Effects of felbamate on the pharmacokinetics of a low-dose combination oral contraceptive.

The effects of felbamate on the pharmacokinetics of a low-dose combination oral contraceptive containing 30 micrograms ethinyl estradiol and 75 micrograms gestodene were assessed in a randomized, double-blind, placebo-controlled parallel-group study in healthy premenopausal female volunteers established in a regimen of oral contraceptive use. They received either placebo or 2400 mg/day felbamate from midcycle (day 15) to midcycle (day 14) of two consecutive oral contraceptive cycles (months 1 and 2). Pharmacokinetic assessments of ethinyl estradiol and gestodene were performed on day 14 of both cycles. To determine whether ovulation occurred, plasma progesterone and urinary luteinizing hormone levels were measured, and diaries recording vaginal bleeding were kept. Felbamate treatment resulted in a significant 42% decrease in gestodene area under the plasma concentration-time curve (0 to 24 hours) (p = 0.018) compared with baseline, whereas a minor but not clinically relevant effect was observed on the pharmacokinetic parameters of ethinyl estradiol. There were no changes in the pharmacokinetics of ethinyl estradiol or gestodene after placebo treatment. No volunteer showed hormonal evidence of ovulation; however, one volunteer reported the onset of intermenstrual bleeding during felbamate treatment. Because of the effect of felbamate on the pharmacokinetics of gestodene and the report of intermenstrual bleeding, it is possible that the contraceptive efficacy of low-dose combination oral contraceptives may be adversely affected during felbamate treatment.

High frequencies of antigen-specific hybridomas: dependence on immunization parameters and prediction by spleen cell analysis.

Hybridomas producing antibodies against soluble antigens have in most cases been difficult to establish. After fusion of myeloma cells with spleen cells obtained from mice immunized with a soluble protein, hybridomas secreting specific antibodies have been observed to occur very rarely among non-specific hybridomas. We found that the frequency of specific hybridomas correlates directly with the increase over background of the frequency of blast and/or plasma cells in the spleen (measured by cell size analysis) after antigenic stimulation. High yields of specific hybridomas were obtained simply by following a novel immunization technique consisting of several conventional preimmunization courses followed by 4 very high doses of antigen in saline on each of the last 4 days before fusion.

Familial atrial fibrillation with fetal onset.

A woman presented during two pregnancies (at 25 and 23 weeks' gestation, respectively) because the fetuses had rapid, irregular tachycardia and hydrops. After maternal drug treatment and achievement of slower fetal heart rates, the hydrops gradually resolved. Both babies were born full term with continuing atrial fibrillation. In the first, an ectopic atrial rhythm was temporarily achieved during high dose flecainide treatment but, in the younger sibling, all medications and repeated cardioversions failed even temporarily to convert the atrial fibrillation with an almost isoelectric baseline in ECG to sinus rhythm. Good rate control has been achieved with digoxin in both patients. No infective, immunological, or structural cause was found in either case, and thus an inherited aetiology is probable.

A simple, rapid and large capacity ELISA for biologically active native and recombinant human IFN gamma.

A rapid and sensitive enzyme-immunoassay for native and recombinant human interferon gamma is described. The test is performed in one step at room temperature and is based on the sandwich principle. The IFN gamma preparation is distributed with horse radish peroxidase-labeled monoclonal antibody to IFN gamma in microtiter plates previously coated with a second mab against IFN gamma. The amount of the IFN gamma mab sandwich fixed in the microtiter plate wells is proportional to the color developed after the addition of peroxidase-specific substrate. The two mab's used in the test neutralize IFN gamma and are directed against the same epitope. For this reason they can only detect the biologically active dimeric form of IFN gamma. The IFN gamma-ELISA works in phosphate buffer as well as in tissue culture medium or human serum. As the assay is routinely performed in 2 hours, the limit of detection is 3 U/ml of IFN gamma (0.3 ng/ml). If the assay is performed in 16 hours, the limit of detection decreases to 0.5 U/ml IFN gamma (0.06 ng/ml). The conditions to preserve the activity of IFN gamma preparation as standard are discussed.

