RESUMEN
OBJECTIVE: The aim of this study was to test for an analgesic effect of exercise during labor. PATIENTS AND METHODS: 50 women in labor exercised continuously with moderate intensity on a bicycle ergometer for 20 minutes. During rest and exercise, they rated their pain on a visual analogue scale (VAS). Venous blood was sampled for beta-endorphin, cortisol and catecholamines during rest and directly after exercise. RESULTS: 84 percent of the women perceived uterine contractions during exercise as less painful than at rest. 76.2 percent objectified the pain relief by a reduction in VAS units 1.67 +/- 1.01. Beta-endorphin levels were much higher after exercise than at rest (P < 0.001). During exercise the fetal heart rate rose slightly within the reference range. Uterine contractions were more frequent during and after exercise than at rest (P < 0.05). CONCLUSION: Exercising on a bicycle ergometer during labor seems to be safe for the fetus, a stimulus to uterine contractions, and a source of analgesia, possibly due to the release of beta-endorphin.
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Analgesia/métodos , Ejercicio Físico/fisiología , Dolor de Parto/terapia , Dimensión del Dolor , Puntaje de Apgar , Bromuro de Butilescopolamonio/administración & dosificación , Cardiotocografía , Prueba de Esfuerzo , Femenino , Humanos , Recién Nacido , Dolor de Parto/fisiopatología , Masculino , Oxitocina/administración & dosificación , Parasimpatolíticos/administración & dosificación , Embarazo , Resultado del Tratamiento , betaendorfina/sangreRESUMEN
INTRODUCTION: This study was designed to assess changes in biochemical markers of bone remodeling in early postmenopausal women receiving calcium supplementation. MATERIALS AND METHODS: In a randomized cross-over study of eighteen weeks duration, the effect of a 6-week calcium supplementation (1000 mg calcium carbonate) on biochemical markers of bone resorption (collagen type I cross-linked C- and N-telopeptides) and bone formation (osteocalcin, total and bone-specific alkaline phosphatase), and total serum calcium was assessed in 27 early postmenopausal women. RESULTS: While total serum calcium levels increased significantly due to calcium supplementation (p<0.05), biochemical markers of both bone resorption and formation remained virtually unchanged. CONCLUSION: In contrast to other investigations, there was no significant short-term effect of calcium supplementation on biochemical markers of either bone resorption or formation.
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Biomarcadores/metabolismo , Remodelación Ósea/efectos de los fármacos , Calcio/administración & dosificación , Suplementos Dietéticos , Posmenopausia/fisiología , Calcio/farmacología , Estudios Cruzados , Esquema de Medicación , Femenino , Humanos , Persona de Mediana Edad , Posmenopausia/metabolismoRESUMEN
INTRODUCTION: The effect of pregnancy upon the maternal skeleton is not fully understood. The information that has been gathered by recent studies is conflicting with regard to overall loss or gain of bone during pregnancy. The aim of the present longitudinal, controlled study, therefore, was to investigate the effect of pregnancy on lumbar spine, wrist, and hip bone mineral density, and to describe bone remodeling during pregnancy as indicated by biochemical markers of both bone resorption and formation. MATERIALS AND METHODS: Thirty healthy women (15 subjects seeking pregnancy and 15 non-pregnant controls) were studied. Bone mineral density (BMD) was measured by dual-energy x-ray absorptiometry before conception and within 2 weeks after parturition. Markers of bone resorption (urinary cross-linked type I collagen N-telopeptides, serum type I collagen C-telopeptides) and bone formation (total and bone specific alkaline phosphatase, osteocalcin), and total serum calcium were analyzed before, during (once in each trimester), and after pregnancy. RESULTS: During pregnancy, BMD decreased significantly by 3.4+/-4.1% at the lumbar spine and 4.3+/-3.9% at the trochanter, while there was a slight but significant increase in BMD at the proximal 1/3 of the forearm (1.3+/-1.9%). Total hip and femoral neck BMD did not change significantly, nor did total and ultradistal forearm BMD. Bone resorption increased during pregnancy with peak levels in the third trimester (N-telopeptides) or post partum (C-telopeptides), respectively. The increase in bone resorption was accompanied by a significant decrease in serum calcium in the third trimester. Markers of bone formation showed a biphasic pattern with decreases from baseline to the first (total and bone specific alkaline phosphatase) or second trimester (osteocalcin), respectively, followed by a significant increase in the third trimester and post partum. There was no change in any parameter in the control group throughout the study. CONCLUSION: In conclusion, pregnancy is characterized by high bone turnover with resorption preceding formation. During the first and second trimester bone remodeling is uncoupled. Serum calcium decreases as bone resorption peaks in late pregnancy. There are significant decreases in bone mineral density at sites rich in trabecular bone, such as the lumbar spine and the trochanter.
