Miracle-dz/UniFAR-FTUs huggingface dataset:
Effect of coupling on volume-regulatory response of ciliary epithelial cells suggests mechanism for secretion.

The ciliary epithelium of the eye secretes the aqueous humor. It is a double epithelium arranged so that the apical surfaces of the nonpigmented ciliary epithelial (NPCE) and pigmented ciliary epithelial (PCE) cells face each other and the basolateral membranes face the inside of the eye and the blood, respectively. We have investigated the volume responses of both single cells and coupled pairs from this tissue to osmotic challenge. Both NPCE and PCE cells undergo regulatory volume increase (RVI) and decrease (RVD) when exposed to hyper- and hyposmotic solution, respectively. In hyposmotic solution single cells swell and return to their original volumes within ∼3 min. In nonpigmented cells RVD could be inhibited by blockers of volume-activated Cl-channels [tamoxifen (100%) > quinidine (87%) > DIDS (84%) > 5-nitro-2-(3-phenylpropylamino)benzoic acid (80%) > SITS (58%)] and K+ channels [Ba2+(31%)]. However, in PCE cells these inhibitors and additionally tetraethylammonium and Gd3+ were without effect. Only bumetanide, an inhibitor of Na+-K+-2Cl-cotransport, was found to have any effect on RVD in PCE cells. NPCE-PCE cell coupled pairs also underwent RVD, but with altered kinetics. The onset of RVD of the PCE cell in a pair occurred ≈80 s before that of the NPCE cell, and the peak swell was reduced. This is consistent with fluid movement from the PCE to the NPCE cell. The effect of the volume-activated Cl- channel inhibitor tamoxifen was to eliminate this difference in the times of onset of RVD in coupled cell pairs and to inhibit RVD in both the NPCE and PCE cells partially. On the basis of these observations we suggest that fluid is transferred from the PCE to the NPCE cell in coupled pairs during cell swelling and the subsequent RVD. Furthermore, we speculate that reciprocal RVI-RVD could underlie aqueous humor secretion.
Development of optical fibre chemical probes by oblique angle deposition

An optical fibre chemical sensor based on the technique of surface-enhanced Raman scattering (SERS) has been developed. SERS relies on the close interaction between the target analyte and a nanostructured metal surface. This nanostructure was fabricated using the method of oblique angle deposition (OAD) under thermal evaporation. The deposition was carried out by holding the sample surface at an oblique angle to the metal vapour flux, which results in a columnar morphology of the thin film. The viability of thermal OAD for SERS substrates was initially determined by the use of a planar silicon wafer with a thermal oxide layer. It was found that a vapour deposition angle of 86○ provided the optimum nanostructure for SERS performance, using thiophenol (C6H5SH) as a test analyte. Strict control of the evaporation current was required to minimise radiant heating inside the evaporation chamber, as any increase in the temperature resulted in a high variability in the SERS response across different deposition runs. This appeared to be due to increased adatom diffusivity on the nanorod surface triggered by the thermal energy, which in turn resulted in morphological changes of the nanostructure. Controlling the heating by regulating the evaporation current produced a highly reproducible SERS substrate which exhibited a repeatability of ≲10% based on relative standard deviation (RSD). This insight was used to extend the technique to the tip of a standard multi-mode optical fibre. The earlier results were verified by further tests, and the SERS activity was found to increase with film thickness, with a maximum activity at a nanorod length of 260 ±28 nm. Although the OAD nanorod structures fabricated on the fibre tip exhibited excellent SERS activity in direct excitation, the signal collection efficiency was found to be reduced down to between 8% and 1%, in the remote excitation where the substrate was excited by coupling light through the fibre. The main reason for this discrepancy was revealed to be due to an incomplete overlap between the confocal light collection area and the spatial distribution of the scattered light in the fibre core. Other factors such
Glucose transporters control gene expression of aldose reductase, PKCα, and GLUT1 in mesangial cells in vitro.

