RESUMEN
BACKGROUND AND AIMS: Endoscopic submucosal dissection (ESD) enables the curative resection of early malignant lesions and is associated with reduced recurrence risk. Due to the lack of comprehensive ESD data in the West, the German ESD registry was set up to evaluate relevant outcomes of ESD. METHODS: The German ESD registry is a prospective uncontrolled multicenter study. During a 35-month period, 20 centers included 1000 ESDs of neoplastic lesions. The results were evaluated in terms of en bloc, R0, curative resection rates, and recurrence rate after a 3-month and 12-month follow-up. Additionally, participating centers were grouped into low-volume (≤20 ESDs/y), middle-volume (20-50/y), and high-volume centers (>50/y). A multivariate analysis investigating risk factors for noncurative resection was performed. RESULTS: Overall, en bloc, R0, and curative resection rates of 92.4% (95% confidence interval [CI], 0.90-0.94), 78.8% (95% CI, 0.76-0.81), and 72.3% (95% CI, 0.69-0.75) were achieved, respectively. The overall complication rate was 8.3% (95% CI, 0.067-0.102), whereas the recurrence rate after 12 months was 2.1%. High-volume centers had significantly higher en bloc, R0, curative resection rates, and recurrence rates and lower complication rates than middle- or low-volume centers. The lesion size, hybrid ESD, age, stage T1b carcinoma, and treatment outside high-volume centers were identified as risk factors for noncurative ESD. CONCLUSION: In Germany, ESD achieves excellent en bloc resection rates but only modest curative resection rates. ESD requires a high level of expertise, and results vary significantly depending on the center's yearly case volume.
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Neoplasias Colorrectales/cirugía , Resección Endoscópica de la Mucosa , Neoplasias Esofágicas/cirugía , Neoplasias Gástricas/cirugía , Adulto , Anciano , Anciano de 80 o más Años , Competencia Clínica , Neoplasias Colorrectales/economía , Neoplasias Colorrectales/patología , Resección Endoscópica de la Mucosa/efectos adversos , Resección Endoscópica de la Mucosa/economía , Resección Endoscópica de la Mucosa/tendencias , Neoplasias Esofágicas/economía , Neoplasias Esofágicas/patología , Femenino , Alemania , Costos de la Atención en Salud , Hospitales de Alto Volumen , Hospitales de Bajo Volumen , Humanos , Masculino , Persona de Mediana Edad , Recurrencia Local de Neoplasia , Estadificación de Neoplasias , Estudios Prospectivos , Indicadores de Calidad de la Atención de Salud , Sistema de Registros , Medición de Riesgo , Factores de Riesgo , Neoplasias Gástricas/economía , Neoplasias Gástricas/patología , Factores de Tiempo , Resultado del TratamientoRESUMEN
Omalizumab is an effective therapeutic humanized murine IgE antibody in many cases of primary systemic mast cell activation disease (MCAD). The present study should enable the clinician to recognize when treatment of MCAD with omalizumab is contraindicated because of the potential risk of severe serum sickness and to report our successful therapeutic strategy for such adverse event (AE). Our clinical observations, a review of the literature including the event reports in the FDA AE Reporting System, the European Medicines Agency Eudra-Vigilance databases (preferred search terms: omalizumab, Xolair®, and serum sickness) and information from the manufacturer's Novartis database were used. Omalizumab therapy may be more likely to cause serum sickness than previously thought. In patients with regular adrenal function, serum sickness can occur after 3 to 10 days which resolves after the antigen and circulating immune complexes are cleared. If the symptoms do not resolve within a week, injection of 20 to 40 mg of prednisolone on two consecutive days could be given. However, in MCAD patients whose adrenal cortical function is completely suppressed by exogenous glucocorticoid therapy, there is a high risk that serum sickness will be masked by the MCAD and evolve in a severe form with pronounced damage of organs and tissues, potentially leading to death. Therefore, before the application of the first omalizumab dose, it is important to ensure that the function of the adrenal cortex is not significantly limited so that any occurring type III allergy can be self-limiting.
