Increased Lactate Dehydrogenase Activity Research Articles

Functional and biochemical characteristics of the muscle system in children with type I diabetes

Background. The purpose of the study was to determine possible markers of skeletal muscle damage in children with type 1 diabetes mellitus (T1DM) and their relationship with the features of disease course. Materials and methods. The observation group consisted of 98 children with type 1 diabetes mellitus: the first group included 22 people without disorders of the muscular system; the second — 42 patients with dynapenia; the third — 34 children with diabetic myopathy. Control group — 30 relatively healthy children. Assessment of the static endurance of skeletal muscles, determination of the level of creatine kinase, aspartate aminotransferase, alkaline phosphatase, lactate dehydrogenase, periostin and cardiotrophin-1 in blood serum were performed in all patients. Results. The conducted studies demonstrate that children with diabetes, regardless of the structural and functional state of their muscular system, have signs of skeletal muscle damage, which were most expressed in diabetic myopathy and progressed with maximal deterioration of glycemic control. It was found that the highest content of alkaline phosphatase was characteristic of children from group 1, while in patients with diabetic myopathy its serum content was not statistically different from that of controls. These disorders occurred against the background of changes in alkaline phosphatase activity, the level of which was highest in children from group 1, while in patients with diabetic myopathy, its serum content was not statistically different from that of controls. At the same time, during the course of diabetic myopathy in children with T1DM, there was an increase in lactate dehydrogenase activity by 1.2 times (p < 0.01) and cardiotrophin-1 by 300 times (p < 0.01) compared to the corresponding indicator of the control group. Serum periostin level was increased in all patients with T1DM. Its maximum values were determined in group 1, whose periostin concentration exceeded control indicators by 103 times (p < 0.01). With deterioration of skeletal muscle state, there was a gradual decrease in periostin serum level, but in patients with dynapenia, it was 35.5 times higher than in the control group (p < 0.05) and 19.2 times higher in those with diabetic myopathy (p < 0.05). Conclusions. The course of type 1 diabetes in children is accompanied by skeletal muscle damage, the first clinical sign of which is a decrease in the static muscle endurance against the background of worsening disease course. Alkaline phosphatase, lactate dehydrogenase, periostin, and cardiotrophin-1 are biochemical markers of skeletal muscle damage in children with type 1 diabetes. A common feature of the changes in the specified indicators is their increase; however, each clinical condition of the skeletal muscles corresponds to its own configuration of changes in the abovementioned markers.

Development of a novel bioengineered 3D brain-like tissue for studying primary blast-induced traumatic brain injury.

Primary blast injury is caused by the direct impact of an overpressurization wave on the body. Due to limitations of current models, we have developed a novel approach to study primary blast-induced traumatic brain injury. Specifically, we employ a bioengineered 3D brain-like human tissue culture system composed of collagen-infused silk protein donut-like hydrogels embedded with human IPSC-derived neurons, human astrocytes, and a human microglial cell line. We have utilized this system within an advanced blast simulator (ABS) to expose the 3D brain cultures to a blast wave that can be precisely controlled. These 3D cultures are enclosed in a 3D-printed surrogate skull-like material containing media which are then placed in a holder apparatus inside the ABS. This allows for exposure to the blast wave alone without any secondary injury occurring. We show that blast induces an increase in lactate dehydrogenase activity and glutamate release from the cultures, indicating cellular injury. Additionally, we observe a significant increase in axonal varicosities after blast. These varicosities can be stained with antibodies recognizing amyloid precursor protein. The presence of amyloid precursor protein deposits may indicate a blast-induced axonal transport deficit. After blast injury, we find a transient release of the known TBI biomarkers, UCHL1 and NF-H at 6 h and a delayed increase in S100B at 24 and 48 h. This in vitro model will enable us to gain a better understanding of clinically relevant pathological changes that occur following primary blast and can also be utilized for discovery and characterization of biomarkers.

