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1.
Gastric Cancer ; 24(4): 959-969, 2021 07.
Artículo en Inglés | MEDLINE | ID: mdl-33576929

RESUMEN

BACKGROUND: For many cancer resections, a hospital volume-outcome relationship exists. The data regarding gastric cancer resection-especially in the western hemisphere-are ambiguous. This study analyzes the impact of gastric cancer surgery caseload per hospital on postoperative mortality and failure to rescue in Germany. METHODS: All patients diagnosed with gastric cancer from 2009 to 2017 who underwent gastric resection were identified from nation-wide administrative data. Hospitals were grouped into five equal caseload quintiles (I-V in ascending caseload order). Postoperative deaths and failure to rescue were determined. RESULTS: Forty-six thousand one hundred eighty-seven patients were identified. There was a significant shift from partial resections in low-volume hospitals to more extended resections in high-volume centers. The overall in-house mortality rate was 6.2%. The crude in-hospital mortality rate ranged from 7.9% in quintile I to 4.4% in quintile V, with a significant trend between volume categories (p < 0.001). In the multivariable logistic regression analysis, quintile V hospitals (average of 29 interventions/year) had a risk-adjusted odds ratio of 0.50 (95% CI 0.39-0.65), compared to the baseline in-house mortality rate in quintile I (on average 1.5 interventions/year) (p < 0.001). In an analysis only evaluating hospitals with more than 30 resections per year mortality dropped below 4%. The overall postoperative complication rate was comparable between different volume quintiles, but failure to rescue (FtR) decreased significantly with increasing caseload. CONCLUSION: Patients who had gastric cancer surgery in hospitals with higher volume had better outcomes and a reduced failure to rescue rates for severe complications.


Asunto(s)
Fracaso de Rescate en Atención a la Salud/estadística & datos numéricos , Gastrectomía/mortalidad , Hospitales de Alto Volumen/estadística & datos numéricos , Hospitales de Bajo Volumen/estadística & datos numéricos , Neoplasias Gástricas/mortalidad , Anciano , Femenino , Alemania , Mortalidad Hospitalaria , Humanos , Masculino , Persona de Mediana Edad , Oportunidad Relativa , Complicaciones Posoperatorias/mortalidad , Estudios Retrospectivos , Neoplasias Gástricas/cirugía , Carga de Trabajo/estadística & datos numéricos
4.
Nat Genet ; 25(2): 195-200, 2000 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-10835636

RESUMEN

The role of the opioid system in controlling pain, reward and addiction is well established, but its role in regulating other emotional responses is poorly documented in pharmacology. The mu-, delta- and kappa- opioid receptors (encoded by Oprm, Oprd1 and Oprk1, respectively) mediate the biological activity of opioids. We have generated Oprd1-deficient mice and compared the behavioural responses of mice lacking Oprd1, Oprm (ref. 6) and Oprk1 (ref. 7) in several models of anxiety and depression. Our data show no detectable phenotype in Oprk1-/- mutants, suggesting that kappa-receptors do not have a role in this aspect of opioid function; opposing phenotypes in Oprm-/- and Oprd1-/- mutants which contrasts with the classical notion of similar activities of mu- and delta-receptors; and consistent anxiogenic- and depressive-like responses in Oprd1-/- mice, indicating that delta-receptor activity contributes to improvement of mood states. We conclude that the Oprd1-encoded receptor, which has been proposed to be a promising target for the clinical management of pain, should also be considered in the treatment of drug addiction and other mood-related disorders.


