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Protein kinases are highly tractable targets for drug discovery. However, the biological function and therapeutic potential of the majority of the 500+ human protein kinases remains unknown. We have developed physical and virtual collections of small molecule inhibitors, which we call chemogenomic sets, that are designed to inhibit the catalytic function of almost half the human protein kinases. In this manuscript we share our progress towards generation of a comprehensive kinase chemogenomic set (KCGS), release kinome profiling data of a large inhibitor set (Published Kinase Inhibitor Set 2 (PKIS2)), and outline a process through which the community can openly collaborate to create a KCGS that probes the full complement of human protein kinases.
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Bases de Datos Farmacéuticas , Inhibidores de Proteínas Quinasas/farmacología , Bibliotecas de Moléculas Pequeñas/farmacología , Descubrimiento de Drogas/métodos , Genómica/métodos , Humanos , Inhibidores de Proteínas Quinasas/química , Bibliotecas de Moléculas Pequeñas/química , Relación Estructura-ActividadRESUMEN
Chemical probes are required for preclinical target validation to interrogate novel biological targets and pathways. Selective inhibitors of the CREB binding protein (CREBBP)/EP300 bromodomains are required to facilitate the elucidation of biology associated with these important epigenetic targets. Medicinal chemistry optimization that paid particular attention to physiochemical properties delivered chemical probes with desirable potency, selectivity, and permeability attributes. An important feature of the optimization process was the successful application of rational structure-based drug design to address bromodomain selectivity issues (particularly against the structurally related BRD4 protein).
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Proteína de Unión a CREB/antagonistas & inhibidores , Diseño de Fármacos , Proteína p300 Asociada a E1A/antagonistas & inhibidores , Morfolinas/farmacología , Proteína de Unión a CREB/metabolismo , Química Farmacéutica , Relación Dosis-Respuesta a Droga , Proteína p300 Asociada a E1A/metabolismo , Humanos , Estructura Molecular , Morfolinas/síntesis química , Morfolinas/química , Relación Estructura-ActividadRESUMEN
Background University students' substance abuse and risky sex contribute to sexually transmitted diseases (STDs). Purpose We develop and empirically test a formative theoretical model of sexual temptation involving substance abuse (cigarettes, alcohol, and marijuana), safe sexual behavior (use of condom/barrier for oral and vaginal intercourse), risky sexual behavior (unprotected sex and multiple sexual partners), and STDs: gonorrhea, HIV, and genital herpes. We simultaneously explore these constructs, controlling membership in social groups (fraternity/sorority, varsity athlete, and club sports) and perceived norm of substance abuse. Methods A total of 687 American university students completed the National College Health Assessment (NCHA). We use structural equation modeling (SEM) to test the goodness of fit between our formative theoretical model and actual data. Results Results reveal the following discoveries: membership in campus social groups is positively associated with STDs, whereas perceived norm of peer substance abuse is negatively related to STDs. Under the influence of substance abuse, we test three outcomes of sexual temptation as related to STDs. Those who have no sex do not contract STDs. For those who fall into temptation and have sex, substance abuse is more strongly related to risky sex which leads to STDs than safe sex which does not. Those engaging in risky sex have significantly higher cognitive impairment than those practicing safe sex. Conclusions Substance abuse contributes to STDs through risky sex only. Those having risky sex suffer higher cognitive impairment than those practicing safe sex. We provide novel implications to policy makers, practitioners, and researchers.
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Small molecules that correct the folding defects and enhance surface localization of the F508del mutation in the Cystic Fibrosis Transmembrane conductance Regulator (CFTR) comprise an important therapeutic strategy for cystic fibrosis lung disease. However, compounds that rescue the F508del mutant protein to wild type (WT) levels have not been identified. In this report, we consider obstacles to obtaining robust and therapeutically relevant levels of F508del CFTR. For example, markedly diminished steady state amounts of F508del CFTR compared to WT CFTR are present in recombinant bronchial epithelial cell lines, even when much higher levels of mutant transcript are present. In human primary airway cells, the paucity of Band B F508del is even more pronounced, although F508del and WT mRNA concentrations are comparable. Therefore, to augment levels of "repairable" F508del CFTR and identify small molecules that then correct this pool, we developed compound library screening protocols based on automated protein detection. First, cell-based imaging measurements were used to semi-quantitatively estimate distribution of F508del CFTR by high content analysis of two-dimensional images. We evaluated ~2,000 known bioactive compounds from the NIH Roadmap Molecular Libraries Small Molecule Repository in a pilot screen and identified agents that increase the F508del protein pool. Second, we analyzed ~10,000 compounds representing diverse chemical scaffolds for effects on total CFTR expression using a multi-plate fluorescence protocol and describe compounds that promote F508del maturation. Together, our findings demonstrate proof of principle that agents identified in this fashion can augment the level of endoplasmic reticulum (ER) resident "Band B" F508del CFTR suitable for pharmacologic correction. As further evidence in support of this strategy, PYR-41-a compound that inhibits the E1 ubiquitin activating enzyme-was shown to synergistically enhance F508del rescue by C18, a small molecule corrector. Our combined results indicate that increasing the levels of ER-localized CFTR available for repair provides a novel route to correct F508del CFTR.
