Lactate Dehydrogenase Activity Research Articles

Bioaccumulation, transformation and toxicity of imidacloprid and dinotefuran in Eisenia fetida under single and binary exposure scenarios

Earthworms (Eisenia fetida) were exposed to individual and binary mixture of imidacloprid (IMI) and dinotefuran (DIN) at 0.05 and 0.5 mg/kg for 28 days to investigate their bioaccumulation, transformation and toxicity. IMI was more easily absorbed by earthworms than DIN, and worms didn’t accumulate or generate toxic metabolites. The obvious accumulation of neonicotinoids during later period caused significant neural dysfunction, especially when exposed to high-concentration IMI. Meanwhile, oxidative stress indicated by decreased SOD/CAT activity (33.2 %-68.1 %) and increased MDA (38.4 %-55.0 %) was induced by binary exposure with high-concentration IMI. By contrast, coelomocytes responded earlier and more strongly than oxidative responses. Coelomocytes’ viability and mitochondrial membrane potential were inhibited (23.6 %-91.7 %) mainly by IMI and binary exposure. Coelomocytes’ lactate dehydrogenase activity exerted a fluctuating pattern, suggesting irregular disturbance on cellular functions. This study highlights the role of coelomocytes and the need to consider binary/multiple scenarios and transformation of neonicotinoids in their risk assessment to earthworms.

Adaptive capability of slow-growing backyard poultry as indicated by physiological and molecular responses in a hot and humid coastal climate

Assessing the adaptability of slow-growing rural chickens for improving thermotolerance to suit the global climate change is a major research need. This work was aimed to evaluate the adaptability of CARI-Debendra chickens and to identify the polymorphism as well as expression profiling of thermotolerant genes (HSP70 and GRP78) under prevailing temperature-humidity indices and thermal stress in a coastal environment. One hundred sixty straight run chicks were reared at THI≥75 (control) and THI>80 under coastal climate till 12 weeks. Polymorphism of HSP70 and GRP78 candidate genes were explored using restriction enzymes TaqI and HaeIII to identify possible thermotolerance markers. Expression profiling of both the genes in liver, intestine and pectoralis muscle was determined through quantitative real-time PCR. Rectal and body surface temperature recorded in the neck and back showed significant differences (P < 0.01) with higher temperature in THI>80 group. Comparatively lower live weights (P < 0.05) and poor FCR were recorded in THI>80 group. The villi height in all intestinal segments was significantly lower (P < 0.01), but deeper crypt depth was observed in THI>80 than control group. A lowest thymus weight (P < 0.05) was noted with no significant differences in immune response in treatment group. Serum levels of cholesterol, activities of lactate dehydrogenase, creatinine kinase and concentration of potassium, sodium and thyroxine hormone were not different between the 2 groups. The concentration of triiodothyronine and chloride ion was lower in THI>80 group indicating adaptive changes for thermoregulation. HSP70 gene expressions in the three tissues were differentially increased (P < 0.01) by temperature-humidity indices, but the expression of GRP78 was not different between the 2 groups. The results concluded that the environmental factors interact with genetics on adaptability towards thermotolerance in slow-growing chickens.

Chemotherapy-induced peripheral neuropathy in the neuronal cells model

This research aims to investigate the impacts of quercetin and low-level laser therapy (LLLT) on undifferentiated and nerve growth factor-differentiated PC12 cells experiencing cisplatin-induced peripheral neuropathy. PC12 cells treated with cisplatin were simultaneously exposed to quercetin and LLLT. We evaluated the effects of quercetin and LLLT on the expression of GAP-43 and Synapsin I genes through real-time PCR, cell viability using MTT assay, apoptosis induction via Annexin and dead assay, mitochondrial potential alterations using a mitopotential assay, and lactate dehydrogenase activity analysis in cells. Our findings revealed that cisplatin led to increased apoptosis, mitochondrial dysfunction, and LDH activity, while reducing cell viability and inhibiting the expression of GAP-43 and Synapsin I genes in both undifferentiated and differentiated PC12 cells. However, the application of quercetin and LLLT resulted in decreased apoptosis, mitochondrial potential alterations, and lactate dehydrogenase activity. We propose further investigation into the in vivo effects of quercetin and low-level laser therapy on cisplatin-induced peripheral neuropathy, as well as exploring the molecular relationship between quercetin and low-level laser therapy.

