Generation Of Species Research Articles

Transient neonatal exposure to hyperoxia, an experimental model of preterm birth, leads to skeletal muscle atrophy and fiber type switching.

Individuals born preterm show reduced exercise capacity and increased risk for pulmonary and cardiovascular diseases, but the impact of preterm birth on skeletal muscle, an inherently critical part of cardiorespiratory fitness, remains unknown. We evaluated the impacts of preterm birth-related conditions on the development, growth, and function of skeletal muscle using a recognized preclinical rodent model in which newborn rats are exposed to 80% oxygen from days 3 to 10 of life. We analyzed different hindlimb muscles of male and female rats at 10 days (neonatal), 4 weeks (juvenile), and 16 weeks (young adults). Neonatal high oxygen exposure increased the generation of reactive oxygen species (ROS) and the signs of inflammation in skeletal muscles, which was associated with muscle fiber atrophy, fiber type shifting (reduced proportion of type I slow fibers and increased proportion of type IIb fast-fatigable fibers), and impairment in muscle function. These effects were maintained until adulthood. Fast-twitch muscles were more vulnerable to the effects of hyperoxia than slow-twitch muscles. Male rats, which expressed lower antioxidant defenses, were more susceptible than females to oxygen-induced myopathy. Overall, preterm birth-related conditions have long-lasting effects on the composition, morphology, and function of skeletal muscles; and these effects are sex-specific. Oxygen-induced changes in skeletal muscles could contribute to the reduced exercise capacity and to increased risk of diseases of preterm born individuals.

Toxicity of stainless and mild steel particles generated from gas\u2013metal arc welding in primary human small airway epithelial cells

Welding fumes induce lung toxicity and are carcinogenic to humans but the molecular mechanisms have yet to be clarified. The aim of this study was to evaluate the toxicity of stainless and mild steel particles generated via gas–metal arc welding using primary human small airway epithelial cells (hSAEC) and ToxTracker reporter murine stem cells, which track activation of six cancer-related pathways. Metal content (Fe, Mn, Ni, Cr) of the particles was relatively homogenous across particle size. The particles were not cytotoxic in reporter stem cells but stainless steel particles activated the Nrf2-dependent oxidative stress pathway. In hSAEC, both particle types induced time- and dose-dependent cytotoxicity, and stainless steel particles also increased generation of reactive oxygen species. The cellular metal content was higher for hSAEC compared to the reporter stem cells exposed to the same nominal dose. This was, in part, related to differences in particle agglomeration/sedimentation in the different cell media. Overall, our study showed differences in cytotoxicity and activation of cancer-related pathways between stainless and mild steel welding particles. Moreover, our data emphasizes the need for careful assessment of the cellular dose when comparing studies using different in vitro models.

Potential therapeutic approaches in paraquat poisoning: a narrative review

Paraquat dichloride (PQ) poisoning is a relatively rare yet critical medical condition that has a high case fatality rate. Lung tissue is highly susceptible to PQ-induced injury, and respiratory failure is the leading cause of death in these patients. Unfortunately, there is a lack of an effective therapeutic approach to ameliorate outcomes. It is well-known that PQ interferes with a variety of cell signaling pathways and induces the generation of reactive oxygen species (ROS), which ultimately results in cell injury. The traditional treatment decisions have not been able to significantly change the clinical course of PQ poisoning. Moreover, novel therapeutic strategies for PQ poisoning have centered on the inhibition of PQ-induced signaling pathways. In the current review, we sought to provide a bird’s-eye view of the available therapeutic approaches in patients with PQ poisoning.

