Search

Background: Barth syndrome (BTHS) is caused by mutations of the gene encoding tafazzin, which catalyzes maturation of mitochondrial cardiolipin and often manifests with systolic dysfunction during early infancy. Beyond the first months of life, BTHS cardiomyopathy typically transitions to a phenotype of diastolic dysfunction with preserved ejection fraction, blunted contractile reserve during exercise, and arrhythmic vulnerability. Previous studies traced BTHS cardiomyopathy to mitochondrial formation of reactive oxygen species (ROS). Because mitochondrial function and ROS formation are regulated by excitation-contraction coupling, integrated analysis of mechano-energetic coupling is required to delineate the pathomechanisms of BTHS cardiomyopathy. Methods: We analyzed cardiac function and structure in a mouse model with global knockdown of tafazzin ( Taz -KD) compared with wild-type littermates. Respiratory chain assembly and function, ROS emission, and Ca 2+ uptake were determined in isolated mitochondria. Excitation-contraction coupling was integrated with mitochondrial redox state, ROS, and Ca 2+ uptake in isolated, unloaded or preloaded cardiac myocytes, and cardiac hemodynamics analyzed in vivo. Results: Taz -KD mice develop heart failure with preserved ejection fraction (>50%) and age-dependent progression of diastolic dysfunction in the absence of fibrosis. Increased myofilament Ca 2+ affinity and slowed cross-bridge cycling caused diastolic dysfunction, in part, compensated by accelerated diastolic Ca 2+ decay through preactivated sarcoplasmic reticulum Ca 2 + -ATPase. Taz deficiency provoked heart-specific loss of mitochondrial Ca 2+ uniporter protein that prevented Ca 2+ -induced activation of the Krebs cycle during β-adrenergic stimulation, oxidizing pyridine nucleotides and triggering arrhythmias in cardiac myocytes. In vivo, Taz -KD mice displayed prolonged QRS duration as a substrate for arrhythmias, and a lack of inotropic response to β-adrenergic stimulation. Cellular arrhythmias and QRS prolongation, but not the defective inotropic reserve, were restored by inhibiting Ca 2+ export through the mitochondrial Na + /Ca 2+ exchanger. All alterations occurred in the absence of excess mitochondrial ROS in vitro or in vivo. Conclusions: Downregulation of mitochondrial Ca 2+ uniporter, increased myofilament Ca 2+ affinity, and preactivated sarcoplasmic reticulum Ca 2+ -ATPase provoke mechano-energetic uncoupling that explains diastolic dysfunction and the lack of inotropic reserve in BTHS cardiomyopathy. Furthermore, defective mitochondrial Ca 2+ uptake provides a trigger and a substrate for ventricular arrhythmias. These insights can guide the ongoing search for a cure of this orphaned disease.

Rationale: Renin-expressing cells are myoendocrine cells crucial for survival. They have been postulated to possess a pressure-sensing mechanism, a baroreceptor, which can detect slight changes in blood pressure and respond with precise and synchronized amounts of renin synthesized and released to the circulation to maintain blood pressure and fluid-electrolyte balance. The location and nature of this puzzling pressure-sensing structure have remained unknown as it was originally suggested over 60 years ago. Objective: To elucidate the location and structure of the renin cell baroreceptor. Methods and Results: We used a variety of genetically modified mice whereby renin cells were exposed in vivo to either low or high arterial pressure. In addition, we applied direct mechanical stimuli, that is, pneumatic pressure or stretch, directly to renin cells cultured under different conditions and substrata. Changes in perfusion pressure and/or direct mechanical stimuli induced significant changes in renin gene expression and the phenotype of renin cells. Importantly, the experiments show that the pressure-sensing mechanism (the baroreceptor) resides in the renin cells; it requires initial extracellular sensing by integrin β1 at the renin cell membrane and is transduced to the nuclear membrane and chromatin by lamin A/C. Conclusions: These studies show that the enigmatic baroreceptor is a nuclear mechanotransducer that resides in the renin cells per se and is responsible for the sensing and transmission of extracellular physical forces directly to the chromatin of renin cells via lamin A/C to regulate renin gene expression, renin bioavailability, and homeostasis.