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Regulador de Conductancia de Transmembrana de Fibrosis Quística/metabolismo , Fibrosis Quística/patología , Retículo Endoplásmico/metabolismo , Bibliotecas de Moléculas Pequeñas/química , Alelos , Benzoatos/química , Benzoatos/farmacología , Células Cultivadas , Fibrosis Quística/genética , Regulador de Conductancia de Transmembrana de Fibrosis Quística/química , Regulador de Conductancia de Transmembrana de Fibrosis Quística/genética , Furanos/química , Furanos/farmacología , Eliminación de Gen , Células HEK293 , Células HeLa , Ensayos Analíticos de Alto Rendimiento , Humanos , Ácidos Hidroxámicos/química , Ácidos Hidroxámicos/farmacología , Microscopía Fluorescente , Pliegue de Proteína , Estructura Terciaria de Proteína , Pirazoles/química , Pirazoles/farmacología , ARN Mensajero/metabolismo , Bibliotecas de Moléculas Pequeñas/farmacología , Ubiquitinación/efectos de los fármacos , VorinostatRESUMEN
Bromodomains are involved in transcriptional regulation through the recognition of acetyl lysine modifications on diverse proteins. Selective pharmacological modulators of bromodomains are lacking, although the largely hydrophobic nature of the pocket makes these modules attractive targets for small-molecule inhibitors. This work describes the structure-based design of a highly selective inhibitor of the CREB binding protein (CBP) bromodomain and its use in cell-based transcriptional profiling experiments. The inhibitor downregulated a number of inflammatory genes in macrophages that were not affected by a selective BET bromodomain inhibitor. In addition, the CBP bromodomain inhibitor modulated the mRNA level of the regulator of G-protein signaling 4 (RGS4) gene in neurons, suggesting a potential therapeutic opportunity for CBP inhibitors in the treatment of neurological disorders.
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Proteína de Unión a CREB/antagonistas & inhibidores , Diseño de Fármacos , Bibliotecas de Moléculas Pequeñas/química , Proteína de Unión a CREB/genética , Transferencia Resonante de Energía de Fluorescencia , Regulación de la Expresión Génica/efectos de los fármacos , Humanos , Estructura Terciaria de Proteína , Proteínas RGS/genética , Bibliotecas de Moléculas Pequeñas/farmacología , TranscriptomaRESUMEN
Anisole and fluoroanisoles display distinct conformational preferences, as evident from a survey of their crystal structures. In addition to altering the free ligand conformation, various degrees of fluorination have a strong impact on physicochemical and pharmacokinetic properties. Analysis of anisole and fluoroanisole matched molecular pairs in the Pfizer corporate database reveals interesting trends: 1) PhOCF3 increases log D by ~1 log unit over PhOCH3 compounds; 2) PhOCF3 shows lower passive permeability despite its higher lipophilicity; and 3) PhOCF3 does not appreciably improve metabolic stability over PhOCH3 . Emerging from the investigation, difluoroanisole (PhOCF2 H) strikes a better balance of properties with noticeable advantages of log D and transcellular permeability over PhOCF3 . Synthetic assessment illustrates that the routes to access difluoroanisoles are often more straightforward than those for trifluoroanisoles. Whereas replacing PhOCH3 with PhOCF3 is a common tactic to optimize ADME properties, our analysis suggests PhOCF2 H may be a more attractive alternative, and greater exploitation of this motif is recommended.