YBX1 knockdown induces renal cell carcinoma cell apoptosis via Kindlin-2

ABSTRACT Among urological tumors, renal cell carcinoma (RCC) is the third-highest mortality rate tumor, and 20%–30% of RCC patients present with metastases at the time of diagnosis. While the treatment of RCC has been improved over the last few years, its mortality stays high. Y-box binding protein 1 (YBX1) is a well-known oncoprotein that has tumor-promoting functions. YBX1 is widely considered to be an attractive therapeutic target in cancer. To develop novel therapeutics to target YBX1, it is of great importance to understand how YBX1 is finely regulated in cancer. Our previous studies showed that YBX1 in RCC cells significantly promoted cell adhesion, migration, and invasion. However, the role of YBX1 in RCC cells apoptosis has not been reported. In this study, we investigated the effect of YBX1 on cell apoptosis and elucidated the mechanisms involved. Results showed that YBX1 regulated RCC cells apoptosis and reactive oxygen species (ROS) generation via Kindlin-2. These findings indicated that YBX1 inhibited RCC cells apoptosis and may serve as a candidate RCC prognostic marker and a potential therapeutic target. Abbreviations: RCC: Renal cell carcinoma; YBX1: Y-box binding protein 1; ROS: Reactive oxygen species; ccRCC: Clear cell renal cell carcinoma; mccRCC: Metastatic clear cell renal cell carcinoma; G3BP1: Ras-GTPase activating protein SH3 domain-binding proteins 1; SPP1: Secreted phosphoprotein 1; NF-κB: Nuclear factor kappa beta; ECM: Extracellular matrix; EMT: Epithelial-mesenchymal transition; PYCR1: Pyrroline-5-carboxylate reductase 1; MEM: Eagle’s Minimum Essential Medium; DMEM: Dulbecco’s modified Eagle medium; FBS: Fetal bovine serum; PCR: Polymerase chain reaction; shRNA: Short hairpin RNA; siRNA: Small interfering RNA; BSA: Bovine serum albumin; DCFH-DA: 2,7-Dichlorodihydrofluorescein diacetate; FITC: Fluorescein isothiocyanate; PI: Propidium iodide;

Hydrogenated Amorphous TiO2-x and Its High Visible Light Photoactivity.

Hydrogenated crystalline TiO2 with oxygen vacancy (OV) defect has been broadly investigated in recent years. Different from crystalline TiO2, hydrogenated amorphous TiO2−x for advanced photocatalytic applications is scarcely reported. In this work, we prepared hydrogenated amorphous TiO2−x (HA-TiO2−x) using a unique liquid plasma hydrogenation strategy, and demonstrated its highly visible-light photoactivity. Density functional theory combined with comprehensive analyses was to gain fundamental understanding of the correlation among the OV concentration, electronic band structure, photon capturing, reactive oxygen species (ROS) generation, and photocatalytic activity. One important finding was that the narrower the bandgap HA-TiO2−x possessed, the higher photocatalytic efficiency it exhibited. Given the narrow bandgap and extraordinary visible-light absorption, HA-TiO2−x showed excellent visible-light photodegradation in rhodamine B (98.7%), methylene blue (99.85%), and theophylline (99.87) within two hours, as well as long-term stability. The total organic carbon (TOC) removal rates of rhodamine B, methylene blue, and theophylline were measured to 55%, 61.8%, and 50.7%, respectively, which indicated that HA-TiO2−x exhibited high wastewater purification performance. This study provided a direct and effective hydrogenation method to produce reduced amorphous TiO2−x which has great potential in practical environmental remediation.

Antimicrobial activity and mode of action of 1,8-cineol against carbapenemase-producing Klebsiella pneumoniae

Antimicrobial resistance remains one of the most challenging issues that threatens the health of people around the world. Plant-derived natural compounds have received considerable attention for their potential role to mitigate antibiotic resistance. This study was carried out to assess the antimicrobial activity and mode of action of a monoterpene, 1,8-cineol (CN) against carbapenemase-producing Klebsiella pneumoniae (KPC-KP). Results showed that resazurin microplate assay and time-kill analysis revealed bactericidal effects of CN at 28.83 mg/mL. Zeta potential showed that CN increased the surface charge of bacteria and an increase of outer membrane permeability was also detected. CN was able to cause leakage of proteins and nucleic acids in KPC-KP cells upon exposure to CN and ethidium bromide influx/efflux experiment showed the uptake of ethidium bromide into the cell; this was attributed to membrane damage. CN was also found to induce oxidative stress in CN-treated KPC-KP cells through generation of reactive oxygen species which initiated lipid peroxidation and thus damaging the bacterial cell membrane. Scanning and transmission electron microscopies further confirmed the disruption of bacterial cell membrane and loss of intracellular materials. In this study, we demonstrated that CN induced oxidative stress and membrane damage resulting in KPC-KP cell death.

AGSE: A Novel Grape Seed Extract Enriched for PP2A Activating Flavonoids That Combats Oxidative Stress and Promotes Skin Health.