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Insuficiencia Suprarrenal/complicaciones , Factores Inmunológicos/efectos adversos , Mastocitos/efectos de los fármacos , Mastocitosis/tratamiento farmacológico , Omalizumab/efectos adversos , Enfermedad del Suero/inducido químicamente , Contraindicaciones de los Medicamentos , Glucocorticoides/uso terapéutico , Humanos , Mastocitos/inmunología , Mastocitos/metabolismo , Mastocitosis/inmunología , Mastocitosis/metabolismo , Prednisolona/uso terapéutico , Medición de Riesgo , Factores de Riesgo , Enfermedad del Suero/sangre , Enfermedad del Suero/tratamiento farmacológico , Enfermedad del Suero/inmunologíaRESUMEN
BACKGROUND: Systemic mast cell activation disease (MCAD) is characterized by an increased and unregulated release of mast cell mediators which can evoke a multifaceted clinical picture often resembling irritable bowel syndrome or fibromyalgia. Because of the considerable prevalence (~ 17 %) of MCAD surgeons are frequently unwittingly confronted with MCAD patients in whom unexpected intraoperative and postoperative complications may occur. Therefore, knowledge of the particular requirements is of relevance for surgical treatment of MCAD patients. OBJECTIVE: The present paper outlines a concept of surgical treatment of MCAD patients based on the literature which is illustrated by a case report on emergency laparoscopic cholecystectomy. CONCLUSIONS: Due to the high prevalence of MCAD in the general population it can be assumed that the frequency in the surgical patient population is similar. If a patient has MCAD, specific characteristics should be taken into account in the surgical procedure to avoid increased operative and complication risks resulting from MCAD.
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Colecistectomía Laparoscópica , Urgencias Médicas , Complicaciones Intraoperatorias/diagnóstico , Leucemia de Mastocitos/diagnóstico , Mastocitosis Sistémica/diagnóstico , Complicaciones Posoperatorias/diagnóstico , Estudios Transversales , Diagnóstico Diferencial , Endoscopía Gastrointestinal , Humanos , Complicaciones Intraoperatorias/etiología , Complicaciones Intraoperatorias/prevención & control , Leucemia de Mastocitos/epidemiología , Leucemia de Mastocitos/etiología , Leucemia de Mastocitos/prevención & control , Masculino , Mastocitosis Sistémica/epidemiología , Mastocitosis Sistémica/etiología , Mastocitosis Sistémica/prevención & control , Persona de Mediana Edad , Atención Perioperativa/métodos , Complicaciones Posoperatorias/etiología , Complicaciones Posoperatorias/prevención & control , Medicamentos bajo Prescripción/efectos adversos , Factores de RiesgoRESUMEN
BACKGROUND: Gastrointestinal endoscopies are increasingly being carried out with sedation. All of the drugs used for sedation are associated with a certain risk of complications. Data currently available on sedation-associated morbidity and mortality rates are limited and in most cases have substantial methodological limitations. The aim of this study was to record severe sedation-associated complications in a large number of gastrointestinal endoscopies. METHODS: Data on severe sedation-associated complications were collected on a multicentre basis from prospectively recorded registries of complications in the participating hospitals (median documentation period 27 months, range 9 - 129 months). RESULTS: Data for 388,404 endoscopies from 15 departments were included in the study. Severe sedation-associated complications occurred in 57 patients (0.01 %). Forty-one percent of the complications and 50 % of all complications with a fatal outcome (10/20 patients) occurred during emergency endoscopies. In addition, it was found that 95 % of the complications and 100 % of all fatal complications affected patients in ASA class ≥ 3. CONCLUSIONS: Including nearly 400,000 endoscopies, this study represents the largest prospective, multicenter record of the complications of sedation worldwide. The analysis shows that sedation is carried out safely in gastrointestinal endoscopy. The morbidity and mortality rates are much lower than previously reported in the literature in similar groups of patients. Risk factors for the occurrence of serious complications include emergency examinations and patients in ASA class ≥ 3.
