Anti-tumor necrosis factor alpha antibody in the treatment of Behçet's disease.
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Adolescent↗
Biomedical subjects
Shigeaki Ohno
Publications and source records attributed to Shigeaki Ohno.
Expression of thymidine phosphorylase in choroidal malignant melanoma associated with neovascular glaucoma.
Reported herein is a case of 62-year-old man who complained of blurred vision and ocular pain in his right eye. The patient was diagnosed with choroidal melanoma complicated by neovascular glaucoma (NVG) and total retinal detachment, and he underwent enucleation of the eye. The isolated tumor was 2.5 x 2.5 cm in size. It was accompanied by intratumoral calcification, and consisted of epithelioid and spindle melanoma cells. There were a variety of microvessels in the stroma of the iris. The expression of thymidine phosphorylase (dThdPase), an angiogenic factor, was examined immunohistochemically. Cytoplasmic immunoreactivity for dThdPase was more prominent in the epithelioid cells than in spindle tumor cells. Another case of choroidal melanoma without NVG had less marked immunoreactivity. These results suggest that the production of dThdPase by melanoma cells correlates with the pathogenesis of NVG.
Biomarkers, Tumor↗
Five new genome types of adenovirus type 37 caused epidemic keratoconjunctivitis in Sapporo, Japan, for more than 10 years.
Human adenovirus type 37 (HAdV-37) is a major cause of epidemic keratoconjunctivitis and has recently been the largest causative agent of keratoconjunctivitis in Japan. To investigate the genetic characteristics of HAdV-37 strains isolated in Sapporo, we analyzed the genome types and genetic relationships of 51 strains isolated there from 1990 through 2001. By using DNA restriction analysis, eight genome types (HAdV-37/D1, HAdV-37/D3, and HAdV-37/D6 to HAdV-37/D11) were identified, including five new ones. The restriction fragments of these genome types shared more than 95% identity with those of the prototype strain. By DNA sequence analysis, five and three single nucleotide substitutions, respectively, were found in partial sequences of the hexon and fiber genes. The combinations of mutations resulted in four hexon and fiber types (hx1 to hx4 and f1 to f4) and six hexon/fiber pairs (hx1/f1, hx2/f1, hx1/f2, hx1/f3, hx3/f4, and hx4/f4). The six pairs correlated well with certain genome types. In all three epidemics of keratoconjunctivitis to strike Sapporo in the past 12 years, specific genome types and fiber types were usually isolated: in the first epidemic, HAdV-37/D1 (f1) and HAdV-37/D3 (f1); in the second, HAdV-37/D6 (f2) and HAdV-37/D8 (f3); and in the third, HAdV-37/D10 (f4) and HAdV-37/D11 (f4). We conclude that mutations in the adenovirus genome occurred chronologically and that certain mutations were correlated with the epidemics of adenoviral keratoconjunctivitis.
Adenovirus Infections, Human↗
Sensitive and rapid detection of herpes simplex virus and varicella-zoster virus DNA by loop-mediated isothermal amplification.
Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid amplification method in which reagents react rapidly and efficiently, with a high specificity, under isothermal conditions. We used a LAMP assay for the detection of herpes simplex virus type 1 (HSV-1), herpes simplex virus type 2 (HSV-2), and varicella-zoster virus (VZV). The virus specificities of primers were confirmed by using 50 HSV-1, 50 HSV-2, and 8 VZV strains. The assay was performed for 45 min at 65 degrees C. The LAMP assay had a 10-fold higher sensitivity than a PCR assay. An analysis of nucleotide sequence variations in the target and primer regions used for the LAMP assay indicated that 3 of 50 HSV-1 strains had single nucleotide polymorphisms. No HSV-2 or VZV strains had nucleotide polymorphisms. Regardless of the sequence variation, there were no differences in sensitivity with the HSV-1-specific LAMP assay. To evaluate the application of the LAMP assay for clinical diagnosis, we tested clinical samples from 40 genital herpes patients and 20 ocular herpes patients. With the LAMP assay, 41 samples with DNA extraction and 26 direct samples without DNA extraction were identified as positive for HSV-1 or HSV-2, although 37 samples with DNA extraction and just one without DNA extraction were positive by a PCR assay. Thus, the LAMP assay was less influenced than the PCR assay by the presence of inhibitory substances in clinical samples. These observations indicate that the LAMP assay is very useful for the diagnosis of HSV-1, HSV-2, and VZV infections.
