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Although the analysis of genetic interactions and networks is a powerful approach to understanding biology, it has not been applied widely to the pathogenic yeast Candida albicans Here, we describe the use of both screening and directed genetic interaction studies based on complex haploinsufficiency to probe the function of the R: egulation of A: ce2 and M: orphogenesis (RAM) pathway in C. albicans A library of 5200 Tn7-mutagenized derivatives of a parental strain heterozygous at CBK1, the key kinase in the RAM pathway, was screened for alterations in serum-induced filamentation. Following confirmation of phenotypes and identification of insertion sites by sequencing, a set of 36 unique double heterozygous strains showing complex haploinsufficiency was obtained. In addition to a large set of genes regulated by the RAM transcription factor Ace2, genes related to cell wall biosynthesis, cell cycle, polarity, oxidative stress, and nitrogen utilization were identified. Follow-up analysis led to the first demonstration that the RAM pathway is required for oxidative stress tolerance in a manner related to the two-component-regulated kinase Chk1 and revealed a potential direct connection between the RAM pathway and the essential Mps1 spindle pole-related kinase. In addition, genetic interactions with CDC42-related genes MSB1, a putative scaffold protein, and RGD3, a putative Rho GTPase-activating protein (GAP) were identified. We also provide evidence that Rgd3 is a GAP for Cdc42 and show that its localization and phosphorylation are dependent on Cbk1., (Copyright © 2016 by the Genetics Society of America.)

Purpose: To describe the morphological distribution of host tissue and microbial biofilms on the intraocular surfaces of Boston type I keratoprostheses (KPros) explanted because of corneal melt., Methods: Retrospective study of scanning electron microscopy (SEM) images from 4 explanted Boston type I KPros composed of polymethylmethacrylate and titanium. SEM images of KPro-associated ocular surfaces were reviewed for the presence of inflammatory cells, microbes, and/or biofilm formation. One sterile type I KPro was also imaged to serve as a (device only) control., Results: All 4 KPros were explanted because of culture-negative, clinically "sterile" donor corneal melt with impending KPro extrusion. In all cases, the rough, irregular surfaces of the device harbored more adherent corneal epithelium and stromacytes, inflammatory cells, and bacteria than the smooth, polished surface of the KPro optic. Two KPros showed not only evidence of prior bacterial colonization but marked biofilm formation., Conclusions: SEM images of explanted KPros explanted for "sterile" corneal melt demonstrated evidence of biofilm formation despite negative donor corneal cultures and the absence of clinical suspicion for infection. These results suggest that "sterile" corneal melt may be due to inflammatory host responses to low microbial burdens as seen in biofilms and/or released antigens after antibiotic-induced lysis. There was increased adherence of host tissue cells and microbial biofilms on the nonpolished surfaces of the KPro. Polishing the intraocular polymethylmethacrylate and titanium KPro surfaces may decrease microbial adhesion and biofilm formation in human subjects with KPros, but what impact this will have on rates of postoperative endophthalmitis is unknown.

Purpose: Tissues and biomaterials used for corneal surface repair require fulfilling specific optical standards prior to implantation in the patient. However, there is not a feasible evaluation method to be applied in clinical or Good Manufacturing Practice settings. In this study, we describe and assess an innovative easy-applied photographic-based method (PBM) for measuring functional optical blurring and transparency in corneal surface grafts., Methods: Plastic compressed collagen scaffolds (PCCS) and multilayered amniotic membranes (AM) samples were optically and histologically evaluated. Transparency and image blurring measures were obtained by PBM, analyzing photographic images of a standardized band pattern taken through the samples. These measures were compared and correlated to those obtained applying the Inverse Adding-Doubling (IAD) technique, which is the gold standard method., Results: All the samples used for optical evaluation by PBM or IAD were histological suitable. PCCS samples presented transmittance values higher than 60%, values that increased with increasing wavelength as determined by IAD. The PBM indicated that PCCS had a transparency ratio (TR) value of 80.3 ± 2.8%, with a blurring index (BI) of 50.6 ± 4.2%. TR and BI obtained from the PBM showed a high correlation (ρ>|0.6|) with the diffuse transmittance and the diffuse reflectance, both determined using the IAD (p<0.005). The AM optical properties showed that there was a largely linear relationship between the blurring and the number of amnion layers, with more layers producing greater blurring., Conclusions: This innovative proposed method represents an easy-applied technique for evaluating transparency and blurriness of tissues and biomaterials used for corneal surface repair.