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Efectos Colaterales y Reacciones Adversas Relacionados con Medicamentos/mortalidad , Endoscopía Gastrointestinal/mortalidad , Hipnóticos y Sedantes/uso terapéutico , Sistema de Registros , Adulto , Anciano , Femenino , Alemania/epidemiología , Humanos , Incidencia , Masculino , Persona de Mediana Edad , Seguridad del Paciente , Estudios Prospectivos , Factores de Riesgo , Tasa de SupervivenciaRESUMEN
We present the case of a 76-year-old lady in whom the work-up for iron-deficiency anaemia resulted in the finding of a giant gastric polyp. The polyp could be completely removed endoscopically. The final histology showed the rare entity of a pyloric gland adenoma with focal transition into a well-differentiated adenocarcinoma. The patient is well after a follow-up of 12 months. Pyloric gland adenoma was first described in 1990. In spite of its benign histological appearance, a transition into adenocarcinoma has been reported in up to 30 % of the cases. Thus, although relatively rare, the gastroenterologist/endoscopist, as well as the pathologist should be aware of the entity of pyloric gland adenoma.
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Adenocarcinoma/patología , Adenocarcinoma/cirugía , Adenoma/patología , Adenoma/cirugía , Mucosa Gástrica/cirugía , Neoplasias Gástricas/patología , Neoplasias Gástricas/cirugía , Anciano , Transformación Celular Neoplásica/patología , Endoscopía/métodos , Femenino , Mucosa Gástrica/patología , Humanos , Resultado del TratamientoRESUMEN
BACKGROUND: Reactivation of chronic hepatitis B in HBsAg carriers is a well known complication of chemo?therapy. The clinical spectrum ranges from asymptomatic hepatitis to fatal hepatic failure. Although it impairs the prognosis of cancer treatment, it may be overlooked due to other possible causes of liver damage. CASE REPORT: The patient presented with acute liver failure after 6 cycles of rituximab, fludarabine, and cyclophosphamide for low grade non-hodgkin's lymphoma. Differential diagnoses were chemotherapy-induced liver failure, autoimmune hepatitis, phenprocoumon-induced liver failure and infiltration of the liver by lymphoma. Finally, reactivation of hepatitis B with a fibrosing cholestatic pattern was identified. CONCLUSION: This case reminds clinicians that patients receiving high-intensive chemotherapy or immunosuppressive therapy should be screened for HBsAg. HbsAg positive patients should obtain prophylactic antiviral therapy with lamivudine or another substance active against HBV.
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Anticuerpos Monoclonales/efectos adversos , Ciclofosfamida/efectos adversos , Hepatitis B Crónica/complicaciones , Fallo Hepático Agudo/etiología , Linfoma no Hodgkin/tratamiento farmacológico , Vidarabina/análogos & derivados , Anticuerpos Monoclonales de Origen Murino , Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Diagnóstico Diferencial , Resultado Fatal , Hepatitis B Crónica/diagnóstico , Hepatitis B Crónica/patología , Humanos , Hígado/patología , Fallo Hepático Agudo/patología , Masculino , Persona de Mediana Edad , Recurrencia , Rituximab , Vidarabina/efectos adversosRESUMEN
In hilar cholangiocarcinoma, only 20-30% of the patients are candidates for curative surgical resection, leaving the majority with merely palliative treatment options. Since the natural history of hilar cholangiocarcinoma is dominated by local complications rather than metastatic disease, local palliative treatment seems a reasonable option. Here, endoluminal photodynamic therapy has emerged as a promising treatment with several prospective observational studies and 2 prospective randomised studies published which included nearly 200 patients. With low complication rate and morbidity, PDT achieves an increased median survival as well as an increased quality of life even in patients with reduced performance status. Radiotherapy is an alternative local treatment option applied as brachytherapy, external beam radiotherapy or combined modality treatment. To date, however, sufficient data from controlled clinical trials are lacking, thus palliative radiotherapy has to be considered an experimental treatment option.