Animals↗
Anti-inflammatory effects of aronia extract on rat endotoxin-induced uveitis.
PURPOSE: Aronia crude extract (ACE) with high levels of polyphenol compounds has been reported to have antioxidative effects in vitro and in vivo. In this study, attention was focused on the antioxidant effect of ACE. The purpose of the present study was to investigate the effect of ACE on endotoxin-induced uveitis (EIU) in rats. In addition, the endotoxin-induced expression of the inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 proteins was investigated in a mouse macrophage cell line (RAW 264.7) treated with ACE in vitro, to clarify the anti-inflammatory effect. METHODS: EIU was induced in male Lewis rats by a footpad injection of lipopolysaccharide (LPS). Immediately after the LPS inoculation, 1, 10, or 100 mg ACE or 10 mg prednisolone was injected intravenously. After 24 hours, the aqueous humor was collected from both eyes, and the number of infiltrating cells, protein concentration, nitric oxide (NO), prostaglandin (PG)-E2, and TNF-alpha levels in the aqueous humor were determined. RAW 264.7 cells treated with various concentrations of ACE were incubated with 10 mug/mL LPS for 24 hours. Levels of NO, PGE2, and TNF-alpha were determined by an enzyme-linked immunosorbent assay. The expression of iNOS and COX-2 proteins was analyzed by Western blot analysis. RESULTS: The number of inflammatory cells, the protein concentrations, and the levels of NO, PGE2, and TNF-alpha in the aqueous humor in the groups treated with ACE were significantly decreased in a dose-dependent manner. In addition, the anti-inflammatory effect of 100 mg ACE was as strong as that of 10 mg prednisolone. The anti-inflammatory action of ACE was stronger than that of either quercetin or anthocyanin administered alone. ACE also suppressed LPS-induced iNOS and COX-2 protein expressions in RAW 264.7 cells in vitro in a dose-dependent manner. CONCLUSIONS: The results suggest that ACE has a dose-dependent anti-ocular inflammatory effect that is due to the direct blocking of the expression of the iNOS and COX-2 enzymes and leads to the suppression of the production of NO, PGE2, and TNF-alpha.
Animals↗
Role of neurotrophin-4/5 in neural cell death during retinal development and ischemic retinal injury in vivo.
PURPOSE: Neurotrophin (NT)-4/5 and brain-derived neurotrophic factor (BDNF) mediate cell survival through TrkB, a high-affinity tyrosine kinase receptor, and may prevent neural cell death in various pathologic conditions. This study was conducted to investigate the function of NT-4/5 in neural cell death during retinal development and ischemic retinal injury. METHODS: Retinal development in wild-type, NT-4/5 knockout (KO), and NT-4/5:BDNF double-KO mice was histologically examined from postnatal day 0 (P0) to P90. Ischemic retinal injury was performed at P42, and NT-4/5 mRNA expression level and the extent of retinal cell death was quantitatively examined. RESULTS: Real-time PCR analysis revealed increased NT-4/5 mRNA expression in the ischemic retina. In the NT-4/5 KO mouse, retinal development and structure were normal, but the strain was susceptible to ischemic injury on P42. In contrast, NT-4/5:BDNF double-KO mice showed delayed retinal development and died before P42. CONCLUSIONS: These results suggest that NT-4/5, in combination with other trophic factors, is involved in the postnatal survival of retinal neurons during both development and degeneration.
Animals↗
[Behçet's disease].
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Amino Acid Sequence↗
Phosphorylation of p27(KIP1) in the developing retina and retinoblastoma.