Asunto(s)
Ansiedad/metabolismo , Depresión/metabolismo , Eliminación de Gen , Receptores Opioides delta/metabolismo , Receptores Opioides mu/metabolismo , Animales , Ansiedad/genética , Sitios de Unión , Oscuridad , Depresión/genética , Electrochoque , Femenino , Luz , Masculino , Ratones , Ratones Noqueados , Actividad Motora/efectos de los fármacos , Naloxona/farmacología , Naltrexona/análogos & derivados , Naltrexona/metabolismo , Naltrexona/farmacología , Antagonistas de Narcóticos/metabolismo , Antagonistas de Narcóticos/farmacología , Umbral del Dolor/efectos de los fármacos , Fenotipo , Receptores Opioides delta/deficiencia , Receptores Opioides delta/genética , Receptores Opioides kappa/deficiencia , Receptores Opioides kappa/genética , Receptores Opioides kappa/metabolismo , Receptores Opioides mu/deficiencia , Receptores Opioides mu/genética , Caracteres Sexuales , Natación
5.
Artículo en Inglés | MEDLINE | ID: mdl-22315630

RESUMEN

Background. Anthroposophic medicine is one of the widely used approaches of complementary and alternative medicine. However, few prospective studies have generated safety data on its use. Objectives. We aimed to assess adverse drug reactions (ADRs) caused by anthroposophical medicines (AMEDs) in the anthroposophical Community Hospital Havelhoehe, GERMANY. Study Design and Methods. Between May and November 2007, patients of six medical wards were prospectively assessed for ADRs. Suspected ADRs occurring during hospitalization were documented and classified in terms of organ manifestation (WHO SOC-code), causality (according to the Uppsala Monitoring Centre WHO criteria), and severity. Only those ADRs with a severity of grade 2 and higher according to the CTCAE classification system are described here. Results. Of the 3,813 patients hospitalized, 174 patients (4.6%) experienced 211 ADRs (CTCAE grade 2/3 n = 191, 90.5%, CTCAE grade 4/5 n = 20, 9.5%) of which 57 ADRs (27.0%) were serious. The median age of patients with ADRs (62.1% females) was 72.0 (IQR: 61.0; 80.0). Six patients (0.2%) experienced six ADRs (2.8% of ADRs) caused by eight suspected AMEDs, all of which were mild reactions (grade 2). Conclusion. Our data show that ADRs caused by AMEDs occur rarely and are limited to mild symptoms.

6.
J Obstet Gynaecol ; 32(4): 342-6, 2012 May.
Artículo en Inglés | MEDLINE | ID: mdl-22519477

RESUMEN

The present survey aims to identify predictors associated with the use of epidural analgesia (EA). Therefore, from October 2007 to June 2008, a survey was conducted in 193 pregnant women (mean age 31.7 years (SD 4.9); 64.8% primipara) attending a German general hospital with a specialisation in integrative medicine. Questionnaires, including Antonovsky's sense of coherence (SOC) were delivered antepartum. Delivery data were recorded within the hospital quality management programme. The adjusted odds ratio (OR) for EA use was significantly greater than one for women who had previously used EA (adjusted OR =4.1; CI: 1.03-16.31) and for the desire for a delivery without pain (adjusted OR =3.05; CI: 1.36-6.83). The likelihood of EA use decreased in multipara (adjusted OR =0.05; CI: 0.01-0.22). SOC was not found to be an independent predictor for EA use. However, women with high SOC more often preferred a delivery without EA (p for trend =0.037). In conclusion, first time labour, the desire for a delivery without pain and previous use of EA are independent predictors for the use of EA in labour. Further studies should clarify the predictive role of SOC in pregnancy.


Asunto(s)
Analgesia Epidural/estadística & datos numéricos , Parto Obstétrico/métodos , Dolor de Parto/terapia , Adulto , Femenino , Alemania , Humanos , Modelos Logísticos , Persona de Mediana Edad , Oportunidad Relativa , Paridad , Embarazo , Sentido de Coherencia , Encuestas y Cuestionarios , Adulto Joven
7.
Health Qual Life Outcomes ; 9: 85, 2011 Sep 30.
Artículo en Inglés | MEDLINE | ID: mdl-21961625

