Search

COVID-19, caused by SARS-CoV-2, is a respiratory disease associated with inflammation and endotheliitis. Mechanisms underling inflammatory processes are unclear, but angiotensin converting enzyme 2 (ACE2), the receptor which binds the spike protein of SARS-CoV-2 may be important. Here we investigated whether spike protein binding to ACE2 induces inflammation in endothelial cells and determined the role of ACE2 in this process. Human endothelial cells were exposed to SARS-CoV-2 spike protein, S1 subunit (rS1p) and pro-inflammatory signaling and inflammatory mediators assessed. ACE2 was modulated pharmacologically and by siRNA. Endothelial cells were also exposed to SARS-CoV-2. rSP1 increased production of IL-6, MCP-1, ICAM-1 and PAI-1, and induced NFkB activation via ACE2 in endothelial cells. rS1p increased microparticle formation, a functional marker of endothelial injury. ACE2 interacting proteins involved in inflammation and RNA biology were identified in rS1p-treated cells. Neither ACE2 expression nor ACE2 enzymatic function were affected by rSP1. Endothelial cells exposed to SARS-CoV-2 virus did not exhibit viral replication. We demonstrate that rSP1 induces endothelial inflammation via ACE2 through processes that are independent of ACE2 enzymatic activity and viral replication. We define a novel role for ACE2 in COVID-19- associated endotheliitis. [ABSTRACT FROM AUTHOR]

COVID-19, caused by SARS-CoV-2, is a respiratory disease associated with inflammation and endotheliitis. Mechanisms underling inflammatory processes are unclear, but angiotensin converting enzyme 2 (ACE2), the receptor which binds the spike protein of SARS-CoV-2 may be important. Here we investigated whether spike protein binding to ACE2 induces inflammation in endothelial cells and determined the role of ACE2 in this process. Human endothelial cells were exposed to SARS-CoV-2 spike protein, S1 subunit (rS1p) and pro-inflammatory signaling and inflammatory mediators assessed. ACE2 was modulated pharmacologically and by siRNA. Endothelial cells were also exposed to SARS-CoV-2. rSP1 increased production of IL-6, MCP-1, ICAM-1 and PAI-1, and induced NFkB activation via ACE2 in endothelial cells. rS1p increased microparticle formation, a functional marker of endothelial injury. ACE2 interacting proteins involved in inflammation and RNA biology were identified in rS1p-treated cells. Neither ACE2 expression nor ACE2 enzymatic function were affected by rSP1. Endothelial cells exposed to SARS-CoV-2 virus did not exhibit viral replication. We demonstrate that rSP1 induces endothelial inflammation via ACE2 through processes that are independent of ACE2 enzymatic activity and viral replication. We define a novel role for ACE2 in COVID-19- associated endotheliitis. [ABSTRACT FROM AUTHOR]

Aims NOX-derived reactive oxygen species (ROS) are mediators of signalling pathways implicated in vascular smooth muscle cell (VSMC) dysfunction in hypertension. Among the numerous redox-sensitive kinases important in VSMC regulation is c-Src. However, mechanisms linking NOX/ROS to c-Src are unclear, especially in the context of oxidative stress in hypertension. Here, we investigated the role of NOX-induced oxidative stress in VSMCs in human hypertension focusing on NOX5, and explored c-Src, as a putative intermediate connecting NOX5-ROS to downstream effector targets underlying VSMC dysfunction. Methods and results VSMC from arteries from normotensive (NT) and hypertensive (HT) subjects were studied. NOX1,2,4,5 expression, ROS generation, oxidation/phosphorylation of signalling molecules, and actin polymerization and migration were assessed in the absence and presence of NOX5 (melittin) and Src (PP2) inhibitors. NOX5 and p22phox-dependent NOXs (NOX1–4) were down-regulated using NOX5 siRNA and p22phox-siRNA approaches. As proof of concept in intact vessels, vascular function was assessed by myography in transgenic mice expressing human NOX5 in a VSMC-specific manner. In HT VSMCs, NOX5 was up-regulated, with associated oxidative stress, hyperoxidation (c-Src, peroxiredoxin, DJ-1), and hyperphosphorylation (c-Src, PKC, ERK1/2, MLC20) of signalling molecules. NOX5 siRNA reduced ROS generation in NT and HT subjects. NOX5 siRNA, but not p22phox-siRNA, blunted c-Src phosphorylation in HT VSMCs. NOX5 siRNA reduced phosphorylation of MLC20 and FAK in NT and HT. In p22phox- silenced HT VSMCs, Ang II-induced phosphorylation of MLC20 was increased, effects blocked by melittin and PP2. NOX5 and c-Src inhibition attenuated actin polymerization and migration in HT VSMCs. In NOX5 transgenic mice, vascular hypercontractilty was decreased by melittin and PP2. Conclusion We define NOX5/ROS/c-Src as a novel feedforward signalling network in human VSMCs. Amplification of this system in hypertension contributes to VSMC dysfunction. Dampening the NOX5/ROS/c-Src pathway may ameliorate hypertension-associated vascular injury. [ABSTRACT FROM AUTHOR]

This paper consists of two first-person reports of linguists working in the documentation of endangered languages. We narrate two opposite situations: that of Guató, a language with only two speakers, survivors of a long process of colonization, and that of the language spoken by the Xinane people, a monolingual recently-contacted group. We make use of these two examples to problematize the concept of «isolation»: the Xinane situation is, in fact, one of de-isolation (contact), carefully planned and executed by young people, after three generations of isolation motivated by white violence. Vicente, on the other hand, one of the two last speakers of the guató language can be more literally characterized as one of voluntary isolation, despite the 400 years of contact of his fragmented people. We also discuss the challenges of creating fieldwork methodologies in these two extremely contrastive contexts of language vitality. [ABSTRACT FROM AUTHOR]