Determination of placental blood flow by external monitoring of 113In.

A simple and noninvasive technique was developed to evaluate the blood perfusion rate within human placenta. The method is based on monitoring the accumulation of the isotope 113mIn in the placental intervillous space using a single detector as a measuring device. The result was expressed as an accumulation index calculated from the tracer appearance curve. The In accumulation index was significantly lower in preeclamptic patients than in the controls, but with a marked overlapping. The In index was significantly correlated with the simultaneously recorded maternal minute volume, 24-hour estrogen excretion and birthweight. No correlation was found with the placental weight or Laakso's perfusion index. Uterine contractions induced a significant decrease in the In index. The authors conclude that the In accumulation index can be used as an objective measure of the placental blood flow.

Mellow and frenetic antecedent music during athletic performance of children, adults, and seniors.

Previous research has yielded a contradictory picture of the effects of music on athletic performance. While athletes frequently report using music while training or during or before an event, laboratory studies have generally not detected a beneficial effect of music. The influence of music, judged mellow and frenetic, played before exercise was assessed by measuring stationary bicycle mileage. 60 volunteers from three age groups (child, adult, and senior) and with two levels of prior activity (high and low) were subjects. Each participant received three randomized 2-min. exercise trials, each preceded by 1-min. exposure to mellow music, frenetic music, or white noise. Mileage in both music conditions was significantly higher than that during the white-noise control trial except among the senior subjects. No significant differences between frenetic and mellow music were noted.

Effect of the left lateral recumbent position compared with the supine and upright positions on placental blood flow in normal late pregnancy.

The placental blood flow was assessed by the 99mTc accumulation method in 10 normal pregnancies in the left lateral recumbent position accomplished by a 15 degree wedge and in the supine position. The postural change caused a 17% decrease in the mean placental accumulation rate, which was not statistically significant. Ten patients were moved from the left lateral recumbent position to the upright position, which caused a statistically significant 23% decrease in the mean accumulation rate. Other haemodynamic variables studied were the maternal heart rate and the systolic and diastolic blood pressures. The clinical significance of the haemodynamic changes produced by alterations in posture are briefly discussed.

Myocardial infarction in a parturient. A case report with emphasis on medication and management.

Myocardial infarction is rare in pregnancy. We report a 38-year-old woman with a history of smoking and pre-eclampsia in her previous pregnancy, who suffered a non-Q infarction at 18 weeks of gestation. Stenoses in the left coronary artery were verified angiographically. She delivered a healthy child vaginally under epidural anaesthesia.

Prenatal activity of a fetus with early-onset, severe spinal muscular atrophy.

A case of early-onset, severe spinal muscular atrophy is reported. Normal fetal breathing movement patterns and heart rate accelerations were observed in spite of the severe hypotonia evident at birth.

Phaeochromocytoma in pregnancy.

A case of phaeochromocytoma diagnosed and treated during pregnancy is presented. The diagnosis was made by the typical paroxysmal symptoms with high blood pressure and confirmed by elevated plasma and urine catecholamine levels and ultrasound. After a short-term blockade with alpha and beta adrenergic antagonists an almost 10 cm large tumour was successfully excised at caesarean section. The problems with treatment of phaeochromocytoma during pregnancy are also discussed.

Structure and location of a GABA-A receptor complex in the central nervous system.

GABA-gated chloride channels in the central nervous system contain a regulatory site, the benzodiazepine receptor, through which drugs can modulate the efficiency of GABAergic synaptic transmission and thereby affect the degree of anxiety, muscle tension, vigilance and convulsions. The biochemical analysis of the purified receptor complex with monoclonal antibodies shows a heterooligomeric composition of two glycosylated subunits (alpha, beta). The immunoprecipitated complex contains the binding sites for GABA, benzodiazepines and the convulsant TBPS. The receptor complex was located, immunocytochemically, in synapses of brain regions rich in GABAergic nerve terminals.