RESUMEN

BACKGROUND: Cancer Related Fatigue (CRF) and circadian rhythm have a great impact on the quality of life (HRQL) of patients with breast (BC) and colon cancer (CRC). Other patient related outcomes in oncology are measured by new instruments focusing on adaptive characteristics such as sense of coherence or self-regulation, which could be more appropriate as a prognostic tool than classical HRQL. The aim of this study was to assess the association of autonomic regulation (aR) and self-regulation (SR) with survival. METHODS: 146 cancer patients and 120 healthy controls took part in an initial evaluation in 2000/2001. At a median follow up of 5.9 years later, 62 of 95 BC, 17 of 51 CRC patients, and 85 of 117 healthy controls took part in the follow-up study. 41 participants had died. For the follow-up evaluation, participants were requested to complete the standardized aR and SR questionnaires. RESULTS: On average, cancer patients had survived for 10.1 years with the disease. Using a Cox proportional hazard regression with stepwise variables such as age, diagnosis group, Charlson co-morbidity index, body mass index (BMI)) aR and SR. SR were identified as independent parameters with potential prognostic relevance on survival While aR did not significantly influence survival, SR showed a positive and independent impact on survival (OR = 0.589; 95%-CI: 0.354 - 0.979). This positive effect persisted significantly in the sensitivity analysis of the subgroup of tumour patients and in the subscale 'Achieve satisfaction and well-being' and by tendency in the UICC stages nested for the different diagnoses groups. CONCLUSIONS: Self-regulation might be an independent prognostic factor for the survival of breast and colon carcinoma patients and merits further prospective studies.


Asunto(s)
Enfermedades del Sistema Nervioso Autónomo/psicología , Neoplasias de la Mama/psicología , Neoplasias del Colon/psicología , Calidad de Vida , Trastornos de Ansiedad/psicología , Estudios de Casos y Controles , Comorbilidad , Trastorno Depresivo/psicología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Observación , Pronóstico , Modelos de Riesgos Proporcionales , Estudios Prospectivos , Encuestas y Cuestionarios , Tasa de Supervivencia
8.
BJS Open ; 4(2): 310-319, 2020 04.
Artículo en Inglés | MEDLINE | ID: mdl-32207577

RESUMEN

BACKGROUND: The impact of hospital volume after rectal cancer surgery is seldom investigated. This study aimed to analyse the impact of annual rectal cancer surgery cases per hospital on postoperative mortality and failure to rescue. METHODS: All patients diagnosed with rectal cancer and who had a rectal resection procedure code from 2012 to 2015 were identified from nationwide administrative hospital data. Hospitals were grouped into five quintiles according to caseload. The absolute number of patients, postoperative deaths and failure to rescue (defined as in-hospital mortality after a documented postoperative complication) for severe postoperative complications were determined. RESULTS: Some 64 349 patients were identified. The overall in-house mortality rate was 3·9 per cent. The crude in-hospital mortality rate ranged from 5·3 per cent in very low-volume hospitals to 2·6 per cent in very high-volume centres, with a distinct trend between volume categories (P < 0·001). In multivariable logistic regression analysis using hospital volume as random effect, very high-volume hospitals (53 interventions/year) had a risk-adjusted odds ratio of 0·58 (95 per cent c.i. 0·47 to 0·73), compared with the baseline in-house mortality rate in very low-volume hospitals (6 interventions per year) (P < 0·001). The overall postoperative complication rate was comparable between different volume quintiles, but failure to rescue decreased significantly with increasing caseload (15·6 per cent after pulmonary embolism in the highest volume quintile versus 38 per cent in the lowest quintile; P = 0·010). CONCLUSION: Patients who had rectal cancer surgery in high-volume hospitals showed better outcomes and reduced failure to rescue rates for severe complications than those treated in low-volume hospitals.


