Molecular characterization in the VP7, VP4 and NSP4 genes of human rotavirus serotype 4 (G4) isolated in Japan and Kenya.

The VP7, VP4 and NSP4 genes of human rotavirus serotype 4 (G4) were analyzed to investigate intraserotypic variations. The techniques used included reverse transcription polymerase chain reaction with subtype specific primers, restriction fragment length polymorphism analysis and sequence analysis. Twelve isolates (nine from Japan and three from Kenya) and two standard strains (Hochi, Odelia) were G4A P[8] Wa group NSP4. A standard strain (ST3) was G4A P[6] Wa group NSP4 and a strain (VA70) was G4B P[8] Wa group NSP4. These results show G4 rotaviruses can be divided into three combinations at the moment.

[Visual outcome of macular hole surgery with internal limiting membrane peeling].

PURPOSE: To evaluate the efficacy of the internal limiting membrane(ILM) peeling on macular hole surgery. METHODS: A series of 102 patients(105 eyes) who underwent primary macular hole surgery between October 1994 and April 1999 was used for this retrospective study. The mean age was 65.6 +/- 7.1 years (mean +/- standard deviation). Of the study eyes, 34 eyes(32%) had a Stage II hole, 49 eyes(47%) had a Stage III hole, and 22 eyes(21%) had a Stage IV hole based on the Gass classification. Here we compared the surgical and visual outcome of the ILM peeling-treated group(treated group: 51 eyes) with those of ILM peeling-untreated group(untreated group: 54 eyes). RESULTS: The hole closure rate after initial surgery was 98.0% in the treated group and 90.7% in the untreated group, and mean postoperative visual acuity, excluding cases where the hole was not closed by initial surgery, was 0.44 and 0.47, respectively. Visual improvement of 2 or more lines on Snellen chart was achieved in 84.3% and 57.4%(p < 0.01), and that of 4 or more lines in 54.9% and 25.9% (p < 0.01), respectively. Of the eyes with Stage II and III holes, visual outcome of the treated group was significantly better than that of the untreated group(p = 0.034, p = 0.037). In Stage IV, the initial closure rate of the treated group was significantly better than that of the untreated group(p = 0.02), but the visual outcome was not significantly different. CONCLUSION: Vitreous surgery combined with ILM peeling for the management of idiopathic macular hole is effective not only on hole closure but also on visual recovery.

Two-year follow-up study comparing primary vitrectomy with scleral buckling for macula-off rhegmatogenous retinal detachment.

PURPOSE: To compare the anatomic and visual outcomes achieved by scleral buckling and primary vitrectomy for the repair of macula-off rhegmatogenous retinal detachment. METHODS: The records were reviewed for a consecutive series of 167 patients (167 eyes) who were initially treated with scleral buckling or pars plana vitrectomy for primary macula-off retinal detachment. Patients were treated between January 1993 and December 1996. After adjustments for preoperative characteristics, data from 102 cases (55 scleral buckle cases and 47 primary vitrectomy cases) were used for the final comparison. There had been a minimum follow-up period of 24 months. RESULTS: No significant differences in single-procedure reattachment incidence (91%), final success incidence (100%) and incidence of postoperative proliferative vitreoretinopathy development (4%) were observed between the two treatment groups. Preoperative visual acuity, preoperative intraocular pressure, and duration of macular detachment were the three best predictors of postoperative visual recovery in both groups. Favorable overall visual recovery was obtained postoperatively, with no significant differences between the two groups throughout the follow-up period. However, in the eyes with poor preoperative visual acuity (<0.1), ocular hypotony (intraocular pressure <7 mm Hg), or prolonged macular detachment (more than 7 days), visual recovery in the primary vitrectomy group was significantly better (P <.05) than in the scleral buckle group from the first postoperative month. CONCLUSION: Both procedures achieved favorable anatomic and visual outcomes in the majority of patients with primary macula-off retinal detachment. Primary vitrectomy may be more effective than scleral buckling for achieving early visual rehabilitation in cases complicated by poor preoperative vision, ocular hypotony, and prolonged macular detachment.