Possible interaction between oxcarbazepine and an oral contraceptive.

The effect of oxcarbazepine (OCBZ) on the kinetics of an oral contraceptive containing ethinyloestradiol (EE) and levonorgestrel (LNG) was investigated in 13 healthy female volunteers who had previously received the contraceptive for at least 3 months. After 15 days of the first study cycle, each subject received, in addition to the oral contraceptive, 300 mg OCBZ on day 16, 300 mg twice daily on day 17, and 300 mg three times daily from day 18 of the first cycle to day 18 of the next menstrual cycle. The area under the curve values for both EE and LNG decreased when OCBZ was given with the oral contraceptive (p = 0.006, analysis of variance). The results indicate that OCBZ, like most antiepileptic drugs (AEDs), decreases the bioavailability of EE and LNG, perhaps by affecting metabolism or protein binding.

Ultrasonic and magnetic resonance imaging of fetal sacrococcygeal teratoma.

OBJECTIVE: To evaluate ultrasonic and magnetic resonance imaging (MRI) and Doppler examination in fetal sacrococcygeal teratoma (SCT), in respect to the postnatal findings and histological type of the tumor. STUDY DESIGN: Nine pregnancies complicated by histologically mature fetal sacrococcygeal teratoma in four cases and by immature/malignant teratoma in five cases. Transabdominal ultrasonic imaging and Doppler velocity waveforms were recorded during the second or last trimester in all cases, first trimester ultrasound examination was carried out in six cases and last trimester MRI in five cases. These findings were compared with postnatal and operative findings of the children. RESULTS: Ultrasound examination did not reveal intrapelvic parts of SCT, but this was possible by MRI. Velocity waveforms of the tumor arteries were similar in all histological types and the resistance index varied from 60 to 70. The mean gestational age at antepartal diagnosis was 25.2 weeks. Large tumor size with relatively large proportion of solid components was often recognized in cases with malignant/immature histology. CONCLUSIONS: Antepartal MRI is useful for examination of fetal SCT, but reliable differentiation of mature and immature SCT is not possible antepartally.

Resolving GABAA/benzodiazepine receptors: cellular and subcellular localization in the CNS with monoclonal antibodies.

Monoclonal antibodies, raised against a purified GABAA/benzodiazepine receptor complex from bovine cerebral cortex, have been used to visualize the cellular and subcellular distribution of receptorlike immunoreactivity in the rat CNS, cat spinal cord, and bovine and postmortem human brain. Two different antibodies have been used for these studies; bd-17 recognizes the beta-subunit (Mr 55 kDa) in all the species tested, whereas bd-24 recognizes the alpha-subunit (Mr 50 kDa) of bovine and human but not rat and cat tissues. In bovine and human brain, both antibodies produced very similar staining patterns, indicating a homogeneous receptor composition, at least in the brain areas investigated. The general distribution and density of receptor antigenic sites in all tissues studied were very similar to that of benzodiazepine binding sites radiolabeled with 3H-Ro 15-1788 and of glutamate decarboxylase (GAD)-stained nerve terminals. The results demonstrate a very high receptor density (around neuronal cell bodies and processes or less discretely distributed) in the rat olfactory bulbs, cerebral cortex, ventral pallidum, islands of Calleja, globus pallidus, hippocampus, dentate gyrus, substantia nigra, geniculate nuclei, inferior colliculus, cerebellum, reticular formation, spinal cord, and retina. In contrast, no receptors could be detected in white matter, pineal, pituitary, adrenals, and superior cervical ganglia. Only among the cerebellar layers did we observe a conspicuous difference between the staining intensity and the radiolabeling. In bovine and postmortem human brain, e.g., hippocampus, dentate gyrus, cerebral cortex, and substantia nigra, the same close correlation between the immunohistochemical and radiohistochemical findings was observed. At the electron microscopic level, the immune reaction product in the rat substantia nigra and globus pallidus, for example, was localized to pre- and postsynaptic membranes of axodendritic and axosomatic synapses. Whether the presynaptic labeling represents GABA autoreceptors is discussed. In the near future, the monoclonal antibodies will be used in double-labeling experiments with GAD to identify those GABAergic projections that are modulated by benzodiazepine minor tranquillizers. Furthermore, they could also be used, in studies of postmortem human brain, to diagnose receptor dysfunction possibly associated with CNS disorders such as epilepsy.

