Screening for mutations of Axenfeld-Rieger syndrome caused by FOXC1 gene in Japanese patients.

PURPOSE: Mutations in the forkhead transcription factor gene (FOXC1) have been recently shown to cause some cases of juvenile glaucoma associated with a variety of anterior-segment anomalies. The purpose of this study was to investigate the clinical features of Axenfeld-Rieger syndrome caused by FOXC1 mutations in Japanese patients. PATIENTS AND METHODS: After informed consent was obtained, genomic DNA was isolated from peripheral blood. The DNA-sequence changes were analyzed using single-strand conformation polymorphism analysis and automated sequencing in six Japanese probands with Axenfeld-Rieger syndrome. RESULTS: The authors identified four mutations: pedigree 1 (26-47ins22), 2 (Ile91Ser), 3 (286ins1), and 4 (Arg127His). Two pedigrees showed new mutations in FOXC1. In pedigrees 1,2, and 4, younger generations had iris hypoplasia with severe early-onset glaucoma, whereas their parents had posterior embryotoxon without glaucoma. Pedigree 3 had a single affected person with iris hypoplasia and posterior embryotoxon with a mild increase of intraocular pressure. CONCLUSION: Four different FOXC1 mutations were found in four of six Japanese pedigrees with Axenfeld-Rieger syndrome. This was a new mutation in two pedigrees that was not found in earlier generations. This study confirms that mutations in this gene cause maldevelopment of the anterior segment of the eye.

[Neuroprotective effect of N-methyl-D-aspartate receptor antagonists in an experimental glaucoma model in the rat].

PURPOSE: To evaluate the neuroprotective effect of memantine and dizocilpine, which are noncompetitive open-channel blockers of the N-methyl-D-aspartate (NMDA) receptor, on glaucomatous optic neuropathy in an experimental glaucoma model in the rat. METHODS: Experimental glaucoma was induced in the right eyes of 30 Wistar albino rats by intracameral injection of India ink followed by laser trabecular photocoagulation 4 days later. The left eye served as a control. Either memantine, dizocilpine, or phosphate-buffered saline (PBS) was injected intraperitoneally just before trabecular photocoagulation. Five days later, 3% fast blue was injected into both superior colliculi. The eyes were enucleated another 3 days later and flat mounts of the retinas were prepared. Labeled ganglion cells were counted in the area 1 mm away from the optic disc. RESULTS: Five days after laser application, no significant intraocular pressure (IOP) change in the right eye was found among the 3 groups. In eyes treated with memantine or dizocilpine, significantly more ganglion cells were labeled. CONCLUSION: Systemically applied memantine and dizocilpine had a neuroprotective effect against experimental glaucomatous optic neuropathy in the rat.

Optic nerve and peripapillary choroidal microvasculature of the rat eye.

PURPOSE: To investigate the three-dimensional microvascular anatomy of the optic nerve and peripapillary choroid in the rat eye. METHODS: Gross vascular anatomy of the posterior eye segment of Wistar rats was studied in serial microsections with a light microscope. The optic nerve and peripapillary choroidal vessels were sequentially microdissected, using methylmethacrylate corrosion microvascular castings, and were examined with a scanning electron microscope to determine the three-dimensional relationships of the vessels. RESULTS: The posterior ciliary artery traveled along the inferior side of the optic nerve sheath, directly entered the optic nerve head, and divided into three branches: the central retinal artery and medial and lateral long posterior ciliary arteries, which provided several short branches to the choroid. The optic nerve head vasculature was consistently nourished by a recurrent arteriole from the central retinal artery and an arteriole from the choroidal artery at the peripapillary choroid. The central retinal vein flowed into a venous anastomosis along the optic disc border of the peripapillary choroid. Capillaries within the optic nerve drained into the central retinal vein, the marginal venous anastomosis of the peripapillary choroid, and the pial veins, all of which flowed into the posterior ciliary veins along the optic nerve sheath. CONCLUSIONS: The findings illustrate vascular anatomic differences in optic nerve and peripapillary choroidal microcirculation between rat and human. In rats, the peripapillary choroid plays a significant role in both blood supply and venous drainage of the optic nerve head. The central retinal artery also contributes to the optic nerve head circulation.

A novel RNA splicing mutation in Japanese patients with Wilson disease.