Optical cross-sectional evaluation of successfully repaired idiopathic macular holes by retinal thickness analyzer.

PURPOSE: To describe optical cross-sectional images of successfully repaired idiopathic macular holes and to determine the influences of epiretinal membranes and retinal pigment epithelial (RPE) changes on foveal reconstruction and the correlation between retinal thickness and postoperative visual acuity. METHODS: In a prospective study, optical cross-sectional imaging and retinal thickness measurement of the macula using a retinal thickness analyzer were performed on 63 eyes of 63 patients who underwent successful macular hole surgery. RESULTS: Cross-sectional images of foveal reconstruction were morphologically categorized into four patterns: normal fovea (23 eyes [37%]), cavernous fovea (19 eyes [30%]), flat fovea (11 eyes [17%]), and irregular fovea (10 eyes [16%]). Epiretinal membranes were observed in the last two groups (55% and 40%) and RPE changes were observed only in the irregular fovea group (16%). The mean retinal thickness of the fovea in all eyes was 213 +/- 92 microm (mean +/- SD; range, 93-570 microm), which varied significantly (P < 0.001) among the four groups. Linear regression analysis showed a significant correlation between retinal thickness at the fovea and logarithmic converted visual acuity (R2 = 0.42, P < 0.001). CONCLUSIONS: Structural features of foveal reconstruction following successful macular hole surgery involved four patters: normal fovea, cavernous fovea, flat fovea, and irregular fovea. Retinal thickness of the fovea, which varied among the groups, correlated with postoperative visual acuity. Postoperative epiretinal membrane formation and RPE damage may disturb normal foveal reconstruction and visual recovery.

Phylogenic analysis of echovirus type 30 isolated from a large epidemic of aseptic meningitis in Japan during 1997-1998.

During 1997 to 1998, a nationwide epidemic of aseptic meningitis occurred in Japan. More than 4,500 isolates from patients with aseptic meningitis were identified as echovirus type 30. To investigate the character of these isolates, we examined the nucleotide sequences of thirty-seven geographical representatives and compared them with 50 strains isolated during the past 20 years. The phylogenic analysis used partial sequences from either the VP1 or VP4-VP2 region of the viral capsid. This analysis revealed that the isolates were divided into six genomic groups. All isolates identified during 1997-1998 belonged to only two genomic groups; these two groups are thought to be the causative viral agents involved in the recent epidemic.

[Macular morphologic reconstruction and visual recovery in patients following macular hole surgery].

PURPOSE: To examine the characteristics of retinal reconstruction at the macula following macular hole surgery and to assess the impact of postoperative macular morphological changes on the visual outcome. METHODS: A prototype of the scanning retinal thickness analyzer (RTA) was used to obtain optical section images at the macula in 53 eyes of 52 patients who underwent macular hole surgery between April 1994 and July 1997. The mean age of participants was 64.7 years. At the time of RTA examination, macular hole was biomicroscopically closed in 49 eyes (91%) and unclosed in 5 eyes (9%). RESULTS: There were 5 types of cross-sectional images of postoperative maculae: normal foveal depression [19 eyes (36%)], crater-like fovea formation [12 eyes (22%)], flattened fovea [8 eyes (15%)], fovea with abraded retinal pigment epithelium [9 eyes (17%)], and persistant macular hole [5 eyes (9%)]. Central macular thickness measured by RTA was 165 +/- 39 (mean +/- standard dereation) microns in the normal foveal depression group, 210 +/- 67 microns in the crater-like fovea formation group, and 300 +/- 50 microns in the flatened fovea group. There were statistically significant differences (p < 0.01) between groups. Postoperative visual acuity was significantly different (p < 0.01) between these three groups, and central macular thickness was highly correlated with postoperative visual acuity (R = 0.70). CONCLUSION: Visual recovery following macular hole surgery is closely associated with the retinal reconstruction at the macula.

Survey of surgical indications and results of primary pars plana vitrectomy for rhegmatogenous retinal detachments.

