Immunological consolidation of ovarian carcinoma recurrences with monoclonal anti-idiotype antibody ACA125: immune responses and survival in palliative treatment. See The biology behind: K. A. Foon and M. Bhattacharya-Chatterjee, Are solid tumor anti-idiotype vaccines ready for prime time? Clin. Cancer Res., 7:1112-1115, 2001.

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.

Impact of antihypertensive therapy with isradipine and metoprolol on early markers of hypertensive nephropathy.

In a double-blind, randomized trial with 26 male white patients with essential hypertension in World Health Organization Stages I and II, we examined the impact of calcium entry blockade (5 to 10 mg/day isradipine, N = 14) and beta-blockade (100 to 200 mg/day metoprolol, N = 12) on early markers of hypertensive nephropathy before and after 7 weeks' treatment. Excretion of total protein, albumin, alpha 1-microglobuline, and N-acetyl-beta-glucosaminidase (NAG) were measured in the 24-h urine by radial immunodiffusion and fluorimetric method, respectively. Before therapy, 8 of 26 patients had microproteinuria (31%), six had microalbuminuria (22%), six had elevated urinary NAG activity (22%), and three had elevated alpha 1-microglobulin excretion (11%). In these subjects anti-hypertensive therapy led to a fall in proteinuria (296 +/- 56 v 127 +/- 116 mg/day, P less than .01), albuminuria (44 +/- 24 v 25 +/- 12 mg/day, P less than .05), and NAG excretion (45 +/- 22 v 28 +/- 5, P less than .05). The higher the pretreatment value, the greater the fall was in proteinuria (r = +0.55, P less than .01), albuminuria (r = 0.80, P less than .001), and NAG excretion (r = 0.60, P less than .01). We did not observe any significant difference in clinical characteristics, blood pressure, or urinary excretion of protein, albumin, or NAG between the two treatment groups, either before or after therapy. Thus, antihypertensive therapy reduced excretion of total protein, albumin, and NAG activity in hypertensive patients with elevated pretreatment values, potentially indicating reversal of early hypertensive nephropathy.(ABSTRACT TRUNCATED AT 250 WORDS)

Analytical performance of a portable critical care blood gas analyzer.

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.

Enzymatic determination of bilirubin in serum of newborns--any advantage over previous methods?

Using a Hitachi 705 Selective Analyzer at 25 degrees C, precision and accuracy of the enzymatic determination of total bilirubin (DRI-STAT) in serum of newborns was studied. Imprecision (day-to-day) was unacceptably high using daily calibration, but could be reduced by using a fixed factor. Accuracy was checked by analysis of 70 newborn sera with the 'Candidate Reference Method' and DRI-STAT. It could be improved using calibrators with reference method values. Haemoglobin interference was studied by addition of different derivatives of fetal haemoglobin (oxihaemoglobin, desoxihaemoglobin, haemiglobin) to pools of newborn sera and control samples. DRI-STAT is more susceptible to interference by oxihaemoglobin than other routine methods. No difference was found in the influence of oxhihaemoglobin and desoxihaemoglobin; haemiglobin interferes significantly less.

Digoxin transfer across the isolated placenta is influenced by maternal and fetal albumin concentrations.

The aim of this study was to evaluate the influence of different maternal and fetal albumin concentrations on the transplacental transfer and the placental tissue accumulation of digoxin. Digoxin passage across the isolated lobules of 15 human placentae was calculated from repeated fetal and maternal perfusate samples, and placental tissue digoxin concentrations were measured at the end of the experiments. Metildigoxin (Lanitop) was added to the maternal medium at a concentration of 5.70 +/- 0.73 ng mL-1, and maternal and fetal perfusate albumin (BSA) concentrations were kept equal either at a high concentration of 21 g L-1 (Group I; n = 5) or at a low concentration of 3 g L-1 (Group III; n = 5), or differed with a materno-fetal gradient of 21:3 g L-1 (Group II; n = 5). In the experiments with low maternal albumin concentrations (Group III), digoxin concentrations in the maternal circuit decreased to 3.56 ng mL-1, whereas digoxin concentrations in the fetal circuit reached 2.59 ng mL-1 over a 3-h period. With maternal BSA concentrations of 21 g L-1 (Group I and Group II), the decrease in digoxin concentration in the maternal circuit was lower (P < 0.05), and digoxin tissue concentrations at the end of the experiments were smaller (0.45 +/- 0.07 and 0.42 +/- 0.03 v. 0.82 +/- 0.32 ng mg-1 protein, Group I and Group II v. Group III respectively; P < 0.05). Comparing only those lobules with similar high concentrations of maternal protein, fetal BSA concentrations of 21 g L-1 resulted in a greater increase in digoxin concentrations in the fetal circuit (end-feto to initial-maternal digoxin concentrations of 0.44 +/- 0.08 v. 0.37 +/- 0.04 ng mg-1 protein (Group I v. Group II respectively), although this was not significant. The data suggest that maternal and fetal serum albumin concentrations may have an influence on transplacental digoxin transfer, and this should be considered when treating fetuses with cardiac disease transplacentally with glycosides.