Metabolic changes in elderly people with sarcopenia

Sarcopenia is a special condition that develops in older people. Sarcopenia represents a loss of muscle strength and mass in the elderly and is a common disease and is also associated with several adverse health effects. Due to the impact of sarcopenia on quality of life, disability and mortality, greater awareness is needed to correctly identify this condition and in particular its biological immunoendocrine markers. The purpose of the study was to study the characteristics of metabolism in sarcopenia in elderly people at the molecular, cellular and organizational levels. Material and methods. The study included 402 elderly people (199 men and 203 women, average age 68.9 ± 1.2 years), who are divided into three groups depending on the presence or absence of arterial hypertension and sarcopenia. The state of adipose and muscle tissue was studied using anthropometric measurements, bioimpedansometry and dynamometry. Results and discussion. The present study revealed a significant increase in energy exchange tension in patients with arterial hypertension, expressed in the increase in ADP content, but it was not enough to maintain the ATP/ADP ratio at the proper level. In patients with arterial hypertension and sarcopenia, a pronounced deficiency in ATP and ADP content, as well as ATP/ADP ratio was observed. The energy deficiency can also be explained by increase in lactate dehydrogenase activity in patients with arterial hypertension to compensate for energy deficiency by enhancing glycolysis processes. However, when sarcopenia was attached, this sanogenetic compensatory mechanism no longer worked. Conclusions. Dysfunction of adipose tissue in patients with arterial hypertension precedes the development of sarcopenia, while there is an increase in tension in energy exchange, expressed in elevation of ADP content, but when sarcopenia joins, there is a pronounced deficiency in both ATP and ADP content, as well as ATP/ADP ratio.

Effect of low-dose Piscirickettsia salmonis infection on haematological-biochemical blood parameters in Atlantic salmon (Salmo salar).

Piscirickettsia salmonis is the etiological agent of Piscirickettsiosis, a severe disease that affects Atlantic salmon (Salmo salar) farmed in Chile and many other areas (Norway, Scotland, Ireland, Canada and the USA). This study investigated the effects of low-dose P. salmonis infection (1 × 102 CFU/ml) on Atlantic salmon. In this study, we challenged fish with an isolated representative of the EM-90 genogroup via intraperitoneal injection for 42 days. Infected fish displayed decreased haematocrit and haemoglobin levels at day 13 post-infection, indicating erythropenia, haemolysis and haemodilution. Conversely, their white blood cell counts increased on days 13 and 21 post-infection. Additionally, their iron levels decreased from day 2 post-infection, indicating iron deficiency and an inability to retrieve stored iron before infection. Their magnesium levels also decreased at day 28 post-infection, possibly due to osmoregulatory problems. Also, we observed an increase in lactate dehydrogenase activity on days 5, 21, and 28 post-infection, suggesting early symptoms of hepatotoxicity. Later analyses determined a decrease in plasma glucose levels from day 2 post-infection. This may be attributed to the hypoxic conditions caused by P. salmonis, leading to an excess utilization of stored carbohydrates. Our results suggest that the blood parameters we studied are useful for monitoring the physiological status of Atlantic salmon infected with P. salmonis.

Oxidative stress as the trigger for menthol-induced developmental alterations in zebrafish (Danio rerio) model

Menthol is a natural origin organic compound obtained from the peppermint plant whose effects at the embryonic development level are not completely known. In this work, the effects of an acute exposure (96 h) to menthol were evaluated in zebrafish embryos based on the calculated lethal concentration (LC50, 27.5 mg L−1). The results showed a decreased hatching rate and heart rate as well as a notable increase in malformations and mortality. After menthol exposure, an increased production of reactive oxygen species (ROS) accompanied by an increase in the activity of antioxidant enzymes (superoxide dismutase, catalase, glutathione peroxidase and reductase) and a decrease in glutathione-S-transferase and protein carbonyls was detected. At the metabolic level, a decrease in ATPase activity and an increase in lactate dehydrogenase activity were perceived. Furthermore, mitochondrial membrane potential (ΔΨm) was increased after menthol exposure as well as the expression of Nrf2. At the behavioural level, shorter distance, speed, and percentage of active time were observed in menthol-treated larvae. In addition, an increase in the distance to the centre of the well and in the turn angle was observed supporting an anxiety-like behaviour induced by menthol exposure. Furthermore, there was a decrease in dopaminergic neurons and in the activity of the enzyme acetylcholinesterase (AChE) suggesting neurobehavioral impairment. Overall, while further studies are required, menthol compromised the development of zebrafish by mediating early development through the regulation of Nrf2 by mitochondrial ROS production with the consequent teratogenic outcomes.

