[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.

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.

The human behavioral and socioeconomic determinants of malaria in Bacan Island, North Maluku, Indonesia.

In eastern Indonesia, malaria control activities mainly depend on residual spraying but the situation is almost unchanged since the past decade. Understanding the socioeconomic and human behavior determinants is needed to implement an effective malaria control in accordance with the local condition and development. Hence we conducted an unmatched case control study. Two hundred samples were recruited from all, 11 villages surrounding the centre in Bacan Island, Maluku. For children aged 0 to 15 years old, the association of socioeconomic determinants: crowding and poor type of houses with malaria remained significant in the multivariate analysis. Meanwhile for persons above 15 years old, younger persons and regular going outside at night remained significant in the multivariate analysis. And for persons above 15 years old, a higher proportion of controls (14%) than cases (4%) slept under mosquito net regularly. The Indonesia Family Program should be promoted. There was a better quality of life in small family. For persons above 15 years old, going outside at night should be discouraged because exposed to mosquito bites. The malaria control strategy use of effective personal, regular use of mosquito net could be used as a completion for the present activities. Considering the low malaria knowledge among samples, inhabitants should be enhanced the malaria knowledge on causation, transmission, prevention and to provide proper knowledge on residual spraying.

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.

Molecular analysis of familial androgen insensitivity syndrome due to replacement of glutamic acid 802 by lysine.

We studied a Japanese family presenting at least two cases of complete androgen insensitivity syndrome (CAIS) and negative androgen receptor binding. The index subject showed a 46, XY karyotype and a complete female phenotype. For the purpose of further diagnosis and genetic counseling, molecular analysis of the androgen receptor (AR) gene was performed. Direct sequencing of the AR gene identified a mutation at nucleotide 2935 (A-->G). This replacement was a novel missense mutation, resulting in the substitution of glutamic acid 802 by lysine which deleted a recognition site for EcoRI in exon 6 of the AR gene. We identified another affected individual, using chromosome and molecular analysis of the AR gene at exon 6. Furthermore, although heterozygote carriers could not be identified on clinical grounds, molecular identification of healthy individuals and heterozygote carriers in the family members provided definitive information for genetic counseling. We believe that the molecular analysis of familial CAIS is very informative for both the affected individuals and other family members.

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 comparison between the nationwide epidemiological survey in 1991 and previous surveys on behçet's disease in Japan.

The 4th nationwide epidemiological survey on Behçet disease (BD), which included all patients with BD at 1,200 hospitals selected at random from 10,081 hospitals in Japan, was carried out by the BD Research Committee of the Ministry of Health and Welfare in 1991 to examine the epidemiological features of BD in Japan by comparing with previous surveys. 3,938 patients from these hospitals were examined by the Japanese diagnostic criteria of BD (JCBD) revised in 1987 and the International criteria for classification of BD (ICBD). Among these 3,938 patients, 622 patients were only suspected of having BD or clinical signs of the disease were unknown, and most of these patients were incompatible with the ICBD. So these patients were excluded from the study of epidemiological features. The average patients age has risen 7-8 years over the last 20 years and the average age of onset in both sexes increased by about 3 years from 1972 to 1991. While a decrease in the sex ratio was seen in the complete-type and the incomplete-type BD without ocular symptoms, a sustained high sex ratio was shown in incomplete-type BD with ocular symptoms. The positive rate of HLA-B51 antigen was 54.9% (men: 56.9%, women: 52.2%) significantly higher than die 15-16% in healthy subjects but it might have been gradually decreasing. Also the clinical course of BD has become too mild for prognosis. According to diese epidemiological features of BD, the clinical manifestation of BD in Japan might have become the Western type of BD.

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.

A central role for Stat3 in IL-6-induced regulation of growth and differentiation in M1 leukemia cells.

