RESUMEN
AIMS: To evaluate a low-cost water quality test for at-scale drinking water safety estimation in rural India. METHODS AND RESULTS: Within a longitudinal study to characterize variability in household drinking water safety in rural Maharashtra, we piloted a low-cost presence-absence (LCPA) microbial test designed to be used by volunteer residents in rural areas. In comparing the LCPA results with standard laboratory methods for enumeration of Escherichia coli, we found that LCPA tests using modified mTec media were highly sensitive in detecting drinking water of moderate risk (88% of tests were positive at E. coli counts of 11-100 CFU per 100 ml) and high risk (96% of tests were positive at E. coli counts of 101 + CFU per 100 ml). The LCPA tests demonstrated low specificity for E. coli specifically, due to concurrent detection of Klebsiella: 38% of LCPA tests were positive even when E. coli was not detected in a 100 ml sample by membrane filtration, suggesting the test would be conservative in risk estimation. We also found that 47% of participants in rural villages in India were willing to conduct tests and return results after a brief training, with 45% of active participants sending their water testing results via short message service. CONCLUSIONS: Given their low cost (~US$0.50 as piloted) and open-source format, such tests may provide a compelling alternative to standard methods for rapid water quality assessments, especially in resource-limited settings. SIGNIFICANCE AND IMPACT OF THE STUDY: The lack of availability of water quality data constrains efforts to monitor, evaluate and improve the safety of water and sanitation infrastructure in underserved settings. Current water testing methods are not scalable because of laboratory and cost constraints. Our findings indicate the LCPA or similar low-cost microbial tests could be useful in rapid water safety estimation, including via crowdsourcing.
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Agua Potable/microbiología , Agua Potable/normas , Monitoreo del Ambiente/métodos , Técnicas Microbiológicas/métodos , Microbiología del Agua , Abastecimiento de Agua/normas , Recuento de Colonia Microbiana , Monitoreo del Ambiente/economía , Monitoreo del Ambiente/normas , Escherichia coli/crecimiento & desarrollo , Humanos , India , Estudios Longitudinales , Técnicas Microbiológicas/economía , Técnicas Microbiológicas/normasRESUMEN
This study assessed the prevalence of involuntary movements among older inpatients with severe schizophrenia, many of whom had experienced a lifetime of illness and its treatment, and examined their neuropsychological correlates. The subjects of this study were 128 inpatients with a DSM-IV diagnosis of schizophrenia. They were assessed using the Abnormal Involuntary Movement Scale, the Mini-Mental State Examination for general cognitive impairment and the Executive Interview for executive dyscontrol; additionally, their medical records were reviewed in detail for treatment histories. Prevalence of involuntary movements was examined and their clinical correlates determined in relation to topography of movement disorder using logistic regression. In schizophrenia, prevalence of involuntary movements was: age <65years, 63%; 65-75years, 80%; >75years, 93%. The primary correlate both of overall and of orofacial movements was poor executive function, whereas the primary correlate of limb-trunkal movements was poor general cognitive function. On approaching the limits of human longevity following a lifetime trajectory of illness and its treatment, essentially 'all' patients with schizophrenia appear inherently vulnerable to the emergence of involuntary movements in topographically specific association with cognitive deficits.
