Hemolysis Of Erythrocytes Research Articles

Hemolysis by Saponin Is Accelerated at Hypertonic Conditions.

Saponins are a large group of organic amphiphilic substances (surfactants) mainly extracted from herbs with biological activity, considered as one of the main ingredients in numerous remedies used in traditional medicine since ancient times. Anti-inflammatory, antifungal, antibacterial, antiviral, antiparasitic, antitumor, antioxidant and many other properties have been confirmed for some. There is increasing interest in the elucidation of the mechanisms behind the effects of saponins on different cell types at the molecular level. In this regard, erythrocytes are a very welcome model, having very simple structures with no organelles. They react to changing external conditions and substances by changing shape or volume, with damage to their membrane ultimately leading to hemolysis. Hemolysis can be followed spectrophotometrically and provides valuable information about the type and extent of membrane damage. We investigated hemolysis of erythrocytes induced by various saponin concentrations in hypotonic, isotonic and hypertonic media using measurements of real time and end-point hemolysis. The osmotic pressure was adjusted by different concentrations of NaCl, manitol or a NaCl/manitol mixture. Unexpectedly, at a fixed saponin concentration, hemolysis was accelerated at hypertonic conditions, but was much faster in NaCl compared to mannitol solutions at the same osmotic pressure. These findings confirm the colloid-osmotic mechanism behind saponin hemolysis with pore formation with increasing size in the membrane.

Alpha hemolysin of E. coli induces hemolysis of human erythrocytes independently of toxin interaction with membrane proteins

Alpha hemolysin (HlyA) is a hemolytic and cytotoxic protein secreted by uropathogenic strains of E. coli. The role of glycophorins (GPs) as putative receptors for HlyA binding to red blood cells (RBCs) has been debated.Experiments using anti-GPA/GPB antibodies and a GPA-specific epitope nanobody to block HlyA-GP binding on hRBCs, showed no effect on hemolytic activity. Similarly, the hemolysis induced by HlyA remained unaffected when hRBCs from a GPAnull/GPBnull variant were used. Surface Plasmon Resonance experiments revealed similar values of the dissociation constant between GPA and either HlyA, ProHlyA (inactive protoxin), HlyAΔ914-936 (mutant of HlyA lacking the binding domain to GPA) or human serum albumin, indicating that the binding between the proteins and GPA is not specific.Although far Western blot followed by mass spectroscopy analyses suggested that HlyA interacts with Band 3 and spectrins, hemolytic experiments on spectrin-depleted hRBCs and spherocytes, indicated these proteins do not mediate the hemolytic process.Our results unequivocally demonstrate that neither glycophorins, nor Band 3 and spectrins mediate the cytotoxic activity of HlyA on hRBCs, thereby challenging the HlyA-receptor hypothesis. This finding holds significant relevance for the design of anti-toxin therapeutic strategies, particularly in light of the growing antibiotic resistance exhibited by bacteria.

Oxidative stress in animals exposed to cypermethrin: mechanisms of development and possible risks

The purpose of the study is to establish the mechanisms of development of oxidative stress during long-term low-dose exposure to cypermethrin on the animal organism and to substantiate possible risks. The work was carried out using Wistar rats, which were intragastrically injected daily with cypermethrin at a dose of 1/1000 LD50 for 60 and 120 days. At the same time, control animals received an appropriate volume of saline solution inside. When removing animals from the experiment, blood was taken to determine the number of red blood cells, the concentration of hemoglobin, glucose, lactic and uric acids, and total bilirubin. In the study of erythrocytes and liver, biochemical parameters characterizing carbohydrate and purine metabolism, processes of xenobiotics biotransformation, pro- and antioxidant processes were determined. Long-term lowdose exposure to cypermethrin for 60 days causes hematotoxic effects in animals, enhancement of xenobiotic biotransformation processes, and slight compensatory restructuring of metabolic processes in the liver. Exposure to 1/1000 LD50 of cypermethrin for 120 days, in addition to the above effects, causes the development of acidosis, enhances pro-oxidant processes, and suppresses the function of the antioxidant defense system and the pentose cycle. The decrease in the activity of the liver antioxidant system is due to glutathione deficiency due to the activation of xenobiotic biotransformation systems and suppression of the activity of glutathione reductase and glucose-6- phosphate dehydrogenase, which ultimately leads to the development of oxidative stress. Long-term exposure of animals to low doses of cypermethrin causes hemolysis of erythrocytes, lactic acidosis, activation of prooxidant systems, and inhibition of the antioxidant defense system. Deficiency of carbohydrates and suppression of the activity of the pentose cycle indicate a decrease in the functional activity of the liver and an increased risk of immunosuppression.

