Salt Metabolism Research Articles

The relationship between gallstone disease and the genetic variants of the sterol transporter adenosine triphosphate–binding cassette G8 and G5 in Egyptians

BackgroundGallstones are abnormal lumps in the gallbladder or biliary tract due impaired cholesterol, bilirubin, or bile salt metabolism. The Adenosine triphosphate binding cassette transporter genes G5 and G8 (ABCG5, ABCG8) are two half transporters which work together as a heterodimer to regulate cholesterol levels in bile, and any alterations in their function can contribute to gallstone formation. The primary objective of this study was to evaluate the association between three specific polymorphisms—ABCG5 i7892T > C, ABCG5 Q604E, and ABCG8 D19H—and the risk of gallstone disease (GSD) in Egyptian females. These polymorphisms result from nucleotide substitutions in the gene sequences, which affect the transporter’s ability to efficiently regulate cholesterol secretion into the bile. This alteration can lead to cholesterol supersaturation in the bile, a key factor in the development of cholesterol gallstones. Additionally, the study aimed to examine the impact of these genetic variations on serum lipid profile to understand their role in modulating biochemical markers associated with GSD. Furthermore, the study sought to investigate haplotype patterns and explore their combined effects on disease susceptibility, providing deeper insight into the genetic factors that contribute to the development of GSD.MethodsThe study included 100 female patients diagnosed with gallstones and 100 healthy controls. Genotyping of single nucleotide polymorphisms (SNPs) was performed using allelic discrimination pre-designed TaqMan polymerase chain reaction method. Various laboratory investigations were measured using enzymatic colorimetric methods, and hematology analyzer was used for the whole blood count test.ResultsBetween patients with gallstone disease and healthy controls, there were statistically significant differences in the distribution of these genes polymorphisms. Q604E CC genotype (OR = 15.2; P = 0.004) and C allele (OR = 2; P = 0.007) in ABCG5 (rs6720173) as well as D19H GC genotype (OR = 2.9; P = 0.002) and C allele (OR = 2; P = 0.004) in ABCG8 (rs11887534) were significantly more frequent in gallstone patients. The CCC haplotype is a statistically significant predictor of GSD.ConclusionsThis study suggests that ABCG8 D19H (G/C) and ABCG5 Q604E (C/C) genotypes may play a significant role in GSD susceptibility among Egyptian females.Graphical abstract

An In Silico Investigation of the Pathogenic G151R G Protein-Gated Inwardly Rectifying K+ Channel 4 Variant to Identify Small Molecule Modulators

Primary aldosteronism is characterised by the excessive production of aldosterone, which is a key regulator of salt metabolism, and is the most common cause of secondary hypertension. Studies have investigated the association between primary aldosteronism and genetic alterations, with pathogenic mutations being identified. This includes a glycine-to-arginine substitution at position 151 (G151R) of the G protein-activated inward rectifier potassium (K+) channel 4 (GIRK4), which is encoded by the KCNJ5 gene. Mutations in GIRK4 have been found to reduce the selectivity for K+ ions, resulting in membrane depolarisation, the activation of voltage-gated Ca2+ channels, and an increase in aldosterone secretion. As a result, there is an interest in identifying and exploring the mechanisms of action of small molecule modulators of wildtype (WT) and mutant channels. In order to investigate the potential modulation of homotetrameric GIRK4WT and GIRK4G151R channels, homology models were generated. Molecular dynamics (MD) simulations were performed, followed by a cluster analysis to extract starting structures for molecular docking. The central cavity has been previously identified as a binding site for small molecules, including natural compounds. The OliveNetTM database, which consists of over 600 compounds from Olea europaea, was subsequently screened against the central cavity. The binding affinities and interactions of the docked ligands against the GIRK4WT and GIRK4G151R channels were then examined. Based on the results, luteolin-7-O-rutinoside, pheophorbide a, and corosolic acid were identified as potential lead compounds. The modulatory activity of olive-derived compounds against the WT and mutated forms of the GIRK4 channel can be evaluated further in vitro.

