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1.
Cardiovasc Intervent Radiol ; 42(12): 1687-1694, 2019 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-31531691

RESUMEN

PURPOSE: To evaluate the incidence of acute renal failure and chronic kidney disease due to occlusion of accessory renal arteries during endovascular aneurysm repair of infrarenal abdominal aortic aneurysm. MATERIAL AND METHODS: We retrospectively reviewed the course of 181 patients (mean age, 71, SD ± 9  years) who underwent EVAR of infrarenal abdominal aortic aneurysm. The renal vessel anatomy was analyzed in all pre- and postoperative CT scans. Diameter and origin of accessory renal arteries were evaluated. Renal function was determined by pre- and postoperative serum creatinine and eGFR levels. Long-term follow-up (>3 months) of patients was available in 121 cases (66.9%). Acute kidney injury and chronic kidney failure were defined according to guidelines of "Kidney Disease: Improving Global Outcomes" (KDIGO). RESULTS: In 65 of 181 patients (33.9%), 82 accessory renal arteries were identified preoperatively. In 19 of 181 patients (10.5%), one or more accessory renal arteries were covered and subsequently occluded by the implanted stent-graft device. Neither acute kidney injury (10.3% vs 12.5%; p = .785) nor chronic kidney disease (10.7% vs 15.38%; p = .452) was detected significantly more often in patients with covered accessory renal artery. The only significant predictor of acute kidney injury was the preoperative serum creatinine level (1.12 mg/dl vs. 0.98 mg/dl; p = .03). Significant predictors for chronic kidney disease were preoperative serum creatinine, eGFR, and impaired renal function (p < .001). CONCLUSION: Coverage of accessory renal artery due to stent-graft does not lead either to temporary acute kidney injury after endovascular aneurysm repair or to chronic kidney disease. LEVEL OF EVIDENCE: Level II b.


Asunto(s)
Lesión Renal Aguda/epidemiología , Aneurisma de la Aorta Abdominal/cirugía , Implantación de Prótesis Vascular/métodos , Complicaciones Posoperatorias/epidemiología , Arteria Renal/cirugía , Insuficiencia Renal Crónica/epidemiología , Anciano , Aneurisma de la Aorta Abdominal/epidemiología , Procedimientos Endovasculares/métodos , Femenino , Estudios de Seguimiento , Humanos , Masculino , Estudios Retrospectivos , Resultado del Tratamiento
2.
Chirurg ; 87(9): 744-750, 2016 Sep.
Artículo en Alemán | MEDLINE | ID: mdl-27495164

RESUMEN

The 1­year incisional hernia rate of 9-30 % has been tolerated for decades. Even in the 1970s and 1980s there was evidence that supported reducing suture tension. Recently, the traditional 4:1 relationship between suture and wound length, which has been passed on for years, has been questioned. After first experimental and clinical data suggested an advantage by reducing the width and interval of stitches by 50 %, the prospective randomized STITCH study has now provided evidence by significantly lowering the 1­year hernia rate from 21 % to 13 %. For surgeons this means less of a revolution and more of an innovative evolution of a long-established technique. Before introduction of the technique quality assurance must be carried out with documentation of performance indicators (e.g. number of stitches, length of thread incorporated and wound length).


Asunto(s)
Técnicas de Cierre de Herida Abdominal/educación , Educación Médica Continua , Técnicas de Cierre de Herida Abdominal/normas , Humanos , Hernia Incisional/prevención & control , Estudios Prospectivos , Garantía de la Calidad de Atención de Salud/normas , Ensayos Clínicos Controlados Aleatorios como Asunto , Tejido Subcutáneo/cirugía , Técnicas de Sutura/educación
4.
Br J Surg ; 102(7): 805-12, 2015 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-25877255

RESUMEN

BACKGROUND: Epidural analgesia (EDA) is a common analgesia regimen in liver resection, and is accompanied by sympathicolysis, peripheral vasodilatation and hypotension in the context of deliberate intraoperative low central venous pressure. This associated fall in mean arterial pressure may compromise renal blood pressure autoregulation and lead to acute kidney injury (AKI). This study investigated whether EDA is a risk factor for postoperative AKI after liver surgery. METHODS: The incidence of AKI was investigated retrospectively in patients who underwent liver resection with or without EDA between 2002 and 2012. Univariable and multivariable analyses were performed including recognized preoperative and intraoperative predictors of posthepatectomy renal failure. RESULTS: A series of 1153 patients was investigated. AKI occurred in 8·2 per cent of patients and was associated with increased morbidity (71 versus 47·3 per cent; P = 0·003) and mortality (21 versus 0·3 per cent; P < 0·001) rates. The incidence of AKI was significantly higher in the EDA group (10·1 versus 3·7 per cent; P = 0·003). Although there was no significant difference in the incidence of AKI between patients undergoing minor hepatectomy with or without EDA (5·2 versus 2·7 per cent; P = 0·421), a substantial difference in AKI rates occurred in patients undergoing major hepatectomy (13·8 versus 5·0 per cent; P = 0·025). In multivariable analysis, EDA remained an independent risk factor for AKI after hepatectomy (P = 0·040). CONCLUSION: EDA may be a risk factor for postoperative AKI after major hepatectomy.