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Anisoles/química , Diseño de Fármacos , Flúor/química , Animales , Anisoles/metabolismo , Anisoles/farmacocinética , Línea Celular , Perros , Flúor/metabolismo , Flúor/farmacocinética , Halogenación , Humanos , Ligandos , Microsomas Hepáticos/metabolismo , PermeabilidadRESUMEN
The Protein Data Bank is the most comprehensive source of experimental macromolecular structures. It can, however, be difficult at times to locate relevant structures with the Protein Data Bank search interface. This is particularly true when searching for complexes containing specific interactions between protein and ligand atoms. Moreover, searching within a family of proteins can be tedious. For example, one cannot search for some conserved residue as residue numbers vary across structures. We describe herein three databases, Protein Relational Database, Kinase Knowledge Base, and Matrix Metalloproteinase Knowledge Base, containing protein structures from the Protein Data Bank. In Protein Relational Database, atom-atom distances between protein and ligand have been precalculated allowing for millisecond retrieval based on atom identity and distance constraints. Ring centroids, centroid-centroid and centroid-atom distances and angles have also been included permitting queries for pi-stacking interactions and other structural motifs involving rings. Other geometric features can be searched through the inclusion of residue pair and triplet distances. In Kinase Knowledge Base and Matrix Metalloproteinase Knowledge Base, the catalytic domains have been aligned into common residue numbering schemes. Thus, by searching across Protein Relational Database and Kinase Knowledge Base, one can easily retrieve structures wherein, for example, a ligand of interest is making contact with the gatekeeper residue.
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Bases de Datos de Proteínas , Diseño de Fármacos , Bases del Conocimiento , Metaloproteinasas de la Matriz/química , Proteínas Quinasas/químicaRESUMEN
High throughput microsomal stability assays have been widely implemented in drug discovery and many companies have accumulated experimental measurements for thousands of compounds. Such datasets have been used to develop in silico models to predict metabolic stability and guide the selection of promising candidates for synthesis. This approach has proven most effective when selecting compounds from proposed virtual libraries prior to synthesis. However, these models are not easily interpretable at the structural level, and thus provide little insight to guide traditional synthetic efforts. We have developed global classification models of rat, mouse and human liver microsomal stability using in-house data. These models were built with FCFP_6 fingerprints using a Naïve Bayesian classifier within Pipeline Pilot. The test sets were correctly classified as stable or unstable with satisfying accuracies of 78, 77 and 75% for rat, human and mouse models, respectively. The prediction confidence was assigned using the Bayesian score to assess the applicability of the models. Using the resulting models, we developed a novel data mining strategy to identify structural features associated with good and bad microsomal stability. We also used this approach to identify structural features which are good for one species but bad for another. With these findings, the structure-metabolism relationships are likely to be understood faster and earlier in drug discovery.
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Microsomas Hepáticos/metabolismo , Preparaciones Farmacéuticas/química , Preparaciones Farmacéuticas/metabolismo , Relación Estructura-Actividad Cuantitativa , Animales , Descubrimiento de Drogas , Monitoreo del Ambiente , Femenino , Humanos , Masculino , Ratones , Estructura Molecular , RatasRESUMEN
A novel algorithm for the connecting of fragment molecules is presented and validated for a number of test systems. Within the CONFIRM (Connecting Fragments Found in Receptor Molecules) approach a pre-prepared library of bridges is searched to extract those which match a search criterion derived from known experimental or computational binding information about fragment molecules within a target binding site. The resulting bridge 'hits' are then connected, in an automated fashion, to the fragments and docked into the target receptor. Docking poses are assessed in terms of root-mean-squared deviation from the known positions of the fragment molecules, as well as docking score should known inhibitors be available. The creation of the bridge library, the full details and novelty of the CONFIRM algorithm, and the general applicability of this approach within the field of fragment-based de novo drug design are discussed.