Investigative on the molecular mechanism of novel antioxidation peptides by 3D-QSAR, in vitro assay and MD simulations

Three novel antioxidant peptides MHW, YHW and FHW were designed by the analysis of 3D-QSAR. Two accurate CoMFA (Q2 = 0.886 and R2 = 0.988) and CoMSIA (Q2 = 0.737 and R2 = 0.955) models were established. The FTC assay showed that activities of the new peptides on lipid peroxidation is significantly better than that of the template molecule LHW. The results of erythrocyte hemolysis assay demonstrated that MHW, YHW and FHW significantly inhibited AAPH-induced erythrocyte hemolysis by scavenging free radicals at the cellular level. It caused the restoration of intracellular antioxidant enzyme (GSH, CAT and SOD) activities at an acceptable pace and the silencing of intracellular MDA formation. Furthermore, MTT toxicity assay showed that the novel antioxidant peptides had good biocompatibility and no cytotoxicity. Molecular docking and molecular dynamics simulations were used to investigate the molecular mechanism. The results showed that Ser363, Arg380, Asn414 and Arg415 residues played key roles in the antioxidant activity of MHW, YHW and FHW. And they mainly binded to the important residues of Keap1 protein stably by electrostatic force, van der Waals force, H-bonds and hydrophobic interactions, which can better fit the Kelch pocket structural domain of Keap1. Therefore, MHW, YHW and FHW had strong potential to interfere with Keap1-Nrf2 interaction and promoted the enzymatic expression of Nrf2-ARE downstream target pathway.

Screening and Functional Analyses of Novel Cecropins from Insect Transcriptome.

Antibiotic resistance is a significant and growing threat to global public health. However, antimicrobial peptides (AMPs) have shown promise as they exhibit a broad spectrum of antibacterial activities with low potential for resistance development. Insects, which inhabit a wide range of environments and are incredibly diverse, remain largely unexplored as a source of novel AMPs. To address this, we conducted a screening of the representative transcriptomes from the 1000 Insect Transcriptome Evolution (1KITE) dataset, focusing on the homologous reference genes of Cecropins, the first identified AMPs in insects known for its high efficiency. Our analysis identified 108 Cecropin genes from 105 insect transcriptomes, covering all major hexapod lineages. We validated the gene sequences and synthesized mature peptides for three identified Cecropin genes. Through minimal inhibition concentration and agar diffusion assays, we confirmed that these peptides exhibited antimicrobial activities against Gram-negative bacteria. Similar to the known Cecropin, the three Cecropins adopted an alpha-helical conformation in membrane-like environments, efficiently disrupting bacterial membranes through permeabilization. Importantly, none of the three Cecropins demonstrated cytotoxicity in erythrocyte hemolysis tests, suggesting their safety in real-world applications. Overall, this newly developed methodology provides a high-throughput bioinformatic pipeline for the discovery of AMP, taking advantage of the expanding genomic resources available for diverse organisms.

Protective Effects of Mulberry (Morus atropurpurea Roxb.) Leaf Protein Hydrolysates and Their In Vitro Gastrointestinal Digests on AAPH-Induced Oxidative Stress in Human Erythrocytes.

