The relationship between myopia and near work, time outdoors and socioeconomic status in children and adolescents.

BACKGROUND: To investigate environmental and social risk factors for myopia in children and adolescents in Germany. METHODS: 1437 children aged between 3 and 18 inclusive were examined as part of the LIFE Child study based in Leipzig, Germany. Information about leisure time activities and social status was ascertained by parents and children in a questionnaire. Refractive status was attained by measuring noncycloplegic autorefraction. Myopia was defined as spherical equivalent (SE) ≤ - 0.75 D. Risk factors were identified using multiple logistic regression analysis. RESULTS: In multiple logistic regression analysis, myopia was significantly associated with less frequent outdoor activity ("once a week" vs. "twice a week or more": odds ratio (OR) 4.35, 95% confidence interval (CI) 1.89-9.98, p<0.01) and longer near work sessions (1-2 h vs. < 1 h: OR 1.83, CI 1.10-3.04, p=0.02; > 3 h vs. < 1 h: OR 3.71, CI 1.43-9.61, p<0.01) after adjustment for age, sex and socioeconomic status (SES). Duration of outdoor activity, near work frequency and SES showed no significant association with myopia (p > 0.05). Children with a lower SES were involved in longer periods of outdoor and near work activities but on fewer occasions over the course of the week, although this connection was not significant. CONCLUSION: Myopia is associated with environmental factors. The present findings suggest that daily exposure to sunlight and a restriction of long-duration near work activities might protect against pathological eye growth. Prevention strategies should be implemented for children at all ages.

Longitudinal analysis of axial length growth in a German cohort of healthy children and adolescents.

PURPOSE: To generate continuous growth curves for axial length (AL) in German children. We hypothesise that percentile curves of AL can be used as a predictive measure of myopia. METHODS: In this longitudinal and cross-sectional LIFE Child Study, children's non-cycloplegic refraction data was collected using the Zeiss i.Profiler plus while AL was measured using the Haag-Streit Lenstar. Reference growth curves were estimated as a continuous non-parametric function of age. RESULTS: Data from 4511 visits of 1965 participants (1021 boys and 944 girls) between 3 and 18 years of age were analysed. For all ages and percentiles, the estimated AL was higher in boys than girls. AL differences between boys and girls were most pronounced in the 98th percentile at 3 years of age, being 0.93 mm longer eyes in boys. This difference decreased to 0.21 mm at 18 years of age. While the lower percentiles of AL reach their final value around age 13, the 50th percentile was still increasing by 0.05 mm per year until the end of the observation period. While, in general, children with longer eyes are more likely to develop myopia, this relationship is weaker between the ages of 5 and 8. CONCLUSION: The LIFE Child Study data provides European AL data. In both Germany and China, AL has comparable growth rates when the baseline ALs are compared as percentiles. Thus, percentile curves of AL can be used as a predictive measure for the likelihood of developing as well as the progression of myopia.

Impaired visual acuity caused by uncorrected refractive errors and amblyopia in a German paediatric cohort.

PURPOSE: This study describes the prevalence of visual impairment caused by uncorrected distance refractive error and the spectacle coverage in a large cohort of German children and adolescents. Furthermore, we determined the prevalence of amblyopia and amblyopia treatment. METHODS: Optometrists carried out visual acuity (VA) testing, non-cycloplegic autorefraction, VA retesting with the refractive correction obtained by autorefraction, and if necessary subjective refraction and cover-test in 1,874 subjects (901 girls/973 boys), aged 3 to 16 years. Additionally, a questionnaire on the history of previous eye care was completed. RESULTS: Distance visual impairment (VI) with VA worse than 6/18 Snellen in the better eye was present in 0.5% (95% confidence interval [95% CI]: 0.2% to 0.9%) of children. Logistic regression showed a significant positive association between VI and older age (OR = 1.21, P = 0.03). Gender differences were not found (P = 0.77). The spectacle coverage of all participants was 74.8% (95% CI: 65.2% to 82.8%). Amblyopia was found in 29 participants (1.5%; 95% CI: 1.0% to 2.2%) with no age or sex differences. The causes of unilateral amblyopia (27 cases, 93.1%) were anisometropia in 55.6% of children, strabismus in 25.9% and the combination of these factors in 18.5%. All children with bilateral amblyopia (2 cases, 6.9%) showed bilateral high ametropia. We observed 15 children with current amblyopia, who had been treated with occlusion therapy in the past (62.2%; 95% CI: 42.7% to 83.6%). CONCLUSIONS: The prevalence of impaired VA caused by uncorrected refractive error and amblyopia is comparable with previous studies conducted in urban areas worldwide. Adding the measurement of refractive error to existing medical check-ups, possibly using autorefraction, would be desirable.

Reference curves for refraction in a German cohort of healthy children and adolescents.

PURPOSE: Percentile curves of refractive development for German children were generated. We hypothesize that refraction in children in central Europe might differ from data in central Asia. METHODS: Non-cycloplegic refraction was measured using the ZEISS i.Profiler plus (Carl Zeiss Vision GmbH, Germany) in 1999 children, of which were 1046 male and 953 female, aged 3 to 18 years. Reference curves were calculated with the R-package GAMLSS as continuous function of age. RESULTS: There were only little differences for all centiles between the genders at 3 years and a general trend towards more myopia with increasing age. For the 97th centile and the 3rd centile, girls showed higher myopia/ less hyperopia than boys. Between the age of 3 and 18, the median refraction became -0.68 D and -0.74 D more myopic for boys and girls, respectively. At the same time, the 97th centile for boys changed +0.29 D towards hyperopia and in girls -0.52 D towards myopia. A general myopic trend was seen in the 3rd centile, which was -2.46 D for boys and -2.98 D for girls. For both genders, the median became less than zero at the age of 10 years but did not become myopic (less than -0.5 D) up to the age of 18. CONCLUSION: Our analysis presents the first reference curve for refraction in central Europe. In comparison to data from China and Korea, there is only little difference at the age of 5 years in all centiles which then increases continuously. For all ethnicities, a trend towards myopia with increasing age could be observed, but myopia progression is much higher in China and Korea than in Germany. The most marked differences can be seen in the lower centiles. Further investigations should clarify whether commencement of preschool activities with prolonged near-work initiates the divergence in refractive development.