ANP Improves The Water And Salt Metabolism During Cooling Treatment For Exertional Heat Stroke

ANP Improves The Water And Salt Metabolism During Cooling Treatment For Exertional Heat Stroke

Unveiling the Impact of Lactic Acid Bacteria on Blood Lipid Regulation for Cardiovascular Health

Lactic acid bacteria (LAB) are a group of microorganisms which are beneficial and well-characterized with respect to the flavor and texture of food products via fermentation. The accumulated literature has suggested that dietary intake of fermented foods rich in LAB is related to different health-promoting benefits; however, in recent years, emerging evidence suggests a contribution of LAB to blood lipid regulation and cardiovascular health via certain mechanisms. Different potential mechanisms for the lipid regulatory effects of LAB may include the interaction of hydroxymethylglutaryl-CoA (HMG-CoA) reductase and bile salt hydrolase activity and bile salt metabolism; gut microbiome modulation; and regulation of mRNA expression of genes related to fat metabolism in animal models and human studies. This review comprehensively aims to answer whether/how LAB influence blood lipids in both animal models and human studies while also uncovering the underlying mechanisms linking LAB to lipid metabolism.

Spot urinary sodium in CKD patients: correlation with 24h-excretion and evaluation of commonly used prediction equations

BackgroundSalt intake in CKD patients can affect cardiovascular risk and kidney disease progression. Twenty-four hour (24h) urine collections are often used to investigate salt metabolism but are cumbersome to perform. We assessed urinary sodium (U-Na) concentration in spot urine samples and investigated the correlation with 24h U-Na excretion and concentration in CKD patients under nephrological care. Further, we studied the role of CKD stage and diuretics and evaluated the performance of commonly used formulas for the prediction of 24h U-Na excretion from spot urine samples.MethodsOne hundred eight patients of the German Chronic Kidney Disease (GCKD) study were included. Each participant collected a 24h urine and two spot urine samples within the same period. The first spot urine sample (AM) was part of the second morning urine. The second urine sample was collected before dinner (PM). Patients were advised to take their medication as usual without changing dietary habits. U-Na concentrations in the two spot urine samples and their average ((AM + PM)/2) were correlated with U-Na concentration and total Na excretion in the 24h urine collections. Correlations were subsequently studied after stratification by CKD stage and diuretic intake. The usefulness of three commonly applied equations to estimate 24h U-Na excretion from spot urine samples (Kawasaki, Tanaka and Intersalt) was determined using Bland–Altman plots, analyses of sensitivity, specificity, as well as positive (PPV) and negative predictive values (NPV).ResultsParticipants (42 women, 66 men) were on average (± SD) 62.2 (± 11.9) years old, with a mean serum creatinine of 1.6 (± 0.5) mg/dl. 95% had arterial hypertension, 37% diabetes mellitus and 55% were on diuretics. The best correlation with 24h U-Na total excretion was found for the PM spot U-Na sample. We also found strong correlations when comparing spot and 24h urine U-Na concentration. Correction of spot U-Na for U-creatinine did not improve strength of correlations. Neither CKD stage, nor intake of diuretics had significant impact on these correlations. All examined formulas revealed a significant mean bias. The lowest mean bias and the strongest correlation between estimated and measured U-Na excretion in 24h were obtained using the Tanaka-formula. Also, application of the Tanaka-formula with PM U-Na provided best sensitivity, specificity, PPV and NPV to estimate U-Na excretion > 4g/d corresponding to a salt consumption > 10g/d.ConclusionU-Na concentration of spot urine samples correlated with 24h U-Na excretion especially when PM spot U-Na was used. However, correlation coefficients were relatively low. Neither CKD stage nor intake of diuretics appeared to have an influence on these correlations. There was a significant bias for all tested formulas with the Tanaka-formula providing the strongest correlation with measured 24h U-Na excretion. In summary, using spot urine samples together with the Tanaka-formula in epidemiological studies appears feasible to determine associations between approximate salt intake and outcomes in CKD patients. However, the usefulness of spot-urine samples to guide and monitor salt consumption in individual patients remains limited.