BACKGROUND: Several surgical techniques to repair rhegmatogenous retinal detachment have been developed. Recently, both the method of reattaching the retina and of obtaining an early visual recovery are considered important factors when determining which surgical techniques to perform to treat retinal detachment. CASES: The surgical outcome in a series of 63 consecutive patients, who were treated at Osaka Rosai Hospital between 1993 and 1996, was reviewed retrospectively to evaluate the efficacy of primary vitrectomy to treat uncomplicated rhegmatogenous retinal detachment associated with posterior hyaloid separation. The criteria for vitrectomy included the presence of not only posterior retinal breaks, but also of multiple peripheral retinal breaks. OBSERVATIONS: The reattachment rate after the first surgery was 92.1% (58 eyes), and by the final examination it increased to 100%. Of the 46 eyes with macular detachment, good visual rehabilitation and a visual acuity improvement of 5 or more lines was obtained in 33 eyes (71.7%) by 1 month postoperatively. No statistically significant difference in the reattachment rate was found when eyes that underwent an encircling procedure were compared with those that did not. In eyes with lens opacity, cataract surgery was also performed and intraocular lenses were implanted uneventfully in all but one case with myopia. There was a high incidence (53.8%) of cataract progression in phakic eyes. However, no other serious complications, such as proliferative vitreoretinopathy, were found throughout the follow-up period. CONCLUSIONS: The results indicate that vitrectomy performed to alleviate peripheral vitreoretinal traction is an effective surgical technique to treat primary rhegmatogenous retinal detachment. Vitrectomy combined with cataract surgery may also be a valuable surgical option in selected cases to maintain long-standing visual rehabilitation.

[A comparative study of visual outcomes following primary vitrectomy and scleral buckling procedures to manage macular off rhegmatogenous retinal detachments].

PURPOSE: To determine the difference between two types of retinal detachment surgery in postoperative visual recovery in the management of macular-off rhegmatogenous retinal detachment. METHODS: We evaluated retrospectively the postoperative visual acuity (VA) in 37 patients who underwent primary vitrectomy (group V) and 39 patients treated with conventional scleral buckling (group S). RESULTS: Although the mean preoperative VA in group V (0.03) was significantly worse (p = 0.04) than that in group S (0.06), there was no statistically significant difference between the groups in postoperative VA throughout the follow-up period. However, in the cases with poor preoperative VA (VA < 0.1), ocular hypotony (IOP < 7 mmHg), or prolonged macular detachment (duration > 7 days), the visual recovery was significantly better (p < 0.05) in group V than in group S from 1 month postoperatively. There were more eyes with a final VA of more than 0.5 in group V than in group S. When considering only the eyes with lenses spared intraoperatively, postoperative cataract progression resulting in secondary visual reduction was statistically significantly greater (p < 0.01) in group V (62%) than in group S (8%). CONCLUSIONS: Primary vitrectomy is effective to attain early visual rehabilitation, especially to manage macula-off retinal detachments with poor preoperative VA, ocular hypotony, and prolonged macular detachment. To prevent a secondary visual reduction, cataract surgery combined with vitrectomy is recommended in selected cases.

Quantitative assessment of macular thickness in normal subjects and patients with diabetic retinopathy by scanning retinal thickness analyser.

