentó como una neumopatía crónica, de 3 años de duración, que evolucionó hacia la abscedación. Esta infección fúngica se produjo durante el tratamiento con 800 mg/diarios de posaconazol y respondió favorablemente a la administraciónconjunta de caspofungina por vía intravenosa. Finalmente, la enferma fue intervenida quirúrgicamente y se le extirparon los lóbulos medio e inferior del pulmón derecho. En el estudio histopatológico de la pieza quirúrgica se comprobó que Aspergillus nomius invadía los vasos sanguíneos y que se formaba un granuloma epitelioide con células gigantes en torno a las hifas endovasculares. El agente causal deeste caso se aisló de múltiples muestras de expectoración y lavados broncoalveolares, así como de lapieza quirúrgica. Su ubicación taxonómica se hizo en base a estudios de biología molecular. No pudo establecerse una causa clara de inmunodeficiencia en este caso.
Asunto(s)
Humanos , Femenino , Persona de Mediana Edad , Antifúngicos/administración & dosificación , Aspergillus , Aspergilosis Broncopulmonar AlérgicaRESUMEN
Proteolysis of sperm histones in the sea urchin male pronucleus is the consequence of the activation at fertilization of a maternal cysteine protease. We previously showed that this protein is required for male chromatin remodelling and for cell-cycle progression in the newly formed embryos. This enzyme is present in the nucleus of unfertilized eggs and is rapidly recruited to the male pronucleus after insemination. Interestingly, this cysteine-protease remains co-localized with chromatin during S phase of the first cell cycle, migrates to the mitotic spindle in M-phase and is re-located to the nuclei of daughter cells after cytokinesis. Here we identified the protease encoding cDNA and found a high sequence identity to cathepsin proteases of various organisms. A phylogenetical analysis clearly demonstrates that this sperm histone protease (SpHp) belongs to the cathepsin L sub-type. After an initial phase of ubiquitous expression throughout cleavage stages, SpHp gene transcripts become restricted to endomesodermic territories during the blastula stage. The transcripts are localized in the invaginating endoderm during gastrulation and a gut specific pattern continues through the prism and early pluteus stages. In addition, a concomitant expression of SpHp transcripts is detected in cells of the skeletogenic lineage and in accordance a pharmacological disruption of SpHp activity prevents growth of skeletal rods. These results further document the role of this nuclear cathepsin L during development.
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Catepsina L/metabolismo , Endopeptidasas/metabolismo , Histonas/metabolismo , Erizos de Mar/embriología , Erizos de Mar/enzimología , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Catepsina L/análisis , Catepsina L/genética , ADN Complementario/genética , Endopeptidasas/análisis , Endopeptidasas/genética , Fertilización , Regulación del Desarrollo de la Expresión Génica , Masculino , Datos de Secuencia Molecular , Filogenia , Erizos de Mar/citología , Erizos de Mar/genética , Alineación de Secuencia , Espermatozoides/metabolismoRESUMEN
HIV-1 epidemics in South America are believed to have originated in part from the subtype B epidemic initiated in the Caribbean/North America region. However, circulation of BF recombinants in similar proportions was extensively reported. Information currently shows that many BF recombinants share a recombination structure similar to that found in the CRF12_BF. In the present study, analyzing a set of 405 HIV sequences, we identified the most likely origin of the BF epidemic in an early event of recombination. We found that the subtype B epidemics in South America analyzed in the present study were initiated by a founder event that occurred in the early 1970s, a few years after the introduction of these strains in the Americas. Regarding the F/BF recombinant epidemics, by analyzing a subtype F genomic segment within the viral gene gag present in the majority of the BF recombinants, we found evidence of a geographic divergence very soon after the introduction of subtype F strains in South America. Moreover, through analysis of a subtype B segment present in all the CRF12_BF-like recombination structure, we estimated the circulation of the subtype B strain that gave rise to that recombinant structure around the same time period estimated for the introduction of subtype F strains. The HIV epidemics in South America were initiated in part through a founder event driven by subtype B strains coming from the previously established epidemic in the north of the continent. A second introduction driven by subtype F strains is likely to have encountered the incipient subtype B epidemic that soon after their arrival recombined with them, originating the BF epidemic in the region. These results may explain why in South America the majority of F sequences are found as BF recombinants.