Unlabelled: We describe a technique for image-guided femtosecond laser-assisted cataract surgery in a case of Peters anomaly type 2. The femtosecond laser technology enabled reliable construction of a complete capsulotomy despite central corneal opacification and a tented anterior capsule. Use of an image-guided femtosecond laser for cataract extraction in Peters anomaly type 2 is a safe method for removing these complex cataracts while preserving capsular and corneal integrity., Financial Disclosure: Dr. De la Cruz is a consultant to Alcon Surgical, Inc. No other author has a financial or proprietary interest in any material or method mentioned., (Copyright © 2015 ASCRS and ESCRS. Published by Elsevier Inc. All rights reserved.)

Drought severely limits forage productivity of C4 grasses across the tropics. The avoidance of water deficit by increasing the capacity for water uptake or by controlling water loss are common responses in forage C4 grasses. Napier grass (Pennisetum purpureum) and Brachiaria hybrid cv. Mulato II are tropical C4 grasses used for livestock production due to their reputed resistance to drought conditions. However, there is scant information on the mechanisms used by these grasses to overcome water-limited conditions. Therefore, assessments of cumulative transpired water, shoot growth, leaf rolling, leaf gas exchange, dry mass production and a number of morpho-physiological traits were recorded over a period of 21 days under well-watered or drought conditions. Drought reduced shoot dry mass of both grasses by 35 %, yet each grass exhibited contrasting strategies to cope with water shortage. Napier grass transpired most available water by the end of the drought treatment, whereas a significant amount of water was still available for Mulato II. Napier grass maintained carbon assimilation until the soil was fairly dry, whereas Mulato II restricted water loss by early stomatal closure at relatively wet soil conditions. Our results suggest that Napier grass exhibits a 'water-spending' behaviour that might be targeted to areas with intermittent drought stress, whereas Mulato II displays a 'water-saving' nature that could be directed to areas with longer dry periods., (Published by Oxford University Press on behalf of the Annals of Botany Company.)

Background: Being able to estimate (predict) the final spectrum of reflectance of a biomaterial, especially when the final color and appearance are fundamental for their clinical success (as is the case of dental resin composites), could be a very useful tool for the industrial development of these type of materials. The main objective of this study was the development of predictive models which enable the determination of the reflectance spectrum of experimental dental resin composites based on type and quantity of pigments used in their chemical formulation., Methods: 49 types of experimental dental resin composites were formulated as a mixture of organic matrix, inorganic filler, photo activator and other components in minor quantities (accelerator, inhibitor, fluorescent agent and 4 types of pigments). Spectral reflectance of all samples were measured, before and after artificial chromatic aging, using a spectroradiometer. A Multiple Nonlinear Regression Model (MNLR) was used to predict the values of the Reflectance Factors values in the visible range (380 nm-780 nm), before and after aging, from % Pigment (%P1, %P2, %P3 and %P4) within the formulation., Results: The average value of the prediction error of the model was 3.46% (SD: 1.82) across all wavelengths for samples before aging and 3.54% (SD: 1.17) for samples after aging. The differences found between the predicted and measured values of the chromatic coordinates are smaller than the acceptability threshold and, in some cases, are even below the perceptibility threshold., Conclusions: Within the framework of this pilot study, the nonlinear predictive models developed allow the prediction, with a high degree of accuracy, of the reflectance spectrum of the experimental dental resin composites.