Antiadenoviral effects of N-chlorotaurine in vitro confirmed by quantitative polymerase chain reaction methods

Eiichi Uchio1, Hirotoshi Inoue1, Kazuaki Kadonosono21Department of Ophthalmology, Fukuoka University School of Medicine, Fukuoka, Japan; 2Department of Ophthalmology, Yokohama City University Medical Center, Yokohama, JapanPurpose: Adenoviral keratoconjunctivitis is recognized as one of the major pathogens of ophthalmological nosocomial infection worldwide. N-Chlorotaurine (Cl–HN–CH2–CH2–SO3H, NCT) is the N-chloro derivative of the amino acid taurine, which is an oxidant produced by human granulocytes and monocytes during inflammatory reactions. Using conventional viral plaque assay, it was previously shown that NCT causes inactivation of several human adenovirus (HAdV) serotypes. In this study, we evaluated the antiadenoviral effect of NCT by quantitative polymerase chain reaction (PCR) methods.Methods: A549 cells were used for viral cell culture, and HAdV serotypes 3, 4, 8, 19, and 37 were used. After calculating 50% cytotoxic concentration (CC50) of NCT by MTS (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) method, HAdV was cultured with NCT for 7 days, and extracted adenoviral DNA was quantitatively measured by real-time PCR.Results: A statistically significant (P < 0.05) dose-dependent inhibition was indicated for all serotypes except HAdV type 4 (HAdV4), which was maximally inhibited by only ~50%. Among the serotypes, NCT was particularly effective against HAdV8, HAdV19a, and HAdV37. The 50% effective concentration (EC50) obtained by real-time PCR of NCT ranged between 49 and 256 µM. EC50 of NCT against HAdV3 was slightly higher than that against serotypes of species D. The selective index (CC50/EC50) ranged between 41 and 60 except for HAdV4 (11.5).Conclusions: These results show that NCT has an antiviral effect against most serotypes of human HAdV inducing keratoconjunctivitis, indicating its possible therapeutic use.Keywords: adenovirus, N-chlorotaurine, epidemic keratoconjunctivitis, antiviral agen

Neuroprotective response after photodynamic therapy: Role of vascular endothelial growth factor

Background: Anti-vascular endothelial growth factor (VEGF) drugs and/or photodynamic therapy (PDT) constitute current treatments targeting pathological vascular tissues in tumors and age-related macular degeneration. Concern that PDT might induce VEGF and exacerbate the disease has led us to current practice of using anti-VEGF drugs with PDT simultaneously. However, the underlying molecular mechanisms of these therapies are not well understood. Methods: We assessed VEGF levels after PDT of normal mouse retinal tissue, using a laser duration that did not cause obvious tissue damage. To determine the role of PDT-induced VEGF and its downstream signaling, we intravitreally injected a VEGF inhibitor, VEGFR1 Fc, or a PI3K/Akt inhibitor, LY294002, immediately after PDT. Then, histological and biochemical changes of the retinal tissue were analyzed by immunohistochemistry and immunoblot analyses, respectively. Results: At both the mRNA and protein levels, VEGF was upregulated immediately and transiently after PDT. VEGF suppression after PDT resulted in apoptotic destruction of the photoreceptor cell layer in only the irradiated area during PDT. Under these conditions, activation of the anti-apoptotic molecule Akt was suppressed in the irradiated area, and levels of the pro-apoptotic protein BAX were increased. Intravitreal injection of a PI3K/Akt inhibitor immediately after PDT increased BAX levels and photoreceptor cell apoptosis. Conclusion: Cytotoxic stress caused by PDT, at levels that do not cause overt tissue damage, induces VEGF and activates Akt to rescue the neural tissue, suppressing BAX. Thus, the immediate and transient induction of VEGF after PDT is neuroprotective

A consensus definition for lamellar macular hole

Background: A consensus on an optical coherence tomography (OCT) definition of lamellar macular hole (LMH) and similar conditions is needed. Methods: The panel reviewed relevant peer-reviewed literature to reach a consensus on LMH definition and to differentiate LMH from other similar conditions. Results: The panel reached a consensus on the definition of 3 clinical entities: LMH, epiretinal membrane (ERM) foveoschisis and macular pseudohole (MPH). LMH definition is based on 3 mandatory criteria and 3 optional anatomical features. The 3 mandatory criteria are the presence of irregular foveal contour, the presence of a foveal cavity with undermined edges, and the apparent loss of foveal tissue. Optional anatomical features include the presence of epiretinal proliferation, the presence of a foveal bump and the disruption of the ellipsoid line. ERM foveoschisis definition is based on 2 mandatory criteria: the presence of ERM and the presence of schisis at the level of Henle’s fiber layer. Three optional anatomical features can also be present: the presence of microcystoid spaces in the inner nuclear layer (INL), an increase of retinal thickness, and the presence of retinal wrinkling. MPH definition is based on 3 mandatory criteria and 2 optional anatomical features. Mandatory criteria include the presence of a foveal sparing ERM, the presence of a steepened foveal profile and an increased central retinal thickness. Optional anatomical features are the presence of microcystoid spaces in the INL and a normal retinal thickness. Conclusions: The use of the proposed definitions may provide uniform language for clinicians and future research

Optical coherence tomography-based consensus definition for lamellar macular hole.