Asunto(s)
Evolución Molecular , Infecciones por VIH/epidemiología , Infecciones por VIH/virología , VIH-1/clasificación , VIH-1/genética , Recombinación Genética , Análisis por Conglomerados , Genoma Viral , Genotipo , VIH-1/aislamiento & purificación , Humanos , Epidemiología Molecular , Filogenia , Análisis de Secuencia de ADN , América del Sur/epidemiología , Factores de TiempoRESUMEN
BACKGROUND: Cytotoxic T-Lymphocyte (CTL) response drives the evolution of HIV-1 at a host-level by selecting HLA-restricted escape mutations. Dissecting the dynamics of these escape mutations at a population-level would help to understand how HLA-mediated selection drives the evolution of HIV-1. METHODOLOGY/PRINCIPAL FINDINGS: We undertook a study of the dynamics of HIV-1 CTL-escape mutations by analyzing through statistical approaches and phylogenetic methods the viral gene gag sequenced in plasma samples collected between the years 1987 and 2006 from 302 drug-naïve HIV-positive patients. By applying logistic regression models and after performing correction for multiple test, we identified 22 potential CTL-escape mutations (p-value<0.05; q-value<0.2); 10 of these associations were confirmed in samples biologically independent by a Bayesian Markov Chain Monte-Carlo method. Analyzing their prevalence back in time we found that escape mutations that are the consensus residue in samples collected after 2003 have actually significantly increased in time in one of either B or F subtype until becoming the most frequent residue, while dominating the other viral subtype. Their estimated prevalence in the viral subtype they did not dominate was lower than 30% for the majority of samples collected at the end of the 80's. In addition, when screening the entire viral region, we found that the 75% of positions significantly changing in time (p<0.05) were located within known CTL epitopes. CONCLUSIONS: Across HIV Gag protein, the rise of polymorphisms from independent origin during the last twenty years of epidemic in our setting was related to an association with an HLA allele. The fact that these mutations accumulated in one of either B or F subtypes have also dominated the other subtype shows how this selection might be causing a convergence of viral subtypes to variants which are more likely to evade the immune response of the population where they circulate.
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VIH-1/genética , Antígenos HLA/inmunología , Mutación , Selección Genética , Antígenos Virales/genética , Evolución Biológica , Epítopos/genética , Productos del Gen gag/genética , Productos del Gen gag/inmunología , VIH-1/inmunología , Antígenos HLA-B/inmunología , Humanos , Inmunidad , Modelos Estadísticos , Linfocitos T Citotóxicos/inmunologíaRESUMEN
BACKGROUND: HIV-1 is characterized by its rapid genetic evolution and high diversity as a consequence of its error-prone reverse transcriptase and genetic recombination. This latter mechanism is responsible for the creation of circulating recombinant forms (CRFs) found in nature. Previous studies from our lab group have shown that the epidemic in Argentina is characterized by one highly prevalent circulating recombinant form, CRF12_BF, and many related BF recombinant forms. Since transcriptional transactivation of the HIV-1 long terminal repeat (LTR) promoter element requires the essential viral Tat protein, since these genetic structures underwent recombination in variants widely spread in South America, the aim of this work was to study transcriptional activity associated with the recombinant LTR and Tat elements. RESULTS: Differential transcriptional activity was measured for the BF recombinant LTR/Tat complex that is present in widely spread viral variants was demonstrated. This analysis demonstrated a higher activity for the BF complex when compared to its B subtype counterpart. CONCLUSION: This study indicates structural and functional consequences of recombination events within the LTR promoter and Tat transactivator protein of a naturally occurring HIV-1 recombinant form.