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Densidad Ósea/fisiología , Remodelación Ósea/fisiología , Adulto , Fosfatasa Alcalina/sangre , Peso al Nacer , Resorción Ósea , Calcio/sangre , Colágeno Tipo I , Estradiol/sangre , Femenino , Fémur , Cuello Femoral , Edad Gestacional , Humanos , Vértebras Lumbares , Osteocalcina/sangre , Fragmentos de Péptidos/sangre , Péptidos , Embarazo , Procolágeno/sangre , Análisis de Regresión , Aumento de PesoRESUMEN
The aim of the present study was to assess whether the induction of specific immune responses by vaccination with the murine monoclonal anti-idiotypic antibody ACA125, which imitates the tumor-associated antigen CA125, has a positive influence on the survival of patients with recurrent ovarian carcinoma. Forty-two patients with platinum-pretreated recurrences were included in a clinical Phase I/II trial of consolidation in third-line therapy. Patients initially received four immunizations with 2 mg of alum-precipitated anti-idiotype ACA125 every 2 weeks and then monthly applications. No serious allergic reactions could be detected within a maximal control period of 56 months. Hyperimmune sera of 27 of 42 patients (64.2%) showed increased concentrations of human antimouse antibodies. Specific anti-anti-idiotypic antibodies as a marker for induced immunity were detected in 28 of 42 patients (66.7%). The survival of the whole ACA125-treated collective of patients after a mean of 12.6 antibody applications was 14.9 +/- 12.9 months. The survival of patients with a positive immune response was 19.9 +/- 13.1 months in contrast with 5.3 +/- 4.3 months in those patients without detectable anti-CA125 immunity (P < 0.0001). According to these results, vaccination with a suitable anti-idiotypic antibody offers an effective way to induce specific immunity against a primarily nonimmunogenic tumor antigen such as CA125 and is associated with a positive impact on the survival of patients with recurrent ovarian cancer with few side effects, which warrants a Phase III trial for ovarian cancer patients after primary therapy.
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Anticuerpos Antiidiotipos/uso terapéutico , Inmunoterapia , Neoplasias Ováricas/terapia , Anticuerpos Antiidiotipos/efectos adversos , Antígeno Ca-125/inmunología , Femenino , Humanos , Inmunidad , Persona de Mediana Edad , Neoplasias Ováricas/inmunología , Neoplasias Ováricas/mortalidad , Cuidados Paliativos , Recurrencia , Tasa de Supervivencia , Células Tumorales CultivadasRESUMEN
The portable blood gas analyzer OPTI Critical Care Analyzer was evaluated in comparison to routine laboratory assays using heparinized blood samples of adults and newborns. Within-run imprecision studies were performed with native blood using tonometry to adjust blood gas concentrations. The results obtained show a very close agreement between the OPTI system and the comparison methods for all parameters tested: hemoglobin (y=1.00x-0.2 g/l, r=0.997, n=81), sodium (y=1.13x-18.7 mmol/l, r=0.951, n=79), potassium (y=1.03x-0.04 mmol/l, r=0.972, n=79), pH (y=1.03x-0.29, r=0.958, n=57), pCO2 (y=1.03x-1.14 mm Hg, r=0.988, n=57) and pO2 (y=1.07x-0.85 mm Hg, r=0.995, n=57). The coefficients of variation for the within-run imprecision were below 1.1% for sodium and hemoglobin, and below 2.6% for all other parameters, except for pCO(2) with coefficients of variation up to 3.6% at low calibration gas concentrations. Due to this analytical performance and its portability, the OPTI system is well suited for low to medium test frequencies and immediate use in emergency rooms, intensive care or surgery units.
