RESUMEN
Despite extensive studies suggesting increased susceptibility to HIV during the secretory phase of the menstrual cycle, the molecular mechanisms involved remain unclear. Our goal was to analyze transcriptomes of the endocervix and ectocervix during the proliferative and secretory phases using RNA sequencing to explore potential molecular signatures of susceptibility to HIV. We identified 202 differentially expressed genes (DEGs) between the proliferative and secretory phases of the cycle in the endocervix (adjusted p < 0.05). The biofunctions and pathways analysis of DEGs revealed that cellular assembly and epithelial barrier function in the proliferative phase and inflammatory response/cellular movement in the secretory phase were among the top biofunctions and pathways. The gene set enrichment analysis of ranked DEGs (score = log fold change/p value) in the endocervix and ectocervix revealed that (i) unstimulated/not activated immune cells gene sets positively correlated with the proliferative phase and negatively correlated with the secretory phase in both tissues, (ii) IFNγ and IFNα response gene sets positively correlated with the proliferative phase in the ectocervix, (iii) HIV restrictive Wnt/ß-catenin signaling pathway negatively correlated with the secretory phase in the endocervix. Our data show menstrual cycle phase-associated changes in both endocervix and ectocervix, which may modulate susceptibility to HIV.
Asunto(s)
Cuello del Útero/metabolismo , Fase Folicular/genética , Perfilación de la Expresión Génica , Fase Luteínica/genética , Transcriptoma , Biología Computacional/métodos , Endometrio/metabolismo , Femenino , Ontología de Genes , Redes Reguladoras de Genes , Humanos , Transducción de SeñalRESUMEN
Major depressive disorder in the elderly is associated with increased morbidity and reduced quality of life. This 10 week, placebo-controlled study investigated the efficacy and tolerability of extended-release bupropion (150-300 mg once daily) in depressed patients aged 65 years or older. The statistical assumptions necessary for the validity of the protocol-specified analysis of covariance were not met for the analysis of the primary outcome variable (Montgomery-Asberg Depression Rating Scale total score at Week 10, last observation carried forward). Alternative statistical methods used for the analysis of this variable demonstrated statistical significance. Statistically significant improvements were observed on the majority of secondary end points when compared with placebo, including the health outcome measures for motivation and energy, and life satisfaction and contentment. Adverse events were generally mild to moderate and similar between treatment groups. This study demonstrated that the extended-release bupropion is an effective, well-tolerated treatment for major depression in the elderly.
Asunto(s)
Antidepresivos de Segunda Generación/uso terapéutico , Bupropión/uso terapéutico , Trastorno Depresivo Mayor/tratamiento farmacológico , Inhibidores de Captación de Dopamina/uso terapéutico , Factores de Edad , Anciano , Anciano de 80 o más Años , Antidepresivos de Segunda Generación/efectos adversos , Australia , Bupropión/efectos adversos , Preparaciones de Acción Retardada , Trastorno Depresivo Mayor/diagnóstico , Trastorno Depresivo Mayor/psicología , Inhibidores de Captación de Dopamina/efectos adversos , Método Doble Ciego , Europa (Continente) , Femenino , Humanos , Masculino , Motivación , América del Norte , Satisfacción Personal , Efecto Placebo , Escalas de Valoración Psiquiátrica , Calidad de Vida , Sudáfrica , Factores de Tiempo , Resultado del TratamientoRESUMEN
The efficacy, safety and tolerability of bupropion XR and venlafaxine XR was assessed and compared with placebo in adult outpatients with major depressive disorder (MDD). Adults meeting DSM-IV criteria for MDD with a minimum Hamilton Depression Rating Scale (HAMD) 17-Item total score of > or =18 were randomized to eight weeks of double-blind treatment with either bupropion XR (150 mg/day), venlafaxine XR (75 mg/day) or placebo. At the end of the fourth week of treatment, a dosage increase to bupropion XR 300 mg/day or venlafaxine XR 150 mg/day was allowed if, in the opinion of the investigator, response was inadequate. The primary efficacy endpoint was mean change from baseline at week 8 in the Montgomery-Asberg Depression Rating Scale (MADRS) total score last observation carried forward (LOCF). Mean changes from baseline at week 8 (LOCF) in MADRS total score were statistically significant for bupropion XR and venlafaxine XR patients compared to the placebo group: -16.0 for bupropion XR (P = 0.006 vs placebo), -17.1 for venlafaxine XR (P < 0.001 vs placebo) and -13.5 for placebo. Secondary outcomes (including CGI-S, HAM-A, MEI, Q-LES-Q-SF, responder and remitter analyses) also improved significantly for both active treatment groups compared with placebo. The most frequently reported adverse events were dry mouth and insomnia for bupropion XR, and nausea, hyperhidrosis, fatigue, and insomnia for venlafaxine XR. In this double-blind, placebo-controlled trial, bupropion XR at doses up to 300 mg/day and venlafaxine XR at doses up to 150 mg/day demonstrated comparable antidepressant efficacy.