Complete Sequence of Probiotic Symbioflor 2 Escherichia coli Strain G3/10 and Draft Sequences of Symbioflor 2 E. coli Strains G1/2, G4/9, G5, G6/7, and G8.

The complete genome of probiotic Escherichia coli strain G3/10 is presented here. In addition, the probiotic E. coli strains G1/2, G4/9, G5, G6/7, and G8 are presented in draft form. These six strains together comprise the probiotic product Symbioflor 2 (DSM 17252).

Secretory expression of biologically active human Herpes virus interleukin-10 analogues in Escherichia coli via a modified Sec-dependent transporter construct.

BACKGROUND: Interleukin-10 homologues encoded by Herpes viruses such as Epstein-Barr virus (EBV) and human cytomegalovirus (HCMV) hold interesting structural and biological characteristics compared to human interleukin-10 (hIL-10) that render these proteins promising candidates for therapeutic application in inflammatory bowel disease (IBD). Intestinal delivery of cytokines using bacterial carriers as chassis represents a novel approach for treatment of IBD patients. For proof of concept, a Sec-dependent transporter construct was designed for secretory expression of recombinant viral IL-10 proteins in the periplasm of Escherichia coli laboratory strain BL21 (DE3), which might serve as part of a prospective lysis based delivery and containment system. RESULTS: The signal peptide of E. coli outer membrane protein F fused to the mature form of the viral IL-10 proteins enabled successful transport into the periplasm, a compartment which seems crucial for proper assembly of the dimeric configuration of the cytokines. Cytokine concentrations in different bacterial compartments were determined by ELISA and achieved yields of 67.8 ng/ml ± 24.9 ng/ml for HCMV IL-10 and 1.5 µg/ml ± 841.4 ng/ml for EBV IL-10 in the periplasm. Immunoblot analysis was used to confirm the correct size of the E. coli-derived recombinant cytokines. Phosphorylation of signal transducer and activator of transcription 3 (STAT3) as part of the signal transduction cascade after IL-10 receptor interaction, as well as suppression of tumor necrosis factor α (TNF-α) release of lipopolysaccharide-stimulated mouse macrophages were used as read-out assays for proving in vitro biological activity of the E. coli derived, recombinant viral IL-10 counterparts. CONCLUSIONS: In this study, proof of principle is provided that E. coli cells are a suitable chassis for secretory expression of viral IL-10 cytokines encoded by codon-optimized synthetic genes fused to the E. coli ompF signal sequence. In vitro biological activity evidenced by activation of transcription factor STAT3 and suppression of TNF-α in mammalian cell lines was shown to be strictly dependent on export of viral IL-10 proteins into the periplasmic compartment. E. coli might serve as carrier system for in situ delivery of therapeutic molecules in the gut, thus representing a further step in the development of novel approaches for treatment of IBD.

Improving health from the inside: Use of engineered intestinal microorganisms as in situ cytokine delivery system.

The anti-inflammatory cytokine interleukin-10 and its viral homologs were chosen as model proteins for the development of drug delivery systems based on probiotic carriers like E. coli Nissle 1917, E. coli G3/10, and Saccharomyces boulardii. Exterior cytokine secretion was achieved by a modified E. coli hemolysin transporter. Release of interleukin-10 transported to the periplasm via the OmpF signal peptide was enabled by a T4 phage lysis system under control of the araC PBAD activator-promoter. The yield of interleukin-10 delivered by the phage lysis system was too low for functional analysis whereas the fusion protein secreted by the hemolysin transporter proved to be biologically inactive. Moreover, partial processing of the fusion protein by the E. coli membrane protease OmpT had no effect on the protein's functionality. Using the α-mating factor signal sequence, the yeast S. boulardii proved to be suitable for secretory expression of biologically active viral interleukin-10.

Periplasmic delivery of biologically active human interleukin-10 in Escherichia coli via a sec-dependent signal peptide.

Interleukin-10 (IL-10) is a potent anti-inflammatory cytokine, with therapeutic applications in inflammatory bowel disease. For the in situ delivery of IL-10 by Escherichia coli as carrier chassis, a modified transporter was designed with the ability to secrete biologically active IL-10. De novo DNA synthesis comprised a 561-bp fragment encoding the signal sequence of the E. coli outer membrane protein F fused in frame to an E. coli codon-optimized mature human IL-10 gene under control of a T7 promoter. The construct was overexpressed in E. coli laboratory strains, E. coli BL21 (DE3) and E. coli MDS42:T7. The mean concentrations of human IL-10 in the periplasm and culture supernatant of E. coli BL21 (DE3) were 355.8 ± 86.3 and 5.7 ± 1.7 ng/ml, respectively. The molecular mass of the recombinant E. coli-derived human IL-10 was 19 kDa, while under non-reducing conditions the native IL-10 dimer could be demonstrated. Reduction of tumor necrosis factor-α secretion in lipopolysaccharide-stimulated mouse macrophages and detection of the activated form of the transcription factor signal transducer and activator of transcription protein 3 proved the biological activity of the bacteria-produced human IL-10.