Clustering of risk factors in hypertensive insulin-dependent diabetics with high sodium-lithium countertransport

Clustering of risk factors in hypertensive insulin-dependent diabetics with high sodium-lithium countertransport. Diabetic nephropathy is more common in patients with a positive family history of hypertension and with elevated red blood cell sodium-lithium countertransport, a marker of risk for essential hypertension. To evaluate whether there is a relationship between this cation transport system and indicators of risk of renal and cardiovascular complications in diabetic patients before the development of clinical proteinuria, we studied 31 type 1 (insulin-dependent) diabetic patients with arterial hypertension, without clinical proteinuria and 12 normotensive normoalbuminuric diabetic patients. Sodium-lithium countertransport activity was significantly higher in hypertensive patients (0.43 ± 0.03 mmol/1 RBC x hr) than in normotensive patients (0.23 ± 0.03; P < 0.001). To better explore the nature of the association between this transport system and arterial hypertension, hypertensive patients were divided in two groups, with high (>0.41 mmol/1 RBC x hr) or normal (<0.41) sodium-lithium countertransport activity. The two groups of hypertensive diabetics were similar in age, sex, body mass index and blood pressure levels. Hypertensive patients with elevated rates of sodium-lithium counter-transport compared with those with normal sodium-lithium counter-transport activity showed elevated glomerular filtration rate (130 ± 4 ml/min/1.73 m2 vs. 122 ± 3; P < 0.05), albumin excretion rate (median 26 /Lcg/min vs. 11; P < 0.001), higher fractional proximal sodium reabsorption (74 ± 1.2% vs. 71.6 ± 0.9; P < 0.01), exchangeable sodium pool (2937 ± 62 mmol/1.73 m2 vs. 2767 ± 56; P < 0.01), larger kidney volume (317 ± 7 ml/1.73 m2 vs. 270 ± 8; P < 0.05) and left ventricular mass index (122 ± 4 g/m2 vs. 107 ± 5; P < 0.05). Hypertensive patients with normal sodium-lithium countertransport activity had renal parameters similar to normotensive diabetic patients, except higher left ventricular mass index and kidney volume. Hypertensive diabetic patients with elevated sodium-lithium countertransport activity also had higher levels of plasma triglycerides, lower plasma HDL-cholesterol and impaired insulin sensitivity (assessed by euglyce-mic insulin-glucose clamp) compared with the other two groups. In conclusion, renal, cardiac and metabolic abnormalities are prominent in hypertensive type 1 (insulin-dependent) diabetic patients with higher sodium-lithium countertransport

Evaluation of the purity of magnesium hydroxide recovered from saltwork bitterns

Magnesium has been listed among the 30 critical raw materials by the European Union. In recent years, many green and sustainable alternative Mg2+ sources have been sought to satisfy the EU’s demand and to avoid mineral ore consumption. In this context, saltwork bitterns, the by-products of solar sea salt production, have attracted much attention thanks to their high Mg2+ concentrations (up to 80 g/L) and low Ca2+ and bicarbonate contents (95% (w/w). This work presents a comprehensive experimental effort of reactive precipitation tests with NaOH solutions at stoichiometric and over-stoichiometric concentrations to: (i) assess the technical feasibility of Mg2+ recovery from real bitterns collected in saltworks of the Trapani district (Italy) and, (ii) for the first time, conduct an extensive purity investigation of the precipitated magnesium hydroxide powders as brucite. This experimental investigation demonstrates the possibility of extracting highly valuable compounds from saltwork bittern waste, embracing the water valorization and resource recovery approach.Peer ReviewedPostprint (published version

Nanostructured Electrochemical Immunosensor based on Gold Nanowires for the Detection of Model Protein

Nowadays the demand for new devices capable of accurate, fast, and in situ real-time analyses is growing rapidly in the medical field. These devices could have various applications, such as clinical diagnosis and remote patient monitoring. In this work, an electrochemical immunosensor based on Gold Nanowire (GNWs) was developed for the detection of a model protein. Human immunoglobulin G (H-IgG) was selected as model analyte because of its physical, chemical, and biological features like many other biomarkers. GNWs were obtained through electrochemical template deposition into nanoporous polycarbonate membrane. To achieve the best morphology, various deposition conditions have been analyzed such as deposition time and Au precursor concentration in the deposition bath. The best deposition conditions were selected, ensuring the formation of mechanically stable nanowires. To detect proteins, a sandwich configuration was assembled on the surface of the electrode through several incubation steps. The sandwich configuration consists of a) a primary antibody covalently attached on the electrode surface, b) the antigen to be detected (H-IgG) that is selectively bound by the primary antibody, and c) a secondary labelled antibody. The immunosensor is electrochemically active thanks to the presence of gold nanoparticles (GNPs) tagging the secondary antibody. Given that GNPs catalyze hydrogen evolution reaction, the electrode has been used to measure the current density of the hydrogen evolution, which is indirectly related to the concentration of H-IgG antigens. In this way the calibration curve was constructed obtaining linear range of 1-1000 ng mL-1 with a high sensitivity

