RESUMEN
Low-sodium and high-potassium diets have been recommended as an adjunct to prevention and treatment of hypertension. Analysis of these nutrients in 24-h urine has been considered the reference method to estimate daily intake of these minerals. However, 24-h urine collection is difficult in epidemiological studies, since urine must be collected and stored in job environments. Therefore, strategies for shorter durations of urine collection at home have been proposed. We have previously reported that collecting urine during a 12-h period (overnight) is more feasible and that creatinine clearance correlated strongly with that detected in 24-h samples. In the present study, we collected urine for 24 h divided into two 12-h periods (from 7:00 am to 7:00 pm and from 7:00 pm to 7:00 am next day). A sample of 109 apparently healthy volunteers aged 30 to 74 years of both genders working in a University institution was investigated. Subjects with previous myocardial infarction, stroke, renal insufficiency, and pregnant women were not included. Significant (P < 0.001) Spearman correlation coefficients (r s) were found between the total amount of sodium and potassium excreted in the urine collected at night and in the 24-h period (r s = 0.76 and 0.74, respectively). Additionally, the 12-h sodium and potassium excretions (means ± SD, 95% confidence interval) corresponded to 47.3 ± 11.2%, 95%CI = 45.3-49.3, and 39.3 ± 4.6%, 95%CI = 37.3-41.3, respectively, of the 24-h excretion of these ions. Therefore, these findings support the assumption that 12-h urine collected at night can be used as a reliable tool to estimate 24-h intake/excretion of sodium and potassium.
Asunto(s)
Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Potasio/orina , Sodio/orina , Toma de Muestras de Orina/métodos , Estudios Transversales , Creatinina/orina , Potasio en la Dieta , Cloruro de Sodio Dietético , Factores de TiempoRESUMEN
Iron is an essential trace element for almost all organisms. However, an excess of this metal within cells can be deleterious on account of catalysis of cell-damaging hydroxyl radicals. Therefore, the concentration of iron within cells is tightly regulated and the primary control occurs by regulating its uptake. Under conditions of low iron availability, most fungi mobilize extracellular iron by excretion of low-molecular-mass ferric iron chelators, termed siderophores. Due to the potential impact of iron metabolism on fungal pathogenicity, a better insight into siderophore-mediated iron uptake is needed. In Aspergillus nidulans, siderophore biosynthesis and uptake are negatively regulated by the GATA-type transcription factor SREA. Hence, genes involved in siderophore biosynthesis and uptake are characterized by transcriptional induction under iron limitation in wild-type strain and de-repression in an sreA-deletion strain under conditions of sufficient iron supply. Such genes have been searched for using different strategies, e.g. differential mRNA display and expression analysis of candidate genes from various A. nidulans sequence databases. The identified genes presumably encode enzymes needed for siderophore biosynthesis, and transporters involved in siderophore uptake and/or excretion. The functional characterization of these genes will help to unravel the pathways involved in siderophore biosynthesis and uptake.
Asunto(s)
Aspergillus nidulans/genética , Regulación Fúngica de la Expresión Génica/genética , Regulón/genética , Sideróforos/metabolismo , Aspergillus nidulans/metabolismo , Proteínas Fúngicas/genética , Factores de Transcripción GATA , Genes Fúngicos , Proteínas Represoras/genética , Sideróforos/biosíntesisRESUMEN
Under conditions of low iron availability, most fungi excrete siderophores in order to mobilize extracellular iron. We show that lack of the GATA-type transcription factor SREA in Aspergillus nidulans not only leads to derepression of siderophore biosynthesis but also to deregulation of siderophore-bound iron uptake and ornithine esterase expression. Furthermore, SREA deficiency causes increased accumulation of ferricrocin, the siderophore responsible for intracellular iron storage. In sreA deletion strains, extracellular siderophore production is derepressed but still regulated negatively by iron availability, indicating the presence of an additional iron-regulatory mechanism. In contrast, iron affects ferricrocin accumulation in a positive way, suggesting a protective role for this siderophore in detoxification of intracellular iron excess. The harmfulness of deregulated iron uptake in this mutant is demonstrated by increased expression of genes encoding the antioxidative enzymes catalase CATB and the superoxide dismutases SODA and SODB. It is noteworthy that iron starvation was found to repress catB expression in wild-type (wt) and SREA-deficient strains, consistent with catB being subject to SREA-independent iron regulation. Differential display led to the identification of putative SREA target genes amcA and mirA. The deduced MIRA amino acid sequence displays significant similarity to recently characterized siderophore permeases of Saccharomyces cerevisiae. amcA encodes a putative mitochondrial carrier for the siderophore precursor ornithine, indicating cross-regulation of siderophore and ornithine metabolism.