Microalgae are photosynthetic microorganisms known for their variety of compounds that can be useful for food, feed, pharmaceuticals, and fuel industries. Microalgae production costs have been one of the major obstacles to large-scale commercial production. Therefore, several studies are still being performed aiming to increase biomass by developing novel photobioreactor design and microalgae cultivation techniques. This work's purpose is to increase Neochloris oleoabundans biomass. Different nutrient-feeding regimes were tested in N. oleoabundans cultivation in Erlenmeyer flasks and bench-scale tubular photobioreactor. In Erlenmeyer flasks, the best concentrations of nitrate and phosphate were 8.82 mM and 5.16 mM, respectively. In bench-scale tubular photobioreactor, NaNO3 proved to be the best nitrogen source, in comparison with (NH4)2SO4 and NH4NO3. Still in the photobioreator, the addition of nitrate by fed-batch process combined with automated system of CO2 feeding showed to be of utmost importance for insuring a high density of N. oleoabundans. The essential nutrients evaluated in this work play an important role on N. oleoabundans biomass accumulation, as well as the photobioreactor configuration and feeding regimes. [ABSTRACT FROM AUTHOR]

Vascular smooth muscle cell (VSMC) function is regulated by Nox-derived reactive oxygen species (ROS) and redox-dependent signaling in discrete cellular compartments. Whether cholesterol-rich microdomains (lipid rafts/caveolae) are involved in these processes is unclear. Here we examined the sub-cellular compartmentalization of Nox isoforms in lipid rafts/caveolae and assessed the role of these microdomains in VSMC ROS production and pro-contractile and growth signaling. Intact small arteries and primary VSMCs from humans were studied. Vessels from Cav-1−/− mice were used to test proof of concept. Human VSMCs express Nox1, Nox4, Nox5 and Cav-1. Cell fractionation studies showed that Nox1 and Nox5 but not Nox4, localize in cholesterol-rich fractions in VSMCs. Angiotensin II (Ang II) stimulation induced trafficking into and out of lipid rafts/caveolae for Nox1 and Nox5 respectively. Co-immunoprecipitation studies showed interactions between Cav-1/Nox1 but not Cav-1/Nox5. Lipid raft/caveolae disruptors (methyl-β-cyclodextrin (MCD) and Nystatin) and Ang II stimulation variably increased O2− generation and phosphorylation of MLC20, Ezrin-Radixin-Moesin (ERM) and p53 but not ERK1/2, effects recapitulated in Cav-1 silenced (siRNA) VSMCs. Nox inhibition prevented Ang II-induced phosphorylation of signaling molecules, specifically, ERK1/2 phosphorylation was attenuated by mellitin (Nox5 inhibitor) and Nox5 siRNA, while p53 phosphorylation was inhibited by NoxA1ds (Nox1 inhibitor). Ang II increased oxidation of DJ1, dual anti-oxidant and signaling molecule, through lipid raft/caveolae-dependent processes. Vessels from Cav-1−/− mice exhibited increased O2− generation and phosphorylation of ERM. We identify an important role for lipid rafts/caveolae that act as signaling platforms for Nox1 and Nox5 but not Nox4, in human VSMCs. Disruption of these microdomains promotes oxidative stress and Nox isoform-specific redox signalling important in vascular dysfunction associated with cardiovascular diseases. [ABSTRACT FROM AUTHOR]

Background and Aim: Escherichia coli is one of the main pathogens responsible for veterinary and human infections, and it is associated with significant economic losses in the livestock, as it causes severe diseases to humans, particularly in children. For that reason, there is a need for introducing new drugs to treat E. coli diseases. The Brazilian species richness is a source of potential new antibacterial natural products. The study aimed at the biological and chemical investigation of the organic extract obtained from the stem of Microplumeria anomala (Apocynaceae), EB127, as it was identified as a potential source of new antibacterial compounds to be used in Veterinary. Materials and Methods: The antibacterial activity was evaluated by disk diffusion and microdilution assays; chromatography, nuclear magnetic resonance spectrometry, and mass spectrometry were used in the isolation and identification of compounds. Results: EB127 showed activity against E. coli ATCC25922, and against three E. coli strains that were isolated from frigarte's cloaca, named 31/1A, 35A, and 51A. Lupeol, 3-acetyl-11-oxo-ß-amyrin, 3-acetyl-11-oxo-a-amyrin, sitosterol, stigmasterol, 3ß,7a-dihydroxy-cholest-5-ene, 3ß-hydroxy-cholest-5-en-7-one, and 3ß-hydroxy-cholest-5,22-dien-7-one were identified in fraction Hex/CHCl3, while loganin, loganic acid, methylanomaline, and anomaline were all identified in EB127 and protocatechuic acid hexoside, ferulic acid, secoxyloganin, feruloylquinic acid, vanillic acid hexoside, protocatechuic acid-4-O-ß-hexoside, and rosmarinic acid were tentatively identified in fraction 10%ACN/H2O. E. coli 51A (virulent/non-resistant) showed sensitivity to the antibacterial action of fraction Hex/CHCl3 which contains alkaloids, triterpenes, and steroids, while E. coli 35A (resistant/non-virulent) were more susceptible to 10%ACN/H2O, which contains iridoids as loganin and loganic acid, and glycosylated and non-glycosylated caffeic acids. Conclusion: Fraction 10%ACN/H2O is of interest in pursuing new drugs to treat resistant E. coli, in veterinary. All compounds were isolated from the plant for the first time and have shown potential as new antibacterial natural products from Amazon plants to be used in veterinary and human diseases. [ABSTRACT FROM AUTHOR]

Environmental impacts can be more accurately assessed with the aid of spatial characterization of potentially toxic elements (PTEs). In fact, developing cost-effective, environmentally friendly spatial characterization methods for PTEs can facilitate the expeditious, accurate, detailed diagnosis of soil in large areas. In this study, we used three geomorphic surfaces of Oxisols to assess the ability of chemical and X-ray diffraction analyses of iron oxides, diffuse reflectance spectroscopy (DRS) and magnetic susceptibility (MS) measurements to predict the contents in PTEs (Ba, Co, Cr, Cu, Mn, Mo, Ni, Pb and Zn) and their spatial variability. Magnetic susceptibility and diffuse reflectance spectroscopy allowed well-calibrated prediction models for Ba, Co, Cu, Mn and Ni to be developed, whereas DRS-calibrated methods afforded more accurate prediction of Ba and Mn contents, and magnetic susceptibility-calibrated methods of Co and Ni contents. The correlation between PTEs and free iron contents, and their spatial pattern, testifies to the goodness of the proposed methods for predicting the contents of potentially toxic elements in soils. [ABSTRACT FROM AUTHOR]

