Soluble prefusion-closed HIV-envelope trimers with glycan-covered bases.

Soluble HIV-1-envelope (Env) trimers elicit immune responses that target their solvent-exposed protein bases, the result of removing these trimers from their native membrane-bound context. To assess whether glycosylation could limit these base responses, we introduced sequons encoding potential N-linked glycosylation sites (PNGSs) into base-proximal regions. Expression and antigenic analyses indicated trimers bearing six-introduced PNGSs to have reduced base recognition. Cryo-EM analysis revealed trimers with introduced PNGSs to be prone to disassembly and introduced PNGS to be disordered. Protein-base and glycan-base trimers induced reciprocally symmetric ELISA responses, in which only a small fraction of the antibody response to glycan-base trimers recognized protein-base trimers and vice versa. EM polyclonal epitope mapping revealed glycan-base trimers -even those that were stable biochemically- to elicit antibodies that recognized disassembled trimers. Introduced glycans can thus mask the protein base but their introduction may yield neo-epitopes that dominate the immune response.

Acquisition and annotation in high resolution in vivo digital biopsy by confocal microscopy for diagnosis in oral precancer and cancer.

Introduction: Scanned fibre endomicroscopes are full point-scanning confocal microscopes with submicron lateral resolution with an optical slice thickness thin enough to isolate individual cell layers, allow active positioning of the optical slice in the z-axis and collection of megapixel images. Here we present descriptive findings and a brief atlas of an acquisition and annotation protocol high resolution in vivo capture of oral mucosal pathology including oral squamous cell carcinoma and dysplasia using a fluorescence scanned fibre endomicroscope with 3 topical fluorescent imaging agents: fluorescein, acriflavine and PARPi-FL. Methods: Digital biopsy was successfully performed via an acquisition protocol in seventy-one patients presenting for investigation of oral mucosal abnormalities using a miniaturized, handheld scanned fibre endoscope. Multiple imaging agents were utilized and multiple time points sampled. Fifty-nine patients had a matched histopathology correlating in location with imaging. The images were annotated back to macrographic location using a purpose-built software, MouthMap™. Results: Acquisition and annotation of cellular level resolved images was demonstrated with all 3 topical agents. Descriptive observations between clinically or histologically normal oral mucosa showed regular intranuclear distance, a regular nuclear profile and fluorescent homogeneity. This was dependent on the intraoral location and type of epithelium being observed. Key features of malignancy were a loss of intranuclear distance, disordered nuclear clustering and irregular nuclear fluorescence intensity and size. Perinuclear fluorescent granules were seen in the absence of irregular nuclear features in lichenoid inflammation. Discussion: High resolution oral biopsy allows for painless and rapid capture of multiple mucosal sites, resulting in more data points to increase diagnostic precision. High resolution digital micrographs can be easily compared serially across multiple time points utilizing an annotation software. In the present study we have demonstrated realization of a high-resolution digital biopsy protocol of the oral mucosa for utility in the diagnosis of oral cancer and precancer..

Confocal microscopy in oral cancer and oral potentially malignant disorders: A systematic review.

OBJECTIVE: To systematically identify and summarise current research on the utility of confocal microscopy in oral squamous cell carcinoma and oral epithelial dysplasia in oral potentially malignant disorders. METHODS: Databases Medline, Embase, Evidence-Based Medicine, and Web of Science were searched with articles screened and included if their primary objective was the use of a confocal microscope in diagnosis of oral cancer or epithelial dysplasia, in vivo or ex vivo. RESULTS AND DISCUSSION: Twenty-eight relevant studies were identified of which 21 studies included oral squamous cell carcinoma specimens. Fifteen studies included in vivo use. The studies included both qualitative and fluorescence confocal microscope and reflectance confocal microscope analysis along with quantitative analysis of carcinoma and dysplasia. Thirteen studies reported the predictive value of their confocal device in the diagnosis of dysplasia and carcinoma. The quantitative software-based studies show promise in objectifying the diagnostic process for identifying abnormalities within the microstructure of the oral mucosa. CONCLUSIONS: There was heterogeneity in the criteria for diagnosis of dysplasia and oral squamous cell carcinoma with experience levels of assessors impacting method efficacy. Both qualitative and quantitative confocal assessment methodologies have been explored, the latter highlighting the potential of future machine-augmented diagnostic precision.

Kaempferol Protects Against Retinal Photoreceptor Degeneration in a Mouse Model of Light-Induced Retinal Injury.