The process linking increased glucose utilization and activation of metabolic pathways leading to end-organ damage from diabetes is not known. We have previously described rat mesangial cells that were transduced to constitutively express the facilitative glucose transporter 1 (GLUT1, MCGT1 cells) or bacterial β-galactosidase (MCLacZ, control cells). Glucose transport was rate limiting for extracellular matrix production in the MCGT1 cells. In the present work, we investigated the effect of GLUT1 overexpression in mesangial cells on aldose reductase (AR), protein kinase Cα (PKCα), and native GLUT1 transcript levels, to determine whether changes in GLUT1 alone could regulate their expression in the absence of high extracellular glucose concentrations. MCGT1 cells grown in normal (8 mM) or elevated (20 mM) glucose had elevated abundance of AR, PKCα, and the native GLUT1 transcripts compared with control cells. AR protein levels, AR activity, sorbitol production, and PKCα protein content were also greater in the MCGT1 cells than in control cells grown in the same media. This is the first report of the concomitant activation of AR, PKCα, and GLUT1 genes by enhanced GLUT1 expression. We conclude that increased GLUT1 expression leads to a positive feedback of greater GLUT1 expression, increased AR expression and activity with polyol accumulation, and increased total and active PKCα protein levels, which leads to detrimental stimulation of matrix protein synthesis by diabetic mesangial cells.
Muscle net glucose uptake and glucose kinetics after endurance training in men.

We evaluated the hypotheses that alterations in glucose disposal rate (Rd) due to endurance training are the result of changed net glucose uptake by active muscle and that blood glucose is shunted to working muscle during exercise requiring high relative power output. We studied leg net glucose uptake during 1 h of cycle ergometry at two intensities before training [45 and 65% of peak rate of oxygen consumption (V˙o 2 peak)] and after training [65% pretrainingV˙o 2 peak, same absolute workload (ABT), and 65% posttrainingV˙o 2 peak, same relative workload (RLT)]. Nine male subjects (178.1 ± 2.5 cm, 81.8 ± 3.3 kg, 27.4 ± 2.0 yr) were tested before and after 9 wk of cycle ergometer training, five times a week at 75%V˙o 2 peak. The power output that elicited 66.0 ± 1.1% ofV˙o 2 peak before training elicited 54.0 ± 1.7% after training. Whole body glucose Rd decreased posttraining at ABT (5.45 ± 0.31 mg ⋅ kg-1 ⋅ min-1at 65% pretraining to 4.36 ± 0.44 mg ⋅ kg-1 ⋅ min-1) but not at RLT (5.94 ± 0.47 mg ⋅ kg-1 ⋅ min-1). Net glucose uptake was attenuated posttraining at ABT (1.87 ± 0.42 mmol/min at 65% pretraining and 0.54 ± 0.33 mmol/min) but not at RLT (2.25 ± 0.81 mmol/min). The decrease in leg net glucose uptake at ABT was of similar magnitude as the drop in glucose Rd and thus could explain dampened glucose flux after training. Glycogen degradation also decreased posttraining at ABT but not RLT. Leg net glucose uptake accounted for 61% of blood glucose flux before training and 81% after training at the same relative (65%V˙o 2 peak) workload and only 38% after training at ABT. We conclude that 1) alterations in active muscle glucose uptake with training determine changes in whole body glucose kinetics; 2) muscle glucose uptake decreases for a given, moderate intensity task after training; and 3) hard exercise (65%V˙o 2 peak) promotes a glucose shunt from inactive tissues to active muscle.
Monitoring Energy Balance in Breast Cancer Survivors Using a Mobile App: Reliability Study

Background The majority of breast cancer survivors do not meet recommendations in terms of diet and physical activity. To address this problem, we developed a mobile health (mHealth) app for assessing and monitoring healthy lifestyles in breast cancer survivors, called the Energy Balance on Cancer (BENECA) mHealth system. The BENECA mHealth system is a novel and interactive mHealth app, which allows breast cancer survivors to engage themselves in their energy balance monitoring. BENECA was designed to facilitate adherence to healthy lifestyles in an easy and intuitive way. Objective The objective of the study was to assess the concurrent validity and test-retest reliability between the BENECA mHealth system and the gold standard assessment methods for diet and physical activity. Methods A reliability study was conducted with 20 breast cancer survivors. In the study, tri-axial accelerometers (ActiGraphGT3X+) were used as gold standard for 8 consecutive days, in addition to 2, 24-hour dietary recalls, 4 dietary records, and sociodemographic questionnaires. Two-way random effect intraclass correlation coefficients, a linear regression-analysis, and a Passing-Bablok regression were calculated. Results The reliability estimates were very high for all variables (alpha≥.90). The lowest reliability was found in fruit and vegetable intakes (alpha=.94). The reliability between the accelerometer and the dietary assessment instruments against the BENECA system was very high (intraclass correlation coefficient=.90). We found a mean match rate of 93.51% between instruments and a mean phantom rate of 3.35%. The Passing-Bablok regression analysis did not show considerable bias in fat percentage, portions of fruits and vegetables, or minutes of moderate to vigorous physical activity. Conclusions The BENECA mHealth app could be a new tool to measure energy balance in breast cancer survivors in a reliable and simple way. Our results support the use of this technology to not only to encourage changes in breast cancer survivors' lifestyles, but also to remotely monitor energy balance. Trial Registration ClinicalTrials.gov NCT02817724; https://clinicaltrials.gov/ct2/show/NCT02817724 (Archived by WebCite at http://www.webcitation.org/6xVY1buCc)
Lipid rather than glucose metabolism is implicated in altered insulin secretion caused by oleate in INS-1 cells.