Cellular distribution of the p27(KIP1) protein and its phosphorylation on threonine (T) 187 in mouse retinas from three stages of development, and retinoblastoma were examined. Retinas in C57Bl6 mice at embryonic day (E) 14, postnatal day (P) 1 and P11 were analyzed using immunohistochemistry with anti-p27(KIP1), threonine-187-phosphorylated p27(KIP1) (T187-phospho-p27), bromodeoxyuridine (BrdU), proliferating cell nuclear antigen (PCNA) antibodies, and phosphorylated histon H3 (pHiston H3), which is a marker for cells in M phase. p27(KIP1) knockout (-/-) mice and human retinoblastoma were also analyzed. T187-phospho-p27 was detected in the outermost layer of the retina, whereas several neuroblastic cells expressed p27(KIP1) at E14. Many neuroblastic cells expressed BrdU in the middle layer. At P1, p27(KIP1) was detected in the ganglion cell layer and neuroblastic layer. T187-phospho-p27 was detected in the outermost layer, and that was localized in mitotic cells that also showed pHiston H3-positive. At P11, p27(KIP1) was detected in the inner nuclear layer, whereas T187-phospho-p27-positive or mitotic cells were not. BrdU positive nuclei were not detected in wild-type but were noted in the inner nuclear layer and the outer nuclear layer of the p27(KIP1) -/- mice retina at P11. In retinoblastoma, tumor cells formed numerous rosettes with Flexner-Wintersteiner rosettes. Several pHiston H3 -positive nuclei were noted in the tumor cells forming Flexner-Wintersteiner rosettes. Several T187-phospho-p27-positive nuclei were also detected in the mitotic cells forming Flexner-Wintersteiner rosettes. PCNA was expressed in rosette-forming cells. In conclusion, T187-phospho-p27(KIP1) was correlated with M phase of the cell cycle in the developing retina and retinoblastoma.
Animals↗
[Clinical and virological studies of nosocomial conjunctivitis infection caused by adenovirus type 37 variant].
PURPOSE: To study the clinical features and virological analysis of the nosocomial adenoviral conjunctivitis cases occurring in the ophthalmology ward of Fukushima Medical University Hospital. MATERIALS AND METHODS: We studied the symptoms and clinical course of 61 patients who had adenoviral conjunctivitis caused by nosocomial infections in our hospital. We attempted to detect the adenovirus antigen, analyze the viral DNA, and isolate the virus from conjunctival swabs. RESULTS: The clinical symptoms of adenoviral conjunctivitis were mainly conjunctival hyperemia, discharge and conjunctival follicles. Adenoviral conjunctivitis patients who had undergone ophthalmic surgery had conjunctivitis in the operated eye. The sensitivity of Adeno-check was 78.9% in the in-patients. Adenovirus type 37 variant was detected by molecular analysis and viral isolation. CONCLUSIONS: Adenoviral conjunctivitis can often lead to outbreaks of nosocomial infection in the ophthalmic ward and sometimes requires makes necessary restriction of hospitalization and closing of the ward. Therefore, patients need to be observed carefully. The virological analysis of specimens from conjunctival swabs detected pathogens and provided useful information concerning adenoviral conjunctivitis.
Adenoviruses, Human↗
[Studies on exogenous and endogenous factors associated with various ocular diseases].
The exact cause and nature of various ocular diseases are still unknown. In accordance with the progress of modern medical science, technology for research investigations has greatly developed and a variety of new results have recently been produced. However, they were mostly about exogenous factors such as bacteria or parasites. On the other hand, molecular biology has markedly progressed in the second half of the 20th century and has been applied to medical science. As for the exogenous factors, molecular techniques have been introduced to the diagnosis of infectious diseases, and their diagnosis has become much easier and much more exact than before. Similarly, in the field of immunogenetics, molecular genetics has greatly developed, and disease susceptibility or disease resistance has been investigated at the DNA or molecular level. Regarding the exogenous disease mechanisms, most infectious diseases occur when the microorganisms are transmitted to some individuals. In this case, endogenous factors play almost no or very little role. Adenovirus keratoconjunctivitis is one example. Human adenovirus (HAdV) is a DNA virus. Although HAdVs are classified into 51 different serotypes, only 7 of them such as HAdV-3, -4, -7, -8, -11, -19, and -37 usually develop acute keratoconjunctivitis. The genome of HAdV develops constant mutation, and new