TT virus mRNAs detected in the bone marrow cells from an infected individual.

Although replicative forms of TT virus (TTV) DNA have been found in the liver and bone marrow cells, mRNAs of TTV have not yet been detected in these tissues. The entire nucleotide sequence of a TTV clone [TYM9 (3759 bases)] isolated from a patient with high TTV viremia (10(6) copies/ml) was determined, and the poly(A)(+) RNAs from bone marrow cells were subjected to reverse transcription-polymerase chain reaction with primers specific for the TYM9 sequence. Sequence analysis of the amplified products revealed the presence of three distinct species of spliced TTV mRNAs [2.9, 1.2, and 1.0 kilobases (kb)] with common 5' and 3' termini as well as splicing to bind nucleotide (nt) 181 to nt 283. The shorter mRNAs of 1.2 kb and 1.0 kb possessed another splicing to join nt 681 with nt 2341 or nt 2579. The transcription profile of TTV found in an infected human corroborates that observed in vitro.

Sequestration of TT virus of restricted genotypes in peripheral blood mononuclear cells.

Peripheral blood mononuclear cells (PBMC) harbored TT virus (TTV) of genotypes (3 and 4) different from those (1 and 2) of free virions in plasma of the same individuals. PBMC may act as a reservoir, and TTV of particular genotypes might have tropism for hematopoietic cells.

Immunohistochemical localization of MYOC/TIGR protein in the trabecular tissue of normal and glaucomatous eyes.

PURPOSE: To examine immunohistochemically the localization of myocilin/trabecular meshwork inducible glucocorticoid response (MYOC/TIGR) protein in the glaucomatous and normal trabecular meshworks. METHODS: Trabecular tissues were used from one eye with late-onset goniodysgenetic glaucoma, three with primary open angle glaucoma (one of which had the MYOC/TIGR gene mutation), two with exfoliation glaucoma and one without glaucoma. For light microscopic immunohistochemistry, frozen sections were stained by the avidin-biotin complex method using anti-MYOC/TIGR polyclonal antibody. For electron microscopic immunohistochemistry, the pre-embedding method using the same antibody was performed. Double immunostaining using both anti-MYOC/TIGR and anti-type VI collagen antibodies was done by the immunofluorescence method. RESULTS: With light microscopy, immunoreactivity was seen in the whole trabecular meshwork of each of the specimens. No notable differences were detected in staining among the types of glaucoma, or between the eyes with and those without the gene mutation. Under electron microscopy, immunoreaction products were observed not only in the cytoplasm of the trabecular cells but also in the extracellular matrix, where staining was associated with the long-spacing collagen, fine granular materials and possibly microfibrils. With double immunohistochemistry, MYOC/TIGR was colocalized with type VI collagen in the trabecular meshwork. CONCLUSIONS: In glaucomatous and normal trabecular meshworks, the MYOC/TIGR protein is distributed in the extracellular matrix colocalizing with type VI collagen.

Replicative forms of TT virus DNA in bone marrow cells.

TT virus (TTV) is a human virus consisting of a single-stranded, circular DNA genome of 3.8 kilobases (kb). To examine whether TTV replicates in peripheral blood mononuclear cells (PBMCs) and bone marrow cells (BMCs), DNA was extracted from the PBMCs and/or BMCs of six TTV-infected individuals and separated by agarose gel electrophoresis. The TTV DNAs from the PBMCs migrated to the 2.0- to 2.5-kb region. The TTV DNAs from the BMCs migrated to the 2.0- to 2. 5-kb and 3.3- to 6.1-kb regions. The faster-migrating TTV DNAs were sensitive to S1 nuclease, while the slower-migrating TTV DNAs were resistant and their position on the agarose gel shifted to the position of the full genomic size upon digestion with restriction enzyme PstI. Full-length inverted polymerase chain reaction on the slower-migrating, double-stranded TTV DNAs from the BMCs amplified a 3.8-kb product. Replicative intermediate forms of TTV DNA are present in BMCs but not in PBMCs.

Postmortem findings two weeks after oral treatment for metastatic Candida endophthalmitis with fluconazole.

The purpose of this histological study was to present postmortem findings in both eyes of a 53-year-old male with liver dysfunction 2 weeks after short-time oral treatment with 200 mg/day fluconazole for metastatic Candida endophthalmitis due to intravenous hyperalimentation for 18 days. Candida had been demonstrated in the venous blood and on the tip of the intravenous catheter. The bilateral fungal endophthalmitis with hypopyon responded well to fungistatic therapy, but the patient suddenly died from heart failure. Both eyes were obtained at autopsy. Candida was demonstrated only in vitreous puff balls but not in the retina or uvea. Fluconazole administered for a short period had little effect in eliminating fungus from vitreous puff balls, which have no blood supply. Prolonged administration of the antifungal drug or vitrectomy should be considered when treating an eye with vitreous puff balls in the presence of fungal endophthalmitis.

