Differential changes in inhibin A, activin A, and total alpha-subunit levels in granulosa and thecal layers of developing preovulatory follicles in the chicken

Accumulating evidence implicates inhibins and activins as endocrine and local regulators of follicular development in mammals, and it was recently confirmed that inhibin/activin alpha and betaA genes are also expressed in the avian ovary. To investigate the potential involvement of these proteins in the chicken ovary, thecal and granulosa layers of the four largest follicles (F1-F4) and the most recent postovulatory follicle were collected from hens (10/group) killed 4, 12, and 20 h before the expected time of F1 ovulation. Inhibin A and activin A concentrations of tissue extracts (expressed per mg DNA) were measured using validated two-site enzyme-linked immunosorbent assays; total immunoreactive inhibin alpha-subunit (ir-alpha) was also measured by heterologous RIA (Monash assay). Inhibin A and ir-alpha were largely confined to the granulosa layer, whereas activin A was much more abundant in the thecal layer. Granulosa inhibin A contents were similar in F4 and F3, but increased approximately 40-fold from F3-F1 (P < 0.0001). As such, the F1 granulosa layer was by far the richest source of inhibin A in the chicken ovary, but contained very little activin A. Total ir-alpha in granulosa was much more abundant than inhibin A and increased only 3-fold from F4-F1 (P < 0.001). Activin A in both granulosa and theca showed little variation between F1 and F4 follicles (by ANOVA, P > 0.05). The inhibin A content of F1 granulosa was maximal 12 h before ovulation and had fallen approximately 6-fold (P < 0.0001) within 8 h, suggesting an inhibitory effect of the preovulatory LH surge on the F1 capacity to synthesize inhibin A. Inhibin A, activin A, and ir-alpha were all less in the postovulatory follicle compared with F1 before ovulation (P < 0.0001). In conclusion, application of the present two-site enzyme-linked immunosorbent assays to the chicken ovary revealed 1) divergent tissue distribution of inhibin A and activin A within preovulatory follicles, and 2) differential regulation of granulosa cell production of inhibin A and activin A dimers during preovulatory follicular development. These findings of dynamic changes in inhibin A, activin A, and total ir-alpha support the hypothesis that these proteins subserve regulatory roles during preovulatory follicular development in the hen.     

Atypical acute dystonia

Acute dystonia is a common adverse effect following anti-psychotic medication, which mainly appears shortly after beginning treatment or increasing the dosage. Laryngeal dysfunction may carely occur as part of the picture of acute dystonia and, if so, usually with dyspnoea. We describe a case of acute dystonia with atypical onset without relation to changes in dosage and with laryngeal involvement with aphonia, but without dyspnoea.     

Interferon alpha-2b in the treatment of chronic hepatitis C: early experience

The antiviral and immunomodulatory effects of interferon were assessed in the treatment of chronic Hepatitis C in multi-ethnic patients to prevent viral replication and chronic liver damage. Three million units of recombinant interferon alpha-2b were administered three times a week for 48 weeks to a group of 9 active Hepatitis C patients. A clinical response was defined as normalization of serum ALT values. Serum was frozen and stored for Hepatitis C viral assays. Four patients normalized their liver functions. When viral levels were measured only two patients had unmeasurable levels of HCV RNA after treatment. Therapeutic results were observed and much work needs to be done to improve therapy because a serious epidemic is predicted for the future.     

Nitric oxide is present in exhaled breath in humans: direct GC-MS confirmation

The physiological role of nitric oxide (NO) is being investigated in many experimental and clinical settings. There is considerable evidence that NO is involved in the regulation of lung vascular function. In addition there are many studies reporting the beneficial effect of NO inhalation. NO formed from L-Arginine has been detected in exhaled breath using indirect mass spectrometry and chemiluminescence. Both methods provided good evidence for the presence of NO in breath samples but were not unequivocal. We therefore developed a method using gas-chromatography-mass spectrometry which allowed us to measure trace levels of NO in air and breath. Eight healthy volunteers supplied numerous breath samples for analysis. A clear peak for nitric oxide was observed in seven volunteers. The mean level was 13ppb (n = 7, range < 2 to 19ppb). This data is in good agreement with our previous data and unequivocally confirms the presence of nitric oxide in human breath.     

A comparative evaluation of halo pin designs in an immature skull model

To design an improved halo pin for use in pediatric patients, three commonly used halo pins were evaluated with a mechanical testing apparatus and segments of prepared fetal calf skull. The pins were driven through the bone segments while the load at the bone-pin interface was measured. New pins were designed with respect to pin tip and flange width and similarly compared. Mean maximum loads to penetration, normalized for bone segment thickness, were 55.6 N/mm for the PMT Corporation pin, 61.5 N/mm for the Bremer pin, and 73.6 N/mm for the Ace pin. Four new, short tipped pins were designed and compared with the Ace pin, and there was no significant difference. Finally, four new pins were designed with varying flange widths. Mean maximum loads, normalized for bone segment thickness, were 68.9 N/mm for the 4.2 mm flange, 72.2 N/mm for the 4.7 mm flange, 92.9 N/mm for the 5.2 mm flange, and 96.4 N/mm for the 5.7 mm flange. The findings of this investigation are clinically important because they may help to explain the variability in the complication rates seen with the use of different halo systems in children. The three halo pins currently on the market have different pin designs, including tip lengths and flange distances, which contribute to the difference in load to penetration for each pin. The new, wide flanged, short tipped halo pin design might decrease the complication rate of halo use in children by providing an improved capacity to resist penetration despite increased loads of application.     

Effects of estrogen on tight junctional resistance in cultured human umbilical vein endothelial cells

OBJECTIVE: To study the effects of estrogen on transendothelial paracellular permeability in women. METHODS: Human umbilical vein endothelial cells (HUVEC) obtained from women were grown on filters. The paracellular permeability characteristics were determined in terms of changes in the permeability to the polar acid pyranine (Ppyr) and as changes in the transendothelial electrical resistance (RTE). Tight junctional resistance characteristics were assayed by lowering luminal NaCl and measuring the dilution potential, and were expressed as the ratio of monoion mobility uCl/uNa (cation selectivity). RESULTS: Low extracellular calcium and hyperosmolarity increased Ppyr and decreased RTE. The former but not the latter condition abolished the endothelium-specific cation selectivity. Treatment with 10 nM of estradiol-17 beta had no effect on RTE, but it increased the cation selectivity. The effect of estradiol required 1-6 hours' incubation with the hormone; it was dose dependent and saturable, with a median effective concentration of estradiol of 1 nM. Diethylstilbestrol, but not estriol, could mimic the effect of estradiol, and the estrogen receptor antagonist ICI-182, 780 blocked it. CONCLUSION: Cultured HUVEC cells form patent tight junctions. Estrogens increase the cation selectivity across HUVEC cultures. The effect of estrogen may be mediated by an estrogen receptor. These effects may be important for vasculoprotection in cases of sudden changes in ions levels across the capillary wall, such as ischemia or reperfusion.