Effect of Orthokeratology Lens on Contrast Sensitivity Function and High-Order Aberrations in Children and Adults.

OBJECTIVES: Overnight orthokeratology (OOK) lenses are used to temporarily decrease myopic refractive error and improve uncorrected vision. Overnight orthokeratology lenses significantly increase ocular and corneal high-order aberrations (HOAs) and compromise contrast sensitivity function (CSF) to a degree correlated with myopic correction achieved. In Taiwan, OOK lenses are mainly used in children for myopia control. However, information regarding its effects in this population remains limited. This study discusses the change in HOAs and CSF after 28 nights of OOK lens use in children compared with that in adults. METHODS: In total, 46 children (9-18 years) and 26 adults (>18 years) who visited Ophthalmology Department of Mackay Memorial Hospital from October to December 2013 were enrolled. Contrast sensitivity and ocular/corneal total high-order, coma, and spherical aberrations (SA) were tested before OOK treatment. After 28 days of overnight use, CSF and topography were reexamined, and data were collected and analyzed using t test and Pearson correlation coefficients. RESULTS: In total, 23 eyes of 23 children and 14 eyes of 14 adults were evaluated. The treatment resulted in a significant increase in ocular total HOA, coma, and SA in both groups. However, CSF declined more in adults than children. CONCLUSION: Our study revealed that OOK lenses decrease CSF to a greater extent in adults than that in children despite no significant differences in the change of ocular HOAs between both subject groups. We proposed children may have better neural adaptation to compensate for optical aberrations induced by OOK lens use.

PEDF promotes self-renewal of limbal stem cell and accelerates corneal epithelial wound healing.

Limbal epithelial stem cell (LSC) transplantation is a prevalent therapeutic method for patients with LSC deficiency. The maintenance of stem cell characteristics in the process of culture expansion is critical for the success of ocular surface reconstruction. Pigment epithelial-derived factor (PEDF) increased the numbers of holoclone in LSC monolayer culture and preserved the stemness of LSC in suspension culture by evidence of ΔNp63α, Bmi-1, and ABCG2 expression. BrdU pulse-labeling assay also demonstrated that PEDF stimulated LSCs proliferation. In air-lift culture of limbal equivalent, PEDF was capable of increasing the numbers of ΔNp63α-positive cells. The mitogenic effect of PEDF was found to be mediated by the phosphorylations of p38 MAPK and STAT3 in LSCs. Synthetic 44-mer PEDF (residues 78-121) was as effective as the full length PEDF in LSC expansion in suspension culture and limbal equivalent formation, as well as the activation of p38 MAPK and STAT3. In mice subjecting to mechanical removal of cornea epithelium, 44-mer PEDF facilitated corneal wound healing. Microscopically, 44-mer PEDF advanced the early proliferative response in limbus, increased the proliferation of ΔNp63α-positive cells both in limbus and in epithelial healing front, and assisted the repopulation of limbus in the late phase of wound healing. In conclusion, the capability of expanding LSC in cell culture and in animal indicates the potential of PEDF and its fragment (e.g., 44-mer PEDF) in ameliorating limbal stem cell deficiency; and their uses as therapeutics for treating corneal wound.

Inhibition of microRNA-328 Increases Ocular Mucin Expression and Conjunctival Goblet Cells.

We previously reported anti-miR-328 therapy for dry eye disease (DED). Since decreased mucin secretion is a risk factor for DED, we aimed to explore whether anti-miR-328 affects mucin expression and goblet cells. MiR-328 was increased in goblet cells when they were under desiccating stress or treated with benzalkonium chloride (BAC), both of which are risk factors for DED. Based on bioinformatics tool results, miR-328 was predicted to directly target the transcription factor CREB1 that has been known to promote the expression of mucin5AC. The inhibitory effect of miR-328 on CREB1 was confirmed by the transfection assay. A miR-328 binding site on the CREB1 gene was confirmed by the luciferase assay. Furthermore, anti-miR-328 increased CREB1 and mucin5AC in cultured goblet cells according to qPCR, Western blot, and IF staining experiments. Anti-miR-328 increased mucin5AC secretion from the cultured goblet cells based on an ELISA assay for the cultured medium. Finally, impression cytology data revealed anti-miR-328 increased conjunctival goblet cells in the DED rabbits induced by BAC. In conclusion, anti-miR-328 increases CREB1 expression leading to an increase in mucin5AC production and secretion. Furthermore, anti-miR-328 also increases conjunctival goblet cells. These results warrant the further development of anti-miR-328 therapy for DED.