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Neoplasias de los Conductos Biliares/terapia , Conductos Biliares Intrahepáticos , Braquiterapia , Colangiocarcinoma/terapia , Fotoquimioterapia , Neoplasias de los Conductos Biliares/diagnóstico por imagen , Neoplasias de los Conductos Biliares/mortalidad , Neoplasias de los Conductos Biliares/radioterapia , Colangiocarcinoma/diagnóstico por imagen , Colangiocarcinoma/mortalidad , Colangiocarcinoma/radioterapia , Colangiopancreatografia Retrógrada Endoscópica , Estudios de Seguimiento , Conducto Hepático Común , Humanos , Tumor de Klatskin/mortalidad , Tumor de Klatskin/radioterapia , Tumor de Klatskin/terapia , Cuidados Paliativos , Fotoquimioterapia/métodos , Estudios Prospectivos , Calidad de Vida , Dosificación Radioterapéutica , Ensayos Clínicos Controlados Aleatorios como Asunto , Factores de Tiempo , Resultado del TratamientoAsunto(s)
Colon , Enfermedades del Colon/diagnóstico , Colonoscopía , Quistes/cirugía , Cuerpos Extraños/diagnóstico , Perforación Intestinal/diagnóstico , Tercer Ventrículo/cirugía , Derivación Ventriculoperitoneal/instrumentación , Anciano , Remoción de Dispositivos , Humanos , Masculino , Complicaciones Posoperatorias/diagnósticoRESUMEN
BACKGROUND AND STUDY AIMS: The diagnosis of bile duct cancer is hampered by the low sensitivity of intraductal brush cytology and forceps biopsy. In the present study real-time reverse transcription polymerase chain reaction (RT-PCR) assays for the detection of human aspartyl (asparaginyl) beta-hydroxylase (HAAH) and homeobox B7 (HoxB7) mRNA from intraductal brush cytology specimens were established. Both markers are overexpressed in biliary cancer cell lines and possibly involved in the pathogenesis of bile duct cancer. PATIENTS AND METHODS: RT-PCR assays were validated for detection limit, in-assay variability, and inter-assay variability. Target gene expression was determined in brush cytology specimens from 16 patients with biliary strictures (11 with histologically proven cholangiocarcinomas and five with benign biliary strictures). RESULTS: The assay was quick (about 3 h), highly sensitive (with detection limits between 3 and 106 molecules), and reproducible (maximum in-assay variability 10.3 %, maximum inter-assay variability 11.8 %). The sensitivity of routine brush cytology alone was 36 % (four of 11 cases), with 100 % specificity. A combination with detection of HoxB7 and HAAH mRNA increased the overall diagnostic sensitivity to 82 %. CONCLUSIONS: Detection of these markers using the RT-PCR assays from brush cytology specimens described here may prove to be a useful additional tool for the diagnosis of bile duct carcinoma.
