Molecular genetic analysis of primary open-angle glaucoma, normal tension glaucoma, and developmental glaucoma for the VAV2 and VAV3 gene variants in Japanese subjects.

The VAV2 and VAV3 genes have been implicated as being causative for primary open angle glaucoma (POAG) in the Japanese. We studied 168 unrelated Japanese patients with primary open-angle glaucoma (POAG), 163 unrelated Japanese patients with normal tension glaucoma (NTG), 45 unrelated Japanese patients with developmental glaucoma (DG), and 180 ethnically matched normal controls, to determine whether variants in the vav 2 guanine nucleotide exchange factor (VAV2) and vav 3 guanine nucleotide exchange factor (VAV3) genes are associated with POAG, NTG, or DG in the Japanese. Genomic DNA was extracted from peripheral blood leukocytes, and variants in the VAV2 and VAV3 genes were amplified by polymerase chain reaction (PCR) and directly sequenced. Two variants were identified: rs2156323 in VAV2 and rs2801219 in VAV3. The variants and the prevalence of POAG, NTG, and DG in unrelated Japanese patients indicated that the variants were not involved in the pathogenesis of POAG, NTG, or DG.

Evaluation of CNTNAP2 gene polymorphisms for exfoliation syndrome in Japanese.

PURPOSE: To investigate the contactin-associated protein-like 2 (CNTNAP2) gene for single-nucleotide polymorphisms (SNPs) in Japanese patients with the exfoliation syndrome (XFS). METHODS: One hundred and eight unrelated Japanese patients with the XFS, and 199 normal controls were studied. Genomic DNA was extracted from the leukocytes of the peripheral blood, and 8 SNPs, rs826802, rs1404699, rs7803992, rs700308, rs4725736, rs2107856, rs2141388, and rs6970064, were amplified by polymerase chain reaction (PCR), directly sequenced, and genotyped. RESULTS: The allele frequencies of rs1404699 (p=8.57XE-3, odds ratio (OR)=1.59, 95% confidential intervals (CI); 1,12-2.24) and rs7803992 (p=5.43XE-4, OR=1.86, 95% CI; 1.31-2.65) were statistically significantly different between XFS and controls. In addition, there were significant differences in these genotype frequencies (p=0.0197 and 1.75XE-3). The allele and the genotype frequencies of rs2107856 and rs2141388, which were statistically significant SNPs in an earlier study, were not significantly different. CONCLUSIONS: The variants, rs1404699 and rs7803992, of CNTNAP2 should be associated with XFS in the Japanese population.

The relationship between advanced glycation end products and ocular circulation in type 2 diabetes.

AIMS: To determine whether skin autofluorescence (SAF) and serum pentosidine, biomarkers of advanced glycation end products (AGEs), were associated with ocular microcirculation in type 2 diabetes patients with early diabetic retinopathy (DR). METHODS: This study included 46 eyes of 46 type 2 diabetes patients with no DR or non-proliferative DR. SAF was measured with an autofluorescence reader. Optic nerve head (ONH) microcirculation, represented by mean blur rate (MBR), was measured with laser speckle flowgraphy. Overall MBR, vascular MBR, and tissue MBR were calculated in software. MBR, SAF, pentosidine levels, and clinical findings, including central macular thickness (CMT), were then compared. RESULTS: SAF in the diabetes patients was correlated with age (P=0.018). Serum pentosidine was correlated with age, vascular MBR and tissue MBR (P=0.046, P=0.035, and P=0.01, respectively). CMT was correlated with tissue MBR (P=0.016), but not with vascular MBR or overall MBR. Separate multiple regression analyses of independent contributing factors revealed that age, SAF, serum pentosidine, duration of diabetes, and pulse rate contributed to tissue MBR (P=0.041, P=0.046, P=0.022, P=0.011 and P=0.036, respectively), while SAF, HbA1c, pulse rate, tissue MBR, diastolic blood pressure, and creatinine contributed to CMT (P=0.005, P=0.039, P<0.001, P<0.001, P=0.022 and P=0.001, respectively). CONCLUSIONS: Tissue MBR may be closely related to AGE levels and CMT in type 2 diabetes patients with early DR, suggesting that ocular circulation might be potential early biomarkers of DR.

Diagnosing endometrial carcinoma with cervical involvement by cervical cytology.

OBJECTIVE: To assess the relationship of a cervical cytologic diagnosis based on number, size and degeneration of malignant clusters and necrotic background to cervical involvement of endometrial carcinoma. STUDY DESIGN: Cervical smears of 53 women with endometrial carcinoma were evaluated for cervical involvement. The cytologic diagnosis was compared with actual involvement, and accuracy was calculated. Retrospectively, cytologic features, including number, size and degeneration of malignant clusters and necrotic background, were analyzed in involved and noninvolved cases. RESULTS: Cervical involvement was confirmed in 15 patients (28.3%). The number and size of malignant clusters in the involved cases were significantly larger than those in the noninvolved cases (P < .001 and < .01, respectively). The proportion of degenerated malignant cells and necrotic background in involved cases were significantly higher than those in noninvolved cases (P < .05). Cytologic diagnosis had a sensitivity and specificity of 62.5% and 86.8%, respectively. CONCLUSION: Cervical smears of involved cases revealed a large number and large size of malignant clusters. These findings support cytologic diagnosis based on number, size and degeneration of malignant cells and necrotic background. Cervical cytology is useful to exclude cervical involvement because of its high specificity and can help detect cervical involvement because of its moderately high sensitivity.