Pigment epithelium-derived factor (PEDF) promotes tumor cell death by inducing macrophage membrane tumor necrosis factor-related apoptosis-inducing ligand (TRAIL).

Pigment epithelium-derived factor (PEDF) is an intrinsic anti-angiogenic factor and a potential anti-tumor agent. The tumoricidal mechanism of PEDF, however, has not been fully elucidated. Here we report that PEDF induces the apoptosis of TC-1 and SK-Hep-1 tumor cells when they are cocultured with bone marrow-derived macrophages (BMDMs). This macrophage-mediated tumor killing is prevented by blockage of TNF-related apoptosis-inducing ligand (TRAIL) following treatment with the soluble TRAIL receptor. PEDF also increases the amount of membrane-bound TRAIL on cultured mouse BMDMs and on macrophages surrounding subcutaneous tumors. PEDF-induced tumor killing and TRAIL induction are abrogated by peroxisome proliferator-activated receptor γ (PPARγ) antagonists or small interfering RNAs targeting PPARγ. PEDF also induces PPARγ in BMDMs. Furthermore, the activity of the TRAIL promoter in human macrophages is increased by PEDF stimulation. Chromatin immunoprecipitation and DNA pull-down assays confirmed that endogenous PPARγ binds to a functional PPAR-response element (PPRE) in the TRAIL promoter, and mutation of this PPRE abolishes the binding of the PPARγ-RXRα heterodimer. Also, PPARγ-dependent transactivation and PPARγ-RXRα binding to this PPRE are prevented by PPARγ antagonists. Our results provide a novel mechanism for the tumoricidal activity of PEDF, which involves tumor cell killing via PPARγ-mediated TRAIL induction in macrophages.

Pigment epithelium-derived factor is an intrinsic antifibrosis factor targeting hepatic stellate cells.

The liver is the major site of pigment epithelium-derived factor (PEDF) synthesis. Recent evidence suggests a protective role of PEDF in liver cirrhosis. In the present study, immunohistochemical analyses revealed lower PEDF levels in liver tissues of patients with cirrhosis and in animals with chemically induced liver fibrosis. Delivery of the PEDF gene into liver cells produced local PEDF synthesis and ameliorated liver fibrosis in animals treated with either carbon tetrachloride or thioacetamide. In addition, suppression of peroxisome proliferator-activated receptor gamma expression, as well as nuclear translocation of nuclear factor-kappa B was found in hepatic stellate cells (HSCs) from fibrotic livers, and both changes were reversed by PEDF gene delivery. In culture-activated HSCs, PEDF, through the induction of peroxisome proliferator-activated receptor gamma, reduced the activity of nuclear factor-kappa B and prevented the nuclear localization of JunD. In conclusion, our observations that PEDF levels are reduced during liver cirrhosis and that PEDF gene delivery ameliorates cirrhosis suggest that PEDF is an intrinsic protector against liver cirrhosis. Direct inactivation of HSCs and the induction of apoptosis of activated HSCs may be two of the mechanisms by which PEDF suppresses liver cirrhosis.

Effects of tremella-alginate-liposome encapsulation on oral delivery of inactivated H5N3 vaccine.