Can the genetic background modulate the effects of feed additives? Answers from gut microbiome and transcriptome interactions in farmed gilthead sea bream (Sparus aurata) fed with a mix of phytogenics, organic acids or probiotics

The synergies between selective breeding and feed additives remain under-explored in farmed fish, despite their sustainability. Reference (REF) and selected gilthead sea bream for growth (GS) were fed with the control (CTRL) diet during 14 days. CTRL diet was oil-coated with three functional additives (PHY: phytogenic based on garlic and medium chain fatty acid; OA: organic acid mixture with a 70% of butyric acid sodium salt; PROB: probiotic based on Bacillus subtilis, pumillus and licheniformes species). These experimental diets were then sequentially administered at high (PHY/OA = 7.5 g/kg, PROB = 2 × 1011 CFU/kg; 2 weeks) and low (PHY = 5 g/kg, OA = 3 g/kg, PROB = 4 × 1010 CFU/kg; 10 weeks) additive doses. The capacity of a given genotype and additive to modify the fish growth performance, gut health and the host interaction with its anterior intestine (AI) microbiota was evaluated as a whole population or individually (9 fish/diet/genetics). GS fish showed a better growth and feed conversion ratio, linked to a reduced individual variability of gut microbial composition. The PHY additive had a major impact upon the intestinal transcriptome of GS-PHY fish, with the up-regulation of markers of epithelial integrity, sphingolipid and cholesterol/bile salt metabolism. With the OA additive, impaired growth performance, reduced AI goblet cell area and enhanced AI granulocyte infiltration were concomitant with a down-regulation of neutrophil degranulation markers associated with a decrease of pathogenic genera (Staphylococcus/Streptococcus/Neisseria), and an over-representation of acetone/butanol/ethanol fermentation and vitamin K biosynthesis inferred pathways. Bacillus establishment and lack of AI inflammation were parallel in PROB fish of both genetic backgrounds. However, GS fish grew and utilized feed better with the additive, whereas a worsening appeared in REF fish. This amelioration was related with a higher abundance of the nitrate-reducer Kocuria, an up-regulation of markers of epithelial cell maintenance and proliferation, and a down-regulation of microbiota-correlated protein synthesis and ubiquitination markers, supporting a reduced epithelial turnover and improved intestinal barrier function. Overall, the success of nutritional innovations in gilthead sea bream is largely dependent on the host genome predisposition, but also on the intestinal microbiota according to the hologenome theory.

Construction and validation of a joint diagnosis model based on random forest and artificial intelligence network for hepatitis B-related hepatocellular carcinoma.

Hepatitis B virus (HBV) is the dominant pathogenic factor of hepatocellular carcinoma (HCC) in Asia and Africa. Early identification and clinical diagnosis are crucial for HBV-related HCC. Random forest (RF) and artificial neural network (ANN) were an innovative and highly effective supervised machine learning (ML) algorithm for the early diagnosis and screening of HBV-related HCC. This study aims to identify significant biomarkers and develop a novel genetic model for the efficient diagnosis of HBV-related HCC. Gene Expression Omnibus (GEO) Series (GSE)19665, GSE55092, and GSE121248 were used to identify significant differentially expressed genes (DEGs). The enrichment analysis was performed on Metascape online tool. The RF algorithm and ANN were used to select the potential predictive gene panels and construct an HBV-related HCC diagnostic model. Subsequently, GSE17548, GSE104310, GSE44074, and GSE136247 were used to test the accuracy of the ANN model. Finally, the CIBERSORT algorithm was used to assess the abundance of immune infiltrates in all samples. First, 116 genes were identified as DEGs, and the DEGs were particularly enriched in cellular hormone metabolic process, monocarboxylic acid metabolic process, NABA extracellular matrix (ECM) AFFILIATED steroid metabolic process and metabolism of bile acid and bile salt. DNA topoisomerase II alpha (TOP2A), C-type lectin domain family 1 member B (CLEC1B), BUB1 mitotic checkpoint serine/threonine kinase B (BUB1B), ficolin 2 (FCN2), C-X-C motif chemokine ligand 14 (CXCL14), cyclase associated actin cytoskeleton regulatory protein 2 (CAP2), ficolin 3 (FCN3), kynurenine 3-monooxygenase (KMO) and cadherin related family member 2 (CDHR2) were available to develop an HBV-related HCC diagnostic model. After validation, the diagnostic model showed high sensitivity (88.5%, 90%, 88.5%, 76.5%) and specificity (100%, 81.8%, 89.5%, 72.2%), and the areas under the receiver operating characteristic (ROC) curves showed excellent efficiency (1, 0.927, 0.921, 0.833). Finally, the percentage of infiltrating immune cell types [B cells naïve, B cells memory, plasma cells, T cells CD8, T cells CD4 memory resting, T cells regulatory (Tregs), T cells gamma delta, natural killer (NK) cells resting, NK cells activated, Macrophages M0, Dendritic cells activated, Mast cells activated] for hepatitis B-related HCC were significantly different from that of non-cancerous liver tissue with HBV. A novel early diagnostic model of HBV-related HCC was established, and the model showed better efficiency in distinguishing HBV-related HCC from other non-cancerous with HBV individuals.

