Identification of an AR Mutation-Negative Class of Androgen Insensitivity by Determining Endogenous AR Activity.

CONTEXT: Only approximately 85% of patients with a clinical diagnosis complete androgen insensitivity syndrome and less than 30% with partial androgen insensitivity syndrome can be explained by inactivating mutations in the androgen receptor (AR) gene. OBJECTIVE: The objective of the study was to clarify this discrepancy by in vitro determination of AR transcriptional activity in individuals with disorders of sex development (DSD) and male controls. DESIGN: Quantification of DHT-dependent transcriptional induction of the AR target gene apolipoprotein D (APOD) in cultured genital fibroblasts (GFs) (APOD assay) and next-generation sequencing of the complete coding and noncoding AR locus. SETTING: The study was conducted at a university hospital endocrine research laboratory. PATIENTS: GFs from 169 individuals were studied encompassing control males (n = 68), molecular defined DSD other than androgen insensitivity syndrome (AIS; n = 18), AR mutation-positive AIS (n = 37), and previously undiagnosed DSD including patients with a clinical suspicion of AIS (n = 46). INTERVENTION(S): There were no interventions. MAIN OUTCOME MEASURE(S): DHT-dependent APOD expression in cultured GF and AR mutation status in 169 individuals was measured. RESULTS: The APOD assay clearly separated control individuals (healthy males and molecular defined DSD patients other than AIS) from genetically proven AIS (cutoff < 2.3-fold APOD-induction; 100% sensitivity, 93.3% specificity, P < .0001). Of 46 DSD individuals with no AR mutation, 17 (37%) fell below the cutoff, indicating disrupted androgen signaling. CONCLUSIONS: AR mutation-positive AIS can be reliably identified by the APOD assay. Its combination with next-generation sequencing of the AR locus uncovered an AR mutation-negative, new class of androgen resistance, which we propose to name AIS type II. Our data support the existence of cellular components outside the AR affecting androgen signaling during sexual differentiation with high clinical relevance.

Efficiency of the OVATEC unit for estrus detection and calf sex control in beef cows.

Seventy five spring calving Gelbvieh and Angus cows were utilized over a three year period to evaluate the usefulness of the OVATEC intravaginal probe for indicating the onset of estrus and providing the possibility to influence the sex of the offspring by choosing a breeding time in relation to critical changes in cervical mucus conductivity. Cows were randomly assigned by breed each year into one of four treatments: (1) probed and inseminated when impedance values declined, creating conditions expected to favor X-bearing sperm and with it an increase in the conception of females (PF); (2) probed and inseminated when impedance values were rebounding, expected to favor Y-bearing sperms and with it an increase in the conception of males (PB); (3) standing estrus (AI); or (4) natural service by bull (NS). Cows grazed or were fed hay from tall fescue-legume pastures. Lutalyse was used to synchronize estrus in a two injection scheme. Vaginal probe readings were taken at first injection, second injection and every 12 h thereafter for 6 days. Visual observations for estrus were obtained for PF, PB and AI every 12 h postsecond injection. Rectal palpations of ovaries were obtained at standing heat in all but NS treatments. In cycling cows, probe readings increased prediction of estrus onset (P < 0.10) compared to visual observations and were similar (P > 0.95) to rectal palpations in all probed cows. PF cows delivered heifer calves at greater rates (P < 0.025) than all other treatments whereas PB cows delivered bulls at greater rates (P < 0.05) than all other treatments. Heifer to bull ratios were not different (P > 0.95) for AI or NS treatments. The results of this study indicate that the potential exists for increasing female offspring conceptions utilizing cervical mucus conductivity as a gauge for insemination times.

Harmonic system analysis of the algae Valonia utricularis: contribution of an electrogenic transport system to gain and phase-shift of the transfer function.

Cell membrane properties of the giant marine alga Valonia utricularis were measured in the frequency domain between 1 Hz and 10 MHz by harmonic system analysis. Harmonic analysis was performed by imposing a sinusoidal electrical voltage on the cell interior via an internal microelectrode. Gain and phase-shift of the resulting sinusoidal membrane voltage were measured over the whole frequency range with an internal voltage microelectrode. Bode plots of gain and phase-shift allowed the determination of the electrical parameters of the equivalent electronic circuits of the cell membrane of V. utricularis, which showed dynamic and passive properties dependent on the pH of the external aqueous solution. The dynamic components of the membrane impedance were caused by an electrogenic transport system for chloride described previously (Wang, J., G. Wehner, R. Benz, and U. Zimmermann. 1991. Biophys. J. 59:235-248). The kinetic and equilibrium parameters of the transport system could be evaluated from the fit of Bode plots of gain and phase-shift. The frequency domain technique revealed complete agreement of transport parameters with previously published results. The data demonstrate that an electrogenic transport system can be driven by an oscillating electric field.

Influence of external chloride concentration on the kinetics of mobile charges in the cell membrane of Valonia utricularis: Evidence for the existence of a chloride carrier.

