Identification of extracellular matrix components and their integrin receptors in the human fetal adrenal gland.

The development of the human fetal adrenal gland is characterized by a gradient of mitotic activity, cell migration, and cell apoptosis, all of which dictate its particular function. Such plasticity may possibly be under the control of the extracellular environment. The goal of this study was to identify components of the extracellular matrix in second-trimester fetal adrenal glands. Whereas collagen IV was expressed evenly throughout the gland, both fibronectin and laminin demonstrated a mirror-imaged distribution, with higher expression of fibronectin in the central portion and laminin at the periphery of the gland. The integrin subunit alpha1 was found mainly in the definitive zone and the alpha2-subunit mainly in the transitional zone, whereas integrin alpha3 (which binds both fibronectin and laminin) was detected only in the fetal zone. The beta2-subunit was observed solely in chromaffin cells. Such specific gradients of integrin and MEC component expression suggest that the extracellular environment does play a definite role during adrenal gland development. Indeed, compared with that in untreated plastic dishes, ACTH stimulation of dehydroepiandrosterone sulfate and cortisol was enhanced by collagen IV. In addition, fibronectin enhanced dehydroepiandrosterone sulfate but decreased cortisol secretion, compared with collagen IV substrates. These results provide fundamental insight into the contribution of the microenvironment in cellular processes leading to fetal adrenal gland development.

Dimethyl sulfoxide affects the selection of splice sites.

Depending on the cell lines and cell types, dimethyl sulfoxide (Me2SO) can induce or block cell differentiation and apoptosis. Although Me2SO treatment alters many levels of gene expression, the molecular processes that are directly affected by Me2SO have not been clearly identified. Here, we report that Me2SO affects splice site selection on model pre-mRNAs incubated in a nuclear extract prepared from HeLa cells. A shift toward the proximal pair of splice sites was observed on pre-mRNAs carrying competing 5'-splice sites or competing 3'-splice sites. Because the activity of recombinant hnRNP A1 protein was similar when added to extracts containing or lacking Me2SO, the activity of endogenous A1 proteins is probably not affected by Me2SO. Notably, in a manner reminiscent of SR proteins, Me2SO activated splicing in a HeLa S100 extract. Moreover, the activity of recombinant SR proteins in splice site selection in vitro was improved by Me2SO. Polar solvents like DMF and formamide similarly modulated splice site selection in vitro but formamide did not activate a HeLa S100 extract. We propose that Me2SO improves ionic interactions between splicing factors that contain RS-domains. The direct impact of Me2SO on alternative splicing may explain, at least in part, the different and sometimes opposite effects of Me2SO on cell differentiation and apoptosis.

Enhancement of the biodegradability of model wastewater containing recalcitrant or inhibitory chemical compounds by photocatalytic pre-oxidation.

m-Dinitrobenzene, diphenylamine and resorcinol, three aromatic compounds found inhibitory or recalcitrant to biological treatments, were chosen as model chemicals for this study on the integration of photocatalytic-biological treatments. The degradation of each of these compounds was achieved by ultraviolet photocatalytic oxidation, leading to the formation of intermediate compounds. The photocatalytic treatment was performed in a TiO2 slurry reactor containing an aqueous solution of one of the three chemicals. The biodegradability of model wastewater treated photocatalytically was measured in terms of BOD1/TOC. Intermediate compounds that appeared at early stages of the photocatalytic degradation of m-dinitrobenzene or diphenylamine seemed to be more inhibitory than the parent compounds but this was not the case for resorcinol. A substantial improvement in BOD1/TOC could be achieved, but it required the mineralization of at least 80% of the organic carbon originally in the water. Microtox toxicity results confirmed the BOD1/TOC trends for diphenylamine.

Preliminary clinical experience with a thermal balloon endometrial ablation method to treat menorrhagia.

OBJECTIVE: To evaluate the clinical effectiveness and safety of a thermal balloon system to ablate the endometrium. METHODS: All 18 patients were candidates for hysteroscopic endometrial ablation or hysterectomy for menorrhagia and consented to a trial of the balloon technique of ablation. All procedures were done in the operating room under general anesthesia, except in one patient who had regional and another who had local anesthesia with analgesia. Follow-up of 6-34 months is reported. RESULTS: Fifteen subjects (83%) reported significant reduction in bleeding or amenorrhea. Two patients underwent subsequent hysterectomy and one a follow-up hysteroscopic examination with biopsy. Histology in these three cases showed areas of scar as well as areas of normal endometrial histology. In one uterus, the entire cavity and the endometrium were normal. The others had endometrial bands of scar and some contraction of the cavity. CONCLUSIONS: Based on follow-up results, the frequency of successful reduction of bleeding and/or amenorrhea in this small series is comparable to hysteroscopic methods of endometrial ablation. There were no complications. A larger trial is warranted to compare this method to hysteroscopic endometrial ablation.

The endometrial ablator: a new instrument.

A thermal balloon and control system designed to produce endometrial ablation blindly is described. A latex balloon on a plastic catheter is inserted into the uterus and connected to a control unit. The unit monitors the pressure and temperature of 5% dextrose in water, which has been injected into the balloon to make it conform to the size and shape of the endometrial cavity. The balloon contains a shielded heating element that is activated to heat the liquid in the balloon to a temperature of 92C. The pressure control deactivates the heating element if the pressure falls below 45 mmHg or rises above 165 mmHg. A timer controls and measures the elapsed interval of heating. The device was tested in human uterine specimens for the potential for uterine perforation, uterine rupture, and thermal effects. Subsequently, the device was tested in six patients in Mexico and four patients in London during hysterectomy just after the abdomen was opened. Thermistor probes were placed at various loci in the uterus to monitor temperature during activation of the thermal balloon. Serosal temperatures were unchanged and endometrial temperatures rose to about 90C. The extent of uterine tissue damage was determined in Mexico City by the zone of visible coagulation of the cut wall of the uterus following removal. In London, tissue diaphorase was measured to determine the depth of destruction of the cellular oxidative enzymes. These measurements varied from 3.3-10 mm under the conditions of time and temperature used. The safety features and the potential for clinical application are discussed.