Environmental stimuli attack the skin daily resulting in the generation of reactive oxygen species (ROS) and inflammation. One pathway that regulates oxidative stress in skin involves Protein Phosphatase 2A (PP2A), a phosphatase which has been previously linked to Alzheimer’s Disease and aging. Oxidative stress decreases PP2A methylation in normal human dermal fibroblasts (NHDFs). Thus, we hypothesize agents that increase PP2A methylation and activity will promote skin health and combat aging. To discover novel inhibitors of PP2A demethylation activity, we screened a library of 32 natural botanical extracts. We discovered Grape Seed Extract (GSE), which has previously been reported to have several benefits for skin, to be the most potent PP2A demethylating extract. Via several fractionation and extraction steps we developed a novel grape seed extract called Activated Grape Seed Extract (AGSE), which is enriched for PP2A activating flavonoids that increase potency in preventing PP2A demethylation when compared to commercial GSE. We then determined that 1% AGSE and 1% commercial GSE exhibit distinct gene expression profiles when topically applied to a 3D human skin model. To begin to characterize AGSE’s activity, we investigated its antioxidant potential and demonstrate it reduces ROS levels in NHDFs and cell-free assays equal to or better than Vitamin C and E. Moreover, AGSE shows anti-inflammatory properties, dose-dependently inhibiting UVA, UVB and chemical-induced inflammation. These results demonstrate AGSE is a novel, multi-functional extract that modulates methylation levels of PP2A and supports the hypothesis of PP2A as a master regulator for oxidative stress signaling and aging in skin.

Inhibition of ceramide synthesis improves the outcome in ischemia/reperfusion injury using human-induced pluripotent stem cell derived cardiomyocyte

Abstract Introduction and aim Ceramides are proven to be biologically active in apoptosis, inflammation, mitochondrial dysfunction, and as a second messenger in various signaling pathways1. However, the data linking the role of ceramides in ischemia/reperfusion injury (I/R injury) are lacking. We aimed to establish an I/R injury model using human-induced pluripotent stem cell (hiPSC)-derived cardiomyocyte (CM) and to evaluate ceramide levels, ceramide synthesis pathway, and outcome of CM with inhibition of ceramide synthesis during I/R injury. Methods HiPSC technology has been used to generate functional human CMs to elucidate the underlying mechanisms of the pathophysiology of the human heart. Results In our model, we observed an increase of mRNA levels of genes regulating ceramide synthesis after 6 h of ischemia followed by 16 h reperfusion, such as SPTLC1 (1.1±0.08 vs 1.0, p=0.2), CerS2 (1.6±0.3 vs 1.0, p<0.001), CerS4 (1.3±0.1 vs 1.0, p=0.02), CerS5 (1.3±0.1 vs 1.0, p=0.03), and SMPD (1.6±0.1 vs 1.0, p=0.008) compared to control. Also, both long- and very long-chain ceramide species levels measured with mass spectrometry were increased significantly after 6 h ischemia followed by 16 h reperfusion compared to control (C14:0: 1,1±0.3 pmol/million cells vs 0,3±0,2 pmol/ million cells, p=0.02 and C24:1: 26,3±7,1 pmol/ million cells vs 9,6±3,4 pmol/ million cells, p=0.02). Inhibition of ceramide synthesis with Fumonisin B1 (FB1) significantly increased the viability after 6h of ischemia followed by 16 h of reperfusion compared to CMs incubated without inhibitors (32.2%±1.5% vs 26.9%±2.6%, p=0.04). Interestingly, we identified two mechanisms with which the viability improves after incubation with ceramide inhibitor. The first mechanism observed could be the restoration of both intracellular calcium baseline (control 29±1.2, I/R 55±5.7 and I/R with FB1 35.6±2.5, p<0,001) and peak (control 45.1±5.6, I/R 94.3±5.7 and I/R with FB1 56.5±7.5, p<0,001) levels to nearly the same levels as observed in control samples. A possible cause of increased calcium oscillations after 6 h of ischemia followed by 3 h of reperfusion in the first place could be an upregulation of the RyR2 levels detected by qPCR (2.5±0.4 vs control 1.0, p=0.008). The second mechanism of improving viability in I/R injury could be a decrease of generation of reactive oxygen species (ROS) detected by MitoSOX dye after incubation with FB1 inhibitor to nearly the same levels as observed in control (control 22±5.1, I/R 33.8±5.8 and I/R with FB1 30.7±5.9, p=0,06). Conclusion We conclude that ceramides have important implications in either mediating or causing injury and their inhibition improves the outcome of I/R injury by decreasing ROS generation and improving calcium oscillations. Funding Acknowledgement Type of funding sources: Public hospital(s). Main funding source(s): Jena University Hospital, Clinic for Internal Medicine 1Interdisciplinary Center for Clinical Research Jena