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Análisis de los Gases de la Sangre/instrumentación , Cuidados Críticos , Equipos y Suministros/normas , Sistemas de Atención de Punto/normas , Adulto , Electrólitos/sangre , Hemoglobinas/análisis , Humanos , Concentración de Iones de Hidrógeno , Recién Nacido , Reproducibilidad de los ResultadosRESUMEN
This study documents values of biochemical markers of bone remodeling in 106 patients with breast cancer. Based on scintigraphic and radiological findings, patients were divided into 3 groups: 19 patients with bone metastases, 65 patients without bone metastases and normal bone scintigrams, and 22 patients with pathological, non-malignant findings on scintigraphy without proof of bone metastases. Urinary cross-linked type I collagen N-telopeptides (NTx) and serum cross-linked type I collagen C-telopeptides (ICTP) were assessed as markers of bone resorption. Bone alkaline phosphatase (BAP) was assessed as a marker of bone formation. All three markers were significantly higher in patients with bone metastases compared to both patients without skeletal recurrence and those with pathological, non-malignant scintigraphic findings (p < 0.01). There were no statistically significant differences between the latter two groups. The clinical sensitivity for diagnosing bone metastases was 44% for NTx, 65% for ICTP, and 26% for BAP, respectively. The clinical specificitiy for discriminating patients with bone disease from those without were 79%, 91%, and 92% for NTx, ICTP, and BAP, respectively. In conclusion, markers of bone remodeling are increased in patients with breast cancer metastatic to the skeleton. The sensitivity of the markers presented in this paper did not seem to be sufficient enough for early identification of patients with subclinical bone recurrence in a clinical practice setting.
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Neoplasias Óseas/secundario , Neoplasias de la Mama/fisiopatología , Colágeno/sangre , Colágeno/orina , Péptidos/sangre , Péptidos/orina , Anciano , Fosfatasa Alcalina/sangre , Neoplasias Óseas/diagnóstico , Neoplasias Óseas/fisiopatología , Remodelación Ósea , Colágeno Tipo I , Femenino , Humanos , Persona de Mediana EdadRESUMEN
Clinical observations suggest that flecainide might pass the placenta more easily than digoxin, and that its transfer is less disturbed in case of hydrops fetalis than that of digoxin. The purpose of the study was to compare the materno-fetal transplacental transfer of digoxin, flecainide, and amiodarone, another antiarrhythmic agent used in the treatment of fetal tachyarrhythmia, and to assess the effect of an elevated umbilical venous pressure (UVP) on the transfer rate. Isolated lobules of 16 human placentas were dually perfused after spontaneous delivery or caesarean section. The transplacental transfer (area under the curve in the maternal compartment [maternal AUC], area under the curve in the fetal compartment [fetal AUC], kinetic parameters) of digoxin, flecainide, and amiodarone was calculated after these drugs were added to the maternal circuit. In five experiments, the effect of increased UVP on the transplacental transfer rate was assessed by elevating the UVP by 10 cm H2O. Flecainide efflux out of the maternal compartment was significantly greater than that of digoxin (maternal AUC 57.4% +/- 5.1 %/min vs 73.9% +/- 1.5%/min), whereas the flecainide influx into the fetal circulation was smaller (fetal AUC 9.3% +/- 4.1%/min vs 11.5% +/- 2.0%/min). Only in 50% of the experiments were the smallest amounts of amiodarone detectable in the fetal compartment. An elevation of the UVP reduced the influx of digoxin and flecainide into the fetal compartment (fetal AUC) from 11.5% +/- 2.0%/min to 7.4% +/- 1.9%/min and from 9.3% +/- 4.1% to 4.7% +/- 1.4%/min, respectively. Materno-fetal transplacental transfer of digoxin, flecainide, and amiodarone decreases in this sequence. Fetal cardiac insufficiency accompanied by an elevation of the UVP might reduce the transplacental transfer of these drugs, although no significant difference could be found between the reduction of transfer of digoxin and flecainide.