Diffuse reflectance spectroscopy (DRS) provides a rapid, non-destructive, less expensive and sustainable complement to conventional methods of soil analysis. This study aims to assess the DRS ability to predict contents of clay, iron oxide and adsorbed phosphate (Pads), characterizing their spatial variability in Oxisols from Brazil. Soil samples collected from three geomorphic surfaces were scanned in the visible and near infrared spectral range. Moreover, chemometric calibration was obtained through partial least-squares regression (PLSR). Then, spatial distribution maps of the attributes were constructed using predicted values from calibrated models through geostatistical methods. The best accuracy of prediction models (residual prediction deviation > 2.0) for clay, extractable Fe by dithionite–citrate–bicarbonate and hematite was obtained within the visible region (380–800 nm), and for adsorbed phosphate within the visible near infrared (380–2300 nm). Predicted values of these attributes had low error (< 10%) with the exception of goethite, and enabled building maps similar to those with measured values, demonstrating the DRS importance for clay, iron oxide and adsorbed phosphate mapping of large areas and at detailed scale. [ABSTRACT FROM AUTHOR]

Cationic and hydrophilic coatings based on casting and drying water dispersions of two different nanoparticles (NPs) onto glass are here described and evaluated for antimicrobial activity. Discoid cationic bilayer fragments (BF) surrounded by carboxy-methylcellulose (CMC) and poly (diallyl dimethyl ammonium) chloride (PDDA) NPs and spherical gramicidin D (Gr) NPs dispersed in water solution were cast onto glass coverslips and dried, forming a coating quantitatively evaluated against Pseudomonas aeruginosa, Staphylococcus aureus and Candida albicans. From plating and colony forming units (CFU) counting, all strains interacting for 1 h with the coatings lost viability from 105 to 106, to zero CFU, at two sets of Gr and PDDA doses: 4.6 and 25 μg, respectively, or, 0.94 and 5 μg, respectively. Combinations produced broad spectrum, antimicrobial coatings; PDDA electrostatically attached to the microbes damaging cell walls, allowing Gr NPs interaction with the cell membrane. This concerted action promoted optimal activity at low Gr and PDDA doses. Further washing and drying of the deposited dried coatings showed that they were washed out so that antimicrobial activity was no longer present on the glass surface. Significant applications in biomedical materials can be foreseen for these transient coatings. [ABSTRACT FROM AUTHOR]

Abstract: Relief is regarded as the abiotic factor most strongly influencing pedogenic processes at a local scale. The spatial correlations between the composition of the clay fraction (iron — Fe and aluminum — Al oxides, kaolinite and organic matter — OM) and contents of available phosphorus (P) of an Oxisol were evaluated at hillslope scale under sugarcane cultivation. A total of 119 samples were collected at intersection points on a 100×100m georeferenced grid of regularly spaced points 10m apart in the 0.2–0.4m depth in an area consisting of two landform components namely: component I (an area with a linear hillslope curvature), and component II (one with a concave–convex hillslope curvature). Soil OM and available P contents were subjected to descriptive statistics and geostatistical analyses in order to assess their variability and spatial dependence. All attributes studied were spatially dependent. Available phosphorus had positive spatial correlation with high crystalline goethite, hematite and gibbsite. Identifying small hillslope curvatures is useful with a view to better understanding their relationships with soil organic matter and available phosphorus, as well as kaolinite and Fe and Al oxide attributes. A simple correlation analysis by itself is inadequate to relate attributes, which requires a supplemental, geostatistical technique. [Copyright &y& Elsevier]

R3398 --> Introduction: COVID‐19 is primarily a respiratory disease associated with cardiovascular risk. SARS‐CoV‐2, the virus causing COVID‐19, uses ACE2, an important enzyme in the cardiovascular system that regulates the conversion of Ang II (deleterious/pro‐hypertensive) to Ang 1‐7 (protective/anti‐hypertensive), as a receptor for host cell entry and infection. Considering the relationship between the viral S1‐protein and the host's ACE2, it is unclear whether this interaction is merely a mechanism of infection or whether it also contributes to cardiovascular damage associated with COVID‐19. We hypothesisedthat SARS‐Cov‐2‐ACE2 interaction induces activation of vascular cell inflammatory responses that are influenced by ACE2 dependent and/or independent enzymatic Ang‐(1‐7) production. Methods: Human microvascular endothelial cells (MEC) were used and stimulated with SARS‐CoV‐2 recombinant S1 protein (rS1p) (0.66 μg/mL) at 10/30 min (acute) and 5/24h (chronic). Activation of pro‐inflammatory signaling pathways (immunoblotting, real‐time PCR), microparticle (MP) generation (NanoSight), and cytokine production (ELISA) were assessed. In some experiments, cells were pre‐incubated with an ACE2 activator (DIZE – 190 nM) and inhibitor (MLN‐4760 – 440 pM). Results: rS1P increased NFκB activation (Control ©=0.99±0.06 vs. 1.38±0.19 AU; p<0.05) and MP formation (C=1.01±0.17 vs. 2.06±0.21, x109/mL; p<0.05), a marker of endothelial cell damage. mRA expression of IL‐1β (C=1.07±0.13 vs. 50.04±4.63 2^‐ddCT), IL6 (C=1.15±0.0.19 vs. 13.52±2 2^‐ddCT), TNFα (C=1.12±0.16 vs. 20.29±2.15 2^‐ddCT), VCAM‐1 (C=1.21±0.24 vs. 13.39±1.57 2^‐ddCT) and MCP‐1 (C=1.02±0.06 vs. 7.01±2.16 2^‐ddCT) was increased by rS1p (p<0.05). This was associated with an increased production of IL6 (C=22.77±3.27 vs. 1221±18.33 pg/mL) and MCP‐1 (C=876.9±33.47 vs. 1110±13.33 pg/mL, (p<0.05). rS1p did not change gene levels of TGFβ, galectin‐3 or ACE2. While DIZE did not influence rS1p pro‐inflammatory markers; MLN‐4760 potentiated rS1p‐induced increase in IL‐1β gene levels (rS1p=27.59±5.05 vs. MLN=62.11±11.17 2^‐ddCT) and blocked rS1p effects on MCP‐1 (rS1p=5.08±0.47 vs. MLN=2.43±0.6 2^‐ddCT) and VCAM‐1 (rS1p=72.82±13.76 vs. MLN=20.74±4.53 2^‐ddCT). ACE2 inhibition did not interfere with rS1p‐induced increase in IL6 or TNFα mRNA expression. Analysis of MP content revealed the expression of ACE2 short form (60 kDa) and flotillin‐1, suggesting that ACE2 localizes in membrane microdomains. Conclusions: Our data demonstrate that in human endotheial cells rS1P induces a pro‐inflammatory response and stimulates generation of ACE2‐containing microparticles. These findings suggest that the spike protein 1 of SARS‐CoV‐2 has a direct injurious effect on the endothelium that may contribute to endotheliitis in COVID‐19. Whether these processes are associated with viral infection await clarification. [ABSTRACT FROM AUTHOR]

