Generation and characterization of avian single chain variable fragment against human Alpha-Enolase.

Overexpression of human alpha-enolase (hEno1)has been reported in a wide range of cancers and is tightly associated with poor prognosis, making it a remarkable biomarker and therapeutic target. In this study, polyclonal yolk-immunoglobulin (IgY) antibodies purified from hEno1-immunized chickens showed a noticeable specific humoral response. Phage display technology was used to construct two antibody libraries of IgY gene-derived single-chain variable fragments (scFvs) containing 7.8 × 107 and 5.4 × 107 transformants, respectively. Phage-based ELISA indicated that specific anti-hEno1 clones were significantly enriched. The nucleotide sequences of scFv-expressing clones were determined and classified into seven groups either in the short linker or the long linker. Moreover, higher mutation rates were revealed in the CDR regions, especially in the CDR3. Three distinguish antigenic epitopes were identified on the hEno1 protein. The binding activities of selected anti-hEno1 scFv on hEno1-positive PE089 lung cancer cells were confirmed using Western blot, flow cytometry, and immunofluorescence assay. In particular, hEnS7 and hEnS8 scFv antibodies significantly suppressed the growth and migration of PE089 cells. Taken together, these chicken-derived anti-hEno1 IgY and scFv antibodies have great potential to develop diagnostic and therapeutic agents for the treatment of lung cancer patients with high expression levels of hEno1 protein.

The Bottlenecks of Preparing Virus Particles by Size Exclusion for Antibody Generation.

Enterovirus 71 (EV71) is the major etiological agent contributing to the development of hand-foot-mouth disease (HFMD). There are not any global available vaccines or antibody drugs against EV71 released yet. In this study, we perform the virus immunization in a cost-effective and convenient approach by preparing virus particles from size exclusion and immunization of chicken. Polyclonal yolk-immunoglobulin (IgY) was simply purified from egg yolk and monoclonal single-chain variable fragments (scFv) were selected via phage display technology with two scFv libraries containing 6.0 × 106 and 1.3 × 107 transformants. Specific clones were enriched after 5 rounds of bio-panning and four identical genes were classified after the sequence analysis. Moreover, the higher mutation rates were revealed in the CDR regions, especially in the CDR3. IgY showed specific binding activities to both EV71-infected and Coxsackievirus 16-infected cell lysates and high infectivity inhibitory activity of EV71. However, while IgY detected a 37 kDa protein, the selected scFv seemingly detected higher size proteins which could be cell protein instead of EV71 proteins. Despite the highly effective chicken antibody generation, the purity of virus particles prepared by size exclusion is the limitation of this study, and further characterization should be carried out rigorously.

Flexible iris retractors for management of zonular dialysis during planned phacoemulsification.

BACKGROUND: The aim of this study was to determine the use of flexible iris retractors in four patients with zonular dialysis occurring during cataract surgery. METHODS: We describe the use of flexible iris retractors to stretch and fixate the capsulorhexis over the zonular dehiscence sector for patients with traumatic zonular disruption occurring during phacoemulsification, which stabilizes the capsular bag and facilitates cataract extraction without complications. RESULTS: Each of the four eyes was implanted with posterior chamber intraocular lens (PCIOL) successfully. The intraocular lenses (IOLs) remained well centered during the 12-month follow-up period and the patients enjoyed improved vision without complications. CONCLUSION: Flexible iris retractor is a simple, convenient and useful instrument for managing zonular dehiscence that occurred during cataract extraction. It facilitates the removal of the residual lens material and IOL implantation without major complications.

Pediatric rhegmatogenous retinal detachment in East Asians.

PURPOSE: To describe clinical features, predisposing factors, and surgical outcomes of retinal detachment in different age groups in a pediatric population. DESIGN: Retrospective, noncomparative, interventional case series. PARTICIPANTS: A total of 296 eyes from 278 children younger than 18 years of age underwent the first surgical procedure for rhegmatogenous retinal detachment between 1983 and 2003. They were divided into 3 age groups: 38 eyes in group 1 (0-10 years), 107 eyes in group 2 (11-15 years), and 151 eyes in group 3 (16-18 years). METHODS: Predisposing factors for retinal detachment were classified into 4 categories: (1) high myopia, (2) trauma, (3) congenital or developmental structural ocular abnormalities, and (4) previous intraocular surgery. MAIN OUTCOME MEASUREMENTS: Visual acuity and retinal reattachment. RESULTS: The mean age of the patients was 14.6 years; 73.7% were boys. The mean follow-up period was 51 months. At least 1 predisposing factor could be identified in 282 (95%) of study eyes. High myopia was most common in 111 eyes (38%). Thirty-one percent of eyes experienced trauma, 17% had structural abnormalities, and 5% underwent previous intraocular surgery. Macular involvement was found in 237 eyes (80%). The initial surgery was scleral buckling alone in 224 eyes (76%). Retinal reattachment was achieved in 214 eyes (72%) after the first operation and in 250 eyes (85%) at the end of intervention. Features seen in patients with a poor surgical outcome included congenital anomaly (P<0.001), previous intraocular surgery (P = 0.007), proliferative vitreoretinopathy grade C or worse (P<0.001), macula off (P = 0.001), total retinal detachment (P<0.001), and use of silicone oil (P<0.001). CONCLUSIONS: Myopia is one of the important predisposing factors of pediatric retinal detachment in East Asians. The predisposing factors, clinical features, timing of diagnosis, and frequency of proliferative vitreoretinopathy of retinal detachment are somewhat different in the 3 age groups considered. Because of higher proportions of congenital anomaly, total retinal detachment, proliferative vitreoretinopathy, and a low success rate with poor postoperative compliance, patients in group 1 experienced the lowest final reattachment rate and the worst visual recovery.

Retinopathy of prematurity: an evaluation in the Keelung area of Taiwan over a 10-year period.

BACKGROUND: We report on the experience of retinopathy of prematurity (ROP) at Chang Gang Memorial Hospital (CGMH), Keelung, Taiwan over a 10-year period. METHODS: A retrospective review was made of data of all premature infants with a diagnosis of ROP at Keelung CGMH between 1994 and 2003. Data on certain characteristics including gender, gestational age (GA), birth body weight (BBW), stage of ROP, and treatment modalities were collected and analyzed. RESULTS: Among the 458 infants screened for ROP, 148 eyes of 74 premature infants were diagnosed as having ROP. Threshold ROP occurred in 24 eyes of 12 infants. The average BBW and GA were significantly lower in the threshold than in the non-threshold ROP group (p < 0.05). According to medical records of the 24 eyes of the 12 patients receiving transscleral cryotherapy, anatomical success was attained in 13 (81.3%) of 16 eyes. CONCLUSIONS: This review found low GA and BBW to be major risk factors for ROP. This review shows that transscleral cryotherapy for treating threshold ROP achieved anatomical success in about 80% of eyes. However, myopia, amblyopia, and strabismus remain major sequelae. Laser therapy is now believed to be less damaging to ocular structures and just as effective as cryotherapy in treating ROP.