Toxicity Assessment of Resveratrol Liposomes-in-Hydrogel Delivery System by EpiVaginalTM Tissue Model.

The natural polyphenol resveratrol (RES) has shown great potential as an antimicrobial, including against microbes associated with vaginal infections. To fully exploit the activities of RES, an all-natural ingredients formulation for RES delivery at vaginal site has been developed, namely liposomes loaded with RES, incorporated into a chitosan hydrogel as secondary vehicle. Although considered non-toxic and safe on their own, the compatibility of the final formulation must be evaluated for its biocompatibility and non-irritancy to the vaginal mucosa. As a preclinical safety assessment, the impact of RES formulation on the tissue viability, the effect on barrier function and cell monolayer integrity, and cytotoxicity were evaluated using the cell-based vaginal tissue model, the EpiVaginal™ tissue. RES liposomes-in-hydrogel formulations neither affected the mitochondrial activity, nor the integrity of the cell monolayer in RES concentration up to 60 µg/mL. Moreover, the barrier function was maintained to a greater extent by RES in formulation, emphasizing the benefits of the delivery system. Additionally, none of the tested formulations expressed an increase in lactate dehydrogenase activity compared to the non-treated tissues. The evaluation of the RES delivery system suggests that it is non-irritant and biocompatible with vaginal tissue in vitro in the RES concentrations considered as therapeutic.

CTRP12 alleviates cardiomyocyte ischemia‑reperfusion injury via regulation of KLF15.

Myocardial ischemia-reperfusion (I/R) serves a crucial role in myocardial infarction. C1q/TNF-related protein 12 (CTRP12) is a secretory protein involved in metabolism. It has been reported that CTRP12 participates in the regulation of numerous cardiovascular diseases. However, its role in myocardial I/R injury remains unclear. In the present study, the left anterior descending coronary artery in mice was ligated to establish a mouse I/R model. A myocardial hypoxia-reoxygenation (H/R) cell model was also established. Cardiomyocyte injury was evaluated using hematoxylin and eosin staining, Cell Counting Kit-8 and a lactate dehydrogenase (LDH) kit. The expression levels of CTRP12 and Krueppel-like factor 15 (KLF15) in murine myocardial tissues and H9c2 cells were determined using reverse transcription-quantitative PCR and western blotting, as KLF15 was previously reported to protect against I/R-induced cardiomyocyte damage. Furthermore, inflammatory factors TNF-α, IL-1β and IL-6 were analyzed using ELISA while apoptosis was assessed using TUNEL assays and western blotting. Moreover, the activity of the CTRP12 promoter was determined using a dual-luciferase reporter assay. The results demonstrated that I/R surgery markedly exacerbated myocardial tissue damage, whereas H/R treatment significantly reduced cell viability and significantly increased LDH activity as well as the release of inflammatory factors and apoptosis. I/R and H/R induction significantly reduced the expression levels of CTRP12 and KLF15. CTRP12 overexpression significantly alleviated H/R-induced cell injury and significantly inhibited inflammation and apoptosis. Further analysis demonstrated that KLF15 could significantly promote the activity of the CTRP12 promoter. However, following CTRP12 knockdown, KLF15 overexpression exacerbated cell injury, inflammation and apoptosis. In conclusion, the present study demonstrated that CTRP12 may mitigate inflammation and apoptosis in H/R-induced cardiomyocytes, possibly via the regulation of KLF15, which provided a theoretical basis for the potential treatment of I/R-induced myocardial infarction.

