Physicochemical Characterisation of Seeds, Oil and Defatted Cake of Three Hempseed Varieties Cultivated in Spain.

The increasing use of hempseed in food products highlights the need for a comprehensive database for scientific research and industrial applications. In food development, information about the techno-functional properties of raw materials plays a crucial role in determining the suitability of each product for specific applications. Thus, this study aims to characterise three hempseed varieties (Ferimon, Henola and Uso-31), comparing their physicochemical and nutritional compositions. Moreover, the study investigates the impact of hempseed varieties on the techno-functional, physical and thermal properties of the partially defatted hempseed flours (PDHFs) obtained from single screw pressing (SSP) oil extraction. The fatty acid and tocopherol profiles of the dehulled seeds and oil were also analysed. Significant variations in yield and physical properties were observed among hempseed varieties, influenced by genetics, adaptation to agro-climatic conditions and cultivation systems. Despite its lower yield (kg/ha), Uso-31 exhibited superior 1000-seed weight, dehulling yield and larger mean seed size (1.79 ± 0.02 mm). Hempseed oil was rich in unsaturated fatty acids, particularly linoleic (51.2-53.4 g/100 g oil) and α-linolenic (14.88-18.97 g/100 oil) acids, showing variations in γ- and α-tocopherols depending on the variety. The variety also influenced the least gelation concentration (LGC) and techno-functional properties such as water absorption capacity (WAC), emulsifying activity (EA) and emulsion stability (ES). SDS-PAGE and DSC measurements indicated the presence of 11S and 7S globulin proteins with denaturation temperatures above 87.8 °C. These findings confirm that the studied hempseed flours are valuable techno-functional and nutritional ingredients suitable for sustainable food formulations.

Endothelial progenitor cells as biomarkers of diabetes-related cardiovascular complications.

Diabetes mellitus (DM) constitutes a chronic metabolic disease characterized by elevated levels of blood glucose which can also lead to the so-called diabetic vascular complications (DVCs), responsible for most of the morbidity, hospitalizations and death registered in these patients. Currently, different approaches to prevent or reduce DM and its DVCs have focused on reducing blood sugar levels, cholesterol management or even changes in lifestyle habits. However, even the strictest glycaemic control strategies are not always sufficient to prevent the development of DVCs, which reflects the need to identify reliable biomarkers capable of predicting further vascular complications in diabetic patients. Endothelial progenitor cells (EPCs), widely known for their potential applications in cell therapy due to their regenerative properties, may be used as differential markers in DVCs, considering that the number and functionality of these cells are affected under the pathological environments related to DM. Besides, drugs commonly used with DM patients may influence the level or behaviour of EPCs as a pleiotropic effect that could finally be decisive in the prognosis of the disease. In the current review, we have analysed the relationship between diabetes and DVCs, focusing on the potential use of EPCs as biomarkers of diabetes progression towards the development of major vascular complications. Moreover, the effects of different drugs on the number and function of EPCs have been also addressed.

PCR-Based Strategy for Introducing CRISPR/Cas9 Machinery into Hematopoietic Cell Lines.

Acute myeloid leukemia is a complex heterogeneous disease characterized by the clonal expansion of undifferentiated myeloid precursors. Due to the difficulty in the transfection of blood cells, several hematological models have recently been developed with CRISPR/Cas9, using viral vectors. In this study, we developed an alternative strategy in order to generate CRISPR constructs by fusion PCR, which any lab equipped with basic equipment can implement. Our PCR-generated constructs were easily introduced into hard-to-transfect leukemic cells, and their function was dually validated with the addition of MYBL2 and IDH2 genes into HEK293 cells. We then successfully modified the MYBL2 gene and introduced the R172 mutation into the IDH2 gene within NB4 and HL60 cells that constitutively expressed the Cas9 nuclease. The efficiency of mutation introduction with our methodology was similar to that of ribonucleoprotein strategies, and no off-target events were detected. Overall, our strategy represents a valid and intuitive alternative for introducing desired mutations into hard-to-transfect leukemic cells without viral transduction.