Persistent/progressive subclinical contrast-induced nephropathy – predictors and effects of parenteral hydration. A prospective study in patients undergoing coronary angioplasty

Abstract Background In most cases, contrast-induced nephropathy (CIN) appears to be a transient event. The pathophysiology of CIN, which includes renal vasoconstriction and hypoxia, generation of cytotoxic reactive oxygen species, and even direct tubular toxicity, suggests however that the tubular impairment typical for CIN may not be entirely transient and that persistent/progressive subclinical kidney dysfunction could actually occur. Purpose We aimed to evaluate the incidence, identify the predictors, and assess the impact of parenteral hydration on the occurrence of persistent/progressive subclinical CIN in patients undergoing coronary angioplasty. Methods A total of 71 patients scheduled for elective coronary angioplasty were randomized into two groups: Control (n=36) and HYD (n=35). Patients in the HYD group received prophylactic hydration with saline solution (1 ml/kg/h; 6 h prior to and 12 h after the procedure). Plasma levels of neutrophil gelatinase-associated lipocalin (NGAL), a specific marker of tubular impairment, were evaluated prior to, and 48 h and 1 month after the angioplasty procedure. Persistent/progressive subclinical CIN at 1 month post-angioplasty was defined as a significant increase in NGAL at 48 h after the procedure that persisted (i.e., did not change by more than 10%) or aggravated (i.e., increased by more than 10%) at 1 month follow-up. The ability of several factors (i.e., age, hypertension, diabetes, use of nephrotoxic drugs, baseline creatinine clearance, volume of contrast agent) to predict persistent/progressive CIN was evaluated. The ability of parenteral hydration to prevent persistent/progressive CIN was also assessed. Results According to the current criteria, CIN was present in 5 (7.0%) patients. Meanwhile, subclinical CIN was identified in 36 (50.7%) patients, of which 10 (27.7%) presented persistent and 15 (41.6%) progressive CIN at 1 month follow-up. The volume of contrast administered during angioplasty was the only factor that differed significantly between patients who developed persistent/progressive CIN and those who did not (178±17 mL vs. 130±10 mL, p=0.01). In ROC analysis, contrast volume ≥150 mL predicted persistent/progressive CIN with 77% sensitivity and 60% specificity (p<0.001). Prophylactic hydration was associated with a 75% lower risk of persistent/progressive CIN (RR 0.25; 95% CI 0.09–0.65; p=0.001). Conclusions Despite the relatively low incidence of CIN, subclinical CIN was identified in more than 50% of the study patients. More than 2/3 of these patients presented persistent or progressive subclinical CIN at 1 month follow-up. The clinical impact of this persistent/progressive subclinical CIN in patients undergoing multiple contrast administrations remains to be established. Our data indicate that reducing the contrast volume and/or using prophylactic parenteral hydration can significantly reduce persistent/progressive subclinical CIN. Funding Acknowledgement Type of funding sources: Public grant(s) – National budget only. Main funding source(s): This work was supported by a grant of the Romanian Ministry of Education and Research, CNCS-UEFISCDI

Short-Chain Fatty Acids Alleviate Hepatocyte Apoptosis Induced by Gut-Derived Protein-Bound Uremic Toxins.