AIMS: To evaluate the scanning retinal thickness analyser (RTA), a novel non-invasive imaging instrument, in diagnosing and quantitatively characterising diabetic macular oedema, and to investigate the relation between central macula thickness measured by RTA and other clinical examinations. METHODS: Central macular thickness was measured using the RTA in 40 normal subjects and 60 patients with diabetic retinopathy. The reproducibility of the retinal thickness measurements was evaluated by calculating the mean of the inter- and intrasession variations. Central macular thickness was correlated with the results of visual acuity measurements, biomicroscopy, and fluorescein angiography. RESULTS: Intra- and intersession reproducibility of the RTA in normal subjects was plus or minus 5.2% (16 microns) and plus or minus 6.1% (19 microns), respectively. The mean central macular thickness was 182 (SD 16) microns in normal subjects, 283 (116) microns in diabetic eyes without clinically significant macular oedema (CSMO), and 564 (168) microns in diabetic eyes with CSMO. Central macular thickness was significantly greater (p < 0.001) in eyes with diabetic retinopathy than in normal subjects, even when macular thickening did not meet the standard for CSMO (p = 0.019) measured by biomicroscopy. Although greater fluorescein leakage at the macula results in greater central macular thickness, only eyes with diffuse leakage had statistically significant macular thickening compared with normal subjects (p = 0.022). Central macular thickness measured with the RTA was significantly correlated with the logarithmic converted visual acuity (r2 = 0.76) in diabetic eyes. CONCLUSION: Scanning RTA, which has good reproducibility, might be useful to quantitatively detect and monitor macular thickening in diabetic retinopathy. Central macular thickness was highly correlated with logarithmic converted visual acuity in diabetic macular oedema.

Serotypes of human rotaviruses in 7 regions of Japan from 1984 to 1997.

Human rotavirus (HRV) serotypes were studied from diarrheal stool specimens in children in 7 regions of Japan (Sapporo, Tokyo, Maizuru, Osaka, Kagawa, Kurume, and Saga) from 1984 to 1997 by enzyme immunoassay (EIA) with serotype-specific monoclonal antibodies against serotypes 1, 2, 3, and 4. In addition, reverse transcription-polymerase chain reaction (RT-PCR) was conducted for analysis of "others" which included nonserotypable and mixed-serotype rotavirus specimens by EIA. In 3756 rotavirus-positive specimens, serotype 1 was detected in 2649 (70.5%), serotype 2 in 362 (9.6%), serotype 3 in 232 (6.2%) and serotype 4 in 196 (5.2%). Overall, serotype 1 was predominant from 1984 to 1997, although there were a few cases in which serotype 2, 3 and 4 became predominant based on area and year. The frequency of serotype 1 has gradually increased since 1993. Twenty two, 2, 3 and 1 among 57 specimens of "others" by EIA from Tokyo, Maizuru, Sapporo and Kurume in 1995-1996 and 1996-1997 were determined as serotypes 1, 2, 3, and 9 by RT-PCR, respectively.

Long-term follow-up of primary anterior chamber intraocular lens implantation.

PURPOSE: To evaluate the long-term outcome in eyes having primary anterior chamber intraocular lens (AC IOL) implantation. SETTING: Kimura Eye and Internal Medicine Hospital, Hiroshima, Japan. METHODS: Primary AC IOL implantation was performed in 171 eyes from 1983 to 1988. Two lenses were implanted: the open C-loop Simcoe lens and the 4-point fixation Kelman lens. In 86 eyes that had a mean follow-up of 9 years 7 months, visual acuity, corneal endothelial cell density, pupil shape, and the presence of associated complications were examined. RESULTS: Although the final visual acuity was 0.5 or better in 60 eyes (69%), corneal endothelial cell density was less than 2000 cells/mm2 in 46 eyes (57%). Postoperative complications included a deformed pupil in 48 eyes (56%) and bullous keratopathy in 12 eyes (14%). In 14 eyes the AC IOL was explanted; 11 of these were the open C-loop Simcoe lens. CONCLUSION: Although our findings show that the open C-loop AC IOL (Simcoe) is contraindicated, implantation of the 4-point fixation AC IOL may be acceptable in selected cases.

Measuring the anterior capsule opening after cataract surgery to assess capsule shrinkage.

PURPOSE: To measure anterior capsule opening (ACO) size after acrylic intraocular lens (IOL) implantation and study the natural course of ACO reduction. SETTING: Kimura Eye and Internal Medicine Hospital, Hiroshima, Japan. METHODS: This study comprised 32 patients (38 eyes) having continuous curvilinear capsulorhexis, phacoemulsification, acrylic IOL implantation, and a self-sealing incision performed by 1 surgeon. A retroillumination photograph of the ACO was obtained with the Anterior Eye Segment Analysis System and converted to a computer image. The images were used to measure ACO size postoperatively and calculate the reduction ratio. Follow-up was 6 months. RESULTS: The postoperative reduction ratio in ACO size was 2.14% at 1 week, 3.83% at 1 month, 4.29% at 3 months, and 5.03% at 6 months. In a few cases, the reduction was progressively severe throughout the follow-up. CONCLUSIONS: The anterior capsule opening shrank rapidly during the first month after acrylic IOL implantation, followed by a slower progressive reduction in the subsequent 6 months. When severe, progressive shrinkage occurs, an anterior neodymium:YAG laser capsulotomy should be performed within 2 months postoperatively.

