Fibrocartilaginous embolism in a chondrodystrophoid breed dog.

An 8-year-old intact male Shih Tzu dog was admitted with acute-onset tetraplegia. Magnetic resonance imaging showed an abnormality of the cervical intramedullary spinal cord. Histopathological examination of a spinal biopsy confirmed the presence of intravascular cartilaginous emboli. This is the first report of an antemortem diagnosis of fibrocartilaginous embolism in a chondrodystrophoid breed dog.

D-penicillamine-induced pemphigus successfully treated with combination therapy of mizoribine and prednisolone.

A 63-year-old woman had D-penicillamine-induced pemphigus with a high index value of circulating autoantibodies against desmoglein 1. Because of the lack of response to prednisolone monotherapy, the patient was treated with a combination therapy of mizoribine and low-dose prednisolone. The skin eruption was improved by this combination with a declined value of circulating anti-desmoglein 1 antibody. Currently, a remission of 2 months has been achieved, and prednisolone could be tapered.

[Immunohistochemical study of idiopathic and secondary epiretinal membrane].

PURPOSE: We investigated the immunohistochemical features of surgically resected idiopathic epiretinal membranes(ERMs) and secondary ERMs with regard to posterior vitreous detachment(PVD). METHODS: Six specimens of idiopathic epiretinal membranes(3 eyes with complete PVD, 2 eyes with partial PVD, and one eye with no PVD) and 3 specimens of secondary ERMs(all eyes with complete PVD) were immunohistochemically studied. We used type I, II, III, IV collagen and fibronectin to study extracellular components, and glial fibrillary acidic protein(GFAP), S 100 protein, vimentin, and so forth to study cellular components. RESULTS: All the specimens of idiopathic ERMs had the major components of the lamellar stained by type II collagen antibody, and one out of 3 specimens of secondary ERMs had a minor component stained by type II collagen antibody. Compared with idiopathic ERMs with complete PVD, 2 out of 3 specimens of idiopathic ERMs with partial PVD or no PVD contained rather thick collagen lamellar. CONCLUSION: There was difference between specimens of idiopathic ERMs and specimens of secondary ERMs in staining by type II collagen antibody, supposed by vitreous, in this study. Idiopathic ERM with attached posterior vitreous membrane may cause growth of collagen.

Proteus syndrome.

A case of Proteus syndrome is presented, in which severe hemihypertrophy of the left trunk and left lower extremity, scoliosis, endometriosis and huge bizarre-shaped body tumors were observed. Up to 22.6 kg of tumorous tissue was excised. This syndrome was first described in 1983. The name Proteus comes from a Greek mythical sea god who was able to change his body form freely. This syndrome has numerous features including hemihypertrophy, macrodactyly, various subcutaneous masses, scoliosis and other minor abnormalities. Although diagnostic criteria have been established for Proteus syndrome, which is very difficult to differentiate from other congenital hamartomatous syndromes, more case reports are needed to define such a rare disorder. Our patient is the 6th Japanese case in the English literature.

Two novel glucose-6-phosphate dehydrogenase variants found in newborn mass-screening for galactosaemia.

UNLABELLED: Glucose-6-phosphate dehydrogenase (G6PD) deficiency is an X-linked recessive disorder in which haemolytic anaemia is the major symptom. The Beutler spot test employed in mass-screening for galactosaemia in newborns requires several intrinsic erythrocyte enzymes such as G6PD for its reaction and can theoretically detect G6PD deficiency apart from galactose-1-phosphate uridyltransferase deficiency. In this study, we detected two patients with G6PD deficiency using the quantitative Beutler test which was recently developed in our laboratory. Both patients lacked erythrocyte G6PD activity but exhibited no clinical symptoms. Molecular analysis in patients 1 and 2 revealed two novel missense mutations of C853T causing R285C and A1220C causing K407T, respectively. Molecular rather than enzymatic analysis was required in familial studies to detect and diagnose the carrier state. To date these patients have avoided oxidant stress and haemolytic diatheses have not been induced. CONCLUSION: Our results indicate that the quantitative Beutler test can detect glucose-6-phosphate dehydrogenase deficiency of class 1 and 2 and is therefore useful for early intervention and prevention of haemolytic diathesis in patients with this disorder.

Analysis of radiation damage of DNA by atomic force microscopy in comparison with agarose gel electrophoresis studies.