Charge pulse relaxation studies were performed on cells of the giant marine alga Valonia utricularis. Two exponential voltage relaxations were recorded as found previously (Benz, R., and U. Zimmermann. 1983. Biophys. J. 43:13-26.). The parameters of the two exponential voltage decays were studied as a function of the chloride concentration in the artificial sea water. Replacement of external chloride by 2(N-morpholino)ethanesulfonate (Mes(-)) had a dramatic influence on the four relaxation parameters. This chloride dependence could not be satisfactorily explained by the simplified model used earlier. Accordingly, additional reaction steps had to be included in the model. Only two relaxation processes could be resolved under all experimental conditions. This means that the heterogeneous complexation reactions, k(R) (association), and k(D) (dissociation) were too fast to be resolved. Therefore a carrier model with equilibrium heterogeneous surface reactions was used to fit the experimental results. From the charge pulse data at different chloride concentrations the translocation rate constants of the free and complexed carriers, k(S) and k(AS), through the membrane, as well as the total surface concentration of carrier systems, N(0), could be evaluated. The results described here indicate that the cell membrane of Valonia utricularis contains an electrogenic transport system for chloride.

The use of combination cervical plexus block and general anesthesia for radical neck dissection in a patient with severe chronic obstructive pulmonary disease.

Patients with severe chronic obstructive pulmonary disease (COPD) who receive a general anesthetic are at increased risk for postoperative pulmonary complications; therefore, it is desirable to avoid or to limit the dose of general anesthesia in these patients. Regional anesthesia, or a combination of regional and light general anesthesia, is an ideal choice for achieving this goal. This case report demonstrates how we used cervical plexus blocks in combination with light levels of general anesthesia for radical neck dissection in a patient with severe COPD.

Biphasic voltage relaxation pattern observed in cells of Eremosphaera viridis after injection of charge-pulses of short duration: detection of tip clogging of intracellular microelectrodes by charge-pulse technique.

Charge pulse experiments performed on the peat-bog alga Eremosphaera viridis revealed an unusual voltage relaxation behaviour. Injection of charge pulses of 1 microseconds duration resulted in an immediate charging of the membranes (time constant of the order of 40 ns). Nevertheless, the potential-measuring microelectrode recorded an exponential increase in membrane voltage with a time constant of about 1.3 ms. The maximum voltage value was recorded after about 3 ms, followed by an exponential decay with a time constant of about 9.6 ms. This biphasic time course was independent of the amplitude of the injected charge and of the location of the impaled microelectrodes in the vacuole. Centrifuged cells in which the chloroplasts and the other organelles were pelleted in one part of the cells showed the same electrical response. Electrical breakdown of the cell membranes resulted in the disappearance of the biphasic voltage response. In this case only the decaying relaxation process could be recorded with a time constant of 3 ms. After resealing of the membranes the original biphasic relaxation response was restored. Increasing concentrations of KCl in the bathing medium reduced both time constants almost correspondingly. The experimental findings were evaluated with an electrical equivalent circuit. Theoretical analysis with reference to the experimental data suggested that the delayed voltage response of the potential-recording electrode resulted from a membrane seal across the tip of this electrode. The resistance of this seal was calculated to be about 400 M omega. The specific resistances and capacitances of tonoplast and plasmalemma membranes were calculated from the decaying part of the biphasic relaxation curves. The average values were found to be 2.58 omega.m2 and 5 mF.m-2. The investigations reported here suggest that charge pulse experiments can be generally used for the detection of membrane and cytoplasmic material clogging of the tip of intracellular microelectrodes, a problem with which most electrophysiologists are faced when interpreting data obtained from impaled microelectrodes.

[Development of a rational monitoring strategy for workers exposed to tetrachloroethylene]. / Gestaltung einer rationellen Uberwachungsstrategie exponierter Werktätiger gegenüber Tetrachlorethen.

A 5 year follow-up of 130 workers exposed to tetrachlorethylene (dry cleaning) was performed, including measuring of exposure conditions (personal dosimetry, exposure tests) and clinical investigations. No signs of hepatoxicity of tetrachlorethylene could be detected, but slight derangements of nerval functions may develop in long-term exposed workers. Nerve motor and sensory conduction velocity shows a tendency to diminution. Recommendations are derived for biological monitoring and health surveillance of workers exposed to tetrachlorethylene.

Crop volume of chickens as affected by body size, sex, and breed.

Single Comb White leghorn (SCWL) hens, broiler cockerels, and two weight classes of SCWL roosters were utilized to determine the relationships of breed, sex, and body size to crop volume. Crop volume was significantly different (P less than .05) among all types of birds examined. Significant differences (P less than .05) of crop volume per kilogram body weight occurred between birds of opposite sex within a breed and between birds of identical sex but differing breed. However, within a breed, birds of identical sex but differing body size exhibited similar crop volume (cc/kg body weight).