Chronic kidney disease (CKD) is characterized with the influx of uremic toxins, which impairs the gut microbiome by decreasing beneficial bacteria that produce short-chain fatty acids (SCFAs) and increasing harmful bacteria that produce gut-derived protein-bound uremic toxins (PBUTs). This study aimed to assess the proapoptotic effects of three major gut-derived PBUTs in hepatocytes, and the effects of SCFAs on apoptosis phenotype in vitro. HepG2 (human liver carcinoma cells) and THLE-2 (immortalized human normal liver cells) cell line were incubated with 0, 2, 20, 200, 2000 μM p-cresol sulfate (PCS), indoxyl sulfate (IS), and hippuric acid (HA), respectively, for 24 h. Flow cytometry analysis indicated that three uremic toxins induced varying degrees of apoptosis in hepatocytes and HA represented the highest efficacy. These phenotypes were further confirmed by western blot of apoptosis protein expression [Caspase-3, Caspase-9, B-cell lymphoma 2 (Bcl-2), and Bcl-2-associated X protein (Bax)]. Human normal hepatocytes (THLE-2) are more sensitive to PBUTs-induced apoptosis compared with human hepatoma cells (HepG2). Mechanistically, extracellular HA could enter hepatocytes, increase reactive oxygen species (ROS) generation, and decrease mitochondrial membrane potential dose-dependently in THLE-2 cells. Notably, coculture with SCFAs (acetate, propionate, butyrate) for 24 h significantly improved HA-induced apoptosis in THLE-2 cells, and propionate (500 μM) represented the highest efficacy. Propionate reduction of apoptosis was associated with improving mitochondria dysfunction and oxidative stress in a manner involving reducing Caspase-3 expression, ROS production, and increasing the Bcl-2/Bax level. As such, our studies validated PBUTs accumulation might be an important cause of liver dysfunction in patients with CKD, and supplementation of SCFAs might be a viable way to protect the liver for patients with CKD.

Cytotoxicity and pro-inflammatory effect of GaSb thin films in L929 cells

Gallium antimonide (GaSb)-based devices operate efficiently in the infrared region. Investigating the toxicity of GaSb thin film is necessary for using embedded GaSb-based devices in living organisms. In this study, viability, oxidative stress, inflammatory responses, apoptosis induction and genotoxicity of GaSb were assayed using L929 cells following a 24 h exposure to GaSb. GaSb thin films were deposited on a quartz substrate using radio frequency (RF) magnetron sputtering. These films were soaked in cell culture medium to prepare test solutions. The viability of cells treated with the GaSb extract was lower than that of control cells. GaSb elicited little reactive oxygen species (ROS) generation. Tumor necrosis factor (TNF)-[Formula: see text] and interleukin (IL)-1[Formula: see text] levels were elevated in GaSb-treated cell culture supernatants. Apoptosis and genotoxicity were not evident following GaSb treatment. Overall, these results demonstrate the low toxicity of GaSb compared with previous studies examining arsenic-containing III–V materials, which is desirable for biological devices.

Biodegradable mesoporous manganese carbonate nanocomposites for LED light-driven cancer therapy via enhancing photodynamic therapy and attenuating survivin expression

Triple-negative breast cancer (TNBC) is one of the most daunting diseases, low toxicity and efficient approaches are in urgent demand. Herein, we developed degradable mesoporous manganese carbonate nanocubes (MnCO3 NCs), incorporated with survivin shRNA-expressing plasmid DNA (iSur-pDNA) and riboflavin (Rf), namely MRp NCs, for synergistic TNBC therapy. The MnCO3, itself, could generate O2 and CO2 under H2O2 and thus relieve the hypoxia and acidic tumor microenvironment (TME). Furthermore, the MnCO3 NCs exhibited high Rf loading capacity and iSur-pDNA delivery ability after polyethyleneimine modification. Specifically, MRp NCs decompose in TME, meanwhile they deprived the endogenous expression of survivin gene and significantly amplified the generation of reactive oxygen species after exposure to LED light, resulting in serious tumor destruction. The multifunctional MRp NCs with LED light-driven characters are able to provide a high efficiency, low toxicity and promising strategy for TNBC therapy.

Mast Cells Modulate the Immune Response and Redox Status of the Gastrointestinal Tract in Induced Venom Pathogenesis

The pathogenesis of Androctonus autralis hector (Aah) scorpion venom involved cellular and molecular mechanisms resulting in multi-organ dysfunction. However, little is reported about the effects of venom on the gastrointestinal axis. Mast cells (MCs) are known to play a crucial role in modulating immune response of the gut. This study aims to investigate the involvement of this cell type in venom-induced gastric and intestinal disorders in a time course (3 and 24h). The obtained results revealed that Aah scorpion venom induced inflammatory cell infiltration as shown by the increase of the myeloperoxidase and eosinophil peroxidase activities. Overexpression of the c-kit receptor (CD117) severely imbalanced the redox status with depletion of antioxidant systemic accompanied by gastrointestinal tissue damage. Moreover, an increased level of lactate dehydrogenase in the serum was correlated with tissue injuries. Pharmacological inhibition of MCs targeting tyrosine kinase (TK) reduces the generation of reactive oxygen species and normalizes catalase, and gluthation S-transferase activities to their physiological levels. In addition, histopathological alterations were restored after pretreatment with c-kit receptor inhibitor associated with a considerable reduction of MC density. Interestingly, obtained results indicate that MCs might be involved in gastric modulation and intestinal inflammation through c-kit signaling following sub-cutaneous Aah venom injection.