Asunto(s)
Aspergillus nidulans/metabolismo , Proteínas Bacterianas/genética , Proteínas Portadoras/genética , Ferricromo/análogos & derivados , Proteínas Fúngicas/genética , Regulación Fúngica de la Expresión Génica , Proteínas de Transporte de Membrana/genética , Proteínas Mitocondriales/genética , Proteínas Represoras/metabolismo , Sideróforos/biosíntesis , Sideróforos/metabolismo , Secuencia de Aminoácidos , Aspergillus nidulans/genética , Aspergillus nidulans/crecimiento & desarrollo , Proteínas Bacterianas/química , Proteínas Bacterianas/metabolismo , Proteínas Portadoras/química , Proteínas Portadoras/metabolismo , Clonación Molecular , Esterasas/metabolismo , Ferricromo/metabolismo , Proteínas Fúngicas/química , Proteínas Fúngicas/metabolismo , Factores de Transcripción GATA , Hierro/metabolismo , Proteínas de Transporte de Membrana/química , Proteínas de Transporte de Membrana/metabolismo , Proteínas de Transporte de Membrana Mitocondrial , Proteínas Mitocondriales/química , Proteínas Mitocondriales/metabolismo , Datos de Secuencia Molecular , Ornitina/metabolismo , Proteínas Represoras/genética , Alineación de Secuencia , Análisis de Secuencia de ADN , Superóxido Dismutasa/genética , Superóxido Dismutasa/metabolismoRESUMEN
A highly inducible fungal promoter derived from the Penicillium chrysogenum endoxylanase (xylP) gene is described. Northern analysis and the use of a beta-glucuronidase (uidA) reporter gene strategy showed that xylP expression is transcriptionally regulated. Xylan and xylose are efficient inducers, whereas glucose strongly represses the promoter activity. Comparison of the same expression construct as a single copy at the niaD locus in P. chrysogenum and at the argB locus in Aspergillus nidulans demonstrated that the xylP promoter is regulated similarly in these two species but that the level of expression is about 80 times higher in the Aspergillus species. The xylP promoter was found to be 65-fold more efficient than the isopenicillin-N-synthetase (pcbC) promoter in Penicillium and 23-fold more efficient than the nitrate reductase (niaD) promoter in Aspergillus under induced conditions. Furthermore, the xylP promoter was used for controllable antisense RNA synthesis of the nre-encoded putative major nitrogen regulator of P. chrysogenum. This approach led to inducible downregulation of the steady-state mRNA level of nre and consequently to transcriptional repression of the genes responsible for nitrate assimilation. In addition, transcription of nreB, which encodes a negative-acting nitrogen regulatory GATA factor of Penicillium, was found to be subject to regulation by NRE. Our data are the first direct evidence that nre indeed encodes an activator in the nitrogen regulatory circuit in Penicillium and indicate that cross regulation of the controlling factors occurs.
Asunto(s)
Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Penicillium chrysogenum/enzimología , Regiones Promotoras Genéticas/genética , ARN sin Sentido/metabolismo , Factores de Transcripción/genética , Factores de Transcripción/metabolismo , Xilosidasas/genética , Aspergillus nidulans/enzimología , Aspergillus nidulans/genética , Secuencia de Bases , Regulación hacia Abajo , Endo-1,4-beta Xilanasas , Regulación Fúngica de la Expresión Génica , Genes Reporteros , Glucuronidasa/genética , Glucuronidasa/metabolismo , Datos de Secuencia Molecular , Penicillium chrysogenum/genética , ARN sin Sentido/genética , Análisis de Secuencia de ADN , Transcripción Genética , Xilosidasas/metabolismoRESUMEN
Human tear lipocalin (TL) is an unusual member of the lipocalin protein family, since it is known to bind a large variety of lipophilic ligands in vivo and acts as a cysteine proteinase inhibitor in vitro. It is suggested to function as a physiological protection factor by scavenging lipophilic potentially harmful compounds. Since protein-protein interaction or macromolecular complexation is a common feature of many lipocalins, we applied phage display technology to identify TL interacting proteins. By panning of a human prostate cDNA phagemid library against purified TL we isolated a thioredoxin (Trx) encoding phage clone. Biochemical analysis revealed that TL indeed interacts with Trx and is reduced by this redox protein. Reduction of the TL-specific disulfide bond is of functional relevance, since the reduced protein shows a nine-fold increase in ligand affinity when tested with retinoic acid as ligand.
Asunto(s)
Proteínas Portadoras/química , Tiorredoxinas/química , Secuencia de Aminoácidos , Secuencia de Bases , Proteínas Portadoras/genética , Clonación Molecular , ADN Complementario/aislamiento & purificación , Disulfuros/química , Electroforesis en Gel de Poliacrilamida , Proteínas del Ojo/química , Fluorescencia , Biblioteca de Genes , Humanos , Ligandos , Lipocalina 1 , Masculino , Datos de Secuencia Molecular , Oxidación-Reducción , Unión Proteica , Tiorredoxinas/genética , Tretinoina/químicaAsunto(s)
Angioplastia Coronaria con Balón , Enfermedad Coronaria/terapia , Adulto , Circulación Colateral/fisiología , Angiografía Coronaria , Enfermedad Coronaria/diagnóstico por imagen , Humanos , Masculino , Infarto del Miocardio/diagnóstico por imagen , Infarto del Miocardio/terapia , PronósticoRESUMEN
The stress on the articular surface of the hip joint is calculated for a group of joints with idiopathic osteoarthrosis of the hip and a healthy control group. The results indicate that compressive stress is of minor importance with respect to the etiology of idiopathic osteoarthrosis of the hip joint.