In this study, we evaluated a system for oral vaccine delivery, consisting of liposomes coated first with a layer of tremella and then with an outer layer of acid-induced alginate. In vitro release studies showed that the triple layer of alginate-tremella-liposomes was more resistant to an acidic pH and modulated the release profiles at an alkaline pH. Transepithelial electrical resistance (TEER) studies revealed that liposomes or tremella-coated liposomes were able to open tight junctions of the Caco-2 cell monolayer. In mice, although serum immunoglobulin G (IgG) was not expected to increase and haemagglutination inhibition showed that antibody levels were still too low to provide sufficient protection, alginate-tremella-liposomes encapsulated virus-induced intestinal secretory immunoglobulin A (s-IgA) production to provide protection against virus infection. In conclusion, an oral virus vaccine entrapped in alginate-tremella-liposomes improved the mucosal antiviral s-IgA response. This system may have potential use as a carrier for oral vaccine delivery.

The Use of Autologous Serum to Reverse Severe Contact Lens-induced Limbal Stem Cell Deficiency.

PURPOSE: To describe the efficacy of autologous serum (AS) eye drops to reverse severe contact lens (CL)-induced limbal stem cell (LSC) deficiency (LSCD). METHODS: This is a prospective, uncontrolled, interventional case series that enrolled 20 eyes of 14 consecutive patients diagnosed with severe CL-induced LSCD at presentation, based on clinical examination, at a tertiary referral center for the period December 2016 to December 2018. All eyes underwent AS treatment for at least 2 weeks with a follow-up for at least 2 months. Demographic data and treatment outcomes were collected and analyzed. RESULTS: The mean patient age at presentation was 30.5 years (range, 19-49 years). The mean duration of soft contact lens wear was 15.6 years (SD, 7.58 years; range, 5-31 years). All study eyes had pain and blurred vision at presentation. All eyes had recurrent or persistent corneal epithelial defect, stromal scarring and opacity, and superficial vascularization and peripheral pannus at presentation. Aggressive treatment with AS succeeded in all eyes. Signs and symptoms of LSCD stabilized in all eyes within 2 weeks and resolved in 6 eyes (30.0%) in 2 weeks, 9 eyes (45.0%) in 4 weeks, and 5 eyes (25.0%) in 8 weeks. The mean follow-up time was 9.45 ± 1.79 weeks (range, 8-24 weeks). CONCLUSIONS: Early identification and aggressive treatment of the ocular surface disease with AS can medically reverse severe CL-induced LSCD and prevent the need for surgical intervention.

Cytosolic phospholipase A2-{alpha} is an early apoptotic activator in PEDF-induced endothelial cell apoptosis.

Pigment epithelium-derived factor (PEDF) is an intrinsic antiangiogenic factor and a potential therapeutic agent. Previously, we discovered the mechanism of PEDF-induced apoptosis of human umbilical vein endothelial cells (HUVECs) as sequential induction/activation of p38 mitogen-activated protein kinase (MAPK), peroxisome proliferator-activated receptor gamma (PPAR-gamma), and p53. In the present study, we investigated the signaling role of cytosolic calcium-dependent phospholipase A(2)-alpha (cPLA(2)-alpha) to bridge p38 MAPK and PPAR-gamma activation. PEDF induced cPLA(2)-alpha activation in HUVECs and in endothelial cells in chemical burn-induced vessels on mouse cornea. The cPLA(2)-alpha activation is evident from the phosphorylation and nuclear translocation of cPLA(2)-alpha as well as arachidonic acid release and the cleavage of PED6, a synthetic PLA(2) substrate. Such activation can be abolished by p38 MAPK inhibitor. The PEDF-induced PPAR-gamma activation, p53 expression, caspase-3 activity, and apoptosis can be abolished by both cPLA(2) inhibitor and small interfering RNA targeting cPLA(2)-alpha. Our observation not only establishes the signaling role of cPLA(2)-alpha but also for the first time demonstrates the sequential activation of p38 MAPK, cPLA(2)-alpha, PPAR-gamma, and p53 as the mechanism of PEDF-induced endothelial cell apoptosis.

