Tumor necrosis factor-alpha inhibits human chorionic gonadotropin secretion.

The placenta is a major source of tumor necrosis factor-alpha (TNF alpha) and is rich in TNF alpha receptors. TNF alpha inhibited hCG secretion by normal chorionic villi at 6 weeks gestation, but chorionic villi contain the heterogenous cell population. To investigate the direct effects of TNF alpha on trophoblasts, the NUC1 choriocarcinoma cell line was used as an in vitro placental cell model. TNF alpha also inhibited hCG secretion by NUC1 cells at concentrations from 1-100 U/mL. TNF alpha at concentrations of 1, 10, and 100 U/mL significantly decreased hCG secretion for 24 h to 88%, 81%, and 71% of the control level, respectively. The expression of hCG beta mRNA was also decreased to 23% of the control level after 24 h of TNF alpha treatment (100 U/mL). Inhibition occurred after 12 h of TNF alpha treatment (100 U/mL). Two measurements, trypan blue dye exclusion and 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide reduction, revealed that these inhibitory effects were not due to the cytotoxic activity of TNF alpha on NUC1. In conclusion, TNF alpha reduces hCG secretion in vitro.

Expression of HLA class I and beta 2-microglobulin on human choriocarcinoma cell lines: induction of HLA class I by interferon-gamma.

The trophoblast cells which contact with maternal tissue express HLA class I either weakly or not at all. This phenomenon is beneficial for the protection of the fetus from immunological rejection by the mother. However, the mechanisms of reduced expression of HLA class I are unexplained. Therefore, we utilized choriocarcinoma cells to analyse the suppressive mechanisms to HLA class I expression in trophoblasts. In this paper we reported the heterogeneity of HLA class I expression in five choriocarcinoma cell lines. The levels of surface HLA class I molecules correlate approximately with mRNA of HLA class I heavy chain, and all choriocarcinoma cells produce moderate amounts of surface and secreted beta 2 microglobulin. It was also found that interferon gamma upregulates HLA class I molecules and mRNA in two choriocarcinoma cell lines, NUC1 and HCCM5.

Comparison of Biobrane and Scarlet Red dressings for treatment of donor site wounds.

Two methods of autograft donor site management were tested in 31 patients with burn injuries. Two donor sites of equivalent size were respectively covered with Biobrane and Scarlet Red, two commercial dressing materials available for donor wound coverage. Pain, the amount and type of exudate developing underneath the dressing, adherence, and cellulitis were evaluated on a daily basis, and healing time was determined. Biobrane proved superior to Scarlet Red with respect to control of pain, accumulation of exudate, and healing time; the materials were equal with regard to adherence to the wound, the character of exudate, and the appearance of cellulitis.

T-686, a novel inhibitor of plasminogen activator inhibitor-1, inhibits thrombosis without impairment of hemostasis in rats.

The aim of this study was to evaluate the antithrombotic potential of T-686 ((3E,4E)-3-benzylidene-4-(3,4,5-trimethoxy-benzylidene)-pyrr olidine-2,5-dione), a novel inhibitor of plasminogen activator inhibitor-1 (PAI-1), in rat thrombosis models. T-686 (0.1-100 mg/kg per day, p.o.) dose dependently decreased the weight of venous thrombi induced by a combination of stasis and hypercoagulability. The antithrombotic effect was enhanced by repeated administration of T-686. Warfarin (0.1 mg/kg per day for 3 days) also prevented thrombus formation. The antithrombotic action by warfarin was accompanied by prolongation of coagulation time, while no effect on coagulation time was observed in T-686-treated rats. T-686 lowered the activity of PAI-1 in plasma. In the arterio-venous shunt model, pretreatment with T-686 (10 mg/kg per day) or ticlopidine (100 mg/kg per day) for 8 days inhibited thrombus formation by 33% and 44%, respectively. T-686 had no effect on collagen-induced platelet aggregation ex vivo, while ticlopidine inhibited platelet aggregation. T-686 did not affect bleeding time at 10-100 times the antithrombotic dose, while warfarin dose dependently prolonged bleeding time at and around the antithrombotic dose. These results suggest that T-686 prevents thrombus formation in rats without impairment of hemostasis.

Preparation of oligozoospermic and/or asthenozoospermic semen for intrauterine insemination using the SpermPrep semen filtration column.