[Long-term results after simple vitrectomy for age-related macular degeneration with vitreous hemorrhage].

We reviewed the cases of 10 eyes with vitreous hemorrhage secondary to submacular hemorrhage associated with age-related macular degeneration, in which vitrectomy was performed without subretinal management. Vitreous hemorrhage occurred within 3 weeks of submacular hemorrhage. After vitrectomy, residual subretinal hemorrhage was rapidly absorbed and choroidal neovascularization was undetectable. After the surgery visual acuity improved over that of submacular hemorrhage in 5 eyes. Mean follow-up was 25 months and final visual acuity was 0.2 or better in 6 eyes (60%). Final visual outcome was highly corelated with the period from subretinal hemorrhage to vitrectomy, the thickness of subfoveal hemorrhage at vitrectomy, position of choroidal neovascularization, and duration of subretinal hemorrhage. Simple virectomy might be effective to stabilize submacular changes and improve visual acuity.

[Surgical indications and results of primary pars plana vitrectomy for rhegmatogenous retinal detachments].

To evaluate the efficasy of primary vitrectomy for uncomplicated rhegmatogenous retinal detachment associated with posterior hyaloid separation, a series of 63 eyes were reviewed retrospectively. The criteria for vitrectomy included not only eyes with posterior retinal breaks but also those with multiple peripheral retinal breaks. The reattachment rate was 92.1% (58 eyes) after the initial surgery, and finally this increased to 100%. Of the 46 eyes with macular detachment, good visual rehabilitation, i.e., visual acuity improved by 5 or more lines, was obtained in 71.8% (33 eyes) one month postoperatively. There was no statistically significant difference in the reattachment rate whether an encircling procedure was also done or not. In eyes with lens opacity, cataract surgery was also done and intraocular lenses were implanted uneventfully in all but one myopic case. No serious complications such as proliferative vitreoretinopathy were found throughout the follow-up period except for the high incidence (53.8%) of cataractous progression. The results indicate that vitrectomy, removing the peripheral vitreo-retinal traction directly, is an effective procedure for primary rhegmatogenous retinal detachment. Vitrectomy combined with cataract surgery is also proposed as a valuable strategy in selected cases to maintain visual rehabilitation.

Ligand efficacy and potency at recombinant alpha2 adrenergic receptors: agonist-mediated [35S]GTPgammaS binding.

Alpha-2 adrenergic receptors (alpha2 AR) mediate incorporation of guanosine 5'-O-(gamma-thio)triphosphate ([35S]GTPgammaS) into isolated membranes via receptor-catalyzed exchange of [35S]GTPgammaS for GDP. In the current study, we used [35S]GTPgammaS incorporation to characterize the intrinsic activity and potency of agonists and antagonists at the cloned mouse alpha2a/d and human alpha2a, alpha2b, and alpha2c ARs. Full agonists increased [35S]GTPgammaS binding to membranes by 2- to 3-fold. Antagonists did not increase [35S]GTPgammaS binding but competitively inhibited agonist-stimulated [35S]GTPgammaS binding. Compounds with intrinsic activities less than that of the full agonists norepinephrine (NE) or epinephrine (EPI) were capable of antagonizing agonist-stimulated [35S]GTPgammaS binding. The agonistic properties of a number of alpha2 AR ligands were characterized at each alpha2 AR subtype. The rank order of agonist potency for selected compounds at the human receptors (with intrinsic activity compared with NE, defined as 1.0) was: alpha2a: Dexmedetomidine (0.73) > guanabenz (0.38) > UK-14304 (1.02) > clonidine (0.32) > ST-91 (0.63) > NE (1.00). alpha2b: Dexmedetomidine (1.10) > clonidine (0.18) > guanabenz (0.71) > NE (1.00) > ST-91 (0.44) > UK-14304 (0.59). alpha2c: Dexmedetomidine (1.03) > NE (1.00) > UK-14304 (0.75) > ST-91 (0.32) > or = clonidine (0.23) >> guanabenz (0). This report provides a functional characterization of adrenergic receptor ligands at human and mouse alpha2a/d AR. It also illustrates the utility of [35S]GTPgammaS incorporation as a functional marker of receptor activation.