Asunto(s)
Variación Genética , Duplicado del Terminal Largo de VIH/genética , VIH-1/genética , Activación Transcripcional , Síndrome de Inmunodeficiencia Adquirida/virología , Adulto , Línea Celular , Niño , Clonación Molecular , ADN Viral/genética , Productos del Gen tat/genética , Genes Reporteros , VIH-1/clasificación , VIH-1/aislamiento & purificación , Humanos , Recombinación Genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transfección , Productos del Gen tat del Virus de la Inmunodeficiencia HumanaRESUMEN
UNLABELLED: A total of 220 individuals were included in this study, 112 HIV-seronegative healthy individuals and 108 HIV-1-infected patients involving: 18 AIDS patients with Toxoplasmic encephalitis (AIDS-TE), 49 AIDS patients without TE, and 41 asymptomatic patients, were genotyping for DR and DQ loci by molecular biology techniques. Fisher's Exact test was used for statistical analysis. HLA-DQB*0402 and DRB1*08 alleles were associated with a high risk to develop opportunistic infections with neurological involvement, mainly Toxoplasma encephalitis in relationship with subjects healthy (OR = 20.43; Pc = 7.0 x 10(-6) and OR = 11; Pc = 2.6 x 10(-4), respectively); in relationship with AIDS no TE (OR = 6.98; Pc = 0.028 and OR = 4.85; P = 0.012, Pc = 0.14) and with patients in asymptomatic stage (OR = 61.50, Pc = 8.4 x 10(-6) and OR = 19.38; Pc = 3.9 x 10(-4)), respectively. CONCLUSIONS: It was concluded that the presence of HLA-DQB*0402 and DRB1*08 alleles in HIV-1-positive patients could be considered risk factors for developing neurological opportunistic infections, mainly Toxoplasmic encephalitis.
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Infecciones Oportunistas Relacionadas con el SIDA/genética , Encefalitis/genética , Antígenos HLA-DQ/genética , Antígenos HLA-DR/genética , Toxoplasmosis Cerebral/genética , Infecciones Oportunistas Relacionadas con el SIDA/complicaciones , Argentina/epidemiología , Estudios de Casos y Controles , Encefalitis/complicaciones , Frecuencia de los Genes , Predisposición Genética a la Enfermedad , VIH-1 , Antígenos HLA-DQ/clasificación , Cadenas beta de HLA-DQ , Antígenos HLA-DR/clasificación , Humanos , Fenotipo , Toxoplasmosis Cerebral/complicacionesRESUMEN
Different complex structures of the pol gene have been identified in 284 HIV-1 B/F recombinant sequences obtained from a group of 587 patients under treatment failure from Argentina. To analyze the mosaic structures of these viral sequences and to determine their phylogenetic relationship, the 284 partial pol gene sequences of BF recombinant viruses were amplified by RT-PCR and sequenced. Intersubtype breakpoints were analyzed by bootscanning. Phylogenetic relationships were determined by means of neighbor-joining trees. The analysis of the sequences showed multiple phylogenetic topologies clustering within intersubtype BF reference sequences. At least three different mosaic patterns were found compared to previously described BF-type viruses with unequal distribution in the studied population. The analysis also showed that HIV-1 BF recombinant viruses with diverse mosaic structures are phylogenetically related in their F segments and in selected B fragments with the F1 subtype and with BF recombinant viruses from Brazil, respectively, suggesting a common recombinant ancestor. No association was observed between the prevalence of each mosaic pattern and the frequency of major drug-resistance mutations in PR and RT.