ANTECEDENTES: El impacto del volumen hospitalario en los resultados de la cirugía del cáncer de recto ha sido poco investigado. Este estudio tuvo como objetivo analizar el impacto de los casos anuales de cirugía de cáncer de recto por hospital en la mortalidad postoperatoria (postoperative mortality, POM) y el fracaso en el rescate (failure to rescue, FtR). MÉTODOS: Todos los casos de pacientes hospitalizados con un diagnóstico de cáncer de recto y un código de procedimiento de resección rectal, tratados de 2012 a 2015, se identificaron a partir de datos hospitalarios administrativos a nivel nacional. Los hospitales se agruparon en cinco quintiles según el volumen de casos. Se determinó el número absoluto de pacientes, la POM y el FtR por complicaciones postoperatorias graves. El FtR se definió como la mortalidad hospitalaria después de una complicación postoperatoria documentada. RESULTADOS: Se identificaron 64.349 casos entre 2012 y 2015. La tasa de mortalidad hospitalaria global fue del 3,89% (n = 2.506). Las tasas brutas de mortalidad hospitalaria variaron de 5,34% (n = 687) en hospitales de muy bajo volumen a 2,63% (n = 337) en centros de muy alto volumen, con una tendencia distinta entre las categorías de centros (P < 0,001). En el análisis de regresión logística multivariante utilizando el volumen hospitalario como efecto aleatorio, los hospitales de muy alto volumen (53 intervenciones/año) tenían una razón de oportunidades (odds ratio, OR) ajustada por riesgo de 0,58 (i.c. del 95%: 0,47-0,73) en comparación con la tasa basal de mortalidad hospitalaria en hospitales de muy bajo volumen (6 intervenciones/año) (P < 0,001). La tasa global de complicaciones postoperatorias fue comparable entre los diferentes quintiles de volumen, pero el FtR disminuyó significativamente con el aumento del volumen de casos (15,63% FtR tras una embolia pulmonar en el quintil más alto versus 38,4% en el hospital del quintil más bajo, P = 0,01). CONCLUSIÓN: Los pacientes sometidos a cirugía de cáncer de recto en hospitales de gran volumen presentaron mejores resultados y una disminución de las tasas de fracaso en el rescate por complicaciones graves en comparación con los pacientes tratados en hospitales de bajo volumen.


Asunto(s)
Mortalidad Hospitalaria/tendencias , Neoplasias del Recto/mortalidad , Neoplasias del Recto/cirugía , Anciano , Anciano de 80 o más Años , Comorbilidad , Femenino , Alemania/epidemiología , Hospitales de Alto Volumen/estadística & datos numéricos , Hospitales de Bajo Volumen/estadística & datos numéricos , Humanos , Modelos Logísticos , Masculino , Persona de Mediana Edad , Evaluación de Resultado en la Atención de Salud , Neoplasias del Recto/epidemiología , Neoplasias del Recto/patología , Sistema de Registros , Estudios Retrospectivos
9.
BJS Open ; 3(5): 672-677, 2019 10.
Artículo en Inglés | MEDLINE | ID: mdl-31592096

RESUMEN

Background: Colonic cancer is the most common cancer of the gastrointestinal tract. The aim of this study was to determine mortality rates following colonic cancer resection and the effect of hospital caseload on in-hospital mortality in Germany. Methods: Patients admitted with a diagnosis of colonic cancer undergoing colonic resection from 2012 to 2015 were identified from a nationwide registry using procedure codes. The outcome measure was in-hospital mortality. Hospitals were ranked according to their caseload for colonic cancer resection, and patients were categorized into five subgroups on the basis of hospital volume. Results: Some 129 196 colonic cancer resections were reviewed. The overall in-house mortality rate was 5·8 per cent, ranging from 6·9 per cent (1775 of 25 657 patients) in very low-volume hospitals to 4·8 per cent (1239 of 25 825) in very high-volume centres (P < 0·001). In multivariable logistic regression analysis the risk-adjusted odds ratio for in-house mortality was 0·75 (95 per cent c.i. 0·66 to 0·84) in very high-volume hospitals performing a mean of 85·0 interventions per year, compared with that in very low-volume hospitals performing a mean of only 12·7 interventions annually, after adjustment for sex, age, co-morbidity, emergency procedures, prolonged mechanical ventilation and transfusion. Conclusion: In Germany, patients undergoing colonic cancer resections in high-volume hospitals had with improved outcomes compared with patients treated in low-volume hospitals.