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Discinesia Inducida por Medicamentos/etiología , Lóbulo Frontal/fisiopatología , Trastornos Psicomotores/etiología , Esquizofrenia/complicaciones , Esquizofrenia/fisiopatología , Adulto , Anciano , Anciano de 80 o más Años , Antipsicóticos/administración & dosificación , Antipsicóticos/efectos adversos , Trastornos del Conocimiento/diagnóstico , Trastornos del Conocimiento/etiología , Trastornos del Conocimiento/fisiopatología , Discinesia Inducida por Medicamentos/diagnóstico , Femenino , Humanos , Masculino , Persona de Mediana Edad , Pruebas Neuropsicológicas , Trastornos Psicomotores/diagnóstico , Esquizofrenia/tratamiento farmacológico , Índice de Severidad de la EnfermedadRESUMEN
Timing of intervention with antipsychotic medication may influence long-term outcome in schizophrenia in a manner that is poorly understood. This study evaluated psychopathology, its factor structure, and cognitive dysfunction in older patients with chronic schizophrenia in relation to the intervals from onset of psychosis to initiation of treatment with antipsychotics, and from initiation of antipsychotic treatment to current assessments. The subjects were 129 patients with schizophrenia, many of whom became ill in the preneuroleptic era. Their current psychopathology was assessed using the Positive and Negative Syndrome Scale, and its factor structure examined using principal component analysis. Current general and executive cognitive function was evaluated using the Mini-Mental State Examination and the Executive Interview, respectively. Using multiple regression modelling, increasing duration of initially unmedicated psychosis, but not the much longer duration of subsequently treated illness, was the primary predictor of psychomotor poverty (negative symptoms) but not of reality distortion or disorganisation over the three domains of psychopathology resolved; duration of initially unmedicated psychosis marginally predicted the severity of general, but not of executive, cognitive dysfunction. Delayed intervention with antipsychotics appears associated with poorer long-term course in terms of increased severity of psychopathology in the psychomotor poverty domain.
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Antipsicóticos/uso terapéutico , Esquizofrenia/tratamiento farmacológico , Esquizofrenia/fisiopatología , Psicología del Esquizofrénico , Anciano , Enfermedad Crónica , Trastornos del Conocimiento/diagnóstico , Trastornos del Conocimiento/etiología , Femenino , Humanos , Masculino , Pruebas Neuropsicológicas , Trastornos Psicomotores/diagnóstico , Trastornos Psicomotores/etiología , Trastornos Psicóticos/diagnóstico , Trastornos Psicóticos/etiología , Índice de Severidad de la Enfermedad , Factores de TiempoRESUMEN
This phenomenological study describes the essential structure of the lived experience of adult male survivors of childhood sexual abuse by clergy (AMSCSABC). A purposive sample of seven AMSCSABC related their subjective experiences in semistructured interviews. Colaizzi's (1978) phenomenological method was used for data analysis. Survivors describe a bifurcated rage and spiritual distress that pervades their entire "lifebeing." Learning about AMSCSABC will assist nurses to identify potential risk factors of childhood sexual abuse by clergy (CSABC), design prevention strategies, and enhance empathy for a healing relationship.
Asunto(s)
Adaptación Psicológica , Abuso Sexual Infantil/psicología , Clero , Sobrevivientes/psicología , Adulto , Ira , Niño , Abuso Sexual Infantil/prevención & control , Clero/psicología , Mecanismos de Defensa , Empatía , Humanos , Masculino , Persona de Mediana Edad , Modelos Psicológicos , Relaciones Enfermero-Paciente , Investigación Metodológica en Enfermería , Factores de RiesgoRESUMEN
A novel series of 1,2-disubstituted indole, azaindole and benzimidazole derivatives possessing an amine moiety was identified as thrombin inhibitors. An indole with basic diamine moieties (12a) was the most potent thrombin inhibitor in the series with Kass= 197 x 10(6) L/mol.
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Anticoagulantes/síntesis química , Compuestos Aza/síntesis química , Bencimidazoles/síntesis química , Indoles/síntesis química , Trombina/antagonistas & inhibidores , Aminas/síntesis química , Aminas/química , Aminas/farmacología , Anticoagulantes/química , Anticoagulantes/farmacología , Compuestos Aza/química , Compuestos Aza/farmacología , Bencimidazoles/química , Bencimidazoles/farmacología , Diseño de Fármacos , Indoles/química , Indoles/farmacología , Cinética , Estructura Molecular , Relación Estructura-ActividadRESUMEN
Green fluorescent protein (GFP) was targeted into bacteriophage T4 heads and proheads as a probe of the internal environment. Targeting was accomplished with internal protein III (IPIII) fusion proteins or capsid targeting sequence (CTS)-tagged proteins, where CTS is the 10-amino acid residue CTS of IPIII. Recombinant phage T4[CTS/IPIII/GFP], T4[CTS/IPIII(T)GFP], and T4[CTS/GFP] packaged GFP fusion proteins and processed them at cleavage sites designated /. Steady-state and time-resolved fluorescence measurements suggest that packaged GFP is concentrated to a high density, that fusion protein IPIII(T)GFP occurs in a tightly clustered arrangement, and that the internal milieu of the phage head reduces rotational mobility of GFP. Phage, but not proheads, packaged with fusion protein IPIII(T)GFP gave an unexpectedly lower anisotropy than phage and proheads packaged with GFP, which suggests IPIII(T)GFP is bound to DNA in a manner that causes close associations between GFP molecules resulting in homotransfer between fluorophores within packaged phage. Targeting of reporter proteins into active virions is a promising approach for determining the structure of the condensed DNA, and properties of encapsidated viral enzymes.
