Expression Profiling of Calcium Induced Genes in Cultured Human Keratinocytes

Terminal differentiation of skin keratinocytes is a vertically directed multi-step process that is tightly controlled by the sequential expression of a variety of genes. To examine the gene expression profile in calcium-induced keratinocyte differentiation, we constructed a normalized cDNA library using mRNA isolated from these calcium-treated keratinocytes. After sequencing about 10,000 clones, we were able to obtain 4,104 independent genes. They consisted of 3,699 annotated genes and 405 expressed sequence tags (ESTs). Some were the genes involved in constituting epidermal structures and others were unknown genes that are probably associated with keratinocytes. In particular, we were able to identify genes located at the chromosome 1q21, the locus for the epidermal differentiation complex, and 19q13.1, another probable locus for epidermal differentiation-related gene clusters. One EST located at the chromosome 19q13.1 showed increased expression by calcium treatment, suggesting a novel candidate gene relevant to keratinocyte differentiation. These results demonstrate the complexity of the transcriptional profile of keratinocytes, providing important clues on which to base further investigations of the molecular events underlying keratinocyte differentiation.

Clinical Manifestations and Molecular Biologic Diagnosis of Herpetic Glaucoma

PURPOSE: We attempted to detect the herpes virus using a polymerase chain reaction (PCR) technique from the aqueous humor of patients who were suspected of having herpetic uveitis. METHODS: We obtained aqueous humor from four patients who suffered from elevated intraocular pressure and uveitis but did not respond to steroid and intraocular pressure depressants. We employed the PCR technique for to detect herpes-virus specific DNA. Patients were treated with oral and topical acyclovir. RESULTS: Uveitis of varying severity and progressive iris atrophy was observed in all patients. Varicella zoster virus (VZV) specific DNA was detected in two of four patients. Two patients showed a favorable course but the other two patients showed an unfavorable course due to advanced peripheral anterior synechia and corneal decompensation. CONCLUSIONS: Diagnosis of herpetic uveitis was difficult because it was not usually accompanied by dendritic corneal lesions. Herpetic uveitis should be considered whenever anterior uveitis and elevated intraocular pressure accompanies the characteristic iris atrophy. PCR was very useful in detecting the virus DNA in the aqueous humor to enable rapid diagnosis and adequate treatment.

Primary Cutaneous Anaplastic Large Cell Lymphoma of the Lower Lid

PURPOSE: Primary cutaneous anaplastic large cell lymphoma is rarely encountered in the lower eyelids. We report a patient with primary cutaneous anaplastic large cell lymphoma arising from the lower eyelid. METHODS: A 39-year-old man presented with a relatively fast growing mass on the center of his left lower eyelid for one month. The mass did not respond to local injection of triamcinolone at a local clinic. The lesion appeared as a solitary reddish nodule with ulceration, was non-tender, round, crusted, and measured 13 mm x 11 mm x 5 mm. Well- developed superficial vessels were found on the surface of the nodule. An incisional biopsy was performed. RESULTS: Histologic examination revealed that the bulk of the infiltrate was in the papillary and reticular dermis. Tumor cells had abundant, well-defined cytoplasm and pleomorphic nuclei with multiple nucleoli. The majority of the neoplastic cells showed immunoreactivity for CD 30 (Ki-1) along the cell membrane. A histopathological diagnosis of primary cutaneous anaplastic large cell lymphoma was made. CONCLUSIONS: Most cases of primary cutaneous anaplastic large cell lymphoma arise from the body and extremities. However, since primary cutaneous anaplastic large cell lymphoma may occur in the eyelid, it should be differentiated from nodular and relatively fast growing inflammatory tumors despite local steroid treatment.

Clinical Results of Deep Anterior Lamellar Keratoplasty

PURPOSE: To evaluate the surgical outcome of deep anterior lamellar keratoplasty (DALKP) in patients with corneal opacity with an intact corneal endothelium. METHODS: We transplanted partial thickness donor cornea, devoid of endothelium and Descemet's membrane, on to a recipient bed of which the deep stromal area had been dissected to expose the smooth surface of Descemet's membrane. We evaluated the visual acuity, refractive error, endothelial cell density, intraocular pressure, complication and graft rejection after a follow-up period of at least 6 months. RESULTS: Twelve eyes of 11 patients (M: F=7: 4) were enrolled in this study, with a mean age of 39 years (17~60). The mean follow-up period was 9.8 months. The underlying diseases were keratoconus in 6 eyes, corneal dystrophy in 4 and herpetic corneal lesion in 2. The mean postoperative visual improvement was 5 lines of Snellen chart. Microperforations occurred during operation in 4 eyes, and were treated with intracameral gas injection. One of them underwent penetrating keratoplasty due to persistent double anterior chamber. Transient elevation of intraocular pressure was noted in 7 eyes, but no case of graft rejection was observed during the follow-up period. CONCLUSIONS: Even though intraoperative microperforation may require an additional procedure such as gas injection which increases the possibility of intraocular pressure elevation, deep anterior lamellar keratoplasty is thought to be a good option for the primary surgery in patients with corneal opacity with an intact corneal endothelium due to its relatively fast recovery, reduced risk of rejection and preservation of endothelium without ongoing loss.

The Effects of Laser Photocoagulation on Reopened Macular Holes, as Assessed by Optical Coherence Tomography

PURPOSE: To evaluate the effects of laser photocoagulation on reopened macular holes. METHODS: This study involved 9 eyes from 9 patients who underwent laser photocoagulation coupled with fluid-gas exchange for reopened macular holes. The photocoagulation was performed at the center of the macular hole. Closure of the reopened hole was categorized by optical coherence tomography (OCT) according to the presence (type 1 closure) or absence (type 2 closure) of continuity in the foveal tissue. Best corrected visual acuity (BCVA), closure types, and complications were assessed. RESULTS: Upon final examination, all macular holes were found to have closed. Six eyes were classified as type 1 closure, and three were classified as type 2 closure. The mean BCVAs, before and after laser photocoagulation, were 0.11 and 0.31, respectively (P< .05). The eyes with type 1 closure were associated with shorter symptom durations and greater visual improvement than those with type 2 closure (P< .05). CONCLUSIONS: The combination of laser photocoagulation and fluid-gas exchange appears to be a safe and effective treatment for reopened macular holes. Early intervention should be encouraged to ensure complete hole closure and improved visual outcomes.

OPTN gene Mutation in Normal-Tension Glaucoma

PURPOSE: We have evaluated the mutations of the OPTN gene, which has been reported to be associated with the normal tension glaucoma (NTG). METHODS: The OPTN gene was analyzed in 53 patients with NTG and 40 normal subjects. Genomic DNA was extracted from the blood samples of each patients, exon 5 and exon 6 of the OPTN gene were amplified by PCR and DNA sequencing was performed. RESULTS: No mutation was found in normal subjects. But three kinds of point mutation (G412A, C459T in exon 5, G577C in exon 6) were found in 7 patients with NTG. CONCLUSIONS: We report the novel point mutations of OPTN gene in NTG patients. This shows the possibility of diagnosis of NTG by detecting the mutation of OPTN gene.