Inflammatory arthritis, such as rheumatoid arthritis (RA), stands out as one of the main sources of pain and impairment to the quality of life. The use of hemopressin (PVNFKFLSH; Hp), an inverse agonist of type 1 cannabinoid receptor, has proven to be effective in producing analgesia in pain models, but its effect on neuro-inflammatory aspects of RA is limited. In this study, antigen-induced arthritis (AIA) was evoked by the intraarticular (i.art.) injection of methylated bovine serum albumin (mBSA) in male Sprague Dawley rats. Phosphate buffered saline (PBS)-injected ipsilateral knee joints or AIA contralateral were used as control. Nociceptive and inflammatory parameters such as knee joint oedema and leukocyte influx and histopathological changes were carried out in addition to the local measurement of interleukins (IL) IL-6, IL-1β, tumor necrosis factor-α and the immunoreactivity of the neuropeptides substance P (SP) and calcitonin gene related peptide (CGRP) in the spinal cord (lumbar L3-5 segments) of AIA rats. For 4 days, AIA rats were treated daily with a single administration of saline, Hp injected (10 or 20 μg/day, i.art.), Hp given orally (20 μg/Kg, p.o.) or indomethacin (Indo; 5 mg/Kg, i.p.). In comparison to the PBS control group, the induction of AIA produced a significant and progressive mono-arthritis condition. The degree of AIA severity progressively compromised the normal walking pattern and impaired mobility over the next four days in relation to PBS-injected rats or contralateral knee joints. In AIA rats, the reduction of the distance between footprints and disturbances of gait evidenced signs of nociception. This response worsened at day 4, and a loss of footprint from the ipsilateral hind paw was evident. Daily treatment of the animals with Hp either i.art. (10 and 20 μg/knee) or p.o. (20 μg/Kg) as well as Indo (5 mg/Kg, i.p.) ameliorated the impaired mobility in a time-dependent manner (P < 0.05). In parallel, the AIA-injected ipsilateral knee joints reach a peak of swelling 24 h after AIA induction, which persisted over the next four days in relation to PBS-injected rats or contralateral knee joints. There was a significant but not dose-dependent inhibitory effect produced by all dosages and routes of Hp treatments on AIA-induced knee joint swelling (P < 0.05). In addition, the increased synovial levels of MPO activity, total leukocytes number and IL-6, but not IL-1β, were significantly reduced by the lower i.art. dose of Hp. In conclusion, these results successfully demonstrate that Hp may represent a novel therapeutic strategy to treat RA, an effect which is unrelated to the proinflammatory actions of the neuropeptides CGRP and SP. [ABSTRACT FROM AUTHOR]

The green microalga Chlamydomonas reinhardtii is a model microorganism for several areas of study. Among the different microalgae species, it presents advantageous characteristics, such as genomes completely sequenced and well‐established techniques for genetic transformation. Despite that, C. reinhardtii production is still not easily commercially viable, especially due to the low biomass yield. So far there are no reports of scientometric study focusing only on C. reinhardtii biomass production process. Considering the need for culture optimization, a scientometric research was conducted to analyze the papers that investigated the growth regimes effects in C. reinhardtii cultivation. The search resulted in 130 papers indexed on Web of Science and Scopus platforms from 1969 to December 2022. The quantitative analysis indicated that the photoautotrophic regime was the most employed in the papers. However, when comparing the three growth regimes, the mixotrophic one led to the highest production of biomass, lipids, and heterologous protein. The production of bioproducts was considered the main objective of most of the papers and, among them, biomass was the most frequently investigated. The highest biomass production reported among the papers was 40 g L−1 in the heterotrophic growth of a transgenic strain. Other culture conditions were also crucial for C. reinhardtii growth, for instance, temperature and cultivation process. [ABSTRACT FROM AUTHOR]

Significance: NADPH oxidases (Noxs), of which there are seven isoforms (Nox1–5, Duox1/Duox2), are professional oxidases functioning as reactive oxygen species (ROS)-generating enzymes. ROS are signaling molecules important in physiological processes. Increased ROS production and altered redox signaling in the vascular system have been implicated in the pathophysiology of cardiovascular diseases, including hypertension, and have been attributed, in part, to increased Nox activity. Recent Advances: Nox1, Nox2, Nox4, and Nox5 are expressed and functionally active in human vascular cells. While Nox1, Nox2, and Nox4 have been well characterized in models of cardiovascular disease, little is known about Nox5. This may relate to the lack of experimental models because rodents lack NOX5. However, recent studies have advanced the field by (i) elucidating mechanisms of Nox5 regulation, (ii) identifying Nox5 variants, (iii) characterizing Nox5 expression, and (iv) discovering the Nox5 crystal structure. Moreover, studies in human Nox5-expressing mice have highlighted a putative role for Nox5 in cardiovascular disease. Critical Issues: Although growing evidence indicates a role for Nox-derived ROS in cardiovascular (patho)physiology, the exact function of each isoform remains unclear. This is especially true for Nox5. Future Directions: Future directions should focus on clinically relevant studies to discover the functional significance of Noxs, and Nox5 in particular, in human health and disease. Two important recent studies will impact future directions. First, Nox5 is the first Nox to be crystallized. Second, a genome-wide association study identified Nox5 as a novel blood pressure-associated gene. These discoveries, together with advancements in Nox5 biology and biochemistry, will facilitate discovery of drugs that selectively target Noxs to interfere in uncontrolled ROS generation. [ABSTRACT FROM AUTHOR]

This review aimed to present Chlamydomonas reinhardtii as an alternative for heterologous protein production, especially for biopharmaceuticals, and its general characteristics when compared with other expression systems. The need to produce heterologous proteins for industrial interest, therapeutic ends, and diagnostic kits has led to the development of recombinant microalgal technology. This technology presents some interesting features, such as rapid growth and low transgene dispersion compared to plants, the ability to fold complex proteins compared to bacteria, and low production costs compared to other expression systems, such as yeast and mammalian cells. Overall, C. reinhardtii heterologous protein expression is coming of age with several research groups focused on developing an optimal producer strain. [ABSTRACT FROM AUTHOR]