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Fármacos Anti-VIH/uso terapéutico , Genes pol/genética , Infecciones por VIH/tratamiento farmacológico , VIH-1/clasificación , Recombinación Genética , Inhibidores de la Transcriptasa Inversa/uso terapéutico , Fármacos Anti-VIH/farmacología , Argentina , Farmacorresistencia Viral , Quimioterapia Combinada , Infecciones por VIH/virología , Proteasa del VIH/genética , Transcriptasa Inversa del VIH/genética , VIH-1/efectos de los fármacos , VIH-1/genética , Humanos , Datos de Secuencia Molecular , Mutación , Filogenia , Inhibidores de la Transcriptasa Inversa/farmacología , Análisis de Secuencia de ADN , Insuficiencia del TratamientoRESUMEN
To investigate the immunopathogenic mechanisms of type I autoimmune hepatitis in children, we analyzed by quantitative or semiquantitative reverse transcription-polymerase chain reaction the expression of cytokines interferon (IFN)-gamma, interleukin (IL)-12p40, IL-18, IL-4, IL-10, and IL-12R beta 2. In addition, liver and peripheral blood was collected to investigate the expression of the natural killer T (NKT) cell marker V alpha 24. The presence of NKT cells in hepatic lesions were also identified by immunohistochemistry. The analysis was performed on liver biopsies from 25 children with type I autoimmune hepatitis. As disease controls, we included six children with hepatitis C virus-related chronic hepatitis and nine control livers. The expression of IFN-gamma and IL-12p40 was not detected in controls but was clearly upregulated in pathologic biopsies. In addition, these samples showed an increased expression of IL-18 (p = 0.0003), IL-4 (p = 0.0055), and IL-12R beta 2 (p = 0.007). Western blot analysis confirmed the expression of IL-12p40 and IL-18. However, for IL-18, we detected only the immature biologically inactive polypeptide. The V alpha 24 transcripts were found increased in the liver (p = 0.0007) where V alpha 24(+) cells were also localized, but decreased in peripheral blood mononuclear cells (p = 0.041). In addition to a type I immune response, NKT cells might play a substantial role in the pathogenesis of type I autoimmune hepatitis in children.
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Citocinas/genética , Expresión Génica , Hepatitis Autoinmune/patología , Interleucina-4/genética , Células TH1/inmunología , Adolescente , Autoanticuerpos/sangre , Biopsia con Aguja , Análisis Químico de la Sangre , Western Blotting , Niño , Citocinas/inmunología , Citocinas/metabolismo , Femenino , Hepatitis Autoinmune/genética , Hepatitis Autoinmune/inmunología , Humanos , Inmunohistoquímica , Interferón gamma/genética , Interleucina-10/genética , Interleucina-10/metabolismo , Interleucina-12/genética , Interleucina-12/metabolismo , Subunidad p40 de la Interleucina-12 , Interleucina-18/genética , Interleucina-18/metabolismo , Interleucina-4/inmunología , Interleucina-4/metabolismo , Células Asesinas Naturales/inmunología , Células Asesinas Naturales/metabolismo , Células Asesinas Naturales/patología , Leucocitos Mononucleares/química , Hígado/inmunología , Hígado/metabolismo , Hígado/patología , Masculino , Subunidades de Proteína/genética , Subunidades de Proteína/metabolismo , Receptores de Antígenos de Linfocitos T alfa-beta/análisis , Receptores de Antígenos de Linfocitos T alfa-beta/genética , Receptores de Interleucina/genética , Receptores de Interleucina/metabolismo , Receptores de Interleucina-12 , Linfocitos T Colaboradores-Inductores/inmunología , Linfocitos T Colaboradores-Inductores/metabolismo , Células TH1/metabolismoRESUMEN
To monitor HIV-1 diversity in Argentina, a phylogenetic-based analysis of HIV-1 partial pol sequences obtained for resistance testing in 587 treatment failure patients was performed in Buenos Aires city between 2001 and 2003. HIV-1 RNA was isolated from plasma samples and partial pol fragments amplified by RT-PCR. Sequences were obtained by automated sequencing. Phylogenetic analysis was performed and recombination patterns characterized. A total of 299 sequences grouped into clade B (50.94%) and 284 were B/F recombinants (48.38%). Four sequences were grouped into clades A, C, and F (0.68%). The clade C sample, 96105, was found to be a BC recombinant and samples 103396 and 104575 showed the same mosaic pattern with Kisii5009 from Kenya and 97KR004 from Korea, previously described as A2D recombinants. With the presence of two full-length genomes, one from Kenya and one from Korea, and now two partial genomes from Argentina, this recombinant is designated CRF16_A2D. Its presence on three continents shows that CRF16_A2D has a global distribution.