A GABA/benzodiazepine receptor complex from bovine brain: purification, reconstitution and immunological characterization.

A GABA/benzodiazepine receptor complex was purified from bovine cerebral cortex. The receptor fraction displayed binding sites for benzodiazepines as well as high and low affinity binding sites for GABA which are characteristics of the membrane-bound receptor. Two monoclonal antibodies of which one was directed against the 50 kd and the other against the 55 kd subunit were used for immunoprecipitation studies. Both of them were shown to quantitatively precipitate the entire receptor population. These results indicate that the binding sites for benzodiazepines and GABA (high and low affinity sites) reside on the same receptor complex containing a mixture of 50 kd and 55 kd subunits. Reconstitution of the receptor in phospholipid vesicles was achieved.

The visualization of neuronal benzodiazepine receptors in the brain by autoradiography and immunohistochemistry.

Recent methodological improvements in receptor autoradiography have enabled the in vitro and in vivo binding of the benzodiazepines in the brain to be visualized and pharmacologically characterized with an anatomical resolution unattainable by biochemical radioligand binding assays. This approach, combined with computerized microdensitometry, can be used not only to map the distribution of benzodiazepine receptors in the brain but also to quantify their regional densities. Furthermore, immunohistochemical studies, using monoclonal antibodies directed against the solubilized and purified GABA/benzodiazepine receptor-ionophore complex, have revealed the distribution of antigenic sites on brain neurons and their processes. The brain regions of intense immunoreactivity are known to contain a high density of GABA-ergic efferents and neuronal-type benzodiazepine receptors. Current trends and prospects in this area of receptor research are briefly reviewed.

Monoclonal antibodies reveal structural homogeneity of gamma-aminobutyric acid/benzodiazepine receptors in different brain areas.

Monoclonal antibodies (mAb) against a gamma-aminobutyric acid/benzodiazepine receptor complex (GABAA/BZR) were produced by using spleen cells from a mouse immunized with GABAA/BZR purified from bovine cerebral cortex. The mAb, most of which were of the IgG1 isotype could be divided into four groups (I-IV) specifying different antigenic structures. On immunoblots, group I mAb recognized exclusively the Mr 55,000 beta-subunit, while groups II and IV mAb recognized the Mr 50,000 alpha-subunit of bovine GABAA/BZR. Three of the four groups of mAb (I, III, and IV) crossreacted with both human and rat GABAA/BZR with the same subunit specificity as in bovine brain; the fourth group (II) crossreacted with human but not with the rat receptor. The binding sites for benzodiazepines as well as the high and low affinity GABA sites reside on the same structural complex as shown by immunoprecipitation. Ligand binding to these sites was not inhibited by mAb. Since quantitative immunoprecipitation of GABAA/BZR was achieved with mAb selective for either the alpha- or beta-subunit, both subunits occur in each individual receptor complex. The pattern of immunoblot staining suggests that the smaller alpha-subunit is not a processing product of the larger beta-subunit. Both alpha- and beta-subunits were present in all brain areas and species tested (rat cerebral cortex, cerebellum, and hippocampus; bovine cerebral cortex and cerebellum; human cerebral cortex). This suggests a uniform subunit composition of the receptor throughout the brain in contrast to earlier evidence for a heterogeneous subunit composition based on photoaffinity labeling.