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Bacteriófago T4/química , Bacteriófago T4/genética , Polarización de Fluorescencia , Proteínas Luminiscentes/genética , Bacteriófago T4/metabolismo , Cápside/metabolismo , Sondas de ADN , ADN Viral/química , ADN Viral/genética , ADN Viral/metabolismo , Proteínas Fluorescentes Verdes , Proteínas Luminiscentes/metabolismo , Conformación de Ácido Nucleico , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/metabolismo , Ensamble de VirusRESUMEN
6-[4-Amidinobenzoyl]amino]-tetralone-2-acetic acid is a potent antagonist of GPIIb-IIIa. Substitution in the meta position of the benzamidine, or replacement with a heteroaryl amidine was tolerated in this series. Use of an acyl-linked 4-alkyl piperidine as an arginine isostere also provided active compounds. Compounds from this series provided substantial systemic exposure in the rat following oral administration.
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Acetatos/metabolismo , Amidinas/metabolismo , Complejo GPIIb-IIIa de Glicoproteína Plaquetaria/antagonistas & inhibidores , Tetralonas , Adenosina Difosfato/farmacología , Animales , Arginina/química , Benzamidinas/química , Disponibilidad Biológica , Evaluación de Medicamentos , Ensayo de Inmunoadsorción Enzimática , Fibrinógeno/metabolismo , Humanos , Concentración 50 Inhibidora , Modelos Moleculares , Agregación Plaquetaria/efectos de los fármacos , Complejo GPIIb-IIIa de Glicoproteína Plaquetaria/metabolismo , Profármacos/síntesis química , Profármacos/química , Profármacos/farmacocinética , Unión Proteica/efectos de los fármacos , Estructura Secundaria de Proteína , Ratas , EstereoisomerismoRESUMEN
Each year, 19 million clients are treated for depressive episodes lasting 6 to 12 months. With depression's economic costs between 16 to 43 billion dollars annually, managed care has reduced clinician time, currently averaging 25 minutes per visit. The purpose of this phenomenological study is to describe the essential structure of the lived experience of depressed women who enter therapy and experience Watson's actual caring occasion (ACO) within the transpersonal caring relationship (TCR). A purposive sample of 11 depressed women related their subjective experiences in therapy over 6 months. Spiegelberg's phenomenological method was used for data analysis of 110 pages of therapist's notes transcribed verbatim. Five essential themes emerged from data analysis. All participants stated Watson's ACO caused them to persist in treatment and adopt health-seeking behaviors. This finding supports the expense of appropriate clinician time for holistic healing in the TCR.
Asunto(s)
Trastorno Depresivo/enfermería , Trastorno Depresivo/psicología , Empatía , Participación del Paciente , Adulto , Anécdotas como Asunto , Trastorno Depresivo/terapia , Femenino , Humanos , Persona de Mediana Edad , Satisfacción del Paciente , Relaciones Profesional-Paciente , Encuestas y Cuestionarios , Estados Unidos , Salud de la MujerRESUMEN
Disubstituted isoquinolones 2 and 3 have affinity for GPIIb-IIIa and represent leads for further structural evaluation. Structure-activity studies centered on the bicyclic beta-turn mimic contained in these molecules indicated that this moiety could accommodate a variety of modifications. Specifically, monocyclic, 6, 5-bicyclic, and 6,7-bicyclic structures provide compounds with affinity for GPIIb-IIIa. Within the 6,6-series, isoquinoline, tetralin, tetralone, and benzopyran nuclei yield potent antagonists that are specific for GPIIb-IIIa. Attachment of the arginine isostere (benzamidine) to the supporting nucleus can be accomplished with an ether or amide linkage, although the latter enhances activity. Several compounds in this series provided measurable blood levels after oral dosing. Conversion of the acid moiety in these molecules to an ester generally provided compounds which gave greater systemic exposure after oral administration. Absolute bioavailabilities in the rat for the ethyl ester prodrug derivatives of the tetralin, tetralone, and benzopyran analogues of 3 were 28%, 23%, and 24%, respectively.