Pathological evaluation of the effects of the drug clozalbent f on tissues and cytostructure of the trematode <i>Fasciola gigantica</i> (Cobbold, 1855)

The aim of the study was a pathomorphological evaluation of the effect of the new drug Clozalbent F (Clozalbent F) on populations of the trematode Fasciola gigantica (Cobbold, 1855) with monoinvasion of fasciolеsis in cows. In the experiment, on days 7-10 after a single oral application to cows spontaneously infected with trematodes Fasciola gigantica, the effects of the drug Clozalbent F (Clozalbent F) at a dose of 1.25 g/10 kg of live weight appear in the form of irreversible pathomorphological changes in organs and tissues marit and premarital. In the body of trematodes, carbohydrate, protein, water and salt metabolism is disturbed, toxins are formed, and hydropic dystrophy of tissues increases with large amounts of water in them. Hydration of F. gigantica cells and tissues causes them to swell, then decay and necrosis. Under the influence of Closalbent F, vacuoles are formed in the tegument of trematodes in the intercellular spaces filled with turbid moisture. The drug enters the body of F. gigantica through the neo-dermis, causing irreversible path changes (destruction, disorganisation, decomposition, autolysis, protein, carbohydrate, hydropic dystrophy, necrobiosis and necrosis) of cells and tissues in mature trematodes. On the 7-10th day of the experiment, after the action of the Clozalbent F at a dose of 1,25 g/10 kg of live weight, F. gigantica marita showed complete disintegration of tegument, suckers and genitalia cells, which was confirmed by histological methods (TS, BFS, SA, CHIC) and testifying to its high efficiency and biosafety fasciolеsis in cow.

Physicochemical, microbiological and metabolomics changes in yogurt supplemented with lactosucrose

Lactosucrose (LS) is a known prebiotic that has gained recognition for its low caloric content and various health benefits. However, its potential in food applications remains largely unexplored. In this study the effects of adding LS to milk at concentrations (0 %, 2 %, 5 % and 8 % w/v) for yogurt production, and the relevant changes in yogurt texture, microbial composition and metabolomics were investigated. Our findings revealed that LS played a role in promoting the formation of a structured gel during fermentation, resulting in increased elasticity and viscosity while reducing fluidity. Additionally incorporating high doses of LS into yogurt led to reduced post-acidification, enhanced survival of starter bacteria, improved water retention capacity and overall texture throughout a refrigerated storage period of 21 days. Notably higher concentrations of LS (8 % w/v) exhibited effects on enhancing yogurt quality. Furthermore, untargeted metabolomics analysis using UPLC Q TOF MS/MS revealed 45 differentially expressed metabolites, including up-regulated L-arginine, L-proline and L-glutamic acid along with the down-regulated glutathione, L-tyrosine, L-phenylalanyl and L-proline. These differential metabolites were primarily associated with amino acid metabolism such as thiamine metabolism, nicotinic acid salt and nicotinamide metabolism, and pyrimidine metabolism. As a result, the inclusion of LS in yogurt had an impact on the production of various beneficial metabolites in yogurt, highlighting the importance of combining prebiotic LS with probiotics to obtain desired physiological benefits of yogurt.

Genome analysis of haloalkaline isolates from the soda saline crater lake of Isabel Island; comparative genomics and potential metabolic analysis within the genus Halomonas