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Fármacos Anti-VIH/farmacología , Farmacorresistencia Viral/genética , Infecciones por VIH/virología , VIH-1/efectos de los fármacos , Análisis por Conglomerados , Genes pol , Infecciones por VIH/epidemiología , VIH-1/clasificación , VIH-1/genética , Humanos , Pruebas de Sensibilidad Microbiana , Filogenia , Recombinación GenéticaRESUMEN
Apoptosis of enterocytes is a feature that characterises the development of lesions in coeliac disease (CD). However, the intracellular pathways that lead to apoptosis of enterocytes have not been completely clarified. Bak is a member of the Bcl-2 family of proteins that acts as an endogenous promoter of apoptosis in normal enterocytes. However, its role in coeliac lesions has not been explored. We used small intestinal mucosa from patients with CD to evaluate the differential expression of members of the Bcl-2 family of proteins. Gene expression of Bak was analysed by RT-PCR of biopsies from 14 patients with untreated CD and from 19 controls without CD. In these samples, we also investigated the localisation of the Bak protein by immunohistochemistry and its apoptotic activity. In patients with untreated CD there was a 2.3-fold higher expression of Bak mRNA (p = 0.026), without significant differences in the expression of related genes bax or bcl-2. The higher expression of interferon gamma (IFN-gamma) (p = 0.036) and the higher number of apoptotic cells identified by the TUNEL method (p = 0.032) confirmed the proapoptotic status in the intestinal mucosa of CD patients. We found a significant positive correlation (p < 0.0001) between the expression of IFN-gamma and Bak mRNA in patients with untreated CD. The expression of Bak protein was higher in patients with CD, and the immunoreactivity was almost restricted to the epithelium. We found that Bak mRNA and its protein were overexpressed in the intestinal lesions of CD patients and that IFNgamma confers increased susceptibility for enterocytes to undergo apoptosis via upregulation of Bak.
Asunto(s)
Enfermedad Celíaca/genética , Enfermedad Celíaca/metabolismo , Proteínas de la Membrana/genética , Proteínas de la Membrana/metabolismo , Apoptosis , Estudios de Casos y Controles , Enfermedad Celíaca/patología , Niño , Preescolar , Duodeno/metabolismo , Duodeno/patología , Enterocitos/metabolismo , Enterocitos/patología , Expresión Génica , Humanos , Inmunohistoquímica , Lactante , Interferón gamma/genética , Mucosa Intestinal/metabolismo , Mucosa Intestinal/patología , Yeyuno/metabolismo , Yeyuno/patología , ARN Mensajero/genética , ARN Mensajero/metabolismo , Proteína Destructora del Antagonista Homólogo bcl-2RESUMEN
La instauración de tratamientos antirretrovirales de alta eficacia (HAARTs) ha llevado a la disminución de la morbimortalidad asociada a HIV-1/SIDA en países industrializados. Sin embargo, uno de los principales factores asociados a la falla terapéutica es la selección de variantes de HIV-1 resistentes a las drogas antirretrovirales (ARVs). Dichas variantes virales se caracterizan por presentar mutaciones en las enzimas virales que son los blancos de la acción de los fármacos: la proteasa viral (PR) y la transcriptasa reversa (RT). Las variantes del HIV-1 resitente a ARVs seleccionadas en pacientes tratados no sólo se asocian al fracaso terapéutico en dichos individuos, sino que también pueden ser transmitidas a la población no infectada. De esta forma, individuos infectados con HIV-1 que nunca han sido expuestos a drogas ARVs (naïve) pueden albergar virus resistentes, lo que llevaría a la falla terapéutica al iniciarse el tratamiento. El objetivo del presente trabajo fue el de caracterizar los perfiles de resistencia en pacientes HIV-1 seropositivos con falla terapéutica. Asimismo, con el fin de estimar la tasa de transmisión de variantes resistentes a ARVs, se