Organochlorine pesticides have generated public concern worldwide because of their toxicity to human health and the environment, even at low concentrations, and their persistence, being mostly nonbiodegradable. The use of 2,4-dichlorophenoxyacetic acid (2,4-D) has increased in recent decades, causing severe water contamination. Several treatments have been developed to degrade 2,4-D. This manuscript presents an overview of the physicochemical characteristics, uses, regulations, environmental and human health impacts of 2,4-D, and different advanced oxidation processes (AOPs) to degrade this organic compound, evaluating and comparing operation conditions, efficiencies, and intermediaries. Based on this review, 2,4-D degradation is highly efficient in ozonation (system O3/plasma, 99.8% in 30 min). Photocatalytic, photo-Fenton, and electrochemical processes have the optimal efficiencies of degradation and mineralization: 97%/79.67% (blue TiO2 nanotube arrays//UV), 100%/98% (Fe2+/H2O2/UV), and 100%/84.3% (MI-meso SnO2), respectively. The ozonation and electrochemical processes show high degradation efficiencies, but energy costs are also high, and photocatalysis is more expensive with a separation treatment used to recover the catalyst in the solution. The Fenton process is a viable economic-environmental option, but degradation efficiencies are often low (50–70%); however, they are increased when solar UV radiation is used (90–100%). AOPs are promising technologies for the degradation of organic pollutants in real wastewater, so evaluating their strengths and weaknesses is expected to help select viable operational conditions and obtain optimal efficiencies.

Rooting induction of leafy stem cuttings of Hevea brasiliensis can be used as a relevant tool for the cultivation, conservation of genetic diversity, and rescue of this species in Peru, decimated by decades of extractive practices. In this study, we evaluated the effects of different numbers of leaflets and concentrations of indole-3-butyric acid (IBA) in the rooting capacity of leafy stem cuttings of H. brasiliensis. Cuttings 6–7 cm long were collected from stockplants in a germplasm bank; leaves were pruned to 2 or 5 leaflets, and the cutting base was treated with one of the following IBA concentrations: 0, 1000, 2000, 3000, 4000, or 5000 ppm. Cuttings were placed in trays with a sandy substrate and transferred to plastic mini-tunnels with fogging irrigation. After 29 days, cuttings with 5 leaflets and 2000 ppm of IBA showed the best results in survival (100%), rooting percentage (79%), and roots number (4.1) and length (2.9 cm). The high conversion rate of cuttings to plants indicates the feasibility of this technique as a simple and effective tool, both for the conservation and the commercial propagation of selected rubber tree genotypes.

The physicochemical treatment (PT) of food industry wastewater was investigated. In the first stage, calcium magnesium acetate (CaMgAc4) was synthesized using eggshell (biocalcium), magnesium oxide...

Testicular germ cell tumors (TGCTs) are the most common cancers in men aged 15–39 years and are divided into two major groups, seminomas and nonseminomas. Novel treatment options are required for these patients, to limit side effects of chemotherapy. We hypothesized that promoter methylation of relevant homologous recombination (HR) genes might be predictive of response to poly‐ADP ribose polymerase inhibitors (PARPis) in TGCTs. We report a study pipeline combining in silico, in vitro, and clinical steps. By using several databases and in silico tools, we identified BRCA1, RAD51C, PALB2, RAD54B, and SYCP3 as the most relevant genes for further investigation and pinpointed specific CpG sites with pronounced negative correlation to gene expression. Nonseminomas displayed significantly higher methylation levels for all target genes, where increased methylation was observed in patients with more differentiated subtypes and higher disease burden. We independently performed second‐line targeted validation in tissue series from TGCT patients. A moderate and/or strong anti‐correlation between gene expression (assessed by RNA‐sequencing) and promoter methylation (assessed by 450k array) was found, for all of the targets. As a proof of concept, we demonstrated the sensitivity of TGCT cell lines to Olaparib, which associated with differential methylation levels of a subset of targets, namely BRCA1 and RAD51C. Our findings support the use of HR genes promoter methylation as a predictor of the therapeutic response to PARPis in patients with TGCT., We report a study pipeline for exploring whether aberrant promoter methylation of genes involved in the homologous recombination pathway determines responsiveness to PARP inhibitors in testicular germ cell tumor patients. In silico analyses were followed by validation using quantitative methylation‐specific PCR. We showed differential response of germ cell tumor cell lines to Olaparib, associated with BRCA1/RAD51C promoter methylation.

To compare the regenerative properties of human stem cells of the apical papilla (SCAPs) embedded in a platelet-rich plasma (PRP) scaffold, when implanted in vivo using an organotypic model composed of human root segments, with or without the presence of the bioactive cements – ProRoot MTA or Biodentine. SCAPs were isolated from third molars with incomplete rhizogenesis and expanded and characterized in vitro using stem cell and surface markers. The pluripotency of these cells was also assessed using adipogenic, chondrogenic, and osteogenic differentiation protocols. SCAPs together with a scaffold of PRP were added to the root segment lumen and the organotypic model implanted on the dorsal region of immunodeficient rats for a period of 4 months. Presence of SCAPs induced de novo formation of dentin-like and pulp-like tissue. A barrier of either ProRoot MTA or Biodentine did not significantly affect the fraction of sections from roots segments observed to contain deposition of hard material (P > 0.05). However, the area of newly deposited dentin was significantly greater in segments containing a barrier of Biodentine compared with ProRoot MTA (P < 0.001). SCAPs offer a viable alternative to other dental stem cells (DSCs) in their regenerative properties when enclosed in the microenvironment of human tooth roots. The present study also suggests that the presence of bioactive materials does not hinder or impede the formation of new hard tissues, but the presence of Biodentine may promote greater mineralized tissue deposition.