BackgroundIsabel Island is a Mexican volcanic island primarily composed of basaltic stones. It features a maar known as Laguna Fragatas, which is classified as a meromictic thalassohaline lake. The constant deposition of guano in this maar results in increased levels of phosphorus, nitrogen, and carbon. The aim of this study was to utilize high-quality genomes from the genus Halomonas found in specialized databases as a reference for genome mining of moderately halophilic bacteria isolated from Laguna Fragatas. This research involved genomic comparisons employing phylogenetic, pangenomic, and metabolic-inference approaches.ResultsThe Halomonas genus exhibited a large open pangenome, but several genes associated with salt metabolism and homeostatic regulation (ectABC and betABC), nitrogen intake through nitrate and nitrite transporters (nasA, and narGI), and phosphorus uptake (pstABCS) were shared among the Halomonas isolates.ConclusionsThe isolated bacteria demonstrate consistent adaptation to high salt concentrations, and their nitrogen and phosphorus uptake mechanisms are highly optimized. This optimization is expected in an extremophile environment characterized by minimal disturbances or abrupt seasonal variations. The primary significance of this study lies in the dearth of genomic information available for this saline and low-disturbance environment. This makes it important for ecosystem conservation and enabling an exploration of its biotechnological potential. Additionally, the study presents the first two draft genomes of H. janggokensis.

Metabolic interpretation of NaCl stress-induced lipid accumulation in microalgae for promising biodiesel production with saline wastewater

The rising use of saline wastewater for biodiesel production through oleaginous microalgae demands fundamental studies on the metabolic response of microalgal cells to salt stress. Thus, we investigated how salt stress promotes lipid accumulation inside Botryococcus sp. NJD-1 by tracking critical metabolites and proteases as well as analyzing the cellular redox balance. The results showed that the lipid content of microalgal cells followed a negative binomial correlation with intracellular reactive oxygen species (ROS), while the neutral lipid content increased linearly with salinity in range of 0–20 g/L NaCl. The optimal lipid production was achieved as 54.5 % and 110.5 mg/L d-1 at 10 g/L NaCl. Further comparing the metabolomics and proteomics data at 0 and 10 g/L NaCl, the adenosine triphosphate (ATP) and nicotinamide adenine nucleotide phosphate (NADPH) were found significantly increased to quench the Na+-induced ROS. The created NADPH and ATP accord the enhanced activity of the upper Pyruvate-Malate cycle and tricarboxylic acid cycle (TCA) cycle. Therefore, ROS detoxification is proposed to be the essential mechanism for microalgal lipid accumulation under salt stress. The cellular triacylglycerol (TAG) synthesis triggered by salinity is mainly mediated by an acetyl-CoA-dependent Kennedy pathway. These findings will deepen our understanding of the nexus of ambient salt, intracellular ROS and lipid metabolism for microalgae and advance the exploit of saline wastewater for microalgal lipid production.

Correlation analysis and clinical significance of Musculoskeletal Ultrasound semi-quantitative grading with bone salt metabolism, Rheumatoid factor and Erythrocyte Sedimentation rate in patients with Rheumatoid Arthritis.

To determine the correlation and clinical significance of musculoskeletal ultrasound semi-quantitative grading with bone salt metabolism, rheumatoid factor and erythrocyte sedimentation rate (ESR) in patients with rheumatoid arthritis. This is a clinical comparative study. A total of 240 patients with rheumatoid arthritis admitted to Baoding NO.1 Central Hospital were selected according to the DAS28 score of rheumatoid arthritis, and were divided into four groups, with 60 cases in each group from May 2020 to May 2022. The differences and correlation of musculoskeletal ultrasound semi-quantitative grading, bone metabolism indicators, erythrocyte sedimentation rate and rheumatoid factor among the four groups were statistically analyzed. The scores of bone erosion, synovial hyperplasia, joint effusion and intrasynovial blood flow in Group-H were significantly higher than those in Group-R, L and M, with statistically significant differences(p=0.00). The procollagen Type-1 N-terminal propeptide(P1NP), bone-specific alkaline phosphatase(BALP) and osteoprotegerin(OPG) in Group-H were significantly lower than those in Group-R, L and M, with statistically significant differences(p=0.00); The tartrate-resistant acid phosphatase(TRAC) in Group-H was significantly higher than that in Group-R, L and M, with a statistically significant difference(p=0.00). The levels of RF and ESR in Group-H were significantly higher than those in Group-R, L and M, with statistically significant differences(p=0.00). Musculoskeletal ultrasound semi-quantitative grading is correlated with the level of bone salt metabolism, rheumatoid factor and ESR in patients with rheumatoid arthritis. It can be combined with laboratory examination to objectively judge the severity of the course of rheumatoid arthritis.

Clinical efficacy of Thalidomide combined with PAD Regimen in patients with multiple Myeloma.