estudiaron los perfiles de resistencia en individuos infectados con HIV-1 que no habían recibido tratamiento. Se estudiaron 399 pacientes HIV-1 seropositivos con falla terapéutica y 94 individuos naive infectados con HIV-1. Las muestras de plasma fueron recolectadas entre 1997 y 2001 de individuos de la Cdad. de Buenos Aires y sus alrededores. Se amplificaron las regiones de la PR y RT del HIV-1 mediante la reacción en cadena de la polimerasa (PCR) y se procedió a la secuenciación nucleotídica, luego de lo cual se analizó la presencia de mutaciones asociadas a resistencia a ARVS previamente reportadas. En la población tratada con ARVs, se encontraron mutaciones asociadas a resistencia en el 90,3 por ciento de las muestras. La prevalencia de mutaciones asociadas a resistencia a AZT, 3TC, los inhibidores no nucleosídicos de la RT (NNRTIs), indinavir, ritonavir y nelfinavir superó el 40 por ciento, siendo las sustituciones más prevalentes: M46I/L, V82A/F/S/T y L90M en la PR; y M41L, D67N, K70R, K103N, Y181C/I, M184I/V, G190A/S y T215F/Y en la RT. La presencia de variantes resistentes a miembros de múltiples familiars de drogas fue superior al 20 por ciento... (TRUNCADO) (AU)
Asunto(s)
Humanos , Masculino , Femenino , VIH , Resistencia a Medicamentos , Antivirales , Proteasa del VIH , ADN Polimerasa Dirigida por ARN , Seropositividad para VIH , Mutación/inmunología , Reacción en Cadena de la Polimerasa , Nucleósidos , Ensayos Clínicos como Asunto , Estudios de Seguimiento , ArgentinaRESUMEN
La instauración de tratamientos antirretrovirales de alta eficacia (HAARTs) ha llevado a la disminución de la morbimortalidad asociada a HIV-1/SIDA en países industrializados. Sin embargo, uno de los principales factores asociados a la falla terapéutica es la selección de variantes de HIV-1 resistentes a las drogas antirretrovirales (ARVs). Dichas variantes virales se caracterizan por presentar mutaciones en las enzimas virales que son los blancos de la acción de los fármacos: la proteasa viral (PR) y la transcriptasa reversa (RT). Las variantes del HIV-1 resitente a ARVs seleccionadas en pacientes tratados no sólo se asocian al fracaso terapéutico en dichos individuos, sino que también pueden ser transmitidas a la población no infectada. De esta forma, individuos infectados con HIV-1 que nunca han sido expuestos a drogas ARVs (naïve) pueden albergar virus resistentes, lo que llevaría a la falla terapéutica al iniciarse el tratamiento. El objetivo del presente trabajo fue el de caracterizar los perfiles de resistencia en pacientes HIV-1 seropositivos con falla terapéutica. Asimismo, con el fin de estimar la tasa de transmisión de variantes resistentes a ARVs, se estudiaron los perfiles de resistencia en individuos infectados con HIV-1 que no habían recibido tratamiento. Se estudiaron 399 pacientes HIV-1 seropositivos con falla terapéutica y 94 individuos naive infectados con HIV-1. Las muestras de plasma fueron recolectadas entre 1997 y 2001 de individuos de la Cdad. de Buenos Aires y sus alrededores. Se amplificaron las regiones de la PR y RT del HIV-1 mediante la reacción en cadena de la polimerasa (PCR) y se procedió a la secuenciación nucleotídica, luego de lo cual se analizó la presencia de mutaciones asociadas a resistencia a ARVS previamente reportadas. En la población tratada con ARVs, se encontraron mutaciones asociadas a resistencia en el 90,3 por ciento de las muestras. La prevalencia de mutaciones asociadas a resistencia a AZT, 3TC, los inhibidores no nucleosídicos de la RT (NNRTIs), indinavir, ritonavir y nelfinavir superó el 40 por ciento, siendo las sustituciones más prevalentes: M46I/L, V82A/F/S/T y L90M en la PR; y M41L, D67N, K70R, K103N, Y181C/I, M184I/V, G190A/S y T215F/Y en la RT. La presencia de variantes resistentes a miembros de múltiples familiars de drogas fue superior al 20 por ciento... (TRUNCADO)