Mulberry leaf protein hydrolysates (HMP), and their in vitro gastrointestinal digests (GHMP), have shown favorable chemical antioxidant activities. The aim of this study is to investigate the potential protective effects of HMP and GHMP against 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced oxidative stress in human erythrocytes. The inhibition rate of hemolysis, the reactive oxygen species (ROS) level, the concentration of malondialdehyde (MDA), the reduced glutathione (GSH) and oxidized glutathione (GSSH), and the enzymatic activities of total superoxide dismutase (SOD), catalase (CAT), and cellular glutathione peroxidase (GSH-Px) were evaluated as the biomarkers of oxidative status in human erythrocytes. The results showed that HMP and GHMP effectively inhibit the occurrence of erythrocyte hemolysis in the range of 0.025-1.0 mg/mL, and the inhibition rates of HMP and GHMP reached 92% and 90% at concentrations of 0.4 mg/mL and 1.0 mg/mL, respectively. HMP and GHMP reduced the AAPH-induced oxidative hemolysis damage via suppressing the generation of ROS by inhibiting the formation of MDA, maintaining the balance of GSH/GSSG, and preserving the activities of the antioxidant enzymes, including SOD, GSH-Px, and CAT. Our findings revealed that both HMP and GHMP could be used as natural antioxidants, and have the potential for further application in the development of functional foods.

Anti-Hyperuricemic, Anti-Arthritic, Hemolytic Activity and Therapeutic Safety of Glycoconjugated Triazole-Phthalimides.

Hyperuricemia, the metabolic alteration that leads to gout or gouty arthritis, is increasing worldwide. Glycoconjugated triazole-phthalimides show potent anti-inflammatory activity. The aim of this study was to evaluate the anti-hyperuricemia effect of glycoconjugated triazole-phthalimides. To develop hyperuricemia, groups of mice received orally potassium oxonate (250 mg/kg) for 7 days, and F2, F3 and F4 glycoconjugated triazole-phthalimides (20 mg/kg), allopurinol (300 mg/kg), and 1% carboxymethylcellulose; indomethacin (2 and 4 mg/kg) was the positive control for anti-arthritic effect. Genotoxic and mutagenic effects were evaluated by the comet and micronucleus assays, respectively. The hemolytic action of the compounds was evaluated. Phthalimides F2, F3 and F4 significantly reduced the levels of serum uric acid, creatinine and urea in hyperuricemic animals. In addition, the compounds were efficient in reducing protein denaturation in a dose-dependent manner. In an interesting way, the histopathological analysis of kidneys from groups treated with F2, F3 and F4 showed a glomerular architecture, with the Bowman's capsule and renal tubules having a normal appearance and without inflammatory changes. Also, F2 and F4 showed a small increase in micronuclei, indicating a low mutagenic effect, whilst by comet assay only, we could infer that F4 affected the frequency and damage index, thus indicating a very small genotoxic action. Similarly, the phthalimides showed a low degree of erythrocyte hemolysis (<3%). Our data demonstrate that the new glycoconjugate triazole-phthalimides have potential to treat hyperuricemia and its secondary complications, such as gouty arthritis, with a low to non-significant rate of erythrocytes hemolysis, genotoxicity and mutagenicity making these molecules strong candidates as pharmaceutical agents for treatment requiring uric-acid-lowering therapy.

Beta-adrenoreactivity of erythrocyte membranes and expression of beta1-adrenergic receptors of cardiomyocytes in patients with heart failure with different left ventricular ejection fraction