Impact of Starch Concentration on the Pasting and Rheological Properties of Gluten-Free Gels. Effects of Amylose Content and Thermal and Hydration Properties.

The pasting and rheological properties of starch gels from different botanical origins have been widely used to evaluate the application of these starches in pharmaceutical and food products. However, the ways in which these properties are modified by starch concentration and their dependence on amylose content and thermal and hydration properties have not been adequately established so far. An exhaustive study of the pasting and rheological properties of starch gels (maize and rice (normal and waxy in both cases), wheat, potato, and tapioca) at concentrations of 6.4, 7.8, 9.2, 10.6, and 11.9 g/100 g was performed. The results were evaluated in terms of a potential equation fit between each parameter and each gel concentration. The parameters determined for the gels at the studied concentrations were correlated with the hydration properties and thermal properties by applying principal component analysis (PCA). Wheat starch, followed by normal maize and normal rice starches, presented a greater capacity to modulate their gels' pasting and viscoelastic properties via their concentration in water. On the contrary, the characteristics of waxy rice and maize, potato, and tapioca starches were barely modified by concentration in pasting assays, but the gels of potato and tapioca showed noticeable changes in their viscoelastic properties as functions of concentration. In the PCA plot, the non-waxy cereal samples (wheat, normal maize, and normal rice) were located close to each other. Wheat starch gels were the most dispersed on the graph, which is consistent with the high dependence on the concentration of the gel shown in most of the studied parameters. The waxy starches had close positions not too distant from those of the tapioca and potato samples and with little influence from amylose concentration. The potato and tapioca samples were close to the vectors of the crossover point in rheology and peak viscosity in their pasting properties. The knowledge gained from this work allows a better understanding of the effects of starch concentration on food formulations.

Modification of the microstructure of tef [Eragrostis tef (Zucc.) Trotter] flour ultrasonicated at different temperatures. Impact on its techno-functional and rheological properties.

Tef flour comes from a nutritionally-rich ancient grain gaining increasing interest in gluten-free market. Gluten-free sources are modified by different means to improve their functionality. Ultrasound treatment (US) alters flours' structure and leads to physically modified flours with a wider application range. The aim of the present work was to evaluate the impact of US treatments of moderate treatment time, 10 min, and high concentration of the aqueous flour dispersion, 25%, on the microstructural, starch damage, apparent amylose content, techno-functional, pasting and rheological properties of two tef flour varieties, white and brown. Temperature was varied (20, 40, 45, 50, and 55 °C) to modulate the impact of sonication. US treatments led to general particle fragmentation which markedly increased starch damage and lightness (L*) values. Apparent amylose content was higher after ultrasonication, as consequence of molecular fragmentation due to cavitation. Increased starch granules' exposed area led to enhanced interaction with water, promoting the water absorption index (WAI) and swelling power (SP) of treated flours. Pasting properties showed increased pasting temperatures as well as decreased viscometric profiles with lower breakdown viscosities, indicative of starch rearrangement improved by increasing temperature. Rheological properties indicated higher consistency in gels after US treatments, with improved ability to withstand stress and lower values of tan(δ)1 reflecting a higher solid-like behavior and higher strength of the gel. Temperature was found to be a crucial variable during US treatments, showing an improved degree of modification at higher temperatures in ultrasonicated tef flours, following the same trend in both varieties.

Impact of Solubilized Substances on the Techno-Functional, Pasting and Rheological Properties of Ultrasound-Modified Rice, Tef, Corn and Quinoa Flours.