[Iris vessel involvement in iris-nevus syndrome].

PURPOSE: To report a case of iris-nevus syndrome accompanied by disruption of the blood-aqueous barrier in the iris which was confirmed angiographically and histopathologically. CASE: The patient was a 39-year-old woman. She noticed blurred vision in the left eye which was diagnosed as left glaucoma. Specular microscopy revealed low endothelial cell density in the left cornea. The left iris showed atrophy with clusters of nodular iris nevus and distorted pupil. The left iridocorneal angle was closed with peripheral anterior synechia. FINDINGS: Indocyanine green iris angiography revealed more vessels on the surface of the left iris than on the right. In fluorescein iris angiography, the dye leaked from the iris vessels in areas where the iris showed advanced atrophy. The trabecular tissue obtained by trabeculectomy from the patient's left eye showed histopathologically a lining of corneal endothelial cells on the surface of the iris. The density of the vessels was high in the iris stroma. Some cells covering the vessel wall showed degeneration with opening of the zonula occludens. Schlemm's canal had narrowed lumina, and the intertrabecular spaces were closed. CONCLUSION: Disruption of the blood-aqueous barrier may occur in iris-nevus syndrome.

Ciliary body band width as an indicator of goniodysgenesis.

PURPOSE: To evaluate the gonioscopic signs indicative of underdevelopment of the iridocorneal angle and thereby obtain a correct diagnosis of goniodysgenetic glaucoma. METHODS: We examined morphologically the iridocorneal angle of 28 enucleated human eyes, including those from premature infants as well as from older subjects. The ratio of the ciliary body band width to the width of the posterior limbus (the distance between the termination of Descemet's membrane and the iris root), termed the theoretic C/L ratio, was calculated by drawing on light micrographs. Data were compared for three age groups: premature infants, children, and subjects older than 9 years. We also examined clinically the appearance of the ciliary body band on gonioscopy in 135 eyes of 69 younger patients having primary glaucoma with open iridocorneal angles and in 94 eyes of 47 age-matched control subjects. RESULTS: The theoretic C/L ratio +/-SD was 1.4% +/- 2.7% in premature infants, 24.0% +/- 5.4% in children, and 36.7% +/- 8.0% in the older subjects. There was a statistically significant difference between each of the three groups (between premature infants and children, P < .001; between children and older subjects, P < .005; between premature infants and older subjects, P < .001). On gonioscopy, 105 of 135 glaucomatous eyes (77.8%) and 13 of 94 control eyes (13.8%) showed either an invisible or very narrow ciliary body band (P < .0001). CONCLUSION: The ciliary body band seen on gonioscopy provides an indicator of the development of the iridocorneal angle. An invisible or very narrow ciliary body band represents an underdevelopment of the angle.

Neovascular tissue in the intertrabecular spaces in eyes with neovascular glaucoma.

AIMS/BACKGROUND: To investigate histological changes in the trabecular meshwork in eyes with neovascular glaucoma. METHODS: Light and electron microscopic studies were carried out on the trabecular meshwork of three enucleated eyes with neovascular glaucoma. The presence and distribution of factor VIII in the trabecular meshwork was assessed using the ABC method. RESULTS: Peripheral anterior synechiae covering the trabecular meshwork were detected in two eyes, which would explain the rise in intraocular pressure. In the third the angle was not completely closed by peripheral anterior synechiae. The spaces between the trabecular beams were lined by a single layer of vascular endothelium, and were filled with red blood cells in this patient. Factor VIII was positively stained in the endothelial cells, lining both these spaces and Schlemm's canal. A basal lamina and microfibrils were detected just beneath the newly formed vascular endothelial cells. CONCLUSION: The neovascular tissue found in the trabecular spaces might be one of the factors responsible for intraocular pressure elevation in eyes with neovascular glaucoma.

Anterior chamber angle vascularization in Sturge-Weber syndrome. Report of a case.

The case of a 20-year-old woman with a left-sided facial hemangioma and a homolateral glaucoma is reported, complete with the histology of a trabeculectomy specimen. Her left eye had an episcleral hemangioma and goniodysgenetic features in the anterior chamber angle, while the intraocular pressure was measured to be 45 mmHg. The left optic disc showed a large cupping and the left visual field was constricted. The right eye had no glaucomatous changes. Histological examination of the trabeculectomy specimen by both light and electron microscopy showed multiple congenital anomalies. There was a cluster of blood vessels in the trabecular meshwork. Abnormal accumulations of fine granular extracellular matrixes were observed in both the juxtacanalicular connective tissue and around the vascular structures. The lumen of Schlemm's canal was subdivided into three or four parts with few giant vacuole structures. The endothelial cells lining the inner wall of Schlemm's canal contained a well-formed basal lamina with many villi projecting into the lumen. These findings suggest that the multiple anomalies observed in the trabecular tissue may contribute to the manifestation of glaucoma in Sturge-Weber syndrome.