Effect of inhibition of hepatic fatty acid oxidation on metabolic and hormonal responses to exercise in rats.

The purpose of the present study was to assess the effects of 2-mercaptoacetate (MA), an inhibitor of hepatic fatty acid oxidation, on the metabolic and pancreatic hormone response to a prolonged (3 h) swimming exercise. All rats were first adrenodemedullated and were either submitted for 3 weeks to a normal (5% fat) or a medium-fat diet (MF; 18% fat). After being submitted to an exercise swimming habituation programme for one week, rats under both dietary conditions were either injected with a bolus dose of MA (600 mumol/kg; ip) or with a saline solution. MA and saline injected rats were either sacrificed after a 3-h swimming exercise or after a 3-h resting period. Administration of MA was associated with a lower level of beta-hydroxybutyrate after exercise in rats fed the MF diet, higher resting and exercising blood glucose levels in rats fed the MF diet, and higher resting and exercising levels of hepatic glycogen in rats fed a normal diet. There were, however, no significant effects of MA on free fatty acid, insulin, glucagon, epinephrine, and norepinephrine concentrations in both dietary conditions either at rest or after exercise. Therefore, the present data do not provide any evidence that the administration of MA, an inhibitor of hepatic fatty acid oxidation, influences the pancreatic hormonal response to exercise. There was also no evidence of a lowering effect of MA on blood glucose levels during exercise.

Trials with the FEMCEPT method of female sterilization and experience with radiopaque methylcyanoacrylate.

A previous report described the development of a blind method to deliver methylcyanoacrylate (MCA) transcervically. Using 0.6 ml of a stable MCA whose polymerization time was closely controlled, we reported a 78% bilateral tubal closure rate in 23 cases with hysterosalpingographic control. Subsequent to the previous report, we initiated a study in which patients were randomly assigned to one of three treatment groups: a single MCA injection, a single MCA injection after uterine lavage, or two MCA injections 1 month apart. In addition, a radiopaque MCA has been developed with which it is possible to determine tubal entry after its application by means of the FEMCEPT device. Patients treated with radiopaque MCA have been studied to determine whether it is possible to predict tubal closure on the basis of tubal entry and distribution patterns. The results of these studies and their implications for contraceptive effectiveness of the FEMCEPT/MCA system will be reported.

[An outpatient approach to female sterilization using methylcyanoacrylate (author's transl)]. / Sterilization feminine par methylcyanoacrylate, en ambulatoire.

PIP: The article reports on the results of 131 sterilizations by tubal occlusion obtained by injection of MAC (methylcyanoacrylate). The procedure is conducted by using a special device, called a Femcept. In the series presented here there were no complications and no side effects, and the obstruction of the tubes, tested by hysterosalpingography, was bilateral in 78% of cases. A second treatment with MAC can be done. This method does not involve surgery, anesthesia, or any specific experence or equipment; it is safe, quick, economic, and can be done on an outpatient basis.^ieng

An outpatient approach to female sterilization with methylcyanoacrylate.

MCA is a tissue adhesive which can be delivered transcervically to the Fallopian tubes by means of the FEMCEPT device. In the patients treated with this system, both prior to hysterectomy and on an ambulatory basis, there have been no significant complications or side effects. In the most recent series of ambulatory patients treated with the REMCEPT-MCA system, the bilateral tubal closure rate was 78%.

PIP: Clinical trials of methylcyanoacrylate (MCA), a tissue adhesive injected to occlude fallopian tubes for tubal sterilization, were conducted in ambulatory patients and are presented. 3 small series of patients are described; most received .4-ml injections, but 23 of the 131 total subjects received .6-ml injections. Outcome was determined by hysterosalpingography. Bilateral closure rate after 1 treatment in series 1 (n=28) was 64%. In the 2nd series the bilateral closure rate was 54% after 1 injection; after a 2nd injection in the 2nd series, the rate was 78%, but overall the closure rate for this group after 2 injections was 65% (n=102). In the 3rd series, bilateral closure rate was 78% after 1 injection; this series used .6-ml injections and represents a dramatic improvement over the preceding series. Complications include pain at site of injection, mild to moderate cramping, and transient vagal reaction. The procedure takes 5 minutes or less. 2 uterine pregnancies have occurred to date, and no ectopic pregnancies were reported. MCA is a low-viscosity fluid in a monomeric form which rapidly polymerizes when placed in contact with body fluids.

Single-application fertility-regulating device: description of a new instrument.

An instrument is described with which it is possible to deliver low-viscosity material consistently to the interstitial and isthmic portions of the Fallopian tube in a blind fashion on an outpatient basis. Considerable care with instrument design has been taken in order to reduce potential complications and to ensure precision. The efficiency of this system in delivering a variety of dyes has been demonstrated in hysterectomy specimens with the use of laparoscopic control. We believe this system is simple to use and may be found acceptable for application by relatively unskilled personnel. If it produces a high degree of tubal closure without significant complications or pregnancies, it should be an important addition to the techniques for limiting family size and population growth.