Comparative Study of the Protective Effects of Citral, Thymoquinone, and Silymarin on Methotrexate-induced Cardiotoxicity in Rats.

Methotrexate (MTX), an immunosuppressant and anti-cancer medication, can harm the heart. The goal of the current investigation was to assess the cardiotoxicity caused by MTX and the potential cardioprotective properties of silymarin, citral, and thymoquinone as antioxidants. Forty-eight rats were divided into six groups, which included control, MTX, cosolvent, citral, thymoquinone, and silymarin groups. At the end of the study, the rats were anesthetized (ketamine and xylazine) and killed using CO2. Their blood samples were collected to measure the enzymatic activities of creatine kinase-myoglobin binding (CK-MB), creatine phosphokinase (CPK), and lactate dehydrogenase (LDH). Also, the heart tissue was sampled to determine the antioxidant capacity and examine the histopathology. The findings revealed that the activity of CPK, CK-MB, and LDH enzymes significantly reduced in the thymoquinone treatment group compared to the MTX group (p < 0.05). On the other hand, total antioxidant capacity was significantly increased in the thymoquinone group compared to the MTX group (p < 0.05). The pathological modifications (i.e. severe congestion, edema fluid, the presence of inflammatory cells around the blood vessels, mild to moderate hemorrhaging between cardiac muscle fibers) were seen in the MTX group. The treatment groups, particularly thymoquinone, did not experience any appreciable pathological changes. The thymoquinone was found to have the strongest protective effect against the heart damage caused by MTX.

Effects of season on metabolic profile of Holstein Friesian cows in postpartum period

The aim of the present study was to determine the metabolic profile of Holstein-Friesian cows in the postpartum period, as well as the effect of season on metabolic profile. The postpartum period is essential in the reproductive life of high yielding dairy cows because of its impact on future gravidity. This study included 60 cows up to 15 days after parturition, aged 2-8 years (the largest number of cows was between 3 and 5 years old) with no apparent clinical problems. Cows were sampled in summer season (n=30) and winter season (n=30). Parameters of metabolic profile were determined as follows: glucose, albumin, total protein, cholesterol, bilirubin, alanine aminotransferase (ALT), aspartate aminotransferase (AST), lactate dehydrogenase (LDH), urea, calcium, phosphorus and magnesium. Statistical differences were considered significant at the p&lt;0.05. Present research showed that all investigated parameters were within a reference range for cattle. Impact of season sampling was determined for glucose, albumin, total protein, cholesterol and phosphorus, while bilirubin, calcium, magnesium, urea as well as activities of ALT, AST and LDH were unaffected by the season of sampling. In conclusion, metabolic status is affected by the season and examination during the postpartum period can provide valuable information of cows' health status, in order to diagnose and moreover prevent postpartum diseases.

TRIM27 revealing by tumor educated platelet RNA-sequencing, as a potential biomarker for malignant ground-glass opacities diagnosis mediates glycolysis of non-small cell lung cancer cells partially through HOXM1.