DNA damage induced with ionizing radiation is considered one of the main causes of cell inactivation. Several methods including gel electrophoresis, pulsed-field gel electrophoresis, neutral filter elution method, neutral sedimentation and electron microscopy have been applied to analyze this type of DNA damage. A new method employing an atomic force microscope (AFM) for nanometer-level-structure analysis of DNA damage induced with gamma-irradiation is introduced in this report. Structural changes of plasmid DNA on a molecular size scale of about 3 kbp were visually analyzed by AFM after irradiation with 60Co gamma-rays at doses of 1.9, 5.6, and 8.3 kGy. Three forms of plasmid DNA, closed circular (intact DNA), open circular (DNA with a single strand break) and linear form (DNA with a double strand break) were visualized by dynamic force mode AFM after gamma-irradiation. The torsional feature of the plasmid DNA was visualized better with AFM than with a transmission electron microscope (TEM). All three forms of plasmid DNA were observed in the sample irradiated with gamma-rays at the dose of 1.9 kGy. Open circular and linear forms were observed in the samples irradiated with gamma-rays at doses of 5.6 and 8.3 kGy, though no closed circular form was observed. A shortening of the length of a linear form of DNA irradiated with 5.6 and 8.3 kGy gamma-rays was observed by AFM. Structural changes of DNA after gamma-irradiation were visualized by AFM at nanometer level resolution. In addition, shortening of the length of the linear form of DNA after radiation exposure was observed by AFM.

Acute relapsing encephalopathy mimicking acute necrotizing encephalopathy in a 4-year-old boy.

A 4-year-old boy showed two episodes of encephalitis/encephalopathy involving disturbed consciousness, convulsion, and paresis associated with the elevated levels of protein and myelin basic protein of the cerebrospinal fluid. MRI studies of the brain revealed symmetrical lesions in the brain stem and thalami at the first episode, and additional lesions were found in the cerebellum involving both the gray and white matter in the second episode. The intensities of MRI lesions were low in T I and high in T2. These episodes were followed by an elevation of the anti-viral antibody titers, for influenza A virus during the first episode and for adenovirus during the second. In the second episode, intravenous methylprednisolone therapy resulted in rapid improvement of his neurological signs.

Molecular basis for phenotypic heterogeneity in galactosaemia: prediction of clinical phenotype from genotype in Japanese patients.

We identified 14 mutations in 15 Japanese subjects from 13 families with galactose-1-phosphate uridyltransferase (GALT) deficiency using denaturing gradient gel electrophoresis (DGGE) and direct sequence analysis. These mutations accounted for 22 (96%) of 23 mutant alleles in 15 Japanese subjects. The mutational spectrum included nine missense mutations (M142V, G179D, A199T, R231H, W249R, N314D, P325L, R333Q, and R333W), two deletions (L275fsdelT and Q317fsdelC), a nonsense mutation (W249X), and two splicing mutations (V85-N97fsdel38bp and IVS4nt+1). Ten of the 14 mutations have not been reported in Caucasians. Differences in frequency and spectrum of GALT mutations suggest that the mutations may have occurred after racial divergence of Caucasians and Asians. The Duarte variant in Japanese was associated with the N314D mutation, g.1105G > C, g.1323G > A, and g.1391G > A (SacI -) polymorphisms, as in Caucasians. The Duarte variant may have occurred before racial divergence, and was an ancient mutation. In vitro GALT activities of nine missense mutations were determined by a COS cell expression system, and indicated between 1.3% and 35% of wild-type control. Patients with R333Q (29% in vitro GALT activity) or A199T (35%) showed mild clinical phenotypes, i.e. no ovarian failure or neurological deterioration. Genotype determination is useful for predicting biochemical and clinical phenotypes in classic galactosaemia, and can be of further help in managing patients with this disorder.

[Telemedicine and telescience in the field of ophthalmology at Asahikawa Medical College].

Department of Ophthalmology at Asahikawa Medical University has established a video conferencing telemedicine network system between the department and each branch hospital in Japan to transmit full-motion (30 frames/second) color ophthalmological images since October 1994. Nowadays, the department has 15 branches to communicate with our system including one hospital in Fukushima Prefecture; Department of Ophthalmology, Harvard Medical School in USA; Nanjing Medical University in China with the use of INS net 1500 or INS net 64 (3 lines). This system is aimed for telemedicine, telescience as well as supports for ophthalmological surgeries. In this article, we described the present status and future prospective of our telemedicine project for the purpose of the elimination of areas without enough medical facilities and promotion of international scientific collaboration. We also described our "Telemedicine Center" at Asahikawa Medical University which is now under construction. We believe that this technology literally opens up the world to scientific and medical collaboration.

Oral fluorescein angiography with the confocal scanning laser ophthalmoscope.

