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The spleen is a hematopoietic organ that participates in cellular and humoral immunity. It also serves as a quality control mechanism for removing senescent and/or poorly deformable red blood cells (RBCs) from circulation. Pitting is a specialized process by which the spleen extracts particles, including malaria parasites, from within circulating RBCs during their passage through the interendothelial slits (IES) in the splenic cords. To study this physiological function in vitro, we have developed two microfluidic devices modeling the IES, according to the hypothesis that at a certain range of mechanical stress on the RBC, regulated through both slit size and blood flow, would force it undergo the pitting process without affecting the cell integrity. To prove its functionality in replicating pitting of malaria parasites, we have performed a characterization of P. falciparum-infected RBCs (P.f.-RBCs) after their passage through the devices, determining hemolysis and the proportion of once-infected RBCs (O-iRBCs), defined by the presence of a parasite antigen and absence of DAPI staining of parasite DNA using a flow cytometry-based approach. The passage of P.f.-RBCs through the devices at the physiological flow rate did not affect cell integrity and resulted in an increase of the frequency of O-iRBCs. Both microfluidic device models were capable to replicate the pitting of P.f.-RBCs ex vivo by means of mechanical constraints without cellular involvement, shedding new insights on the role of the spleen in the pathophysiology of malaria., (© 2021. The Author(s).)

The improvement of cancer therapy efficacy, the extension of patient survival and the reduction of adverse side effects are major challenges in cancer research. Targeting blood vessels has been considered a promising strategy in cancer therapy. Since the tumor vasculature is disorganized, leaky and triggers immunosuppression and tumor hypoxia, several strategies have been studied to modify tumor vasculature for cancer therapy improvement. Anti-angiogenesis was first described as a mechanism to prevent the formation of new blood vessels and prevent the oxygen supply to tumor cells, showing numerous limitations. Vascular normalization using low doses of anti-angiogenic drugs was purposed to overcome the limitations of anti-angiogenic therapies. Other strategies such as vascular promotion or the induction of high endothelial venules are being studied now to improve cancer therapy. Bone morphogenetic protein 9 (BMP9) exerts a dual effect through the activin receptor-like kinase 1 (ALK1) receptor in blood vessel maturation or activation phase of angiogenesis. Thus, it is an interesting pathway to target in combination with chemotherapies or immunotherapies. This review manuscript explores the effect of the BMP9-ALK1 pathway in tumor angiogenesis and the possible usefulness of targeting this pathway in anti-angiogenesis, vascular normalization or vascular promotion therapies.

The purpose of this work is to develop a hematocrit-independent method for the detection of beta-thalassemia trait (β-TT) and iron deficiency anemia (IDA), through the rheological characterization of whole blood samples from different donors. The results obtained herein are the basis for the development of a front microrheometry point-of-care device for the diagnosis and clinical follow-up of β-TT patients suffering hematological diseases and alterations in the morphology of the red blood cell (RBC). The viscosity is calculated as a function of the mean front velocity by detecting the sample fluid-air interface advancing through a microfluidic channel. Different viscosity curves are obtained for healthy donors, β-TT and IDA samples. A mathematical model is introduced to compare samples of distinct hematocrit, classifying the viscosity curve patterns with respect to the health condition of blood. The viscosity of the fluid at certain shear rate values varies depending on several RBC factors such as shape and size, hemoglobin (Hb) content, membrane rigidity and hematocrit concentration. Blood and plasma from healthy donors are used as reference. To validate their potential clinical value as a diagnostic tool, the viscosity results are compared to those obtained by the gold-standard method for RBC deformability evaluation, the Laser-Optical Rotational Red Cell Analyzer (LoRRCA)., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The handling editor declared a past collaboration with one of the author, J-LV-C., (Copyright © 2021 Méndez-Mora, Cabello-Fusarés, Ferré-Torres, Riera-Llobet, Krishnevskaya, Trejo-Soto, Payán-Pernía, Hernández-Rodríguez, Morales-Indiano, Alarcón, Vives-Corrons and Hernandez-Machado.)

Oligochaetes feed on bulk sediment and penetrate the sediment through the construction of burrows, making them especially vulnerable to sediment metal contamination. However, the few oligochaete species that have been tested to date are almost exclusively temperate test species. Although the warmwater adapted species Branchiura sowerbyi has been indicated as a promising candidate for tropical sediment toxicity testing, few (especially chronic) studies have been conducted so far to confirm this. Therefore, the aim of the present study was to evaluate the bioaccumulation and chronic 28d lethal and sublethal toxicity of arsenic (As) and zinc (Zn) to both the warmwater-adapted B. sowerbyi and the coldwater-adapted oligochaete Tubifex tubifex for comparison. Arsenic was more toxic to both oligochaete species than Zn. Inter- and intra-species variability in toxicity values of the two test species and other benthic invertebrates was within an order of magnitude. However, B. sowerbyi was the most sensitive species to As even for sediment concentration (EC 50 : 36.6 ± 2.1 µg/g and 147.1 ± 21.7 µg/g, for B. sowerbyi and T. tubifex, respectively) and for tissue concentration (ER 50 : 9.2 ± 0.9 µg/g and 887.0 ± 35.0 µg/g, for B. sowerbyi and T. tubifex, respectively). Finally, the Tissue Residue-effects Approach (TRA) using Effective Tissue Residues appears to be a promising way forward in advancing in this since it considers internal body concentrations., (Copyright © 2021. Published by Elsevier B.V.)

Objective: Tofacitinib (TOF) is the first Janus kinase (JAK) inhibitor approved for psoriatic arthritis (PsA). It has shown efficacy in patients refractory to anti-tumor necrosis factor-α in randomized controlled trials (RCTs). Our aim was to assess efficacy and safety of TOF in clinical practice., Methods: This was an observational, open-label multicenter study of PsA patients treated with TOF due to inefficacy or adverse events of previous therapies. Outcome variables were efficacy, corticosteroid dose-sparing effect, retention rate, and safety. A comparative study of clinical features between our cohort of patients and those from the OPAL Beyond trial was performed., Results: There were 87 patients (28 women/59 men), with a mean age of 52.8 ± 11.4 years. All patients were refractory to biologic disease-modifying antirheumatic drugs (DMARDs) and/or to conventional synthetic DMARDs plus apremilast. TOF was started at 5 mg twice daily after a mean follow-up of 12.3 ± 9.3 years from PsA diagnosis. At first month, Disease Activity Score in 28 joints based on erythrocyte sedimentation rate (DAS28-ESR) decreased from median 4.8 (IQR 4.1-5.4) to 3.7 (IQR 2.8-4.7, P < 0.01), Disease Activity Index for Psoriatic Arthritis from median 28 (IQR 18.4-34.1) to 15.5 (IQR 10.1-25.7, P < 0.01), and C-reactive protein from median 1.9 (IQR 0.3-5.0) to 0.5 (IQR 0.1-2.2) mg/dL ( P < 0.01). Also, TOF led to a significant reduction in prednisone dose. Mild adverse effects were reported in 21 patients (24.13%), mainly gastrointestinal symptoms. TOF retention rate at Month 6 was 77% (95% CI 65.2-86.3). Patients in clinical practice were older with longer disease duration and received biologic agents more commonly than those in the OPAL Beyond trial., Conclusion: Data from clinical practice confirm that TOF seems to be effective, rapid, and relatively safe in refractory PsA despite clinical differences with patients in RCTs., (Copyright © 2021 by the Journal of Rheumatology.)