Fumonisin B1 Induces Immunotoxicity and Apoptosis of Chicken Splenic Lymphocytes

Fumonisin B1 (FB1), produced by Fusarium, is among the most abundant and toxic mycotoxin contaminations in feed, causing damages to the health of livestock. However, the mechanisms of FB1 toxicity in chickens are less understood. As splenic lymphocytes play important roles in the immune system, the aim of this study was to investigate the immunotoxic effects and mechanisms of FB1 on chicken splenic lymphocytes. In the present study, the chicken primary splenic lymphocytes were harvested and treated with 0, 2.5, 5, 10, 20 and 40 μg/mL FB1. Then, the cell proliferation, damage, ultrastructure, inflammation and apoptosis were evaluated. Results showed that the proliferation rate of splenic lymphocytes was decreased by FB1 treatments. The activity of lactate dehydrogenase (LDH) was increased by FB1 treatments in a dose-dependent manner, implying the induction of cell damage. Consistently, the ultrastructure of splenic lymphocytes showed that FB1 at all the tested concentrations caused cell structure alterations, including nuclear vacuolation, mitochondrial swelling and mitochondrial crest fracture. Besides, immunosuppressive effects of FB1 were observed by the decreased concentrations of interleukin-2 (IL-2), IL-4, IL-12 and interferon-γ (IFN-γ) in the cell culture supernatant. Furthermore, apoptosis was observed in FB1-treated cells by flow cytometry. The mRNA expressions of apoptosis-related genes showed that the expression of Bcl-2 was decreased, while the expressions of the P53, Bax, Bak-1, and Caspase-3 were increased with FB1 treatment. Similar results were found in the concentrations of apoptosis-related proteins in the cell supernatant by ELISA assay. Moreover, regression analysis indicated that increasing FB1 concentration increased LDH activity, concentrations of Bax, Bak-1 and mRNA expression of Bak-1 linearly, increased M1 area percentage quadratically, decreased concentration of IFN-γ, mRNA expression of Bcl-2 linearly, and decreased concentrations of IL-2 and IL-4 quadratically. Besides, regression analysis also showed reciprocal relationships between IL-12 concentration, Caspase-3 mRNA expression and increasing FB1 concentration. The increasing FB1 concentration could decrease IL-12 concentration and increase Caspase-3 mRNA expression. Altogether, this study reported that FB1 induced the immunotoxicity of chicken splenic lymphocytes and caused splenic lymphocytes apoptosis by the Bcl-2 family-mediated mitochondrial pathway of caspase activation.

MicroRNA-503 Exacerbates Myocardial Ischemia/Reperfusion Injury via Inhibiting PI3K/Akt- and STAT3-Dependent Prosurvival Signaling Pathways.

Acute myocardial infarction is a leading cause of death worldwide, while restoration of blood flow to previously ischemic myocardium may lead to ischemia/reperfusion (I/R) injury. Accumulated evidence shows that microRNAs play important roles in cardiovascular diseases. However, the potential role of microRNA-503 (miR-503) in myocardial I/R injury is little known. Thus, this study is aimed at determining whether and how miR-503 affects myocardial I/R injury in vivo and in vitro. A mouse model of myocardial I/R injury and H9c2 cell model of hypoxia/reoxygenation (H/R) injury were established. The postischemic cardiac miR-503 was downregulated in vivo and in vitro. Mechanistically, PI3K p85 and Bcl-2 are miR-503 targets. The post-ischemic cardiac PI3K p85 protein level was decreased in vivo. Agomir-503 treatment exacerbated H/R-induced injuries manifested as decreased cell viability, increased lactate dehydrogenase activity, and cell apoptosis. Agomir-503 treatment reduced cell viability under normoxia as well and reduced both PI3K p85 and Bcl-2 protein levels under either normoxia or H/R condition. It reduced phosphorylation of Stat3 (p-Stat3-Y705) and Akt (T450) in cells subjected to H/R. In contrast, Antagomir-503 treatment attenuated H/R injury and increased p-Stat3 (Y705) under normoxia and increased p-Akt (T450) under either normoxia or H/R condition. It is concluded that miR-503 exacerbated I/R injury via inactivation of PI3K/Akt and STAT3 pathways and may become a therapeutic target in preventing myocardial I/R injury.