Functional Group Transformation in Naphthoquinones: Strategies for the Synthesis of Mono- and Bis(Amino-1,4-Naphthoquinones)

: 1,4-Naphthoquinones are a group of natural and synthetic substances that participate in various biochemical functions in most plants and animals, being involved in energy production, electron transport, blood coagulation, and generation of reactive oxygen species (ROS), among others. The performance of such biochemical functions is directly related to the functional groups linked to the 1,4-naphthoquinone scaffold, and the introduction of nitrogencontaining groups into this nucleus may amplify its biological activity against numerous targets. Therefore, several methods have been developed in the last decades for preparing these compounds, especially from 1,4-naphthoquinone derivatives. Considering these concepts, in this review, we aim to provide insights into the chemistry, chemical synthesis, semi-synthesis, pharmacological effects, and reactivity of low molecular weight mono- and bis(aminonaphthoquinones); we start by discussing the use of some of the most important 1,4-naphthoquinones as starting materials for the synthesis of amino-1,4-naphthoquinones (ANQs), examine the modification of ANQ derivatives, and conclude by providing an outlook on this field.

Immunity to stripe rust in wheat: A case study of a hypersensitive-response (HR)- independent resistance to Puccinia striiformis f. sp. tritici in Avocet-Yr15

Stripe (yellow) rust, caused by the biotrophic fungal pathogen Puccinia striiformis f. sp. tritici (Pst), is emerging as a serious threat to wheat (Triticum aestivum L.) production in many regions of North America. Genetic resistance to stripe rust is typically conditioned by the products of resistance genes that detect pathogen-associated molecular patterns and initiate a cascade of signalling events, culminating in the generation of reactive oxygen species (ROS) and a hypersensitive response (HR) that eventually suppress the pathogen. Complete resistance with no symptoms (immunity response) has also been observed, but less frequently, in certain Pst-wheat interactions. However, such HR-independent resistance responses to Pst have not been well studied. Here we report that the Pst-resistant near isogenic line Avocet-Yr15 (carrying the Yr15 resistance gene) exhibited an immunity response to an isolate of Pst collected in Elora, Ontario, while the Avocet-S line (lacking Yr15) was highly susceptible to the same isolate. In particular, histochemical assays of defence-associated ROS generation and HR were negative during the early stages of the interaction between Pst and the resistant Avocet-Yr15. Microscopic analysis indicated that the pathogen was not able to penetrate the stomata of Avocet-Yr15. Moreover, RNA-sequencing of the immunity response of Avocet-Yr15 to Pst highlighted the activation of a global anti-oxidative-stress response consisting of genes involved in the maintenance of cell viability, redox homoeostasis and photosynthesis. These results provide insights into the molecular mechanisms controlling the immunity response to stripe rust of wheat.

Potential Roles of Sestrin2 in Alzheimer's Disease: Antioxidation, Autophagy Promotion, and Beyond.

Alzheimer’s disease (AD) is the most common age-related neurodegenerative disease. It presents with progressive memory loss, worsens cognitive functions to the point of disability, and causes heavy socioeconomic burdens to patients, their families, and society as a whole. The underlying pathogenic mechanisms of AD are complex and may involve excitotoxicity, excessive generation of reactive oxygen species (ROS), aberrant cell cycle reentry, impaired mitochondrial function, and DNA damage. Up to now, there is no effective treatment available for AD, and it is therefore urgent to develop an effective therapeutic regimen for this devastating disease. Sestrin2, belonging to the sestrin family, can counteract oxidative stress, reduce activity of the mammalian/mechanistic target of rapamycin (mTOR), and improve cell survival. It may therefore play a crucial role in neurodegenerative diseases like AD. However, only limited studies of sestrin2 and AD have been conducted up to now. In this article, we discuss current experimental evidence to demonstrate the potential roles of sestrin2 in treating neurodegenerative diseases, focusing specifically on AD. Strategies for augmenting sestrin2 expression may strengthen neurons, adapting them to stressful conditions through counteracting oxidative stress, and may also adjust the autophagy process, these two effects together conferring neuronal resistance in cases of AD.