Bacterial spores of the genus Bacillus are being evaluated as adjuvant molecules capable of improving the immune response to vaccines. In this study, we investigate whether subcutaneously administered spores of B. toyonensis BCT-7112T could enhance a vaccine immune response in mice. Three groups of mice were subcutaneously vaccinated on day 0 and received a booster on day 21 of the experiment, with the following vaccine formulations: 40 µg of recombinant glycoprotein D (rgD) from bovine herpesvirus type 5 (BoHV-5) adsorbed in 10% aluminum hydroxide (alum) without B. toyonensis spores (group 1) and B. toyonensis (1 × 106 viable spores) + 40 µg of rgD adsorbed in 10% alum (group 2); and B. toyonensis (1 × 106 viable spores) without rgD (group 3). Group 2 showed significantly higher titers (P

Bacterial nonulosonic acids such as pseudaminic acids and others constitute a family of 9-carbon monosaccharides that contain a common 3-deoxy-2-ketoacid fragment but differ in their stereochemistries at 5 stereogenic centers between C-4 to C-8. Their unique structures make them attractive targets for use as antigens in vaccinations to combat drug-resistant bacterial infections and their challenging stereochemistries have attracted considerable attention from chemists. In this work we report the development of an improved synthesis for 2,4-di-N-acetyl-L-altrose (L-2,4-Alt-diNAc), which is a key hexose required for the chemical and chemoenzymatic synthesis of pseudaminic acids. Using L-fucose as a starting material, our synthesis overcomes several pitfalls in previously reported syntheses.

We demonstrate the fabrication of carbon nanoribbons with a width of 40 nm based on fixation and pyrolysis of an organic template, lipid nanotubes. To our best knowledge, this is the smallest feature size achieved by pyrolysis of surface-patterned organic templates. Such a pyrolytic carbon nanostructure can be used for electronics and sensing applications in future.

Purpose: Carcinoma cervix forms a major burden among malignancies in women in India. Anemia is very common among Indian women and a high proportion of carcinoma cervix patients present with anemia. Previous studies show that hemoglobin level may be considered as a prognostic factor in carcinoma cervix. It has been observed that anemia is associated with diminished radiocurability in patients with carcinoma of the cervix. However, the therapeutic benefit of the measures to correct anemia remains debatable. The objective of this study was to evaluate the association of hemoglobin concentrations and its correction by blood transfusions before and during treatment and the response to chemoradiation. Materials and Methods: Fifty patients with histologically proven squamous cell carcinoma of the cervix (Stage II to IV) from December 2018 to May 2020 were taken up for this study. Patients received definitive chemoradiation of 50 Gy in 25 fractions, 2 Gy/fraction, 5 fractions per week, along with-weekly injection cisplatin/carboplatin, followed by brachytherapy. All patients had their pretreatment hemoglobin levels measured at the first consultation. The effects of hemoglobin concentrations at presentation and during therapy on the clinical response to radiotherapy were studied. Blood transfusions and the time point when transfusions were given were recorded. Results: Eighteen patients received packed red blood cell transfusion before the start of radiation therapy and 9 patients received packed cell transfusion during radiation therapy. Out of the 18 patients who received transfusion before RT, 3 patients had complete response. Out of the 9 patients who received transfusion during radiation, 4 had complete response. The presence of anemia had a significant impact on the response rate. Pretreatment anemia was associated with poor response and local control among patients who underwent chemoradiotherapy for cervical cancer. Conclusion: Pretreatment hemoglobin and hemoglobin levels during radiation therapy were prognostic factors for local control, in addition to stage and histology, which might have independently predicted the outcome.