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Amiodarona/farmacocinética , Antiarrítmicos/farmacocinética , Digoxina/farmacocinética , Flecainida/farmacocinética , Placenta/metabolismo , Venas Umbilicales/fisiología , Amiodarona/uso terapéutico , Antiarrítmicos/uso terapéutico , Área Bajo la Curva , Digoxina/uso terapéutico , Femenino , Enfermedades Fetales/tratamiento farmacológico , Flecainida/uso terapéutico , Humanos , Técnicas In Vitro , Placenta/irrigación sanguínea , Embarazo , Flujo Sanguíneo Regional , Taquicardia Supraventricular/tratamiento farmacológico , Presión VenosaRESUMEN
Vaccination with anti-idiotypic antibodies has been described as a promising concept for treatment of several malignant diseases. The murine monoclonal anti-idiotypic antibody ACA125 imitates a specific epitope of the tumor-associated antigen CA125 expressed by 80% of ovarian carcinomas. In the first clinical trial it could be shown that mAb ACA125 is able to elicit anti-anti-idiotypic antibodies (Ab3) with anti-CA125 specificity in patients with advanced ovarian cancer. In order to improve the capabilities of anti-idiotype vaccines we generated a genetically engineered single-chain fragment (scFv) ACA125 composed of heavy- and light-chain variable regions connected by a flexible linker. The antigenicity of scFv ACA125 was demonstrated by immunizing rats i.p. with scFv or complete mAb in complete/incomplete Freund's adjuvants (CFA/IFA) or precipitated by aluminium hydroxide. Negative control groups included applications of irrelevant mouse IgG or adjuvants alone. Anti-anti-idiotypic antibodies (Ab3) directed against the mAb ACA125 as well as specific anti-CA125 antibodies (Ab1') could be detected in all animals treated with scFv in CFA/IFA. Nevertheless, antibody titers were lower than when the complete mAb ACA 125 was used. Suprisingly, an increase of specificity could not be observed in scFv-immunized animals, which had been expected because of the lack of heavy- and light-chain constant regions that could raise rather unspecific anti-isotypic and anti-allotypic rat anti-(mouse Ig) antibodies (RAMA). In contrast, the RAMA responses detected in these rats were even stronger than those following immunization with complete mAb ACA125. In conclusion, the anti-idiotypic scFv ACA125 alone cannot improve the immunogenic features of the corresponding mAb, but provides a useful tool for the further development of genetic vaccines.
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Anticuerpos Antiidiotipos/inmunología , Anticuerpos Monoclonales/inmunología , Hidróxido de Aluminio/farmacología , Animales , Formación de Anticuerpos/inmunología , Especificidad de Anticuerpos , Relación Dosis-Respuesta Inmunológica , Electroforesis en Gel de Poliacrilamida , Escherichia coli/metabolismo , Femenino , Adyuvante de Freund/farmacología , Inmunoglobulina G/inmunología , Ratones , Neoplasias Ováricas/metabolismo , Ratas , Ratas Sprague-DawleyRESUMEN
Increased renal ammoniagenesis is thought to be a causative factor for renal hypertrophy which occurs in several disorders accompanied by metabolic acidosis. We studied the influence of ammonia on the polarized release of degradation products of endogenous proteins in LLC-PK(1) cells. Release of acid-soluble radioactivity to the extracellular space decreased under the influence of NH(4)Cl with a remarkable transient reduction of basolateral release. Electron microscopically NH(4)Cl-treated cells showed numerous enlarged lysosomes suggesting an accumulation of incompletely degraded cytoplasmatic material in the lysosomal compartment. We conclude that split products of lysosomal degradation of endogenous proteins are preferentially transported to the extracellular space via the basolateral plasma membrane.