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Algoritmos , Diseño de Fármacos , Modelos Moleculares , Proteínas/química , Receptores de Superficie Celular/química , Sitios de Unión , Bases de Datos Factuales , Humanos , Ligandos , Conformación Molecular , Estructura Molecular , Unión Proteica , Isoformas de Proteínas/química , Isoformas de Proteínas/metabolismo , Proteínas/metabolismo , Receptores de Superficie Celular/metabolismo , Receptores de Ácido Retinoico/química , Receptores de Ácido Retinoico/metabolismo , Estreptavidina/químicaRESUMEN
Assessment of risk posed by an environmental contaminant to an aquatic community requires estimation of both its magnitude of occurrence (exposure) and its ability to cause harm (effects). Our ability to estimate effects is often hindered by limited toxicological information. As a result, resource managers and environmental regulators are often faced with the need to extrapolate across taxonomic groups in order to protect the more sensitive members of the aquatic community. The goals of this effort were to 1) compile and organize an extensive body of acute toxicity data, 2) characterize the distribution of toxicant sensitivity across taxa and species, and 3) evaluate the utility of toxicity extrapolation methods based upon sensitivity relations among species and chemicals. Although the analysis encompassed a wide range of toxicants and species, pesticides and freshwater fish and invertebrates were emphasized as a reflection of available data. Although it is obviously desirable to have high-quality acute toxicity values for as many species as possible, the results of this effort allow for better use of available information for predicting the sensitivity of untested species to environmental contaminants. A software program entitled "Ecological Risk Analysis" (ERA) was developed that predicts toxicity values for sensitive members of the aquatic community using species sensitivity distributions. Of several methods evaluated, the ERA program used with minimum data sets comprising acute toxicity values for rainbow trout, bluegill, daphnia, and mysids provided the most satisfactory predictions with the least amount of data. However, if predictions must be made using data for a single species, the most satisfactory results were obtained with extrapolation factors developed for rainbow trout (0.412), bluegill (0.331), or scud (0.041). Although many specific exceptions occur, our results also support the conventional wisdom that invertebrates are generally more sensitive to contaminants than fish are.
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Peces/fisiología , Invertebrados/fisiología , Pruebas de Toxicidad/estadística & datos numéricos , Contaminantes Químicos del Agua/análisis , Contaminantes Químicos del Agua/toxicidad , Animales , Inhibidores de la Colinesterasa/toxicidad , Bases de Datos Factuales , Dosificación Letal Mediana , Medición de Riesgo , Programas Informáticos , Especificidad de la EspecieRESUMEN
The fate and toxicity of a polyethoxylated tallowamine (POEA) surfactant system, MON 0818, was evaluated in water-sediment microcosms during a 4-d laboratory study. A surfactant solution of 8 mg l(-1) nominal concentration was added to each of nine 72-l aquaria with or without a 3-cm layer of one of two natural sediments (total organic carbon (TOC) 1.5% or 3.0%). Control well water was added to each of nine additional 72-l aquaria with or without sediment. Water samples were collected from the microcosms after 2, 6, 24, 48, 72, and 96 h of aging to conduct 48-h toxicity tests with Daphnia magna and to determine surfactant concentrations. Elevated mortality of D. magna (43-83%) was observed in overlying water sampled from water-only microcosms throughout the 96-h aging period, whereas elevated mortality (23-97%) was only observed in overlying water sampled from water-sediment microcosms during the first 24h of aging. Measured concentrations of MON 0818 in water-only microcosms remained relatively constant (4-6 mg l(-1)) during the 96-h period, whereas the concentrations in overlying water from microcosms containing either of the two types of sediment dissipated rapidly, with half-lives of 13 h in the 3.0% TOC sediment and 18 h in the 1.5% TOC sediment. Both toxicity and the concentration of MON 0818 in overlying water decreased more rapidly in microcosms containing sediment with the higher percent TOC and clay and with a higher microbial biomass. Mortality of D. magna was significantly correlated with surfactant concentrations in the overlying water. These results indicate that the toxicity of the POEA surfactant in water rapidly declines in the presence of sediment due to a reduction in the surfactant concentration in the overlying water above the sediment.