BackgroundMalaria caused byPlasmodium falciparumin pregnancy can result in adverse maternal and fetal sequelae. This review evaluated the adherence of the national guidelines drawn from World Health Organization (WHO) regions, Africa, Eastern Mediterranean, Southeast Asia, and Western Pacific, to the WHO recommendations on drug treatment and prevention of chloroquine-resistant falciparum malaria in pregnant women.MethodsThirty-five updated national guidelines and the President’s Malaria Initiative (PMI), available in English language, were reviewed. The primary outcome measures were the first-line anti-malarial treatment protocols adopted by national guidelines for uncomplicated and complicated falciparum malaria infections in early (first) and late (second and third) trimesters of pregnancy. The strategy of intermittent preventive treatment of malaria in pregnancy (IPTp) with sulfadoxine-pyrimethamine (SP) was also addressed.ResultsThis review evaluated the treatment and prevention of falciparum malaria in pregnancy in 35 national guidelines/PMI-Malaria Operational Plans (MOP) reports out of 95 malaria-endemic countries. Of the 35 national guidelines, 10 (28.6%) recommend oral quinine plus clindamycin as first-line treatment for uncomplicated malaria in the first trimester. As the first-line option, artemether–lumefantrine, an artemisinin-based combination therapy, is adopted by 26 (74.3%) of the guidelines for treating uncomplicated or complicated malaria in the second and third trimesters. Intravenous artesunate is approved by 18 (51.4%) and 31 (88.6%) guidelines for treating complicated malaria during early and late pregnancy, respectively. Of the 23 national guidelines that recommend IPTp-SP strategy, 8 (34.8%) are not explicit about directly observed therapy requirements, and three-quarters, 17 (73.9%), do not specify contra-indication of SP in human immunodeficiency virus (HIV)-infected pregnant women receiving cotrimoxazole prophylaxis. Most of the guidelines (18/23; 78.3%) state the recommended folic acid dose.ConclusionSeveral national guidelines and PMI reports require update revisions to harmonize with international guidelines and emergent trends in managing falciparum malaria in pregnancy. National guidelines and those of donor agencies should comply with those of WHO guideline recommendations although local conditions and delayed guideline updates may call for deviations from WHO evidence-based guidelines.

Diabetes mellitus and its complication such as heart disease, stroke, kidney failure, etc. is a serious concern all over the world. Hence, monitoring some important blood parameters non-invasively is of utmost importance, that too with high accuracy. This paper presents an in-house developed system, which will be helpful for diabetes patients with Chronic Kidney Disease (CKD) to monitor blood urea and glucose. This manuscript discusses a comparative study for the prediction of blood urea and glucose using Backpropagation Artificial Neural Network (BP- ANN) and Partial Least Square Regression (PLSR) model. The NVIDIA Jetson Nano board controls the five fixed LED wavelengths in the Near Infrared (NIR) region from \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} }{}$2.0~\mu \text{m}$\end{document} to \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{upgreek} \usepackage{mathrsfs} \setlength{\oddsidemargin}{-69pt} \begin{document} }{}$2.5~\mu \text{m}$ \end{document} with a constant emission power of 1.2 mW. The spectra for 57 laboratory prepared samples conforming with major blood constituents of the blood sample were recorded. From these samples, 53 spectra were used for training/calibration of the BP-ANN/PLSR model and the remaining 4 samples were used for validating the model. The PLSR model predicts blood urea and glucose with a Root Mean Square Error (RMSE) of 0.88 & 12.01 mg/dL, Coefficient of Determination R2 = 0.93 & R2 = 0.97, Accuracy of 94.2 % and 90.14 %, respectively. To improve the prediction accuracy, BP-ANN model is applied. Later the Principal Component Analysis (PCA) technique was applied to these 57 spectra values. These PCA values were used to train and validate the BP-ANN model. After applying the BP-ANN model, the prediction of blood urea & glucose improved remarkably, which achieved RMSE of 0.69 mg/dL, R2 = 0.96, Accuracy of 95.96 % for urea and RMSE of 2.06 mg/dL, R2 = 0.99, and Accuracy of 98.65 % for glucose. The system performance is then evaluated with Bland Altman analysis and Clarke Error Grid Analysis (CEGA).

Large-scale production of graphene sheets by liquid-phase exfoliation of graphite is a challenging task from a sustainability point of view. Certain bio-derived solvents were found to exfoliate graphite to produce single-layered graphene sheets but the high cost of the solvent is always a deterring factor towards upscaling of the process. Herein, Kappaphycus alvarezii, a cultivable red seaweed is demonstrated as a sustainable resource for producing a bio solvent for exfoliation and to produce graphene sheets from graphite. A solvent system consisting of levulinic acid, acetic acid, and γ-valerolactone was prepared from the polysaccharide obtained from the seaweed biomass through acid hydrolysis under pressure and the mixture was found to exfoliate graphite to produce few-layered pristine graphene nanosheets. The process is scalable and cost-effective and the seaweed biomass-derived solvent mixture can be recovered and reused in the subsequent cycles of exfoliation for large-scale production of graphene nanosheets.