BackgroundA consensus on an optical coherence tomography definition of lamellar macular hole (LMH) and similar conditions is needed.MethodsThe panel reviewed relevant peer-reviewed literature to reach an accord on LMH definition and to differentiate LMH from other similar conditions.ResultsThe panel reached a consensus on the definition of three clinical entities: LMH, epiretinal membrane (ERM) foveoschisis and macular pseudohole (MPH). LMH definition is based on three mandatory criteria and three optional anatomical features. The three mandatory criteria are the presence of irregular foveal contour, the presence of a foveal cavity with undermined edges and the apparent loss of foveal tissue. Optional anatomical features include the presence of epiretinal proliferation, the presence of a central foveal bump and the disruption of the ellipsoid zone. ERM foveoschisis definition is based on two mandatory criteria: the presence of ERM and the presence of schisis at the level of Henle's fibre layer. Three optional anatomical features can also be present: the presence of microcystoid spaces in the inner nuclear layer (INL), an increase of retinal thickness and the presence of retinal wrinkling. MPH definition is based on three mandatory criteria and two optional anatomical features. Mandatory criteria include the presence of a foveal sparing ERM, the presence of a steepened foveal profile and an increased central retinal thickness. Optional anatomical features are the presence of microcystoid spaces in the INL and a normal retinal thickness.ConclusionsThe use of the proposed definitions may provide uniform language for clinicians and future research

脈絡膜破裂のある外傷性黄斑円孔への硝子体手術の1例

背景:脈絡膜破裂を伴った外傷性黄斑円孔に対し硝子体手術を行った一例を経験したので報告する.症例と所見:26歳女性,うちあげ花火が直接右眼にあたり鈍的外傷となった.初診時の矯正視力は0.01であり,白内障,前房出血,黄斑円孔,脈絡膜破裂に伴う黄斑下出血と網膜浮腫がみられた.3週間後黄斑円孔は拡大した.6週後の蛍光眼底造影にて黄斑部に及ぶ網膜循環障害を認めた.受傷6週間後にインドシアニングリーンを用いた内境界膜剥離術を行った.黄斑下手術は行わなかった.結果:術後黄斑円孔は閉鎖し,視力は0.5に改善した.重篤な術後合併症は見られなかった.結論:脈絡膜破裂のある外傷性黄斑円孔に対しても硝子体手術は有効であると考えられた.Background: We report a case of traumatic macular hole with choroidal rupture treated with vitrectomy. Case & findings: The patient was a 26-years-old female, who was hit by fireworks in the right eye. At the initial visit, objective symptoms were cataract, hyphema, macular hole, retinal edema, and subretinal hemorrhage following choroidal rupture. Best corrected visual acuity was 0.01 in the right eye. A macular hole developed within 3 weeks, and retinal circulatory disorder was detected in fluorescein angiography. 6 weeks later, pars plana vitrectomy with internal limited membrane peeling using indocyanine green was performed. Sub-retinal procedure in the macular was not included in the surgery. After the surgery, the macular hole was closed, and the best corrected visual acuity in the right eye improved to 0.5. No severe complications were seen postoperatively. Conclusion: Vitreous surgery was effective for traumatic macular hole with choroidal rupture

Intra-retinal Arterial Cannulation using a Microneedle for Central Retinal Artery Occlusion

Abstract Central retinal artery occlusion (CRAO) is a severe disease, often causing blindness. We evaluated the efficacy and safety of a surgical procedure for the treatment of acute CRAO in which retinal arterial cannulation with tissue plasminogen activator (tPA) is performed. The surgical procedure consisted of vitrectomy followed by cannulation of the central retinal artery and injection of tPA (200 μg) using a 47-gauge microneedle. Thirteen CRAO patients were treated within 48 hours of the onset of symptoms. The central retinal artery of all 13 eyes was successfully cannulated. The mean interval between the onset of symptoms and surgery was 38.7 hours. The results for all 13 eyes treated showed a statistically significant improvement in mean visual acuity between before and one month after treatment (−1.60 vs. −0.82 logarithmic values for minimum angle resolution (LogMAR), p = 0.0021). Fluorescein angiography showed complete reperfusion and incomplete reperfusion in 10 eyes and 3 eyes, respectively. Recently developed surgical instruments have made retinal-arterial cannulation feasible. Intra-retinal-arterial cannulation has potential as a method of improving visual function and microcirculation in eyes affected by CRAO