Asunto(s)
Lesión Renal Aguda/epidemiología , Analgesia Epidural/efectos adversos , Tasa de Filtración Glomerular/fisiología , Hepatectomía/efectos adversos , Complicaciones Posoperatorias/epidemiología , Lesión Renal Aguda/etiología , Lesión Renal Aguda/fisiopatología , Estudios de Seguimiento , Incidencia , Pruebas de Función Renal , Neoplasias Hepáticas/cirugía , Periodo Perioperatorio , Complicaciones Posoperatorias/etiología , Complicaciones Posoperatorias/fisiopatología , Pronóstico , Estudios Retrospectivos , Factores de Riesgo , Suiza/epidemiología
5.
Soc Psychiatry Psychiatr Epidemiol ; 48(7): 1021-31, 2013 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-23644725

RESUMEN

PURPOSE: The ABC Schizophrenia study, led by a single research team, investigated a schizophrenia sample systematically over quarter of a century. This paper summarises results from 1996 onwards. The initial goals were to explain the considerably higher age at first admission in women, and to obtain precise information on the onset and early course of schizophrenia as a prerequisite for early intervention. METHOD: The study was hypothesis-driven. People with schizophrenia were compared in the prodrome and at first admission to those with unipolar depression and to healthy controls. We analysed the medium-term (5-year) and the long-term (12-year) course of schizophrenia, its symptom dimensions, social parameters and predictors. SAMPLES: (1) 276 population-based first admissions (232 first episodes) of schizophrenia (age range 12-59 years); (2) a subsample of 130 first admissions for schizophrenia; (3) 130 first admissions for unipolar depression; (4) 130 healthy population controls and (5) 1,109 consecutive first admissions for schizophrenia spectrum disorder without an age limit. RESULTS: The prodromal stages of schizophrenia and depression were very similar until positive symptoms appeared. The most frequent symptom in schizophrenia was depressed mood. The course of psychosis from prodrome to 12 years following first admission was very variable. From 5 to 12 years after first admission the course was characterised by irregular exacerbations of the main symptom dimensions, with no overall deterioration or improvement. CONCLUSIONS: Schizophrenic psychosis and severe affective disorder, rather than representing discrete illnesses, probably mark different stages in the manifestation of psychopathology produced by various degrees of brain dysfunction.


Asunto(s)
Esquizofrenia/diagnóstico , Adaptación Psicológica , Adolescente , Adulto , Factores de Edad , Niño , Comorbilidad , Trastorno Depresivo/diagnóstico , Progresión de la Enfermedad , Femenino , Hospitalización , Humanos , Masculino , Persona de Mediana Edad , Síntomas Prodrómicos , Esquizofrenia/epidemiología , Psicología del Esquizofrénico , Trastornos Relacionados con Sustancias/epidemiología , Factores de Tiempo , Adulto Joven
6.
Nervenarzt ; 84(9): 1093-4, 1096-103, 2013 Sep.
Artículo en Alemán | MEDLINE | ID: mdl-23695002

RESUMEN

BACKGROUND: The ABC schizophrenia study conducted by the same team over 25 years initially aimed at illuminating the onset, prodromal stage and sex differences in age at first hospitalization in schizophrenia. New hypotheses were systematically generated from the results achieved. METHODS: A population-based sample of 276 first admission cases (232 first episodes, age 12-59 years), including a subsample of 130 first admissions (115 first episodes), were assessed to study prodromal stage, first illness episode, medium and long-term course and symptom dimensions in schizophrenia. The samples were compared with age and sex-matched healthy controls and with patients first admitted for unipolar depression. A total of 1,109 consecutive first admissions for schizophrenia spectrum disorders independent from the other study samples were assessed to study changes in symptomatology across the age range. RESULTS: Before the onset of psychotic symptoms the prodromal stages of schizophrenia and severe and moderately severe depression are difficult to distinguish. The most frequent symptom in the course of schizophrenia, depressed mood, also represents the most frequent initial symptom in both disorders. Prodromal depression is a predictor of more depressive and positive symptoms in the first episode but not in the further course of the illness. Psychosis incidence for men, diagnosed according to ICD 9 (295, 297, 298.3/4), shows a pronounced peak at age 15-24 years, for women a lower peak at age 15-29 years and a second, still lower peak at the menopausal age of 45-49 years. The explanation, confirmed in animal experiments, lies in a protective effect of estrogen due to reduced D2 receptor sensitivity. The effect is antagonized by an elevated genetic risk. Functional and social impairment emerge even at the prodromal stage and the severity depends on sex and social status. Young men with schizophrenia show a less favorable social course because of the earlier age of onset and socially adverse illness behavior. Late onset is associated with a milder, primarily paranoid symptomatology and less severe social impairment. Schizophrenia is a disorder of all ages showing roughly equal life time incidence rates for men and women but considerable difference in certain periods of age. The symptom dimensions show a plateau-like course 2-5 years after the first episode. Hidden behind this picture are irregular symptom exacerbations which vary in duration. Schizophrenia conveys the picture of recurrent vulnerability to crisis and not of a stable residual state of disordered brain development or of a progressive neurodegenerative process.