Efficient ground-glass opacities (GGOs) diagnosis is challenging. A diagnostic method distinguishing malignant from benign GGOs is warranted. In this study, we sought to construct a noninvasive method based on tumor educated platelet (TEP) RNA profiles for malignant GGOs diagnosis and explore the molecular mechanism of the potential biomarker for the first time. Based on TEP RNA-sequencing (TEP RNA-seq) in benign and malignant GGOs, a classification model was constructed using differentially expressed genes (DEGs) and was used to evaluate diagnostic performance. High-throughput quantitative polymerase chain reaction (HT-qPCR) verified 23 genes selected from the top 60 DEGs between benign and malignant GGOs. The correlation between 17 verified DEGs and 22 key glycolytic genes was analyzed. Tripartite motif-containing 27 (TRIM27) overexpressing and knockdown (KD) cell models were constructed using A549 and PC-9 cells, respectively in which cell growth, apoptosis, migration and invasion were evaluated. The protein levels of HK-1/2, PKM1/2, LDHA and GLUT1 were evaluated by western blot. Glycolysis was evaluated through adenosine triphosphate (ATP), reactive oxygen species (ROS), lactate acid (LD) production, glucose uptake, and lactate dehydrogenase (LDH) activity assays. RNA-seq was performed in loss-of TRIM27-KD PC-9 cells to clarify the downstream factors of TRIM27 which was verified using western blot and immunofluorescence double staining. In 81 samples, the 1,647-DEG-based classification model exhibited area under the curve (AUC), sensitivity, and specificity values of 0.99 [95% confidence interval (CI): 0.972-1.000], 100%, and 91%, respectively, while the top 60-DEG-based classification model exhibited AUC, sensitivity, and specificity values of 0.986 (95% CI: 0.962-1.000), 98%, and 91%, respectively. TRIM27 achieved AUC of 0.87 in the diagnosis of malignant GGOs, with 83.93% sensitivity, 78.79% specificity, 81.15% accuracy, 77.05% positive predictive value (PPV) and 85.25% negative predictive value (NPV). TRIM27 was highly expressed in non-small cell lung cancer (NSCLC) cells, and accelerated cell migration and invasion. In addition, TRIM27 was found to promote glycolysis in NSCLC cells partially through HMOX1 which was negatively correlated with TRIM27. We constructed a novel TEP RNA-seq based classifier for malignant GGOs diagnosis. TRIM27, an important target discovered, could accelerate migration, invasion and regulate glycolysis partially through HMOX1 in NSCLC cells, thus providing scientific support for TRIM27 as a diagnostic biomarker for malignant GGO diagnosis.

The Antioxidant and Anti-Fatigue Effects of Rare Ginsenosides and γ-Aminobutyric Acid in Fermented Ginseng and Germinated Brown Rice Puree.

γ-aminobutyric acid (GABA) and rare ginsenosides are good antioxidant and anti-fatigue active components that can be enriched via probiotic fermentation. In this study, ginseng and germinated brown rice were used as raw materials to produce six fermented purees using fermentation and non-fermentation technology. We tested the chemical composition of the purees and found that the content of GABA and rare ginsenoside (Rh4, Rg3, and CK) in the puree made of ginseng and germinated brown rice (FGB) increased significantly after fermentation. The antioxidant activity of the six purees was determined using cell-free experiments, and it was found that FGB had better ferric-ion-reducing antioxidant power (FRAP) and 1,1-diphenyl-2-picryl-hydrazyl (DPPH) free radical scavenging rates, exhibiting better antioxidant effects. We then evaluated the antioxidant effect of FGB in HepG2 cells induced by H2O2 and found that FGB can reduce the generation of reactive oxygen species (ROS) in HepG2 cells and increase the membrane potential level, thereby improving oxidative damage in these cells. In vivo experiments also showed that FGB has good antioxidant and anti-fatigue activities, which can prolong the exhaustive swimming time of mice and reduce the accumulation of metabolites, and is accompanied by a corresponding increase in liver glycogen and muscle glycogen levels as well as superoxide dismutase and lactate dehydrogenase activities. Finally, we believe that the substances with good antioxidant and anti-fatigue activity found in FGB are derived from co-fermented enriched GABA and rare ginsenosides.

Study on sperm cryopreservation of hybrid fish derived from Carassius cuvieri (♀) × Carassius auratus red var (♂)

Cryopreservation of sperm is an effective method for conserving germplasm resources in fish genetic breeding. The high-quality hybrid fish (WR) derived from white crucian carp (Carassius cuvieri, WCC, ♀) and red crucian carp (C. auratus red var., RCC, ♂), possesses valuable traits such as high survival rates, strong resistance, and rapid growth, representing an important germplasm resources of crucian carp. This study compared the effects of different antifreeze solutions on sperm viability among three varieties (WR, WCC, and RCC) and examined changes in enzyme activity, fertilization rates, and hatching rates after cryopreservation, aiming to enhance the cryogenic sperm cryopreservation technique in hybrid fish and investigate the mechanisms underlying spermatozoa damage caused by cryopreservation. The results showed that the antifreeze combination of D14 with 15 % dimethyl sulfoxide (DMSO) had the best effect in preserving the sperm of WR and WCC, while D20 with 10 % DMSO was the optimal combination for RCC sperm. After ultra-low temperature preservation, the longevity, fertilization, and hatching rates of frozen sperm were significantly lower (P < 0.05) compared to fresh sperm. The enzyme activities of superoxide dismutase (SOD), lactate dehydrogenase (LDH), and creatine kinase (CK) were significantly decreased (P < 0.05) in spermatozoa, whereas they showed a significant increase (P < 0.05) in sperm plasma. Succinate dehydrogenase (SDH) activity was significantly reduced (P < 0.05) in frozen spermatozoa of RCC compared to fresh spermatozoa, and exhibited lower activity in frozen spermatozoa of WCC and WR. Additionally, SDH activity was significantly elevated (P < 0.05) in frozen sperm plasma, and glutathione reductase (GR) activity was significantly lower (P < 0.05) in both frozen sperm plasma and spermatozoa across all three species. The study demonstrated that cryopreservation had a significant effect on the enzyme activities of spermatozoa and sperm plasma in all three species. These findings provide important technical support for the conservation of high-quality fish germplasm resources, particularly for novel varieties resulting from distant hybridization in fish.