Serum oxytocin concentration during embryo transfer procedure.

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.

A monoclonal antiidiotypic antibody ACA 125 mimicking the tumor-associated antigen CA 125 for immunotherapy of ovarian cancer.

A new concept of oncological immunotherapy comprises the attempt to trigger the immune system of the host into a response against tumor cells. Antiidiotypic antibodies bearing the internal image of an antigen expressed on the surface of the tumor seem to be most suited for this purpose. We have generated a murine antiidiotypic antibody (ACA 125) functionally imitating the tumor-associated antigen CA 125, which can be detected in about 80% of ovarian carcinomas. The hybridoma cell was adapted to serum-free medium and antibody was produced in a hollow fiber cell culture system (Technomouse). ACA 125 (Ab2) shows high affinity for the paratope of Ab1 (affinity constant: 2.3 x 10(9) liters/mol) and binding of Ab2 to Ab1 is completely inhibited by the nominal antigen. Application of F(ab')2 fragments of ACA 125 to rats lead to an anti-CA 125 immunity by production of IgG and IgM antiantiidiotypic antibodies (Ab3) that bind to both ACA 125 and CA 125. Furthermore the induction of a non-MHC-restricted cell-mediated cytotoxicity for human ovarian adenocarcinoma cell line NIH-OVCAR3 (expressing CA 125 on its surface) could be proved; additionally complement-dependent cytotoxicity (CDC) as well as an antibody-dependent cellular cytotoxicity (ADCC) was observed. Thus, monoclonal antiidiotypic antibody ACA 125 fulfills recent criteria for an antibody, which might be successful in immunotherapy using the anti-idiotypic network approach.

Production of a single-chain fragment of the murine anti-idiotypic antibody ACA125 as phage-displayed and soluble antibody by recombinant phage antibody technique.

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.

Evaluation of immunological responses in patients with ovarian cancer treated with the anti-idiotype vaccine ACA125 by determination of intracellular cytokines--a preliminary report.

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.

Immunological responses to the tumor-associated antigen CA125 in patients with advanced ovarian cancer induced by the murine monoclonal anti-idiotype vaccine ACA125.

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.

Endogenous digoxin-like immunoreactive factors (DLIF) as measured by the CEDIA digoxin assay and a fluorescence polarization immunoassay.

The sensitivity of a new homogeneous enzyme immunoassay for the determination of digoxin (CEDIA Digoxin assay) and a fluorescence polarization immunoassay (FPIA) to interference by digoxin-like immunoreactive factors (DLIF) was studied in sera from pregnant women, newborns, patients undergoing hemodialysis and patients with renal insufficiency, but without hemodialysis. None of the patients had been treated with digoxin or digitoxin. Cross-reactivity of DLIF in the CEDIA assay was generally lower than in the FPIA. Data on the distribution DLIF of values and method comparisons showed that sera of the four patient groups reacted in a completely different way in both assays, suggesting that the nature of DLIF in the four groups is not identical. Addition of digoxin to sera of patients not treated with this drug resulted in a reduction of the apparent DLIF concentration in the CEDIA assay and the FPIA. This shows that DLIF interference may be less pronounced in sera of patients undergoing digoxin therapy compared to untreated persons. Although the CEDIA assay is less sensitive to DLIF interference than the FPIA, further efforts are needed to reduce the extent of this interference.

[Fructosamine reference ranges for pregnant patients and children determined using an improved NBT method]. / Fructosamin-Referenzbereiche für Schwangere und Kinder bestimmt mit einer verbesserten NBT-Methode.