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Cloruro de Amonio/farmacología , Células LLC-PK1/efectos de los fármacos , Animales , Células LLC-PK1/ultraestructura , Lisosomas/metabolismo , Lisosomas/ultraestructura , Microscopía Electrónica , Proteínas/metabolismo , PorcinosRESUMEN
The following recommendations were drawn up by the Working Group "Point-of-Care Testing" of the DGKC and DGLM which was set up in 1997. This first document from the Working Group sets out the principles which should be observed when introducing point-of-care testing. These general recommendations are to be followed by further specific recommendations on individual procedures or groups of procedures, e.g. blood glucose, electrolytes in whole blood, blood gases, coagulation, toxicology, quality control and others, which will be drawn up by experts at the suggestion of the Working Group.
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Hospitales/normas , Laboratorios/normas , Sistemas de Atención de Punto/normas , Análisis Costo-Beneficio , Humanos , Laboratorios/economía , Sistemas de Atención de Punto/economía , Control de CalidadRESUMEN
OBJECTIVE: To determine the effect of the embryo transfer (ET) procedure on serum concentration of oxytocin. STUDY DESIGN: Prospective clinical study of 10 women undergoing in vitro fertilization (IVF) treatment with ET in the Section of Reproductive Medicine and Endocrinology at the Department of Obstetrics and Gynecology, University of Bonn, Germany. Serial blood samples were collected in time intervals of 20 s during embryo transfer procedure and serum oxytocin concentration was measured. RESULTS: In the absence of tenaculum placement, none of the procedures associated with ET led to an increase in serum oxytocin concentration. When tenaculum placement was used, it was temporally (four out of five patients) associated with an elevation in oxytocin level, which remained elevated until of the end of ET procedure. CONCLUSION: Application of a cervical tenaculum during ET or possibly also during intra uterine insemination (IUI) procedure can stimulate the release of oxytocin in some patients.
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Transferencia de Embrión , Oxitocina/sangre , Adulto , Cateterismo , Femenino , Fertilización In Vitro , Humanos , Persona de Mediana Edad , Embarazo , Estudios Prospectivos , Instrumentos QuirúrgicosRESUMEN
Anti-idiotypic antibodies, which imitate a tumor-associated antigen by their variable region, offer an elegant method for the induction of a specific immune response, when used as a surrogate antigen for immunization. We generated anti-idiotypic antibodies imitating 2 different tumor-associated antigens. I. CA125 for ovarian carcinomas and II. 14C5, a tumor-associated cell substrate adhesion molecule on breast cancer cells, whereas the first approach could be introduced in a first clinical trial and the second was evaluated in an immunocompetent animal model. For the induction of an immune response against CA125, 18 patients with advanced ovarian cancer (n = 6) or heavily pretreated recurrences (n = 12) were immunized with the anti-idiotypic antibody MAb ACA125. Patients were treated with 2 mg anti-idiotype antibody every two weeks for 4 injections i.m. and then monthly. 12 of 18 patients demonstrated an anti-anti-idiotypic (Ab3) response, which was to a lower extent also directed against CA125 and 9 of 18 patients developed a CA125 specific cellular immune response by their peripheral blood lymphocytes. Based on this data a follow-up clinical trial in advanced ovarian cancer patients with minimal residual disease in an adjuvant approach after primary therapy was started to evaluate the effect of the immune response on the progression free survival. For immunotherapy of breast cancer, we generated a murine monoclonal anti-idiotypic antibody (MAb ACA14C5), which imitates a cell substrate adhesion molecule on breast cancer cells. The anti-idiotype was introduced in an immunocompetent animal to prove his capability on induction of an immune and tumor response. The results showed a highly significant difference in the tumor growth of the ACA14C5 treated group in contrast to the controls starting the immunization on day 6 after tumor cell application with 10 of 12 animals being cured from their tumor burden. Prophylactic immunization against the invasion antigen of breast cancer by anti-idiotypic antibodies showed protection against increasing tumor burden. However, in the situation of established tumors only minor responses could be detected. Vaccination with anti-idiotypic antibodies comprises an effective method for induction of a specific immune response against non-immunogenic tumor-associated antigens and should be therefore considered in immunological approaches to tumor therapy, where the primary structure and sequence of the antigen, e.g. CA125, is up to now not available.