OBJECTIVE: To evaluate the efficacy of oral fluorescein angiography with a confocal scanning laser ophthalmoscope (SLO) system. DESIGN: Comparative case series. PARTICIPANTS: The authors used a confocal SLO (Heidelberg Retina Angiograph [HRA]) to perform oral fluorescein angiography in 47 patients, 13 of whom were without any retinal disease and 34 with a variety of retinal diseases including macular holes and pucker, inflammatory diseases, retinal vascular diseases, and age-related macular degeneration. The images were also compared to images taken with a fundus camera after intravenous fluorescein injections in patients on whom both studies were done. INTERVENTION: Color fundus photographs were taken of each eye (30 degrees fundus camera) before drinking 4 ml of 25% sodium fluorescein mixed with 60 ml of orange juice. After oral fluorescein ingestion, images of each eye were taken with a fundus camera (TriX film) and the HRA (using 512- x 512-pixel resolution). The images were repeated at 0-, 2.5-, 5-, 7.5-, 10-, 12.5-, 15-, 20-, 25-, and 30-minute intervals. Twenty of the 47 patients underwent intravenous fluorescein angiography performed with the fundus camera. MAIN OUTCOME MEASURE: Images were analyzed by a masked reader, and foveal avascular zone visualization, branch retinal vessel identification, and image quality were scored. Statistical analysis was performed with a t test for paired data with a two-tailed test of significance (alpha = 0.05). RESULTS: Foveal avascular zone was 100% as seen in 16 eyes (47%) in the HRA machine versus 1 eye (2%) in the conventional fundus camera (P < 0.0001). The third-order branch retinal vessels were identified in 59% of eyes in the HRA versus 26% in the fundus camera group (P < 0.0001), and the image quality was considered comparable to an intravenous angiogram in 47% with the HRA versus 9% with the conventional fundus camera (P < 0.0001). CONCLUSIONS: Oral fluorescein angiography using the HRA produces sufficiently detailed images to diagnose, treat, and follow many types of retinal pathology.

Gene 1, one of the dna genes of bacteriophage phi 29, represses other dna genes through binding to mRNA.

The dna genes, essential for protein priming DNA replication of bacteriophage phi 29, are transcribed as a long polycistronic mRNA. In the previous study, gene 1 product (gp1) was shown to repress the expression of the upstream dna genes for DNA polymerase and primer protein. To investigate the details of the repression by gp1, we have examined the amount and integrity of polycistronic mRNA encoding DNA polymerase and primer protein by agarose gel electrophoresis and nuclease S1 protection assay. As a result, the amount, size, and integrity of the polycistronic mRNA were not influenced by the presence of gene 1. Furthermore, the RNA binding ability of gp1 was demonstrated by in vitro system using histidine-tagged gp1. These results strongly suggested that translation of the dna genes was affected by gp1 through binding to mRNA. Other possible mechanisms of gene regulation by gp1 were discussed.

Comparison of short- and long-term effects of betaxolol and timolol on human retinal circulation.

PURPOSE: To determine the short- and long-term effects of betaxolol and timolol on human retinal circulation. METHODS: In a double-masked, randomised, placebo-controlled study we evaluated the effects of both a one-drop application and a twice-daily 2-week application of either topical 0.5% betaxolol hydrochloride or topical 0.5% timolol maleate on the retinal circulation in 12 healthy volunteers. Laser Doppler velocimetry was used to detect changes in the retinal venous blood flow. RESULTS: In both betaxolol- and timolol-treated eyes, intraocular pressure decreased significantly compared with baseline values after both 90 min and 2 weeks. In betaxolol-treated eyes, retinal blood flow did not change significantly after 90 min, but increased significantly (14 +/- 9%; p = 0.02) compared with baseline after 2 weeks. In timolol-treated eyes, retinal blood flow decreased significantly (18 +/- 5%: p = 0.04) compared with baseline after 90 min, and also decreased significantly (14 +/- 6%; p = 0.04) compared with baseline after 2 weeks. CONCLUSIONS: Retinal blood flow increases as a long-term effect of betaxolol and decreases as both a short- and long-term effects of timolol.

Abnormal vitreous structure in optic nerve pit.

A 37-year-old man presented with an optic nerve pit and serous macular detachment of the left eye. Scanning laser ophthalmoscopy revealed a cyst-like structure terminating at the pit in the premacular vitreous. During ocular movement, this structure moved vigorously and seemed to exert traction on the pit. We believe that it is part of an anomalous Cloquet's canal, and that traction on the pit may be a significant factor in the development of serous macular detachment in this patient.

[Cousins with X-linked recessive myotubular myopathy].