Hyperaldosteronism causes cardiovascular disease as well as hypomagnesemia. Mechanisms are ill-defined but dysregulation of TRPM7, a Mg2+-permeable channel/α-kinase, may be important. We examined the role of TRPM7 in aldosterone-dependent cardiovascular and renal injury by studying aldosterone-salt treated TRPM7-deficient (TRPM7+/Δkinase) mice. Plasma/tissue [Mg2+] and TRPM7 phosphorylation were reduced in vehicle-treated TRPM7+/Δkinase mice, effects recapitulated in aldosterone-salt-treated wild-type mice. Aldosterone-salt treatment exaggerated vascular dysfunction and amplified cardiovascular and renal fibrosis, with associated increased blood pressure in TRPM7+/Δkinase mice. Tissue expression of Mg2+-regulated phosphatases (PPM1A, PTEN) was downregulated and phosphorylation of Smad3, ERK1/2, and Stat1 was upregulated in aldosterone-salt TRPM7-deficient mice. Aldosterone-induced phosphorylation of pro-fibrotic signaling was increased in TRPM7+/Δkinase fibroblasts, effects ameliorated by Mg2+ supplementation. TRPM7 deficiency amplifies aldosterone-salt-induced cardiovascular remodeling and damage. We identify TRPM7 downregulation and associated hypomagnesemia as putative molecular mechanisms underlying deleterious cardiovascular and renal effects of hyperaldosteronism. Deficiency of the Mg2+-permeable channel/α-kinase TRPM7 in mice increases susceptibility to cardiovascular and renal fibrosis induced by aldosterone and salt. [ABSTRACT FROM AUTHOR]

R2703 --> Introduction: Interferon (IFN) alpha (IFNα) and lambda3 (IFNλ3) constitute the first line of immunity against viral infection by increasing expression of interferon‐stimulated genes (ISGs). Both are produced in SARS‐CoV‐2 infection and have been considered therapeutically in COVID‐19. In epithelial cells IFNs have been shown to influence the expression of angiotensin‐converting enzyme 2 (ACE2), the receptor for S‐protein (S1P) of SARS‐CoV‐2. ACE2 is part of the vascular renin angiotensin system and may be important in COVID‐19‐associated endotheliitis. Whether SP1 influences immune responses in endothelial cells unknown. We investigated effects of SP1 on ACE2 expression and immune responses in human endothelial cells. Methods: Culturedhuman Microvascular, Lymphatic, Aortic and Pulmonary Endothelial Cells (MEC, LEC, AEC, and PEC respectively) were stimulated with SP1 of SARS‐CoV‐2 (1µg/106 cells), IFNα (100ng/mL) or IFNλ3 (100IU/mL). Because ACE2, metalloproteinase domain 17 (ADAM17) and type II transmembrane serine protease (TMPRSS2) are important for SARS‐CoV‐2 infection, cells were treated with inhibitors of ADAM17 (marimastat, 3.8nM and TAPI‐1, 100nM), ACE2 (MLN4760, 440pM), and TMPRSS2 (camostat, 50µM). Expression of ISGs (ISG15, IFIT1, and MX1) was investigated by real‐time PCR (5h), cytokine by ELISA and protein expression by immunoblotting (24h). Results: AEC, LEC, MEC, and PEC stimulated with SP1 exhibited increased expression of ISGs: ISG15 (1.9‐2.3‐fold), IFIT1 (2.0‐7.9‐fold), MX1 (3.1‐5.8‐fold), indicating activation of the IFN pathway. MEC and LEC exhibited higher responses to IFNα activation. MEC (ISG15: 16‐fold, IFIT1: 21‐fold, MX1: 31‐fold) and LEC (ISG15: 48‐fold, IFIT1: 105‐fold, MX1: 134‐fold). Only MEC exhibited ISG expression induced by IFNλ3 (ISG15: 1.7‐fold, IFIT1: 1.9‐fold, MX1: 1.7‐fold) (P<0.05). IL‐6 production was increased in MEC stimulated with IFNα (1230pg/mL) and IFNλ3 (1124pg/mL) vs control (591pg/mL). Marimastat, but not camostat or MLN4760 inhibited SP1 effects. IFNα induced IL‐6 production in AEC and PEC (178pg/mL vs control 109pg/mL). No changes in IL‐6 were observed in LEC. IFNα increased expression of the short form of ACE2 (75kDa) (63%), ADAM17 (36%), and TMPRSS2 (65%) in MECs. This was associated with increased activation of pro‐inflammatory pathways with increased phosphorylation of Stat1 (134%), Stat2 (102%), ERK1/2 (42%). IFNλ3 also increased phosphorylation of Stat2 (60%) and ERK1/2 (75%). In MEC, phosphorylation of the eNOS activation motif Ser1177 was reduced by IFNLα and (40%) and IFNλ3 (40%). Conclusions: In human endothelial cells from multiple vascular beds, SP1, IFNα and IFNλ3 induced an immune response characterised by increased ISG expression and IL‐6 production, processes that involve ADAM17. These effects were associated with increased expression of the short form of ACE2 and activation of pro‐inflammatory pathways. Our novel findings demonstrate that SP‐1 induces an endothelial immune and inflammatory response that may be important in endotheliitis associated with COVID‐19. [ABSTRACT FROM AUTHOR]