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Anticuerpos Monoclonales/uso terapéutico , Neoplasias de la Mama/terapia , Idiotipos de Inmunoglobulinas/inmunología , Inmunoterapia , Neoplasias Ováricas/terapia , Anticuerpos Monoclonales/inmunología , Especificidad de Anticuerpos/inmunología , Antígenos de Carbohidratos Asociados a Tumores/inmunología , Neoplasias de la Mama/inmunología , Antígeno Ca-125/inmunología , Femenino , Humanos , Neoplasias Ováricas/inmunología , Resultado del TratamientoRESUMEN
In a first clinical trial, 45 patients with advanced ovarian carcinoma and recurrences were treated with the murine monoclonal anti-idiotypic antibody (Ab2) designated ACA125 for active immunotherapy. The monoclonal antibody (MAb) ACA125 mimics a specific epitope of the tumor-associated antigen CA125 expressed by most malignant ovarian tumors. Patients with CA125-positive tumors are immunologically tolerant to CA125, which could be overcome by the use of an anti-idiotypic antibody as a surrogate for the tumor antigen CA125. An immunological response to the anti-idiotype ACA125 in these patients was associated with a statistically significant survival prolongation. Humoral immunity to ACA125 was assessed by induction of anti-anti-idiotypic antibodies (Ab3) directed against CA125. Using flow cytometric detection methods we observe alterations of the intracellular cytokines IFN-gamma, IL-2, and IL-4 at the single-cell level during the course of immunization. There was a strong increase of intracellular IFN-gamma and IL-2 characteristic for a Th1 cell type immune response after treatment with ACA125. A delayed induction of Th2 type response, which promotes antibody-mediated immunity by B cells, could also be detected. The understanding of the kinetics of Th1 and Th2 responses could be important to improve treatment schedules for effective immunotherapy with anti-idiotype vaccines.
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Anticuerpos Antiidiotipos/uso terapéutico , Anticuerpos Antineoplásicos/inmunología , Antígeno Ca-125/inmunología , Vacunas contra el Cáncer/inmunología , Neoplasias Ováricas/inmunología , Neoplasias Ováricas/terapia , Anticuerpos Antiidiotipos/inmunología , Anticuerpos Antineoplásicos/sangre , Anticuerpos Antineoplásicos/uso terapéutico , Antígenos CD/metabolismo , Vacunas contra el Cáncer/uso terapéutico , Citocinas/biosíntesis , Femenino , Humanos , Inmunofenotipificación , Linfocitos/inmunología , Neoplasias Ováricas/mortalidadRESUMEN
An important prerequisite for a successful pregnancy is that the maternal immune system does not reject the fetus. Down-regulation of the T helper 1 (TH1) associated cellular immune response could therefore be essential. With flow cytometric techniques, we show on a single cell level that both CD4+ and CD8+ T cells from peripheral blood produce less TH1 cytokines (i.e. IFN-gamma and IL-2) and more TH2 cytokines (i.e. IL-4) during normal human pregnancy and shortly after delivery than during non-pregnancy. The TH1/TH2 cytokine ratio in T cells of women during pregnancy and after delivery was significantly decreased. In contrast the TH1/TH2 ratio was elevated to near normal in women with recurrent spontaneous abortions, indicating a marked shift towards TH1 immunity. Fas antigen (CD95) on T cells was significantly elevated during pregnancy and in the post-delivery phase whereas the intracellular expression of anti-apoptotic protein Bcl-2 remained unchanged. Nevertheless Fas-mediated apoptosis in T cells was markedly reduced during normal human pregnancy. We hypothesize that TH1 cells undergo predominantly Fas-mediated apoptosis during pregnancy as has been shown in some TH2-prone diseases (e.g. SLE, HIV) where an elevated Fas expression on peripheral T cells is observed. This could explain the exacerbated occurrence of TH2-associated diseases in pregnancy.
