Kazuo Nakatsuka

Analysis of p53 Mutations and the Expression of p53 and p21WAF1/CIP1 Protein in 15 Cases of Sebaceous Carcinoma of the Eyelid

PURPOSE The purpose of this study was to detect mutation of the p53 gene, to assess its relationship with p53 or p21(WAF1/CIP1) expression, and to evaluate the correlation between p53 mutation or p21(WAF1/CIP1) expression and clinicopathologic findings in sebaceous carcinoma of the eyelid. METHODS Fifteen conventional paraffin-embedded samples of sebaceous carcinoma of the eyelid were analyzed. Using the single-strand conformation polymorphism technique, the authors sequenced coding exons 5-8 of the p53 gene. The expression of p53 and p21(WAF1/CIP1) protein was analyzed by immunohistochemistry. RESULTS In 10 of the 15 cases (66.7%), point mutations were detected in the p53 gene. CC to TT double-base changes (tandem mutations), which are known to be induced only by UV, were not detected in any of the mutations. Correlations between p53 mutation and expression were found to be statistically significant (P = 0.007). There was no significant correlation between p53 mutation and clinicopathologic findings or p21(WAF1/CIP1) expression. However, there was a significant inverse correlation between p21(WAF1/CIP1) expression and presence of lymph node metastasis (P = 0.007). CONCLUSIONS Among human cancers, sebaceous carcinoma of the eyelid may be one of those showing most frequent mutation of the p53 gene, which may not be caused by exposure to UV. p21(WAF1/CIP1) downregulation may be associated with lymph node metastasis.

Retinal circulatory disturbances following intracameral injection of bevacizumab for neovascular glaucoma.

acizumab injection affected the outcome in this case (TCVOS 1997). This is because ischaemic CRVO occurred 3 weeks after the injection, when bevacizumab has its peak effect and relatively good initial visual acuity and perfusion status. There were no other patient factors that might cause the ischaemic conversion of CRVO except diabetes. The causes and mechanisms of treatment failure associated with bevacizumab administration must be elucidated further in vitro and by clinical studies. Furthermore, the possibility that bevacizumab has a negative long-term effect on collateral vessel formation because of its anti-VEGF action requires investigation.

Changes in Refraction Caused by Induction of Acute Hyperglycemia in Healthy Volunteers

PURPOSE To determine whether the myopic changes and ocular hypotension after a glucose load are caused by hyperglycemia. METHODS Oral glucose tolerance tests were conducted on seven healthy young subjects with normal vision. The changes in the hematologic parameters and the refractive system were measured periodically for 150 minutes after the glucose load. RESULTS After the glucose load, there was an increase in plasma glucose level and the level of plasma osmosis, ocular hypotension, a myopic change in refractive power, shallowing of the anterior chamber, and a thickening of the lens. The degree of the myopic change exceeded the power of the residual accommodation. Normalization of the plasma glucose level led to a normalization of the intraocular pressure and a reversal of the myopic changes. CONCLUSIONS These findings suggest that the myopic changes that accompanied hyperglycemia were caused by a thickening of the lens resulting from a decrease in the tension of the zonule fibers of Zinn, and were secondary to ocular hypotension. Hyperopia appeared to be caused by the reversal of the myopia after normalization of plasma glucose levels.

Sebaceous Carcinoma Responds to Radiation Therapy

We describe a patient with an eyelid tumor that responded well to radiation therapy. The histopathological diagnosis was poorly differentiated sebaceous carcinoma. The patient received 52 Gy electron beam irradiation in a 5-week period; thereafter, the tumor diminished. The histopathological findings after radiation showed that most tumor cells underwent massive necrosis with hyalinized obstructive vessels. The tumor was under control at the 9-month follow-up examination.

The PI3K/Akt pathway mediates the expression of type I collagen induced by TGF-β2 in human retinal pigment epithelial cells.