genome types often cause epidemics of nosocomial infections. Recent studies showed that most of the keratoconjunctivitis cases are caused by HAdV-37 in Japan. With regard to the endogenous disease mechanisms, only 30 years have passed since the recent molecular genetic studies on disease susceptibility started. Before that, genetic disease mechanisms were studied only in hereditary diseases such as color vision defects, which follow the modes of Mendelian inheritance. Therefore, the presence of disease susceptibility was not clearly understood in the so-called nonhereditary diseases. However, after our initial findings of the close association between Behçet's disease and HL-A 5 (later HLA-B 5 or HLA-B 51) in 1973, there were many reports on the presence of genetic predisposition to certain non-hereditary diseases. One example is Vogt-Koyanagi-Harada's (VKH) disease which has a close association with HLA-DRB 1*0405 in Japanese patients, as also was the case with sympathetic ophthalmia. These studies indicate that Behçet's disease may be called "a disease of the Yayoi people" and Vogt-Koyanagi-Harada's disease as "a disease of the Jomon people" when analyze the associated genetic factors. On the other hand, the human genome project has been successfully completed and it is now possible for us to investigate any genetic factor associated with certain diseases either on 22 pairs of autosomal chromosomes or on one pair of sex chromosomes. Our collaborative team has adopted the microsatellite methods utilizing the pooled DNA PCR method, and started investigation of any genetic association with ocular diseases, in addition to the HLA system on the short arm of autosomal chromosome 6. In the first screening of 100 DNA samples of Behçet's disease patients, for example, 9 % of microsatellite markers on the autosomal chromosomes 1, 6, 17, and 19 showed significant positive association. The same studies are now in progress by our collaborative team on inflammatory diseases such as sarcoidosis, and on noninflammatory conditions like hypertension or high myopia. These results suggest that exogenous environmental factors such as infections which may trigger the occurrence of the disease or conditions, and endogenous genetic factors which may predispose someone to contracting a disease, are both significantly associated with various ocular diseases, and construct intricate disease mechanisms. Therefore, in future studies of diagnosis, treatment, and prevention of various ocular diseases, both exogenous factors and endogenous factors have to be carefully analyzed and investigated.
Adenoviridae↗
Expression of maf-B mRNA in the epithelium around the eyelid closure of the mouse eye at embryonic day 18.
Maf encodes a transcription factor protein containing a typical basic leucine zipper domain structure, a motif for protein dimerization and DNA binding. We examined the expression of maf-B mRNA in the epithelium around the eyelid closure. Expression of maf-B mRNA was examined in C57Bl6 mice at the embryonic stages in 12.5 days of gestation (E12.5) and E18 using in situ hybridization with 35S-labeled antisense riboprobes. In embryos studied 12.5 days postconception, a message specific for maf-B was not detected around the developing eyelid. In contrast, maf-B was strongly expressed in the epithelium of the eyelid closure at E18. Expression of maf-B was strongly noted in the suprabasal differentiating cells derived from the basal layer of the conjunctiva and epidermis. In contrast, basal cells in the eyelid closure and in the epidermis, as well as keratinizing cells, did not express maf-B. These data indicate that maf-B mRNA is expressed during development of the eyelid closure.
Animals↗
A patient with sarcoidosis diagnosed by a biopsy of scleral nodules.
PURPOSE: To report the case of a patient with nodular scleritis who was diagnosed as having sarcoidosis by a biopsy of scleral nodules. METHODS: Case report. RESULTS: A 26-year-old man with nodular scleritis underwent a biopsy of scleral nodules. The year before, he had been diagnosed with granulomatous iridocyclitis of the right eye. Although sarcoidosis had been suspected, extensive systemic examinations resulted in no positive findings of sarcoidosis. Histopathological specimens revealed non-caseating epithelioid granuloma with giant cells. The nodular scleritis was resolved by topical corticosteroids. CONCLUSIONS: A scleral biopsy is useful for the diagnosis of sarcoidosis when the patient has nodular scleritis with no systemic signs.
Adult↗
Distribution of p27(KIP1), cyclin D1, and proliferating cell nuclear antigen after retinal detachment.