Dynamic changes of protein lactylation and their correlations with the glycolytic process during the postmortem acidification of broiler breast

This experiment aimed to reveal the dynamic changes of protein post-translational lactylation modifications and their correlations with the glycolytic process in broiler breast muscle within 48 h of postmortem acidification. The experiment involved 12 male AA broilers, 42 days old, with similar body weights (2.8 ± 0.05 kg). The breast fillets (Pectoralis major) were collected after slaughter, and samples were taken at various time points: 0, 15 min, 30 min, 45 min, 60 min, 2 h, 4 h, 6 h, 8 h, 12 h, 18 h, 24 h, 36 h, and 48 h postmortem. The results showed that the rate of glycogen decline in the muscle was highest at 45 min postmortem, and glycogen levels tended to stabilize at 8 h postmortem. The lactate content in the breast reached its highest level at 4 h postmortem and began to decrease, stabilizing at 24 h postmortem. Additionally, the glycolytic potential increased gradually in the first 4 h postmortem, decreased rapidly from 4 to 8 h. Similarly, lactylation modification levels were highest at 8 h postmortem, but stabilized at 12 h postmortem. During this process, the protein expression of the enzymatic lactylation modifier p300 showed no significant difference, while the content of the nonenzymatic lactylation substrate lactoylglutathione significantly decreased at 8 h and 24 h postmortem. Correlation analysis found that lactylation levels were negatively correlated with glycogen content, glucose content, glycolytic potential, and pH value, while positively correlated with lactate content. Besides, there was a positive correlation between lactylation levels and the protein expression of hexokinase, phosphoglycerate kinase 2, phosphoglucomutase 1, and triosephosphate isomerase. Additionally, lactylation levels were positively correlated with the activities of lactate dehydrogenase and phosphofructokinase. In summary, our experiment elucidated the dynamic changes in the entire glycolytic pathway in broiler pectoral muscle during acidification. During this process, lactylation modifications may participate in the glycolysis process by regulating the protein expression and activity of glycolytic enzymes.

Cinnamaldehyde Alleviates Alveolar Epithelial Cell Injury in ALI by Inhibiting the CaMKII Pathway.

Alveolar epithelial cell injury plays a key role in acute lung injury (ALI) and is a vital determinant of its severity. Here, we aimed to assess the protective effects of cinnamaldehyde (CA) on lipopolysaccharide (LPS)-induced A549 cells and elucidate the underlying mechanisms. A549 cells were stimulated with 1 μg/mL LPS for 24 h to establish an alveolar epithelial cell injury model and subsequently treated with CA or Ca2+/calmodulin-dependent protein kinase II (CaMKII) inhibitor KN93. Flow cytometry, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, and lactate dehydrogenase release assays were used to evaluate apoptosis, cell viability, and lactate dehydrogenase activity, respectively. Levels of inflammatory cytokines (interleukin-6, interleukin-1β, tumor necrosis tactor-α, and interferon-γ) and oxidative stress markers (reactive oxygen species, superoxide dismutase, catalase, and malondialdehyde) were determined using enzyme-linked immunosorbent assay and specific assay kits, respectively. Furthermore, levels of apoptosis-related proteins (cleaved caspase-3, Bcl-2-associated X, and Bcl-2) and CaMKII were assessed via western blotting. CA did not exhibit significant cytotoxicity in A549 cells. It dose-dependently improved the cell viability, suppressed apoptosis, decreased cleaved caspase-3 and Bcl-2-associated X levels, and increased Bcl-2 levels in LPS-treated A549 cells. It also inhibited inflammatory factor release and oxidative stress in LPS-induced A549 cells. Similar results were observed in the KN93- and CA-treated groups. Western blotting assay revealed that CA and KN93 inhibited CaMKII pathway activation, as indicated by the reduced p-CaMKII and p-phospholamban (PLN) levels and p-CaMKII/CaMKII and p-PLN/PLN ratios. Overall, CA alleviated alveolar epithelial cell injury by inhibiting the inflammatory response and oxidative stress and inducing cell apoptosis in LPS-induced A549 cells by regulating the CaMKII pathway, serving as a potential candidate for ALI prevention and treatment.