Asunto(s)
Depresión/diagnóstico , Depresión/epidemiología , Trastornos Psicóticos/diagnóstico , Trastornos Psicóticos/epidemiología , Esquizofrenia/diagnóstico , Esquizofrenia/epidemiología , Adolescente , Adulto , Distribución por Edad , Causalidad , Niño , Comorbilidad , Diagnóstico Diferencial , Femenino , Alemania/epidemiología , Humanos , Masculino , Persona de Mediana Edad , Prevalencia , Factores de Riesgo , Distribución por Sexo , Síndrome , Adulto Joven
7.
Lupus ; 22(3): 297-306, 2013 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-23439470

RESUMEN

Integrin very late antigen-4 (VLA4) is induced during inflammation and can regulate monocyte migration. It has been implicated in atherogenesis, a significant concern in systemic lupus erythematosus (SLE). The aim of this study was to define VLA4 expression in SLE monocytes. Flow cytometry, reverse transcription polymerase chain reaction, Western blotting, and immunohistochemistry staining with confocal microscopy were used to evaluate VLA4 expression in SLE patients and controls. We found elevated expression of VLA4 in SLE patients with significantly increased VLA4 staining intracellularly compared to control. Exposure of control monocytes to SLE sera or immune complexes led to increased intracellular expression, and immune complexes were capable of driving redistribution of surface VLA4 to the cytoplasm. Therefore, VLA4 was found to be subject to complex regulation with SLE sera driving both RNA expression and redistribution of protein. Stimulation of SLE monocytes with a VLA4 ligand induced significant TNFα expression, confirming a functional effect. This behavior may contribute to increased atherosclerosis and monocyte infiltrates in end organs.


Asunto(s)
Integrina alfa4beta1/inmunología , Lupus Eritematoso Sistémico/inmunología , Femenino , Humanos , Integrina alfa4beta1/biosíntesis , Monocitos/inmunología
8.
J Biotechnol ; 157(1): 140-7, 2012 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-21983234

RESUMEN

A large strain collection comprising antagonistic bacteria was screened for novel detergent proteases. Several strains displayed protease activity on agar plates containing skim milk but were inactive in liquid media. Encapsulation of cells in alginate beads induced protease production. Stenotrophomonas maltophilia emerged as best performer under washing conditions. For identification of wash-active proteases, four extracellular serine proteases called StmPr1, StmPr2, StmPr3 and StmPr4 were cloned. StmPr2 and StmPr4 were sufficiently overexpressed in E. coli. Expression of StmPr1 and StmPr3 resulted in unprocessed, insoluble protein. Truncation of most of the C-terminal domain which has been identified by enzyme modeling succeeded in expression of soluble, active StmPr1 but failed in case of StmPr3. From laundry application tests StmPr2 turned out to be a highly wash-active protease at 45°C. Specific activity of StmPr2 determined with suc-L-Ala-L-Ala-L-Pro-l-Phe-p-nitroanilide as the substrate was 17±2U/mg. In addition we determined the kinetic parameters and cleavage preferences of protease StmPr2.


Asunto(s)
Proteínas Bacterianas/biosíntesis , Escherichia coli/metabolismo , Proteínas Recombinantes/biosíntesis , Serina Proteasas/biosíntesis , Stenotrophomonas maltophilia/enzimología , Alginatos/química , Animales , Proteínas Bacterianas/química , Proteínas Bacterianas/genética , Medios de Cultivo/metabolismo , Detergentes/química , Escherichia coli/genética , Espacio Extracelular/metabolismo , Ácido Glucurónico/química , Ácidos Hexurónicos/química , Insulina/metabolismo , Leche/metabolismo , Datos de Secuencia Molecular , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Serina Proteasas/química , Serina Proteasas/genética , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Stenotrophomonas maltophilia/genética
9.
Br J Psychiatry ; 200(1): 22-9, 2012 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-22075649

