Validation of an Automated Early Treatment Diabetic Retinopathy Study Low-contrast Letter Acuity Test.

SIGNIFICANCE: Automated low-contrast letter acuity (LCLA) has several advantages: consistent luminance, reduced chance of individuals memorizing test letters, and convenient and accurate visual acuity reporting functions. Although automated LCLA might report slightly worse acuity than Sloan LCLA chart, considering its advantages, it may be a viable alternative to Sloan LCLA chart in clinical practice and research. PURPOSE: The purpose of this study was to determine the repeatability of an automated LCLA measurement and its agreement with the Sloan LCLA chart test in normal participants and reduced-vision participants. METHODS: Adult participants (n = 49) were measured with both automated Early Treatment Diabetic Retinopathy Study and Sloan LCLA tests, including normal and reduced-vision groups. Low-contrast letter acuity at two contrast levels (2.5 and 10%) was measured at 3 m in a random sequence with both LCLA tests. To test repeatability, participants were retested 1 week later. Repeatability of the two tests between two visits and agreement between automated and Sloan LCLA tests were evaluated using 95% limits of agreement. RESULTS: In terms of the 95% limits of agreement, the repeatability of both tests was as follows: automated LCLA at 2.5%, ±0.26; automated LCLA at 10%, ±0.22; Sloan LCLA at 2.5%, ±0.23, and Sloan LCLA at 10%, ±0.16. The agreement of the two tests was as follows: ±0.19 at 2.5% and ±0.24 at 10%. The automated LCLA at 2.5 and 10% levels was generally reported one-half to one logMAR line lower than Sloan LCLA (mean differences, -0.04 at 2.5% and -0.13 at 10%; paired t test, P < .05). CONCLUSIONS: The automated LCLA test shows fairly good test-retest repeatability at both 2.5 and 10% contrast levels. The agreement between the automated and the Sloan low-contrast letter acuity tests was comparable with test-retest agreement. Although the automated LCLA test reports slightly worse acuity than the Sloan LCLA test, it could be an appropriate alternative to the Sloan LCLA test.

Validation of an automated-ETDRS near and intermediate visual acuity measurement.

BACKGROUND: The aim of this study was to determine the repeatability of an automated-ETDRS (Early Treatment Diabetic Retinopathy Study) near and intermediate visual acuity measurement in subjects with normal visual acuity and subjects with reduced visual acuity. The agreement of automated-ETDRS with gold standard chart-based visual acuity measurement was also studied. METHODS: Fifty-one subjects were tested (aged 23 to 91 years; 33 subjects with normal visual acuity: 6/7.5 or better; 18 subjects with reduced visual acuity: 6/9 to 6/30). Near and intermediate visual acuity of one eye from each subject was measured with an automated tablet-computer system (M&S Technologies, Inc.) and Precision Vision paper chart in a random sequence. Subjects were retested one week later. Repeatability was evaluated using the 95 per cent limits of agreement (LoA) between the two visits. RESULTS: Average difference between automated-ETDRS near visual acuity and near visual acuity by paper chart was 0.02 ± 0.10 logMAR (p > 0.05). Agreement of near visual acuity between automated-ETDRS and paper chart was good, with 95 per cent LoA of ±0.19 logMAR. Furthermore, automated-ETDRS near visual acuity showed good repeatability (95 per cent LoA of ±0.20). Mean difference between automated-ETDRS intermediate visual acuity and intermediate visual acuity by paper chart was 0.02 ± 0.10 logMAR (p > 0.05). Agreement of intermediate visual acuity between automated-ETDRS and paper chart was good, with 95 per cent LoA of ±0.20 logMAR. In addition, automated-ETDRS intermediate visual acuity had good repeatability (95 per cent LoA of ±0.16). CONCLUSION: Automated-ETDRS near and intermediate visual acuity measurement showed good repeatability and agreement with the gold standard chart-based visual acuity measurement. The findings of this study indicate the automated visual acuity measurement system may have potential for use in both patient care and clinical trials.

Characterization of the phytochelatin synthase from the human parasitic nematode Ancylostoma ceylanicum.

Hookworm disease is a debilitating worm infection that affects hundreds of millions of people. Despite the existence of anthelmintic drugs, reports have testified of a decrease in efficacy of these drugs. Therefore, it is imperative to find new drugs and drug targets for hookworm disease treatment. In this study we identify the gene encoding the phytochelatin synthase in the human hookworm, Ancylostoma ceylanicum (AcePCS). Phytochelatin synthase catalyzes the production of metal chelating peptides, the phytochelatins, from glutathione (GSH). In plants, algae, and fungi phytochelatin production is important for metal tolerance and detoxification. Phytochelatin synthase proteins also function in the elimination of xenobiotics by processing GSH S-conjugates. We found that in vitro AcePCS could both synthesize phytochelatins and hydrolyze a GSH S-conjugate. Interestingly, the enzyme works through a thiol-dependent and, notably, metal-independent mechanism for both transpeptidase (phytochelatin synthesis) and peptidase (hydrolysis of GSH S-conjugates) activities. AcePCS mRNAs are expressed in vivo throughout the life cycle of A. ceylanicum. Mature adult male hookworms isolated from the small intestines of their hosts displayed significantly enhanced expression of AcePCS with transcript levels 5-fold greater than other developmental forms. Although the role of AcePCS in A. ceylanicum biology has yet to be fully investigated the results reported here provide encouraging evidence of the potential that this enzyme holds as a target for new chemotherapeutic intervention.

Towards an understanding of the function of the phytochelatin synthase of Schistosoma mansoni.

Phytochelatin synthase (PCS) is a protease-like enzyme that catalyzes the production of metal chelating peptides, the phytochelatins, from glutathione (GSH). In plants, algae, and fungi phytochelatin production is important for metal tolerance and detoxification. PCS proteins also function in xenobiotic metabolism by processing GSH S-conjugates. The aim of the present study is to elucidate the role of PCS in the parasitic worm Schistosoma mansoni. Recombinant S. mansoni PCS proteins expressed in bacteria could both synthesize phytochelatins and hydrolyze various GSH S-conjugates. We found that both the N-truncated protein and the N- and C-terminal truncated form of the enzyme (corresponding to only the catalytic domain) work through a thiol-dependant and, notably, metal-independent mechanism for both transpeptidase (phytochelatin synthesis) and peptidase (hydrolysis of GSH S-conjugates) activities. PCS transcript abundance was increased by metals and xenobiotics in cultured adult worms. In addition, these treatments were found to increase transcript abundance of other enzymes involved in GSH metabolism. Highest levels of PCS transcripts were identified in the esophageal gland of adult worms. Taken together, these results suggest that S. mansoni PCS participates in both metal homoeostasis and xenobiotic metabolism rather than metal detoxification as previously suggested and that the enzyme may be part of a global stress response in the worm. Because humans do not have PCS, this enzyme is of particular interest as a drug target for schistosomiasis.