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Daphnia/efectos de los fármacos , Grasas/toxicidad , Agua Dulce/análisis , Sedimentos Geológicos/química , Polietilenglicoles/toxicidad , Tensoactivos/toxicidad , Contaminantes Químicos del Agua/toxicidad , Animales , Daphnia/crecimiento & desarrollo , Pruebas de ToxicidadRESUMEN
Apyrases have been suggested to play important roles in plant nutrition, photomorphogenesis, and nodulation. To help trace the evolution of these genes in the legumes--and possibly, the acquisition of new functions for nodulation--apyrase-containing BACs were sequenced from three legume genomes. Genomic sequences from Medicago truncatula, Glycine max and Lotus japonicus were compared to one another and to corresponding regions in Arabidopsis thaliana. A phylogenetic analysis of apyrase homologs from these regions and sequences from other legume species, as well as other plant families, identified a potentially legume-specific clade that contains a well-characterized soybean ( G. soja) apyrase, Gs52, as well as homologs from Dolichos, Lotus, Medicago and Pisum. Sister clades contain homologs from members of Brassicaceae, Solanaceae, Poaceae and Fabaceae. Comparisons of rates of change at synonymous and nonsynonymous sites in the Gs52 and sister clades show rapid evolution in the potentially legume-specific Gs52 clade. The genomic organization of the apyrase-containing BACs shows evidence of gene duplication, genomic rearrangement, and gene conversion among Gs52 homologs. Taken together, these results suggest a scenario of local apyrase gene duplication in an ancestor of the legumes, followed by functional diversification and increased rates of change in the new genes, and further duplications in the Galegae (which include the genera Medicago and Pisum). The study also provides a detailed comparison of genomic regions between two model genomes which are now being sequenced ( M. truncatulaand L. japonicus), and a genome from an economically important legume species ( G. max).
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Apirasa/genética , Evolución Molecular , Fabaceae/genética , Sintenía , Cromosomas Artificiales Bacterianos , Fabaceae/enzimología , Duplicación de Gen , Variación Genética , Genoma , Modelos Genéticos , Filogenia , Homología de SecuenciaRESUMEN
The predictions of a class of phenomenological trap models of supercooled liquids are tested via computer simulation of a model glass-forming liquid. It is found that a model with a Gaussian distribution of trap energies provides a good description of the landscape dynamics, even at temperatures above T(c), the critical temperature of mode-coupling theory. A scenario is discussed whereby deep traps are composed of collections of inherent structures above T(c) and single inherent structures below T(c). Deviations from the simple Gaussian trap picture are quantified and discussed.
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En este estudio se establecieron los niveles de mercurio (Hg) en cabello de individuos profesionalmente expuestos en el área odontológica y se contrastaron con las concentraciones mercuriales de un grupo control no expuesto ocupacionalmente al Hg. Las muestras de cabello fueron recolectadas de 15 individuos expuestos y 15 no expuestos, residentes de la ciudad de Maracaibo y zonas circunvencians. Los individuos fueron interrogados a través de una encuesta en relación a datos personales, dirección de habitación, ocupación, alimentación, hábitos, salud, tratamiento médicos, número de restauraciones de amalgama, tratamientos cosméticos en el cabello, etc. Se tomaron 2 cm de cabello distal en la zona occipital del cuero cabelludo de cada individuos, las cuales se almacenaron en bolsas plásticas de cierre hermético a 4§C. Previo al análisis espectrométrico, las muestras se levaron y se desmineralizaron usando microondas. La determinacion exacta, precisa y libre de interferencias de Hg se realizó empleando la espectrometría de absorción atómica por vapor frío. La concentración media de Hg (+- 1 DE obtenida para el grupo expuesto (E) fue 2,07 +- 2,23 (ca. intervalo experimental: 0,13 - 7,91 mg Hg/g) y para el grupo control (C) fue 2,65 +- 2,06 (ca. intervalo experimental): 0,75 - 6,75 mg Hg/g). No se observaron diferencias estadísticamente significativas (p>0,05) entre los niveles de mercurio para los grupos E y C; por lo tanto, el vapor de Hg absorbido en la clínica dental no influyó en el nivel de Hg en cabello del grupo expuesto, debido posiblemente al uso de sistemas de aire acondicionado y buena ventilación que presentaron las clínicas. Los resultados obtenidos en este estudio revelaron que el manejo y uso apropiado de la amalgama dental no altera el nivel de mercurio presente en el cabello del personal que labora en la clínica dental
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Humanos , Adolescente , Adulto , Femenino , Persona de Mediana Edad , Amalgama Dental , Personal de Odontología , Exposición Profesional/estadística & datos numéricos , Cabello , Mercurio , Clínicas Odontológicas/normas , Recolección de Datos , Equipo Dental , Espectrofotometría Atómica/métodos , Encuestas Nutricionales , Enfermedades Profesionales , Restauración Dental Permanente/estadística & datos numéricos , Interpretación Estadística de Datos , Venezuela , Ventilación/métodosRESUMEN
We are building a framework physical infrastructure across the soybean genome by using SSR (simple sequence repeat) and RFLP (restriction fragment length polymorphism) markers to identify BACs (bacterial artificial chromosomes) from two soybean BAC libraries. The libraries were prepared from two genotypes, each digested with a different restriction enzyme. The BACs identified by each marker were grouped into contigs. We have obtained BAC- end sequence from BACs within each contig. The sequences were analyzed by the University of Minnesota Center for Computational Genomics and Bioinformatics using BLAST algorithms to search nucleotide and protein databases. The SSR-identified BACs had a higher percentage of significant BLAST hits than did the RFLP-identified BACs. This difference was due to a higher percentage of hits to repetitive-type sequences for the SSR-identified BACs that was offset in part, however, by a somewhat larger proportion of RFLP-identified significant hits with similarity to experimentally defined genes and soybean ESTs (expressed sequence tags). These genes represented a wide range of metabolic functions. In these analyses, only repetitive sequences from SSR-identified contigs appeared to be clustered. The BAC-end sequences also allowed us to identify microsynteny between soybean and the model plants Arabidopsis thaliana and Medicago truncatula. This map-based approach to genome sampling provides a means of assaying soybean genome structure and organization.