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Apoptosis , Regulación hacia Abajo , Embarazo/inmunología , Células TH1/fisiología , Células Th2/fisiología , Femenino , Humanos , Interferón gamma/biosíntesis , Interleucina-2/biosíntesis , Interleucina-4/biosíntesis , Recuento de Linfocitos , Proteínas Proto-Oncogénicas c-bcl-2/biosíntesis , Receptor fas/biosíntesisRESUMEN
Four portable analyzers, HemoCue B-Glucose (I), Accu-Check III (II), One-touch II (III), and Glucometer Elite (IV), with different measuring principles were tested for their suitability for measuring blood glucose in neonates. Precision of all instruments is satisfactory. In the analysis of capillary blood from newborns, two instruments show an excellent accuracy; however, the scatter of the results for instrument (II) is about 1.6 times greater than for instrument (I). The inaccuracy of instruments (III) and (IV) is not acceptable from a clinical point of view. All devices show an influence of hematocrit, the magnitude of which varies between 5% (I) and 12% (III) for every 10% change of hematocrit. Instruments II and IV show that temperature has a marked influence on the readings; the same is true for oxygen in instrument IV. In conclusion, only instrument (I) has met the requirements of accurate and precise blood glucose determinations in neonates.
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Análisis Químico de la Sangre/instrumentación , Glucemia/análisis , Recién Nacido/sangre , Análisis Químico de la Sangre/normas , Análisis Químico de la Sangre/estadística & datos numéricos , Capilares , Estudios de Evaluación como Asunto , Hematócrito , Humanos , Hipoglucemia/sangre , Hipoglucemia/diagnóstico , Oxígeno/sangre , Estándares de Referencia , Reproducibilidad de los ResultadosRESUMEN
We have generated an immunoglobulin G1 (IgG1) murine monoclonal anti-idiotype antibody (Ab2) designated ACA125, which mimics a specific epitope on the tumor-associated antigen CA125. This antigen is expressed by most of malignant ovarian tumors. Patients with CA125-positive tumors are immunologically tolerant to CA125. We used ACA125 as a surrogate for the tumor-associated antigen CA125 for vaccine therapy of 16 patients with advanced epithelial ovarian cancer or recurrences. Each of the patients received a minimum of 3 injections up to 19 injections of the complete anti-idiotype MAb ACA125 at a dosage of 2 mg per injection. Nine of 16 patients developed anti-anti-idiotypic (Ab3) responses to the ACA125. All 9 patients generated specific anti-CA125 antibody demonstrated by reactivity with purified CA125. Nine of 16 patients developed a CA125-specific cellular immune response by their peripheral blood lymphocytes (PBL) and 3 of 16 showed an increase in gamma-interferon concentrations accompanied by Ab3 responses. Toxicity was limited to abdominal pain in one case, which led to the withdrawal of further immunizations. The median progression free survival in those patients, who showed a specific immune response to the tumor-associated antigen CA125, was 11.0 +/- 5.6 months without any other therapy, in contrast to 8.0 +/- 4.2 months in the anti-anti-idiotype negative group. This is the first report of the induction of a specific active immunity to the tumor-associated antigen CA125 in patients with advanced ovarian cancer treated with an anti-idiotype antibody that "mimics" CA125. Patients showed the development of a specific humoral and cellular immune response to an otherwise nonimmunogenic tumor antigen. The immune responses in patients treated with this anti-idiotype vaccine, the low rate of side effects, and the improved time to progression after the induction of a specific immune response against the tumor-associated antigen CA125 justify follow-up clinical trials in advanced ovarian cancer patients with minimal residual disease in an adjuvant approach.