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Neoplasias de los Conductos Biliares/genética , Conductos Biliares Intrahepáticos , Colangiocarcinoma/genética , Regulación Neoplásica de la Expresión Génica , Proteínas de Homeodominio/genética , Oxigenasas de Función Mixta/genética , ARN Mensajero/genética , Adulto , Anciano , Anciano de 80 o más Años , Neoplasias de los Conductos Biliares/metabolismo , Neoplasias de los Conductos Biliares/patología , Línea Celular Tumoral , Colangiocarcinoma/metabolismo , Colangiocarcinoma/patología , Femenino , Humanos , Masculino , Persona de Mediana Edad , Reproducibilidad de los Resultados , Estudios Retrospectivos , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
OBJECTIVE: To assess characteristics and outcome of emergency patients with acute malaria. PATIENTS AND METHODS: We retrospectively assessed the clinical and laboratory parameters of 137 consecutive patients (87 males, 50 females; median age 37 years, range 17 - 67 years) presenting with acute malaria to our tertiary care center between 1992 and 2002. RESULTS: Falciparum malaria was diagnosed in 116/137 and tertian malaria in 19/137 patients; a single patient was infected with both parasites while in another case the type of parasite remained unclear. Infections were acquired in Africa (121), Asia , and in the Americas . One traveler visited multiple continents. Only 36 % (50/137) of patients had used malaria chemoprophylaxis. 128/137 patients were treated as in-patients; 22 of these had to be treated on an intensive care unit. According to the criteria of the German Society of Tropical Medicine, 44/137 (32 %; 95 % confidence interval (CI): 25 - 40 %) patients suffered from complicated malaria. The overall mortality rate was 2/137 (1.5 %; 95 % CI: 0,4 - 5.2 %); the mortality rate of complicated malaria tropica was 2/44 (4,5 %; 95 % CI 1,3 - 15 %). Patients with complicated malaria were significantly older than those with uncomplicated malaria. Median length of hospital stay was 4 days in uncomplicated and 9 days in complicated cases. Based on costs of EUR 2500 per case, an attack rate of > 3 % in East African travelers and a cost of EUR 55 for a chemoprophylaxis with mefloquine, chemoprophylaxis is cost-effective. CONCLUSION: In our retrospective analysis, complicated malaria tropica was associated with older age. Although malaria causes considerable morbidity, the overall mortality from severe malaria is low. Reinforcement of chemoprophylaxis especially in travelers to Africa could reduce malaria cases and is cost-effective.
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Malaria Falciparum/epidemiología , Malaria Vivax/epidemiología , Malaria/epidemiología , Plasmodium ovale , Adolescente , Adulto , África , Factores de Edad , Anciano , Asia , América Central , Quimioprevención/economía , Análisis Costo-Beneficio , Servicios Médicos de Urgencia , Femenino , Alemania/epidemiología , Humanos , Tiempo de Internación , Malaria/tratamiento farmacológico , Malaria/mortalidad , Malaria/prevención & control , Malaria Falciparum/tratamiento farmacológico , Malaria Falciparum/mortalidad , Malaria Falciparum/prevención & control , Malaria Vivax/tratamiento farmacológico , Malaria Vivax/mortalidad , Malaria Vivax/prevención & control , Masculino , Persona de Mediana Edad , Estudios Retrospectivos , América del Sur , Viaje , Resultado del TratamientoRESUMEN
HISTORY AND CLINICAL FINDINGS: A 72-year-old man was admitted with diarrhea, loss of weight and anemia. The diarrhea started after antibiotic treatment of a pneumonia and persisted for 6 months at admission. Monoclonal gammopathy was found on external examination. INVESTIGATIONS AND DIAGNOSIS: The work-up yielded iron deficiency anemia, monoclonal gammopathy (IgG kappa) and elevated polyclonal IgA due to Gliadin- and endomysium-antibodies. Duodenal mucosa biopsies showed villous atrophy and increased intraepithelial lymphocytes. Celiac disease was diagnosed. Unexpectedly, mediastinal lymphomas were found and the concomitant diagnosis of Hodgkin's disease was made. TREATMENT AND COURSE: On gluten free diet all symptoms of malabsorption resolved. Therapy for the Hodgkin lymphoma with chemotherapy was initiated. As Bleomycin associated lung disease occurred during therapy, radiotherapy was not administered. A complete remission could be achieved. CONCLUSIONS: The association of celiac disease and malignancy is well known. The pathogenesis is not fully understood, but a correlation between the duration of gluten exposure and the rate of malignancy was found. Thus, the chronic immunologic stimulation might also have contributed to the development of Hodgkin's disease in our patient, which to date has been reported only anecdotally.