Вплив свинцю на репродуктивне здоров'я чоловіків

Lead does not succumb to biological decomposition, and its ability to accumulate in the body makes it a serious threat to the health of people and animals, while affecting the reproductive function. In most cases, poisoning with lead remains asymptomatic. In a number of studies the authors concluded that in men the level of lead in the blood more than >40 μg/dL leads to the disorder of reproductive functions, such as low libido, a small sperm volume, the amount of spermatozoa, an increase in the abnormal morphology of spermatozoa and decrease in their motility. Male factors are considered the main cause of infertility in 40% of infertile couples and contribute to the emergence of this state in combination with female factors in 20% of cases. The mechanisms of how lead causes male infertility are covered in depth. It is assumed that the basic effect on the reproductive function of men is likely to occur due to changes in the reproductive hormonal axis and hormonal control of spermatogenesis, and not due to direct toxic effects on the seminiferous tubules. The adverse effect of lead on the male reproductive function, especially at low doses (<10 μg/dl), has not been studied properly yet. The risk of lead poisoning is directly connected not only with an increase in concentration, but also with the duration of the impact of metal. There are a number of possible ways of how exposure of lead reduces male fertility. Lead, most likely, impairs the endocrine profile of regulation, mainly through the axis of the hypothalamus-pituitary testosterone, hereafter reduces the production of sperm in the seminiferous tubules of the testicles. At the same time, it acts as an endocrine destroyer, affecting hormones responsible for the production of sperm. In addition to changes in the reproductive hormone axis and hormonal control of spermatogenesis, the activity of enzymes, such as alkaline phosphatase and potassium-sodium ATP-ase, direct toxic effects on the seminiferous tubules, the exposure time of the metal and its dose affect male infertility. Another problem associated with the reproductive toxicity of lead is determined by the excessive generation of the reactive oxygen species. It is known that the oxidative stress caused by lead is involved in the abnormal functions of spermatozoa and male infertility. The possibility to decrease lead level in the body using a number of methods, such as chelatotherapy, nano-encapsulation, use of N-acetylcysteine is considered. Conclusion. Based on animal studies, it seems to be rational to prescribe the corresponding antioxidants to persons suffering from abnormal parameters of spermatozoa and infertility due to the effects of lead. Antioxidants showed a protective effect on spermatogenesis on animal models and reduced reactive oxygen species in sperm and DNA fragmentation in studies in humans. Although there is no final evidence confirming the use of antioxidant additives in men with low fertility to improve fertility rates, it is believed that due to the low cost and a small number of side effects, antioxidants need to be recommended to men with insufficient fertility

The Effect of Cold Atmospheric Plasma on the Expression of Apoptotic Genes in Breast Cancerous Cells

Background: Breast cancer is one of the most malignant cancers in the world. Cold micro plasma jet (CMPJ) Known as cold atmospheric jet microplasma, it has recently been introduced as an alternative way to overcome the challenges of finding an effective cancer treatment. Numerous studies have reported promising results, so our aim of this study was to investigate how this method affects cell death and its role on the expression of apoptotic genes in the MCF-7 cancer cell line. Methods: In this study, helium gas was used to generate plasma at room temperature in the form of point radiation at different times of 30, 60, 90 and 120 seconds and at different distances of 1 cm. Flow cytometry will be used to examine the extent of apoptosis and necrosis. Genes involved in apoptosis P53, P21, Bax and Bcl-2 were measured by real-time PCR. Results: Our Studie indicate that the mechanism of action of cold plasma on cancer cells is related to generation of reactive oxygen species with possible induction of the apoptosis pathway. The percentages of necrotic and late apoptotic cells following treatment with different times plasma (0, 30, 60, 90 and 120 s) were about 0.55 ± 0.06, 20.13 ± 0.01, 20.12 ± 0.03, 26.81 ± 0.04 and 17.51 ± 0.05. The mRNA expression of bcl-2 showed a decrease of 90s of plasma while the mRNA expression of p53, bax and caspase-8 genes increased compared to untreated cells. Conclusion: In general, research in the last decade has confirmed the ability of CMPJ as an effective anti-cancer tool. Therefore, it may be used to help treat cancer. However, its clinical application requires much further studies to determine the severity and duration of exposure to CMPJ for effective treatment based on the type of cancer.