Introduction. More and more studies demonstrate the importance of determining the beta-adrenergic reactivity of erythrocyte membranes (beta-ARM) for predicting the severity of cardiovascular diseases, including chronic heart failure. But the presence of general patterns between the change in beta-ARM and the expression of beta1adrenergic receptors on the membranes of cardiomyocytes, depending on the left ventricular ejection fraction (LVEF) remains questionable.Aim: To evaluate beta-ARM and expression of beta1-adrenergic receptors of cardiomyocytes in groups of patients with heart failure with preserved (≥ 50%) and reduced (&lt; 50%) LV EF.Materials and methods. The study included 100 patients with chronic heart failure of ischemic origin. Groups of 79 patients with LVEF ≥ 50% and 21 patients with LVEF &lt; 50% were formed. The levels of epinephrine and norepinephrine in blood plasma and the expression of beta1-adrenergic receptors in myocardial samples were assessed using enzyme immunoassay. Beta-ARM was determined in whole blood by assessing the inhibition of hemolysis of erythrocytes placed in a hypoosmotic medium in the presence of a selective beta-blocker.Results. In the group with LV EF &lt; 50%, beta-ARM was increased (p = 0.037), amounting to 31.9% (23.2; 40.1), compared with the group with LVEF ≥ 50%, where beta-ARM was 21.4% (15.0; 31.7). At the same time, in patients with LV EF &lt; 50%, the expression of beta1-AR on cardiomyocyte membranes was significantly lower than in the other group (p = 0.037): 2.7 (2.1; 3.0) ng/mg and 3.1 (2.4; 3.9) ng/mg total protein, respectively. However, plasma levels of epinephrine and noradrenaline were comparable in the study groups.Conclusion. Patients with heart failure with LVEF &lt; 50% were characterized by increased beta-ARM and reduced expression of beta1-AR on cardiomyocyte membranes compared with the group with LVEF ≥ 50% with comparable levels of circulating catecholamines.

Mitapivat: A Quinolone Sulfonamide to Manage Hemolytic Anemia in Adults With Pyruvate Kinase Deficiency.

Pyruvate kinase (PK) deficiency is a rare enzyme-linked glycolytic defect resulting in mild-to-severe chronic persistent erythrocyte hemolysis. The disease is an autosomal recessive trait caused by mutations in the PK liver and red blood cell gene characterized by insufficient erythrocyte PK activity. PK deficiency is most diagnosed in persons of northern European descent and managed with packed red blood cell transfusions, chelation, and splenectomy with cholecystectomy. Mitapivat is the first approved therapy indicated for hemolytic anemia in adults with PK deficiency with the potential for delaying splenectomy in mild-moderate disease. Mitapivat is a PK activator that acts by allosterically binding to the PK tetramer and increases PK activity. The red blood cell form of PK is mutated in PK deficiency, which leads to reduced adenosine triphosphate, shortened red blood cell lifespan, and chronic hemolysis. The half-life of elimination is 3-5 hours, with 73% bioavailability, 98% plasma protein binding, and a median duration of response of 7 months. Mitapivat has been investigated through various clinical trials for different therapeutic indications. Pivotal trials that serve the primary focus throughout this article are ACTIVATE, ACTIVATE-T, and RISE. ACTIVATE is a phase 3, randomized, double-blind, placebo-controlled study that evaluated the efficacy and safety of mitapivat in adult patients who were not receiving regular blood transfusions. Contrarily, ACTIVATE-T explored the safety and efficacy of mitapivat in adults with PK deficiency who received regular blood transfusions. Both trials demonstrated favorable use of mitapivat in PK deficiency. Focusing on another indication, the ongoing RISE trial investigates the optimal dosage of mitapivat in sickle cell disease. Mitapivat is an appropriate treatment for adults with PK deficiency requiring transfusions and may be considered for patients with symptomatic anemia who do not require transfusions and/or PK deficiency with compensated hemolysis without overt anemia.