Asunto(s)
Neoplasias del Colon/mortalidad , Neoplasias del Colon/cirugía , Mortalidad Hospitalaria/tendencias , Anciano , Anciano de 80 o más Años , Neoplasias del Colon/epidemiología , Neoplasias del Colon/patología , Comorbilidad , Femenino , Neoplasias Gastrointestinales/patología , Alemania/epidemiología , Hospitales de Alto Volumen/estadística & datos numéricos , Hospitales de Bajo Volumen/estadística & datos numéricos , Humanos , Masculino , Persona de Mediana Edad , Evaluación de Resultado en la Atención de Salud , Sistema de Registros
10.
Vet Parasitol ; 153(3-4): 329-37, 2008 May 31.
Artículo en Inglés | MEDLINE | ID: mdl-18359167

RESUMEN

Sarcoptic mange (or scabies) is an important skin disease which can affect a variety of species including humans, cattle, goats, sheep, horses, pigs, rabbits, and dogs. Approximately 300 million people are affected worldwide and in lifestock animals the infestation may lead to substantial economic losses caused by depression in growth and feed conversion rates. Diagnosis of Sarcoptes infestation is difficult and only a few serological tests have been developed using whole mite antigen for diagnosis of mange in animals. Here we describe the isolation and characterisation of cDNAs of several immunoreactive clones and their recombinant expression in Escherichia coli. Three of the proteins contain repetitive sequences which suggests that they might be involved in immune evasion. The application of these antigens in serodiagnosis and the suitability for diagnosis is discussed.


Asunto(s)
Antígenos/inmunología , Ensayo de Inmunoadsorción Enzimática/veterinaria , Sarcoptes scabiei/genética , Sarcoptes scabiei/inmunología , Escabiosis/veterinaria , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Clonación Molecular , ADN Complementario/química , ADN Complementario/genética , Ensayo de Inmunoadsorción Enzimática/métodos , Ensayo de Inmunoadsorción Enzimática/normas , Escherichia coli , Biblioteca de Genes , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa/métodos , Reacción en Cadena de la Polimerasa/veterinaria , Reproducibilidad de los Resultados , Escabiosis/diagnóstico , Sensibilidad y Especificidad , Alineación de Secuencia/veterinaria , Pruebas Serológicas/métodos , Pruebas Serológicas/normas , Pruebas Serológicas/veterinaria
11.
BMC Complement Altern Med ; 8: 26, 2008 Jun 05.
Artículo en Inglés | MEDLINE | ID: mdl-18533043

RESUMEN

BACKGROUND: To broaden the range of outcomes that we can measure for patients undergoing treatment for oncological and other chronic conditions, we aimed to validate a questionnaire measuring self-reported autonomic regulation (aR), i.e. to characterise a subject's autonomic functioning by questions on sleeping and waking, vertigo, morningness-eveningness, thermoregulation, perspiration, bowel movements and digestion. METHODS: We administered the questionnaire to 440 participants (female symbol: N = 316, male symbol: N = 124): 95 patients with breast cancer, 49 with colorectal cancer, 60 with diabetes mellitus, 39 with coronary heart disease, 28 with rheumatological conditions, 32 with Hashimoto's disease, 22 with multiple morbidities and 115 healthy people. We administered the questionnaire a second time to 50.2% of the participants. External convergence criteria included the German version of the Hospital Anxiety and Depression Scale (HADS-D), a short questionnaire on morningness-eveningness, the Herdecke Quality of Life Questionnaire (HLQ) and a short version questionnaire on self-regulation. RESULTS: A principal component analysis yielded a three dimensional 18-item inventory of aR. The subscales orthostatic-circulatory, rest/activity and digestive regulation had internal consistency (Cronbach-alpha: ralpha = 0.65 - 0.75) and test-retest reliability (rrt = 0.70 - 85). AR was negatively associated with anxiety, depression, and dysmenorrhoea but positively correlated to HLQ, self-regulation and in part to morningness (except digestive aR) (0.49 - 0.13, all p < 0.05). CONCLUSION: An internal validation of the long-version scale of aR yielded consistent relationships with health versus illness, quality of life and personality. Further studies are required to clarify the issues of external validity, clinical and physiological relevance.