Histological study on transient ocular hypertension after laser iridotomy in rabbits.

We performed argon-laser iridotomy on pigmented rabbits and examined histologically the tissue of the anterior chamber angle, the iris, and the ciliary body. After laser iridotomy, 9/10 eyes showed an elevation in transient intraocular pressure. Deposits of blood plasma with fibrin were observed in the tissue of the circumferential anterior chamber angle from eyes enucleated at a period of high intraocular pressure. In the eyes in which ocular pressures returned to the baseline, the quantity of blood plasma was small. In the laser-irradiated iris and iris processes, cells constructing the vessel wall were degenerated, and blood plasma and fibrin exuded. In and around the ciliary body, fibrin was not demonstrated. These observations suggest that argon-laser irradiation to the iris leads to a breakdown of the blood-aqueous barrier mainly in the laser-treated iris, and that deposition of blood plasma with fibrin in the anterior chamber angle tissue may cause transient intraocular pressure elevations after laser iridotomy in rabbits.

Transformation and cytotoxicity of iron in siderosis bulbi.

Ocular tissues from two enucleated and one trabeculectomized human eyes with siderosis bulbi were studied by electron microscopy. Ferritin particles were demonstrated in various cell types of the three eyes. Other types of iron were not identifiable. The ferritin particles were scattered throughout the cytoplasm, with a few particles in the nucleus and extracellular space. More ferritin particles were seen in tissues near the iron foreign body. The cells which contained numerous ferritin particles also had siderosomes, most of which were thought to be conglomerates of the ferritin particles in the secondary lysosomes. Cells with numerous ferritin particles, particularly in the siderosomes, showed vacuolar degeneration. The present study indicated that in siderosis bulbi, iron released from the iron foreign body is deposited as ferritin scattered throughout the cytoplasm and is sometimes accumulated as siderosomes, in the affinitive cells. In siderosis bulbi, the cells become damaged by the deposition of ferritin in the cytoplasm, especially in the form of siderosomes.

Anterior and posterior parts of human trabecular meshwork.

From gonioscopic observations, the trabecular meshwork was tentatively divided into anterior and posterior parts. The anterior part was defined as the zone from the Schwalbe's line to the anterior edge of the Schlemm's canal, and the posterior part was from the anterior edge of the Schlemm's canal to the angle recess. The patients' own blood cells were injected into the anterior chamber of two eyes about to be enucleated due to malignant tumors, and cell distribution in the trabecular meshwork was examined histologically. Blood cells in the posterior part of the trabecular meshwork were numerous, particularly in the area adjacent to the Schlemm's canal and anterior to the ciliary muscle at the angle recess, yet only a few cells were seen in the anterior part. At the angle recess, the blood cells accumulated in the perivascular connective tissue of the major arterial circle of the iris. These observations suggest that the posterior part of the trabecular meshwork is the most important for aqueous humor drainage in the human eye, both with regard to the conventional and unconventional routes, while the anterior part plays a lesser role in the aqueous outflow. It is also suggested that some proportion of the aqueous humor entering the tissues at the angle recess may flow through the perivascular tissue of the major arterial circle of the iris.

Case of sarcoidosis with uncommon tumorous mass formation in bulbar conjunctiva.

A case of sarcoidosis with ocular involvement, including uncommon tumorous mass formation on the bulbar conjunctiva, was reported. The patient was a 62-year-old woman who had been suffering from chronic bronchiolitis for several years. The conjuctival masses, yellowish brown in color with localized injection and follicles, were found in both eyes. In addition to the conjunctival masses, the common manifestations of sarcoidosis were observed in the eyes, showing nodules on the iris and trabeculum, discrete fluffy "snowball" opacities in the vitreous, and perivascular nodules on the retinal vessels. Microscopically, the biopsy specimen obtained from he conjunctival masses was composed of epithelioid tubercles with Langhans type and foreign body type giant cells. Ziehl-Neelsen stain revealed no tuberculous bacilli. Transbronchial lung biopsy also revealed epithelioid tubercles. Both ocular and pulmonary lesions were observed to respond well to treatment with corticosteroid hormone. We emphasized that patients with interstitial pneumonitis or pulmonary fibrosis should be examined ophthalmologically, and recommended conjunctival biopsy in patients with suspected sarcoidosis.