Solar grade silicon (SoGSi) is a key material for the development of crystalline silicon photovoltaics (PV), which is expected to reach the tera-watt level in the next years and around 50TW in 2050. Upgraded metallurgical grade silicon (UMGSi) has already demonstrated to be a viable alternative to standard polysilicon in terms of cost and quality. This study presents the life cycle assessment (LCA) of UMG obtained by the FerroSolar process. Moreover, it shows the environmental impacts of PV modules and electricity generation based on this material. For this, an exhaustive review of the life cycle inventory (LCI) of PV value chain, from metallurgical grade silicon (MG-Si) down to electricity generation, has been carried out updating inputs for all processes. The Balance of System (BoS) has also been updated with current state of the art data for a fixed open ground large PV site (100 MW pk ). Two different electricity mixes, with low and high carbon intensities, have been considered. The results reveal that for PV electricity generation using UMG-Si instead of polysilicon leads to an overall reduction of Climate change (CC) emissions of over 20%, along with an improvement of the Energy Payback Time (EPBT) of 25%, achieving significantly low values, 12 gCO 2eq /kWh e and 0.52 years, respectively. Moreover, it is shown that UMG silicon feedstock is not the main contributor to the carbon and energy footprint of the produced electricity, leaving the first place to PV module manufacturing., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Omega-3 polyunsaturated fatty acids are associated with a lower risk of cardiometabolic diseases. However, docosahexaenoic acid (DHA) is easily oxidized, leading to cellular damage. The present study examined the effects of an increased concentration of DHA in fish oil (80% of total fatty acids) on cardiometabolic risk factors and oxidative stress compared to coconut oil, soybean oil, and fish oil containing eicosapentaenoic acid (EPA) and DHA in a balanced ratio. Forty healthy male Sprague-Dawley rats were supplemented with corresponding oil for 10 weeks. Supplementation with the fish oil containing 80% DHA decreased plasma fat, plasma total cholesterol and muscle fat compared to the coconut oil and the soybean oil. Increasing concentrations of DHA induced incorporation of DHA and EPA in cell membranes and tissues along with a decrease in ω-6 arachidonic acid. The increase in DHA promoted lipid peroxidation, protein carbonylation and antioxidant response. Taken together, the increased concentration of DHA in fish oil reduced fat accumulation compared to the coconut oil and the soybean oil. This benefit was accompanied by high lipid peroxidation and subsequent protein carbonylation in plasma and in liver. In our healthy framework, the slightly higher carbonylation found after receiving fish oil containing 80% DHA might be a protecting mechanism, which fit with the general improvement of antioxidant defense observed in those rats.

Background: The effect of renin-angiotensin system (RAS) blockade either by angiotensin-converting enzyme inhibitors (ACEis) or angiotensin-receptor blockers (ARBs) on coronavirus disease 2019 (COVID-19) susceptibility, mortality and severity is inadequately described. We examined the association between RAS blockade and COVID-19 diagnosis and prognosis in a large population-based cohort of patients with hypertension (HTN)., Methods: This is a cohort study using regional health records. We identified all individuals aged 18-95 years from 87 healthcare reference areas of the main health provider in Catalonia (Spain), with a history of HTN from primary care records. Data were linked to COVID-19 test results, hospital, pharmacy and mortality records from 1 March 2020 to 14 August 2020. We defined exposure to RAS blockers as the dispensation of ACEi/ARBs during the 3 months before COVID-19 diagnosis or 1 March 2020. Primary outcomes were: COVID-19 infection and severe progression in hospitalized patients with COVID-19 (the composite of need for invasive respiratory support or death). For both outcomes and for each exposure of interest (RAS blockade, ACEi or ARB) we estimated associations in age-, sex-, healthcare area- and propensity score-matched samples., Results: From a cohort of 1 365 215 inhabitants we identified 305 972 patients with HTN history. Recent use of ACEi/ARBs in patients with HTN was associated with a lower 6-month cumulative incidence of COVID-19 diagnosis {3.78% [95% confidence interval (CI) 3.69-3.86%] versus 4.53% (95% CI 4.40-4.65%); P < 0.001}. In the 12 344 patients with COVID-19 infection, the use of ACEi/ARBs was not associated with a higher risk of hospitalization with need for invasive respiratory support or death [OR = 0.91 (0.71-1.15); P = 0.426]., Conclusions: RAS blockade in patients with HTN is not associated with higher risk of COVID-19 infection or with a worse progression of the disease., (© The Author(s) 2021. Published by Oxford University Press on behalf of ERA.)

Chronic myeloid leukemia (CML) is a hematopoietic malignancy produced by a unique oncogenic event involving the constitutively active tyrosine-kinase (TK) BCR/ABL1 . TK inhibitors (TKI) changed its prognosis and natural history. Unfortunately, ABL1 remains unaffected by TKIs. Leukemic stem cells (LSCs) remain, and resistant mutations arise during treatment. To address this problem, we have designed a therapeutic CRISPR-Cas9 deletion system targeting BCR/ABL1 . The system was efficiently electroporated to cell lines, LSCs from a CML murine model, and LSCs from CML patients at diagnosis, generating a specific ABL1 null mutation at high efficiency and allowing the edited leukemic cells to be detected and tracked. The CRISPR-Cas9 deletion system triggered cell proliferation arrest and apoptosis in murine and human CML cell lines. Patient and murine-derived xenografts with CRISPR-edited LSCs in NOD SCID gamma niches revealed that normal multipotency and repopulation ability of CRISPR edited LSCs were fully restored. Normal hematopoiesis was restored, avoiding myeloid bias. To the best of our knowledge, we show for the first time how a CRISPR-Cas9 deletion system efficiently interrupts BCR/ABL1 oncogene in primary LSCs to bestow a therapeutic benefit. This study is a proof of concept for genome editing in all those diseases, like CML, sustained by a single oncogenic event, opening up new therapeutic opportunities.

Human cell survival requires function of the Na + /K + pump; the heteromeric protein that hydrolyzes ATP to extrude Na + and import K + across the plasmalemma, thereby building and maintaining these ions' electrochemical gradients. Numerous dominant diseases caused by mutations in genes encoding for Na + /K + pump catalytic (α) subunit isoforms highlight the importance of this protein. Here, we review literature describing disorders caused by missense mutations in ATP1A1 , the gene encoding the ubiquitously expressed α1 isoform of the Na + /K + pump. These various maladies include primary aldosteronism with secondary hypertension, an endocrine syndrome, Charcot-Marie-Tooth disease, a peripheral neuropathy, complex spastic paraplegia, another neuromuscular disorder, as well as hypomagnesemia accompanied by seizures and cognitive delay, a condition affecting the renal and central nervous systems. This article focuses on observed commonalities among these mutations' functional effects, as well as on the special characteristics that enable each particular mutation to exclusively affect a certain system, without affecting others. In this respect, it is clear how somatic mutations localized to adrenal adenomas increase aldosterone production without compromising other systems. However, it remains largely unknown how and why some but not all de novo germline or familial mutations (where the mutant must be expressed in numerous tissues) produce a specific disease and not the other diseases. We propose hypotheses to explain this observation and the approaches that we think will drive future research on these debilitating disorders to develop novel patient-specific treatments by combining the use of heterologous protein-expression systems, patient-derived pluripotent cells, and gene-edited cell and mouse models.

Development of sound criteria for metal and metalloid bioaccumulation risk assessment in river basins affected by mining activities is a necessary tool to protect the aquatic communities. The aim of this study is to propose integrative scores for tissue residues that are suitable for surveillance programs and readily interpreted in terms of risk assessment in mining impacted rivers. Tissue residues of 7 trace metals and 2 metalloids were measured in ten macroinvertebrate taxa from the Nalón River basin (Spain), affected by Hg, Cu and Au mining activities. Compared with reference sites, biomonitor taxa from Hg and Au mining districts showed the highest bioaccumulation. However, low or non-significant bioaccumulation was found in sites influenced by historical Cu mining. Multivariate analyses (ANOSIM) performed on individual taxa revealed significant differences in tissue residues between sites classified according to their ecological status. The bioaccumulation risk assessment was based on the average ratio of the actual metal tissue residues in each macroinvertebrate taxon to the corresponding Ecological Threshold tissue concentration (Tissue residue Ratio to Threshold, TRT). The suitability of the biomonitors was evaluated using linear regression models fitted to the relationships between TRT scores and site sediment pollution or ecological status scores. Biomonitor selection also considered differences in invertebrate functional traits, which can influence metal and metalloid bioavailability. Site bioaccumulation risk was assessed on an Integrated Tissue concentration score (INTISS), calculated over a selection of the most relevant chemicals (As, Cu and Hg) and 3 biomonitor taxa (Baetidae, Hydropsychidae, Microdrile oligochaetes) comprising a set of feeding styles. Based on INTISS, it was possible to predict community alteration scores, using linear regression models. A comparison of site bioaccumulation and ecological status assessments based on the departure from reference conditions showed that operational monitoring programs in basins impaired by mining can be optimized by combining both approaches., (Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.)