To evaluate the clinical efficacy of thalidomide combined with PAD regimen in patients with multiple myeloma (MM). It was a Clinical comparative study. A total of 120 patients with MM were admitted at Beijing Aerospace General Hospital from September 2020 to June 2022 randomly divided into two groups, with 60 patients in each group. The study group was treated with thalidomide combined with a PAD regimen (bortezomib, doxorubicin and dexamethasone), while the control group with a PAD regimen alone. After treatment, the therapeutic effect, adverse drug reactions, bone metabolic markers such as serum alkaline phosphatase (ALP) and osteocalcin (OCN) before and after treatment, as well as T-lymphocyte subsets CD3+, CD4+, CD8+ and CD4+/CD8+ levels before and after treatment were compared and analyzed between the two groups. The total efficacy in the study group was 90%, which was significantly higher than 70% in the control group (p= 0.00). The incidence of adverse drug reactions was 40% in the study group and 38% in the control group, without statistically significant difference (p= 0.85). After treatment, ALP and OCN levels in the study group were significantly higher than those in the control group (ALP, p= 0.01; OCN, p= 0.00), and CD3+, CD4+ and CD4+/CD8+ in the study group also increased significantly compared with those in the control group (CD3+, p= 0.02; CD4+, p= 0.00; CD4+/CD8+, p= 0.00). Thalidomide combined with a PAD regimen is definitely effective in patients with MM, it can obviously improve immune function and bone salt metabolism, with no increase in adverse reactions but high safety and effectiveness.

The Role of Palmitic Acid in the Co-Toxicity of Bacterial Metabolites to Endothelial Cells.

Metabolic endotoxemia most often results from obesity and is accompanied by an increase in the permeability of the intestinal epithelial barrier, allowing co-absorption of bacterial metabolites and diet-derived fatty acids into the bloodstream. A high-fat diet (HFD) leading to obesity is a significant extrinsic factor in developing vascular atherosclerosis. In this study, we evaluated the effects of palmitic acid (PA) as a representative of long-chain saturated fatty acids (LCSFA) commonly present in HFDs, along with endotoxin (LPS; lipopolysaccharide) and uremic toxin indoxyl sulfate (IS), on human vascular endothelial cells (HUVECs). HUVECs viability was measured based on tetrazolium salt metabolism, and cell morphology was assessed with fluorescein-phalloidin staining of cells' actin cytoskeleton. The effects of simultaneous treatment of endothelial cells with PA, LPS, and IS on nitro-oxidative stress in vascular cells were evaluated quantitatively with fluorescent probes. The expression of vascular cell adhesion molecule VCAM-1, E-selectin, and occludin, an essential tight junction protein, in HUVECs treated with these metabolites was evaluated in Western blot. PA, combined with LPS and IS, did not influence HUVECs viability but induced stress on actin fibers and focal adhesion complexes. Moreover, PA combined with LPS significantly enhanced reactive oxygen species (ROS) production in HUVECs but decreased nitric oxide (NO) generation. PA also considerably increased the expression of VCAM-1 and E-selectin in HUVECs treated with LPS or IS but decreased occludin expression. Palmitic acid enhances the toxic effect of metabolic endotoxemia on the vascular endothelium.

Phytocrystallization of silver nanoparticles using Cassia alata flower extract for effective control of fungal skin pathogens

Abstract A feasible alternative to classic chemical synthesis, the phyto-mediated production of silver nanoparticles (AgNPs) utilizing aqueous flower petal extract of Cassia alata as a reducing agent is reported for the first time. Characterization of synthesized AgNPs was carried out using various techniques viz., ultraviolet-visible spectroscopy (UV-Vis), X-ray powder diffraction (XRD), high-resolution transmission electron microscope (HRTEM), Fourier transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM) with energy dispersive X-ray analysis (EDX). The results of the FTIR research conducted in this study show different bond stretches with varying durations, which can be seen at various faraway points. AgNPs are mainly spherical and vary in size from 20 to 100 nm, according to TEM images. The highest X-ray energy surge, at 3 keV, is visible in the EDX spectrum. The XRD pattern showed that four diffraction peaks could be assigned to the 111, 200, 220, and 311 planes of the face-centered cubic crystalline silver, respectively, at 32.05, 46.27, 55.25, and 57.39°. Optimization of production parameters including pH, metal ion concentration, and substrate concentrations were studied. In addition, the bioactivity was evaluated against Trichophyton rubrum, Aspergillus fumigatus, Candida albicans, Epidermophyton floccosum, and Mucor sp. using the agar diffusion method. Furthermore, their antioxidant properties were assessed using 2,2-diphenyl-1-picryl-hydrazyl-hydrate assay and ferric ion reducing antioxidant power tests. MTT assay was performed using human fibroblast cell line (L929) to determine the cell viability and cytotoxicity through increased metabolism of the tetrazolium salt.