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Benzopiranos/síntesis química , Isoquinolinas/síntesis química , Oligopéptidos/química , Complejo GPIIb-IIIa de Glicoproteína Plaquetaria/metabolismo , Tetrahidronaftalenos/síntesis química , Administración Oral , Animales , Benzopiranos/química , Benzopiranos/farmacocinética , Benzopiranos/farmacología , Unión Competitiva , Disponibilidad Biológica , Ensayo de Inmunoadsorción Enzimática , Cobayas , Humanos , Isoquinolinas/química , Isoquinolinas/farmacocinética , Isoquinolinas/farmacología , Imitación Molecular , Agregación Plaquetaria/efectos de los fármacos , Complejo GPIIb-IIIa de Glicoproteína Plaquetaria/antagonistas & inhibidores , Estructura Secundaria de Proteína , Ratas , Relación Estructura-Actividad , Tetrahidronaftalenos/química , Tetrahidronaftalenos/farmacocinética , Tetrahidronaftalenos/farmacologíaRESUMEN
A bacteriophage T4-derived protein expression, packaging and processing system was used to create recombinant phage that encode, produce and package a protein composed of human HIV-1 protease fused to green fluorescent protein (GFP). The fusion protein is targeted within the phage capsid by an N-terminal capsid targeting sequence (CTS), which is cleaved through proteolysis by the viral scaffold protease P21. The fusion protein is designated CTS [symbol see text] GFP:PR. The [symbol see text] symbol indicates the linkage peptide sequence leu(ile)-N-glu that is cleaved by the T4 head morphogenetic proteinase gp21 during head maturation. The fusion protein is fluorescent and has protease activity as detected by the appearance of the expected substrate cleavage product on a Western blot. CTS [symbol see text] GFP:PR packaging occurs at about 200 molecules per phage particle. The CTS [symbol see text] GFP:PR fusion protein, when protected within the phage capsid, has been maintained stably for over 16 months at 4 degrees C. Production and storage of fusion protein within the phage circumvents problems of toxicity and solubility encountered with E. coli expression systems. Because recombinant phage inhibit host proteolytic enzymes, foreign proteins are stabilized. This phage system packages and processes the fusion protein by means of the CTS. Proteins can be purified from the phage to give high yields of soluble, proteolytically processed protein. The T4 phage packaging system provides a novel means of identification, purification and long-term storage of toxic proteins whose folding and DNA-directed activities can be studied readily in vivo.
Asunto(s)
Bacteriófago T4/genética , Regulación Viral de la Expresión Génica , Proteasa del VIH/genética , Indicadores y Reactivos/metabolismo , Proteínas Luminiscentes/genética , Western Blotting , Escherichia coli/genética , Genes Reporteros , Proteínas Fluorescentes Verdes , Humanos , Microscopía Fluorescente , Plásmidos , Proteínas Recombinantes de Fusión/genética , beta-Galactosidasa/genéticaRESUMEN
The phage-derived expression, packaging, and processing (PEPP) system was used to target foreign proteins into the bacteriophage capsid to probe the intracapsid environment and the structure of packaged DNA. Small proteins with minimal requirements for activity were selected, staphylococcal nuclease (SN) and green fluorescent protein (GFP). These proteins were targeted into the T4 head by means of IPIII (internal protein III) fusions or CTS (capsid targeting sequence) fusions. Additional evidence is provided that foreign proteins are targeted into T4 by the N-terminal ten amino acid residue consensus CTS of IPIII identified in previous work. Fusion proteins were produced within host bacteria by expression from plasmids or by produc tion from recombinant phage carrying the fusion genes. Packaged fusion proteins CTS IPIII SN, CTS IPIII TSN, CTS IPIII GFP, CTS IPIII TGFP, and CTS GFP, where [symbol: see text] indicates a linkage peptide sequence Leu(Ile)-N-Glu cleaved by the T4 head morphogenetic proteinase gp21 during head maturation, are observed to exhibit intracapsid activity. SN activity within the head is demonstrated by loss of phage viability and by digested genomic DNA patterns visualized by gel electrophoresis when viable phage are incubated in Ca2+. Green fluorescent phage result immediately after packaging GFP produced at 30 degreesC and below, and continue to give green fluorescence