Rhipicephalus microplus (Canestrini, 1888) is one of the species with medical and economic relevance that has been reported in the list of Cuban tick species. Some morphological characterizations about the R. microplus species in Cuba have been published; however, molecular studies are lacking. Molecular phylogenetic analyses have grouped R. annulatus, R. australis and three clades of R. microplus in a complex named R. microplus. The present study aimed to characterize two R. microplus tick isolates, established as colonies at the Cuban National Laboratory of Parasitology. Morphological characterization of adult specimens was carried out by using Scanning Electron Microscopy. The sequences of mitochondrial genes: 12S rRNA, 16S rRNA and the subunit I of cytochrome c oxidase (coxI) and one nuclear sequence: internal transcribed spacer 2 (its2) were used for phylogenetic analyses. The life cycle under laboratory conditions for both isolates was also characterized. Tick specimens of both colonies showed morphological characteristics comparable with those distinctive for the R. microplus species. Phylogenies based on mitochondrial gene sequences identified congruently the Cuban tick colonies within the clade A of R. microplus. The life cycle of both isolates under laboratory conditions lasted 65 ± 5 days and the reproductive performance of female ticks of each colony also were similar with approximately 2500 larvae obtained from fully engorged female ticks. This study constitutes the first molecular characterization of ticks from the R. microplus species in Cuba., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The motivation for this study was to develop a microdevice for the precise rheological characterization of biofluids, especially blood. The method presented was based on the principles of rheometry and fluid mechanics at the microscale. Traditional rheometers require a considerable amount of space, are expensive, and require a large volume of sample. A mathematical model was developed that, combined with a proper experimental model, allowed us to characterize the viscosity of Newtonian and non-Newtonian fluids at different shear rates. The technology presented here is the basis of a point-of-care device capable of describing the nonlinear rheology of biofluids by the fluid/air interface front velocity characterization through a microchannel. The proposed microrheometer uses a small amount of sample to deliver fast and accurate results, without needing a large laboratory space. Blood samples from healthy donors at distinct hematocrit percentages were the non-Newtonian fluid selected for the study. Water and plasma were employed as testing Newtonian fluids for validation of the system. The viscosity results obtained for the Newtonian and non-Newtonian fluids were consistent with pertinent studies cited in this paper. In addition, the results achieved using the proposed method allowed distinguishing between blood samples with different characteristics.

Background: Vitamin D (VD) deficiency has been inversely associated with parathyroid hormone (PTH) levels in the adult population but not in children and adolescents. This study aimed to report VD concentration and its correlation with PTH levels in a sample of healthy Mexican children., Methods: We conducted a cross-sectional study with 275 healthy Mexican subjects aged 2 to 17 years to estimate the status of 25-(OH)-D and its correlation with PTH levels. The 25-(OH)-D levels were estimated by liquid chromatography-tandem mass spectrometry and PTH by radioimmunoassay., Results: Subjects were categorized as young children (2 to 5 years), school children (6 to 10 years), and adolescents (11 to 17 years). The median concentration of 25-(OH)-D in young children was 27.4 ng/ml; in school children, 25.6 ng/ml; and adolescents, 24.7 ng/ml. VD levels < 20 ng/ml were found in only 10.5% of the participants. Only 3% of the young children showed VD deficiency, in contrast to 10% of school children and 21% of adolescents (p ≤ 0.05). PTH was found within normal ranges in 95.6% ofthe studied population. VD levels < 20 mg/dl were found in 25.8% of children with overweight or obesity (p = 0.009)., Conclusions: VD levels < 20 ng/ml were observed in 10% of the studied group, but this percentage increased with age:21% of the adolescents showed VD levels < 20 ng/ml. No correlation with PTH levels was found. The VD values reported inthis study are lower than those previously reported in Mexican children.

Purpose: The optimal mode of long-term positive airway pressure (PAP) treatment for obesity-hypoventilation syndrome (OHS) is not clear. The objectives of this study were to analyze whether or not patients with severe OHS treated with continuous positive airway pressure (CPAP) remained controlled with this therapy over the long term and to investigate which variables were associated with CPAP failure and the need to switch to non-invasive ventilation (NIV)., Methods: In a retrospective single-center study, patients admitted to the hospital because of severe OHS between 1996 and 2015 were analyzed. A multiple regression analysis was performed in order to determine which variables were associated with either CPAP success or failure to maintain long-term control., Results: Of 126 consecutive patients, 115 accepted long-term PAP treatment. CPAP or NIV treatment was prescribed according to a protocol that included overnight polysomnographic PAP titration. Follow-up time was 8.0 ± 4.8 years. At the end of this period, 29% of CPAP-treated patients had been re-assigned to NIV because of recurrence of global respiratory failure. High levels of obesity, weight gain, lower FEV1/FVC values and the need for nocturnal supplementary oxygen independently predicted CPAP failure., Conclusion: CPAP therapy for severe OHS in patients who have these risk factors should be closely monitored in the long-term for possible treatment failure.

Objective: Cancer care has been disrupted by the response of health systems to the COVID-19 pandemic, especially during lockdowns. The objective of our study is to evaluate the impact of the pandemic on the incidence of cancer diagnoses in primary care., Design: Time-series study of malignant neoplasms and diagnostic procedures, using data from the primary care electronic health records from January 2014 to September 2020., Setting: Primary care, Catalonia, Spain., Participants: People older than 14 years and assigned in one of the primary care practices of the Catalan Institute of Health with a new diagnosis of malignant neoplasm., Main Outcome Measures: We obtained the monthly expected incidence of malignant neoplasms using a temporary regression, where the response variable was the incidence of cancer from 2014 to 2018 and the adjustment variables were the trend and seasonality of the time series. Excess or lack of malignant neoplasms was defined as the number of observed minus expected cases, globally and stratified by sex, age, type of cancer and socioeconomic status., Results: Between March and September 2020 we observed 8766 (95% CI 4135 to 13 397) fewer malignant neoplasm diagnoses, representing a reduction of 34% (95% CI 19.5% to 44.1%) compared with the expected. This underdiagnosis was greater in individuals aged older than 64 years, men and in some types of cancers (skin, colorectal, prostate). Although the reduction was predominantly focused during the lockdown, expected figures have not yet been reached (40.5% reduction during the lockdown and 24.3% reduction after that)., Conclusions: Reduction in cancer incidence has been observed during and after the lockdown. Urgent policy interventions are necessary to mitigate the indirect effects of the COVID-19 pandemic and related control measures on other diseases and some strategies must be designed in order to reduce the underdiagnosis of cancer., Competing Interests: Competing interests: None declared., (© Author(s) (or their employer(s)) 2021. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.)

Polyphenols and omega-3 polyunsaturated fatty acids from fish oils, i.e., eicosapentaenoic and docosahexaenoic acids, are well-recognized nutraceuticals, and their single antioxidant and anti-inflammatory properties have been demonstrated in several studies found in the literature. It has been reported that the combination of these nutraceuticals can lead to three-fold increases in glutathione peroxidase activity, two-fold increases in plasma antioxidant capacity, decreases of 50-100% in lipid peroxidation, protein carbonylation, and urinary 8-isoprotanes, as well as 50-200% attenuation of common inflammation biomarkers, among other effects, as compared to their individual capacities. Therefore, the adequate combination of those bioactive food compounds and their single properties should offer a powerful tool for the design of successfully nutritional interventions for the prevention and palliation of a plethora of human metabolic diseases, frequently diet-induced, whose etiology and progression are characterized by redox homeostasis disturbances and a low-grade of chronic inflammation. However, the certain mechanisms behind their biological activities, in vivo interaction (both between them and other food compounds), and their optimal doses and consumption are not well-known yet. Therefore, we review here the recent evidence accumulated during the last decade about the cooperative action between polyphenols and fish oils against diet-related metabolic alterations, focusing on the mechanisms and pathways described and the effects reported. The final objective is to provide useful information for strategies for personalized nutrition based on these nutraceuticals.