Chemico-pharmacological and computational studies of Ophiorrhiza fasciculata D. Don and Psychotria silhetensis Hook. f. focusing cytotoxic, thrombolytic, anti-inflammatory, antioxidant, and antibacterial properties

The current study sought to examine the pharmacological potentials of crude methanolic extracts of Ophiorrhiza fasciculata and Psychotria silhetensis, as well as their various solvent fractionates, with a focus on cytotoxic, thrombolytic, membrane stabilizing, antioxidant, and antibacterial activities via in vitro and in silico approaches. The extensive chromatographic and spectroscopic analyses confirmed and characterized two compounds as (±)-licarin B (1) and stigmasterol (2) from O. fasciculata and P. silhetensis, respectively. Petroleum ether soluble fraction of O. fasciculata and the aqueous soluble fraction of P. silhetensis showed the lowest 50% lethal concentrations (1.41 and 1.94 μg/mL, respectively) in brine shrimp bioassay. Likewise, petroleum ether soluble fraction of O. fasciculata and aqueous soluble fraction of P. silhetensis showed the highest thrombolytic activity with 46.66% and 50.10% lyses of the clot, respectively. The methanol and dichloromethane soluble fractions of O. fasciculata reduced erythrocyte hemolysis by 64.03% and 37.08%, respectively, under hypotonic and heat-induced conditions, compared to 81.97% and 42.12% for standard acetylsalicylic acid. In antioxidant activity test, aqueous soluble fraction O. fasciculata (IC50 = 7.22 μg/mL) revealed promising antioxidant potentialities in comparison to standard butylated hydroxytoluene (IC50 = 21.20 μg/mL). In antibacterial screening, chloroform, and dichloromethane soluble fractions of P. silhetensis showed a mild antibacterial activity compared with the standard drug ciprofloxacin. Additionally, the molecular docking study corroborated the current in vitro findings, and the isolated two constituents had higher binding affinities toward epidermal growth factor receptor, tissue plasminogen activator, vFLIP-IKK gamma stapled peptide dimer, glutathione reductase, and dihydrofolate reductase enzyme than their corresponding standard drugs. In addition, the both isolated compounds exerted favorable pharmacokinetics (absorption, distribution, metabolism, excretion) and toxicological profiles with drug-like qualities in computational-based ADMET and drug likeliness analyses. The current research suggests that both plants have potential as a natural treatment for treating thrombosis, inflammation, and oxidative stress. However, more thorough research is required to thoroughly screen for phytochemicals and pinpoint the precise mechanisms of action of the bioactive metabolites derived from these plants against a broad range of molecular targets.

Long-Term L-Glutamine Treatment Reduces Hemolysis without Ameliorating Hepatic Vaso-Occlusion and Liver Fibrosis in a Mouse Model of Sickle Cell Disease.

Sickle cell disease (SCD) is an autosomal recessive monogenic disorder caused by a homozygous mutation in the β-globin gene, which leads to erythrocyte sickling, hemolysis, vaso-occlusion, and sterile inflammation. The administration of oral L-glutamine has been shown to reduce the frequency of pain in SCD patients; however, the long-term effect of L-glutamine in SCD remains to be determined. To understand the long-term effect of L-glutamine administration in the liver we used quantitative liver intravital microscopy and biochemical analysis in humanized SCD mice. We here show that chronic L-glutamine administration reduces hepatic hemoglobin-heme-iron levels but fails to ameliorate ischemic liver injury. Remarkably, we found that this failure in the resolution of hepatobiliary injury and persistent liver fibrosis is associated with the reduced expression of hepatic Kupffer cells post-L-glutamine treatment. These findings establish the importance of investigating the long-term effects of L-glutamine therapy on liver pathophysiology in SCD patients.

Study on the antioxidant activity of peptides from soybean meal by fermentation based on the chemical method and AAPH-induced oxidative stress.

Preparation and antioxidant activities of soybean peptides using solid fermentation to decrease the content of trypsin inhibitor (TI) and antigen protein were investigated in this study. The results showed the optimal fermentation conditions were as follows: fermentation time 48 h, the ratio of material to solvent 1:2, inoculum size 12%, and the ratio of Lactic acid bacteria and Aspergillus oryzae 2:1. The hydrolysate was were divided into four components of <1, 1-3, 3-5, and >5 kDa by ultrafiltration based on molecular weight, and the <1 kDa peptides expressed the highest antioxidant activities. Meanwhile, the cell antioxidant activity of the <1 kDa soybean peptides was investigated using AAPH-induced erythrocyte hemolysis, which effectively inhibited erythrocyte hemolysis with the inhibit rate of 85.8% through inhibition of the ROS intracellular generation. In addition, soybean peptides could significantly restore the intracellular antioxidant enzymes (SOD, GSH-Px, and CAT) activities, as well as inhibited intracellular MDA generation and depletion of GSH. The intracellular antioxidant detoxifying mechanism of soybean peptides was associated with both non-enzymatic and enzymatic defense systems. According to this study, fermentation could effectively improve the antioxidant activities of soybean peptides.