Asunto(s)
Sistema Nervioso Autónomo/fisiología , Sistema Nervioso Autónomo/fisiopatología , Encuestas y Cuestionarios/normas , Adulto , Anciano , Anciano de 80 o más Años , Neoplasias de la Mama/fisiopatología , Enfermedad Crónica , Neoplasias Colorrectales/fisiopatología , Enfermedad Coronaria/fisiopatología , Estudios Transversales , Diabetes Mellitus/fisiopatología , Progresión de la Enfermedad , Femenino , Alemania/epidemiología , Enfermedad de Hashimoto/fisiopatología , Humanos , Masculino , Persona de Mediana Edad , Pronóstico , Calidad de Vida , Valores de Referencia , Reproducibilidad de los Resultados , Enfermedades Reumáticas/fisiopatología
13.
Mol Cell Biol ; 6(6): 2098-105, 1986 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-3023918

RESUMEN

HeLa cell nuclear extracts and wild-type or mutated simian virus 40 enhancer DNA were used in DNase I footprinting experiments to study the interaction of putative trans-acting factors with the multiple enhancer motifs. We show that these nuclear extracts contain proteins that bind to these motifs. Because point mutations which are detrimental to the activity of a particular enhancer motif in vivo specifically prevent protection of that motif against DNase I digestion in vivo, we suggest that the bound proteins correspond to trans-acting factors involved in enhancement of transcription. Using mutants in which the two domains A and B of the simian virus 40 enhancer are either separated by insertion of DNA fragments or inverted with respect to their natural orientation, we also demonstrate that the trans-acting factors bind independently to the two domains.


Asunto(s)
Proteínas de Unión al ADN/genética , Elementos de Facilitación Genéticos , Regulación de la Expresión Génica , Genes Reguladores , Virus 40 de los Simios/genética , Secuencia de Bases , Núcleo Celular/fisiología , Desoxirribonucleasa I
15.
Endocrinology ; 135(1): 476-9, 1994 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-8013388

RESUMEN

Although pleiotropic effects of glucocorticoids on the endocrine and exocrine pancreas are well documented, it remains controversial whether these effects are due to direct interactions of glucocorticoid hormones with their receptors in the respective target cells. We therefore examined gene expression of the glucocorticoid receptor (GR) in various functional compartments of the rat pancreas. Using in situ hybridization by generating a radioactive cRNA probe from a specific cDNA clone for the rat glucocorticoid receptor we found a gradient in the expression of receptor mRNA transcripts from endocrine > acinar > ductal cells. In contrast, no specific hybridization signal was observed in the endothelial cells of vascular structures. Using a monospecific polyclonal antibody against the rat glucocorticoid receptor in Western blot analyses we found significant expression of the 94 kD receptor protein in an enriched pancreatic acinar cell preparation. This data suggest that rat pancreatic exocrine cells might serve a direct target tissue for glucocorticoid action.


Asunto(s)
Expresión Génica , Islotes Pancreáticos/fisiología , Páncreas/fisiología , Receptores de Glucocorticoides/genética , Animales , Masculino , Páncreas/citología , Páncreas/metabolismo , ARN Mensajero/metabolismo , Ratas , Ratas Sprague-Dawley , Receptores de Glucocorticoides/metabolismo , Transcripción Genética
16.
Genes Brain Behav ; 2(2): 80-92, 2003 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-12884965

RESUMEN

In order to test the role of mu and kappa opioid receptors (Mu opioid receptor (MOR) and Kappa opioid receptor (KOR)) in hippocampal-dependent spatial learning, we analyzed genetically engineered null mutant mice missing the functional MOR or KOR gene. Compared to wild-type mice, the homozygous MOR null mutants exhibited an impairment in the ultimate level of spatial learning as shown in two distinct tasks, the 8-arm radial-maze and the Morris water-maze. Control behaviors were normal. The learning impairment could be associated with the impairment we found in the maintenance of long-term potentiation in mossy fibers in CA3. In comparison, there was no impairment in spatial learning in our KOR mutants or in mossy fibers (mf) in CA3 region long-term potentiation (LTP). Our work suggests that the MOR may play a positive role in learning and memory by increasing LTP in CA3 neurons.