The oxidation of tyrosine to form the neutral tyrosine radical via proton-coupled electron transfer is essential for a wide range of biological processes. The precise measurement of the proton-coupled redox potentials of tyrosine (Y) in complex protein environments is challenging mainly due to the highly oxidizing and reactive nature of the radical state. Herein a computational strategy is presented for predicting proton-coupled redox potentials in a protein environment. In this strategy, both the reduced Y-OH and oxidized Y-O(•) forms of tyrosine are sampled with molecular dynamics using a molecular mechanical force field. For a large number of conformations, a quantum mechanical/molecular mechanical (QM/MM) electrostatic embedding scheme is used to compute the free energy differences between the reduced and oxidized forms, including the zero-point energy and entropic contributions as well as the impact of the protein electrostatic environment. This strategy is applied to a series of fluorinated tyrosine derivatives embedded in a de novo α-helical protein denoted α(3)Y. The force fields for both the reduced and oxidized forms of these non-canonical fluorinated tyrosine residues are parameterized for general use. The calculated relative proton-coupled redox potentials agree with experimentally measured values with a mean unsigned error of 34 mV. Analysis of the simulations illustrate that hydrogen-bonding interactions between tyrosine and water increase the redox potentials by ~100–250 mV, with significant variations due to the fluctuating protein environment. This QM/MM approach enables the calculation of proton-coupled redox potentials of tyrosine and other residues such as tryptophan in a variety of protein systems.

Nonviral vectors are gaining importance as gene delivery systems. However, to meet the large-scale applications, their sustainable production is of utmost importance. Hence, a cost-effective and on...

In this work, binary MPt (M = Co, Ni, Cu) nanoalloys and mesoporous carbon nitride (mpg-C3N4) as support material are prepared, and then gathered together by liquid phase self-assembly in order to produce MPt/mpg-C3N4 electrocatalysts. The yielded electrocatalysts are characterized by TEM, ICP-MS and XRD techniques in order to determine their morphology, composition and textural properties. Evaluation of MPt/mpg-C3N4 (

In this study, acetaminophen (ACT) in aqueous solution was treated with electrooxidation and photo-electrooxidation processes (PEO). An electrochemical cell was used for the treatment of different concentrations of ACT (10, 50 and 80 mg L

OBJECTIVE: It has been reported that glutathione (GSH), the most abundant cellular antioxidant, can inhibit production of pro-inflammatory cytokines by activated macrophages. Bromosulfophthalein (BSP) has been recognized as an inhibitor of the efflux of reduced GSH from cells, leading to an increase in the intracellular GSH level. In this study, we evaluated, for the first time, whether BSP possessed anti-inflammatory effects in lipopolysaccharide (LPS)-stimulated macrophages. MATERIALS AND METHODS: RAW 264.7 cells were treated with BSP and the levels of proinflammatory cytokines, GSH, and nitrite were assessed. Gene expression of inducible nitric oxide synthase (iNOS), tumor necrosis factor alpha (TNF α), interleukin-1β (IL-1β) and interleukin-6 (IL-6) was analyzed via quantitative RT-PCR. We also examined various inflammatory signaling pathways including Akt/forkhead box protein O1 (FoxO1)/ toll-like receptor 4 (TLR4), mitogen-activated protein kinases (MAPKs) and Fas proteins by Western blot and flow cytometry analysis. RESULTS: Our study demonstrated that BSP induced an increase in intracellular GSH levels in LPS-stimulated macrophages. BSP inhibited production of nitric oxide and proinflammatory cytokines. BSP increased phosphorylation of Akt and nuclear exclusion of FoxO1 and suppressed TLR4 expression. Additionally, BSP decreased MAPKs activation and Fas expression. DISCUSSION AND CONCLUSIONS: Taken together, these data suggest that BSP can attenuate inflammation through multiple signaling pathways. These findings highlightthe potential of BSP as a new anti-inflammatory agent.

Intra-articular (IA) injection is an attractive route of administration for the treatment of osteoarthritis (OA). However, free drugs injected into the joint space are rapidly cleared and many of them can induce adverse off-target effects on different IA tissues. To overcome these limitations, we designed nanocomposite 4-arm-poly(ethylene glycol)-maleimide (PEG-4MAL) microgels, presenting cartilage- or synoviocyte-binding peptides, containing poly(lactic-co-glycolic) acid (PLGA) nanoparticles (NPs) as an IA small molecule drug delivery system. Microgels containing rhodamine B (model drug)-loaded PLGA NPs were synthesized using microfluidics technology and exhibited a sustained, near zero-order release of the fluorophore over 16 days in vitro. PEG-4MAL microgels presenting synoviocyte- or cartilage-targeting peptides specifically bound to rabbit and human synoviocytes or to bovine articular cartilage in vitro, respectively. Finally, using a rat model of post-traumatic knee OA, PEG-4MAL microgels were shown to be retained in the joint space for at least 3 weeks without inducing any joint degenerative changes as measured by EPIC-μCT and histology. Additionally, all microgel formulations were found trapped in the synovial membrane and significantly increased the IA retention time of a model small molecule near-infrared (NIR) dye compared to that of the free dye. These results suggest that peptide-functionalized nanocomposite PEG-4MAL microgels represent a promising intra-articular vehicle for tissue-localized drug delivery and prolonged IA drug retention for the treatment of OA.