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Cromosomas Artificiales Bacterianos , Marcadores Genéticos , Glycine max/genética , Polimorfismo Genético , Polimorfismo de Longitud del Fragmento de Restricción , Algoritmos , Arabidopsis/genética , Mapeo Contig , Bases de Datos como Asunto , Etiquetas de Secuencia Expresada , Biblioteca de Genes , Genotipo , Medicago/genética , Modelos Genéticos , Datos de Secuencia Molecular , Análisis de Secuencia de ADN , Programas InformáticosAsunto(s)
Trasplante de Riñón , Donadores Vivos , Nefrectomía/métodos , Páncreas , Pancreatectomía/métodos , Humanos , Masculino , Persona de Mediana EdadRESUMEN
A fully microscopic molecular hydrodynamic theory for the two-dimensional (fifth-order) Raman spectrum of an atomic liquid (Xe) is presented. The spectrum is obtained from a simple mode-coupling theory by projecting the dynamics onto bilinear pairs of fluctuating density variables. Good agreement is obtained in comparison with recently reported molecular dynamics simulation results. The microscopic theory provides an understanding of the timescales and molecular motions that govern the two-dimensional signal. Predictions are made for the behavior of the spectrum as a function of temperature and density. The theory shows that novel signatures in the two-dimensional Raman spectrum of supercritical and supercooled liquids are expected.
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BACKGROUND: Nonhuman primates (NHPs) have been widely used in different porcine xenograft procedures inevitably resulting in exposure to porcine endogenous retrovirus (PERV). Surveillance for PERV infection in these NHPs may provide information on the risks of cross-species transmission of PERV, particularly for recipients of vascularized organ xenografts for whom data from human clinical trials is unavailable. METHODS: We tested 21 Old World and 2 New World primates exposed to a variety of porcine xenografts for evidence of PERV infection. These NHPs included six baboon recipients of pig hearts, six bonnet macaque recipients of transgenic pig skin grafts, and nine rhesus macaque and two capuchin recipients of encapsulated pig islet cells. Serologic screening for PERV antibody was done by a validated Western blot assay, and molecular detection of PERV sequences in peripheral blood mononuclear cells (PBMCs) and plasma was performed using sensitive polymerase chain reaction and reverse transcriptase-polymerase chain reaction assays, respectively. Spleen and lymph node tissues available from six bonnet macaques and three rhesus macaques were also tested for PERV sequences. RESULTS: All plasma samples were negative for PERV RNA suggesting the absence of viremia in these xenografted animals. Similarly, PERV sequences were not detectable in any PBMC and tissue samples, arguing for the lack of latent infection of these compartments. In addition, all plasma samples were negative for PERV antibodies. CONCLUSION: These data suggest the absence of PERV infection in all 23 NHPs despite exposure to vascularized porcine organs or tissue xenografts and the use of immunosuppressive therapies in some animals. These findings suggest that PERV is not easily transmitted to these NHP species through these types of xenografts.