under 470 nm light after CsCl purification. Non-fluorescent GFP-fusions are produced in bacteria at 37 degreesC, and phage packaged with these proteins achieve a fluorescent state after incubation for several months at 4 degreesC. GFP-packaged phage and proheads analyzed by fluorescence spectroscopy show that the mature head and the DNA-empty prohead package identical numbers of GFP-fusion proteins. Encapsidated GFP and SN can be injected into bacteria and rapidly exhibit intracellular activity. In vivo SN digestion of encapsidated DNA gives an intriguing pattern of DNA fragments by gel analysis, predominantly a repeat pattern of 160 bp multiples, reminiscent of a nucleosome digestion ladder, This quasi-limit DNA digestion pattern, reached >100-fold more slowly than the loss of titer, is invariant over a range
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Bacteriófago T4/metabolismo , ADN Viral/química , ADN Viral/metabolismo , Proteínas Recombinantes de Fusión/metabolismo , Bacteriófago T4/química , Bacteriófago T4/genética , Secuencia de Bases , Cápside/metabolismo , Cartilla de ADN/genética , ADN Viral/genética , Vectores Genéticos , Proteínas Fluorescentes Verdes , Cinética , Proteínas Luminiscentes/genética , Proteínas Luminiscentes/metabolismo , Nucleasa Microcócica/genética , Nucleasa Microcócica/metabolismo , Modelos Moleculares , Conformación de Ácido Nucleico , Reacción en Cadena de la Polimerasa , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/genéticaRESUMEN
Primary adrenal lymphoma is a rare entity, with only 16 cases reported in the last 40 yr. Although 67Ga scintigraphy has been extensively used to evaluate patients with other types of lymphomas, there are no reports of its use in patients with this disease entity. A man with primary adrenal lymphoma and no evidence of extraadrenal spread who was evaluated from presentation to remission with gallium scintigraphy and CT is presented. Gallium scintigraphy was valuable in assessing response to therapy.
Asunto(s)
Neoplasias de las Glándulas Suprarrenales/diagnóstico por imagen , Radioisótopos de Galio , Linfoma de Células B/diagnóstico por imagen , Adulto , Citratos , Galio , Humanos , Masculino , Cintigrafía , Tomografía Computarizada por Rayos XRESUMEN
Cyclic heptapeptide 1, which contains an Arg-Gly-Asp sequence, has good affinity for the platelet receptor GPIIb-IIIa and was chosen for study by 1H NMR techniques. The key RGD sequence of this molecule was found to reside in a conformationally defined type II' Gly-Asp beta-turn, and this information was used in the design of simple non-peptide RGD mimics. Disubstituted isoquinolones, bearing an acidic side chain at position 2 and a basic side chain at position 6, were prepared and were found to have modest affinity for GPIIb-IIIa. Systematic modification of the basic residue contained in these molecules yielded compounds with high affinity for GPIIb-IIIa.
Asunto(s)
Oligopéptidos/química , Complejo GPIIb-IIIa de Glicoproteína Plaquetaria/antagonistas & inhibidores , Ácido Aspártico , Diseño de Fármacos , Ensayo de Inmunoadsorción Enzimática , Glicina , Humanos , Modelos Químicos , Modelos Moleculares , Imitación Molecular , Oligopéptidos/farmacología , Agregación Plaquetaria , Estructura Secundaria de Proteína , Relación Estructura-ActividadRESUMEN
Information poverty is one of the most significant obstacles to better health in the developing world. To address this problem, SatelLife, a Boston-based nonprofit organization has created HealthNet, a computer-based network linking healthworkers throughout the world to facilitate the exchange of health-related information. Using low-cost, sustainable technologies, SatelLife provides both electronic mail communications services and a variety of information services including electronic publications, electronic access to remote databases, and ongoing electronic conferences on topics of special relevance to health in the developing world. The article cautions that simply bringing high-speed Internet access to the developing world will not solve problems of information poverty, because such technology will be prohibitively expensive for all but the elite private sector and because of the need to provide targeted, high-quality information, not simply access to the vast, disorganized universe of information available on the Internet.