Asunto(s)
Hipocampo/fisiología , Aprendizaje por Laberinto/fisiología , Receptores Opioides kappa/fisiología , Receptores Opioides mu/fisiología , Animales , Potenciación a Largo Plazo/fisiología , Masculino , Ratones , Ratones Noqueados , Fibras Musgosas del Hipocampo/fisiología
17.
Gene ; 32(1-2): 217-24, 1984 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-6099311

RESUMEN

An improved bacteriophage lambda cloning vector, lambda 2001, has been constructed. The phage includes a 34-bp polylinker oligonucleotide which introduces cleavage sites for XbaI, SstI, XhoI, EcoRI, HindIII and BamHI, and can accommodate 10-kb to 23-kb fragments. Inserts that destroy the BamHI or XhoI cloning sites may be recovered by excision at flanking sites in the polylinker sequence. Insertion of foreign DNA into lambda 2001 generates phage with a Spi- phenotype. The recombinant phage are able to grow on P2 lysogens but the parental vector phages are not. In the course of this work, the polylinker sequence was also introduced into M13mp8. This produced a new vector, M13mp12, with cloning sites for EcoRI, SmaI, XbaI, SstI, XhoI, BamHI, and HindIII.


Asunto(s)
Bacteriófago lambda/genética , Vectores Genéticos , Secuencia de Bases , Clonación Molecular , Enzimas de Restricción del ADN , ADN Recombinante , ADN Viral/genética , Oligonucleótidos/síntesis química
18.
J Immunol Methods ; 107(1): 59-66, 1988 Feb 24.
Artículo en Inglés | MEDLINE | ID: mdl-3343518

RESUMEN

50 fusion experiments were carried out to analyse heterohybridization efficiencies on mouse myeloma cells of the P3 X63 Ag8/653 line with human lymphocytes derived from peripheral blood, bone marrow, lymph node, spleen or synovial fluid. We found higher yields of growing and human Ig-producing hybridoma lines when lymphocytes from spleen or lymph node were fused. Although primary hybridomas could be established from fusions with bone marrow-derived cells, only in nine out of 1616 initially seeded wells was Ig production registered. Four fusions using immune cells from synovial fluid were made without success. Independently of the source of lymphocytes pokeweed mitogen (PWM) prestimulation had no enhancing effect on the percentage of wells with cell growth and this did not alter the IgM:IgG ratio in primary hybridomas (9:1), although cells from all compartments used here (with the exception of bone marrow cells) could be stimulated with PWM to produce both IgG and IgM in cultures. Cryopreserved lymphocytes from different sources could be used for fusions with comparable results registered for the fresh material.


Asunto(s)
Hibridomas/inmunología , Linfocitos/inmunología , Animales , Células de la Médula Ósea , Fusión Celular , Congelación , Humanos , Hibridomas/citología , Inmunoglobulina G/biosíntesis , Inmunoglobulina M/biosíntesis , Ganglios Linfáticos/citología , Activación de Linfocitos , Linfocitos/citología , Ratones , Mitógenos de Phytolacca americana/farmacología , Bazo/citología , Líquido Sinovial/citología
19.
J Neuroimmunol ; 81(1-2): 184-92, 1998 Jan.
Artículo en Inglés | MEDLINE | ID: mdl-9521621

RESUMEN

We have examined the distribution of the opioid receptor-like-1 (ORL-1) transcript in the human CNS as well as human immune cells by RT-PCR and RNAse protection. The hORL-1 mRNA was distributed throughout the brain and particularly abundant in cortical areas, striatum, thalamus and hypothalamus. In the immune system, gene transcription was observed in normal circulating lymphocytes and monocytes as well as in T, B and monocytic cell lines. A splice variant, lacking 15 nucleotides at the junction between exon 1 and exon 2, showed a distribution similar to the already known ORL-1 transcript. Altogether these results show comparable expression levels of the hORL-1 gene in both nervous and immune systems, suggesting that the ORL-1-encoded receptor may participate to neuronal and non-neuronal physiological functions in humans.