Pigment epithelium-derived factor induces THP-1 macrophage apoptosis and necrosis by the induction of the peroxisome proliferator-activated receptor gamma.

Pigment epithelial-derived factor (PEDF) is a potent anti-angiogenic factor, partially through the induction of endothelial cell apoptosis. Here we report that PEDF can also induce the apoptosis of human THP-1 monocytic leukemia cell line-derived macrophage cells (THP-1 macrophages) and peroxisome proliferator-activated receptor gamma (PPARgamma), a pleiotropic transcriptional factor is involved in the signaling. TUNEL and propidium iodide permeability assays demonstrated that PEDF dose- and time-dependently induces both apoptosis and necrosis of THP-1 macrophages while inducing the cleavages of procaspase-9, -3, the release of cytochrome c and the overexpression of p53. All these PEDF effects can be attenuated by either PPARgamma inhibitor GW9662 or PPARgamma small interfering RNA. The effects of PEDF can be reproduced by transient expression of PPARgamma by a PPARgamma-expression plasmid transfection. PEDF increased the expression and transcriptional activity of PPARgamma in THP-1 macrophages. In addition, PEDF also induced apoptosis in primary human monocyte-derived macrophages (MDMs) while inducing the expression of PPARgamma. Our observations indicate that PEDF induces macrophage apoptosis and necrosis through the signaling of PPARgamma. This suggests a novel mechanism through which PEDF can modulate inflammation.

Troglitazone suppresses transforming growth factor beta-mediated fibrogenesis in retinal pigment epithelial cells.

PURPOSE: Transforming growth factor (TGF)-beta2 induction of epithelial-mesenchymal transition of retinal pigment epithelium (RPE) cells has been implicated to be an important event during the development of proliferative vitreoretinopathy. The present study was conducted to examine whether troglitazone (TGZ) can inhibit TGFbeta2-mediated fibrosis of RPE cells. The mechanism of the TGZ effect was also investigated by studying major TGFbeta2-induced signaling including activation of Smad and p38 mitogen activated protein kinase (MAPK). METHODS: Human RPE cells (ARPE-19) were exposed to various concentrations of TGZ in the presence of TGFbeta2. The inhibitory effects of TGZ on collagen type I (COLI) and fibronectin (FN) expression induced by TGFbeta2 was evaluated by reverse transcriptase-polymerase chain reaction. COLI synthesis was evaluated by the concentration of the C-terminal propeptide of COLI in the medium. The protein levels of FN and the phosphorylation of p38 MAPK and Smad2 and Smad3 were assessed by immunoblotting. TGZ inhibition of TGFbeta2-promoted ARPE-19 cell migration was evaluated by an in vitro wound-healing assay. The influence of TGZ on cell viability was evaluated by the colorimetric conversion of 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide. RESULTS: TGZ dose-dependently inhibited TGFbeta2-induced COLI and FN overexpression at the levels of mRNA and protein manufacture. A dose-dependent TGZ inhibition was also apparent in TGFbeta2-induced cell migration; cell viability was unaffected. TGFbeta2 induced sequential phosphorylation of Smad2 and Smad3 and p38 MAPK. TGZ inhibited TGFbeta2-induced early Smad2 and Smad3 and late Smad3 phosphorylation but had no influence on TGFbeta2-induced p38 MAPK activation. CONCLUSIONS: TGZ pretreatment can significantly prevent TGFbeta2-induced epithelial- mesenchymal transition of RPE cells, and retards cell migration. This may be achieved through the prevention of TGFbeta2-induced Smad2 and Smad3 phosphorylation and subsequent nuclear accumulation. On the other hand, TGZ does not alter the levels of TGFbeta2-induced p38 MAPK phosphorylation, the effect of TGZ is unlikely to be mediated by p38 MAPK signaling.

Cryopreservation of human limbal stem cells ex vivo expanded on amniotic membrane.