RESUMEN

BACKGROUND: Young people with self-experienced cognitive thought and perception deficits (basic symptoms) may present with an early initial prodromal state (EIPS) of psychosis in which most of the disability and neurobiological deficits of schizophrenia have not yet occurred. AIMS: To investigate the effects of an integrated psychological intervention (IPI), combining individual cognitive-behavioural therapy, group skills training, cognitive remediation and multifamily psychoeducation, on the prevention of psychosis in the EIPS. METHOD: A randomised controlled, multicentre, parallel group trial of 12 months of IPI v. supportive counselling (trial registration number: NCT00204087). Primary outcome was progression to psychosis at 12- and 24-month follow-up. RESULTS: A total of 128 help-seeking out-patients in an EIPS were randomised. Integrated psychological intervention was superior to supportive counselling in preventing progression to psychosis at 12-month follow-up (3.2% v. 16.9%; P = 0.008) and at 24-month follow-up (6.3% v. 20.0%; P = 0.019). CONCLUSIONS: Integrated psychological intervention appears effective in delaying the onset of psychosis over a 24-month time period in people in an EIPS.


Asunto(s)
Terapia Cognitivo-Conductual/métodos , Progresión de la Enfermedad , Educación del Paciente como Asunto , Trastornos Psicóticos/prevención & control , Esquizofrenia/prevención & control , Psicología del Esquizofrénico , Adolescente , Adulto , Atención Ambulatoria , Consejo , Susceptibilidad a Enfermedades/psicología , Salud de la Familia , Femenino , Humanos , Estimación de Kaplan-Meier , Masculino , Estudios Prospectivos , Escalas de Valoración Psiquiátrica , Trastornos Psicóticos/patología , Trastornos Psicóticos/psicología , Factores de Tiempo , Resultado del Tratamiento , Adulto Joven
10.
Ultraschall Med ; 33(7): E339-E343, 2012 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-21882142

RESUMEN

PURPOSE: To assess the suitability of spinal ultrasound for the detection of spinal subdural hematoma in infants with sustained non-accidental trauma. MATERIALS AND METHODS: Six infants (mean age ± SD 3.3 ± 1.5 months) admitted to our hospital because of suspected non-accidental trauma were examined radiologically with ultrasound, CT and/or MRI and skeletal radiography. Twelve healthy infants (mean age ± SD 2.5 ± 1.4 months) in whom an ultrasound of the spine was performed to exclude spinal dysraphism served as controls. RESULTS: All six patients with non-accidental trauma (NAT) presented with cranial subdural hematoma visualized by ultrasound and CT scan or MRI. Spinal ultrasound detected echogenic effusions with floating particles that displaced the undulating arachnoidea from the dura mater spinalis in all six patients with NAT. The size of the spinal subdural hematoma varied and extended from the cervical spine to the cauda equina. The anatomic landmarks (dura mater spinalis, arachnoidea spinalis) were identified and confirmed the subdural location. All spinal subdural hematomas were asymptomatic and detected by diagnostic ultrasound. None of the infants had a pre-existing neurological or hemorrhagic disorder. The plain X-rays of the spine in these infants showed no osseous lesion. Spinal subdural hematoma was not observed in any of the controls. CONCLUSION: The presence of spinal subdural hematoma is a valuable sign of sustained non-accidental trauma in infants that can be quickly and easily detected using spinal ultrasound without the need for sedation or general anesthesia. Thus, spinal ultrasound should be part of the imaging examinations performed in infants with suspected abuse.


Asunto(s)
Síndrome del Niño Maltratado/diagnóstico por imagen , Hematoma Subdural Espinal/diagnóstico por imagen , Interpretación de Imagen Asistida por Computador , Daño Encefálico Crónico/diagnóstico por imagen , Ecoencefalografía , Femenino , Hematoma Intracraneal Subdural/diagnóstico por imagen , Humanos , Lactante , Imagen por Resonancia Magnética , Masculino , Sensibilidad y Especificidad , Hemorragia Subaracnoidea/diagnóstico por imagen , Tomografía Computarizada por Rayos X
11.
Microb Biotechnol ; 4(6): 767-76, 2011 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-21895993

RESUMEN

In the wide field of laundry and cleaning applications, there is an unbroken need for novel detergent proteases excelling in high stability and activity and a suitable substrate range. We demonstrated the large amount of highly diverse subtilase sequences present in metagenomic DNA by recovering 57 non-redundant subtilase sequence tags with degenerate primers. Furthermore, an activity- as well as a sequence homology-based screening of metagenomic DNA libraries was carried out, using alkaline soil and habitat enrichments as a source of DNA. In this way, 18 diverse full-length protease genes were recovered, sharing only 37-85% of their amino acid residues with already known protease genes. Active clones were biochemically characterized and subjected to a laundry application assay, leading to the identification of three promising detergent proteases. According to sequence similarity, two proteases (HP53 and HP70) can be classified as subtilases, while the third enzyme (HP23) belongs to chymotrypsin-like S1 serine proteases, a class of enzymes that has not yet been described for the use in laundry and cleaning applications.