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Anticuerpos Antiidiotipos/inmunología , Antígeno Ca-125/inmunología , Vacunas contra el Cáncer/inmunología , Neoplasias Ováricas/inmunología , Animales , Anticuerpos Antiidiotipos/uso terapéutico , Formación de Anticuerpos , Vacunas contra el Cáncer/uso terapéutico , Progresión de la Enfermedad , Supervivencia sin Enfermedad , Femenino , Humanos , Interferón gamma/biosíntesis , Ratones , Ratones Endogámicos BALB C , Persona de Mediana Edad , Neoplasias Ováricas/terapiaRESUMEN
The F(ab')2 fragment of the murine monoclonal anti-idiotypic antibody ACA125 mimicking the tumor-associated antigen CA125 is used as a vaccine for the induction of an anti-tumoral immunity in patients with ovarian carcinoma. We tried to generate a single-chain fragment (ScFv) composed of ACA125 heavy- and light-chain variable domains connected by a polypeptide linker as an alternative to the corresponding F(ab')2 fragment. Heavy- and light-chain genes of antibody-producing mouse hybridoma cell line were amplified separately and assembled into a ScFv gene with linker DNA by the polymerase chain reaction (PCR). The ScFv gene was ligated into the phagemid vector pCANTAB5E, which allows the production of both phage-displayed and soluble ScFv. Transformed Escherichia coli TG1 cells were infected with M13K07 helper phage to yield recombinant phage, which display ScFv fragments as a g3p fusion protein on the surface of the filamentous phage M13. Recombinant phages could be selected by binding to the idiotypic antibody OC125 after one round of panning and directly used to reinfect E. coli TG1 cells. The E. coli nonsuppressor strain HB2151 was infected with an antigen-positive phage clone, previously screened by enzyme-linked immunosorbent assay (ELISA), to express soluble ScFv fragments. Functional soluble ScFv binding to the idiotypic antibody OC125 F(ab')2 could be detected in the bacterial periplasm by Western blot and ELISA. The variable heavy- and light-chain genes of the ACA125 ScFv fragment were further sequenced and compared with known antibody sequences.
Asunto(s)
Anticuerpos Antiidiotipos/inmunología , Antígeno Ca-125/inmunología , Vacunas contra el Cáncer/inmunología , Fragmentos de Inmunoglobulinas/inmunología , Secuencia de Aminoácidos , Animales , Bacteriófago M13/genética , Antígeno Ca-125/genética , Vacunas contra el Cáncer/genética , Vacunas contra el Cáncer/uso terapéutico , Clonación Molecular , Electroforesis en Gel de Agar , Ensayo de Inmunoadsorción Enzimática , Escherichia coli , Femenino , Hibridomas , Fragmentos Fab de Inmunoglobulinas/genética , Fragmentos Fab de Inmunoglobulinas/inmunología , Fragmentos de Inmunoglobulinas/biosíntesis , Fragmentos de Inmunoglobulinas/genética , Región Variable de Inmunoglobulina/genética , Región Variable de Inmunoglobulina/inmunología , Ratones , Datos de Secuencia Molecular , Neoplasias Ováricas/terapia , Proteínas Recombinantes/genética , Proteínas Recombinantes/inmunología , SolubilidadRESUMEN
Our aim was to evaluate the isolated placental lobule to study maternofetal transplacental digoxin transfer and accumulation in placental tissue in vitro. Digoxin passage across the isolated lobule of 10 human placentas was calculated from repeated fetal and maternal perfusate samples, and placental tissue digoxin concentrations were measured at the end of the experiments. To determine the degree of overlap of the fetal and the maternal circulation, the antipyrine clearance was used. Digoxin disappearance from the maternal circuit was not significantly affected by the degree of overlap. In contrast, the increase of digoxin in the fetal compartment was significantly higher in "well-perfused" placentas (antipyrine clearance > 1.60 ml/min; n = 5) than in "malperfused" placentas (antipyrine clearance < 1.50 ml/min; n = 5) (end-feto to initial maternal digoxin ratio 0.44 +/- 0.08 vs. 0.30 +/- 0.08; p < 0.05), whilst the accumulation in placental tissue was higher in the latter group (0.45 +/- 0.07 vs. 0.62 +/- 0.10 ng/mg protein; p < 0.05). We conclude that the isolated placental lobule is suitable to quantify transplacental digoxin transfer in vitro, but the diffusion characteristics of each preparation have to be considered.