Deletion/insertion mutation of Wilson disease (WD) gene in 16 Japanese patients with Wilson disease was studied. A truncated size in a region of exon 4 to 6 was found by reverse transcription-polymerase chain reaction (RT-PCR) covering entire 21 exons except exon 1 for liver cDNA of one patient with a late onset neurologic type. Sequence analysis of the cDNA revealed that this truncation was occurred by skipping of exon 5, though any mutation in exon 5 of genomic DNA was failed to detect. T to G transversion in 5 bp upstream from a junction of intron 4 and exon 5 was found in genomic DNA of the patient. Further, results obtained by RT-PCR and the sequence analysis in intron 4 indicate that the mutation of the patient is homozygous. Since same mutation in one allele of another patient out of 15 patients was found, allele frequency of the splicing mutation in Japanese patients is 9.4%. These results suggest that the point mutation in intron 4 of WD gene causes the skipping of exon 5 and the splicing mutation affects the phenotype of Wilson disease.

[Statistical analysis of nuclear medicine in the Higashi Nippon Gakuen University Dental Hospital].

The number of radionuclide in vivo tests carried out at the Higashi Nippon Gakuen University Dental Hospital was statistically analyzed and discussed. The subjects were all patients admitted to the Division of Nuclear Medicine during the seven years between 1981 and 1989. The total number of the patients included those introduced through medical facilities in Toubetsu-cho and others. The results of this analysis were as follows: 1) The proportion of the first time patients admitted to the Higashi Nippon Gakuen University Dental Hospital was only 11 percent of all patients. 2) Cases of radionuclide in vivo tests at the Higashi Nippon Gakuen University Dental Hospital decreased in number, annually. However the proportion of tumor scintigraphy and bone scintigraphy increased gradually.

[Estimating organ doses of dental students in dental radiographic exercises].

To estimate doses absorbed in organs of dental students in the exercises of dental radiography, we measured the average organ doses of the dental radiographic examinations and sent out questionnaires about the radiographic exposure of the dental students. We selected five organs in the head and neck regions: lens of the eye, parotid gland, submaxillary gland, thyroid and pituitary fossa. The dose determinations were made for the full mouth intra-oral radiography with 10 films, and panoramic radiography and cephalometric radiography. Doses for all organs, except for the parotid gland, were widely distributed with one peak. The highest average dose per dental student was about 5000 microGy for the parotid gland, about 2900 microGy for the submaxillary gland, about 900 microGy for the lens of the eye, about 480 microGy for the thyroid and about 140 microGy for the pituitary fossa. Dose limits for dental students in such exercises have not been yet established. But the organ doses, especially for the parotid gland, appear very high. To reduce exposure in the exercises and to protect the dental students from excess exposure, we need to reconsider the study of dental radiographic techniques.

Evaluation of thyroid scintigraphy in small animals.

Thyroid scintigraphy in rats and mice with 99mTc and 123I was attempted to examine whether this modality might be used in small animals and to describe the possibilities of its application in examining experimentally produced thyroid diseases. A human gamma camera with a pinhole collimator improved for small animals was used for imaging. Thyroid images with 99mTc were obtained 20 minutes after injection and those with 123I were 24 hours after administration. All the scintigrams, although those obtained from mice with 99mTc displayed overlapped salivary gland images, demonstrated sufficiently clear images in size and shape, which showed that thyroid scintigraphy in small animals should be applicable in examining experimentally produced thyroid diseases.

[Experimental study on tooth fractures in young rats].

The influence of tooth fractures on growth of rat incisors was examined with soft X-ray radiography, xeroradiography and microradiography. Thirty young rats were used in this study and experimental fractures were produced on the mandibular left incisors. Radiological observations were carried out immediately, 7, 14, 21, 28 and 35 days after fracturing. The soft X-ray radiograms obtained immediately after fracturing showed clear radiolucent images. The radiopacity of the fractured regions gradually increased from the 7th day. Enamel hypoplasia was observed from the 7th day, and it moved towards the edge of the incisors with the growth of the fractured teeth. The xeroradiograms obtained at each term demonstrated as same findings as those obtained with soft X-ray radiography. The microradiograms obtained immediately after fracturing showed no noteworthy findings except for the clear radiolucent area of the fractured regions. Fine radiopaque lines along the edges of fractured regions were observed from the 7th day. Dentin-like radiopaque calcified masses were observed from the 14th day.