Asunto(s)
Biotecnología/métodos , Metagenómica/métodos , Péptido Hidrolasas/genética , Péptido Hidrolasas/metabolismo , Secuencia de Aminoácidos , Análisis por Conglomerados , Detergentes/metabolismo , Inhibidores Enzimáticos/metabolismo , Estabilidad de Enzimas , Concentración de Iones de Hidrógeno , Datos de Secuencia Molecular , Peso Molecular , Péptido Hidrolasas/química , Péptido Hidrolasas/aislamiento & purificación , Filogenia , Análisis de Secuencia de ADN , Homología de Secuencia de Aminoácido , Microbiología del Suelo , Temperatura
12.
Mucosal Immunol ; 4(6): 658-70, 2011 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-21796113

RESUMEN

Oral infection of C57BL/6 mice with Toxoplasma gondii triggers severe necrosis in the ileum within 7-10 days of infection. Lesion development is mediated by Th-1 cytokines, CD4+ T cells, and subepithelial bacterial translocation. As such, these features share similarity to Crohn's disease. Recently, we uncovered a role for intraepithelial lymphocytes (IELs) in mediating pathology after Toxoplasma infection. We show here that αß and not γδ T-cell IELs mediate intestinal damage. By adoptive transfer of mucosal T cells into naive Rag1⁻/⁻ mice, we demonstrate that IELs do not function alone to cause inflammatory lesions, but act with CD4+ T lymphocytes from the lamina propria (LP). Furthermore, recipient mice pretreated with broad-spectrum antibiotics to eliminate intestinal flora resisted intestinal disease after transfer of IELs and LP lymphocytes. Our data provide valuable new insights into the mechanisms of intestinal inflammation, findings that have important implications for understanding human inflammatory bowel disease.


Asunto(s)
Linfocitos T CD4-Positivos/metabolismo , Enfermedad de Crohn/inmunología , Subgrupos de Linfocitos T/metabolismo , Toxoplasma/inmunología , Toxoplasmosis/inmunología , Traslado Adoptivo , Animales , Profilaxis Antibiótica , Linfocitos T CD4-Positivos/efectos de los fármacos , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/patología , Comunicación Celular , Enfermedad de Crohn/fisiopatología , Enfermedad de Crohn/prevención & control , Modelos Animales de Enfermedad , Humanos , Íleon/efectos de los fármacos , Íleon/inmunología , Íleon/microbiología , Inflamación , Mucosa Intestinal/patología , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Receptores de Antígenos de Linfocitos T alfa-beta/metabolismo , Subgrupos de Linfocitos T/efectos de los fármacos , Subgrupos de Linfocitos T/inmunología , Subgrupos de Linfocitos T/patología , Toxoplasma/patogenicidad , Toxoplasmosis/fisiopatología , Toxoplasmosis/prevención & control
13.
Genes Immun ; 12(6): 445-56, 2011 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-21451557

RESUMEN

The character of monocytes is both molded by and contributes to ongoing immune responses. We hypothesized that monocyte polarization could have durable qualities and these would be mediated partly by changes in the chromatin. We defined genome-wide expression and histone H4 acetylation (H4ac) changes after γ-interferon (IFN), α-IFN and interleukin-4 treatment. To identify genes with altered potential for expression, we stimulated polarized monocytes and identified genes up- or downregulated after polarization and stimulation but not either treatment alone. We also defined durability after an 18-h or 3-day washout. Genes uniquely regulated after the combination of polarization and stimulus were durably altered, with 51% of the effects being durable. This gene set was highly enriched for cytokine-induced alterations in H4ac, with P-values ranging from 10(-24) to 10(-37). Certain regulons defined by patterns of expression were also associated with altered H4ac, with P-values ranging from 10(-4) to 10(-29). Networking software revealed a high density of mitogen-activated protein (MAP) kinase nodes in these clusters. Therefore, some changes in monocyte gene expression were sustained over a 3-day period. These durably altered gene sets were enriched for changes in H4ac and were associated with potential MAP kinase effects.