PURPOSE: After cornea transplantation, the donor's limbal zone is currently discarded as medical waste. However, the limbal zone is rich in limbal stem cells and can be used in therapeutic applications of limbus loss. This study aimed to increase the availability of limbal stem cells and develop the optimal conditions of cryopreservation for ex vivo expanded limbal stem cells. METHODS: Pieces of the limbus were cultured on amniotic membrane (AM) to outgrow limbal stem cells as cell sheets for 3 weeks. Different formulas of cryoprotectants were tested to preserve the expanded cell sheets in liquid nitrogen. Before and after cryopreservation, expanded cell sheets were assessed for cellular characteristics by viability, histologic examination, and expression of ABCG2, vimentin, and keratin 3. RESULTS: Expanded cell sheets usually exhibited 3-6 stratified layers after 3-week culture on AM and expressed specific markers of ABCG2 and vimentin for limbal stem cells. The effects of cryopreservation with different cryoprotectants were analyzed by histopathology, stem cell markers, and cell viability. The results showed that the optimal formula of cryoprotectants for expanded limbal cell sheets was 60% Dulbecco modified Eagle medium, 30% fetal bovine serum, and 10% dimethyl sulfoxide. After 8-week cryopreservation in liquid nitrogen, the characteristics of limbal stem cells were maintained, and the average viability of thawed cells was 53.8% +/- 5.8%. CONCLUSIONS: These results showed that limbal stem cells expanded on AM could be cryopreserved and provide a promising source without delay, if banking, for patients with limbal stem cell deficiency in the future.

Microkeratome blades and corneal flap thickness in LASIK.

BACKGROUND AND OBJECTIVE: To determine the corneal flap thickness produced by using two different types of blades on the same microkeratome for LASIK, comparing flap thickness in the first and second eyes. PATIENTS AND METHODS: The corneal flap thickness was measured in two sets of 100 consecutive patients undergoing bilateral LASIK procedures using the Moria M2 microkeratome with a 110 head (Moria, France) and either Moria or CLB blades (Med-Logics Inc., Laguna Hills, CA). Corneal flap thickness was determined by intraoperative pachymetry using an ultrasonic pachymeter. RESULTS: The mean corneal flap thickness was 138.95 +/- 21.6 microm (range: 75-205 microm) with Moria blades and 115.07 +/- 16.0 microm (range: 70-153 microm) with CLB blades, which is a significant difference (P < .001). The difference in flap thickness between the first and second eyes was not significant with the Moria blades (141.46 +/- 21.46 microm vs 136.45 +/- 21.55 microm, P = .965), but it was when the CLB blades were used (123.21 +/- 12.07 microm vs 106.93 +/- 15.51 microm; P= .013). CONCLUSION: In LASIK surgery using a Moria M2 microkeratome, blades from different manufacturers may produce significantly different corneal flap thicknesses, as well as differences between the first and second eyes.

Errors of residual stromal thickness estimation in LASIK.

BACKGROUND AND OBJECTIVE: To investigate inaccuracy and variability in residual stromal thickness estimation in LASIK by pachymetry and measurements of corneal thickness, flap thickness, and ablation depth. PATIENTS AND METHODS: In 73 eyes of 37 patients, preoperative and postoperative corneal thicknesses were obtained with slit-scanning elevation topography and the ultrasound pachymeter. LASIK was performed and corneal flaps were created with a microkeratome. Flap thickness and ablation depth (expected and achieved) were calculated. Residual stromal thickness estimation error was analyzed. RESULTS: The mean preoperative corneal thicknesses were 559.58 +/- 23.47 and 554.92 +/- 29.95 microm for the ultrasound pachymeter and slit-scanning elevation topography, respectively. Measurement differences ranged from -36 to 30 microm. With the pachymeter, calculated mean flap thickness was 139.58 +/- 17.59 microm. With this device, predicted ablation depth differed from achieved depth by 20% or more in approximately one-third (30.14%) of treated patients; ablation differences ranged from 10.0% to 19.99% in 37% of patients and 1.00% to 9.99% in 31.5% of patients. CONCLUSION: Imprecision of microkeratome cuts, preoperative corneal pachymetry, and laser ablation depth have a significant impact on the inaccuracy of residual stromal thickness prediction. Especially in patients with borderline corneal thickness, intraoperative pachymetry measurements and a residual stromal thickness higher than the safety margin of 250 microm are recommended to minimize iatrogenic ectasia.

