Identification of a novel interleukin-6 response element containing an Ets-binding site and a CRE-like site in the junB promoter.

Interleukin-6 (IL-6) activation of the immediate-early gene junB has been shown to require both a tyrosine kinase and an unknown 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H7)-sensitive pathway. Here we report the identification and characterization of an IL-6 immediate-early response element in the junB promoter (designated JRE-IL6) in HepG2 cells. The JRE-IL6 element, located at -149 to -124, contains two DNA motifs, an Ets-binding site (EBS) (CAGGAAGC) and a CRE-like site (TGACGCGA). Functional studies using variously mutated JRE-IL6 elements showed that both motifs were necessary and sufficient for IL-6 response of the promoter. The EBS of the JRE-IL6 element (JEBS) appears to bind a protein in the Ets family or a related protein which could also form a major complex with the EBSs of the murine sarcoma virus long terminal repeat or human T-cell leukemia virus type 1 long terminal repeat. The CRE-like site appears to weakly bind multiple CREB-ATF family proteins. Despite the similarity in the structure between the JRE-IL6 element and the polyomavirus enhancer PyPEA3, composed of an EBS and an AP1-binding site and known to be activated by a variety of oncogene signals, JRE-IL6 could not be activated by activated Ha-Ras, Raf-1, or 12-O-tetradecanoylphorbol-13-acetate. We show that IL-6 activates JRE-IL6 through an H7-sensitive pathway that does not involve protein kinase C, cyclic AMP-dependent kinase, Ca(2+)- or calmodulin-dependent kinases, Ras, Raf-1, or NF-IL6 (C/EBP beta). The combination of JEBS and the CRE-like site appears to form the basis for the selective and efficient response of JRE-IL6 to IL-6 signals, but not to signals generated by activated Ha-Ras, Raf-1, or protein kinase C.

Complete epiretinal membrane separation in highly myopic eyes with retinal detachment resulting from a macular hole.

PURPOSE: Epiretinal membranes (ERMs) in highly myopic eyes may result in macular holes and subsequent retinal detachment. However, removing friable, thin ERMs from detached retinas is often difficult. We report the efficacy of a diamond-dusted silicone cannula in the removal of ERMs from detached retinas. METHODS: Seventeen consecutive highly myopic eyes (16 patients) with retinal detachment underwent pars plana vitrectomy with gas tamponade. Peeling of the ERM adjacent to the macular hole was performed using either conventional tools (n = 11) or a diamond-dusted silicone cannula (n = 6). The rate of complete membrane peeling and the effect of membrane removal on the anatomic success rate were compared between groups. RESULTS: Retinal reattachment occurred in 13 (92.9%) of the 14 eyes in which the ERM was removed completely; redetachment occurred in the other three eyes, with further surgical interventions in two eyes. The reattachment rate was significantly higher (P = .005) when ERM removal was complete than when there was residual ERM. In the initial surgery, the ERM was successfully removed in all 6 eyes (100%) in the diamond-dusted silicone cannula group and in 5 of 11 eyes (45.5%) in the conventional group (P = .041); the reattachment rate was 100% in the diamond-dusted silicone cannula group and 45.5% in the conventional group (P = .005). When a second surgery was performed, the use of the diamond-dusted silicone cannula was also better than conventional tools for removing the residual ERMs, resulting in retinal reattachment. CONCLUSION: In highly myopic eyes with a macular hole and subsequent retinal detachment, complete ERM removal is closely related to successful retinal reattachment. The diamond-dusted silicone cannula appears to be more effective than conventional tools for removing ERM and may increase the anatomic success rate. Because of the limitations of a small series, a prospective, randomized trial is required to confirm the current beneficial results of using a diamond-dusted silicone cannula.

Migrastatin, a new inhibitor of tumor cell migration from Streptomyces sp. MK929-43F1. Taxonomy, fermentation, isolation and biological activities.

A new compound, migrastatin, was isolated from a cultured broth of Streptomyces sp. MK929-43F1, as an inhibitor of tumor cell migration. It was purified by column chromatographies on silica gel and Sephadex LH-20 and HPLC. Migrastatin has the molecular formula of C27H39NO7 consisting of 14-membered macrolide and glutarimide moiety. It inhibited spontaneous migration of human esophageal cancer EC17 cells. Migration inhibitory activity of migrastatin was not dependent on cytotoxicity or inhibition of protein synthesis.