Asunto(s)
Polaridad Celular , Histonas/metabolismo , Macrófagos , Monocitos/metabolismo , Acetilación , Células Cultivadas , Cromatina/genética , Expresión Génica , Histonas/química , Humanos , Interferón-alfa/farmacología , Interferón gamma/farmacología , Interleucina-4/farmacología , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Macrófagos/metabolismo , Proteínas Quinasas Activadas por Mitógenos/genética , Monocitos/citología , Monocitos/efectos de los fármacos , Monocitos/inmunología
14.
J Biotechnol ; 150(3): 408-16, 2010 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-20869404

RESUMEN

Recently, a new alkaline protease named HP70 showing highest homology to extracellular serine proteases of Stenotrophomonas maltophilia and Xanthomonas campestris was found in the course of a metagenome screening for detergent proteases (Niehaus et al., submitted for publication). Attempts to efficiently express the enzyme in common expression hosts had failed. This study reports on the realization of overexpression in Escherichia coli after structural modification of HP70. Modelling of HP70 resulted in a two-domain structure, comprising the catalytic domain and a C-terminal domain which includes about 100 amino acids. On the basis of the modelled structure the enzyme was truncated by deletion of most of the C-terminal domain yielding HP70-C477. This structural modification allowed effective expression of active enzyme using E. coli BL21-Gold as the host. Specific activity of HP70-C477 determined with suc-L-Ala-L-Ala-L-Pro-L-Phe-p-nitroanilide as the substrate was 30 ± 5 U/mg compared to 8 ± 1 U/mg of the native enzyme. HP70-C477 was most active at 40°C and pH 7-11; these conditions are prerequisite for a potential application as detergent enzyme. Determination of kinetic parameters at 40°C and pH=9.5 resulted in K(M)=0.23 ± 0.01 mM and k(cat)=167.5 ± 3.6s(-1). MS-analysis of peptide fragments obtained from incubation of HP70 and HP70-C477 with insulin B indicated that the C-terminal domain influences the cleavage preferences of the enzyme. Washing experiments confirmed the high potential of HP70-C477 as detergent protease.


Asunto(s)
Detergentes/química , Escherichia coli/genética , Metagenoma , Proteínas Recombinantes/química , Serina Proteasas/química , Secuencia de Aminoácidos , Dominio Catalítico , Detergentes/metabolismo , Electroforesis en Gel de Poliacrilamida , Concentración de Iones de Hidrógeno , Modelos Moleculares , Anotación de Secuencia Molecular , Datos de Secuencia Molecular , Filogenia , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Alineación de Secuencia , Serina Proteasas/genética , Serina Proteasas/metabolismo , Espectrometría de Masa por Láser de Matriz Asistida de Ionización Desorción , Stenotrophomonas maltophilia , Temperatura , Xanthomonas campestris
15.
Br J Anaesth ; 105(5): 648-56, 2010 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-20729532

RESUMEN

BACKGROUND: Intraoperative monitoring of neuronal function is important in a variety of surgeries. The type of general anaesthetic used can affect the interpretation and quality of such recordings. Although the principal effects of general anaesthetics are synaptically mediated, the extent to which they affect excitability of the peripheral afferent nervous system is unclear. METHODS: Forty subjects were randomized in a stratified manner into two groups, anaesthetized with either propofol or sevoflurane. The threshold tracking technique (QTRAC(®)) was used to measure nerve excitability parameters of the sensory action potential of the median nerve before and after induction of general anaesthesia. RESULTS: Several parameters of peripheral sensory afferent nerve excitability changed after induction of general anaesthesia, which were similar for both propofol and sevoflurane. The maximum amplitude of the sensory nerve action potential decreased in both groups (propofol: 25.3%; sevoflurane: 29.5%; both P<0.01). The relative refractory period [mean (sd)] also decreased similarly in both groups [propofol: -0.6 (0.7) ms; sevoflurane: -0.3 (0.5) ms; both P<0.01]. Skin temperature at the stimulation site increased significantly in both groups [propofol: +1.2 (1.0)°C; sevoflurane: +1.7 (1.4)°C; both P<0.01]. CONCLUSIONS: Small changes in excitability of primary sensory afferents after the induction of anaesthesia with propofol or sevoflurane were detected. These effects, which were non-specific and are possibly explained by changes observed in temperature, demonstrate possible anaesthetic effects on intraoperative neuromonitoring.