Purpose: Age-related macular degeneration (AMD) is a leading cause of blindness in developed countries with little in the way of treatment that prevents progression to end-stage disease. Kaempferol (KF) is a plant-derived dietary flavonoid that has demonstrated as a strong antioxidant showing neuroprotection in stroke models. We hypothesize that KF has protective effects against retinal degeneration and may serve as a therapeutic agent against AMD. Methods: BALB/c albino mice were assigned to 1 of 2 groups: control-treated or KF-treated retinal light injury mice. Mice were exposed to 8,000 lux cool white fluorescent light for 2 h to induce light injury. Control or KF was injected intraperitoneally after light injury for 5 days. Scotopic electroretinography (ERG) was recorded before light injury and 7 days after light injury. The retinal morphology and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assays were performed after light injury. Results: ERG a- and b-wave amplitudes were significantly reduced in the retinal light injury group compared with the nonretinal light injury group. Retinal light injury produced markedly thinning of the outer nuclear layer along with significant TUNEL-positive signals. In contrast KF treatments significantly attenuated reduction of ERG a- and b- wave amplitudes and the loss of the outer nuclear layer. Conclusions: KF protects retinal photoreceptors and preserves retinal function against retinal degeneration caused by light injury. These initial findings suggest that KF may represent a novel therapy for retinal degenerative conditions such as AMD.

Bio-energy generation in an affordable, single-chamber microbial fuel cell integrated with adsorption hybrid system: effects of temperature and comparison study.

A microbial fuel cell (MFC) integrated with adsorption system (MFC-AHS) is tested under various operating temperatures with palm oil mill effluent as the substrate. The optimum operating temperature for such system is found to be at ∼35°C with current, power density, internal resistance (Rin), Coulombic efficiency (CE) and maximum chemical oxygen demand (COD) removal of 2.51 ± 0.2 mA, 74 ± 6 mW m-3, 25.4 Ω, 10.65 ± 0.5% and 93.57 ± 1.2%, respectively. Maximum current density increases linearly with temperature at a rate of 0.1772 mA m-2 °C-1, whereas maximum power density was in a polynomial function. The temperature coefficient (Q10) is found to be 1.20 between 15°C and 35°C. Present studies have demonstrated better CE performance when compared to other MFC-AHSs. Generally, MFC-AHS has demonstrated higher COD removals when compared to standalone MFC regardless of operating temperatures. ABBREVIATIONS: ACFF: activated carbon fiber felt; APHA: American Public Health Association; CE: Coulombic efficiency; COD: chemical oxygen demand; ECG: electrocardiogram; GAC: granular activated carbon; GFB: graphite fiber brush; MFC: microbial fuel cell; MFC-AHS: microbial fuel cell integrated with adsorption hybrid system; MFC-GG: microbial fuel cell integrated with graphite granules; POME: palm oil mill effluent; PTFE: polytetrafluoroethylene; SEM: scanning electron microscope.

Performance evaluation of a hybrid system for efficient palm oil mill effluent treatment via an air-cathode, tubular upflow microbial fuel cell coupled with a granular activated carbon adsorption.

An air-cathode MFC-adsorption hybrid system, made from earthen pot was designed and tested for simultaneous wastewater treatment and energy recovery. Such design had demonstrated superior characteristics of low internal resistance (29.3Ω) and favor to low-cost, efficient wastewater treatment and power generation (55mW/m(3)) with average current of 2.13±0.4mA. The performance between MFC-adsorption hybrid system was compared to the standalone adsorption system and results had demonstrated great pollutants removals of the integrated system especially for chemical oxygen demand (COD), biochemical oxygen demand (BOD3), total organic carbon (TOC), total volatile solids (TVS), ammoniacal nitrogen (NH3-N) and total nitrogen (TN) because such system combines the advantages of each individual unit. Besides the typical biological and electrochemical processes that happened in an MFC system, an additional physicochemical process from the activated carbon took place simultaneously in the MFC-adsorption hybrid system which would further improved on the wastewater quality.

Observational constraints on mixed-phase clouds imply higher climate sensitivity.

Global climate model (GCM) estimates of the equilibrium global mean surface temperature response to a doubling of atmospheric CO2, measured by the equilibrium climate sensitivity (ECS), range from 2.0° to 4.6°C. Clouds are among the leading causes of this uncertainty. Here we show that the ECS can be up to 1.3°C higher in simulations where mixed-phase clouds consisting of ice crystals and supercooled liquid droplets are constrained by global satellite observations. The higher ECS estimates are directly linked to a weakened cloud-phase feedback arising from a decreased cloud glaciation rate in a warmer climate. We point out the need for realistic representations of the supercooled liquid fraction in mixed-phase clouds in GCMs, given the sensitivity of the ECS to the cloud-phase feedback.

Continuous asymmetric ketone reduction processes with recombinant Escherichia coli.

The reduction of methyl acetoacetate was carried out in continuously operated biotransformation processes catalyzed by recombinant Escherichia coli cells expressing an alcohol dehydrogenase from Lactobacillus brevis. Three different cell types were applied as biocatalysts in three different cofactor regeneration approaches. Both processes with enzyme-coupled cofactor regeneration catalyzed by formate dehydrogenase or glucose dehydrogenase are characterized by a rapid deactivation of the biocatalyst. By contrast the processes with substrate-coupled cofactor regeneration by alcohol dehydrogenase catalyzed oxidation of 2-propanol could be run over a period of 7 weeks with exceedingly high substrate and cosubstrate concentrations of up to 2.5 and 2.8 mol L(-1), respectively. Even under these extreme conditions, the applied biocatalyst showed a good stability with only marginal leakage of intracellular cofactors.