Asunto(s)
Proteínas del Tejido Nervioso/análisis , Péptidos Opioides/análisis , Adulto , Secuencia de Aminoácidos , Secuencia de Bases , Química Encefálica , Línea Celular , Clonación Molecular , Femenino , Humanos , Masculino , Persona de Mediana Edad , Datos de Secuencia Molecular , Proteínas del Tejido Nervioso/genética , Neuroinmunomodulación , Péptidos Opioides/genética , Especificidad de Órganos , Reacción en Cadena de la Polimerasa , Empalme del ARN , ARN Mensajero/análisis , Alineación de Secuencia , Homología de Secuencia , Nociceptina
20.
Neuroscience ; 106(4): 757-63, 2001.
Artículo en Inglés | MEDLINE | ID: mdl-11682161

RESUMEN

There is growing evidence that tonic activity of the opioid system may be important in the modulation of affective state. Naloxone produces a conditioned place aversion in rodents, an effect that is centrally mediated. Previous pharmacological data using antagonists with preferential actions at mu-, delta-, and kappa-opioid receptors indicate the importance of the mu-opioid receptor in mediating this effect. We sought to test the mu-opioid receptor selectivity of naloxone aversion using mu-opioid receptor knock-out mice. mu-Opioid receptor knock-out and wild-type mice were tested for naloxone (10 mg/kg, s.c.) aversion using a place conditioning paradigm. As a positive control for associative learning, knock-out mice were tested for conditioned place aversion to a kappa agonist, U50,488H (2 mg/kg, s.c.). Naloxone produced a significant place aversion in wild-type mice, but failed to have any effect in mu-opioid receptor knock-out mice. On the other hand, both knock-out and wild-type mice treated with U50,488H spent significantly less time in the drug-paired chamber compared to their respective vehicle controls. We conclude that the mu-opioid receptor is crucial for the acquisition of naloxone-induced conditioned place aversion. Furthermore, in a separate experiment using C57BL/6 mice, the delta-selective antagonist naltrindole (10 or 30 mg/kg, s.c.) failed to produce conditioned place aversion.Taken together, these data further support the notion that naloxone produces aversion by antagonizing tonic opioid activity at the mu-opioid receptor.


Asunto(s)
Reacción de Prevención/fisiología , Encéfalo/metabolismo , Condicionamiento Psicológico/fisiología , Naloxona/farmacología , Naltrexona/análogos & derivados , Antagonistas de Narcóticos/farmacología , Péptidos Opioides/metabolismo , Receptores Opioides mu/deficiencia , 3,4-Dicloro-N-metil-N-(2-(1-pirrolidinil)-ciclohexil)-bencenacetamida, (trans)-Isómero/farmacología , Analgésicos no Narcóticos/farmacología , Animales , Reacción de Prevención/efectos de los fármacos , Encéfalo/efectos de los fármacos , Condicionamiento Psicológico/efectos de los fármacos , Relación Dosis-Respuesta a Droga , Habituación Psicofisiológica/efectos de los fármacos , Habituación Psicofisiológica/fisiología , Masculino , Ratones , Ratones Endogámicos C57BL , Ratones Noqueados , Actividad Motora/efectos de los fármacos , Actividad Motora/fisiología , Naltrexona/farmacología , Neuronas/efectos de los fármacos , Neuronas/metabolismo , Péptidos Opioides/antagonistas & inhibidores , Fenotipo , Receptores Opioides delta/efectos de los fármacos , Receptores Opioides delta/metabolismo , Receptores Opioides kappa/efectos de los fármacos , Receptores Opioides kappa/metabolismo , Receptores Opioides mu/genética , Transmisión Sináptica/efectos de los fármacos , Transmisión Sináptica/fisiología
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