Electrochemical Quantification of H2O2 Released by Airway Cells Growing in Different Culture Media

Quantification of oxidative stress is a challenging task that can help in monitoring chronic inflammatory respiratory airway diseases. Different studies can be found in the literature regarding the development of electrochemical sensors for H2O2 in cell culture medium to quantify oxidative stress. However, there are very limited data regarding the impact of the cell culture medium on the electrochemical quantification of H2O2. In this work, we studied the effect of different media (RPMI, MEM, DMEM, Ham's F12 and BEGM/DMEM) on the electrochemical quantification of H2O2. The used electrode is based on reduced graphene oxide (rGO) and gold nanoparticles (AuNPs) and was obtained by co-electrodeposition. To reduce the electrode fouling by the medium, the effect of dilution was investigated using diluted (50% v/v in PBS) and undiluted media. With the same aim, two electrochemical techniques were employed, chronoamperometry (CH) and linear scan voltammetry (LSV). The influence of different interfering species and the effect of the operating temperature of 37 degrees C were also studied in order to simulate the operation of the sensor in the culture plate. The LSV technique made the sensor adaptable to undiluted media because the test time is short, compared with the CH technique, reducing the electrode fouling. The long-term stability of the sensors was also evaluated by testing different storage conditions. By storing the electrode at 4 degrees C, the sensor performance was not reduced for up to 21 days. The sensors were validated measuring H2O2 released by two different human bronchial epithelial cell lines (A549, 16HBE) and human primary bronchial epithelial cells (PBEC) grown in RPMI, MEM and BEGM/DMEM media. To confirm the results obtained with the sensor, the release of reactive oxygen species was also evaluated with a standard flow cytometry technique. The results obtained with the two techniques were very similar. Thus, the LSV technique permits using the proposed sensor for an effective oxidative stress quantification in different culture media and without dilution

Evaluation of the Purity of Magnesium Hydroxide Recovered from Saltwork Bitterns

Magnesium has been listed among the 30 critical raw materials by the European Union. In recent years, many green and sustainable alternative Mg2+ sources have been sought to satisfy the EU’s demand and to avoid mineral ore consumption. In this context, saltwork bitterns, the by-products of solar sea salt production, have attracted much attention thanks to their high Mg2+ concentrations (up to 80 g/L) and low Ca2+ and bicarbonate contents (95% (w/w). This work presents a comprehensive experimental effort of reactive precipitation tests with NaOH solutions at stoichiometric and over-stoichiometric concentrations to: (i) assess the technical feasibility of Mg2+ recovery from real bitterns collected in saltworks of the Trapani district (Italy) and, (ii) for the first time, conduct an extensive purity investigation of the precipitated magnesium hydroxide powders as brucite. This experimental investigation demonstrates the possibility of extracting highly valuable compounds from saltwork bittern waste, embracing the water valorization and resource recovery approach

Clustering of risk factors in hypertensive insulin-dependent diabetics with high sodium-lithium countertransport

Clustering of risk factors in hypertensive insulin-dependent diabetics with high sodium-lithium countertransport. Diabetic nephropathy is more common in patients with a positive family history of hypertension and with elevated red blood cell sodium-lithium countertransport, a marker of risk for essential hypertension. To evaluate whether there is a relationship between this cation transport system and indicators of risk of renal and cardiovascular complications in diabetic patients before the development of clinical proteinuria, we studied 31 type 1 (insulin-dependent) diabetic patients with arterial hypertension, without clinical proteinuria and 12 normotensive normoalbuminuric diabetic patients. Sodium-lithium countertransport activity was significantly higher in hypertensive patients (0.43 ± 0.03 mmol/1 RBC x hr) than in normotensive patients (0.23 ± 0.03; P < 0.001). To better explore the nature of the association between this transport system and arterial hypertension, hypertensive patients were divided in two groups, with high (>0.41 mmol/1 RBC x hr) or normal (<0.41) sodium-lithium countertransport activity. The two groups of hypertensive diabetics were similar in age, sex, body mass index and blood pressure levels. Hypertensive patients with elevated rates of sodium-lithium counter-transport compared with those with normal sodium-lithium counter-transport activity showed elevated glomerular filtration rate (130 ± 4 ml/min/1.73 m2 vs. 122 ± 3; P < 0.05), albumin excretion rate (median 26 /Lcg/min vs. 11; P < 0.001), higher fractional proximal sodium reabsorption (74 ± 1.2% vs. 71.6 ± 0.9; P < 0.01), exchangeable sodium pool (2937 ± 62 mmol/1.73 m2 vs. 2767 ± 56; P < 0.01), larger kidney volume (317 ± 7 ml/1.73 m2 vs. 270 ± 8; P < 0.05) and left ventricular mass index (122 ± 4 g/m2 vs. 107 ± 5; P < 0.05). Hypertensive patients with normal sodium-lithium countertransport activity had renal parameters similar to normotensive diabetic patients, except higher left ventricular mass index and kidney volume. Hypertensive diabetic patients with elevated sodium-lithium countertransport activity also had higher levels of plasma triglycerides, lower plasma HDL-cholesterol and impaired insulin sensitivity (assessed by euglyce-mic insulin-glucose clamp) compared with the other two groups. In conclusion, renal, cardiac and metabolic abnormalities are prominent in hypertensive type 1 (insulin-dependent) diabetic patients with higher sodium-lithium countertransport