Antioxidant Supplementation Alleviates Mercury-Induced Cytotoxicity and Restores the Implantation-Related Functions of Primary Human Endometrial Cells.

Mercury (Hg) cytotoxicity, which is largely mediated through oxidative stress (OS), can be relieved with antioxidants. Thus, we aimed to study the effects of Hg alone or in combination with 5 nM N-Acetyl-L-cysteine (NAC) on the primary endometrial cells' viability and function. Primary human endometrial epithelial cells (hEnEC) and stromal cells (hEnSC) were isolated from 44 endometrial biopsies obtained from healthy donors. The viability of treated endometrial and JEG-3 trophoblast cells was evaluated via tetrazolium salt metabolism. Cell death and DNA integrity were quantified following annexin V and TUNEL staining, while the reactive oxygen species (ROS) levels were quantified following DCFDA staining. Decidualization was assessed through secreted prolactin and the insulin-like growth factor-binding protein 1 (IGFBP1) in cultured media. JEG-3 spheroids were co-cultured with the hEnEC and decidual hEnSC to assess trophoblast adhesion and outgrowth on the decidual stroma, respectively. Hg compromised cell viability and amplified ROS production in trophoblast and endometrial cells and exacerbated cell death and DNA damage in trophoblast cells, impairing trophoblast adhesion and outgrowth. NAC supplementation significantly restored cell viability, trophoblast adhesion, and outgrowth. As these effects were accompanied by the significant decline in ROS production, our findings originally describe how implantation-related endometrial cell functions are restored in Hg-treated primary human endometrial co-cultures by antioxidant supplementation.

Modulation of gut microbiota and intestinal immune response in gilthead seabream (Sparus aurata) by dietary bile salt supplementation.

Given their role in lipid digestion, feed supplementation with bile salts could be an economic and sustainable solution to alterations in adiposity and intestinal inflammation generated by some strategies currently used in aquaculture. An important part of the metabolism of bile salts takes place in the intestine, where the microbiota transforms them into more toxic forms. Consequently, we aimed to evaluate the gut immune response and microbial populations in gilthead seabream (Sparus aurata) fed a diet supplemented with a blend of bile salts with proven background as a regulator of lipid metabolism and fat content. After the 90-day feeding trial, a differential modulation of the microbiota between the anterior and posterior intestine was observed. While in the anterior intestine the relative abundance of Desulfobacterota doubled, in the posterior intestine, the levels of Firmicutes increased and Proteobacteria, Actinobacteriota, and Campylobacterota were reduced when supplementing the diet with bile salts. Even so, only in the anterior intestine, there was a decrease in estimated richness (Chao1 and ACE indices) in presence of dietary bile salts. No significant differences were displayed in alpha (Shannon and Simpson indices) nor beta-diversity, showing that bile sales did not have a great impact on the intestinal microbiota. Regarding the gene expression profile in 2 h postprandial-fish, several changes were observed in the analyzed biomarkers of epithelial integrity, nutrient transport, mucus production, interleukins, cell markers, immunoglobulin production and pathogen recognition receptors. These results may indicate the development of an intestinal immune-protective status to tackle future threats. This work also suggests that this immune response is not only regulated by the presence of the dietary bile salts in the intestine, but also by the microbial populations that are in turn modulated by bile salts. After a fasting period of 2 days, the overall gene expression profile was stabilized with respect to fish fed the unsupplemented diet, indicating that the effect of bile salts was transient after short periods of fasting. On the balance, bile salts can be used as a dietary supplement to enhance S. aurata farming and production without compromising their intestinal health.