Asunto(s)
Anestésicos Generales/farmacología , Axones/efectos de los fármacos , Neuronas Aferentes/efectos de los fármacos , Potenciales de Acción/efectos de los fármacos , Adolescente , Adulto , Anciano , Anestésicos por Inhalación/farmacología , Anestésicos Intravenosos/farmacología , Axones/fisiología , Femenino , Humanos , Masculino , Éteres Metílicos/farmacología , Persona de Mediana Edad , Monitoreo Intraoperatorio/métodos , Neuronas Aferentes/fisiología , Propofol/farmacología , Sevoflurano , Adulto Joven
16.
AJNR Am J Neuroradiol ; 31(8): 1508-11, 2010 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-20488903

RESUMEN

BACKGROUND AND PURPOSE: Imaging is a key element in the study of many rodent models of human diseases. The application of DSA has been limited in these studies in part because of a lack of a method that allows serial intra-arterial examinations to be performed during an extended period of time. It was our intent to develop and test a method for performing sequential arterial catheterizations and DSA in rats. MATERIALS AND METHODS: Using a transfemoral approach, we subjected 12 adult male Harvey rats to 3 sequential DSA examinations during a 6- to 8-week period. At each examination, 2 selective arterial catheterizations and a DSA were performed. Animals were monitored for ill effects, and images from the 3 examinations were compared for quality and the presence of any arterial injury. RESULTS: Ten of the 12 rats survived all 3 examinations. There were no adverse effects noted and no evidence of arterial injury from the examinations. CONCLUSIONS: With the technique described, it is possible to perform serial arterial catheterizations and DSA in rats. This technique will be useful as an adjunct in the use of rodents for the study of human diseases.


Asunto(s)
Angiografía de Substracción Digital/métodos , Cateterismo/métodos , Angiografía Cerebral/métodos , Angiografía de Substracción Digital/mortalidad , Animales , Cateterismo/mortalidad , Angiografía Cerebral/mortalidad , Circulación Cerebrovascular , Modelos Animales de Enfermedad , Arteria Femoral , Masculino , Morbilidad , Ratas , Ratas Endogámicas , Roedores
17.
Toxicology ; 268(3): 165-70, 2010 Feb 09.
Artículo en Inglés | MEDLINE | ID: mdl-20026217

RESUMEN

A wide range of substances have been recognized as sensitizing, either to the skin and/or to the respiratory tract. Many of these are useful materials, so to ensure that they can be used safely it is necessary to characterize the hazards and establish appropriate exposure limits. Under new EU legislation (REACH), there is a requirement to define a derived no effect level (DNEL). Where a DNEL cannot be established, e.g. for sensitizing substances, then a derived minimal effect level (DMEL) is recommended. For the bacterial and fungal enzymes which are well recognized respiratory sensitizers and have widespread use industrially as well as in a range of consumer products, a DMEL can be established by thorough retrospective review of occupational and consumer experience. In particular, setting the validated employee medical surveillance data against exposure records generated over an extended period of time is vital in informing the occupational DMEL. This experience shows that a long established limit of 60 ng/m(3) for pure enzyme protein has been a successful starting point for the definition of occupational health limits for sensitization in the detergent industry. Application to this of adjustment factors has limited sensitization induction, avoided any meaningful risk of the elicitation of symptoms with known enzymes and provided an appropriate level of security for new enzymes whose potency has not been fully characterized. For example, in the detergent industry, this has led to general use of occupational exposure limits 3-10 times lower than the 60 ng/m(3) starting point. In contrast, consumer exposure limits vary because the types of exposure themselves cover a wide range. The highest levels shown to be safe in use, 15 ng/m(3), are associated with laundry trigger sprays, but very much lower levels (e.g. 0.01 ng/m(3)) are commonly associated with other types of safe exposure. Consumer limits typically will lie between these values and depend on the actual exposure associated with product use.


Asunto(s)
Alérgenos/toxicidad , Enzimas/toxicidad , Legislación Médica/tendencias , Exposición Profesional/legislación & jurisprudencia , Exposición Profesional/normas , Hipersensibilidad Respiratoria/etiología , Alérgenos/análisis , Animales , Comportamiento del Consumidor , Enzimas/análisis , Unión Europea , Humanos , Hipersensibilidad Respiratoria/fisiopatología , Valores Limites del Umbral
18.
Genes Immun ; 11(2): 124-33, 2010 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-19710693

RESUMEN

Systemic lupus erythematosus (SLE) is a polygenic disorder affecting approximately 1 in 1000 adults. Recent data have implicated interferons (IFN) in the pathogenesis, and the expressions of many genes downstream of IFNs are regulated at the level of histone modifications. We examined H4 acetylation (H4ac) and gene expression in monocytes from patients with SLE to define alterations to the epigenome. Monocytes from 14 controls and 24 SLE patients were used for analysis by chromatin immunoprecipitation for H4ac and gene expression arrays. Primary monocytes treated with alpha-IFN were used as a comparator. Data were analyzed for concordance of H4ac and gene expression. Network analyses and transcription factor analyses were conducted to identify potential pathways. H4ac was significantly altered in monocytes from patients with SLE. In all, 63% of genes with increased H4ac had the potential for regulation by IFN regulatory factor (IRF)1. IRF1 binding sites were also upstream of nearly all genes with both increased H4ac and gene expression. alpha-IFN was a significant contributor to both expression and H4ac patterns, but the greatest concordance was seen in the enrichment of certain transcription factor binding sites upstream of genes with increased H4ac in SLE and genes with increased H4ac after alpha-IFN treatment.