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Redes de Comunicación de Computadores/organización & administración , Países en Desarrollo , Personal de Salud , Boston , Accesibilidad a los Servicios de Salud , Necesidades y Demandas de Servicios de Salud , Gestión de la Información , Informática Médica , Organizaciones sin Fines de Lucro , PobrezaRESUMEN
PURPOSE: Cytomorphometric analysis of uveal melanoma has focused on nucleolar characteristics, although nuclear measurements are easier and more reproducible. In this study, we examine a restricted set of nuclear and nucleolar cytomorphometric variables in the search for the most quantifiable and reproducible prognostic parameter among these and discover a means of assessing the malignancy of tumors within the different Callender subgroups. METHODS: Uveal melanomas from 94 patients with a minimum follow-up of 5 years underwent nuclear and nucleolar morphometric analysis using a digitized interactive video overlay system. Statistical analysis included univariate and multivariate survival analysis on morphometric variables and classic prognostic parameters (such as Callender cell type, size, etc.). RESULTS: Univariate analysis of cytomorphometric parameters showed the standard deviation of nuclear area (NASD) to be the most significant variable (P < 0.0001: Mantel-Cox, 15.2). This was followed by the difference between nuclear and nucleolar areas (P < 0.002; Mantel-Cox, 9.5), the standard deviation of nucleolar area (SDNA) (P < 0.01; Mantel-Cox, 6.4), and the standard deviation of the shortest nuclear axis (P < 0.02; Mantel-Cox, 6.4). There was a trend for significance with the mean of the ten largest nucleoli (P < 0.07). Multivariate analysis of clinical and cytomorphometric variables showed improvement of prognostic prediction with a combination of the NASD, largest tumor dimension, and glaucoma (P < 0.0001; Mantel-Cox, 26.3). Callender cell type was strongly correlated with the NASD (P < 0.0002), the mean axis ratio of the nuclear area (P < 0.0007), and the mean of the ten largest nucleoli (P < 0.0007). CONCLUSION: Cytomorphometric analysis of uveal melanoma should include nuclear area and may identify tumors of differing malignancy within the same Callender class.
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Melanoma/clasificación , Melanoma/patología , Neoplasias de la Úvea/clasificación , Neoplasias de la Úvea/patología , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Enucleación del Ojo , Humanos , Procesamiento de Imagen Asistido por Computador , Melanoma/mortalidad , Persona de Mediana Edad , Pronóstico , Reproducibilidad de los Resultados , Tasa de Supervivencia , Neoplasias de la Úvea/mortalidadRESUMEN
A membrane-independent morphogenetic viral signal peptide is identified within bacteriophage T4 internal protein III (IPIII). Utilizing a phagederived expression-packaging-processing system, which packages foreign proteins fused with IPIII into the phage capsid, a synthetic cleavage site introduced at the C terminus of IPIII, is demonstrated to be functional and permits processing of fusion proteins. IPIII, which possesses a native P21 cleavage site at its N terminus, is altered to possess a second P21 cleavage site at its C terminus where cleavage occurs by means of the scaffold proteinase P21 within the capsid. The altered IPIII was inserted into an expression vector to permit the creation of fusion proteins with staphylococcal nuclease, EcoRI endonuclease, beta-globin, and luciferase. Western immunoblot analysis of packaged T4eG326 indicates that the IPIII:fusion-proteins are packaged into phage and proteolytically processed, thus the synthetic P21 cleavage site positioned at the C terminus of IPIII is demonstrated to be functional, and 20 to 200 protein molecules are packaged per capsid. Truncation experiments identified the minimal portion of IPIII required to achieve targeting into the phage capsid as a ten amino acid residue from the N terminus, which includes the N-terminal methionine residue and the proteinase P21 cleavage site, designated the CTS (capsid targeting sequence). The addition of the CTS to a fragment of luciferase permits the protein to be packaged and processed, which demonstrates that the CTS is by itself sufficient to target foreign protein to the capsid. The imputed dual function of the CTS is supported by site-directed PCR mutagenesis, which reveals two functionally separate domains of the CTS for targeting and processing. The CTS appears to function in a core-related targeting mechanism that directs a polymorphic set of proteins into the T-even capsid or scaffold. Although structure formation is often assumed to involve extended protein interfaces, the analysis shows that a limited but specific sequence, the CTS, drives the interaction required to achieve targeting.