OBJECTIVE: To improve the quality of oligozoospermic and/or asthenozoospermic semen by the SpermPrep (Fertility Technologies Inc., Natick, MA) semen filtration column. DESIGN: The SpermPrep column was applied for semen manipulation in oligozoospermia and/or asthenozoospermia (sperm count less than 20 x 10(6)/mL, sperm motility less than 40%). After concentration of motile sperm using a 40% Percoll density gradient centrifugation, the sperm suspension was filtered through the SpermPrep column. The percentage yield of motile sperm by the SpermPrep method was compared with those by a two-layer Percoll density gradient (Pharmacia, Uppsala, Sweden) centrifugation and a swim-up method. Infertile couples with poor quality semen were treated with intrauterine insemination (IUI) with motile sperm by the three preparations through three cycles. SETTING: Department of Obstetrics and Gynecology, Osaka University Hospital. PATIENTS: Twenty-one couples with long-standing infertility because of poor quality semen. MAIN OUTCOME MEASURE: Recovery of motile sperm, sperm motility, and outcome of IUI were evaluated among three semen preparations. RESULTS: Motility was improved by the SpermPrep method in 32 of 33 cases of oligozoospermia and/or asthenozoospermia. Percentage yield of motile sperm by the SpermPrep method was significantly greater than those by the two-layer Percoll density gradient and swim-up methods (42.7 +/- 4.6 versus 22.1 +/- 3.1 and 13.8 +/- 3.5), but there is no significant difference in the sperm motility among three semen preparations. After one treatment cycle for each preparation, 2 of 21 women conceived after IUI with motile sperm separated in the SpermPrep method. CONCLUSIONS: The SpermPrep method is an improved semen manipulation method for oligozoospermia and/or asthenozoospermia.

Selection of acrosome-reacted sperm with MH61-immunobeads.

Separation of acrosome-reacted viable sperm is an important problem in the analysis of sperm function and the micromanipulation of gametes such as the microinjection of sperm into ooplasm and subzonal sperm insertion. In the present study, we collected acrosome-reacted sperm selectively using paramagnetic immunobeads, referred to as MH61-beads, which attach themselves to the head of the acrosome-reacted sperm and are recovered by a magnet. The maximum number of sperm was recovered by the MH61-beads when 3 x 10(5) sperm/100 microliters and 6 x 10(5) MH61-beads were coincubated for 1 hour. The recovery of sperm was significantly correlated with the percentage of the acrosome reaction in the capacitation medium. After incubation for 12 hours, 2.4-9.0 x 10(4) sperm could be collected, and the percentage of viable and acrosome-reacted sperm was 28-60%. To improve the recovery of the acrosome-reacted sperm, we utilized follicular fluid, which induces the acrosome reaction. Addition of follicular fluid to the capacitation medium at a final concentration of 20% induced the acrosome reaction and increased the recovery of acrosome-reacted sperm significantly. In conclusion, separation of acrosome-reacted sperm by the MH61-bead-binding method could become a useful technique in research of sperm function and gamete micromanipulation.

Regulation of classical HLA class I genes in human choriocarcinoma cells by nuclear proteins binding to MHC class I regulatory elements.

PROBLEM: The regulation of classical HLA class I genes in choriocarcinoma have been reported. METHODS: We determined whether four choriocarcinoma cell lines expressed classical HLA class I or HLA-G by a reverse transcription-polymerase chain reaction (RT-PCR) and studied the regulatory mechanism of classical class I using a gel mobility shift assay. RESULTS: NUC1 and SCH expressed classical class I but not HLA-G. GCH1 and Jar did neither. Nuclear protein binding to the class I regulatory element (CRE) was detected in NUC1 and SCH. Interferon-gamma augmented both classical class I expression and the DNA-protein complex in NUC1. The DNA-protein complex was not observed in GCH1, and Jar showed a CRE-binding protein with different electrophoretic mobility and binding affinity from that of SCH and NUC1. CONCLUSION: The CRE is one of the regulatory elements of classical HLA class I genes in choriocarcinoma cells.

HLA expression on human ejaculated sperm.

The expression of human leukocyte antigen (HLA) on human ejaculated sperm has been investigated by using cellular binding radioimmunoassay (CB-RIA) with sperm samples purified by Percoll discontinuous gradient and monoclonal antibodies specific to the frame work of HLA molecules. The results in packed sperm obtained from ten healthy males indicated that HLA class II and its subclasses, DP, DQ, and DR, were expressed at a low level on human ejaculated sperm, while HLA class I was not detected. Furthermore, we could not detect the individual variation of HLA expression on ejaculated sperm in the cases of twenty healthy males. K562, which is B lymphoid cell line and lacks HLA class I molecules, showed detectable amounts of HLA class I molecules after exposure to interferon gamma (IFN-gamma) and/or tumor necrosis factor alpha (TNF-alpha). However, the treatment of sperm with IFN-gamma and/or TNF-alpha had no effect on the expression of HLA class I and class II molecules.

Clinical experience with Biobrane biosynthetic dressing in the treatment of partial thickness burns.

Biobrane, a synthetic, bicomposite wound dressing, has been used to treat 17 patients with partial thickness burn wounds covering 0.5-12.5 per cent of the total body surface area (mean 4.4 per cent). In 16 patients we found complete healing of the wound after removal of the dressing, 6-15 days after the injury. In one patient, the wounds were determined on the third day post-injury to be deeper than initially suspected, and she was taken to the operating room for surgical debridement and grafting. Biobrane is an effective wound covering for clean, superficial partial thickness burns of limited extent; the simultaneous use of topical antimicrobial agents for such wounds is not necessary.