Subconjunctival injection of recombinant AAV-angiostatin ameliorates alkali burn induced corneal angiogenesis.

PURPOSE: To evaluate the effect of subconjunctival injection of recombinant adeno-associated virus (rAAV)-angiostatin in alkali burn-induced corneal angiogenesis. METHODS: Adeno-associated viral vector-mediated gene delivery into extraocular tissue was determined by fluorescent microscopy three weeks after subconjunctival injection of viral vector expressing green fluorescent protein (rAAV-GFP). Subconjunctival injection of recombinant adeno-associated viral vector expressing human angiostatin (rAAV-angiostatin) and blank rAAV viral vector (control) was performed in the left eye of male Sprague-Dawley rats (n=6). Alkaline induction of corneal neovascularization (NV) was performed three weeks later. Corneal NV regression was analyzed 7-14 days after alkali burn with slit lamp and digital pictures. Transgenic expression of angiostatin in the cornea, conjunctiva, retina, and muscle insertions was confirmed by reverse-transcriptase polymerase chain reaction (RT-PCR). RESULTS: Transgenic GFP gene expression was detected mainly in the muscle fibers at the extraocular muscle (EOM) insertions after subconjunctival injection. The area of corneal neovascularization was significantly lower in eyes injected with rAAV-angiostatin (p<0.01) than in eyes injected with the control virus. RT-PCR demonstrated that the angiostatin gene expression was highly detectable in muscle fibers and not detectable in the conjunctiva, cornea, and retina. CONCLUSIONS: Subconjunctival injection of rAAV-angiostatin reduced alkali burn-induced corneal angiogenesis. We proved the concept that ocular gene therapy by subconjunctival injection of adenovirus-associated gene transfer of angiogenesis inhibitors can be a simple and safe treatment modality that can achieve therapeutic levels and long lasting effects in the treatment of corneal NV induced by ocular surface disorders.

Pigment epithelium-derived factor inhibits oxidative stress-induced cell death by activation of extracellular signal-regulated kinases in cultured retinal pigment epithelial cells.

Oxidative stress-induced retinal pigment epithelial (RPE) cell death is involved in the pathogenesis of age-related macular degeneration (AMD). Pigment epithelium-derived factor (PEDF) is an anti-angiogenic/neurotropic dual functional factor, and recently it was also shown to mediate anti-oxidative action. In the present study, the influence of PEDF in hydrogen peroxide (H(2)O(2))-induced RPE cell death was investigated using nontransformed human RPE cell line (ARPE-19). The recombinant PEDF was purified from E. coli. The MTT cell viability assay showed that PEDF rescued ARPE-19 from H(2)O(2)-induced cell death in a dose- and time-dependent manner. Western blot analysis revealed that PEDF stimulated the extracellular signal-regulated kinases (ERK1/2) phosphorylation. The PEDF cytoprotective effect was significantly attenuated by the ERK1/2 inhibitor PD98059. In this study, we demonstrate that PEDF induces ERK1/2 phosphorylation and we further suggest that the ERK signal cascade contributes to RPE cell's cytoprotection against oxidative stress.

Pro-oxidant and cytotoxic activities of atractylenolide I in human promyeloleukemic HL-60 cells.

