n 1619, Scheiner provided the first precise description of the corneal shape using glass balls of known curvatures.1 From that first description, many other diagnostic tools have been developed to describe corneal shape, from keratometry to corneal topography (front surface curvature maps), 2 then into three-dimensional corneal tomography systems.3 More recently, it has been shown that corneal biomechanical behavior plays an important role in maintaining corneal shape, which is necessary for light refraction and clear vision, 4 and should therefore be considered in understanding the development of ectatic diseases 5,6 and the results of surgery. 4,7 Until recently, the evaluation of corneal biomechanical properties had been restricted to ex vivo laboratory studies 5,8 and mathematical corneal models. METHODS: Seven hundred five healthy patients were included in this multicenter retrospective study. The biomechanical response data were analyzed to obtain normative values with their dependence on corrected and clinically validated intraocular pressure estimates developed using the finite element method (bIOP), central corneal thickness (CCT), and age, and to evaluate the influence of bIOP, CCT, and age. RESULTS:The results showed that all DCRs were correlated with bIOP except deflection amplitude (DefA) ratio, highest concavity (HC) radius, and inverse concave radius. The analysis of the relationship of DCRs with CCT indicated that HC radius, inverse concave radius, deformation amplitude (DA) ratio, and DefA ratio were correlated with CCT (rho values of 0.343, -0.407, -0.444, and -0.406, respectively). The age group subanalysis revealed that primarily whole eye movement followed by DA ratio and inverse concave radius were the parameters that were most influenced by age. Finally, custom software was created to compare normative values to imported examinations.CONCLUSIONS: HC radius, inverse concave radius, DA ratio, and DefA ratio were shown to be suitable parameters to evaluate in vivo corneal biomechanics due to their independence from IOP and their correlation with pachymetry and age. The creation of normative values allows the interpretation of an abnormal examination without the need to match every case with another normal patient matched for CCT and IOP.[J Refract Surg. 2016;32(8):550-561.]

Exosomes, small membrane vesicles secreted by a multitude of cell types, are involved in a wide range of physiological roles such as intercellular communication, membrane exchange between cells, and degradation as an alternative to lysosomes. Because of the small size of exosomes (30-100 nm) and the limitations of common separation procedures including ultracentrifugation and flow cytometry, size-based fractionation of exosomes has been challenging. In this study, we used flow field-flow fractionation (FlFFF) to fractionate exosomes according to differences in hydrodynamic diameter. The exosome fractions collected from FlFFF runs were examined by transmission electron microscopy (TEM) to morphologically confirm their identification as exosomes. Exosomal lysates of each fraction were digested and analyzed using nanoflow LC-ESI-MS-MS for protein identification. FIFFF, coupled with mass spectrometry, allows nanoscale size-based fractionation of exosomes and is more applicable to primary cells and stem cells since it requires much less starting material than conventional gel-based separation, in-gel digestion and the MS-MS method.

Insulin and juvenile hormone signaling direct entry of the mosquito Culex pipiens into its overwintering adult diapause, and these two critical signaling pathways appear to do so by converging on the regulation of forkhead transcription factor (FOXO). Diapause is a complex phenotype, and FOXO emerges as a prime candidate for activating many of the diverse physiological pathways that generate the diapause phenotype. Here, we used ChIP sequencing to identify direct targets of FOXO. The nearest gene in a 10-kb region surrounding a predicted binding site was extracted for each binding site, resulting in a dataset containing genes potentially regulated by FOXO. By selecting candidate genes based on their functional relevance to diapause, we identified five gene categories of potential interest, including stress tolerance, metabolic pathways, lifespan extension, cell cycle and growth regulation, and circadian rhythms. Twelve targets were prioritized for further analysis, 10 of which were validated by ChIP-quantitative PCR and quantitative real-time PCR. These 10 genes activated by FOXO are highly up-regulated during diapause and are thus strong candidates for implementation of the diapause syndrome.diapause | Culex pipiens | forkhead transcription factor | insulin D iapause is a complex phenotype characterized not only by its hallmark trait of arrested development, but also by switches in behavior, suppressed metabolism, and enhanced responses to a range of environmental insults (1, 2). In the mosquito Culex pipiens, the diapause syndrome encompasses a cessation of reproductive maturation, a switch from blood feeding to sugar feeding, migration to protective overwintering sites, suppression of metabolism, and enhanced stress responses to combat low temperature, desiccation, and pathogens (3-7). These characteristics are indeed diverse, and it is not at all clear how the major hormonal signals that govern diapause are translated into the suite of features that comprise the diapause phenotype. A shutdown in the production of juvenile hormone (JH) is a nearly universal endocrine mechanism for insect diapauses that occur in the adult stage (8), and, more recently, the insulin signaling (IS) pathway has also been implicated in diapause regulation (9-12). The JH and IS pathways merge, as shown in Fig. 1. The IS pathway is linked to stimulation of the corpora allata to produce JH, and the IS and JH signaling pathways appear to collaboratively mediate expression of the forkhead transcription factor (FOXO). In the absence of IS and JH signaling, FOXO is activated, and this activation is critical for fat accumulation (13), cessation of ovarian development (9), and the elevated antioxidant responses (14) associated with diapause in Cx. pipiens. The genetic pathway involving IS and FOXO appears to also play a critical role in regulating dauer formation in the nematode Caenorhabditis elegans (15) and reproductive diapause in the fruit fly Drosophila melanogaster (16), thus suggesting a common mechanism regulating these diverse for...

Commensal microbiota are well known to play an important role in antiviral immunity by providing immune inductive signals; however, the consequence of dysbiosis on antiviral immunity remains unclear. We demonstrate that dysbiosis caused by oral antibiotic treatment directly impairs antiviral immunity following viral infection of the vaginal mucosa. Antibiotic-treated mice succumbed to mucosal herpes simplex virus type 2 infection more rapidly than water-fed mice, and also showed delayed viral clearance at the site of infection. However, innate immune responses, including type I IFN and proinflammatory cytokine production at infection sites, as well as induction of virusspecific CD4 and CD8 T-cell responses in draining lymph nodes, were not impaired in antibiotic-treated mice. By screening the factors controlling antiviral immunity, we found that IL-33, an alarmin released in response to tissue damage, was secreted from vaginal epithelium after the depletion of commensal microbiota. This cytokine suppresses local antiviral immunity by blocking the migration of effector T cells to the vaginal tissue, thereby inhibiting the production of IFN-γ, a critical cytokine for antiviral defense, at local infection sites. These findings provide insight into the mechanisms of homeostasis maintained by commensal bacteria, and reveal a deleterious consequence of dysbiosis in antiviral immune defense.commensal microbiota | dysbiosis | IL-33 | herpes simplex virus type 2 | genital tract

The dynamic Scheimpflug analyzer showed stable biomechanically corrected IOP measurement before and after surgery. The changes in dynamic corneal response parameters were smaller with transepithelial PRK than with femtosecond-assisted LASIK, indicating less of a biomechanical effect with transepithelial PRK.