Polyhydroxyalkanoates (PHAs) have a wide range of applicability due to their biodegradable and biocompatible characteristics. This makes them suitable for their commercialization as an alternative ...

Esta investigación respecto a la calidad y manejo de los suelos agrícolas fue realizada en 18 sistemas de producción agrícolas de la colonia Providencia, seleccionados a través de un muestreo estratificado, estos se organizaron en 3 grupos según el tamaño de las áreas agrícolas, se evaluaron características físicas, químicas y biológicas del suelo, así como las actividades de manejo que realizan los productores. Los análisis físico-químicos se realizaron en el laboratorio de suelos y agua de la Universidad Nacional Agraria.En cuanto al uso actual de los suelos agrícolas predominan los granos básicos, raíces y tubérculos y café robusta, son suelos manejados de forma tradicional con indicios de degradación física (hídrica y eólica) y biológica, medianamente ácidos, con materia orgánica y nitrógeno adecuados para satisfacer la demanda de los cultivos, el fósforo es deficiente y el potasio moderadamente disponible.Físicamente son suelos ultisoles, poco profundos con dos horizontes definidos, color rojizo oscuro, arcilloso, generalmente en estructura de bloques y angulares, muchos poros finos y poco finos, plástico adherente con contenido de materia orgánica superior al 5%, medianamente ácidos

Renin cells are crucial for the regulation of blood pressure and fluid electrolyte homeostasis. We have recently shown that renin cells possess unique chromatin features at regulatory regions throughout the genome that may determine the identity and memory of the renin phenotype. The 3-D structure of chromatin may be equally important in the determination of cell identity and fate. CCCTC-binding factor (Ctcf) is a highly conserved chromatin organizer that may regulate the renin phenotype by controlling chromatin structure. We found that Ctcf binds at several conserved DNA sites surrounding and within the renin locus, suggesting that Ctcf may regulate the transcriptional activity of renin cells. In fact, deletion of Ctcf in cells of the renin lineage led to decreased endowment of renin-expressing cells accompanied by decreased circulating renin, hypotension, and severe morphological abnormalities of the kidney, including defects in arteriolar branching, and ultimately renal failure. We conclude that control of chromatin architecture by Ctcf is necessary for the appropriate expression of renin, control of renin cell number and structural integrity of the kidney.

Key points The Anrep effect represents the alteration of left ventricular (LV) contractility to acutely enhanced afterload in a few seconds, thereby preserving stroke volume (SV) at constant preload. As a result of the missing preload stretch in our model, the Anrep effect differs from the slow force response and has a different mechanism. The Anrep effect demonstrated two different phases. First, the sudden increased afterload was momentary equilibrated by the enhanced LV contractility as a result of higher power strokes of strongly-bound myosin cross-bridges. Second, the slightly delayed recovery of SV is perhaps dependent on Ca2+ /calmodulin-dependent protein kinase II activation caused by oxidation and myofilament phosphorylation (cardiac myosin-binding protein-C, myosin light chain 2), maximizing the recruitment of available strongly-bound myosin cross-bridges. Short-lived oxidative stress might present a new facet of subcellular signalling with respect to cardiovascular regulation. Relevance for human physiology was demonstrated by echocardiography disclosing the Anrep effect in humans during handgrip exercise. Abstract The present study investigated whether oxidative stress and Ca2+ /calmodulin-dependent protein kinase II (CaMKII) activity are involved in triggering the Anrep effect. LV pressure-volume (PV) analyses of isolated, preload controlled working hearts were performed at two afterload levels (60 and 100 mmHg) in C57BL/6N wild-type (WT) and CaMKII-double knockout mice (DKOCaMKII ). In snap-frozen WT hearts, force-pCa relationship, H2 O2 generation, CaMKII oxidation and phosphorylation of myofilament and Ca2+ handling proteins were assessed. Acutely raised afterload showed significantly increased wall stress, H2 O2 generation and LV contractility in the PV diagram with an initial decrease and recovery of stroke volume, whereas end-diastolic pressure and volume, as well as heart rate, remained constant. Afterload induced increase in LV contractility was blunted in DKOCaMKII -hearts. Force development of single WT cardiomyocytes was greater with elevated afterload at submaximal Ca2+ concentration and associated with increases in CaMKII oxidation and phosphorylation of cardiac-myosin binding protein-C, myosin light chain and Ca2+ handling proteins. CaMKII activity is involved in the regulation of the Anrep effect and associates with stimulation of oxidative stress, presumably starting a cascade of CaMKII oxidation with downstream phosphorylation of myofilament and Ca2+ handling proteins. These mechanisms improve LV inotropy and preserve stroke volume within few seconds.