Impact of recentrifugation of blood collection tubes on chemistry and immunochemistry analytes after 24 and 72 hours of refrigerated storage on the Roche Cobas 8000 platform

BackgroundWe discovered that blood collection tubes (BCTs) were inadvertently recentrifuged due to improper placement on our automated preanalytical system. This study was undertaken to determine the impact of recentrifugation of blood specimens collected in serum separator (SSTs) and plasma separator (PSTs) tubes after refrigerated storage for 24 and 72 h on the concentrations of chemistry and immunochemistry analytes. MethodsBlood was collected from 20 volunteers in SSTs and PSTs, centrifuged, and 36 chemistry and 14 immunochemistry analytes were measured at baseline in single-centrifuged tubes on a Roche Cobas 8000 chemistry platform. After baseline testing, the BCTs were refrigerated for 24 or 72 h, recentrifuged and retested. The results were compared to the single-centrifuged tubes for statistical significance. ResultsRecentrifugation of BCTs after 24 or 72 h of refrigerated storage showed statistically significant increases in lactate dehydrogenase activity and potassium concentration and statistically significant decreases in glucose (except in SSTs after 24 h of refrigerated storage) and CO2 concentration, but no significant differences in immunochemistry analyte concentrations. ConclusionIt may be safe to report most routine chemistry and immunochemistry analyte concentrations from recentrifuged SSTs and PSTs on the Roche Cobas 8000, which may save time and costs associated with recollection and retesting.

Tolerance of juvenile Peruvian rock seabass (Paralabrax humeralis Valenciennes, 1828) and Peruvian grunt (Anisotremus scapularis Tschudi, 1846) to low-oxygen conditions.

Hypoxia is currently one of the greatest threats to coastal ecosystems worldwide, generating massive mortality of marine organisms, loss of benthic ecosystems and a decrease in fishery production. We evaluated and compared the tolerance to hypoxia of two species from different habitats of the Peruvian coast, the Peruvian rock seabass Paralabrax humeralis and the Peruvian grunt Anisotremus scapularis. The effect of hypoxia was measured as a function of the exposure time (progressive and chronic) on the behavioural and physiological responses of the two species, as well as on the enzymatic activity associated with the oxidative stress response of lactate dehydrogenase (LDH), superoxide dismutase (SOD) and alkaline phosphatase (AKP). The ventilatory frequency was measured at two different temperatures (16 and 22°C) under progressive hypoxia conditions to determine the ventilatory critical point (Vcp). A. scapularis showed a higher Vcp than P. humeralis, which was positively affected by temperature. The median lethal time of A. scapularis was 36 min at 60% of oxygen saturation, while P. humeralis showed no mortality after 31 days of exposure at 5% oxygen saturation. Different enzymatic activity (P < 0.05) between species under hypoxia was recorded, in SOD (gill and muscle) and AKP (blood). A general tendency, under hypoxia, to slightly increase LDH activity (except for blood in A. scapularis, P< 0.05) and SOD activity (mainly in muscle of A. scapularis, P< 0.05), and decrease AKP activity (mainly in liver of P. humeralis, P< 0.05) was observed. The response of P. humeralis to hypoxia goes through a reduction in activity and metabolism, so this species can be considered hypoxia-tolerant, allowing it to face hypoxia events during prolonged periods. On the other hand, A. scapularis response to hypoxia prioritizes avoidance mechanisms and, together with other adaptations, makes it especially vulnerable to hypoxia and able to be considered hypoxia-intolerant.