The modification of flours by ultrasound (US) treatments requires excess water to suspend the sample to be treated, which must be removed after treatment to recover the ultrasonicated flour. The aim of this study was to determine the influence that the water removal method has on the final characteristics of US-treated gluten-free flours (rice, brown tef, corn and quinoa). US treatment parameters were constant, and two water removal methods were studied: freeze-drying and centrifugation + drying. The elimination of water by centrifugation resulted in the loss of solubilized compounds from the treated flours, which led to important differences between the final characteristics of US-treated flours. Ultrasonication resulted in the reduction of flours' particle size and modification of their color parameters. Techno-functional properties were modified by US treatment, where the water removal method was more influential in whole grain samples (brown tef and quinoa). Few differences were found in thermal properties among pairs of US-treated samples, indicative that the effect caused to starch was mainly attributed to ultrasonication conditions than to the drying method. The water removal method markedly influenced the pasting properties of US-treated flours, resulting in lower profiles when freeze-drying was applied and higher profiles when flours were retrieved by centrifugation. Gels made with tef, corn and quinoa presented reduced tan(δ)1 values after sonication, while gels made with rice did not show any modification. The water removal method is a decisive step in US treatments, defining the final characteristics of the treated matter, and having a great influence in the modification attributed to ultrasonication.

Impact of microwave radiation on in vitro starch digestibility, structural and thermal properties of rice flour. From dry to wet treatments.

Microwave radiation (MW) is an environment-friendly technology used to physically modify flours. Rice flour was MW-treated at different moisture content (MC) (3 %, 8 %, 13 %, 15 %, 20 % and 30 %). In vitro starch digestibility was determined and related to the changes caused by MW treatment to flours' structure and thermal properties, which were influenced by MC. A reduction of 49 % and 65 % in the gelatinization enthalpy of samples treated at 20 % and 30 % MC denoted a partial gelatinization. A loss of granular crystallinity in treated samples was confirmed by XR-diffraction and FTIR, particularly at 15 %, 20 % and 30 % MC. MW promoted the formation of random-coil, α-helix and ß-turn protein structure, and the disappearance of LF-ß-sheet. Morphological differences were found between samples treated at 8 % MC (loss of polygonal structure, protein layer covering granules' surface and small holes) and 30 % MC (rounded and aggregated granules, covered with exudate amylose). In vitro starch digestibility revealed that samples treated at 20 % and 30 % MC showed 40 % and 47 % higher rapidly digestible starch, 48 % and 70 % lower slowly digestible starch and 90 % lower resistant starch than the untreated flour. Flour MC in MW-treatment allowed the modulation of structural and thermal characteristics of rice flour and consequently its starch hydrolysis rate.

Assessment of endothelial colony forming cells delivery routes in a murine model of critical limb threatening ischemia using an optimized cell tracking approach.

BACKGROUND: Endothelial colony forming cells (ECFCs), alone or in combination with mesenchymal stem cells, have been selected as potential therapeutic candidates for critical limb-threatening ischemia (CLTI), mainly for those patients considered as "no-option," due to their capability to enhance revascularization and perfusion recovery of ischemic tissues. Nevertheless, prior to translating cell therapy to the clinic, biodistribution assays are required by regulatory guidelines to ensure biosafety as well as to discard undesired systemic translocations. Different approaches, from imaging technologies to qPCR-based methods, are currently applied. METHODS: In the current study, we have optimized a cell-tracking assay based on DiR fluorescent cell labeling and near-infrared detection for in vivo and ex vivo assays. Briefly, an improved protocol for DiR staining was set up, by incubation of ECFCs with 6.67 µM DiR and intensive washing steps prior cell administration. The minimal signal detected for the residual DiR, remaining after these washes, was considered as a baseline signal to estimate cell amounts correlated to the DiR intensity values registered in vivo. Besides, several assays were also performed to determine any potential effect of DiR over ECFCs functionality. Furthermore, the optimized protocol was applied in combination with qPCR amplification of specific human Alu sequences to assess the final distribution of ECFCs after intramuscular or intravenous administration to a murine model of CLTI. RESULTS: The optimized DiR labeling protocol indicated that ECFCs administered intramuscularly remained mainly within the hind limb muscle while cells injected intravenously were found in the spleen, liver and lungs. CONCLUSION: Overall, the combination of DiR labeling and qPCR analysis in biodistribution assays constitutes a highly sensitive approach to systemically track cells in vivo. Thereby, human ECFCs administered intramuscularly to CLTI mice remained locally within the ischemic tissues, while intravenously injected cells were found in several organs. Our data corroborate the need to perform biodistribution assays in order to define specific parameters such as the optimal delivery route for ECFCs before their application into the clinic.