A comprehensive metabolic study was carried out to understand how chronic exposure of pancreatic β-cells to fatty acids causes high basal secretion and impairs glucose-induced insulin release. INS-1 β-cells were exposed to 0.4 mM oleate for 3 days and subsequently incubated at 5 or 25 mM glucose, after which various parameters were measured. Chronic oleate promoted triglyceride deposition, increased fatty acid oxidation and esterification, and reduced malonyl-CoA at low glucose in association with elevated basal O2 consumption and redox state. Oleate caused a modest (25%) reduction in glucose oxidation but did not affect glucose usage, the glucose 6-phosphate and citrate contents, and the activity of pyruvate dehydrogenase of INS-1 cells. Thus changes in glucose metabolism and a Randle-glucose/fatty acid cycle do not explain the altered secretory properties of β-cells exposed to fatty acids. The main response of INS-1 cells to chronic oleate, which is to increase the oxidation and esterification of fatty acids, may contribute to cause high basal insulin secretion via increased production of reducing equivalents and/or the generation of complex lipid messenger molecule(s).
Effects of 12-week Vegetarian Diet on the Nutritional Status, Stress Status and Bowel Habits in Middle School Students and Teachers

This study investigated the effect of switching normal diet to vegetarian diet rich in vegetables and fruits for school foodservice and home meal on the nutritional status, bowel habit improvement and stress reduction of teachers and adolescents. A total of 40 research subjects (26 students, 14 teachers) from one middle school voluntarily participated in the research. Questionnaire surveys and blood analysis were conducted before and after a 12-week vegetarian diet period. The participants were asked on their dietary habit, bowel habit and stress measurement. After 12 weeks, reduction of BMI (kg/m2) in the students (p < 0.05) and reduction of TC (mg/dL) in both teachers and students (p < 0.05) were observed. Also reduction of LDL-C (mg/dL) was observed in the teachers (p < 0.05) whereas serum calcium and Vitamin B12 was increased in the students and teachers (p < 0.005). The teacher's stress level was reduced (p < 0.05) after the 12-week vegetarian diet. As for the changes in bowel habit, the number of the students and teachers classified as experiencing functional constipation was decreased respectively from 10 to 7, from 7 to 5. Based on the result, it is considered that the vegetarian diet rich in fruits and vegetables improved general health status of study subjects suggesting that such a dietary habit would substantially contribute to improving nutritional status and bowel habit.
Antimicrobial Activities of European Propolis Collected from Various Geographic Origins Alone and in Combination with Antibiotics

Background: Propolis consists of a complex mixture of resinous substances collected by honeybees from different plant sources. The objective of this study was to investigate the chemical composition, biological activities, and synergistic properties with antibiotics of propolis samples collected from various geographic origins (Germany, Ireland, and Czech Republic). Methods: The chemical composition of the propolis was analyzed by Gas Liquid Chromatography-Mass Spectrometry (GLC-MS) and High-performance liquid chromatography (HPLC). The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) were evaluated by the standard broth microdilution method, while synergistic interactions were assessed by checkerboard dilution and time-kill curve assays. Results: HPLC and GLC-MS analyses revealed that ethanol extract of propolis (EEP) and water extracts of propolis (WEP) contained more than 100 different phytochemicals. The most abundant compounds were aromatic alcohols, aromatic acids, cinnamic acid and its esters, fatty acids, and flavanone (chrysin). Czech propolis showed the highest phenolic content (129.83 ± 5.9 mg CAE/g) followed by Irish propolis and German propolis. Furthermore, Irish propolis exhibited the highest value of total flavonoid content (2.86 ± 0.2 mg QE/g) and antioxidant activity (IC50 = 26.45 µg/mL). All propolis samples showed moderate antibacterial effect against Gram-positive microorganisms with MIC ranging from 0.08 mg/mL to 2.5 mg/mL. Moreover, EEP exhibited moderate activity against Gram-negative bacteria with MIC between 0.6 mg/mL to 5 mg/mL. In addition, EEP displayed moderate antifungal activity (MIC values between 0.6–2.5 mg/mL). The results obtained from time kill-kinetic assay and checkerboard dilution test of two-drug combinations between EEP and antibiotics such as vancomycin, oxacillin, and levofloxacin indicate mainly synergistic interactions against drug-resistant microbial pathogens including MRSA and VRE. Conclusions: The propolis extract synergistically enhanced the efficacy of antibiotics, especially those acting on cell wall synthesis (vancomycin and oxacillin) against drug-resistant microorganisms.
Effect of high-altitude acclimation on NEFA turnover and lipid utilization during exercise in rats.