[Seasonal allergic conjunctivitis in Hakodate].

PURPOSE: We investigated seasonal allergic conjunctivitis in Hakodate. MATERIALS AND METHODS: The clinical records of 293 patients with allergy-like symptoms who were seen at the Departments of Ophthalmology and Otolaryngology of Hakodate Municipal Hospital in 2001 and 2002 were examined for pollinosis, and evaluated. The general severity of conjunctivitis was graded as (+), (++), and (+++). The allergen was identified by AlaSTAT and nasal provocation test. RESULTS: In March, June, and September 2001, and in March, April, and June 2002, many patients were examined. In 2001, 134 patients (87.0%) had allergic conjunctivitis, and 126 patients (90.6%) had it in 2002. The severity of allergic conjunctivitis in May, June, and September was significantly (p<0.05) more severe than in February, March, and April. The major antigen rates for Cryptomeria japonica, Gramineae, Artemisia, and Betula platyphyia var. japonica were 22, 21, 8, and 3%. The specific IgE positive rates for Cryptomeria japonica, Betula platyphyia var. japonica, Gramineae and Artemisia were higher in March and April, in May, in June, and in September. CONCLUSION: The allergens of conjunctivitis in Hakodate were not the same as in the Kansai. We should consider the dispersion of pollens area by area.

Cervical cytology during pregnancy--comparison with non-pregnant women and management of pregnant women with abnormal cytology.

To clarify the clinical significance of uterine cervical cytology during pregnancy, we analyzed the incidence of cervical cytology and its accuracy. Of the 1,593 pregnant women underwent cervical cytology, the patients with abnormal cytology were followed up and performed histological confirmation on colposcopic biopsy specimen. An incidence of abnormal cytology and cervical neoplasm during pregnancy were 1.63% (26 cases) and 0.82% (13 cases), respectively. The incidence of abnormal cytology in the pregnant women was significantly higher than that (0.9%) in mass-screened, non-pregnant 214,375 women under the age of 45 years (P < 0.001). There was no significant difference of the incidence of cervical neoplasm between in the pregnant women and in mass-screened, non-pregnant women (0.82% vs. 0.46%). The accuracy of cervical cytology during pregnancy was 45.0% and this was not significantly different from that (27.6%) in the mass-screened, non-pregnant women. Since, cervical screening cytology for uterine cervical cancer in the pregnant women as shown in this study, has an equal effectiveness to that in the mass-screened non-pregnant women, routine cervical cytology is highly recommended to performed during pregnancy. In addition, management of pregnant women with abnormal cytology was discussed in this article.

[A study on the nasal and eye symptoms of pollinosis in hakodate].

We have investigated the nasal and eye symptoms of pollinosis in Hakodate. There were elevated levels of dispersion of Cryptomeria japonica pollen in Hakodate in 1995 and 1998. In 1995 and 2001, there were also elevated levels of dispersion of Betula platyphylia var. japonica pollen. From March to June and September from July 1999 to May 2002, there was an increased incidence of pollinosis in Hakodate. In 641 patients with allergic rhinitis, the specific IgE positive rates for house dust and mite, Artemisia (Art), Gramineae (Gra), Betulaplatyphylia var. japonica (BJ), and Cryptomeria Japonica (CJ) were 73.5, 28.5, 25.7, 14.2 and 21.2%, respectively. The subjects with pollinosis in 2001 and 2002 (n=95) were divided into four groups as follows: Cryptomeria japonica (n=49), Betula platyphylia var. japonica (n=11), Gramineae (n=18) and Artemisia (n=17) pollinosis. The general severity of rhinitis and conjunctivitis were divided into four grades ( (-), (+), (++) and (+++) ), respectively. The combined (++) and (+++) rates observed for the general severity of rhinitis for all these subjects (n=95) was 86.4% and there was no significant difference in the general severity of rhinitis between any of the four groups. The combined (++) and (+++) rates for the general severity of conjunctivitis for all subjects (n=95) was 56.9% and the general severity of conjunctivitis for BJ and Gra pollinosis were significantly (P<0.01) higher than those observed for CJ pollinosis. In conclusion, the present data shows that the general severity of rhinitis in subjects with pollinosis in Hakodate was more severe than that of conjunctivitis and also that the general severity of conjunctivitis was dependent on the specific pollen (s) responsible for the pollinosis.