Kinetics and mechanisms of oxidative hemolysis of erythrocytes in the presence of azoand peroxide initiator

The kinetics of oxidative hemolysis of a 0.2% suspension of mouse erythrocytes in the presence of radicalforming initiators, 2,2'-azobis(2-amidinopropane) dihydrochloride and tert-butyl hydroperoxide, was studied over a wide range of concentrations. Hemolysis of erythrocytes proceeded with the activation of lipid peroxidation in erythrocyte membranes. In the case of tert-butyl hydroperoxide, oxidative processes also developed in the soluble part of the cell, leading to the formation of insoluble hemoglobin aggregates. An induction period of hemolysis (the time period required to reach 10% hemolysis) in the presence of 2,2'-azobis(2-amid-inopropane) dihydrochloride decreases inversely proportional to the square root of the initiator concentration, that is in accord with the classical theory of radical chain oxidation of hydrocarbons. In the case of tertbutyl hydroperoxide, this association was not found. The use of 2,2'-azobis(2-amidinopropane) dihydrochloride as a lipid peroxidation initiator is preferred in the erythrocyte model for testing natural and synthetic compounds for antioxidant activity.

Hemolysin from Aeromonas hydrophila enhances the host's serum enzyme activity and regulates transcriptional responses in the spleen of Cyprinus rubrofuscus

Aeromonas hydrophila is a conditional pathogen impacting public hygiene and safety. Hemolysin is a virulence factor of Aeromonas hydrophila that causes erythrocyte hemolysis, yet its transcriptional response to Cyprinus rubrofuscus remains unknown. Our investigation confirmed the hemolysis of hemolysin from A. hydrophila. Serum enzyme activity was evaluated weekly after C. rubrofuscus were immunized with hemolysin Ahh1. The results showed that the hemolysin enhances the serum superoxide dismutase (SOD), lysozyme (LZM), and catalase (CAT) activity, which reached a maximum on day 14. To elucidate the molecular interaction between hemolysin from A. hydrophila and the host, we performed transcriptome sequencing on the spleen of C. rubrofuscus 14 days post hemolysin infection. The total number of clean reads was 41.37 Gb, resulting in 79,832 unigenes with an N50 length of 1863 bp. There were 1982 significantly differentially expressed genes (DEGs), including 1083 upregulated genes and 899 downregulated genes. Transcript levels of the genes, such as LA6BL, CD2, and NLRC5, were significantly downregulated, while those of IL11, IL1R2, and IL8 were dramatically upregulated. The DEGs were mainly enriched in the immune disease, viral protein interaction with cytokine and cytokine receptor, and toll-like receptor pathways, suggesting that hemolysin stimulation can activate the transcriptional responses. RT-qPCR experiments results of seven genes, IL-8, STAT2, CTSK, PRF1, CXCL9, TLR5, and SACS, showed that their expression was highly concordant with RNA-seq data. We clarified for the first time the key genes and signaling pathways response to hemolysin from A. hydrophila, which offers strategies for treating and preventing diseases.

Synthesis and some properties of new bromo derivatives of isobornylphenols

We determined optimal conditions to brominate various isobornyl phenols with molecular bromine and N -bromosuccinimide as well as studied oxidative bromination of them with KBr-Oxone®. For obtained brominated derivatives, as isobornylphenols, an assessment of erythrotoxicity, membrane-protective and antioxidant activity was performed using in vitro models. Halogen derivatives of isobornylphenols containing a bromine atom in the para -position and an alkyl fragment in the ortho -position relative to the phenolic hydroxyl group showed high antioxidant activity in the model of oxidative hemolysis of erythrocytes.