Relative exercise intensity (or %maximum O2 consumption,V˙o 2 max) controls fuel selection at sea level (SL) and after high-altitude acclimation (HA) in rats. In this context we used indirect calorimetry, [1-14C]palmitate infusions, and muscle triacylglycerol (TAG) measurements to determine 1) total lipid oxidation, 2) the relationship between circulatory nonesterified fatty acid (NEFA) flux and concentration, and 3) muscle TAG depletion after exercise in HA-acclimated rats. Aerobic capacity is decreased in trained rats after 10 wk of acclimation. Both SL and HA showed the same relative use of lipids at 60% [62 ± 5% (HA) and 61 ± 3% (SL) of O2 consumption (V˙o 2)] and 80% [46 ± 6% (HA) and 47 ± 5% (SL) ofV˙o 2] of their respective V˙o 2 max. At 60% V˙o 2 max, plasma [NEFA] were higher in HA, but rate of appearance was essentially the same in both groups (at 30 min, 38 ± 9 vs. 49 ± 6 μmol ⋅ kg-1 ⋅ min-1in HA and SL, respectively). At this intensity SL showed no significant decrease in muscle TAG, but in HA it decreased by 64% in soleus and by 90% in red gastrocnemius. We conclude that 1) the relative contributions of total lipid are the same in SL and HA, contrary to differences in [NEFA], because the relationship between flux rate and [NEFA] is modified after acclimation, and 2) muscle TAG may play a more important role at HA.
Ablation of the SERCA3 gene alters epithelium-dependent relaxation in mouse tracheal smooth muscle.

Sarcoplasmic/endoplasmic reticulum Ca2+-ATPase 3 (SERCA3), an isoform of the intracellular Ca2+ pump that has been shown to mediate endothelium-dependent relaxation of vascular smooth muscle, is also expressed in tracheal epithelium. To determine its possible role in regulation of airway mechanical function, we compared tracheal contractility in gene-targeted mice deficient in SERCA3 (SERCA3-) with that in wild-type tracheae. Cumulative addition of ACh elicited concentration-dependent increases in isometric force (ED50 = 2 μM, maximum force = 8 mN/mm2) that were identical in SERCA3- and wild-type tracheae. After ACh stimulation, substance P (SP) elicited a transient relaxation (42.6 ± 3.2%, n = 28) in both tracheae. However, the rate of relaxation was significantly ( P < 0.04, n = 9) more rapid in the wild-type [half-time ( t ½) = 34.3 s] than in the SERCA3-( t ½ = 61.6 s) trachea. The SP relaxation was reduced by rubbing the trachea, indicative of epithelial cell involvement. This was verified using a perfused trachea preparation. SP in the outside medium had no effect, whereas SP in the perfusate bathing the epithelial side elicited a relaxation. Nitric oxide synthase inhibition (0.2 mM N ω-nitro-l-arginine) reduced the SP relaxation by 36.5 ± 12.5%, whereas the SP effect was abolished by eicosanoid inhibition (10 μM indomethacin). ATP also elicited an epithelium-dependent relaxation similar to SP but with a more rapid relaxation in the SERCA3-trachea than in the wild-type trachea. Our results indicate that SERCA3 gene ablation does not directly affect smooth muscle, which is consistent with the distribution of the isoform, but suggest that SERCA3 plays a role in epithelial cell modulation of airway smooth muscle function.
Comprehensive formal verification of an OS microkernel