Association of HK2 and NCK2 with normal tension glaucoma in the Japanese population.

Although family studies and genome-wide association studies have shown that genetic factors play a role in glaucoma, it has been difficult to identify the specific genetic variants involved. We tested 669 single nucleotide polymorphisms (SNPs) from the region of chromosome 2 that includes the GLC1B glaucoma locus for association with primary open-angle glaucoma (POAG) and normal tension glaucoma (NTG) in the Japanese population. We performed a two-stage case-control study. The first cohort consisted of 123 POAG cases, 121 NTG cases and 120 controls: the second cohort consisted of 187 POAG cases, 286 NTG cases, and 271 controls. Out of six SNPs showing significant association with POAG in the first round screening, seven SNPs were tested in the second round. Rs678350 in the HK2 gene coding sequence showed significant allelic (p=0.0027 in Stage Two, 2.7XE-4 in meta-analysis) association with POAG, and significant allelic (p=4.7XE-4 in Stage Two, 1.0XE-5 in meta-analysis) association with NTG. Although alleles in the TMEM182 gene did not show significant association with glaucoma in the second round, subjects with the A/A allele in TMEM182 rs869833 showed worse visual field mean deviation (p=0.01). Even though rs2033008 in the NCK2 gene coding sequence did not show significant association in the first round, it had previously shown association with NTG so it was tested for association with NTG in round 2 (p=0.0053 in Stage Two). Immunohistochemistry showed that both HK2 and NCK2 are expressed in the retinal ganglion cell layer. Once multi-testing was taken into account, only HK2 showed significant association with POAG and NTG in Stage Two. Our data also support previous reports of NCK2 association with NTG, and raise questions about what role TMEM182 might play in phenotypic variability. Our data suggest that HK2 may play an important role in NTG in the Japanese population.

Association of Toll-like receptor 4 gene polymorphisms in Japanese subjects with primary open-angle, normal-tension, and exfoliation glaucoma.

PURPOSE: To determine whether polymorphisms in the Toll-like receptor 4 (TLR4) gene are associated with primary open-angle glaucoma (POAG), normal-tension glaucoma (NTG), and exfoliation glaucoma (XFG) in Japanese individuals. DESIGN: Genetic association study. SETTING: Multicenter study. STUDY POPULATION: One hundred eighty-four unrelated Japanese patients with POAG, 365 unrelated patients with NTG, and 109 unrelated patients with XFG from 5 hospitals. PROCEDURES: Genomic DNA was extracted from leukocytes of the peripheral blood, and 8 polymorphisms in the TLR4 genes were amplified by polymerase chain reaction (PCR) and directly sequenced. Allele and genotype frequencies and the inferred haplotypes were estimated. MAIN OUTCOME MEASURES: Differences in allele and genotype frequencies and haplotypes between subjects with POAG, NTG, and XFG. RESULTS: The allele frequency of rs2149356 of the TLR4 gene in the POAG, NTG, and XFG groups was the most significantly different from that of the control group (minor allele frequency 0.446, 0.395, 0.404, vs 0.308; P = .000058, P = .0030, and P = .015). The allele frequencies of the 5 TLR4 SNPs were higher in all of the glaucoma groups than that in the control group. The statistics of genotypes of TLR4 were approximately the same for all allele frequencies. The haplotypic frequencies with Tag SNPs studied earlier showed that only POAG was statistically significant. Other haplotypes, such as rs10759930, rs1927914, rs1927911, and rs2149356, had higher statistical significance (overall P = .00078 in POAG, overall P = .018 in NTG, and overall P = .014 in XFG). CONCLUSIONS: This study demonstrated that TLR4 polymorphisms are associated with NTG in the Japanese, and they also play a role in the pathogenesis of POAG and XFG.

Irreversible optical clearing of rabbit dermis for autogenic corneal stroma transplantation.

Tissue engineering and transplantation of autogenic grafts have been widely investigated for solving problems on current allograft treatments (i.g., donor shortage and rejection). However, it is difficult to obtain an autogenic corneal stromal replacement that is composed of transparent, tough, and thick collagen constructs by current cell culture-based tissue engineering. Aim of this study is to develop transparent dermis for an autogenic corneal stroma transplantation. This study examined dehydration at 4-8°C and carbodiimide cross-linking on cloudy rabbit dermis (approx. 1.8%-3.8% light transmittance at 550 nm) for dermis optical clearing. Transparency of dehydrated rabbit dermis was founded to be approx. 37.9%-41.4% at 550 nm. Additional cross-linking treatment on dehydrated dermis prevented from swelling and clouding in saline, and improved its transparency to be 56.9% at 550 nm. Rabbit corneal epithelium was found to regenerate on optically cleared dermis in vitro. Furthermore, no abnormal biological response (i.e., inflammation, vascularization, and the barrier defect of epithelia) or no optical functional change on optically cleared dermis was observed during its 4-week autogenic transplantation into rabbit corneal stromal pocket.