PURPOSE: To examine the expression of the p27(KIP1), cyclin D1, and proliferating cell nuclear antigen (PCNA) in the retina and retinal pigment epithelium (RPE) after retinal detachment. METHODS: Normal eyes and eyes at 2 or 4 days after retinal detachment with the C57B16 mouse were analyzed by immunocytochemistry using anti-p27(KIP1), anti-cyclin D1, and anti-proliferating cell nuclear antigen (PCNA) antibodies as well as anti-glutamate synthetase (GS) antibody. RESULTS: The p27(KIP1) positive nuclei were distributed in the inner nuclear layer (INL) and the RPE of the normal mice eye. In the INL, p27(KIP1) was detected in the middle sublayer, where the nuclei of glutamate synthetase positive Müller cells were situated. In contrast, cyclin D1 was not detected either in the retina or in the RPE. At 2 and 4 days after the retinal detachment, RPE cells under the detached retina were negative for p27(KIP1) and positive for cyclin D1 and PCNA. In the INL of the detached retina, p27(KIP1) was detected after 2 days, but was not detected after 4 days. In contrast, PCNA was not detected in the INL after 2 days, but was detected after 4 days. Cyclin D1 was detected in the middle sublayer of the INL at both 2 and 4 days after the retinal detachment. CONCLUSION: These results suggested that degradation of p27(KIP1) and expression of cyclin D1 was involved in the proliferation of the Müller cells as well as RPE cells after retinal detachment.
Animals↗
Involvement of p27(KIP1) in proliferation of the retinal pigment epithelium and ciliary body.
According to observations in various cell lines, elimination of the cyclin-dependent kinase inhibitor p27(KIP1) during the late G1 phase of the cell cycle is required for progression to the S phase. Eyes from C57BL/6 mice at embryonic days 13, 14, and 18, and at 4 weeks of age, were analyzed by a bromodeoxyuridine cell proliferation assay and by immunocytochemistry using anti-p27(KIP1) antibody. On embryonic days 14 and 18, p27(KIP1) was detected in the ciliary body. This protein also was detected in the nuclei of the many cells of the retinal pigment epithelium on embryonic day 18, and was present in all such cells at 4 weeks of age. When p27(KIP1)-/- knockout and control mice were injected with bromodeoxyuridine between postnatal days 7 and 10 and analyzed on day 11, positive cells were abundant in the retinal pigment epithelium and the ciliary body of p27(KIP1)-/- mice, whereas few cells were positive in control mice. By fluorescent nuclear staining in whole mounts of retinal pigment epithelium at 12 weeks of age, more nuclei were present in p27(KIP1)-/- than in the wild-type mice. These results suggest that p27(KIP1) was involved in regulation of proliferation in the RPE and the ciliary body.
Animals↗
Association between IFNA genotype and the risk of sarcoidosis.
Sarcoidosis is known to be a systemic granulomatous disorder characterized by a cell-mediated Th1-type inflammatory response. To identify a key genetic factor in the pathogenesis of sarcoidosis, we investigated single nucleotide polymorphisms within 10 candidate genes involved in type 1 immune process ( IFNA17, IFNB, IFNG, IFNGR1, IFNGR2, IL12B, IL12RB1, IL12RB2, ETA-1, and NRAMP1) in an association-based study of 102 Japanese patients with sarcoidosis, 114 with tuberculosis, and 110 control subjects. After correction for multiple testing, an IFNA17 polymorphism (551T-->G) was found to be associated with susceptibility to sarcoidosis (odds ratio 3.27 [95% CI: 1.44-7.46], P=0.004, P(c)=0.04), but not to tuberculosis. We observed no significant associations with the other polymorphisms of the Th1-related genes. We further typed another IFNA polymorphism ( IFNA10 60T-->A) and confirmed two major haplotypes of the IFNA gene, viz., allele 1: IFNA10 [60T]- IFNA17 [551T] and allele 2: IFNA10 [60A]- IFNA17 [551G], in the Japanese population. In healthy subjects, IFNA allele 2, which is over-represented in patients with sarcoidosis, was significantly associated with increased IFN-alpha and IL-12p70 production induced by Sendai virus in vitro. This study suggests that possession of the IFNA allele with higher levels of IFN-alpha significantly increases the risk of sarcoidosis.
Adolescent↗
Diverse NF-kappaB expression in epiretinal membranes after human diabetic retinopathy and proliferative vitreoretinopathy.