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Cebidae/virología , Trasplante de Células/efectos adversos , Cercopithecidae/virología , Trasplante de Órganos/efectos adversos , Infecciones por Retroviridae/transmisión , Enfermedades de los Porcinos/transmisión , Trasplante Heterólogo/efectos adversos , Animales , Cebus , Quimera , Islotes Pancreáticos/citología , Macaca , Papio , ARN Viral/análisis , Retroviridae/genética , Retroviridae/inmunología , Trasplante de Piel/efectos adversos , Porcinos/genética , Porcinos/virologíaRESUMEN
Four putative apyrase genes were identified from the model legume Medicago truncatula. Two of the genes identified from M. truncatula (Mtapy1 and Mtapy4) are expressed in roots and are inducible within 3 h after inoculation with Sinorhizobium meliloti. The level of mRNA expression of the other two putative apyrases, Mtapy2 and Mtapy3, was unaffected by rhizobial inoculation. Screening of a bacterial artificial chromosome library of M. truncatula genomic DNA showed that Mtapy1, Mtapy3, and Mtapy4 are present on a single bacterial artificial chromosome clone. This apyrase cluster was mapped to linkage group seven. A syntenic region on soybean linkage group J was found to contain at least two apyrase genes. Screening of nodulation deficient mutants of M. truncatula revealed that two such mutants do not express apyrases to any detectable level. The data suggest a role for apyrases early in the nodulation response before the involvement of root cortical cell division leading to the nodule structure.
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Apirasa/genética , Regulación de la Expresión Génica de las Plantas , Medicago sativa/enzimología , Medicago sativa/genética , Sinorhizobium/fisiología , Transcripción Genética , Secuencia de Aminoácidos , Apirasa/biosíntesis , Apirasa/química , Secuencia Conservada , Inducción Enzimática , Regulación Enzimológica de la Expresión Génica , Humanos , Isoenzimas/biosíntesis , Isoenzimas/química , Isoenzimas/genética , Medicago sativa/microbiología , Datos de Secuencia Molecular , Raíces de Plantas/enzimología , Plantas/enzimología , Plantas/genética , Reacción en Cadena de la Polimerasa , ARN Mensajero/genética , Alineación de Secuencia , Homología de Secuencia de Aminoácido , Factores de TiempoRESUMEN
BACKGROUND: The most common surgical complication after a kidney transplant is likely related to the wound. The purpose of this analysis was to determine the incidence of, and risk factors for, wound complications (e.g., infections, hernias) in kidney recipients and to assess whether newer immunosuppressive drugs increase the risk for such complications. METHODS: Between January 1, 1984 and September 30, 1998, we performed 2013 adult kidney transplants. Of these 2013 recipients, 97 (4.8%) developed either a superficial or a deep wound infection. Additionally, 73 (3.6%) recipients developed either a fascial dehiscence or a hernia of the wound. We used univariate and multivariate techniques to determine significant risk factors and outcomes. RESULTS: Mean time to development of a superficial infection (defined as located above the fascia) was 11.9 days posttransplant; to development of a deep infection (defined as located below the fascia), 39.2 days; and to development of a hernia or fascial dehiscence, 12.8 months. By multivariate analysis, the most significant risk factor for a superficial or deep wound infection was obesity (defined as body mass index>30 kg/m2) (RR=4.4, P=0.0001). Other significant risk factors were a urine leak posttransplant, any reoperation through the transplant incision, diabetes, and the use of mycophenolate mofetil (MMF) (vs. azathioprine) for maintenance immunosuppression (RR=2.43, P=0.0001). Significant risk factors for a hernia or fascial dehiscence were any reoperation through the transplant incision, increased recipient age, obesity, and the use of MMF (vs. azathioprine) for maintenance immunosuppression (RR=3.54, P=0.0004). Use of antibody induction and treatment for acute rejection were not significant risk factors for either infections or hernias. Death-censored graft survival was lower in recipients who developed a wound infection (vs. those who did not); it was not lower in recipients who developed an incisional hernia or facial dehiscence (vs. those who did not). CONCLUSIONS: Despite immunosuppression including chronic steroids, the incidence of wound infections, incisional hernias, and fascial dehiscence is low in kidney recipients. As with other types of surgery, the main risk factors for postoperative complications are obesity, reoperation, and increased age. However, in kidney recipients, use of MMF (vs. azathioprine) is an additional risk factor -one that potentially could be altered, especially in high-risk recipients.