Determination of puberty in gilts: contrast of diagnostic methods.

BACKGROUND: Early onset of a gilt´s puberty is needed for adequate economic performance in farms, because it indicates her reproductive performance and longevity. Therefore, an effective diagnosis is needed. Our purpose was to compare different procedures (external characteristics, blood progesterone analysis and ultrasonography diagnosis) to detect puberty in 70 gilts (Topigs TN70; 240 days old) on farm conditions. Postmortem examination was the standard reference. Multiple logistic regression analysis was used to identify which combination of independent variables (predictors) best predicts the status of gilts. RESULTS: Puberty (46/70 gilts; 65.71%) was characterized by the presence of follicles larger than 6 mm, corpus albicans, corpus rubrum, and corpus luteum (postmortem examination). Vaginal length, body condition, backfat, carcass weight and progesterone blood concentration were significantly higher in pubertal than prepubertal gilts (P < 0.05). Two types of ultrasonography equipment (DELTA and W3) were compared and performed by the same senior technician (V1). The results obtained by two technicians with different levels of experience (V1 and V2, a junior technician) using W3 were also compared. Ultrasonography provided better results than other diagnostic techniques, although the effectiveness of the ultrasonography changed with technological improvements and with increased expertise of technicians. The most accurate results were found by V1/DELTA (Nagelkerke´s R2 = 0.846; Sensitivity = 0.956; Specificity = 0.958; Positive predictive value = 0.978; Negative predictive value = 0.920; Area under ROC curve = 0.957). Results using the W3 equipment could be improved when used in conjunction with vaginal length (V1; Nagelkerke´s R2 = 0.834; Sensitivity = 0.933; Specificity = 0.958; Positive predictive value = 0.977; Negative predictive value = 0.885; Area under ROC curve = 0.972) or progesterone concentration (V2; Nagelkerke´s R2 = 0.780; Sensitivity = 0.955; Specificity = 0.826; Positive predictive value = 0.915; Negative predictive value = 0.905; Area under ROC curve = 0.970). CONCLUSIONS: Ultrasonography provided better results than other diagnostic techniques. The effectiveness of the ultrasonography changes with technological improvements and with increased expertise of technicians. Results using the W3 equipment could be improved when used along with vaginal length (V1) or progesterone concentration (V2). Accuracy parameters are a guide to choose puberty diagnosis, but the farms must also evaluate effect on gilts, ease and cost of administration.

Techno-Functional and Gelling Properties of Acha (Fonio) (Digitaria exilis stapf) Flour: A Study of Its Potential as a New Gluten-Free Starch Source in Industrial Applications.

Fonio (Digitaria exilis Stapf) is an ancient African cereal that represents a rich source of carbohydrate, fat, fiber, vitamins, minerals, and sulfur-containing amino acids. Processing and utilization of fonio require adequate knowledge of its structural, chemical, and nutritional characteristics. The present work evaluates the structural, techno-functional, and gelling properties of fonio and compares them to other major gluten-free cereals (rice, maize, sorghum, and millet). Fonio flour presented significantly higher water absorption index and swelling power, while it scored a lower water solubility index than the reference flours. The pasting viscosity profile of fonio was similar to that of rice, with equivalent peak viscosity but a breakdown viscosity 24% lower than rice, indicative of higher stability and resistance to shearing and heating. Rheological properties demonstrated that fonio generates gels with remarkably strong structures. At 15% concentration, fonio gel withstood stress 579% higher than those observed in the reference flours without breaking its structure. Fonio flour presented the highest gelatinization enthalpy (11.45 J/g) and a narrow gelatinization temperature range (9.96 °C), indicative of a better-packed starch structure than the other analyzed flours. The texture of the gels made with fonio showed higher firmness over the evaluated period. These combined results suggest that fonio is a suitable ingredient for gel-like food formulations.

Long Term Response to Circulating Angiogenic Cells, Unstimulated or Atherosclerotic Pre-Conditioned, in Critical Limb Ischemic Mice.