Adipose tissue is now recognized as an active organ with an important homeostatic function in glucose and lipid metabolism and the development of insulin resistance. The present research investigates the role of lipid mediators and lipid profiling for controlling inflammation and the metabolic normal function of white adipose tissue from rats suffering from diet-induced prediabetes. Additionally, the contribution to the adipose lipidome induced by the consumption of marine ω-3 PUFAs as potential regulators of inflammation is addressed. For that, the effects on the inflammatory response triggered by high-fat high-sucrose (HFHS) diets were studied in male Sprague-Dawley rats. Using SPE-LC-MS/MS-based metabolo-lipidomics, a range of eicosanoids, docosanoids and specialized pro-resolving mediators (SPMs) were measured in white adipose tissue. The inflammatory response occurring in prediabetic adipose tissue was associated with the decomposition of ARA epoxides to ARA-dihydroxides, the reduction of oxo-derivatives and the formation of prostaglandins (PGs). In an attempt to control the inflammatory response initiated, LOX and non-enzymatic oxidation shifted toward the production of the less pro-inflammatory EPA and DHA metabolites rather than the high pro-inflammatory ARA hydroxides. Additionally, the change in LOX activity induced the production of intermediate hydroxides precursors of SPMs as protectins (PDs), resolvins (Rvs) and maresins (MaRs). This compensatory mechanism to achieve the restoration of tissue homeostasis was significantly strengthened through supplementation with fish oils. Increasing proportions of ω-3 PUFAs in adipose tissue significantly stimulated the formation of DHA-epoxides by cytochrome P450, the production of non-enzymatic EPA-metabolites and prompted the activity of 12LOX. Finally, protectin PDX was significantly reduced in the adipose tissue of prediabetic rats and highly enhanced through ω-3 PUFAs supplementation. Taken together, these actively coordinated modifications constitute key mechanisms to restore adipose tissue homeostasis with an important role of lipid mediators. This compensatory mechanism is reinforced through the supplementation of the diet with fish oils with high and balanced contents of EPA and DHA. The study highlights new insides on the targets for effective treatment of incipient diet-induced diabetes and the mechanism underlying the potential anti-inflammatory action of marine lipids., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The reviewer AG-I declared a shared affiliation with several of the authors (IM, GD, LM, JT and SR) to the handling editor at the time of the review., (Copyright © 2021 Dasilva, Lois, Méndez, Miralles-Pérez, Romeu, Ramos-Romero, Torres and Medina.)

The aim of the present study was to evaluate the influence of several experimental pretreatment crosslinker solutions on the resin polymer-dentine interface created using a representative universal adhesive system, by means of microtensile bond strength testing (μTBS), nanomechanical properties and ultramorphology confocal laser scanning microscopy (CLSM). Five experimental solutions containing different flavonoids were applied as dentine pretreatment after acid etching. A control pretreatment group containing no flavonoid was also employed. A representative modern universal adhesive was then applied, followed by a 3 mm thick composite built up. Specimens were sectioned into sticks and submitted to a μTBS test or nanoindentation analysis along the interface (24 h or 25,000 thermocycles). The ultramorphology of the polymer-resin interface was also evaluated using CLSM. The results were analyzed using two-way ANOVA and Bonferroni's post hoc test (α = 0.05). All flavonoids improved short- and long-term μTBS values ( p < 0.01), while only some specific such solutions improved the nanomechanical properties ( p < 0.05) and preserved the structural morphology of the interface after aging. Pretreatment of acid-etched dentine using specific flavonoid-containing solutions may be a promising approach to improve both the nanomechanical properties and the durability of modern universal adhesive systems.

The Lactobacillus plantarum and Lactobacillus pentosus genotypes existing in industrial-scale cucumber fermentations were defined using rep-PCR-(GTG)5. The ability of each genotype to ferment cucumbers under various conditions was evaluated. Rep-PCR-(GTG)5 was the technique capable of illustrating the most intraspecies discrimination compared to the sequencing of housekeeping genes (recA, dnaK, pheS and rpoA), MLST and RAPD with primers LP1, OPL5, M14 and COC. Ten genotypic clusters were defined for the 199 L. pentosus tested and three for the 17 L. plantarum clones. The ability of the 216 clones genotyped and 37 additional cucumber fermentation isolates, of the same species, to rapidly decrease the pH of cucumber juice medium under various combinations of sodium chloride (0 or 6%), initial pH (4.0 or 5.2) and temperatures (15 or 30 °C) was determined using a fractional factorial screening design. A reduced fermentation ability was observed for the L. plantarum strains as compared to L. pentosus, except for clone 3.2.8, which had a ropy phenotype and aligned to genotypic cluster A. L. pentosus strains belonging to three genotypic clusters (B, D and J) were more efficient in cucumber juice fermentation as compared to most L. plantarum strains. This research identified three genetically diverse L. pentosus strains and one L. plantarum as candidates for starter cultures for commercial cucumber fermentations., (Copyright © 2020. Published by Elsevier Ltd.)

Background: There is evidence that an ongoing patient-physician relationship is associated with improved health outcomes and more efficient health systems. The main objective of this study is to describe the continuity of care in primary healthcare in Catalonia (Spain) and to analyze whether the organization of primary care practices (PCP) or their patients' sociodemographic characteristics play a role in its continuity of care., Methods: Four indices were used to measure continuity of care: Usual Provider Index (UPC), Modified Modified Continuity Index (MMCI), Continuity of Care Index (COC), and Sequential Continuity Index (SECON). The study was conducted on 287 PCP of the Catalan Institute of Health (Institut Català de la Salut-ICS). Each continuity of care index was calculated at the patient level (3.2 million patients and 35.5 million visits) and then aggregated at the PCP level. We adjusted linear regression models for each continuity index studied, considering the result of the index as an independent variable and demographic and organizational characteristics of the PCP as explanatory variables. Pearson correlation tests were used to compare the four continuity of care indices., Results: Indices' results were: UPC: 70,5%; MMCI: 73%; COC: 53,7%; SECON: 60,5%. The continuity of care indices had the highest bivariate correlation with the percentage of appointments booked with an assigned health provider (VISUBA variable: the lower the value, the higher the visits without an assigned health provider, and thus an organization favoring immediate consultation). Its R 2 ranged between 56 and 63%, depending on the index. The multivariate model which explained better the variability of continuity of care indices (from 49 to 56%) included the variables VISUBA and rurality with a direct relationship; while the variables primary care physician leave days and training practices showed an inverse relationship., Conclusion: Study results suggest that an organization of primary care favoring immediate consultation is related to a lower continuity of patient care.

Background: Pneumonia is one of the complications of COVID-19. Primary care electronic health records (EHR) have shown the utility as a surveillance system. We therefore analyse the trends of pneumonia during two waves of COVID-19 pandemic in order to use it as a clinical surveillance system and an early indicator of severity., Methods: Time series analysis of pneumonia cases, from January 2014 to December 2020. We collected pneumonia diagnoses from primary care EHR, a software system covering > 6 million people in Catalonia (Spain). We compared the trend of pneumonia in the season 2019-2020 with that in the previous years. We estimated the expected pneumonia cases with data from 2014 to 2018 using a time series regression adjusted by seasonality and influenza epidemics., Results: Between 4 March and 5 May 2020, 11,704 excess pneumonia cases (95% CI: 9909 to 13,498) were identified. Previously, we identified an excess from January to March 2020 in the population older than 15 years of 20%. We observed another excess pneumonia period from 22 october to 15 november of 1377 excess cases (95% CI: 665 to 2089). In contrast, we observed two great periods with reductions of pneumonia cases in children, accounting for 131 days and 3534 less pneumonia cases (95% CI, 1005 to 6064) from March to July; and 54 days and 1960 less pneumonia cases (95% CI 917 to 3002) from October to December., Conclusions: Diagnoses of pneumonia from the EHR could be used as an early and low cost surveillance system to monitor the spread of COVID-19.

