Terminal restriction fragments of telomere are detectable in plasma and their length correlates with clinical status of ovarian cancer patients.

Free plasma DNA was extracted and terminal restriction fragment (TRF) sequences were amplified by polymerase chain reaction in 32 patients with stage 3 and stage 4 ovarian cancer and 45 healthy controls. Three peaks of TRF were identified and the size of the largest peak (peak 1) correlated with cancer telomere length in cancer patients. Average size of peak 1 in cancer patients was significantly shorter than in controls. When plasma TRF peak 1 was tested pre- and post-treatment, the average pre-treatment size was 8.7 +/- 0.5 Kb, lengthening significantly post-treatment. In long-term survivors of ovarian cancer (10-year disease-free survival), plasma TRF length was the same as in normal controls. The results suggest that the plasma TRF in ovarian cancer patients is of tumour origin. Its determination may reflect tumour cell viability in the host.

The nonhelical tail domain of keratin 14 promotes filament bundling and enhances the mechanical properties of keratin intermediate filaments in vitro.

Keratin filaments arise from the copolymerization of type I and II sequences, and form a pancytoplasmic network that provides vital mechanical support to epithelial cells. Keratins 5 and 14 are expressed as a pair in basal cells of stratified epithelia, where they occur as bundled arrays of filaments. In vitro, bundles of K5-K14 filaments can be induced in the absence of cross-linkers, and exhibit enhanced resistance to mechanical strain. This property is not exhibited by copolymers of K5 and tailless K14, in which the nonhelical tail domain has been removed, or copolymers of K5 and K19, a type I keratin featuring a short tail domain. The purified K14 tail domain binds keratin filaments in vitro with specificity (kD approximately 2 microM). When transiently expressed in cultured cells, the K14 tail domain associates with endogenous keratin filaments. Utilization of the K14 tail domain as a bait in a yeast two-hybrid screen pulls out type I keratin sequences from a skin cDNA library. These data suggest that the tail domain of K14 contributes to the ability of K5-K14 filaments to self-organize into large bundles showing enhanced mechanical resilience in vitro.

Follicle-stimulating hormone promotes the growth of human epithelial ovarian cancer cells through the protein kinase C-mediated system.

We have previously described that follicle-stimulating hormone (FSH) stimulated the growth of human epithelial ovarian cancer tissues and cells. In order to determine the signaling pathway on FSH action in ovarian cancer, we used an epithelial ovarian cancer cell line (HRA line) which constitutively FSH receptors (FSHRs). FSH significantly increased cell proliferation (230.1 +/- 20.5%, P < 0.05) and (3)H-thymidine uptake (443.5 +/- 35.1%, P < 0.01). 1-(5-Isoquinolinesulfonyl)-2-methyipiperazine (H7, 1 5 nM), staurosponine (STR, 5 nM) and calphostin C (5 nM), specific protein kinase C (PKC) inhibitors, significantly suppressed the FSH-stimulated cell growth (120.2-140.2%, P < 0.05) and (3)H-thymidine uptake (140.5-173.9%, P < 0.05), whereas N-(2-guanidinoethyl)-5-isoquinoline-sulfon-amide (HA1004, l5 nM), which is a derivant of H7 and inhibits most of protein kinases except PKC, showed no effect on the FSH-stimulated cell growth and (3)H-thymidine uptake. A pretreatment with 12-0-tetradecanoylphorbol-13 acetate (TPA, 100 ng/ml) or STR (20 nM) significantly suppressed the subsequent FSH-stimulated cell growth (TPA; 152.3 +/-10.3%, STR; 160.4 +/- 15.9%, P < 0.05) and (3)H-thymidine uptake (TPA; 250.4 +/-18.3%, STR; 208.7 +/- 15.9%, P < 0.05). STR abolished the suppression of TPA preincubation on the subsequent FSH-stimulated cell growth and (3)H-thymidine uptake. HRA cells constitutively expressed PKCalpha but not PKCbeta nor PKCgamma. The levels of either expression of PKCalpha protein and mRNA were significantly amplified by FSH. These data suggest that stimulation of PKCalpha transcription is involved in the FSH-stimulated cell growth and DNA synthesis in epithelial ovarian cancer cells.

[Inhibition of corneal neovascularization--the possibility of inhibition by drug administration].

PURPOSE: The anti-angiogenic activity of FR 118487, a new synthetic analog of Scolecobasidium arenarium products, was examined in Japanese white rabbit cornea. METHODS: We studied both systemic and locally administered FR 118487 (ointment) in a keratoplasty model consisting of corneal neovascularization after implantation of a Wister rat cornea into a rabbit cornea. RESULTS: Two weeks after the implantation, the maximum length of neovascularization was 3.4 +/- 0.3 mm in control corneas, 0.1 +/- 0.0 mm with systemic FR 118487 administration (10 mg/day) (p < 0.01), 0.1 +/- 0.1 mm with 10% FR118487 ointment (p < 0.001), 1.0 +/- 0.2 mm with 3% FR 118487 ointment (p < 0.001), and 0.9 +/- 0.9 mm with 1% FR 118487 ointment (p < 0.02). CONCLUSION: FR 118487 had a significant effect on inhibition of corneal neovascularization.

[Inhibition of corneal neovascularization by a new analog of fungal product].

The anti-angiogenic activity if FR 118487, a new synthetic analogue of Scolecobasidium arenarium products, was examined in corneas of Japanese white rabbits. We studied locally administered FR 118487 with hydron pellets in two models; corneal neovascularization after implantation of a CuCl2 pellet (CuCl2-induced model) and a Wistar rat's cornea (keratoplasty model) into a rabbit cornea. In the CuCl2-induced model, maximum length of neovascularization was 0.08 +/- 0.10 (mean +/- standard deviation) mm with FR 118487 (control 2.84 +/- 0.13 mm) at 1 week after the implantation. In the keratoplasty model, maximum length of neovascularization was 0.05 +/- 0.05 mm with FR 118487 (control 3.33 +/- 0.18 mm) at 2 weeks after the implantation. In both models, FR 118487 had a significant (p < 0.01) effect on inhibition of corneal neovascularization.