Cytokine effects on phagocytosis of rod outer segments by retinal pigment epithelial cells of normal and dystrophic rats.

PURPOSE: Phagocytosis of rod outer segments (ROS) is an important function of retinal pigment epithelial (RPE) cells. Since the details of the process are not fully known, we studied effects of cytokines produced by RPE and photoreceptor cells on phagocytosis of ROS by rat RPE cells. METHODS: RPE cells were isolated and cultivated from two strains of rats: Sprague-Dawley (SD) rats with normal phagocytosis and Royal College of Surgeons (RCS) rats, which have genetic deficiencies in ROS phagocytosis. A double immunofluorescence staining technique was used to study the effects in vitro of several cytokines on phagocytosis of ROS. RESULTS: We found that transforming growth factor beta-1 (TGF-beta 1) had dose-dependent effects on RPE cells of both strains of rat: at a concentration of 10 ng/ml, TGF-beta 1 significantly (p < 0.01) reduced total ROS (to 74% of control in SD rats and to 51% of control in RCS rats), reduced bound ROS (to 56% of control in SD rats and to 48% in RCS rats), and increased the ratio of ingested ROS to total ROS (to 140% in SD rats but not significantly in RCS rats). Treatment of medium with anti-TGF-beta 1 antibody before incubation of RPE cells of SD rats with TGF-beta 1 decreased the magnitude of these effects. The cytokine acidic fibroblast growth factor (aFGF, 10 ng/ml) affected RPE cells of SD rats only, decreasing ROS ingested to 56% of control and the ratio of ingested ROS to total ROS to 64% of control. We also examined effects of basic fibroblast growth factor and insulin-like growth factor. None of the cytokines tested increased ingestion of ROS by RPE cells of RCS rats. CONCLUSIONS: Our results suggest that TGF-beta 1 and aFGF have roles in regulating ROS phagocytosis by normal and dystrophic RPE cells in the rat.

Inhibitory effect of acetic acid on growth of hyperthermophilic archaeon Pyrococcus furiosus.

The growth inhibition of Pyrococcus furiosus by acetic acid was stronger than that by hydrogen and could be described by a non-competitive inhibition model in which the inhibition constants of undissociated acetic acid, K(p) and n, were estimated to be 0.69 mM (25 mM total acetic acid at pH 6.5; pKa=4.96; 98 degrees C) and 1.0, respectively. In order to reduce the acetic acid inhibition, repeated-batch culturing was performed using a filtration module. This yielded 0.49 g of dry cells l(-1) after growth for 12 h after inoculation. It became impossible, however, to continue repeated-batch culturing manually because the time intervals for medium replacement became too short. In order to automatically maintain a low concentration of acetic acid, a perfusion culture was carried out in which medium feeding coupled to a pH-auxostat was performed. In this perfusion culture, it was possible to maintain the acetic acid concentration below 7.6 mM during exponential growth of P. furiosus, resulting in 1.8 g of dry cells l(-1) at 15 h after inoculation.

A study of familial occurrence of Behçet's disease with and without ocular lesions.

PURPOSE: To examine retrospectively the features of Behçet's disease patients with familial occurrence and make a comparison between familial Behçet's patients with ocular lesions and those without ocular lesions. METHODS: We sent questionnaires about Behçet's disease patients with familial occurrence to 114 hospitals that reported treating such patients in previous nationwide hospital surveys, and to 341 hospitals selected at random in Japan. RESULTS: We obtained reports on 83 Behçet's patients with familial occurrence. The positive rate of HLA-B51 was 53.1%. The positive rate of HLA-B51 among patients with ocular lesions was 64.0%, that in patients without ocular lesions was 14.3%, and that in patients with genital ulcers was 40.9%. The mean birth year and onset year of the patients with ocular symptoms was significantly lower than that of the patients without ocular symptoms. Although there was no significant difference between these two groups, parent-child involvement was more common among the patients without ocular lesions than in those with ocular lesions. CONCLUSIONS: The findings of this study suggest that the number of familial Behçet's disease patients with ocular lesions and high HLA-B51 positivity has been decreasing recently. To elucidate the etiology of familial occurrence, larger scale epidemiological studies and further molecular studies of Behçet's disease are needed.