Asunto(s)
Antidepresivos de Segunda Generación/uso terapéutico , Bupropión/uso terapéutico , Ciclohexanoles/uso terapéutico , Trastorno Depresivo Mayor/tratamiento farmacológico , Adulto , Antidepresivos de Segunda Generación/administración & dosificación , Antidepresivos de Segunda Generación/efectos adversos , Bupropión/administración & dosificación , Bupropión/efectos adversos , Ciclohexanoles/administración & dosificación , Ciclohexanoles/efectos adversos , Preparaciones de Acción Retardada , Método Doble Ciego , Femenino , Humanos , Masculino , Persona de Mediana Edad , Escalas de Valoración Psiquiátrica , Psicometría , Clorhidrato de VenlafaxinaRESUMEN
BACKGROUND: ATP binding cassette (ABC) transporters are ubiquitously distributed transmembrane solute pumps that play a causative role in numerous diseases. Previous structures have defined the fold of the ABC and established the flexibility of its alpha-helical subdomain. But the nature of the mechanical changes that occur at each step of the chemical ATPase cycle have not been defined. RESULTS: Crystal structures were determined of the MJ1267 ABC from Methanococcus jannaschii in Mg-ADP-bound and nucleotide-free forms. Comparison of these structures reveals an induced-fit effect at the active site likely to be a consequence of nucleotide binding. In the Mg-ADP-bound structure, the loop following the Walker B moves toward the Walker A (P-loop) coupled to backbone conformational changes in the intervening "H-loop", which contains an invariant histidine. These changes affect the region believed to mediate intercassette interaction in the ABC transporter complex. Comparison of the Mg-ADP-bound structure of MJ1267 to the ATP-bound structure of HisP suggests that an outward rotation of the alpha-helical subdomain is coupled to the loss of a molecular contact between the gamma-phosphate of ATP and an invariant glutamine in a segment connecting this subdomain to the core of the cassette. CONCLUSIONS: The induced-fit effect and rotation of the alpha-helical subdomain may play a role in controlling the nucleotide-dependent change in cassette-cassette interaction affinity believed to represent the power-stroke of ABC transporters. Outward rotation of the alpha-helical subdomain also likely facilitates Mg-ADP release after hydrolysis. The MJ1267 structures therefore define features of the nucleotide-dependent conformational changes that drive transmembrane transport in ABC transporters.