Molecular cloning, genomic characterization and expression of novel human alpha1A-adrenoceptor isoforms.

We have isolated and characterized from human prostate novel splice variants of the human alpha1A-adrenoceptor, several of which generate truncated products and one isoform, alpha(1A-4), which has the identical splice site as the three previously described isoforms. Long-PCR on human genomic DNA showed that the alpha(1A-4) exon is located between those encoding the alpha(1A-1) and alpha(1A-3) variants. CHO-K1 cells stably expressing alpha(1A-4) showed ligand binding properties similar to those of the other functional isoforms as well as agonist-stimulated inositol phosphate accumulation. Quantitative PCR analyses revealed that alpha(1A-4) is the most abundant isoform expressed in the prostate with high levels also detected in liver and heart.

Sequence analysis of SRSV in fecal specimens from an epidemic of infantile gastroenteritis, October to December 1995, Japan.

From October to December in 1995, an epidemic of infantile gastroenteritis occurred all over Japan except in Hokkaido and Okinawa prefectures. The number of infected infants and young children was estimated to be over 5 million cases [Editorial, IASR 1996]. The stool specimens from patients were examined for the presence of small round structured viruses (SRSVs) by reverse transcription-polymerase chain reaction (RT-PCR) and nucleotide sequencing of parts of the RNA-dependent RNA polymerase region. Thirty-five of 87 stool specimens examined gave positive results. Genomic variation was investigated by sequence analysis of a 327 bp cDNA region. The nucleotide and deduced amino acid sequences of the ten strains segregated into two distinct groups; one showed 96.0-100% nucleotide and 99.1-100% amino acid identity, the others showed 91.4-99.7% nucleotide and 93.5-100% identity. The main mechanism of transmission remains unknown. However, these data suggest the possibility of person-to-person spread by two or more kinds of SRSV.

Rheological evaluation of X-ray irradiated blood.

CPD blood, in glass containers and polyvinyl chloride (PVC) bags, was X-irradiated with 50 Gy (n = 4), and then stored for up to 4 weeks. Rheological, hematological and biochemical changes of the red cells during storage were examined. Changes in blood viscosity and various properties of red cells (deformability, shape, hematological indices and concentrations of 2,3-diphosphoglycerate and nucleotides) during storage at 4 degrees C were not altered by the irradiation. The rheological, biochemical and morphological properties of red cells were as well preserved in PVC bags as in glass containers. No abnormal and/or cross-linked proteins in red-cell membranes were induced by the irradiation. Irradiation of PVC bags did not affect the above properties of red cells through any changes in the bag constituents. In conclusion, X-ray irradiation with 50 Gy to CPD blood in PVC bags does not affect the rheological functions of red cells.

Multifrequency laser emission generated by two-color stimulated Raman effect using a single-frequency laser beam and a dye-Raman composite resonator.

A single-color circularly polarized laser beam is focused into pressurized hydrogen to generate rotational stimulated Raman emission. After being corrected to a linearly polarized beam, this two-color laser beam is passed through a dye amplifier and is focused into pressurized hydrogen, again to generate multifrequency laser emission by the two-color stimulated Raman effect. The laser beam is further confined in a resonator, which enhances intracavity intensity leading to increased efficiency. In these experiments, more than 11 rotational lines are generated simultaneously at 5 atm, and many additional vibrationally shifted rotational lines are generated at 9 atm.