The dried rhizome of Bai Zhu (Atractylodes ovata) is widely used as a Chinese herbal medicine. Two sesquiterpenolides of similar structures (atractylenolide I, AT-I; atractylenolide III, AT-III) were isolated from dried rhizome of Atractylodes ovata. Incubation of AT-I with recombinant human Cu,Zn-superoxide dismutase (rhCu,Zn-SOD) resulted in rhCu,Zn-SOD fragmentations and Zn releases. However, these were not observed in the AT-III reaction. The AT-1 showed dose-dependent cytotoxic activities (7.5, 15, and 30 microg/ml) on the human promyeloleukemic HL-60 cells while AT-III did not, and the IC50 of the former being 10.6 microg/ml (corresponding to 46 microM) on 12 h-treated cells. The results of DNA ladder and DNA contents in sub-G1 type revealed that AT-I induced apoptosis in human promyeloleukemic HL-60 cells. The cytotoxic and pharmacological mechanisms of AT-I against human promyeloleukemic HL-60 cells was investigated. The AT-I appeared to exhibit both pro-oxidant and antioxidant properties after an ESR spectrometer was used to detect hydroxyl radical productions in vitro and flow cytometry to detect intracellular ROS productions in AT-I treated cells. The AT-1 also showed dose-dependent Cu,Zn-SOD inhibitory activity in HL-60 cells treated for 12 h, confirmed by activity and immune stainings. However, catalase, Mn-SOD, and glutathione peroxidase did not apparently change activities under the same treatments. The addition of commercial rhCu,Zn-SOD (25-100 U/mL) to the AT-I-treated HL-60 cells (15 microg/ml) resulted in significant differences (p<0.01) and could reduce the AT-I cytotoxicity from 78% to 28% on HL-60 cells. It was proposed that the AT-I might work via Cu,Zn-SOD inhibition in HL-60 cells to induce apoptosis and bring about cytotoxicity.

Effectiveness and safety of overnight orthokeratology with Boston XO2 high-permeability lens material: A 24 week follow-up study.

OBJECTIVE: To examine the effectiveness of overnight orthokeratology lenses made with Boston XO2, highly gas-permeable lens material for the temporary correction of myopia. METHODS: Myopic individuals from 9 to 62 years of age were eligible. Participants ≤ 12 years of age were required to have myopia ≤-4.00 D and astigmatism ≤ 1.50 D, and for those 13-62 years of age myopia ≤-5.00 D and astigmatism ≤ 3.00 D. All participants were required to have normal healthy eyes and not be receiving any ocular medications or systemic medications likely to affect the results of visual acuity. Participants wore the lenses for a minimum of 7h during sleep, and were evaluated on day 1 and weeks 1, 2, 4, 12, and 24. Success was defined as LogMAR ≤ 0.1. RESULTS: A total of 126 participants (63.5% females) with a mean age of 20.4 ± 11.5 years were recruited. Baseline LogMAR, and vertical and horizontal corneal curvature were 0.8, 7.7 mm, and 7.9 mm, respectively, in both eyes. A consistent decrease in LogMAR was noted from day 1 to week 12. The success rate increased with length of time (from 33.9% to 100% for the right eye and from 35.5% to 100% for the left eye from day 1 to week 24). No severe complications were noted. CONCLUSION: Overnight orthokeratology with lenses made of Boston XO2 material are effective and safe for the temporary reduction of myopia.

Pigment Epithelial-Derived Factor Peptide Regenerated Limbus Serves as Regeneration Source for Limbal Regeneration in Rabbit Limbal Deficiency.