The combined supplementation of buckwheat D-fagomine (FG) and fish omega-3 polyunsaturated fatty acids (ω-3 PUFA) attenuates the development of insulin resistance in rats fed a high-fat (HF) diet. This study aimed to examine the effects of combined supplementation with FG and ω-3 PUFA on dyslipidemia, transaminases, interleukin-6, and oxidative stress. Forty-five male Sprague-Dawley rats were fed a standard diet, an HF diet, an HF diet supplemented with FG, an HF diet supplemented with ω-3 PUFA, or an HF diet supplemented with FG and ω-3 PUFA for 21 weeks. Triacylglycerol, cholesterol, aspartate aminotransferase, alanine aminotransferase, and interleukin-6 were measured. The assessment of oxidative stress included plasma antioxidant capacity, antioxidant enzyme activities, glutathione content, lipid peroxidation, and protein carbonylation. The combined supplementation with FG and ω-3 PUFA did not attenuate the slight accumulation of liver cholesterol induced by the HF diet but normalized the plasma alanine aminotransferase activity. Rats fed the HF diet supplemented with the combination showed a lower amount of plasma interleukin-6 than those fed a standard diet. The combination attenuated oxidative damage induced by the HF diet, decreased antioxidant enzyme activities, and enhanced glutathione status. The beneficial effects of the combination of FG and ω-3 PUFA on oxidative stress and related risk factors in pre-obese rats were mainly modulated by ω-3 PUFA.

The constitutively active tyrosine-kinase BCR/ABL1 oncogene plays a key role in human chronic myeloid leukemia development and disease maintenance, and determines most of the features of this leukemia. For this reason, tyrosine-kinase inhibitors are the first-line treatment, offering most patients a life expectancy like that of an equivalent healthy person. However, since the oncogene stays intact, lifelong oral medication is essential, even though this triggers adverse effects in many patients. Furthermore, leukemic stem cells remain quiescent and resistance is observed in approximately 25% of patients. Thus, new therapeutic alternatives are still needed. In this scenario, the interruption/deletion of the oncogenic sequence might be an effective therapeutic option. The emergence of CRISPR (clustered regularly interspaced short palindromic repeats) technology can offer a definitive treatment based on its capacity to induce a specific DNA double strand break. Besides, it has the advantage of providing complete and permanent oncogene knockout, while tyrosine kinase inhibitors (TKIs) only ensure that BCR-ABL1 oncoprotein is inactivated during treatment. CRISPR/Cas9 cuts DNA in a sequence-specific manner making it possible to turn oncogenes off in a way that was not previously feasible in humans. This review describes chronic myeloid leukemia (CML) disease and the main advances in the genome-editing field by which it may be treated in the future.

Carbonylation is a nonenzymatic irreversible posttranslational protein modification and the main hallmark of protein oxidative damage. Elevated levels of protein carbonyl groups have been detected in age-related and metabolic diseases such as obesity, diabetes, Alzheimer, Parkinson, and several other oxidative stress-related maladies. Interestingly, many studies have shown that only a subset of proteins is carbonylated under the conditions of oxidative stress, demonstrating that carbonylation is a highly selective process. As a consequence, identifying and quantifying the disease-induced changes on a certain carbonylome are crucial to understanding the etiology and progression of numerous diseases and then designing adequate prevention/palliation strategies. However, the low abundance of carbonylated proteins in vivo, the enormous diversity of reactive species, and their relative lability make the analysis of carbonylated proteins a challenging task for redox proteomic technology. Therefore, we present a proteomic approach based on the labeling of carbonyls formed in vivo on proteins using the fluorescein 5-thiosemicarbazide (FTSC) tag to detect the subset of carbonylated proteins among a complex mixture of proteins regardless of the nature of carbonyl adduct, isolation and relative quantification of carbonylated proteins in 2D gel electrophoresis, and protein identification by LC-MS/MS analysis. This method has been successfully used for the evaluation of in vivo protein carbonylation in very diverse animal tissues (plasma, liver, kidney, skeletal muscle, and adipose tissue) and species (from fish to mammalian) and has also been applied in different research fields (from food technology to nutrition), demonstrating its robustness and reliability.

8 March (8M), now known as International Women's Day, is a day for feminist claims where demonstrations are organized in over 150 countries, with the participation of millions of women all around the world. These demonstrations can be viewed as collective rituals and thus focus attention on the processes that facilitate different psychosocial effects. This work aims to explore the mechanisms (i.e., behavioral and attentional synchrony, perceived emotional synchrony, and positive and transcendent emotions) involved in participation in the demonstrations of 8 March 2020, collective and ritualized feminist actions, and their correlates associated with personal well-being (i.e., affective well-being and beliefs of personal growth) and collective well-being (i.e., social integration variables: situated identity, solidarity and fusion), collective efficacy and collective growth, and behavioral intention to support the fight for women's rights. To this end, a cross-cultural study was conducted with the participation of 2,854 people (age 18-79; M = 30.55; SD = 11.66) from countries in Latin America (Mexico, Chile, Argentina, Brazil, Peru, Colombia, and Ecuador) and Europe (Spain and Portugal), with a retrospective correlational cross-sectional design and a convenience sample. Participants were divided between demonstration participants ( n = 1,271; 94.0% female) and non-demonstrators or followers who monitored participants through the media and social networks ( n = 1,583; 75.87% female). Compared with non-demonstrators and with males, female and non-binary gender respondents had greater scores in mechanisms and criterion variables. Further random-effects model meta-analyses revealed that the perceived emotional synchrony was consistently associated with more proximal mechanisms, as well as with criterion variables. Finally, sequential moderation analyses showed that proposed mechanisms successfully mediated the effects of participation on every criterion variable. These results indicate that participation in 8M marches and demonstrations can be analyzed through the literature on collective rituals. As such, collective participation implies positive outcomes both individually and collectively, which are further reinforced through key psychological mechanisms, in line with a Durkheimian approach to collective rituals., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2020 Zumeta, Castro-Abril, Méndez, Pizarro, Włodarczyk, Basabe, Navarro-Carrillo, Padoan-De Luca, da Costa, Alonso-Arbiol, Torres-Gómez, Cakal, Delfino, Techio, Alzugaray, Bilbao, Villagrán, López-López, Ruiz-Pérez, Cedeño, Reyes-Valenzuela, Alfaro-Beracoechea, Contreras-Ibáñez, Ibarra, Reyes-Sosa, Cueto, Carvalho and Pinto.)

Background: To analyse the impact of the COVID-19 epidemic and the lockdown measures on the follow-up and control of chronic diseases in primary care., Methods: Retrospective study in 288 primary care practices (PCP) of the Catalan Institute of Health. We analysed the results of 34 indicators of the Healthcare quality standard (EQA), comprising different types: treatment (4), follow-up (5), control (10), screening (7), vaccinations (4) and quaternary prevention (4). For each PCP, we calculated each indicator's percentage of change in February, March and April 2020 respective to the results of the previous month; and used the T-Student test for paired data to compare them with the percentage of change in the same month of the previous year. We defined indicators with a negative effect those with a greater negative change or a lesser positive change in 2020 in comparison to 2019; and indicators with a positive effect those with a greater positive change or a lesser negative change., Results: We observed a negative effect on 85% of the EQA indicators in March and 68% in April. 90% of the control indicators had a negative effect, highlighting the control of LDL cholesterol with a reduction of - 2.69% (95%CI - 3.17% to - 2.23%) in March and - 3.41% (95%CI - 3.82% to - 3.01%) in April; and the control of blood pressure with a reduction of - 2.13% (95%CI - 2.34% to - 1.9%) and - 2.59% (95%CI - 2.8% to - 2.37%). The indicators with the greatest negative effect were those of screening, such as the indicator of diabetic foot screening with a negative effect of - 2.86% (95%CI - 3.33% to - 2.39%) and - 4.13% (95%CI - 4.55% to - 3.71%) in March and April, respectively. Only one vaccination indicator, adult Measles-Mumps-Rubella vaccine, had a negative effect in both months. Finally, among the indicators of quaternary prevention, we observed negative effects in March and April although in that case a lower inadequacy that means better clinical outcome., Conclusions: The COVID-19 epidemic and the lockdown measures have significantly reduced the results of the follow-up, control, screening and vaccination indicators for patients in primary care. On the other hand, the indicators for quaternary prevention have been strengthened and their results have improved.