R3405 --> Introduction: Mice expressing human Nox5 (hNox5) in vascular smooth muscle cells (VSMC) exhibit increased contraction, ROS generation, cardiac fibrosis and increased pulse pressure. In kidneys, hNox5 in podocytes is associated with albuminuria, hypertension and inflammation. Here we questioned whether VSMC Nox5 influences kidney function and damage in Ang II‐induced hypertension. Methods: Wildtype (WT) and mice expressing hNOX5 in SMC (Nox5+SM22+) were studied. Mice were infused with Ang II (600 ng/Kg/day) for 28 days. Blood pressure was assessed by tail‐cuff, kidney damage by histology, and mRNA and protein expression by qPCR and western‐blotting, respectively. Inflammatory infiltrate was assessed by FACS. Results: Ang II increased blood pressure (mmHg) in WT (162.7±9) and Nox5+SM22+ (162.3±10). Albuminuria was increased in Nox5+SM22+ versus WT mice (10.2±6 vs 1.27±0.3 mg/mmol, p<0.05) and by Ang II in both groups (Nox5: 44.8±18; WT: 58.7±12 mg/mmol). Kidneys from Ang II‐infused Nox5+SM22+ mice exhibited significant perivascular fibrosis and inflammatory cell infiltration compared to WT. Kidney expression of vimentin (AU: 1.01±0.05 vs WT 0.85±0.03) and αSMA (AU: 0.44±0.03 vs WT 0.33±0.01), markers of myofibroblast differentiation, was increased in Nox5 mice (p<0.05), an effect that was augmented by Ang II. In kidneys from Nox5+SM22+ mice, levels of macrophage F4/80+ cells were increased (%: 24±2 vs WT 18±1, p<0.05). Ang II increased macrophage homing in kidneys from WT mice (34±3%, p<0.05 vs non‐treated), but did not further increase it in Nox5 mice (35±5%). Levels of cytotoxic CD8+ T cells and expression of IL‐1b and IRAK were increased by Ang II in Nox5 mice. Conclusion: Our study suggests that Nox5 in VSMCs induces renal fibrosis and pro‐inflammatory signalling by Ang II, likely through increased ROS generation. [ABSTRACT FROM AUTHOR]

Objective: Transient receptor potential (TRP) melastatin 7 (TRPM7) cation channel, a dual-function ion channel/protein kinase, regulates vascular smooth muscle cell (VSMC) Mg2+ homeostasis and mitogenic signaling. Mechanisms regulating vascular growth effects of TRPM7 are unclear, but epidermal growth factor (EGF) may be important because it is a magnesiotropic hormone involved in cellular Mg2+ regulation and VSMC proliferation. Here we sought to determine whether TRPM7 is a downstream target of EGF in VSMCs and if EGF receptor (EGFR) through TRPM7 influences VSMC function. Approach and results: Studies were performed in primary culture VSMCs from rats and humans and vascular tissue from mice deficient in TRPM7 (TRPM7+/kinase and TRPM7R/R). EGF increased expression and phosphorylation of TRPM7 and stimulated Mg2+ influx in VSMCs, responses that were attenuated by gefitinib (EGFR inhibitor) and NS8593 (TRPM7 inhibitor). Co-immunoprecipitation (IP) studies, proximity ligation assay (PLA) and live-cell imaging demonstrated interaction of EGFR and TRPM7, which was enhanced by EGF. PP2 (c-Src inhibitor) decreased EGF-induced TRPM7 activation and prevented EGFR-TRPM7 association. EGF-stimulated migration and proliferation of VSMCs were inhibited by gefitinib, PP2, NS8593 and PD98059 (ERK1/2 inhibitor). Phosphorylation of EGFR and ERK1/2 was reduced in VSMCs from TRPM7+/kinase mice, which exhibited reduced aortic wall thickness and decreased expression of PCNA and Notch 3, findings recapitulated in TRPM7R/R mice. Conclusions: We show that EGFR directly interacts with TRPM7 through c-Src-dependent processes. Functionally these phenomena regulate [Mg2+]i homeostasis, ERK1/2 signaling and VSMC function. Our findings define a novel signaling cascade linking EGF/EGFR and TRPM7, important in vascular homeostasis. [ABSTRACT FROM AUTHOR]

Aims Transient Receptor Potential Melastatin 7 (TRPM7) cation channel is a chanzyme (channel + kinase) that influences cellular Mg2+ homeostasis and vascular signalling. However, the pathophysiological significance of TRPM7 in the cardiovascular system is unclear. The aim of this study was to investigate the role of this chanzyme in the cardiovascular system focusing on inflammation and fibrosis. Methods and results TRPM7-deficient mice with deletion of the kinase domain (TRPM7+/Δkinase) were studied and molecular mechanisms investigated in TRPM7+/Δkinase bone marrow-derived macrophages (BMDM) and co-culture systems with cardiac fibroblasts. TRPM7-deficient mice had significant cardiac hypertrophy, fibrosis, and inflammation. Cardiac collagen and fibronectin content, expression of pro-inflammatory mediators (SMAD3, TGFβ) and cytokines [interleukin (IL)-6, IL-10, IL-12, tumour necrosis factor-α] and phosphorylation of the pro-inflammatory signalling molecule Stat1, were increased in TRPM7+/Δkinase mice. These processes were associated with infiltration of inflammatory cells (F4/80+CD206+ cardiac macrophages) and increased galectin-3 expression. Cardiac [Mg2+]i, but not [Ca2+]i, was reduced in TRPM7+/Δkinase mice. Calpain, a downstream TRPM7 target, was upregulated (increased expression and activation) in TRPM7+/Δkinase hearts. Vascular functional and inflammatory responses, assessed in vivo by intra-vital microscopy, demonstrated impaired neutrophil rolling, increased neutrophil: endothelial attachment and transmigration of leucocytes in TRPM7+/Δkinase mice. TRPM7+/Δkinase BMDMs had increased levels of galectin-3, IL-10, and IL-6. In co-culture systems, TRPM7+/Δkinase macrophages increased expression of fibronectin, proliferating cell nuclear antigen, and TGFβ in cardiac fibroblasts from wild-type mice, effects ameliorated by MgCl2 treatment. Conclusions We identify a novel anti-inflammatory and anti-fibrotic role for TRPM7 and suggest that its protective effects are mediated, in part, through Mg2+-sensitive processes. [ABSTRACT FROM AUTHOR]