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Enfermedad Celíaca/complicaciones , Enfermedad de Hodgkin/etiología , Anciano , Anemia Ferropénica/complicaciones , Protocolos de Quimioterapia Combinada Antineoplásica/administración & dosificación , Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Bleomicina/administración & dosificación , Bleomicina/efectos adversos , Enfermedad Celíaca/diagnóstico , Enfermedad Celíaca/dietoterapia , Dacarbazina/administración & dosificación , Dacarbazina/efectos adversos , Diarrea/etiología , Doxorrubicina/administración & dosificación , Doxorrubicina/efectos adversos , Enfermedad de Hodgkin/diagnóstico , Enfermedad de Hodgkin/tratamiento farmacológico , Humanos , Masculino , Paraproteinemias/complicaciones , Fibrosis Pulmonar/inducido químicamente , Vinblastina/administración & dosificación , Vinblastina/efectos adversosRESUMEN
OBJECTIVES: The impact of intrahepatic messenger RNA (mRNA) levels of interferon gamma (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha) on the outcome of antiviral treatment of chronic hepatitis C was evaluated. METHODS: Semiquantitative mRNA determination was performed on 36 pretreatment liver biopsies by reverse transcription/competitive polymerase chain reaction. RESULTS: Sustained response (normal aminotransferase levels and negative hepatitis C virus [HCV] RNA for more than 6 months) was achieved in 13 patients, whereas 23 of 36 patients did not achieve sustained response (12 partial responders, 11 complete nonresponders). In sustained responders, pretreatment intrahepatic mRNA levels of IFN-gamma and TNF-alpha were lower than in nonsustained responders (IFN-gamma, 0.23 +/- 0.10 vs. 0.35 +/- 0.07, respectively; p = 0.024 and TNF-alpha, 1.2 +/- 0.7 vs. 2.3 +/- 1.4, respectively; p= 0.009); similarly, HCV viral load was lower in sustained responders than in nonresponders (663,424 +/- 756,389 copies/mL vs. 1,656,713 +/- 1,517,683 copies/mL, respectively; p = 0.037). In addition, TNF-alpha mRNA levels were correlated to HCV viral load and liver fibrosis scores. CONCLUSIONS: Higher intrahepatic mRNA levels of IFN-gamma and TNF-alpha may reflect interferon resistance of HCV strains and may contribute to tissue damage in patients refractory to antiviral treatment.
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Antivirales/uso terapéutico , Hepatitis C Crónica/tratamiento farmacológico , Interferón gamma/biosíntesis , Hígado/inmunología , Factor de Necrosis Tumoral alfa/biosíntesis , Adolescente , Adulto , Anciano , Quimioterapia Combinada , Hepacivirus/efectos de los fármacos , Hepacivirus/aislamiento & purificación , Hepacivirus/fisiología , Hepatitis C Crónica/fisiopatología , Hepatitis C Crónica/virología , Humanos , Interferón alfa-2 , Interferón-alfa/uso terapéutico , Interferón gamma/genética , Persona de Mediana Edad , ARN Mensajero/genética , ARN Mensajero/metabolismo , ARN Viral/sangre , Proteínas Recombinantes , Ribavirina/uso terapéutico , Resultado del Tratamiento , Factor de Necrosis Tumoral alfa/genética , Carga ViralRESUMEN
Interferon-alpha (IFN-alpha) has potent pro-inflammatory and anti-viral functions. It exerts its effects by inducing intracellular proteins such as MxA. To analyse the role of intrahepatic interferon activation, IFN-alpha and MxA expression was studied by immunohistochemistry in explant livers of 20 patients with fulminant hepatic failure (FHF), 41 patients with chronic liver disease (CLD), and ten normal controls (NCs). In NCs only small numbers of Kupffer cells, but no hepatocytes, showed IFN-alpha and MxA expression. In contrast, significantly enhanced numbers of IFN-alpha- and MxA-positive Kupffer cells, along with small numbers of MxA-positive and larger numbers of IFN-alpha-positive lymphocytes, were found in CLD and in FHF. MxA protein was also expressed on hepatocytes and bile ducts in the vicinity of IFN-alpha-positive inflammatory infiltrates (hepatocytes: NCs: 0%, CLD: 8%, FHF: 68%; bile ducts: NCs: 19%, CLD: 46%, FHF: 83%). A significant correlation was found between the numbers of IFN-alpha- and MxA-positive cells (r=0.67, p<0.001). Thus, large amounts of IFN-alpha are released in the livers of patients with FHF, which is likely to contribute to immune-mediated liver cell damage. Intrahepatic MxA expression corresponds to IFN-alpha produced particularly by infiltrating inflammatory cells, rather than by hepatocytes themselves.