PURPOSE: To demonstrate that a 44-amino acid peptide from pigment epithelial-derived factor (PEDF) induces the regeneration of limbal excision wound, and the regenerated limbus can act as the regeneration source for new limbal excisional injuries in rabbit model of limbal deficiency. METHODS: Half circumference partial limbal excision was followed by PEDF peptide treatment to achieve limbal wound regeneration. Three months later, a second stage half circumference partial limbal excision removed the remaining native limbal tissue followed by PEDF peptide treatment. The structure and function of the regenerated limbus were analyzed at 3 and 6 months. Conjunctivalization was analyzed by impression cytology. Immunohistochemical analysis was performed with antibodies to corneal epithelium-associated keratin 3 (K3), conjunctival epithelium-associated keratin 13 (K13), ΔNp63α, ABCG2, and BrdU. Extensive limbal excision was performed to examine the regeneration potential of the PEDF peptide. RESULTS: Total limbal stem cell deficiency occurred with severe inflammation and conjunctivalization of the limbal wound and adjacent cornea in vehicle control eyes. In PEDF peptide treated eyes, the regenerated limbus prevented fibrovascular invasion and goblet cell migration into the corneal surface. Immunohistochemical staining of the regenerated limbus showed a wide distribution of cells expressing ΔNp63α and ABCG2 as in the native limbus. BrdU labeling assay revealed the presence of slow-cycling cells in the basal layer of the regenerated limbus. The PEDF peptide can heal extensive limbal excisional wounds and sustain ocular surface integrity. CONCLUSIONS: The addition of PEDF peptide has the potential to repair limbal excisional wounds with the recovery of normal limbus-like anatomy and function. The PEDF peptide is a potential remedy for extensive limbal injury.

Immobilized zinc affinity chromatography of pectin hydroxamic acids for purification of trypsin inhibitors from soybean and sweet potato.

Commercial pectin (with a 94% degree of esterification, DE94) suspended in methanol was reacted with methanolic alkaline hydroxylamine at room temperature for 20 h to prepare pectin hydroxamic acids (PHAs). The prepared PHA was coupled to the epoxy-activated Sepharose 6B gel to get immobilized PHA resins. The immobilized PHA resin was then balanced in column with 2 mM ZnCl2 in 50 mM Tris-HCl buffer (pH 7.9) to test the immobilized Zn-PHA gel as solid phase for immobilized metal affinity chromatography for the purification of trypsin inhibitors (TIs) from soybean and sweet potato. Using TI activity staining, it was found that purified TIs from the commercial soybean and sweet potato after trypsin affinity column purification could be adsorbed onto an immobilized Zn-PHA affinity column and eluted by 100 mM EDTA in 10 mM Tris-HCl buffer (pH 7.9). The immobilized Zn-PHA affinity column was used for TI purifications from crude extracts of sweet potato. The recovery of TI activity for one step was 90%, with 19.74-fold purification increase.

Distribution of corneal and ocular spherical aberrations in eyes with cataract in the Taiwanese population.

PURPOSE: To investigate the distribution of corneal and ocular spherical aberrations (SAs) in eyes with cataract in the Taiwanese population. METHODS: Corneal and ocular SAs were measured in the central 6-mm optical zone using wavefront aberrometry. Axial length (AL) and keratometry (K) were also evaluated in each eye. RESULTS: A total of 413 eyes in 234 patients were analyzed. The mean age of the patients was 66.8 ± 10.64 years. The mean AL and K values were 24.32 mm and 44.08 D, respectively. The mean corneal SA was 0.307 ± 0.135 µm and ocular SA was -0.042 ± 0.487 µm. Ocular and corneal SAs were significantly correlated (r2 = 0.04, p < 0.001). Corneal and ocular SAs were not significantly correlated with K (p = 0.096 and p = 0.634, respectively), but were significantly correlated with AL (p < 0.001). Multilinear regression showed that corneal SAs and age were the dependent variables that predicted ocular SAs (r2 = 0.143, F = 13.65, p < 0.01), especially in patients who were aged > 50 years, for whom a strongly significant positive correlation was found (r2 = 0.102, F = 11.10, p < 0.001). CONCLUSION: Corneal and ocular SAs varied among cataract patients and correlated with AL. After 50 years of age, ocular SAs increased significantly because of an increase in internal (lenticular) SAs. Corneal SAs in Taiwanese patients were larger than those in Japanese patients and similar to those in Chinese and Malaysian populations. Preoperative measurement of wavefront aberrations is necessary to select which aspherical intraocular lenses are most suitable for achieving better postoperative visual quality.