Evaluation of In-vitro Antioxidant Potential of Bryophillum pinnatum Leaf Juice in Phenyl Hydrazine Treated Goat Erythrocytes

Phytochemicals are known to contain a high concentration of therapeutically active principles, which, when consumed in sufficient quantities, can have drug-like effects. Bryophillum pinnatum (Lam.) is a succulent-ethno medicinal xerophyte with a long history of use in traditional Indian medicine. Because reactive oxygen species are the most common etiological factor in many degenerative diseases, controlling them with antioxidants may both treat and prevent disease progression. The current study will look at the ability of fresh leaf juice of Bryophillum pinnatum (Lam.) (FLJBP) to prevent oxidative injury and subsequent hemolysis in goat erythrocytes as a model test system. The catalytic activity of membrane bound enzymes like sodium potassium-ATPase, Ach-Ease, and metabolic enzymes like hexokinase, as well as the NO concentration in goat RBCs, were measured. Phenyl hydrazine was used first, followed by FLJBP at concentrations of 20, 40, 60, 80, and 100g/ml. A dose-dependent (PHZ) increase in Pi liberation by hexokinase and Na+/K+ ATPase, as well as increased formation of thiocholine iodide and enhanced NO scavenging potential, has indicated that flip has good anti-oxidant and anti-haemolytic activity. To record the functional food status of Bryophillum pinnatum (Lam.) species, a more rigorous and stringent battery of pharmacological, phytochemical, and bio analytical studies, followed by observational studies in humans, will be conducted.

Gender-related Characteristics of Oxidative Stress Severity and Cognitive Impairment in Patients With Schizophrenia.

The gender specificity of the clinical and psychopathological features of schizophrenia is an important factor in this disease. Gender features of neurocognitive deficit in schizophrenia and associated functional and structural disorders of the brain activity are of particular interest to researchers. There are several potential pathogenetic factors of this disease associated with gender, one of which is considered to be oxidative stress. Stress-induced cell death in the prefrontal and anterior frontal regions and reduced brain volume in these regions lead to cognitive and executive decline in patients with schizophrenia. Despite the great interest in the gender factor in schizophrenia pathogenesis, there are currently very few studies on gender differences in the severity of redox imbalance in patients with schizophrenia and their association with neurocognitive deficit.The aim of the study was to reveal the gender specificity of oxidative stress severity in patients with schizophrenia and assess its association with the severity of neurocognitive deficit.The study included 125 patients with schizophrenia and 75 sex- and age-matched healthy volunteers. Sociodemographic and clinical data were studied. Cognitive functions were evaluated using BACS. Blood samples were taken for biochemical studies of the levels of reduced glutathione (GSH), superoxide dismutase (SOD), catalase (CAT) in erythrocyte hemolysate, malonic dialdehyde (MDA), aldehyde-2,4-dinitrophenylhydrazones (ADNPH), ketone-2,4-dinitrophenylhydrazones (KDNPH) in plasma. Levels of oxidative stress markers were assessed by spectrophotometric method.In a sample of patients with schizophrenia, a statistically significantly higher activity of CAT was revealed among women compared to men (T=2.25; p=0.025), however, it was lower than in healthy volunteers. But, at the same time, a higher concentration of protein peroxidation products was found in the peripheral blood of women than in men (ADNPH MCO: T=2.52; p=0.013; KDNPH MCO: Z=-2.26; p=0.017). In the group of healthy volunteers, in contrast to patients with schizophrenia, gender differences in markers of oxidative stress were not found. In women with schizophrenia, single correlations were found between the level of the lipid peroxidation product MDA and the test scores for verbal memory (R=-0.36; p=0.006) and working memory (R=-0.36; p=0.006), antioxidant enzyme activity SOD and motor skills (R=-0.26; p=0.047). In men, on the contrary, multiple correlations of both antioxidants and, mainly, products of lipid and protein peroxidation with cognitive functions assessed using BACS were found.Despite the fact that oxidative stress is more pronounced in women than in men with schizophrenia, associations of redox imbalance with neurocognitive deficit in women is much less pronounced than in men.