Key points Mitochondrial Ca2+ uptake stimulates the Krebs cycle to regenerate the reduced forms of pyridine nucleotides (NADH, NADPH and FADH2 ) required for ATP production and reactive oxygen species (ROS) elimination. Ca2+ /calmodulin-dependent protein kinase II (CaMKII) has been proposed to regulate mitochondrial Ca2+ uptake via mitochondrial Ca2+ uniporter phosphorylation. We used two mouse models with either global deletion of CaMKIIδ (CaMKIIδ knockout) or cardiomyocyte-specific deletion of CaMKIIδ and γ (CaMKIIδ/γ double knockout) to interrogate whether CaMKII controls mitochondrial Ca2+ uptake in isolated mitochondria and during β-adrenergic stimulation in cardiac myocytes. CaMKIIδ/γ did not control Ca2+ uptake, respiration or ROS emission in isolated cardiac mitochondria, nor in isolated cardiac myocytes, during β-adrenergic stimulation and pacing. The results of the present study do not support a relevant role of CaMKII for mitochondrial Ca2+ uptake in cardiac myocytes under physiological conditions. Abstract Mitochondria are the main source of ATP and reactive oxygen species (ROS) in cardiac myocytes. Furthermore, activation of the mitochondrial permeability transition pore (mPTP) induces programmed cell death. These processes are essentially controlled by Ca2+ , which is taken up into mitochondria via the mitochondrial Ca2+ uniporter (MCU). It was recently proposed that Ca2+ /calmodulin-dependent protein kinase II (CaMKII) regulates Ca2+ uptake by interacting with the MCU, thereby affecting mPTP activation and programmed cell death. In the present study, we investigated the role of CaMKII under physiological conditions in which mitochondrial Ca2+ uptake matches energy supply to the demand of cardiac myocytes. Accordingly, we measured mitochondrial Ca2+ uptake in isolated mitochondria and cardiac myocytes harvested from cardiomyocyte-specific CaMKII δ and γ double knockout (KO) (CaMKIIδ/γ DKO) and global CaMKIIδ KO mice. To simulate a physiological workload increase, cardiac myocytes were subjected to β-adrenergic stimulation (by isoproterenol superfusion) and an increase in stimulation frequency (from 0.5 to 5 Hz). No differences in mitochondrial Ca2+ accumulation were detected in isolated mitochondria or cardiac myocytes from both CaMKII KO models compared to wild-type littermates. Mitochondrial redox state and ROS production were unchanged in CaMKIIδ/γ DKO, whereas we observed a mild oxidation of mitochondrial redox state and an increase in H2 O2 emission from CaMKIIδ KO cardiac myocytes exposed to an increase in workload. In conclusion, the results obtained in the present study do not support the regulation of mitochondrial Ca2+ uptake via the MCU or mPTP activation by CaMKII in cardiac myocytes under physiological conditions.

Four different ionic liquids were synthesized, characterized and used with polypropylene glycol for the formation of aqueous biphasic systems and were used to selectively separate e-PL from culture...

The efficacy of commercially available chemical insecticides and biopesticides on the cotton mealybug (CMB), Phenacoccus solenopsis, was evaluated in the glasshouse. Spirotetramat, sulfoxaflor and buprofezin were identified as key insecticides for use in integrated pest management (IPM) strategies aimed at controlling CMB without flaring other co-occurring pests. When used as a single application, spirotetramat and sulfoxaflor at the rate of 96 g (active ingredient, ha−1) provided variable control of CMB. Spirotetramat used in a double spray tactic (two sequential sprays, 14–15 days apart) without crop oil provided ≥80% control of adult CMB while the addition of oil (5% v/v) increased control to ≥90%. Clothianidin synergised the spirotetramat + oil combination and was identified as a potentially useful tank mix option for use in situations where a quick knockdown of high density and/or large infestation of CMB is required, or to treat high risk infestations in squaring or younger cotton when the abundance of beneficial insects is typically low. Sulfoxaflor used in a double spray tactic provided ≥90% control of adult CMB. The addition of Pulse® penetrant (0.5% v/v) to both options improved overall efficacy. Addition of crop oil to sulfoxaflor did not yield any tangible benefits. Spirotetramat and buprofezin were identified as important tools in managing situations where whitefly (Bemisia tabaci) is the primary pest management target, but CMB is also present in the crop. Buprofezin was effective on early instar mealybugs; this makes it an option for arresting CMB population growth while allowing the beneficial insect populations to increase. Sulfoxaflor was shown to be a useful option in situations where CMB is present along with key pests such as mirids (Creontiades spp.). Mealybugs are typically well controlled by naturally occurring beneficial insects without the need for insecticide use. Chemical insecticides for CMB control should be considered only as a last resort and deployed within the bounds of an IPM strategy.