PurposeTransforming growth factor (TGF)-β is a key mediator of proliferative vitreoretinopathy, but the cellular mechanisms by which TGF-β induces extracellular matrix protein (ECM) synthesis are not fully understood. This study examined whether the PI3K/Akt pathway is involved in TGF-β2-induced collagen expression in human retinal pigment epithelial cells.MethodsHuman retinal pigment epithelial cells ARPE-19 were cultured and stimulated with TGF-β2. The role of the PI3K/Akt pathway was evaluated using the biochemical inhibitor, wortmannin. The effect of wortmannin on the expression of type I collagen mRNA (COL1A1, COL1A2) induced by TGF-β2 was evaluated by real-time RT-PCR. The effect of wortmannin on the synthesis of type I collagen induced by TGF-β2 was assessed by an immunocytochemical analysis with anti-type I collagen antibody. Luciferase reporter assays were performed to examine the effect of wortmannin on the transcriptional activities of COL1A2. A luciferase assay using a mutation construct of the Smad binding site in COL1A2 promoter (Smad-mut/Luc) was also performed to examine the crosstalk between the Smad pathway and the PI3K/Akt pathway. The effects of wortmannin on the transcriptional activity of Smad3 were also examined using CAGA12-Luc. Moreover, the effect of wortmannin on TGF-β2-induced Smad7 mRNA expression was evaluated.ResultsThe biochemical blockade of PI3K/Akt activation inhibited TGF-β2-induced type I collagen mRNA expression and type I collagen synthesis. The blockade of PI3K/Akt pathway inhibited the increase in COL1A2 promoter activities when induced by TGF-β2 and reduced TGF-β2 induction of Smad-mut/Luc promoter activity and CAGA12-Luc activity. Moreover, wortmannin increased the TGF-β2-induced Smad7 mRNA expression levels.ConclusionsThe PI3K/Akt pathway plays a role in relaying the TGF-β2 signal to induce type I collagen synthesis in the retinal pigment epithelium through Smad-dependent and Smad-independent pathways.

Peribulbar fungal abscess and endophthalmitis following posterior subtenon injection of triamcinolone acetonide

for treatment of diabetic retinopathy. His best corrected visual acuity (VA) was 0.07 OD and 0.02 OS. He agreed to our recommendation that the macular oedema in the left eye be treated by a subtenon triamcinolone acetonide (TA) injection. The eye was anaesthetized with 4% topical lidocaine, and disinfected along with the lid margins and skin with 5% povidone iodine. A lid speculum was used, and the superonasal conjunctiva was incised to inject 20 mg of TA into the posterior subtenon space using a 27-gauge cannula with a dull tip. Topical ofloxacin ointment was instilled and levofloxacin q.i.d. was prescribed. Three months later, the patient reported diplopia and deep ocular pain OS. He had ptosis and restricted motility, and VA had dropped to 0.5 OS. The anterior chamber was quiet and the conjunctiva was not injected. Magnetic resonance imaging (MRI) showed a high-intensity lesion in the superonasal peribulbar area (Fig. 1A). Two months later, the diplopia worsened, and an epibulbar abscess was found superonasally (Fig. 1B). A repeat MRI revealed exophthalmos with two dome-like periocular cysts OS (Fig. 1C). The abscess was incised to drain a considerable amount of yellowishwhite purulent fluid. Gatifloxacin, pimaricin, levofloxacin (400 mg ⁄day) and itraconazole (100 mg ⁄day) were given topically and fluconazole (100 mg ⁄day) intravenously. Cultures yielded Scedosporium apiospermum, which was confirmed by DNA analysis. Therefore, itraconazole was replaced with voriconazole (300 mg ⁄day), to which the fungus was sensitive. The symptoms gradually decreased (Fig. 1D), but 4 weeks later the patient developed a central scotoma and a concentric visual field defect OS. His VA had decreased to 0.02. Repeat MRI showed a retrobulbar lesion in the superonasal region (Fig. 1E, F). Extensive vitritis with opacities, pale optic disc, periphlebitis, serous detachment of the macula, focal retinal haemorrhages and a whitish subretinal peripheral mass were noted (Fig. 1G–I). The former abscess, now replaced by fibrosis, was irrigated with balanced salt solution containing voriconazole, amphotericin B and ceftazidime. Six days later, pars plana vitrectomy and cataract extraction were performed. The mass gradually decreased, the retina was reattached, and VA improved to 0.5. No recurrence was observed in the following 6 months. Triamcinolone acetonide is a longacting steroid, and intravitreal or subtenon injection of TA has been used to treat macular diseases (Martidis et al. 2002). However, an intravitreal injection can cause a retinal detachment and endophthalmitis, and a subtenon TA injection can cause secondary ptosis and ocular infections (Engelman et al. 2004; Erol & Topbas 2006; Kusaka et al. 2007; Oh et al. 2007). A Medline search identified three patients with an orbital abscess or endophthalmitis following a subtenon TA injection (Engelman et al. 2004; Kusaka et al. 2007; Oh et al. 2007). One of these was caused by Pseudallescheria boydii and did not respond to intensive itraconazole therapy, leading to phthisis (Oh et al. 2007). The anamorph, S. apiospermum, is a filamentous fungus found in soil and polluted water. Its teleomorph is P. boydii, an emerging opportunistic fungus that usually affects immunocompromised hosts. Although the possibility of endogenous origin in the diabetes patient cannot be fully ruled out, the source of P. boydii in our case may have been direct contact with polluted Diagnosis ⁄ Therapy in Ophthalmology