[The presence of IgE on limbal Langerhans cells in patients with atopicdermatitis].

Limbal conjunctival biopsies from 8 patients with atopic dermatitis and from 5 age-matched healthy individuals undergoing cataract or retinal detachment surgery were analyzed by light microscopy and immunological techniques. They were immuno-double labelled with anti-CD1a and anti-IgE or anti-CD23 (IgE receptor). In the specimens from atopic dermatitis 24 approximately 75% of positive anti-CD1a staining cells were double-stained by anti-IgE. Weak positive immuno-double stained cells with anti-CD23 were also observed, but less than with anti-IgE. The ratio of positive anti-IgE double-stained cells to positive anti-CD1a stained cells seemed to be parallel to serum IgE level, but not significant. The presence of IgE and CD23 (IgE receptor) on conjunctival Langerhans cells seems to have a positive effect on IgE-dependent antigen presentation.

[Experimental study on the effects of endotoxemia as a retardation-factor influencing on the decrease of serum bilirubin after the relief of obstructive jaundice].

UNLABELLED: I examined the effects of endotoxemia influencing on obstructive jaundice and the decrease of serum bilirubin after the relief of it. Experimental obstructive jaundice and its alleviation by external biliary drainage was made in Donryu-rats. Serum bilirubin was higher (p < 0.01) in the group treated by continuous infusion of low-dose endotoxin (LPS E. coli 0111:B4, 6 micrograms/hr/100gBW) during 72 hours biliary obstruction (10.48 +/- 1.54mg/dl) than in the control group (6.76 +/- 0.71mg/dl). After the relief of biliary obstruction, 4 kinds of experimental conditions were set up as follows: CONTROL; infusion of sterile saline, ET; continuous infusion of low-dose endotoxin, ET + UDCA; continuous infusion of endotoxin and ursodeoxycholic acid (UDCA, 10mg/hr/100gBW) through another intravenous tube simultaneously, ET + MP; continuous infusion of endotoxin after one-shot injection of methylprednisolone (MP, 3mg/100gBW). Serum bilirubin 7.5 hours after the relief of biliary obstruction was as follows: CONTROL: 0.48 +/- 0.18mg/dl, ET: 3.41 +/- 1.13mg/dl, ET + UDCA: 2.80 +/- 1.28mg/dl, ET + MP: 1.18 +/- 0.50mg/dl. ET-group showed retardation of the decrease of serum bilirubin (p < 0.01). MP showed improvement of the impaired decreasing rate of serum bilirubin by endotoxemia (p < 0.01). Bile-output from the external biliary drainage after the relief of biliary obstruction was decreased significantly in the ET-group. ET + UDCA-group showed remarkable increase of the bile-output, but no increase of excretion of bilirubin in the bile compared with ET-group. While ET + MP-group showed improvement of the bile-output and increase of excretion of bilirubin in the bile compared with ET-group.(ABSTRACT TRUNCATED AT 250 WORDS)

[A research on the cholestasis caused by continuous endotoxemia].

Intrahepatic cholestasis is often observed in patients without obstruction of the bile duct, who are suffering from severe prolonged infection in the field of peptic surgery. Clinical data were analyzed in recently experienced 18 cases which showed this kind of jaundice. In those case, high rates of endotoxemia and high rates of gram negative bacilli were seen. This fact made us infer that endotoxins might cause jaundice. In order to clarify the mechanism of the jaundice, we made an experimental model of persistent endotoxemia in rats. Low-dose endotoxin was infused continuously to Donryu-rats and bile-output was observed with external bile-guiding tube for 24 hours. In the endotoxin group, bile-output was significantly decreased whereas it was not changed in the control group. In addition, serum bilirubin was elevated in the endotoxin group, whereas it did not change in the control group. Blood-flow of liver tissue and systemic arterial blood pressure did not show any severe decrease under the continuous endotoxemia. Data of bile-output and bile acid showed bile acid independent flow might be depressed by endotoxin infusion. This model was thought to be under non-shock condition and useful to investigate jaundice seen in patients under continuous endotoxemia.

Subcultured astrocytes suppress the proliferation of neuroblasts in vitro.

The effects of subcultured astrocytes on the proliferation of neuronal precursor cells (neuroblasts) from rat embryonic cerebral hemispheres were examined. The survivability of neurons and the neurite outgrowth were significantly enhanced by the subcultured astrocytes compared to those of neurons plated on poly-L-lysine-coated coverslips. The incorporation of [3H]thymidine into neuroblasts was remarkably suppressed by the subcultured astrocytes indicating that the astrocytes inhibit the proliferation of neuroblasts. These results suggest that astrocytes enhance the maturation of neuroblasts possibly via either cell-cell contact or trophic substances.

Septal deafferentation enhances the neurotrophic effects of rat hippocampus on cultured neural cells from the central nervous system.

Neurotrophic effects (NTEs) of various brain regions of 4-week-old rats were examined in primary culture of rat embryonic cerebral hemispheres. Extracts of the hippocampus, brainstem and septal nucleus highly enhanced the survivability of neuronal cells and the division of non-neuronal cells by 9 days. The septohippocampal tract (fimbria fornix) was cut and the effect on the neurotrophic activity in the hippocampus was examined. The NTEs of hippocampal extracts remained unchanged 3 days after septal deafferentation, was significantly increased by 7 days, peaked at 14 days and returned to the basal level by 21 days.