Interleukin-6 (IL-6) induces either differentiation or growth of a variety of cells. Little is known about the molecular basis of this cellular decision. The family of signal transducer and activator of transcription (Stat) proteins are involved in signaling through a variety of cytokine and growth factor receptors, although their biological roles have not been established. To address whether Stat proteins play roles in IL-6-induced growth or differentiation, we introduced two types of mutant Stat3 acting in a dominant-negative manner into M1 leukemic cells which respond to IL-6 with growth arrest and terminal differentiation. We show that dominant-negative forms of Stat3 inhibited both IL-6-induced growth arrest at G(0)/G1 and macrophage differentiation in the M1 transformants. Blocking of Stat activation resulted in inhibition of IL-6-induced repression of c-myb and c-myc. Furthermore, IL-6 enhanced the growth of M1 cells primarily through shortening the length of the G1 period when Stat3 was suppressed. Thus IL-6 generates both growth-enhancing signals and growth arrest- and differentiation-inducing signals at the same time. Stat3 may be a key molecule which determines the cellular decision from cell growth to differentiation in M1 cells.

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.

ERM, a PEA3 subfamily of Ets transcription factors, can cooperate with c-Jun.

A human cDNA clone for ERM, a member of the ets gene family, has been obtained by polymerase chain reaction with degenerate primers corresponding to highly conserved regions within an Ets DNA binding domain. ERM mRNA is expressed ubiquitously. The gene was mapped to chromosome 3q27. In in vivo transient-expression assays, ERM induced transcription more efficiently from a synthetic element containing both an ets-binding site (EBS) and a cyclic AMP response element (CRE) than from one containing an EBS alone. The activation of a synthetic EBS-CRE site by ERM was likely to involve a leucine zipper protein capable of dimerizing with CRE-BP1 leucine zipper. Indeed, ERM and c-Jun synergistically activated the EBS-CRE without making an apparent ternary complex. The synergy between c-Jun and ERM may be attributed to the enhancing effect of c-Jun on the transcription activity of ERM, because c-Jun increased ERM transcription activity by more than 20-fold in an assay system using a variety of fusion proteins between a Gal4 DNA-binding domain and a portion of ERM. This enhancing effect of c-Jun required the amino-terminal portion of ERM.

Effects of endurance training on gene expression of insulin signal transduction pathway.

Alternative splicing of insulin receptor mRNA and gene expression of insulin receptor, IRS-1 and MAP kinase isoforms were examined in skeletal muscle of trained and sedentary rats. Adult male Sprague-Dawley rats were trained for 9 weeks on a treadmill: 30 m/min at 6 degrees incline, 90 min/day, 5 days/week. Endurance training increased insulin receptor mRNA level without change in alternative splicing of insulin receptor mRNA in skeletal muscle. The levels of IRS-1 and MAP kinase (ERKI) mRNA were significantly higher in trained rats than sedentary rats. Our findings provide the first evidence that gene expression of insulin receptor and postreceptor signal transduction pathway is enhanced by endurance training, without affecting alternative splicing of insulin receptor isoforms.

Transcriptional activation of the IL-6 response element in the junB promoter is mediated by multiple Stat family proteins.

IL-6 signals activate an IL-6 response element in the junB promoter, JRE-IL6, in a Ras-independent manner. IL-6 rapidly induced a DNA-binding activity to the Ets binding site of JRE-IL6 (JEBS), one of necessary DNA motifs of the IL-6 response element. The IL-6-induced JEBS-binding activity was indistinguishable from those to acute phase response element (APRE) in both kinetics of induction and its DNA-binding specificity. Purified APRE binding factors (APRFs) from IL-6-stimulated rat liver were found to be composed of multiple Stat3-related proteins and Stat1. Moreover the purified APRFs specifically made a complex with JRE-IL6 with the same mobility as that observed in the crude extracts. These results indicate that an immediate early signal of IL-6 leading to activation of JRE-IL6 is mediated by STAT family transcription factors.

[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)

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.