The comparison of tuberculosis treatments: a short course therapy and the directly observed short course treatment (DOTS), East Java Province, Indonesia.

Tuberculosis has been given great attention as HIV/AIDS has increased. Because HIV causes a higher tuberculosis risk is becoming more and more important better tuberculosis treatment. This study aimed to compare the smear conversion rate between short course therapy and the tuberculosis treatment with directly observed short course therapy (DOTS), in East Java, Indonesia. The average smear conversion rate in short course therapy among 35,292 cases was 94.40% over 5 year period (1989/90-1993/94). The tuberculosis treatment with DOTS was started in 1994/95. In the first 2 years the smear conversion rate were 97.67% (42/43) and 98.00% (196/200), respectively. The smear conversion rate of the treatment with DOTS was significantly higher compared to a short course therapy (p-value: <0.001). Thus, tuberculosis treatment with DOTS should be promoted. The concept of supervision by health workers or health cadres should be applied considering mostly are given by family members. And there should be readiness of tuberculosis staff to do the treatment with DOTS in all levels to expand the coverage.

Factors associated with TT (tetanus toxoid) immunization among pregnant women, in Saparua, Maluku, Indonesia.

A cross sectional study was conducted at Saparua Health Center to determine factors associated with the administration of tetanus toxoid. In all, 64 pregnant women were recruited. The majority of the women were housewives, aged 17 to 30 years old, and having more than one child. They were educated to high school, knew some information on tetanus toxoid, and knew at least one of the tetanus symptoms. Almost all did not know the cause of tetanus. The logistic regression of knowledge on tetanus and TT immunization: mothers who heard of TT were 1.54 more likely to have been immunized than those who did not, while mothers who knew the use of tetanus toxoid were 2.15 times more likely to have been immunized than those who did not, and those who knew at least one of the tetanus symptoms were 1.86 times more likely to have been immunized than those who did not, respectively controlling other variables constant. Furthermore, women who had antenatal care were 30 times more likely to have been immunized than those who did not. Enhancing mothers knowledge on tetanus is important to increase the coverage of tetanus toxoid. Moreover, antenatal care would cause contact with sources of tetanus toxoid and hence increase the chance of having the immunization. At the same time, this decreases tetanus neonatorum. Considering the majority of pregnant women received information on tetanus from healthworkers, the use a variety of media would be advantageous.

[Epidemiological study of SMON death--from the observation of death certificates during a 13-year 6-month period from July 1972].

Eight hundred and twelve vital statistics death certificates, in which either the cause of death or complications was SMON, were selected from all the death certificates filed in Japan in a 13-year 6-month period from July 1972 to December 1985. In 411, or 50.6%, of these certificates, the SMON Research Committee had records of the clinical pictures as a result of the nationwide surveys conducted earlier. The epidemiological picture presented by the death certificates and the relationship between the information in these certificates and clinical records in nationwide surveys was observed. The results are summarized as follows: 1. The male/female ratio revealed in the 812 death certificates was 0.53, which is higher than those reported in nationwide surveys (0.4). 2. Although the yearly number of certificates was more than 70 in 1974 or before, it decreased year by year. The numbers in 1984 and 1985 were less than 50. 3. The leading underlying causes of death were; SMON in 44.0% of all the certificates, heart diseases in 10.3%, cerebrovascular diseases in 9.2% and malignant neoplasm in 8.6%. 4. The presence of clinical symptoms was compared between death cases and total cases reported by the nationwide surveys. Symptoms such as visual impairment and walking incapacity were seen more frequently in death cases. 5. From these results, it was suggested that the level of the severity of the disease at onset influences the long-term prognosis of the disease, although selection bias in the death certificates observed is unavoidable.

[Effects of topical prostaglandin analogues on the aqueous flare intensity in rabbit eyes at an early phase of endotoxin-induced uveitis].