Subchronic Toxicity of Ivermectin and Butaphosphan in Layer Chickens

The development of new veterinary drugs to treat and prevent poultry parasitic infections, as well as the study of their safety is a hot topic for modern parasitology. The purpose of this research was to study the subchronic toxicity of the ivermectin and butaphosphan-based drugs at a therapeutic and threefold therapeutic dose during a seven-day oral administration to the Hisex White chickens. The provisional name of the drug is Iverbutan. The chickens from the first experimental group were given the drug at a threefold therapeutic dose of 3 mL of the drug per one liter of drinking water. The chickens from the second experimental group were given the drug at a therapeutic dose of 1 mL of Iverbutan per one liter of drinking water. The chickens from the control group received water without the drug. The chickens were weighed, and then the body temperature and blood samples from the axillary vein were measured on days 1, 8, and 17 of the experiment before the morning feeding. On day 8 of the study, the chickens from the first experimental group showed a 7.4% decrease in body weight and increase in body temperature by 0.8%, an increase in alanine aminotransferase activity by 2.1 times, aspartate aminotransferase activity by 1.6 times, and bile acids by 1.4 times. Moreover, there was a 4.6% decrease in glucose concentration, a 3.5% increase in lactate dehydrogenase activity, a 7.3% decrease in triglycerides, as well as a decrease in hemoglobin by 3.2% and erythrocytes by 10.6% in the first experimental group, compared to the control group. On day 17 of the experiment, the above blood parameters in the chickens from the first experimental group did not significantly differ from the control group, indicating the reversibility of the hepatotoxic effect. In this regard, a three-fold therapeutic dose can be considered a threshold. The chickens from the second experimental group showed no changes in their physiological status as compared to the control. Thus, the study results confirm the safety of the drug in the recommended dosage regimen.

Effects of N Ethyl N′ nitrosourea in mice brain in time fashion

Antioxidant enzyme plays a pivotal role in preventing oxidative stress. Chemical toxicants often exert adverse effects in the biological system by breaking the antioxidant response system leading to severe oxidative stress. N-Ethyl-N-Nitrosourea (ENU) is a DNA alkylating agent and is carcinogenic and neurotoxic in animals. ENU in the nervous system cause persistent alkylation of DNA within the neurons altering the normal functional activities of the brain. Prenatal exposure to ENU in rodents that generate malignant gliomas in the brain induces functional changes and apoptotic death in neuronal progenitors in the brain's subventricular zone (SVZ). However, as chemical toxicants, their role in the induction of oxidative stress in brain tissue and subsequent change in the brain were less studied. In this study, we have investigated the response antioxidant system and induction of oxidative stress in the Balb/c mice brain at various time points after ENU exposure. Exposures to ENU were found to raise the oxidative stress and lipid peroxidation in the brain of mice with subsequent reduction in antioxidant enzyme activity. A significant increase in lactate dehydrogenase activity in the serum and brain homogenate was recorded at 16th weeks after ENU injection, which indicates metabolic alteration due to damage in the brain and has been validated by histological damage in the cerebral cortex and abnormal neurologic behavior in animals administered with ENU.

Platycodin D regulates high glucose-induced ferroptosis of HK-2 cells through glutathione peroxidase 4 (GPX4)

ABSTRACT Diabetic nephropathy (DN) is associated with inflammation. Platycodin D (PD) demonstrates anti-inflammatory activity. However, whether PD affects DN remains to be explored. Here, we aimed to discuss the role of PD in DN and its underlying mechanisms. High glucose (HG)-induced HK-2 cells were treated with PD, and cell viability was assessed using the Thiazolyl Blue Tetrazolium Bromide (MTT) assay. Ferroptosis-related factors such as lactate dehydrogenase (LDH) activity, lipid reactive oxygen species (ROS), iron (Fe2+) level, GSH level, and malondialdehyde (MDA) level were evaluated. Cell death was evaluated using the TUNEL assay. GPX4 expression was evaluated using Quantitative Real-time PCR (qRT-PCR) and Western blotting analysis. The results indicated that HG increased LDH activity, lipid ROS production, Fe2+ levels, and MDA levels and decreased GSH levels, suggesting that the HG condition induced ferroptosis. PD treatment inhibited ferroptosis in HG-induced cells, downregulated ACSL4 and TFR1 expression, and upregulated FTH-1 and SLC7A11 expression. PD reversed the effects of HG condition on cell death. Moreover, GPX4 expression was downregulated in HG-stimulated cells. Furthermore, we substantiated that PD suppressed ferroptosis by modulating GPX4 expression. In conclusion, PD inhibited ferroptosis in HG-induced HK-2 cells by upregulating GPX4 expression, suggesting that PD may be an effective drug for the clinical treatment of DN.