Asunto(s)
Transportadoras de Casetes de Unión a ATP/química , Transportadoras de Casetes de Unión a ATP/metabolismo , Adenosina Trifosfato/metabolismo , Sistemas de Transporte de Aminoácidos Básicos , Proteínas Bacterianas , Secuencia de Aminoácidos , Sitios de Unión , Dominio Catalítico , Secuencia Conservada , Cristalografía por Rayos X , Magnesio/metabolismo , Proteínas de Transporte de Membrana/química , Proteínas de Transporte de Membrana/metabolismo , Methanococcus/química , Methanococcus/metabolismo , Modelos Moleculares , Datos de Secuencia Molecular , Fosfatos/metabolismo , Filogenia , Conformación ProteicaRESUMEN
The crystal structure of the MJ0796 ATP-binding cassette, a member of the o228/LolD transporter family, has been determined at 2.7-A resolution with MgADP bound at its active site. Comparing this structure with that of the ATP-bound form of the HisP ATP-binding cassette (Hung, L. W., Wang, I. X., Nikaido, K., Liu, P. Q., Ames, G. F., and Kim, S. H. (1998) Nature 396, 703-707) shows a 5-A withdrawal of a phylogenetically invariant glutamine residue from contact with the gamma-phosphate of ATP in the active site. This glutamine is located in a protein segment that links the rigid F(1)-type ATP-binding core of the enzyme to an ABC transporter-specific alpha-helical subdomain that moves substantially away from the active site in the MgADP-bound structure of MJ0796 compared with the ATP-bound structure of HisP. A similar conformational effect is observed in the MgADP-bound structure of MJ1267 (Karpowich, N., et al. (2001) Structure, in press), establishing the withdrawal of the glutamine and the coupled outward rotation of the alpha-helical subdomain as consistent consequences of gamma-phosphate release from the active site of the transporter. Considering this subdomain movement in the context of a leading model for the physiological dimer of cassettes present in ABC transporters indicates that it produces a modest mechanical change that is likely to play a role in facilitating nucleotide exchange out of the ATPase active site. Finally, it is noteworthy that one of the intersubunit packing interactions in the MJ0796 crystal involves antiparallel beta-type hydrogen bonding interactions between the outermost beta-strands in the two core beta-sheets, leading to their fusion into a single extended beta-sheet, a type of structural interaction that has been proposed to play a role in mediating the aggregation of beta-sheet-containing proteins.
Asunto(s)
Transportadoras de Casetes de Unión a ATP/química , Adenosina Trifosfato/metabolismo , Transportadoras de Casetes de Unión a ATP/metabolismo , Adenosina Difosfato/metabolismo , Secuencia de Aminoácidos , Sitios de Unión , Cristalografía por Rayos X , Modelos Moleculares , Datos de Secuencia Molecular , Filogenia , Conformación Proteica , Homología de Secuencia de AminoácidoRESUMEN
The most common disease-causing mutation in the cystic fibrosis transmembrane conductance regulator is a single amino acid deletion (DeltaF508) in the N-terminal cytosolic nucleotide-binding domain (NBD1). This mutation has previously been shown to be a temperature-sensitive folding mutation that alters the folding pathway but not the native state stability of the isolated domain (Qu, B.-H., and Thomas, P. J. (1996) J. Biol. Chem. 271, 7261-7264). Here we provide evidence that the molecular chaperone Hsc70 productively interacts with NBD1 to increase the folding yield of the domain and inhibit off-pathway associations leading to the formation of high molecular weight aggregates. Furthermore, we have sublocalized a region within NBD1 where Hsc70 binds. Notably, inhibition of NBD1 aggregation is not dependent upon the presence of Hsc70 in the early stages of folding, indicating that the chaperone may act on a folding intermediate. In the presence of K+ and Mg2+-ATP, conditions where Hsp70 binds substrate rapidly and can release it, Hsc70 is less effective at inhibiting NBD1 aggregation. Thus, the rate of release of unfolded substrate is an important factor in preventing aggregation and promoting folding of the domain. These results demonstrate that Hsc70 promotes the otherwise inefficient folding of DeltaF-NBD1 and provide insight into the mechanisms by which molecular chaperones assist proteins in folding.
Asunto(s)
Proteínas Portadoras/metabolismo , Regulador de Conductancia de Transmembrana de Fibrosis Quística/metabolismo , Proteínas HSP70 de Choque Térmico/metabolismo , Pliegue de Proteína , Adenosina Trifosfato/metabolismo , Animales , Sitios de Unión , Química Encefálica , Bovinos , Regulador de Conductancia de Transmembrana de Fibrosis Quística/química , Proteínas del Choque Térmico HSC70RESUMEN
Factor analysis of a questionnaire designed to measure a son's feeling of being understood by his father is reported. The questionnaire, which contained 12 paragraphs each purporting to be an aspect of the son's feeling of being understood by his father, was administered in small groups to a sample of 413 male college students along with three indices of son's level of general functioning--a self-concept scale, a somatic complaints inventory, and the degree of help-seeking behavior. Five interpretable factors were extracted from the data. The meaningfulness of the factors was substantiated by their significant correlations with certain of the other measures administered to the sample. Three of the factors were correlated positively (p less than .01) with Ss' self-concept, while two were correlated negatively (p less than .01) with the number of somatic complaints by the Ss'.