Regulation of lipid metabolism by gut microbiota in aquatic animals

AbstractLipids are an essential component of living beings and an important group of nutrients. As the gut microbiota plays important roles in the intestinal absorption and extraintestinal metabolism of dietary lipids, the current review addresses the recent progress regarding the interactions between the gut microbiota and lipid metabolism in aquatic animals, with a focus on fish. We discuss in detail how dietary lipid sources and content affect the composition of the gut microbiome and the mechanism by which the gut microbiota affects the lipid metabolism of the host. This interaction is largely mediated via microbial lipases, short‐chain fatty acids and the gut‐liver axis. The latter refers to the metabolism of biliary salts and acids, the regulation of their synthesis by gut microbes and their impact on the lipid metabolism. Finally, we briefly discuss how probiotic supplementation modulates the host's microbiome, and how probiotics have a beneficial effect on health and welfare of farmed aquatic animals. Although the influence of intestinal microbiota on lipid metabolism has been explored before, further research is needed to profoundly investigate the molecular mechanisms by which microbial metabolites (SCFAs and bile acids) induce lipid metabolism.

Hypercrystalluria as a Factor in the Development of Urine Stone Disease, Diagnosis and Directions of Treatment

Under the action of exogenous, androgenic, genetically determined factors, the metabolism of stone-forming salts of calcium, phosphorus, magnesium, oxalates, uric acid in the blood serum and their active excretion by the kidneys to the state of hypersaturation (oversaturation) is disturbed) urine is formed. When the level of crystallization inhibitors is disturbed, a saturated salt solution crystallizes with the formation of microliths. The formation of stones in the kidneys is possible only in the presence of «building material» – supersaturated saturated urine, therefore, hyperoxaluria is a pre-stone condition. Treatment measures should be aimed at correcting mineral metabolism in the body after establishing the type of hyperoxaluria using laboratory tests: salt transport, calcium load, dietary test – low-calcium diet, thiazide test and determination of the mineral composition of the removed (removed) stone. Genetically consequential conditions (10–15%) count about 30 varieties in which the main sign or symptom in the manifestation of the disease is urolithiasis. Unfortunately, congenital tubulopathies are not sufficiently studied, so the treatment is symptomatic, in some cases simultaneous kidney and liver transplantation options are possible. Clinically, 4 main forms of hypercrystalluria are distinguished: hypercalciuria, hyperoxaluria, hyperuricuria, phosphaturia and mixed forms of crystalluria. Acquired forms of hypercrystalluria, of which they are absorptive (type II intestinal hyperabsorption – absorptive hypercalciuria and absorptive hyperoxaluria), are of main clinical interest, which is characteristic of the course of calcium-oxalate urolithiasis. Metaphylaxis of calcium-oxalate urolithiasis is formed on the basis of these data.

CRISPRi-Mediated Gene Suppression Reveals Putative Reverse Transcriptase Gene PA0715 to Be a Global Regulator of Pseudomonas aeruginosa.

Pseudomonas aeruginosa is a common pathogen of infection in burn and trauma patients, and multi-drug resistant P. aeruginosa has become an increasingly important pathogen. Essential genes are key to the development of novel antibiotics. The PA0715 gene is a novel unidentified essential gene that has attracted our interest as a potential antibiotic target. Our study aims to determine the exact role of PA0715 in cell physiology and bacterial pathogenicity, providing important clues for antibiotic development. The shuttle vector pHERD20T containing an arabinose inducible promoter was used to construct the CRISPRi system. Alterations in cellular physiology and bacterial pathogenicity of P. aeruginosa PAO1 after PA0715 inhibition were characterized. High-throughput RNA-seq was performed to gain more insight into the mechanisms by which PA0715 regulates the vital activity of P. aeruginosa. We found that down-regulation of PA0715 significantly reduced PAO1 growth rate, motility and chemotaxis, antibiotic resistance, pyocyanin and biofilm production. In addition, PA0715 inhibition reduced the pathogenicity of PAO1 to the greater galleria mellonella larvae. Transcriptional profiling identified 1757 genes including those related to amino acid, carbohydrate, ketone body and organic salt metabolism, whose expression was directly or indirectly controlled by PA0715. Unexpectedly, genes involved in oxidative phosphorylation also varied with PA0715 levels, and these findings support a hitherto unrecognized critical role for PA0715 in oxidative respiration in P. aeruginosa. We identified PA0715 as a global regulator of the metabolic network that is indispensable for the survival and reproduction of P. aeruginosa. Our results provide a basis for future studies of potential antibiotic targets for P. aeruginosa and offer new ideas for P. aeruginosa infection control.