Critical limb ischemia (CLI), the most severe form of peripheral artery disease, results from the blockade of peripheral vessels, usually correlated to atherosclerosis. Currently, endovascular and surgical revascularization strategies cannot be applied to all patients due to related comorbidities, and even so, most patients require re-intervention or amputation within a year. Circulating angiogenic cells (CACs) constitute a good alternative as CLI cell therapy due to their vascular regenerative potential, although the mechanisms of action of these cells, as well as their response to pathological conditions, remain unclear. Previously, we have shown that CACs enhance angiogenesis/arteriogenesis from the first days of administration in CLI mice. Also, the incubation ex vivo of these cells with factors secreted by atherosclerotic plaques promotes their activation and mobilization. Herein, we have evaluated the long-term effect of CACs administration in CLI mice, whether pre-stimulated or not with atherosclerotic factors. Remarkably, mice receiving CACs and moreover, pre-stimulated CACs, presented the highest blood flow recovery, lower progression of ischemic symptoms, and decrease of immune cells recruitment. In addition, many proteins potentially involved, like CD44 or matrix metalloproteinase 9 (MMP9), up-regulated in response to ischemia and decreased after CACs administration, were identified by a quantitative proteomics approach. Overall, our data suggest that pre-stimulation of CACs with atherosclerotic factors might potentiate the regenerative properties of these cells in vivo.

Effects of three commercial vaccines against porcine parvovirus 1 in pregnant gilts.

Porcine parvovirosis is a common and important cause of reproductive failure in naïve dams. Even though vaccination is generally effective at preventing disease occurrence, the homology between the vaccine and challenge strains has been recently suggested to play a role in protection. Therefore, the purpose of this study was to evaluate and compare the efficacy of three currently available commercial vaccines against porcine parvovirus genotype 1 (PPV1) in an experimental model using pregnant gilts. Seventy-seven PPV1-negative gilts were included in the trial and randomly allocated to four groups. In group 1, gilts received two doses, three weeks apart, of a PPV1 subunit vaccine (ReproCyc® ParvoFLEX). Following the same scheme, gilts from group 2 received two doses of a PPV1 bivalent vaccine (ERYSENG® PARVO). In group 3, gilts received two doses, four weeks apart, of a PPV1 octavalent vaccine (Porcilis® Ery + Parvo + Lepto). Lastly, gilts from group 4 were left untreated and were used as challenge controls. All gilts were artificially inseminated three weeks after completion of vaccination. Pregnant animals were subsequently challenged around 40 days of gestation with a heterologous PPV1 strain. Foetuses were harvested at around day 90 of gestation and evaluated for their macroscopic appearance (i.e., normal, mummified, or autolytic). Along the study, safety parameters after vaccination, antibody responses against PPV1 and viremia in gilts were also measured. All the foetuses in the challenge control group were mummified, which validated the challenge model, whereas the three evaluated vaccines protected the progeny against PPV1 by preventing the appearance of clinical manifestations associated to parvovirosis. Remarkably, the PPV1 subunit vaccine induced an earlier seroconversion of gilts and was the only vaccine that could prevent viremia after challenge. This vaccine also achieved the largest average litter size accompanied with a high average proportion of clinically healthy foetuses.

REX-001, a BM-MNC Enriched Solution, Induces Revascularization of Ischemic Tissues in a Murine Model of Chronic Limb-Threatening Ischemia.