New Findings: What is the topic of this review?This review provides a comprehensive overview of Nox5 from basic biology to human disease and highlights unique features of this Nox isoformWhat advances does it highlight?Major advances in Nox5 biology relate to crystallization of the molecule and new insights into the pathophysiological role of Nox5. Recent discoveries have unravelled the crystal structure of Nox5, the first Nox isoform to be crystalized. This provides new opportunities to develop drugs or small molecules targeted to Nox5 in an isoform‐specific manner, possibly for therapeutic use. Moreover genome wide association studies (GWAS) identified Nox5 as a new blood pressure‐associated gene and studies in mice expressing human Nox5 in a cell‐specific manner have provided new information about the (patho) physiological role of Nox5 in the cardiovascular system and kidneys. Nox5 seems to be important in the regulation of vascular contraction and kidney function. In cardiovascular disease and diabetic nephropathy, Nox5 activity is increased and this is associated with increased production of reactive oxygen species and oxidative stress implicated in tissue damage. Nicotinamide adenine dinucleotide phosphate (NADPH) oxidases (Nox), comprise seven family members (Nox1–Nox5 and dual oxidase 1 and 2) and are major producers of reactive oxygen species in mammalian cells. Reactive oxygen species are crucially involved in cell signalling and function. All Noxs share structural homology comprising six transmembrane domains with two haem‐binding regions and an NADPH‐binding region on the intracellular C‐terminus, whereas their regulatory systems, mechanisms of activation and tissue distribution differ. This explains the diverse function of Noxs. Of the Noxs, NOX5 is unique in that rodents lack the gene, it is regulated by Ca2+, it does not require NADPH oxidase subunits for its activation, and it is not glycosylated. NOX5 localizes in the perinuclear and endoplasmic reticulum regions of cells and traffics to the cell membrane upon activation. It is tightly regulated through numerous post‐translational modifications and is activated by vasoactive agents, growth factors and pro‐inflammatory cytokines. The exact pathophysiological significance of NOX5 remains unclear, but it seems to be important in the physiological regulation of sperm motility, vascular contraction and lymphocyte differentiation, and NOX5 hyperactivation has been implicated in cardiovascular disease, kidney injury and cancer. The field of NOX5 biology is still in its infancy, but with new insights into its biochemistry and cellular regulation, discovery of the NOX5 crystal structure and genome‐wide association studies implicating NOX5 in disease, the time is now ripe to advance NOX5 research. This review provides a comprehensive overview of our current understanding of NOX5, from basic biology to human disease, and highlights the unique characteristics of this enigmatic Nox isoform. [ABSTRACT FROM AUTHOR]

Hypertension is a major risk factor for many common chronic diseases, such as heart failure, myocardial infarction, stroke, vascular dementia, and chronic kidney disease. Pathophysiological mechanisms contributing to the development of hypertension include increased vascular resistance, determined in large part by reduced vascular diameter due to increased vascular contraction and arterial remodelling. These processes are regulated by complexinteracting systems such as the renin-angiotensin-aldosterone system, sympathetic nervous system, immune activation, and oxidative stress, which influence vascular smooth muscle function. Vascular smooth muscle cells are highly plastic and in pathological conditions undergo phenotypic changes from a contractile to a proliferative state. Vascular smooth muscle contraction is triggered by an increase in intracellular free calcium concentration ([Ca2+]i), promoting actin-myosin cross-bridge formation. Growing evidence indicates that contraction is also regulated by calcium-independent mechanisms involving RhoA-Rho kinase, protein Kinase C and mitogen-activated protein kinase signalling, reactive oxygen species, and reorganization of the actin cytoskeleton. Activation of immune/inflammatory pathways and non-coding RNAs are also emerging as important regulators of vascular function. Vascular smooth muscle cell [Ca2+]i not only determines the contractile state but also influences activity of many calciumdependent transcription factors and proteins thereby impacting the cellular phenotype and function. Perturbations in vascular smooth muscle cell signalling and altered function influence vascular reactivity and tone, important determinants of vascular resistance and blood pressure. Here, we discuss mechanisms regulating vascular reactivity and contraction in physiological and pathophysiological conditions and highlight some new advances in the field, focusing specifically on hypertension. [ABSTRACT FROM AUTHOR]

The development of fast, accurate and low-cost methods to quantify soil attributes is of paramount importance to enable detailed mapping, mainly in tropical regions where there is great variation of the chemical, physical and mineralogical attributes. Therefore, the aims of this paper were (i) to investigate if visible and near infrared (VIS-NIR) spectroscopy and magnetic susceptibility (MS) can be applied to determine soil attributes at the sandstone-basaltic transition and (ii) evaluate and map their spatial distribution. Calibration models based on VIS-NIR spectroscopy and MS were developed separately for each attribute. Soil samples (0-25 cm depth) were collected at 446 sites, air-dried and passed through a 2-mm sieve and analyzed in the laboratory. To develop models based on soil spectra and laboratory data, the partial least squares regression (PLSR) was used. Already, the MS-based models were calibrated by linear regression between magnetic and laboratory data. The best prediction accuracy parameters were obtained with MS, later with VIS-NIR and lastly with VIS. The more accurate results between the observed and predicted values were found for iron oxide extracted by dithionite (R² = 0.89, RRMSE = 0.02), clay (R² = 0.85, RRMSE = 0.76) and total carbon (R² = 0.83, RRMSE = 1.18) estimated by MS, revealing that this is a good predictor of key properties of studied soils, even with wide chemical and mineralogical variation. Both tools are very attractive for the strategic planning of land use and occupation, mapping large areas with detailed scale, environmental monitoring and precision agriculture. [ABSTRACT FROM AUTHOR]

Highlights: [•] Hematite, goethite and dithionite iron exhibited well-defined anisotropy. [•] Preparation and management practices soil affected soil CO2 emission. [•] The minerals can auxiliary in anisotropy identify the soil CO2 emission. [Copyright &y& Elsevier]

Significance: Reactive oxygen species (ROS) are produced during normal endoplasmic reticulum (ER) metabolism. There is accumulating evidence showing that under stress conditions such as ER stress, ROS production is increased via enzymes of the NADPH oxidase (Nox) family, especially via the Nox2 and Nox4 isoforms, which are involved in the regulation of blood pressure. Hypertension is a major contributor to cardiovascular and renal disease, and it has a complex pathophysiology involving the heart, kidney, brain, vessels, and immune system. ER stress activates the unfolded protein response (UPR) signaling pathway that has prosurvival and proapoptotic components. Recent Advances: Here, we summarize the evidence regarding the association of Nox enzymes and ER stress, and its potential contribution in the setting of hypertension, including the role of other conditions that can lead to hypertension ( e.g., insulin resistance and diabetes). Critical Issues: A better understanding of this association is currently of great interest, as it will provide further insights into the cellular mechanisms that can drive the ER stress-induced adaptive versus maladaptive pathways linked to hypertension and other cardiovascular conditions. More needs to be learnt about the precise signaling regulation of Nox(es) and ER stress in the cardiovascular system. Future Directions: The development of specific approaches that target individual Nox isoforms and the UPR signaling pathway may be important for the achievement of therapeutic efficacy in hypertension. Antioxid. Redox Signal. 20, 121-134. [ABSTRACT FROM AUTHOR]