Abstract P311: Blocking Succinate Release Enhances Mitochondrial Reactive Oxygen Species Generation And Worsens Ischemia-reperfusion Injury

Succinate is a metabolite that plays a central role in ischemia-reperfusion (IR) injury,which is relevant to myocardial infarction (heart attack) and stroke. Succinateaccumulates during ischemia and is rapidly consumed at reperfusion driving reactiveoxygen species (ROS) generation at complex-I (Cx-I) and III of the mitochondrial electrontransport chain. This ROS production triggers cell-death, leading to tissue necrosis.Although succinate oxidation has been extensively studied and exploited as a noveltherapeutic target, only 1/3 of the succinate accumulated in ischemia is oxidized atreperfusion, with the remaining 2/3 being released from the cell via monocarboxylatetransporter 1 (MCT1). Extracellular succinate is thought to be pro-inflammatory, and ithas been proposed that preventing succinate release may be therapeutically beneficial.To determine the impact of preventing succinate release on IR injury, we comparedfunctional recovery (i.e. rate x pressure product, RPP) and infarction (i.e. tissue necrosis)of Langendorff perfused mouse hearts treated with an MCT1 inhibitor, AR-C155858,versus vehicle control. This revealed that succinate retention worsens IR injury (i.e.increased infarction and decreased functional recovery) likely due to increased ROS. Totest this hypothesis, we utilized a Langendorff apparatus positioned within aspectrofluorimeter, which permits real-time fluorescence measurements in beatingmouse hearts. Using the mitochondria targeted superoxide probe, MitoSOX red tomeasure ROS production at reperfusion + AR-C155858, demonstrated that succinateretention leads to enhanced mitochondrial ROS generation at the onset of reperfusion.Overall, these results suggest that inhibiting succinate release in the context of IR injurymay not be a viable therapeutic approach, regardless of any downstream anti-inflammatory effects.

Abstract P346: Ferroptosis Is Critical In Diabetic Atherosclerosis Through Heme Oxygenase 1 Regulated Lipid Peroxidation Pathway

Introduction: The patients with diabetes dramatically accelerates atherosclerosis. Although improved diabetes treatment strategy has prevented the pathogenesis, diabetic atherosclerosis is still the main cause of morbidity and mortality. It is urgent to clarify its in-depth mechanism and identify new therapeutic targets for preventing diabetic atherosclerosis. Hypothesis: We assessed the hypothesis that ferroptosis and HMOX1 may be implicated in diabetes atherosclerosis through comprehensive in-depth bioinformatics analysis. Methods and Results: Analysis of Gene expression profiles (GSE30169 and GSE6584) provided a total of 91 differentially expressed genes (DEGs) including 68 up-regulated and 23 down-regulated genes (A). Ferroptosis, a type of programmed cell death dependent on iron, and HMOX1, a gene that encodes heme oxygenase, were identified as key factors in diabetes atherosclerosis (B). To explore the potential mechanism, we measured iron content and reactive oxygen species (ROS) generation and conducted a lipid peroxidation assay. Compared with animals on a normal diet (ND), ferroptosis and HMOX1 expression in ApoE -/- mice on high-fat diet (HFD) were significantly increased (C, n = 3). Ferroptosis inhibitor ferrostatin-1 (Fer-1) effectively attenuated the diabetic atherosclerosis area in HFD animals (D, n = 5, P < 0.01 vs HFD). In addition, Fer-1 restored the ferroptosis-related protein (GPX4 and SLC7A11) with high glucose high lipid (HGHL) treatment in Mouse arterial endothelial cells (MAECs) (E, n=6). More importantly, HMOX1 deficiency rebalanced Fe 2+ overload, decreased iron content, reduced ROS, and alleviated the lipid peroxidation, which led to the reduction of ferroptosis in diabetic endothelial cells (F). Conclusions: We demonstrate that ferroptosis is critical in diabetic AS progression and HMOX1 is an implicated factor responsible for ferroptosis in diabetic AS development, suggesting HMOX1 may serve as a potential therapeutic or drug development target.