Background: Bone Marrow Mononuclear Cells (BM-MNC) constitute a promising alternative for the treatment of Chronic Limb-Threatening ischemia (CLTI), a disease characterized by extensive blockade of peripheral arteries, clinically presenting as excruciating pain at rest and ischemic ulcers which may lead to gangrene and amputation. BM-MNC implantation has shown to be efficient in promoting angiogenesis and ameliorating ischemic symptoms in CLTI patients. However, the variability seen between clinical trials makes necessary a further understanding of the mechanisms of action of BM-MNC, and moreover, to improve trial characteristics such as endpoints, inclusion/exclusion criteria or drug product compositions, in order to implement their use as stem-cell therapy. Materials: Herein, the effect of REX-001, a human-BM derived cell suspension enriched for mononuclear cells, granulocytes and CD34+ cells, has been assessed in a murine model of CLTI. In addition, a REX-001 placebo solution containing BM-derived red blood cells (BM-RBCs) was also tested. Thus, 24 h after double ligation of the femoral artery, REX-001 and placebo were administrated intramuscularly to Balb-c nude mice (n:51) and follow-up of ischemic symptoms (blood flow perfusion, motility, ulceration and necrosis) was carried out for 21 days. The number of vessels and vascular diameter sizes were measured within the ischemic tissues to evaluate neovascularization and arteriogenesis. Finally, several cell-tracking assays were performed to evaluate potential biodistribution of these cells. Results: REX-001 induced a significant recovery of blood flow by increasing vascular density within the ischemic limbs, with no cell translocation to other organs. Moreover, cell tracking assays confirmed a decrease in the number of infused cells after 2 weeks post-injection despite on-going revascularization, suggesting a paracrine mechanism of action. Conclusion: Overall, our data supported the role of REX-001 product to improve revascularization and ischemic reperfusion in CLTI.

Atherosclerotic Pre-Conditioning Affects the Paracrine Role of Circulating Angiogenic Cells Ex-Vivo.

In atherosclerosis, circulating angiogenic cells (CAC), also known as early endothelial progenitor cells (eEPC), are thought to participate mainly in a paracrine fashion by promoting the recruitment of other cell populations such as late EPC, or endothelial colony-forming cells (ECFC), to the injured areas. There, ECFC replace the damaged endothelium, promoting neovascularization. However, despite their regenerative role, the number and function of EPC are severely affected under pathological conditions, being essential to further understand how these cells react to such environments in order to implement their use in regenerative cell therapies. Herein, we evaluated the effect of direct incubation ex vivo of healthy CAC with the secretome of atherosclerotic arteries. By using a quantitative proteomics approach, 194 altered proteins were identified in the secretome of pre-conditioned CAC, many of them related to inhibition of angiogenesis (e.g., endostatin, thrombospondin-1, fibulins) and cell migration. Functional assays corroborated that healthy CAC released factors enhanced ECFC angiogenesis, but, after atherosclerotic pre-conditioning, the secretome of pre-stimulated CAC negatively affected ECFC migration, as well as their ability to form tubules on a basement membrane matrix assay. Overall, we have shown here, for the first time, the effect of atherosclerotic factors over the paracrine role of CAC ex vivo. The increased release of angiogenic inhibitors by CAC in response to atherosclerotic factors induced an angiogenic switch, by blocking ECFC ability to form tubules in response to pre-conditioned CAC. Thus, we confirmed here that the angiogenic role of CAC is highly affected by the atherosclerotic environment.

Identification of the initial molecular changes in response to circulating angiogenic cells-mediated therapy in critical limb ischemia.

BACKGROUND: Critical limb ischemia (CLI) constitutes the most aggressive form of peripheral arterial occlusive disease, characterized by the blockade of arteries supplying blood to the lower extremities, significantly diminishing oxygen and nutrient supply. CLI patients usually undergo amputation of fingers, feet, or extremities, with a high risk of mortality due to associated comorbidities. Circulating angiogenic cells (CACs), also known as early endothelial progenitor cells, constitute promising candidates for cell therapy in CLI due to their assigned vascular regenerative properties. Preclinical and clinical assays with CACs have shown promising results. A better understanding of how these cells participate in vascular regeneration would significantly help to potentiate their role in revascularization. Herein, we analyzed the initial molecular mechanisms triggered by human CACs after being administered to a murine model of CLI, in order to understand how these cells promote angiogenesis within the ischemic tissues. METHODS: Balb-c nude mice (n:24) were distributed in four different groups: healthy controls (C, n:4), shams (SH, n:4), and ischemic mice (after femoral ligation) that received either 50 µl physiological serum (SC, n:8) or 5 × 105 human CACs (SE, n:8). Ischemic mice were sacrificed on days 2 and 4 (n:4/group/day), and immunohistochemistry assays and qPCR amplification of Alu-human-specific sequences were carried out for cell detection and vascular density measurements. Additionally, a label-free MS-based quantitative approach was performed to identify protein changes related. RESULTS: Administration of CACs induced in the ischemic tissues an increase in the number of blood vessels as well as the diameter size compared to ischemic, non-treated mice, although the number of CACs decreased within time. The initial protein changes taking place in response to ischemia and more importantly, right after administration of CACs to CLI mice, are shown. CONCLUSIONS: Our results indicate that CACs migrate to the injured area; moreover, they trigger protein changes correlated with cell migration, cell death, angiogenesis, and arteriogenesis in the host. These changes indicate that CACs promote from the beginning an increase in the number of vessels as well as the development of an appropriate vascular network.