Background and aims Dietary long-chain (≥20 carbons) n-3 polyunsaturated fatty acids (PUFAs) reduce atherosclerosis and enhance macrophage autophagy activation. How macrophage autophagy impacts atherosclerotic progression, particularly when comparing dietary n-3 PUFA supplementation vs. saturated fat feeding, is unknown. Methods We generated myeloid-specific autophagy-deficient and control mice in the Ldlr−/− background by transplanting bone marrow from myeloid-specific autophagy-related (atg) 5 knockout mice and wild type controls into irradiated Ldlr−/− recipients. After 7 weeks for recovery from radiation, mice were fed an atherogenic diet containing 0.2% cholesterol and 20% calories as palm oil (PO diet), or 10% calories as PO plus 10% calories as fish oil (FO diet) for 16 weeks. Results Compared to PO, FO significantly reduced plasma cholesterol, triglyceride, hepatic neutral lipid, and aortic caspase-1 cleavage, but increased aortic efferocytosis, leading to attenuated atherosclerosis in Ldlr−/− mice receiving wild type bone marrow. Myeloid atg5 deletion had little impact on plasma lipid concentrations and hepatic neutral lipid content, regardless of diet. Myeloid atg5 deletion increased aortic caspase-1 cleavage, decreased aortic efferocytosis and worsened atherosclerosis only in the FO-fed Ldlr−/− mice. Conclusions Deficient myeloid autophagy significantly attenuated FO-induced atheroprotection, suggesting that dietary n-3 PUFAs reduce atherosclerosis, in part, by activation of macrophage autophagy.

This study aimed to estimate chronic daily intake (CDI) and to predict the attributable lifetime cancer risk (LCR) and hazard index (HI) from concurrent exposure to four trihalomethanes (THMs; chloroform, bromodichloromethane, dibromochloromethane and bromoform), via multiple exposure routes (oral ingestion, dermal contact and inhalation), among 238 non-competitive attendees of 10 Portuguese public indoor swimming pools (SPs), using a probabilistic approach based on Monte Carlo simulations. Exposure parameters of study participants were collected via questionnaires and THMs levels in SPs water were determined according the respective normative standards. The CDI for total THMs calculated for male and female participants considering all routes was 7.52 and 8.97 mg/kg/day, respectively. SP attendees presented higher CDI through inhalation than via the other two exposure routes, and chloroform was the compound contributing the most to total THMs CDI. The risk analysis indicated that the total LCR and HI from the targeted THMs were higher than the negligible risk levels (1 × 10-6 and 1, respectively) in the scenarios examined (central tendency exposure and reasonable maximum exposure), and the health risk for females was slightly higher than for males. This study suggests that there are possible adverse health risks, thus, to protect pool attendees, adequate mitigation measures, and comprehensive regulatory guidelines on individual THMs concentrations are needed. Highlights: Chloroform was the main THM; inhalation route was found to be the most relevant; The carcinogenic risk exceeds U.S. EPA negligible level; The non-carcinogenic risk is several orders greater than the recommended value; Females are more susceptible to cancer and non-cancer risk from total THMs exposure. This work was supported by the Project NORTE-01-0145-FEDER 000010, Health, Comfort and Energy in the Built Environment (HEBE), co-financed by Programa Operacional Regional do Norte (NORTE2020), through the European Regional Development Fund and by the Foundation for Science and Technology – FCT (Portuguese Ministry of Science, Technology and Higher Education) under the project UIDB/04750/2020. Joana Madureira and Carla Costa are supported by FCT (SFRH/BPD/115112/2016 and SFRH/BPD/96196/2013 grants, respectively). Ri cardo Assunção was supported by FCT Individual CEEC 2018 Assistant Researcher Grant CEECIND/01570/2018. The authors are also grateful for the support from Xunta de Galicia [ED431B 2019/02], Ministerio de Educación, Cultura y Deporte [BEAGAL18/00142 to V.V.], Programa IACOBUS Estancias de Investigación 2021; and FCT/MCTES for the financial support to CESAM (UIDP/50017/2020+UIDB/50017/2020) through national funds. info:eu-repo/semantics/publishedVersion