We present an in-depth coverage of the comprehensive machine-checked formal verification of seL4, a general-purpose operating system microkernel. We discuss the kernel design we used to make its verification tractable. We then describe the functional correctness proof of the kernel's C implementation and we cover further steps that transform this result into a comprehensive formal verification of the kernel: a formally verified IPC fastpath, a proof that the binary code of the kernel correctly implements the C semantics, a proof of correct access-control enforcement, a proof of information-flow noninterference, a sound worst-case execution time analysis of the binary, and an automatic initialiser for user-level systems that connects kernel-level access-control enforcement with reasoning about system behaviour. We summarise these results and show how they integrate to form a coherent overall analysis, backed by machine-checked, end-to-end theorems. The seL4 microkernel is currently not just the only general-purpose operating system kernel that is fully formally verified to this degree. It is also the only example of formal proof of this scale that is kept current as the requirements, design and implementation of the system evolve over almost a decade. We report on our experience in maintaining this evolving formally verified code base.
Interactions of keratinocyte growth factor with a nitrating species after marrow transplantation in mice.

We reported that allogeneic T cells given to irradiated mice at the time of marrow transplantation stimulated tumor necrosis factor (TNF)-α, interferon (IFN)-γ, and nitric oxide (⋅ NO) production in the lung, and the addition of cyclophosphamide (known to stimulate superoxide production) favored the generation of a nitrating species. Although keratinocyte growth factor (KGF) prevents experimental lung injury by promoting epithelial repair, its effects on the production of inflammatory mediators has not been studied. KGF given before transplantation inhibited the T cell-induced increase in bronchoalveolar lavage fluid protein, TNF-α, IFN-γ, and nitrite levels measured on day 7 after transplantation without modifying cellular infiltration or proinflammatory cytokines and inducible ⋅ NO synthase mRNA. KGF also suppressed ⋅ NO production by alveolar macrophages obtained from mice injected with T cells. In contrast, the same schedule of KGF failed to prevent permeability edema or suppress TNF-α, IFN-γ, and ⋅ NO production in mice injected with both T cells and cyclophosphamide. Because only epithelial cells respond to KGF, these data are consistent with the production of an epithelial cell-derived mediator capable of downregulating macrophage function. However, the presence of a nitrating agent impairs KGF-derived responses.
Role of Myeloid-Derived Suppressor Cells in Amelioration of Experimental Autoimmune Hepatitis Following Activation of TRPV1 Receptors by Cannabidiol

Background Myeloid-derived suppressor cells (MDSCs) are getting increased attention as one of the main regulatory cells of the immune system. They are induced at sites of inflammation and can potently suppress T cell functions. In the current study, we demonstrate how activation of TRPV1 vanilloid receptors can trigger MDSCs, which in turn, can inhibit inflammation and hepatitis. Methodology/Principal Findings Polyclonal activation of T cells, following injection of concanavalin A (ConA), in C57BL/6 mice caused acute hepatitis, characterized by significant increase in aspartate transaminase (AST), induction of inflammatory cytokines, and infiltration of mononuclear cells in the liver, leading to severe liver injury. Administration of cannabidiol (CBD), a natural non-psychoactive cannabinoid, after ConA challenge, inhibited hepatitis in a dose-dependent manner, along with all of the associated inflammation markers. Phenotypic analysis of liver infiltrating cells showed that CBD-mediated suppression of hepatitis was associated with increased induction of arginase-expressing CD11b+Gr-1+ MDSCs. Purified CBD-induced MDSCs could effectively suppress T cell proliferation in vitro in arginase-dependent manner. Furthermore, adoptive transfer of purified MDSCs into naïve mice conferred significant protection from ConA-induced hepatitis. CBD failed to induce MDSCs and suppress hepatitis in the livers of vanilloid receptor-deficient mice (TRPV1−/−) thereby suggesting that CBD primarily acted via this receptor to induce MDSCs and suppress hepatitis. While MDSCs induced by CBD in liver consisted of granulocytic and monocytic subsets at a ratio of ∼2∶1, the monocytic MDSCs were more immunosuppressive compared to granulocytic MDSCs. The ability of CBD to induce MDSCs and suppress hepatitis was also demonstrable in Staphylococcal enterotoxin B-induced liver injury. Conclusions/Significance This study demonstrates for the first time that MDSCs play a critical role in attenuating acute inflammation in the liver, and that agents such as CBD, which trigger MDSCs through activation of TRPV1 vanilloid receptors may constitute a novel therapeutic modality to treat inflammatory diseases.