Burnout Syndrome and Increased Insulin Resistance.

OBJECTIVE: Our aim was to measure the association of burnout syndrome with insulin resistance in the context of a workplace health intervention. METHODS: One-year intervention program (2015 to 2016) within a university workplace. Participants (n=55) were categorized by the presence or absence of burnout syndrome at baseline using the Maslach Burnout Inventory. Insulin resistance was calculated by the triglyceride glucose index (TyG). The Mediterranean Diet adherence score and several fitness tests were completed by the participants. RESULTS: Although participants with prevalent burnout syndrome at baseline improved their physical fitness and diet scores more than participants without burnout syndrome, multiple linear regression analyses showed that participants with prevalent burnout syndrome at baseline had increased TyG index compared with participants without burnout syndrome (ß=0.18; 95% CI, 0.01 to 0.34). CONCLUSION: Burnout syndrome may be associated with insulin resistance, despite improvements in diet and fitness.

IFNγ induces epigenetic programming of human T-bethi B cells and promotes TLR7/8 and IL-21 induced differentiation.

Although B cells expressing the IFNγR or the IFNγ-inducible transcription factor T-bet promote autoimmunity in Systemic Lupus Erythematosus (SLE)-prone mouse models, the role for IFNγ signaling in human antibody responses is unknown. We show that elevated levels of IFNγ in SLE patients correlate with expansion of the T-bet expressing IgDnegCD27negCD11c+CXCR5neg (DN2) pre-antibody secreting cell (pre-ASC) subset. We demonstrate that naïve B cells form T-bethi pre-ASCs following stimulation with either Th1 cells or with IFNγ, IL-2, anti-Ig and TLR7/8 ligand and that IL-21 dependent ASC formation is significantly enhanced by IFNγ or IFNγ-producing T cells. IFNγ promotes ASC development by synergizing with IL-2 and TLR7/8 ligands to induce genome-wide epigenetic reprogramming of B cells, which results in increased chromatin accessibility surrounding IRF4 and BLIMP1 binding motifs and epigenetic remodeling of IL21R and PRDM1 loci. Finally, we show that IFNγ signals poise B cells to differentiate by increasing their responsiveness to IL-21.

Topical Quercetin and Resveratrol Protect the Ocular Surface in Experimental Dry Eye Disease.

Purpose: To determine the anti-inflammatory effect of quercetin (QCT), resveratrol (RES), and their combination in a dry eye disease (DED) model. Methods: 0.01% QCT, 0.1% RES, 0.01% QCT + 0.1% RES (QCT + RES) or vehicle were topically applied in a desiccating stress (DS) mice model. CD4+ T cells isolated from DS-exposed mice were transferred to athymic recipient mice. Corneal fluorescein staining, tear production, and tear cytokine levels were evaluated in DS-exposed mice, and conjunctival CD4+ T cell infiltration was evaluated in recipient mice. Results: QCT (p < 0.001) and QCT + RES (p < 0.05) reduced corneal staining in DS-exposed mice. IL-1α tear concentration was reduced by QCT, RES, and QCT + RES (p < 0.05, 0.01 and 0.01, respectively) compared to DS + vehicle mice. CD4+ T cells increased in recipients of DS-exposed mice (p < 0.05) and were lower in recipients of QCT- and RES-treated mice (p < 0.05). Conclusion: The anti-inflammatory effect of QCT, RES, and QCT + RES on DED-experimental model suggests that their topical application could be used for DED treatment.