Electroretinographic assessment of early changes in ocular siderosis

We examined a patient with an iron intraocular foreign body and recorded electroretinograms (ERGs) before and after the removal of the foreign body by vitrectomy. The amplitudes of the rod and cone ERGs and the oscillatory potentials (OPs) in the injured eye were reduced before the operation. In addition, the photopic on-responses (b wave) were more reduced than off-responses (d wave). One year after surgery, the amplitudes of the rod, cone and photopic on- and off-responses were markedly improved to within the low normal limit. However, the OP amplitudes remained unchanged with lower values. These findings suggest that iron retinotoxicity leads to a dysfunction of all layers but the changes may be reversible in the early period of the disease. The late period iron toxicity produces more severe damage to the inner retina than the outer retina.

Diagnosis of an isolated retinal astrocytic hamartoma aided by optical coherence tomography

girl was referred for evaluation of a retinal tumour in her left eye. Her visual acuity was 20 ⁄ 20 bilaterally. A solitary translucent retinal tumour measuring 1.5 · 3.0 disc diameters was seen inferior to the optic disc (Fig. 1A). The retinal arteries passing through the tumour appeared white. The sensitivity over the tumour was not altered in Humphrey visual field tests (Fig. 1B). Fluorescein fundus angiography (FA) showed retinal circulation to be intact with only weak staining of a section of the tumour (Fig. 1C,D). An ultrasound B-mode echography showed a mass inferior to the optic disc; no calcifications were observed (Fig. 1E). The fundus was scanned with optical coherence tomography (OCT) (3D OCT-1000; Topcon, Tokyo, Japan). The measurement beam was focused on the macular area with a system

Unusual Protrusion of Conjunctiva in Two Neonates with Harlequin Ichthyosis

Background: We present two patients who developed severe protrusion of the conjunctiva and chemosis secondary to Harlequin ichthyosis (HI). Case Reports: Case 1 was a male infant diagnosed with HI who had parchment-like appearance and conjunctival protrusion with severe chemosis. Case 2 was a female infant on whom HI had been suspected before birth through ultrasonography. She showed thickened skin over the entire body and conjunctival protrusion with severe chemosis. For both cases, a vitamin A derivative was applied and the hyperkeratotic layer was peeled off every day. Great care was taken to sterilize and moisten the ocular surface. The conjunctival protrusion gradually improved and other systemic conditions were successfully treated. HI is a rare condition, but affected infants are surviving longer than previously and hence guidelines for ocular management are now required. Conclusions: Gentle and patient debridement of the hyperkeratotic skin and moisturizing were important in treating the unusual conjunctival protrusion.

Optical coherence tomographic evaluation of the outer retinal architecture in Oguchi disease

BackgroundTo report the changes in the optical coherence tomography (OCT) images of the outer retinal layers after prolonged dark adaptation in a patient with Oguchi disease.CaseA 75-year-old woman showed the typical golden-yellow fundus reflex of Oguchi disease, and the coloration returned to normal after prolonged dark adaptation. Fourier domain OCT (FD-OCT) was performed on the patient’s left eye before and after prolonged dark adaptation.ObservationsBefore dark adaptation, the FD-OCT images at the fovea had three identifiable reflection bands, namely, the external limiting membrane (ELM), the border between the photoreceptor inner and outer segments (IS/OS line), and the retinal pigment epithelium (RPE)/Bruch band. The paramacular area had only the ELM and RPE/Bruch bands. After 4 h of dark adaptation, the IS/OS line was also detected in the paramacular area.ConclusionsThe absence of the IS/OS line in the extramacular regions in the partly dark adapted condition was most likely due to a defect in the rod photoreceptors of this area. The emergence of the IS/OS line after prolonged dark adaptation suggests that microarchitectural changes occur in the photoreceptors and that the changes may be correlated with the improvement of rod function.