Induction of apoptosis and suppression of Ras gene expression in MCF human breast cancer cells.

Breast cancer is the leading invasive cancer in women globally. This study aimed at evaluating the anti-apoptotic activity of p-Coumaric acid (PCA) on MCF-7 breast cancer cell line. Experiments were conducted in which the MCF-7 cell line was treated with PCA. which showed decreased cell viability, increased lactate dehydrogenase activity, and caspase-3 activation. The results were evaluated with real-time polymerase chain reaction which revealed that PCA reduced the amount of H-Ras and K-Ras transcript in MCF-7 breast cancer cells. In the presence of PCA there was a significant increase in the levels of mRNA gene Bax and late apoptotic cells which was dose dependent. It also retarded the relative expression of antiapoptotic gene, Bcl2 in treated cells. The results suggest that PCA exhibits anti-cancer properties against MCF-7 cells. PCA inhibited the growth of MCF7 cell. The optimum concentration of PCA was 75-150 mM. PCA can inhibit the growth of MCF-7 cells by reducing Ras expression and inducing cell apoptosis. Our results suggest that PCA could prove valuable in the search for possible inhibitors of Ras oncogene functionality and gain further support for its potential utilization in the treatment of patients with breast cancer. PCA is safe and could complement current treatments employed for the disease.

Apigenin induces oxidative stress in mouse Sertoli TM4 cells.

Background and Aim:Apigenin (API) is an estrogenic compound found in many plants. Sertoli cells reside in the testis and are a key target of environmental toxicants. This study aimed to examine the cytotoxicity, especially oxidative stress of API in mouse Sertoli TM4 cells.Materials and Methods:Mouse Sertoli TM4 cells were treated with 50 and 100 μM API for 48 h. Cell viability, lactate dehydrogenase (LDH) activities, glutathione reductase (GR) activities, production of reactive oxygen species (ROS), and malondialdehyde (MDA) levels were evaluated using various assays.Results:Treatment with API at both 50 and 100 μM decreased viability and GR activity but increased LDH activity, ROS production, and MDA levels in mouse Sertoli TM4 cells.Conclusion:Exposure to API induced oxidative stress in mouse Sertoli TM4 cells.

Acute Exposure to the Penconazole-Containing Fungicide Topas Induces Metabolic Stress in Goldfish.

Triazole fungicides are widely used in agriculture that leads to pollution of freshwater ecosystems. The mechanisms of toxicity to fish by the triazole fungicide Topas that contains penconazole (1-[2-(2,4-dichlorophenyl)pentyl]-1H-1,2,4-triazole) have not been studied. The present study aimed to evaluate the effect of goldfish exposure for 96 h to the fungicide Topas at concentrations of 1.5, 15, or 25 mg/L on the plasma and liver biochemical parameters and blood hematological profile. Goldfish exposure to Topas decreased alanine and aspartate transaminase activity and increased lactate dehydrogenase activity in the liver. Plasma lactate dehydrogenase and alanine transaminase activities were elevated in fungicide-treated fish. Topas exposure also enhanced plasma glucose and triacylglycerol concentrations. In the liver, fungicide treatment decreased levels of glucose but elevated triacylglycerols, glycogen, and protein. The results indicate that acute exposure of goldfish to Topas induced strong metabolic perturbations and disruptions of metabolic parameters, suggesting that these could be used to assess sublethal or acute toxic effects of pesticides on aquatic species.