CD38 promotes pristane-induced chronic inflammation and increases susceptibility to experimental lupus by an apoptosis-driven and TRPM2-dependent mechanism.

In this study, we investigated the role of CD38 in a pristane-induced murine model of lupus. CD38-deficient (Cd38-/-) but not ART2-deficient (Art2-/-) mice developed less severe lupus compared to wild type (WT) mice, and their protective phenotype consisted of (i) decreased IFN-I-stimulated gene expression, (ii) decreased numbers of peritoneal CCR2hiLy6Chi inflammatory monocytes, TNF-α-producing Ly6G+ neutrophils and Ly6Clo monocytes/macrophages, (iii) decreased production of anti-single-stranded DNA and anti-nRNP autoantibodies, and (iv) ameliorated glomerulonephritis. Cd38-/- pristane-elicited peritoneal exudate cells had defective CCL2 and TNF-α secretion following TLR7 stimulation. However, Tnf-α and Cxcl12 gene expression in Cd38-/- bone marrow (BM) cells was intact, suggesting a CD38-independent TLR7/TNF-α/CXCL12 axis in the BM. Chemotactic responses of Cd38-/- Ly6Chi monocytes and Ly6G+ neutrophils were not impaired. However, Cd38-/- Ly6Chi monocytes and Ly6Clo monocytes/macrophages had defective apoptosis-mediated cell death. Importantly, mice lacking the cation channel TRPM2 (Trpm2-/-) exhibited very similar protection, with decreased numbers of PECs, and apoptotic Ly6Chi monocytes and Ly6Clo monocytes/macrophages compared to WT mice. These findings reveal a new role for CD38 in promoting aberrant inflammation and lupus-like autoimmunity via an apoptosis-driven mechanism. Furthermore, given the implications of CD38 in the activation of TRPM2, our data suggest that CD38 modulation of pristane-induced apoptosis is TRPM2-dependent.

A comparative study of the visual outcomes between a new trifocal and an extended depth of focus intraocular lens.

PURPOSE: To evaluate and compare the visual outcomes and ocular optical performance of the PanOptix trifocal intraocular lens (IOL) and Symfony extended range of vision IOL. METHODS: Sixty-eight eyes of 34 patients were divided into 2 groups: 20 patients with the PanOptix IOL and 14 patients with the Symfony IOL. Binocular uncorrected distance visual acuity, best-corrected distance visual acuity (BCDVA), distance-corrected intermediate visual acuity (DCIVA) at 80 and 60 cm, and distance-corrected near visual acuity (DCNVA) at 40 cm were evaluated. Additionally, preferred reading distance with best-corrected distance and visual acuity at that distance, binocular defocus curves, mesopic and photopic contrast sensitivity, photic phenomena, and monocular total higher order aberrations (HOAs) were also measured. RESULTS: The visual outcomes for PanOptix and Symfony IOL groups, respectively, were as follows: BCDVA: -0.03 ± 0.03 and -0.02 ± 0.03 logMAR; DCIVA at 80 cm: 0.06 ± 0.06 and 0.06 ± 0.04 logMAR; DCIVA at 60 cm: 0.06 ± 0.10 and 0.05 ± 0.04 logMAR; DCNVA: 0.04 ± 0.06 and 0.20 ± 0.07 logMAR (p<0.001). Similar preferred reading distances were found for both groups (37.0 ± 4.6 and 38.9 ± 5.7 cm, respectively). The visual acuities at those distances were 0.09 ± 0.08 and 0.19 ± 0.08 logMAR (p<0.001), respectively. The defocus curves showed significantly better outcomes for the PanOptix IOL from -2.0 to -4.0 D (p<0.001). No significant differences were found for contrast sensitivity, halometry, or HOAs between the groups. CONCLUSIONS: The PanOptix and Symfony IOLs showed comparable visual performance at distance and intermediate. However, the PanOptix IOL provided better near and preferred reading distance VAs and showed a more continuous range of vision than the Symfony IOL.