We describe two cousins with severe infantile form of myotubular myopathy. In Japan this disease has previously been reported in only three families. Case 1. The propositus, a 2-year-5-month-old boy, had been on a respirator since birth. He had a history of severe neonatal asphyxia and sequential hypotonia with dyspnea. Findings diagnostic of congenital myotubular myopathy, such as central nuclei and peripheral halo of muscle fibers, were demonstrated in his biopsied muscle. Case 2. A male the cousin of case 1 had congenital myopathy and died at 3 months of age due to respiratory failure. His muscle biopsy disclosed the identical findings as had been seen in case 1. These two cases were born to twin mothers, suggesting X-linked recessive inheritance. Early diagnosis and proper treatment of myotubular myopathy are important, because this condition may be erroneously-interpreted as the sequelae of neonatal asphyxia.

Variations of posterior vitreous detachment.

AIMS: To identify variations in posterior vitreous detachment (PVD) and establish a clinical classification system for PVD. METHODS: 400 consecutive eyes were examined using biomicroscopy and vitreous photography and classified the PVD variations-complete PVD with collapse, complete PVD without collapse, partial PVD with thickened posterior vitreous cortex (TPVC), or partial PVD without TPVC. RESULTS: In each PVD type, the most frequently seen ocular pathologies were as follows: in complete PVD with collapse (186 eyes), age related changes without vitreoretinal diseases (77 eyes, 41.4%) and high myopia (55 eyes, 29.6%); in complete PVD without collapse (39 eyes), uveitis (23 eyes, 59.0%) and central retinal vein occlusions (8 eyes, 20.5%); in partial PVD with TPVC (64 eyes), proliferative diabetic retinopathy (30 eyes, 46.9%); and inpartial PVD without TPVC (111 eyes), age related changes without vitreoretinal diseases (62 eyes, 55.9%). This PVD categorisation was significantly associated with the prevalence of each vitreoretinal disease (p < 0.0001, chi 2 test on contingency table). CONCLUSIONS: PVD variations can be classified into four types, which is clinically useful because each type corresponds well to specific vitreoretinal changes.

Electron microscopy of biological specimens by the plasma-polymerization rapid-freeze replica method.

The plasma-polymerization replica method is a unique replica technique for transmission electron microscopy. In the present study, we used this method in combination with a rapid-freeze technique to observe T4 bacteriophages and hepatitis B virus core particles. The heads of T4 bacteriophages appeared hexagonal and measured approximately 110 nm in length. Striations in their tails were also visible, indicating that the resolution of the present method is better than 4 nm. The images corresponded well with those obtained by ice-embedding and negative staining methods, with respect to both morphology and size of the phage particle. Hepatitis B virus core particles observed by the present method appeared round, approximately 30 nm in diameter, with hollow centres. Again, the morphology and size of the particles corresponded well with those obtained by ice-embedding, negative staining, and ultrathin sectioning. From these results, we conclude that the plasma-polymerization rapid-freeze replica method provides a useful technique for observation of biological specimens in a natural state and at high resolution.

[Biomicroscopic findings of premacular posterior vitreous].

The presence of vitreous gel in front of the macular area is still controversial. In order to understand the anatomy of the premacular vitreous, the posterior vitreous was observed biomicroscopically and slit-lamp photographs were taken in 100 eyes without posterior vitreous detachment. We defined the premacular vitreous as the lacuna seen when optically empty space, a demarcated oval shaped dark area without Tyndall phenomenon, was observed in front of the macula. Although the premacular vitreous showed liquefaction, the Tyndall-phenomenon, indicating the presence of formed vitreous gel, was observed in most cases. Premacular lacuna was observed in 3 cases with high myopia, and in one case with vitreoretinal degeneration syndrome. The bursa premacularis or premacular precortical vitreous pocket observed in autopsy eyes could barely be observed in living eyes.

Comparison of the prevalence of posterior vitreous detachment in whites and Japanese.

The vitreous condition of healthy white (n = 551) and Japanese (n = 528) eyes were reviewed and compared with respect to the prevalence of posterior vitreous detachment (PVD) in the two groups. No white or Japanese patient 39 years of age or younger had PVD; the prevalence among those older than 39 increased with age in the fifth through the ninth decades, i.e., respectively, to 4%, 24%, 37% 59%, and 87% in the whites, and to 5%, 21%, 43%, 72%, and 82% in the Japanese. In none of these decades was there any significant difference between the prevalence of complete or partial PVD in the whites and the Japanese. This finding is significant because the vitreoretinal relationship influences the development and prognoses of the various vitreoretinal disorders.