PURPOSE: Formation of epiretinal membranes (ERMs) after proliferative diabetic retinopathy (PDR) and proliferative vitreoretinopathy (PVR) results in progressive deterioration of vision, but its pathogenic mechanisms are still unknown. This study was conducted to examine the role of nuclear factor kappa B (NF-kappaB) in the formation of ERMs after PDR and PVR. METHODS: ERM samples were obtained by vitrectomy from 10 patients with PDR (aged 53+/-12 years with 14+/-5 years of diabetes), 20 patients with PVR, and 17 patients with idiopathic ERMs. Ten PVR and 17 idiopathic ERM samples were processed for reverse transcription-polymerase chain reaction (RT-PCR) analysis. In addition, 10 PDR and 10 PVR membranes were processed for immunohistochemical analysis. RESULTS: NF-kappaB mRNA expression levels were significantly higher (10 of 10 versus 9 of 17 subjects in idiopathic ERM, p=0.0119) in PVR subjects. Immunohistochemical analysis showed NF-kappaB protein expression in 8 of the 10 PDR samples as well as all 10 PVR samples, and NF-kappaB positive cells were partially double labeled with glial cell markers. Interestingly, NF-kappaB protein was also overlapped with angiogenic factor interleukin-8 (IL-8) in glial cells as well as vascular endothelial cells. CONCLUSIONS: These results suggest that NF-kappaB is involved in the formation of both glial and vascular endothelial cell components, and that these two cell types might have functional interactions that lead to the enlargement of intraocular proliferative membranes.
Adult↗
The cotton rat model for adenovirus ocular infection: antiviral activity of cidofovir.
To determine the antiviral effects of compounds against ocular adenovirus (AdV) infection, we established an animal model of AdV infection in cotton rat eyes. Cotton rat eyes were inoculated intrastromally and topically with four AdV serotypes 4, 5, 8, and 37, and treated topically with 1% HPMPC (cidofovir) eye drops twice a day. The infected corneas were extracted and homogenized, and virus titers in the cornea specimens were determined by a plaque assay. The virus titer in AdV type 5-inoculated eyes peaked on days 0 through 3 after inoculation and virus shedding was detected for 18.0+/-2.8 days. AdV 5 antigen in the infected corneas was demonstrated in the corneal epithelial cells by immunofluorescence stain. However, for AdV serotypes 4, 8, and 37, no evidence of continued virus replication in cotton rat eyes was noted. Specimens from cidofovir-treated eyes infected with AdV 5 demonstrated a statistically significant reduction in the mean virus titer (days 3-15) (P=0.028) and virus shedding duration (P=0.0014), as compared with those of the control group.
Adenoviridae Infections↗
The effects of Ginkgo biloba extract on lipopolysaccharide-induced inflammation in vitro and in vivo.
PURPOSE: Ginkgo biloba extract (GBE) contains many different flavone glycosides and terpenoides. Several previous studies have demonstrated that GBE exhibits a wide variety of biological activities, including an antioxidant action, on which we focused our attention. The aim of the present study was to investigate the efficacy of GBE on endotoxin induced uveitis in rats. The anti-inflammatory potency of GBE in vivo was compared with that of prednisolone. In addition, we also investigated nitric oxide (NO), prostaglandin E2 (PGE2), tumor necrosis factor-alpha (TNF-alpha) and the expression of iNOS in a mouse macrophage cell line (RAW 264.7) treated with GBE in vitro to clarify the anti-inflammatory effect. METHODS: EIU was induced in male Lewis rats by a footpad injection of lipopolysaccharide (LPS). Immediately after the LPS inoculation, either 1, 10 or 100 microg of GBE were injected intravenously. 24hr later, the aqueous humor was collected from both eyes, and the number of infiltrating cells, protein concentration and NO level in the aqueous humor was determined. The RAW 264.7 cells were pretreated with various concentrations of GBE for 24hr and subsequently incubated with LPS for 24hr. Levels of NO, PGE2 and TNF-alpha were determined by enzyme-linked immunosorbent assay. The expression of iNOS protein was analyzed by Western blotting method. RESULTS: GBE treatment in vivo decreased the concentrations of protein and NO in the aqueous humor of EIU rats. The anti-inflammatory effect of 1 mg GBE was as strong as that of same dose prednisolone. It also significantly reduced the concentration of PGE2, TNF-alpha and NO production in the medium of RAW 264.7 cells compared to that of the LPS group in vitro. The expression of iNOS protein in the 1000 microg ml(-1) of GBE treated cells decreased significantly. CONCLUSION: The present results indicate GBE suppresses the inflammation of EIU by blocking the iNOS protein expression and its anti-inflammatory effect on eye is comparable with the effect of prednisolone used in similar doses.
Animals↗