Construction of a visualized liver-targeting siRNA delivery system

A visualized liver-targeting small-interfering RNA (siRNA) delivery carrier, glycyrrhetinic acid-modified lipid-[poly(aspartate-graft-polyethyleneimine)]-modified superparamagnetic nanoparticles (GLPPS), was prepared for focus on improving the siRNA delivery efficiency and diagnostic capability for hepatocellular carcinoma. Agarose gel electrophoresis, cytotoxicity, and in vitro gene silencing experiments were conducted to determine the optimal carrier formulation. The results showed that the best formulation of the carrier was a weight ratio of LPPS to siRNA of 30:1 and a molar ratio of LPPS to DSPE-PEG-GA of 65:3. Prussian blue staining and in vitro magnetic resonance imaging (MRI) experiments further confirmed that GLPPS had a significant liver-targeting effect. The carrier's biocompatibility was tested both in vitro and in vivo, and the results showed no hemolysis or morphological changes of erythrocytes when the carrier was incubated with rabbit blood cells. Following injection of GLPPS into the tail vein of mice, the hematoxylin and eosin staining of the liver, spleen, and kidney showed normal tissue morphology with no inflammation or injury. Taken together, these results indicated that GLPPS is a safe drug carrier. In vivo MRI showed that GLPPS significantly reduced tumor signal intensity and improved MRI contrast. In conclusion, GLPPS may be a promising liver-targeting siRNA delivery and MRI monitoring carrier.

Kinetics and Mechanisms of Oxidative Hemolysis of Erythrocytes under the Action of Azo- and Peroxide Initiator

Kinetics and Mechanisms of Oxidative Hemolysis of Erythrocytes under the Action of Azo- and Peroxide Initiator

Comparative transcriptomics analysis of multidrug-resistant Acinetobacter baumannii in response to treatment with the terpenic compounds thymol and carvacrol

Acinetobacter baumannii is a gram-negative opportunistic bacterium that has become a major public health concern and a substantial medical challenge due to its ability to acquire multidrug resistance (MDR), extended-drug resistance, or pan-drug resistance. In this study, we evaluated the antibacterial activity of thymol and carvacrol alone or in combination against clinical isolates of MDR A. baumannii. Additionally, we used RNA-sequency to perform a comparative transcriptomic analysis of the effects of carvacrol and thymol on the Acb35 strain under different treatment conditions. Our results demonstrated that thymol and carvacrol alone, effectively inhibited the bacterial growth of MDR A. baumannii isolates, with a minimum inhibitory concentration (MIC) lower than 500 μg/mL. Furthermore, the combination of thymol and carvacrol exhibited either synergistic (FICI ≤ 0.5) or additive effects (0.5 < FICI ≤ 4), enhancing their antibacterial activity. Importantly, these compounds were found to be non-cytotoxic to Vero cells and did not cause hemolysis in erythrocytes at concentrations that effectively inhibited bacterial growth. Transcriptomic analysis revealed the down-regulation of mRNA associated with ribosomal subunit assemblies under all experimental conditions tested. However, the up-regulation of specific genes encoding stress response proteins and transcriptional regulators varied depending on the experimental condition, particularly in response to the treatment with carvacrol and thymol in combination. Based on our findings, thymol and carvacrol demonstrate promising potential as chemotherapeutic agents for controlling MDR A. baumannii infections. These compounds exhibit strong antibacterial activity, particularly in combination and lower cytotoxicity towards mammalian cells. The observed effects on gene expression provide insights into the underlying mechanisms of action, highlighting the regulation of stress response pathways.