Reference values for fructosamine in pregnancy show a decrease with progressing pregnancy, which can be explained by pregnancy-associated hemodilution. A normalization to 7.0 g/dl total protein leads to values independent of gestational age. For children and adolescents age-dependency of the reference range is abolished if values are related to total protein. More plausible values are obtained in longitudinal profiles if fluctuations of protein concentration are taken into account.

Results of the multicenter evaluation of a novel homogeneous immunoassay for digoxin based on the cloned enzyme donor immunoassay technology.

A new CEDIA assay for the measurement of digoxin in serum on random access analyzers was evaluated by twelve laboratories in Europe and the United States. Studies on the analytical range, reproducibility, calibration stability, recovery in controls, interlaboratory comparability, comparability with routine methods, and the effect of various interfering factors have been performed and the results are presented in this paper. The analytical performance was comparable to that of routine methods provided the manual pipetting step for pre-incubation was performed with accurate pipettes. A major advantage of the CEDIA Digoxin assay in terms of convenience is the simple two-point calibration procedure. Moreover, digoxin can be determined within 15 minutes after receiving the samples on random access analyzers like Boehringer Mannheim/Hitachi analysis systems. Thus, the CEDIA Digoxin assay represents an attractive alternative to the measurement of digoxin on dedicated immunochemical assay systems.

[The analgesic effect of exercise during labor]. / Der analgetische Effekt von körperlicher Aktivität auf Wehen unter der Geburt.

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.

[Semiquantitative determination of glucose in urine with a new test kit]. / Halbquantitative Glucose-Bestimmung im Urin mit einem neuen Testbesteck.

A new testing set for diluting urine allows to determine glucose in urine with the Diabur-test strip up to a concentration of 8.0 g/dl. The dilution by means of a pipette is absolutel;y practicable at a distinguished precision; the results show an excellent correlatiopn with the quantitative hexokinase method. Of all possible interfering substances tested only ascorbic acid influenced the reaction which, however, interfered only when minor glucosuria was present.

[A new, sensitive haemagglutination inhibition test for the specific detection of human choriongonadotropine (HCG) in urine]. / Ein neuer, empfindlicher Hämagglutinationshemmtest zum spezifischen Nachweis von Humanchoriongonadotropin (HCG) im Urin.

The reliability and clinical usefulness of a new pregnancy tube test with a sensitivity of 125 IU HCG/l was tested on 1142 urine samples. No false-negative results and 0.6% false-positive results were observed; a crossreaction with LH was seen above 1400 IU LH/l. In all cases of ectopic pregnancies (n = 49) the test yielded a positive result. In daily urine samples, collected during the luteal phase of the cycle, of 10 women who became pregnant, the test gave a positive result 13.6 days (on the average) after the rise of the basal body temperature. Especially in acute gynaecological emergencies the test is of value, because of its sensitivity and practicability.

[Reference values for the concentrations of triiodothyronine, thyroxine and thyrotropin in the blood serum of euthyroid children. Method: luminescence-enhanced enzyme immunoassay]. / Referenzwerte für die Konzentration von Trijodthyronin, Thyroxin und Thyreotropin im Blutserum euthyreoter Kinder. Methode: Lumineszenz-verstärkter Enzymimmunoassay.

Concentrations of triiodothyronine, thyroxine and thyrotropin were determined using the luminescence enhanced enzyme immunoassay method in blood serum of 343 (male = 197, female = 146) euthyroid children. Beyond the newborn age no significant difference was found for the thyrotropin concentration. Compilation of the values resulted in a physiological thyrotropin concentration of 1.73 +/- 0.81 (1s) microU/ml for infants, children and juveniles. In opposition to these results the thyroxine and triiodothyronine concentration showed an age-specific dependence. The thyroxine and triiodothyronine concentrations (means +/- 1s) decreased continuously from the infant age (TT4: 182.1 +/- 37.7 ng/ml, 234.9 +/- 48.6 nmol/l, TT3: 1.77 +/- 0.57 ng/ml, 2.73 +/- 0.88 nmol/l) to the juvenile age (TT4: 77.9 +/- 14.3 ng/ml, 100.5 +/- 18.4 nmol/l, TT3: 1.25 +/- 0.31 ng/ml, 1.93 +/- 0.48 nmol/l). The luminescence enhanced enzyme immunoassay technique is suitable for use in the routine thyroid laboratory.