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Bacteriófago T4/genética , Proteínas de la Cápside , Cápside/genética , Vectores Genéticos , Proteínas Recombinantes de Fusión/genética , Secuencia de Aminoácidos , Bacteriófago T4/metabolismo , Secuencia de Bases , Cápside/metabolismo , Datos de Secuencia Molecular , Mutagénesis , Análisis de SecuenciaRESUMEN
The most common method of exocrine drainage in pancreatic transplantation is urinary drainage. With the shift from enteric drainage and duct occlusion techniques, there has been a concomitant shift from intraabdominal to urological complications. We present a case in which a simultaneous pancreas-kidney (SPK) transplant recipient developed urethral disruption and associated bladder outlet obstruction 11 months following surgery.
Asunto(s)
Quimotripsina/orina , Diabetes Mellitus Tipo 1/cirugía , Nefropatías Diabéticas/cirugía , Trasplante de Riñón/fisiología , Trasplante de Páncreas/fisiología , Complicaciones Posoperatorias/diagnóstico por imagen , Tripsina/orina , Enfermedades Uretrales/diagnóstico por imagen , Obstrucción del Cuello de la Vejiga Urinaria/diagnóstico por imagen , Fístula Urinaria/diagnóstico por imagen , Urografía , Adulto , Cistostomía , Diabetes Mellitus Tipo 1/diagnóstico por imagen , Diabetes Mellitus Tipo 1/enzimología , Nefropatías Diabéticas/diagnóstico por imagen , Nefropatías Diabéticas/enzimología , Extravasación de Materiales Terapéuticos y Diagnósticos/diagnóstico por imagen , Extravasación de Materiales Terapéuticos y Diagnósticos/enzimología , Estudios de Seguimiento , Humanos , Masculino , Complicaciones Posoperatorias/enzimología , Enfermedades Uretrales/enzimología , Obstrucción del Cuello de la Vejiga Urinaria/enzimología , Fístula Urinaria/enzimologíaRESUMEN
DNA coding for bacteriophage T7 RNA polymerase (T7-RNAP) was inserted into a positive selection-vector form of the T4 genome, placing it under the control of bacteriophage T4 ipIII promoters. The recombinant T4::T7-RNAP fusion phage retained infectivity and produced T7-RNAP in infected cells. Fusion genes were constructed by insertion into a plasmid containing an iPIII (encoding internal protein III) target portion and a bacteriophage T7 promoter region. When Escherichia coli cells containing the plasmid were infected with the T4::T7-RNAP re-phage, the bacteria produced fusion protein at high levels. The newly synthesized T4::T7-RNAP re-phage progeny package and process the fusion protein into the phage capsid during head morphogenesis. In this paper, we demonstrate that truncated T4 internal protein IPIII, human IPIII::beta Glo (beta-globin) fusion protein, E. coli IPIII::beta Glo::beta Gal (beta-galactosidase) triple-fusion protein and IPIII::V3 fusion protein (human immunodeficiency virus envelope protein gp120 V3 region) are expressed at high levels by T4::T7-RNAP induction. With IPIII::beta Glo, expression-packaging-processing (EPP) occurs simultaneously with T4::T7-RNAP re-phage infection. We also demonstrate that T4::T7-RNAP re-phage stabilize unstable proteins such as the X90 fragment of beta Gal, thought to be degraded by the lon protease. An unstable 20-kDa fragment of the large subunit of human cytochrome b558, an integral membrane protein in phagocytes, is subject to proteolytic degradation even when produced in the lon-deficient BL21 strain. However, upon induction with T4::T7-RNAP re-phage, the 20-kDa protein is produced intact.(ABSTRACT TRUNCATED AT 250 WORDS)
Asunto(s)
Bacteriófago T4/genética , Proteínas de la Cápside , Vectores Genéticos , NADPH Oxidasas , Procesamiento Proteico-Postraduccional , Proteínas Recombinantes de Fusión/biosíntesis , Bacteriófago T4/crecimiento & desarrollo , Bacteriófago T7/enzimología , Cápside/biosíntesis , Cápside/genética , Grupo Citocromo b/biosíntesis , Grupo Citocromo b/genética , ARN Polimerasas Dirigidas por ADN/genética , Endopeptidasas/metabolismo , Vectores Genéticos/efectos de los fármacos , Globinas/biosíntesis , Globinas/genética , Proteína gp120 de Envoltorio del VIH/biosíntesis , Proteína gp120 de Envoltorio del VIH/genética , Humanos , Proteínas ViralesRESUMEN