Aquaporins (AQPs) facilitate water and glycerol movement across membranes. AQP7 is the main aquaglyceroporin in pancreatic β-cells and was proposed to play a role in insulin exocytosis. Although AQP7-null mice display adult-onset obesity, impaired insulin secretion and insulin resistance, AQP7 loss-of-function homozygous mutations in humans do not correlate with obesity nor type-2 diabetes. In addition, AQP12 is upregulated in pancreatitis. However, the implication of this isoform in endocrine pancreas inflammation is still unclear. Here, we investigated AQP7 and AQP12 involvement in cellular and inflammatory processes using RIN-m5F beta cells, a model widely used for their high insulin secretion. AQP7 and AQP12 expression were directly associated with cell proliferation, adhesion and migration. While tumor necrosis factor-alpha (TNFα)-induced inflammation impaired AQP7 expression and drastically reduced insulin secretion, lipopolysaccharides (LPS) prompted AQP7 upregulation, and both TNFα and LPS upregulated AQP12. Importantly, cells overexpressing AQP12 are more resistant to inflammation, revealing lower levels of proinflammatory markers. Altogether, these data document AQP7 involvement in insulin secretion and AQP12 implication in inflammation, highlighting their fundamental role in pancreatic β-cell function., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Collisions of He 2+ , Li 3+ and C 3+ ions with water molecules are studied at energies ranging between 20 keV u -1 and 500 keV u -1 . Three methods are employed: the classical trajectory Monte Carlo (CTMC), the expansion of the scattering wave function in terms of asymptotic frozen molecular orbitals (AFMO) and a lattice method to numerically solve the time-dependent Schrödinger equation (GridTDSE). Total cross sections for single ionization, single electron capture, transfer ionization and electron production are calculated and compared with previous close-coupling calculations and experiments. The fragmentation branching ratios are discussed.

The aim of the present study was to perform a screening of extracts obtained from 15 medicinal plants using water (at 25 and 90 °C) or ethanol (at 25 °C), to bactericidal activity against cariogenic S. mutans ATCC 25175, as well as to carry out the preliminary phytochemical characterization of the extracts and HPLC/MS assay for selected extracts. The extractions were carried out for 5 h at 400 rpm. Only five from 45 tested extracts were selected based on their antibacterial activity. The IC 50 varied from of 28 ppm for Quercus ilex up to 250 ppm for Jatropha dioica . Different polyphenolic and quinic acids, flavonoids, anthocyanin or tyrosol were identified by HPLC/MS in selected extracts from Rosa gallica L. , Jatropha dioica Sessé , Mimosa tenuiflora (Willd.) Poir , Quercus ilex L., and Solanum nigrum . The obtained results confirm that selected extracts are good candidates to be used for cariogenic bacteria control.

There is scarce investigation addressing interpopulation tolerance responses to address the influence of a history of chronic stress exposure, as that occurring in polluted environments, in photoautotrophs. We evaluated ecophysiological (photosynthetic activity) and metabolic (oxidative stress and damage) responses of two populations of green macroalga Ulva compressa from polluted (Ventanas) and non-polluted (Cachagua) localions of central Chile, and exposed to controlled hypersalinity conditions of 32 (control), 42, 62 and 82 psu (practical salinity units) for 6 h, 48 h and 6 d. Both primary production (ETR max ) and photosynthetic efficiency (α ETR ) were generally higher in the population from Cachagua compared to Ventanas at all times and salinities. Moreover, at most experimental times and salinities the population from Ventanas had greater levels of H 2 O 2 and lipid peroxidation that individuals from Cachagua. Total ascorbate was higher in the population of Cachagua than Ventanas at 42 and 82 psu after 6 and 48 h, respectively, while at 6 d concentrations were similar between both populations at all salinities. Total glutathione was greater in both populations after 6 h at all salinities, but at 48 h its concentrations were higher only in the population from Cachagua, a trend that was maintained at 6 d under 82 psu only. Reduced and oxidized ascorbate (ASC and DHA, respectively) and glutathione (GSH and GSSG, respectively) demonstrated similar patterns between U. compressa populations, with an increase oxidation with greater salinities but efficient recycling to maintain sufficient batch of ASC and GSH. When assessing the expression of antioxidant enzymes catalase (CAT), superoxide dismutase (SOD) and dehydroascorbate reductase (DHAR), while the population of Ventanas displayed a general trend of upregulation with increasing salinities along the experiments, U. compressa from Cachagua revealed patterns of downregulation. Results demonstrated that although both populations were still viable after the applied hypersalinities during all experimental times, biological performance was usually more affected in the population from the Ventanas than Cachagua, likely due to a depressed baseline metabolism after a long history of exposition to environmental pollution., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Ticks and the vast array of pathogens they transmit, including bacteria, viruses, protozoa, and helminths, constitute a growing burden for human and animal health worldwide. In Cuba, the major tropical island in the Caribbean, ticks are an important cause of vector-borne diseases affecting livestock production, pet animal health and, to a lesser extent, human health. The higher number of tick species in the country belong to the Argasidae family and, probably less known, is the presence of an autochthonous tick species in the island, Ixodes capromydis . Herein, we provide a comprehensive review of the ticks and tick-borne pathogens (TBPs) affecting animal and human health in Cuba. The review covers research results including ecophysiology of ticks, the epidemiology of TBPs, and the diagnostic tools used currently in the country for the surveillance of TBPs. We also introduce the programs implemented in the country for tick control and the biotechnology research applied to the development of anti-tick vaccines., Competing Interests: The authors declare no conflict of interest.

A synthetic 20 amino acid peptide of the ribosomal protein P0 from ticks, when conjugated to keyhole limpet hemocyanin from Megathura crenulata and used as an immunogen against Rhipicephalus microplus and Rhipicephalus sanguineus s.l. species, has shown efficacies of around 90%. There is also experimental evidence of a high efficacy of this conjugate against Amblyomma mixtum and Ixodes ricinus species, which suggest that this antigen could be a good broad-spectrum anti-tick vaccine candidate. In this study, the P0 peptide (pP0) was chemically conjugated to Bm86 as a carrier protein. SDS-PAGE analysis of this conjugate demonstrated that it is highly heterogeneous in size, carrying from 1 to 18 molecules of pP0 per molecule of Bm86. Forty-nine out of the 54 lysine residues and the N-terminal end of Bm86 were found partially linked to pP0 by using LC-MS/MS analysis and the combination of four different softwares. Several post-translational modifications of Bm86 protein were also identified by mass spectrometry. High immunogenicity and efficacy were achieved when dogs and cattle were vaccinated with the pP0-Bm86 conjugate and challenged with R. sanguineus s.l. and R. microplus , respectively. These results encourage the development of this antigen with promising possibilities as an anti-tick vaccine.

Diacylglycerols (DAG) and ceramides have been suggested as early predictors of insulin resistance. This study was aimed to examine the combined effects of fish oil (FO) and grape seed extract (GSE) on hepatic endogenous antioxidants, DAG and ceramides in diet-induced early stages of insulin resistance. Thirty-five rats were fed one of the following diets: (1) a standard diet (STD group), (2) a high-fat high-sucrose diet (HFHS group), (3) an HFHS diet enriched with FO (FO group), (4) an HFHS diet enriched with GSE (GSE group) or (5) an HFHS diet enriched with FO and GSE (FO + GSE group). In the liver, endogenous antioxidants were measured using spectrophotometric and fluorometric techniques, and non-targeted lipidomics was conducted for the assessment of DAG and ceramides. After 24 weeks, the FO + GSE group showed increased glutathione peroxidase activity, as well as monounsaturated fatty acid and polyunsaturated fatty acid-containing DAG, and long-chain fatty acid-containing ceramides abundances compared to the STD group. The FO and GSE combination induced similar activation of the antioxidant system and bioactive lipid accumulation in the liver than the HFHS diet without supplementation. In addition, the FO and GSE combination increased the abundances of polyunsaturated fatty acid-containing DAG in the liver.