Predicting and mapping productivity areas allows crop producers to improve their planning of agricultural activities. The primary aims of this work were the identification and mapping of specific management areas allowing coffee bean quality to be predicted from soil attributes and their relationships to relief. The study area was located in the Southeast of the Minas Gerais state, Brazil. A grid containing a total of 145 uniformly spaced nodes 50 m apart was established over an area of 31.7 ha from which samples were collected at depths of 0.00-0.20 m in order to determine physical and chemical attributes of the soil. These data were analysed in conjunction with plant attributes including production, proportion of beans retained by different sieves and drink quality. The results of principal component analysis (PCA) in combination with geostatistical data showed the attributes clay content and available iron to be the best choices for identifying four crop production environments. Environment A, which exhibited high clay and available iron contents, and low pH and base saturation, was that providing the highest yield (30.4l ha) and best coffee beverage quality (61 sacks ha). Based on the results, we believe that multivariate analysis, geostatistics and the soil-relief relationships contained in the digital elevation model (DEM) can be effectively used in combination for the hybrid mapping of areas of varying suitability for coffee production. [ABSTRACT FROM AUTHOR]

R2706 --> Introduction: : TRPM7 is a channel permeable to Mg2+, Ca2+ and Zn2+ bound to an alpha‐kinase domain with essential role in cation homeostasis and cell cycle and activation. Hyperaldosteronism, which induces hypertension and cardiovascular fibrosis, is associated with Mg2+ wasting. Here we investigate the importance of TRPM7 in hypertension and fibrosis induced by aldosterone and salt. Methods: Wild‐type (WT) and TRPM7‐deficient (M7+/Δ) mice were treated with aldosterone (600µg/Kg/day) and/or 1% NaCl (drinking water) (aldo, salt or aldo‐salt) for 4 weeks. Blood pressure (BP) was evaluated by tail‐cuff. Vessel structure was assessed by pressure myography. Inflammatory cell infiltrate was investigated by flow cytometry. Molecular mechanisms were investigated in cardiac fibroblasts (CF) from WT and M7+/Δ mice. Protein expression was assessed by western‐blot and histology. Results: M7+/Δ mice exhibited reduced TRPM7 expression (30%) and phosphorylation (62%), levels that were recapitulated in WT mice treated with aldo‐salt. M7+/Δ exhibited increased BP by aldo, salt and aldo‐salt (135‐140mmHg) vs M7+/Δ veh (117mmHg) (p<0.05), whereas in WT, BP was increased only in the aldo‐salt group (134mmHg). Mesenteric arteries from M7+/Δ aldo‐salt exhibited thinner walls by reducing cross‐sectional area (35%) vs WT aldo‐salt (p<0.05). Aldo‐salt induced greater collagen deposition in hearts (68%), kidneys (126%) and aortas (45%) from M7+/Δ vs WT. In kidneys from WT mice, aldo‐salt increased frequency of macrophages (47% vs 32% WT veh) and CD4+T cells (33% vs 22% WT veh) (p<0.05), effects observed already in M7+/Δ veh (macrophages: 42%, CD4+T cells: 34%). Aldosterone increased CD8+T cells in kidneys from M7+/Δ (16% vs M7+/Δ‐veh 9%). Hearts from M7+/Δ veh exhibited increased TGFβ, p‐Smad3 and p‐Stat1 (1.5‐fold) whereas in WT these effects were only found after aldo‐salt. Cardiac expression of PPM1A, a Mg2+‐dependent phosphatase, was reduced (3‐fold) only in M7+/Δ mice. CF from M7+/Δ mice showed reduced of proliferation (30%) and PPM1A (4‐fold) and increased expression of TGFβ, IL‐11, IL‐6 (2‐6‐fold), p‐Stat1 (2‐fold), p‐Smad3 (9‐fold) and p‐ERK1/2 (8‐fold) compared with WT. Mg2+ supplementation normalized cell proliferation and reduced protein phosphorylation in M7+/Δ CF (p<0.05. Conclusions: Our findings identify a protective role of TRPM7 in aldosterone‐salt induced cardiovascular injury, which when downregulated, facilitates hypertension, vascular remodeling and cardiac fibrosis through Mg2+ ‐dependent mechanisms. [ABSTRACT FROM AUTHOR]

• Topographic attributes are an excellent alternative to establish fertility zones. • Topographic attributes can be used to guide soil characterizations. • Concave areas showed higher levels of OM, SB and CEC. • Soil acidity management should be more rigorous in convergent areas. • Regardless of the site, the spatial patterns were temporally maintained. Landscapes are intrinsically related to spatial soil variability. Understanding soil chemical fertility based on topographic parameters is essential to ensuring sustainable agronomic management through the rational use of inputs. The aim of this study was to perform a comprehensive assessment of soil organic matter (OM), pH, sum of bases (SB) and cation exchange capacity (CEC) according to topographic parameters, with the goal of identifying spatial patterns and better understanding the relationship between them to capture the soil variability in sugarcane fields. A soil dataset from nine sugarcane experimental fields was evaluated. Approximately 3000 soil samples collected between 2008 and 2017 were evaluated. The topographic parameters of vertical and horizontal curvatures were related to the variability in the soil attributes. The results showed that the horizontal morphometric classes (H Conv , H Plan and H Div) associated with vertical concave areas (V Conc) presented higher levels of OM, SB and CEC, which indicated that these areas have higher soil chemical fertility. The V Conc H Div showed the highest soil chemical fertility; regardless of site, topographic level and inherent soil fertility. For all vertical morphometric classes (V Conc , V Ret and V Conv), soil pH levels were higher in horizontal divergent areas (H Div) and lower in convergent areas (H Conv), suggesting that stricter soil acidity management was needed in the H Conv areas. The V Conv H Conv areas, where lower soil chemical fertility was observed, should be sampled with greater accuracy for adequate spatial soil characterization because it had a higher coefficient of variation (CV) than the other morphometric classes. The results showed that the detected spatial patterns were temporally stable. With high spatial and temporal stability, topographic parameters could be excellent sources of economically feasible and easily assessed information to support farmers in soil chemical fertility characterization. Finally, topographic attributes proved to be an excellent source about soil spatial variability for farmers to use in their fields aiming a site-specific management or an additional layer of data for management zones delineation. [ABSTRACT FROM AUTHOR]