Asunto(s)
Interferones/metabolismo , Lupus Eritematoso Sistémico/genética , Lupus Eritematoso Sistémico/metabolismo , Monocitos/metabolismo , Análisis de Secuencia por Matrices de Oligonucleótidos , Acetilación , Adulto , Expresión Génica , Humanos , Interferón-alfa/metabolismo , Unión Proteica/genética , Procesamiento Proteico-Postraduccional
19.
Appl Microbiol Biotechnol ; 81(5): 875-86, 2009 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-18787818

RESUMEN

In the course of a microbial screening of soil samples for new oxidases, different enrichment strategies were carried out. With choline as the only carbon source, a microorganism was isolated and identified as Arthrobacter nicotianae. From this strain, a gene coding for a choline oxidase was isolated from chromosomal DNA. This gene named codA was cloned in Escherichia coli BL21-Gold and the protein (An_CodA) heterologously overexpressed as a soluble intracellular protein of 59.1 kDa. Basic biochemical characterization of purified protein revealed a pH optimum of 7.4 and activity over a broad temperature range (15-70 degrees C). Specific activities were determined toward choline chloride (4.70 +/- 0.12 U/mg) and the synthetic analogs bis(2-hydroxyethyl)-dimethylammonium chloride (0.05 +/- 0.45 x 10(-2) U/mg) and tris-(2-hydroxyethyl)-methylammonium methylsulfate (0.01 +/- 0.12 x 10(-2) U/mg). With increasing number of oxidizable groups, a significant decrease in activity was noted. Determination of kinetic parameters in atmorspheric oxygen resulted in K (M) = 1.51 +/- 0.09 mM and V (max) = 42.73 +/- 0.42 mU/min for choline chloride and K (M) = 4.77 +/- 0.76 mM and V (max) = 48.40 +/- 2.88 mU/min for the reaction intermediate betaine aldehyde respectively. Nuclear magnetic resonance spectroscopic analysis of the products formed during the enzyme reaction with choline chloride showed that in vitro the intermediate betaine aldehyde exists also free in solution.


Asunto(s)
Oxidorreductasas de Alcohol/genética , Oxidorreductasas de Alcohol/metabolismo , Arthrobacter/enzimología , Oxidorreductasas de Alcohol/química , Arthrobacter/genética , Arthrobacter/aislamiento & purificación , Betaína/análogos & derivados , Betaína/metabolismo , Colina/metabolismo , Clonación Molecular , ADN Bacteriano/química , ADN Bacteriano/genética , Estabilidad de Enzimas , Escherichia coli/genética , Expresión Génica , Concentración de Iones de Hidrógeno , Cinética , Espectroscopía de Resonancia Magnética , Datos de Secuencia Molecular , Peso Molecular , Análisis de Secuencia de ADN , Microbiología del Suelo , Especificidad por Sustrato , Temperatura
20.
Acta Psychiatr Scand ; 117(3): 198-206, 2008 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-18081924

RESUMEN

OBJECTIVE: Based on the reported association between cytokines with depression and suicide, and evidence of increased markers of inflammation in the brain of suicide victims, the present study examined the expression of cytokines in the orbitofrontal cortex of suicide victims. METHOD: In a postmortem sample obtained from the Brodman area 11 of suicides (n = 34) and controls (n = 17), real-time RT-PCR was used to compare the expression of mRNA species for tumor necrosis factor-alpha (TNF-alpha), interleukin (IL)-1 beta, 4, 5, 6, and 13. RESULTS: Increased expression of IL-4 was found in women suicide victims and IL-13 in men suicide victims. Elevated but not significant cytokine expression was also observed for TNF-alpha in women suicide victims. CONCLUSION: To our knowledge, these results provide the first evidence of the presence of mRNA transcripts of type 2 T-helper cytokines in the human orbitofrontal cortex and their increased expression in the brain of suicides.


Asunto(s)
Citocinas/genética , Trastorno Depresivo/inmunología , Lóbulo Frontal/inmunología , Suicidio/psicología , Adulto , Anciano , Anciano de 80 o más Años , Causas de Muerte , Trastorno Depresivo/mortalidad , Trastorno Depresivo/patología , Femenino , Lóbulo Frontal/patología , Alemania , Humanos , Interleucina-13/genética , Interleucina-1beta/genética , Interleucina-4/genética , Interleucina-5/genética , Interleucina-6/genética , Masculino , Persona de Mediana Edad , ARN Mensajero/genética , Valores de Referencia , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Factores Sexuales , Suicidio/estadística & datos numéricos , Factor de Necrosis Tumoral alfa/genética
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