Aquaporin-11 (AQP11) is expressed in human adipocytes, but its functional role remains unknown. Since AQP11 is an endoplasmic reticulum (ER)-resident protein that transports water, glycerol, and hydrogen peroxide (H 2 O 2 ), we hypothesized that this superaquaporin is involved in ER stress induced by lipotoxicity and inflammation in human obesity. AQP11 expression was assessed in 67 paired visceral and subcutaneous adipose tissue samples obtained from patients with morbid obesity and normal-weight individuals. We found that obesity and obesity-associated type 2 diabetes increased ( p < 0.05) AQP11 mRNA and protein in visceral adipose tissue, but not subcutaneous fat. Accordingly, AQP11 mRNA was upregulated ( p < 0.05) during adipocyte differentiation and lipolysis, two biological processes altered in the obese state. Subcellular fractionation and confocal microscopy studies confirmed its presence in the ER plasma membrane of visceral adipocytes. Proinflammatory factors TNF-α, and particularly TGF-β1, downregulated ( p < 0.05) AQP11 mRNA and protein expression and reinforced its subcellular distribution surrounding lipid droplets. Importantly, the AQP11 gene knockdown increased ( p < 0.05) basal and TGF-β1-induced expression of the ER markers ATF4 and CHOP. Together, the downregulation of AQP11 aggravates TGF-β1-induced ER stress in visceral adipocytes. Owing to its "peroxiporin" properties, AQP11 overexpression in visceral fat might constitute a compensatory mechanism to alleviate ER stress in obesity.

This study aimed to express heterologously the lipase LipA from Pseudomonas aeruginosa PSA01 obtained from palm fruit residues. In previous approaches, LipA was expressed in Escherichia coli fused with its signal peptide and without its disulfide bond, displaying low activity. We cloned the mature LipA with its truncated chaperone Lif in a dual plasmid and overexpressed the enzyme in two E. coli strains: the traditional BL21 (DE3) and the SHuffle ® strain, engineered to produce stable cytoplasmic disulfide bonds. We evaluated the effect of the disulfide bond on LipA stability using molecular dynamics. We expressed LipA successfully under isopropyl β-d-1-thio-galactopyranoside (IPTG) and slow autoinducing conditions. The SHuffle LipA showed higher residual activity at 45 °C and a greater hyperactivation after incubation with ethanol than the enzyme produced by E. coli BL21 (DE3). Conversely, the latter was slightly more stable in methanol 50% and 60% (t½: 49.5 min and 9 min) than the SHuffle LipA (t½: 31.5 min and 7.4 min). The molecular dynamics simulations showed that removing the disulfide bond caused some regions of LipA to become less flexible and some others to become more flexible, significantly affecting the closing lid and partially exposing the active site at all times., Competing Interests: The authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analysis, or interpretation of data; in the writing of the manuscript; or in the decision to publish the results.

Proteasome inhibitors have great success for their therapeutic potential against hematologic malignancies. First generation proteasome inhibitor bortezomib induced peripheral neuropathy is considered as a limiting factor in chemotherapy and its second-generation counterpart carfilzomib is associated with lower rates of neurotoxicity. The mitochondrial toxicity (mitotoxicity) hypothesis arises from studies with animal models of bortezomib induced peripheral neuropathy. However, molecular mechanisms are not fully elucidated and the role of mitotoxicity in bortezomib and carfilzomib induced neurotoxicity has not been investigated comparatively. Herein, we characterized the neurotoxic effects of bortezomib and carfilzomib at the molecular level in human neuronal cells using LC-MS/MS analysis, flow cytometry, RT-qPCR, confocal microscopy and western blotting. We showed that bortezomib and carfilzomib affected the human neuronal proteome differently, and bortezomib caused higher proteotoxic stress via protein oxidation, protein K48-ubiquitination, heat shock protein expression upregulation and reduction of mitochondria membrane potential. Bortezomib and carfilzomib did not affect the gene expression levels related to mitochondrial dynamics (optic atrophy 1; OPA1, mitofusin 1; MFN1, mitofusin 2; MFN2, fission 1; FIS1, dynamin-related protein 1; DRP1) and overall mitophagy rate whereas, PINK1/Parkin mediated mitophagy gene expressions were altered with both drugs. Bortezomib and carfilzomib caused downregulation of the contents of mitochondrial oxidative phosphorylation complexes, voltage-dependent anion channel 1 (VDAC1) and uncoupling protein 2 (UCP2) similarly. Our findings suggest that, both drugs induce mitotoxicity besides proteotoxic stress in human neuronal cells and the higher incidence of neurotoxicity with bortezomib than carfilzomib is not directly related to mitochondrial pathways., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)

Biological communities change in response to human alteration. The response of individual taxa and the community can be used to establish preventive criteria to halt further biodiversity deterioration. Here we explore how consistent are the boundaries between Good and Moderate ecological status derived from classification systems used in North-NW Spain: NORThern Spain Indicators system (NORTI), River type specific multimetric (METI) and Iberian Bio-monitoring Working Party (IBMWP), by using common interpretation of normative definitions of Water Framework Directive. We applied the three classifications to a monitoring dataset of Nalón River basin, comprising samples from different stream types and reference conditions. We applied Threshold Indicator Taxa ANalysis to the invertebrate community along the most relevant environmental pressures and biological impairment gradients represented by the Ecological Quality Ratio (EQR) scores of the classification systems. Only NORTI provided a true community ecological threshold and the change point (cp) 95% quantile (Q95%) range of 0.760 was assumed to be the boundary from Good to Moderate (G/M) status, used to standardize the number of taxa loss in all systems. Since the average number of taxa at reference sites was 34, the estimated loss of sensitive taxa was up to 97.1% in IBMWP, 73.5% in METI and 52.9% in NORTI when passing from Good to Moderate status, revealing very permissive boundaries. The loss of common sensitive taxa in NORTI at Q95% was used as G/M threshold and applied to the other classifications, resulting all in a common biodiversity loss of 21% of sensitive taxa richness at values of NORTI-EQR = 0.760, METI-EQR = 0.818 and IBMWP-EQR = 0.753. Results indicate that significant community changes along pressure gradients allow for establishing quantitative criteria consistent with normative definitions. This understanding derived from Directive monitoring programs can assess the risk that invertebrate communities face in terms of species loss derived from anthropogenic pressures., Competing Interests: Declaration of competing interest None., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background/objectives: Bariatric surgery improves nonalcoholic fatty liver disease (NAFLD) and nonalcoholic steatohepatitis (NASH), but the underlying mechanisms remain elusive. We evaluated the potential role of ghrelin isoforms in the amelioration of hepatic inflammation after sleeve gastrectomy and Roux-en-Y gastric bypass (RYGB)., Subjects/methods: Plasma ghrelin isoforms were measured in male Wistar rats (n = 129) subjected to surgical (sham operation, sleeve gastrectomy, or RYGB) or dietary interventions [fed ad libitum a normal (ND) or a high-fat diet (HFD) or pair-fed diet]. The effect of acylated and desacyl ghrelin on markers of inflammation, mitochondrial dysfunction, and endoplasmic reticulum (ER) stress in primary rat hepatocytes under palmitate-induced lipotoxic conditions was assessed., Results: Plasma desacyl ghrelin was decreased after sleeve gastrectomy and RYGB, whereas the acylated/desacyl ghrelin ratio was augmented. Both surgeries diminished obesity-associated hepatic steatosis, CD68 + - and apoptotic cells, proinflammatory JNK activation, and Crp, Tnf, and Il6 transcripts. Moreover, a postsurgical amelioration in the mitochondrial DNA content, oxidative phosphorylation (OXPHOS) complexes I and II, and ER stress markers was observed. Specifically, following bariatric surgery GRP78, spliced XBP-1, ATF4, and CHOP levels were reduced, as were phosphorylated eIF2α. Interestingly, acylated and desacyl ghrelin inhibited steatosis and inflammation of palmitate-treated hepatocytes in parallel to an upregulation of OXPHOS complexes II, III, and V, and a downregulation of ER stress transducers IRE1α, PERK, ATF6, their downstream effectors, ATF4 and CHOP, as well as chaperone GRP78., Conclusions: Our data suggest that the increased relative acylated ghrelin levels after bariatric surgery might contribute to mitigate obesity-associated hepatic inflammation, mitochondrial dysfunction, and ER stress.