In-silico guided chemical exploration of KDM4A fragments hits

Abstract Background Lysine demethylase enzymes (KDMs) are an emerging class of therapeutic targets, that catalyse the removal of methyl marks from histone lysine residues regulating chromatin structure and gene expression. KDM4A isoform plays an important role in the epigenetic dysregulation in various cancers and is linked to aggressive disease and poor clinical outcomes. Despite several efforts, the KDM4 family lacks successful specific molecular inhibitors. Results Herein, starting from a structure-based fragments virtual screening campaign we developed a synergic framework as a guide to rationally design efficient KDM4A inhibitors. Commercial libraries were used to create a fragments collection and perform a virtual screening campaign combining docking and pharmacophore approaches. The most promising compounds were tested in-vitro by a Homogeneous Time-Resolved Fluorescence-based assay developed for identifying selective substrate-competitive inhibitors by means of inhibition of H3K9me3 peptide demethylation. 2-(methylcarbamoyl)isonicotinic acid was identified as a preliminary active fragment, displaying inhibition of KDM4A enzymatic activity. Its chemical exploration was deeply investigated by computational and experimental approaches which allowed a rational fragment growing process. The in-silico studies guided the development of derivatives designed as expansion of the primary fragment hit and provided further knowledge on the structure–activity relationship. Conclusions Our study describes useful insights into key ligand-KDM4A protein interaction and provides structural features for the development of successful selective KDM4A inhibitors

Caspase-8 activation by cigarette smoke induces pro-inflammatory cell death of human macrophages exposed to lipopolysaccharide

Abstract Cigarette smoking impairs the lung innate immune response making smokers more susceptible to infections and severe symptoms. Dysregulation of cell death is emerging as a key player in chronic inflammatory conditions. We have recently reported that short exposure of human monocyte-derived macrophages (hMDMs) to cigarette smoke extract (CSE) altered the TLR4-dependent response to lipopolysaccharide (LPS). CSE caused inhibition of the MyD88-dependent inflammatory response and activation of TRIF/caspase-8/caspase-1 pathway leading to Gasdermin D (GSDMD) cleavage and increased cell permeability. Herein, we tested the hypothesis that activation of caspase-8 by CSE increased pro-inflammatory cell death of LPS-stimulated macrophages. To this purpose, we measured apoptotic and pyroptotic markers as well as the expression/release of pro-inflammatory mediators in hMDMs exposed to LPS and CSE, alone or in combination, for 6 and 24 h. We show that LPS/CSE-treated hMDMs, but not cells treated with CSE or LPS alone, underwent lytic cell death (LDH release) and displayed apoptotic features (activation of caspase-8 and -3/7, nuclear condensation, and mitochondrial membrane depolarization). Moreover, the negative regulator of caspase-8, coded by CFLAR gene, was downregulated by CSE. Activation of caspase-3 led to Gasdermin E (GSDME) cleavage. Notably, lytic cell death caused the release of the damage-associated molecular patterns (DAMPs) heat shock protein-60 (HSP60) and S100A8/A9. This was accompanied by an impaired inflammatory response resulting in inhibited and delayed release of IL6 and TNF. Of note, increased cleaved caspase-3, higher levels of GSDME and altered expression of cell death-associated genes were found in alveolar macrophages of smoker subjects compared to non-smoking controls. Overall, our findings show that CSE sensitizes human macrophages to cell death by promoting pyroptotic and apoptotic pathways upon encountering LPS. We propose that while the delayed inflammatory response may result in ineffective defenses against infections, the observed cell death associated with DAMP release may contribute to establish chronic inflammation. CS exposure sensitizes human macrophages to pro-inflammatory cell death. Upon exposure to LPS, CS inhibits the TLR4/MyD88 inflammatory response, downregulating the pro-inflammatory genes TNF and IL6 and the anti-apoptotic gene CFLAR, known to counteract caspase-8 activity. CS enhances caspase-8 activation through TLR4/TRIF, with a partial involvement of RIPK1, resulting on the activation of caspase-1/GSDMD axis leading to increased cell permeability and DAMP release through gasdermin pores [19]. At later timepoints caspase-3 becomes strongly activated by caspase-8 triggering apoptotic events which are associated with mitochondrial membrane depolarization, gasdermin E cleavage and secondary necrosis with consequent massive DAMP release