Autophagy and Apoptosis Induced in U87 MG Glioblastoma Cells by Hypericin-Mediated Photodynamic Therapy Can Be Photobiomodulated with 808 nm Light

Glioblastoma is one of the most aggressive types of tumors. Although few treatment options are currently available, new modalities are needed to improve prognosis. In this context, photodynamic therapy (PDT) is a promising adjuvant treatment modality. In the present work, hypericin-mediated PDT (hypericin-PDT, 2 J/cm2) of U87 MG cells is combined with (2 min, 15 mW/cm2 at 808 nm) photobiomodulation (PBM). We observed that PBM stimulates autophagy, which, in combination with PDT, increases the treatment efficacy and leads to apoptosis. Confocal fluorescence microscopy, cytotoxicity assays and Western blot were used to monitor apoptotic and autophagic processes in these cells. Destabilization of lysosomes, mitochondria and the Golgi apparatus led to an increase in lactate dehydrogenase activity, oxidative stress levels, LC3-II, and caspase-3, as well as a decrease of the PKCα and STAT3 protein levels in response to hypericin-PDT subcellular concentration in U87 MG cells. Our results indicate that therapeutic hypericin concentrations can be reduced when PDT is combined with PBM. This will likely allow to reduce the damage induced in surrounding healthy tissues when PBM-hypericin-PDT is used for in vivo tumor treatments.

Lactate dehydrogenase activity staining demonstrates time-dependent immune cell infiltration in human ex-vivo burn-injured skin

Burn injuries constitute one of the most serious accidental injuries. Increased metabolic rate is a hallmark feature of burn injury. Visualising lactate dehydrogenase (LDH) activity has been previously used to identify metabolic activity differences, hence cell viability and burn depth in burn skin. LDH activity was visualised in injured and uninjured skin from 38 sub-acute burn patients. LDH activity aided the identification of spatially correlating immunocompetent cells in a sub-group of six patients. Desorption Electrospray Ionisation Mass Spectrometry Imaging (DESI MSI) was used to describe relative lactate and pyruvate abundance in burned and uninjured tissue. LDH activity was significantly increased in the middle and deep regions of burnt skin compared with superficial areas in burnt skin and uninjured tissue and positively correlated with post-burn time. Regions of increased LDH activity showed high pyruvate and low lactate abundance when examined with DESI-MSI. Areas of increased LDH activity exhibited cellular infiltration, including CD3 + and CD4 + T-lymphocytes and CD68 + macrophages. Our data demonstrate a steady increase in functional LDH activity in sub-acute burn wounds linked to cellular infiltration. The cell types associated are related to tissue restructuring and inflammation. This region in burn wounds is likely the focus of dysregulated inflammation and hypermetabolism.

Ameliorative effect of piracetam on emamectin benzoate induced perturbations in the activity of lactate dehydrogenase in murine system

Emamectin benzoate (4´-deoxy-4´-epimethylaminoavermectin B1) is a macrocyclic lactone developed for the control of insect pests in edible plant crops. The current study was taken to assess the possible ameliorative effect of piracetam (P) on emamectin benzoate (EMB) induced alterations in energy metabolism in liver, kidney, testis, heart, cerebral cortex, cerebellum and muscle of rats. Rats of normal control group (N) received water and normal diet ad libitum and (EMB as well as EMB+P) group received 5 mg/kg bw emamectin benzoate via drinking water for 4 weeks. After four weeks of EMB treatment, rats of (EMB+P) received piracetam (500 mg/kg bw, iv) for 15 days. The results demonstrate that emamectin benzoate caused a significant increase in lactate dehydrogenase activity and the level of lactate indicating organ damage in treated animals. Piracetam, the so-called ‘nootropic’ drug is mainly used to treat cognitive impairment in aging, brain injuries, as well as dementia. In the present study piracetam treatment caused a significant inhibition of lactate dehydrogenase activity in liver, kidney, testis, heart, cerebral cortex, cerebellum and muscle of rats. The amelioration of toxic effects by piracetam treatment in EMB treated rats indicate that this drug has the potential to mitigate emamectin benzoate induced toxicity in mammalian systems.