The Nomo1 gene mediates a wide range of biological processes of importance in embryonic development. Accordingly, constitutive perturbation of Nomo1 function may result in myriad developmental defects that trigger embryonic lethality. To extend our understanding of Nomo1 function in postnatal stages and in a tissue-specific manner, we generated a conditional knockout mouse model of Nomo1. To achieve this, we used clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 technology in C57Bl/6J mouse zygotes to generate a new mouse model in which exon 3 of the Nomo1 gene is specifically flanked (or floxed) by LoxP sites (Nomo1 f/f ). Nomo1 f/f mouse embryonic fibroblasts were transduced with a Cre adenovirus and efficiently recombined between LoxP sites. Genomic and expression studies in Nomo1-transduced MEFs demonstrated that the Nomo1 exon 3 is ablated. Western blot assay showed that no protein or early truncated protein is produced. In vivo assay crossing Nomo1 f/f mouse with a Msi1-CRE transgenic mouse corroborated the previous findings and it showed Nomo1 exon 3 deletion at msi1+ cell compartment. This short technical report demonstrates that CRISPR/Cas9 technology is a simple and easy method for creating conditional mouse models. The Nomo1 f/f mouse will be useful to researchers who wish to explore the role of Nomo1 in any developmental stage or in a tissue-specific manner.

Hydrogen peroxide (H 2 O 2 ) is generated in cells and plays an important role as a signalling molecule. It has been reported that H 2 O 2 is involved in physiological and pathological processes in skeletal muscle. However, H 2 O 2 detection in cells with traditional techniques produces frequent artefacts. Currently, the HyPer biosensor detects intracellular H 2 O 2 specifically in real time using fluorescence microscopy. The aim of this study was to develop and optimize approaches used to express the HyPer biosensor in different models of skeletal muscle cells, such as the C2C12 myoblast/myotube cell line and mature skeletal muscle fibres isolated from C57BL/6J mice, and to measure intracellular H 2 O 2 in real time in these cells. The results show that the expression of the HyPer biosensor in skeletal muscle cells is possible. In addition, we demonstrate that HyPer is functional and that this biosensor detects changes and fluctuations in intracellular H 2 O 2 in a reversible manner. The HyPer2 biosensor, which is a more advanced version of HyPer, presents improved properties in terms of sensitivity in detecting lower concentrations of H 2 O 2 in skeletal muscle fibres. In conclusion, the expression of the HyPer biosensor in the different experimental models combined with fluorescence microscopy techniques is a powerful methodology to monitor and register intracellular H 2 O 2 specifically in skeletal muscle. The innovation of the methodological approaches presented in this study may present new avenues for studying the role of H 2 O 2 in skeletal muscle pathophysiology. Furthermore, the methodology may potentially be adapted to yield other specific biosensors for different reactive oxygen and nitrogen species or metabolites involved in cellular functions.

Despite the concern about anthropogenic heavy metal accumulation, there remain few multi-level ecotoxicological studies to evaluate their effects in fluvial ecosystems. The toxicity of field-collected sediments exhibiting a gradient of heavy metal contamination (Cd, Pb, and Zn) was assessed in Chironomus riparius. For this purpose, larvae were exposed throughout their entire life cycle to these sediments, and toxic effects were measured at different levels of biological organization, from the molecular (lipidomic analysis and transcriptional profile) to the whole organism response (respiration rate, shape markers, and emergence rate). Alterations in the activity of relevant genes, as well as an increase of storage lipids and decrease in membrane fluidity, were detected in larvae exposed to the most contaminated sediments. Moreover, reduced larval and adult mass, decrease of larval respiration rate, and delayed emergence were observed, along with increased mentum and mandible size in larvae and decreased wing loading in adults. This study points out the deleterious effects of heavy metal exposure at various levels of biological organization and provides some clues regarding the mode of toxic action. This integrative approach provides new insights into the multi-level effects on aquatic insects exposed to heavy metal mixtures in field sediments, providing useful tools for ecological risk assessment in freshwater ecosystems., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Diet-induced obesity has been linked to metabolic disorders such as cardiovascular diseases andtype 2 diabetes. A factor linking diet to metabolic disorders is oxidative stress, which can damagebiomolecules, especially proteins. The present study was designed to investigate the effect of marineomega-3 polyunsaturated fatty acids (PUFAs) (eicosapentaenoic acid (EPA) and docosahexaenoic acid(DHA)) and their combination with grape seed polyphenols (GSE) on carbonyl-modified proteins fromplasma and liver in Wistar Kyoto rats fed an obesogenic diet, namely high-fat and high-sucrose (HFHS)diet. A proteomics approach consisting of fluorescein 5-thiosemicarbazide (FTSC) labelling of proteincarbonyls, visualization of FTSC-labelled protein on 1-DE or 2-DE gels, and protein identification byMS/MS was used for the protein oxidation assessment. Results showed the efficiency of the combinationof both bioactive compounds in decreasing the total protein carbonylation induced by HFHS diet in bothplasma and liver. The analysis of carbonylated protein targets, also referred to as the 'carbonylome',revealed an individual response of liver proteins to supplements and a modulatory effect on specificmetabolic pathways and processes due to, at least in part, the control exerted by the supplements on theliver protein carbonylome. This investigation highlights the additive effect of dietary fish oils and grapeseed polyphenols in modulating in vivo oxidative damage of proteins induced by the consumption ofHFHS diets., Competing Interests: The authors declare no conflict of interest.

Recycling sewage sludge by applying it to agricultural land is strategically important in the European Union and is regulated by Directive 86/278/EEC, aimed at protecting the soil and humans from the presence of unwanted substances. However, because of the ruminant feeding habits, there is a risk that animals grazed on pasture or fed crops grown on land treated in this way may ingest biosolids adhered to foliage and/or on the top soil. This paper describes an episode of toxicity in a dairy herd consuming silage from a field fertilized with sewage sludge produced in a wastewater treatment plant. The affected cows were recumbent, unable to rise and suffered diarrhoea. Analysis of tissues (fresh weight) from a cow that subsequently died revealed severe hepatic iron loading (6720 mg/kg) and secondary multi-trace element hepatic deficiency, particularly of copper (0.812 mg/kg) and manganese (0.436 mg/kg), but also selenium (0.164 mg/kg) and zinc (19.9 mg/kg). The study findings indicate that the use of sewage sludge in agriculture can cause secondary multi-trace element deficiencies in ruminants. Careful attention should be taken when crops are ensilaged avoiding top-soil sewage sludge contamination, since the acidification process may greatly increase Fe bioavailability.

Umbilical cord mesenchymal stromal cells (UC-MSC) are promising candidates for cell therapy due to their potent multilineage differentiation, enhanced self-renewal capacity, and immediate availability for clinical use. Clinical experience has demonstrated satisfactory biosafety profiles and feasibility of UC-MSC application in the allogeneic setting. However, the use of UC-MSC for bone regeneration has not been fully established. A major challenge in the generation of successful therapeutic strategies for bone engineering lies on the combination of highly functional proosteogenic MSC populations and bioactive matrix scaffolds. To address that, in this study we proposed a new approach for the generation of bone-like constructs based on UC-MSC expanded in human platelet lysate (hPL) and evaluated its potential to induce bone structures in vivo . In order to obtain UC-MSC for potential clinical use, we first assessed parameters such as the isolation method, growth supplementation, microbiological monitoring, and cryopreservation and performed full characterization of the cell product including phenotype, growth performance, tree-lineage differentiation, and gene expression. Finally, we evaluated bone-like constructs based on the combination of stimulated UC-MSC and collagen microbeads for in vivo bone formation. UC-MSC were successfully cultured from 100% of processed UC donors, and efficient cell derivation was observed at day 14 ± 3 by the explant method. UC-MSC maintained mesenchymal cell morphology, phenotype, high cell growth performance, and probed multipotent differentiation capacity. No striking variations between donors were recorded. As expected, UC-MSC showed tree-lineage differentiation and gene expression profiles similar to bone marrow- and adipose-derived MSC. Importantly, upon osteogenic and endothelial induction, UC-MSC displayed strong proangiogenic and bone formation features. The combination of hPL-expanded MSC and collagen microbeads led to bone/vessel formation following implantation into an immune competent mouse model. Collectively, we developed a high-performance UC-MSC-based cell manufacturing bioprocess that fulfills the requirements for human application and triggers the potency and effectivity of cell-engineered scaffolds for bone regeneration., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2019 Ingrid Silva-Cote et al.)