Deoxyribonucleic acid (DNA) ploidy was quantified in 84 ocular melanomas (median follow-up, 11-years) by flow cytometry, CAS 200 interactive image analysis, and Pathology Image Processing Environment (PIPE; Department of Quantitative Pathology, Free University, Amsterdam, The Netherlands) automatic image analysis (75). Overall, 32.1% of the melanomas were aneuploid, 2.3% were tetraploid, and 66.6% were diploid. Pathology Image Processing Environment analysis estimated DNA ploidy in 12 tumors that were unprocessable by flow cytometry. Of 10 tumors that were diploid by flow cytometry, PIPE detected stemline aneuploidy in five and some aneuploid cells in five more. Seven tumors were aneuploid by PIPE but diploid by CAS 200, five of which contained occasional DNA aneuploid cells on the CAS 200 histograms. Pathology Image Processing Environment analysis quantified tumor samples with an average of 500 (250 to 1,050) cells in less than 10 minutes. All cells classified as spindle A according to the Callender system (more than 10,000) were diploid. Spindle B and epithelioid cells occupied both diploid and aneuploid peaks on the DNA histograms. Dioxyribonucleic acid variables did not correlate with established prognosticators, such as Callender cell type, largest tumor dimension, or glaucoma, nor did they reach significance by univariate and multivariate analyses. The value of these findings as a diagnostic support in uveal melanoma, particularly in combination with fine needle aspiration biopsy, is discussed.
Asunto(s)
ADN de Neoplasias/análisis , ADN de Neoplasias/genética , Neoplasias del Ojo/genética , Melanoma/genética , Ploidias , Neoplasias del Ojo/patología , Citometría de Flujo , Humanos , Procesamiento de Imagen Asistido por Computador , Melanoma/patología , Análisis de SupervivenciaRESUMEN
Syntactic structure analysis was carried out successfully on 92 paraffin embedded uveal melanomas, taken from patients with a minimum follow up of 5 years. This simple, fast, and reproducible method of describing the tumour architecture has been significantly correlated with malignancy in tumours from several sites. Paraffin sections 5 microns thick, were cut and stained with haematoxylin and eosin. Tumours were classified according to a modification of the Callender classification. A minimum spanning tree (MST), using the centre points of tumour nuclei, was constructed in five randomly chosen fields with an interactive digitising video overlay system. Ten syntactic structure features were derived from each MST; subsequently, the mean and standard deviation of the five fields analysed were calculated for further statistical analysis. Reproducibility was acceptable with a mean correlation coefficient of 0.70. In univariate survival analysis, the percentage of points with three neighbours yielded prognostic significance (p < 0.05). Minimum spanning tree variables were compared (chi 2 test) with classic tumour prognosticators and there was a significant correlation between Callender cell type and the following MST parameters: mean number of points (p < 0.003); MST length (p < 0.003); mean line length (p < 0.01); number of nuclei with one neighbour (p < 0.004); number of nuclei with two neighbours (p < 0.02), and number of nuclei with three neighbours (p < 0.005). Syntactic structure analysis is an evolving technique, but may be able to mathematically (and reproducibly) describe melanoma architecture across the spectrum of the Callender classes. This would also allow architectural grading of tumours within the specific Callender groups, providing more precise prognostic information. Further modifications of this technique are necessary to optimise prognostic potential when applied to uveal melanomas.