PURPOSE: We examined the effects of prostaglandin analogues on the blood-aqueous barrier(BAB) permeability in rabbit eyes at an early phase of endotoxin-induced uveitis(EIU). SUBJECTS AND METHODS: One drop of 0.005% latanoprost or 0.12% unoprostone were applied to rabbit eyes. Escherichia coli lipopolysaccharides were injected to induce uveitis. The changes in flare intensity in normal eyes and EIU eyes after application of eye drops were evaluated. The effect of cyclooxygenase inhibitor on the flare intensity changes caused by the application of unoprostone was also examined. RESULTS: Flare intensity increased significantly after a single instillation of unoprostone, and the increase was not prevented by pretreatment with cyclooxygenase inhibitor. In eyes with EIU, unoprostone caused an additional increase of flare intensity to uveitis induced flare change. Latanoprost had no effects on BAB in eyes with normal and with uveitic conditions. CONCLUSION: Latanoprost and unoprostone did not cause an excessive inflammatory reaction in rabbit eyes at an early phase of EIU.

[Prevention of cerebro-cardiovascular diseases by early intervention in youth].

A significant number of youth have obesity, hypertension, hypercholesterolemia and diabetes mellitus which are major risk factors for C.V.D and I.H.D. and which frequently occur after maturity. Analysis of lifestyle factors show a strong relationship between a series of these factors, and life style practices as typified by time-saving, dietary practice such as convenience foods, insufficient exercises, as well as family history of C.V.D. Prompt establish next of a system for screening high risk children with these factors and a corresponding support system for guiding and instructing them, is indicated. Studies should be performed from the viewpoint of life-cycle health management for establishing an integrated examination system for circulatory diseases for screening to follow-up.

[A cohort study on deaths from SMON in Japan].

Causes and risk factors of deaths from subacute myelo-optico neuropathy (SMON) were studied in a prospective cohort of 4,329 SMON patients followed for 3 years and 7 months (Sept. 1985-March 1989) with the following findings: (1) Recent excess deaths of SMON patients was estimated as 4% from ratio of O/E (SMR = 104) and deaths due to SMON itself was 6.4%. (2) The ratio of O/E was significantly higher for deaths from cancer of colon/rectum in females, cancer of pancreas in males, hypertension in males, pneumonia/influenza in females, chronic obstructive pulmonary diseases in males, tuberculosis and intestinal obstructive disease in males and females. (3) The ratio of O/E was 1.8 times or greater for those SMON patients with complications of cerebrovascular disease, severe blindness, complete loss of ambulation, and who were bedridden, and who are unable to receive home care from family members or trained home helpers.

[Studies on the reduction and correction risk factors for arteriosclerosis in junior high school pupils].

Circulatory system examinations performed on first-grade pupils of junior high schools (12 years old; N = 1,204), identified 105 high-risk pupils with arteriosclerosis-promoting factors. These pupils were consulted on life-style modification, emphasizing nutrition and exercise, and encouraged to practice personal health management and self-monitoring. Reduction and correction of high risk factors were studied in these subjects, based on review of results of health examinations performed regularly up to the time of their graduation. Results are as follows: 1. Averages for obesity grade (for both sexes) in the second and the third years were significantly lower than that in the first year. Based on a general reference value > or = 20%, improvements were observed in 22.5% of males and 50.0% of females. 2. In both sexes average total cholesterol (TC) and triglyceride (TG) were lower in the second and the third years than in the first year, and a significant increase in HDL cholesterol was seen. With regard to a general reference value > or = 3.0 for AI (Atherogenic Index) calculated using TC and HDL cholesterol, improvements (reduction) were observed in 29.4%, of males and 46.2% of females. 3. While the numbers of subjects with elevated SBP, DBP, GPT, FPG and BUN were few, there were significant decreases in averages in the second and third years compared with those in the first year. Using general reference values, improvements in these risks were seen in 100% for all values except for GPT where 75.0% showed improvement. 4. Comparison of grouped distribution of summed values of examination results between the first year and the third year, showed improvements (reduction and correction of risks) in 69 pupils (65.7%), while 30 pupils (28.6%) remained in the same group with no improvement and 6 pupils (5.7%) turned for the worse, dropping one rank.(ABSTRACT TRUNCATED AT 250 WORDS)