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Antivirales/metabolismo , Proteínas de Unión al GTP , Interferón-alfa/metabolismo , Fallo Hepático/metabolismo , Proteínas/metabolismo , Conductos Biliares Intrahepáticos/metabolismo , Hepatitis Crónica/metabolismo , Hepatitis Viral Humana/metabolismo , Hepatocitos/metabolismo , Humanos , Técnicas para Inmunoenzimas , Macrófagos/metabolismo , Proteínas de Resistencia a MixovirusRESUMEN
A reverse transcription/real-time polymerase chain reaction (PCR) assay was established to semi-quantify the mRNA levels of the human C-C chemokines RANTES, MIP-1beta and MCP-1 relative to the housekeeping gene beta-actin. The assay showed a high sensitivity (below 60 cDNA molecules/10 microl reaction) and dynamic range (8 log units); both within-assay and inter-assay variability were below 0.06 log units and the accuracy was +/-0.06 log units for all four chemokines. Moreover, it is demonstrated that a multi-specific DNA fragment, which had previously been constructed for competitive PCR, can be used as a reliable external standard. This allows a direct semi-quantitative comparison of different chemokine mRNA levels and is a convenient alternative to the use of different sets of homologous external standards. The method was successfully applied to the semi-quantification of chemokines in human liver specimens and should be useful in further studies on steady state mRNA levels of C-C chemokines from low cell numbers or small tissue specimens.
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Quimiocinas CC/aislamiento & purificación , ARN Mensajero/aislamiento & purificación , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Quimiocina CCL2/genética , Quimiocina CCL4 , Quimiocina CCL5/genética , Quimiocinas CC/genética , Hepatitis C Crónica/genética , Hepatitis C Crónica/inmunología , Humanos , Hígado/química , Cirrosis Hepática Biliar/genética , Cirrosis Hepática Biliar/inmunología , Proteínas Inflamatorias de Macrófagos/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/normasRESUMEN
BACKGROUND: Dysregulation of cytokines has been implicated in the pathogenesis of HIV infection. Therefore, we determined tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), IL-4, IL-10, and interferon-gamma (IFN-gamma) mRNA and serum levels in HIV-infected patients under nonstimulated conditions. MATERIAL AND METHODS: Blood samples of 32 HIV-infected patients and 10 healthy HIV-negative controls were analyzed. Cytokine serum levels were quantified by enzyme-linked immunosorbent assay (ELISA). Cytokine mRNA levels were determined semiquantitatively by competitive reverse transcriptase polymerase chain reaction (RT-PCR) and expressed as ratios relative to those of beta-actin. RESULTS: Competitive RT-PCR was shown to be more sensitive than protein ELISA in analyzing cytokine production. We found a significant correlation between steady-state mRNA ratios and serum protein levels for TNF-alpha. Significantly higher cytokine mRNA ratios were found in those patients with IL-10 and IFN-gamma levels detectable by ELISA. Steady-state mRNA ratios of TNF-alpha, IL-4, and IL-10 were significantly increased in patients with highly replicative HIV-infection. Furthermore, elevated IL-4:IFN-gamma ratios were related to both high viral load and loss of CD4 cells. DISCUSSION: Determination of steady-state mRNA ratios by semiquantitative RT-PCR represents a sensitive method to analyze cytokines in peripheral blood of HIV-infected patients under nonstimulated conditions. The data obtained with this technique provide further evidence for a T(H)1 to T(H)2 cytokine shift with progressive HIV disease.