Sex-specific decline in prefrontal cortex mitochondrial bioenergetics in aging baboons correlates with walking speed.

Mitochondria play a crucial role in brain aging due to their involvement in bioenergetics, neuroinflammation and brain steroid synthesis. Mitochondrial dysfunction is linked to age-related neurodegenerative diseases, including Alzheimer's disease and Parkinson's disease. We investigated changes in the activities of the electron transport chain (ETC) complexes in normally aging baboon brains and determined how these changes relate to donor sex, morning cortisol levels, and walking speed. Using a novel approach, we assessed mitochondrial bioenergetics from frozen prefrontal cortex (PFC) tissues from a large cohort (60 individuals) of well-characterized aging baboons (6.6-22.8 years, approximately equivalent to 26.4-91.2 human years). Aging was associated with a decline in mitochondrial ETC complexes in the PFC, which was more pronounced when activities were normalized for citrate synthase activity, suggesting that the decline in respiration is predominantly driven by changes in the specific activity of individual complexes rather than changes in mitochondrial number. Moreover, when donor sex was used as a covariate, we found that mitochondrial respiration was preserved with age in females, whereas males showed significant loss of ETC activity with age. Males had higher activities of each individual ETC complex and greater lactate dehydrogenase activity relative to females. Circulating cortisol levels correlated only with complex II-linked respiration in males. We also observed a robust positive predictive relationship between walking speed and respiration linked to complexes I, III, and IV in males but not in females. This data reveals a previously unknown link between aging and bioenergetics across multiple tissues linking frailty and bioenergetic function. This study highlights a potential molecular mechanism for sexual dimorphism in brain resilience and suggests that in males changes in PFC bioenergetics contribute to reduced motor function with age.

Influence of Starvation on Biochemical, Physiological, Morphological, and Transcriptional Responses Associated with Glucose and Lipid Metabolism in the Liver of Javelin Goby (Synechogobius hasta).

In this study, the influence of fasting on hepatic glucose and lipid metabolism was explored by examining biochemical, antioxidative, and morphological indicators and transcriptional expression in the liver of javelin goby (Synechogobius hasta) after 0, 3, 7, or 14 days of starvation. Marked reductions in hepatic glycogen and triglycerides occurred from the seventh day of starvation until the end of the trial (p < 0.05). However, no alterations in hepatic cholesterol or protein were detected throughout the entire experiment (p > 0.05). During fasting, the activities of pyruvate kinase, lactate dehydrogenase, and glycogen phosphorylase a all rose firstly and then fell (p < 0.05). The activities of hepatic fatty acid synthase and acetyl-CoA carboxylase were minimized to their lowest levels at the end of food deprivation (p < 0.05), while lipase was elevated after 7-14 days of fasting (p < 0.05). Catalase, glutathione, and the total antioxidative capacity were increased and maintained their higher values in the later stage of fasting (p < 0.05), whereas malondialdehyde was not significantly changed (p > 0.05). Hepatic vein congestion, remarkable cytoplasmic vacuoles, and irregular cell shape were present in S. hasta which endured 3-7 days of fasting and were less pronounced when food shortage was prolonged. In terms of genes associated with glucose and lipid metabolism, the hepatic phosphofructokinase gene was constantly up-regulated during fasting (p < 0.05). However, the mRNA levels of glycogen synthase and glucose-6-phosphatase were obviously lower when the food scarcity extended to 7 days or more (p < 0.05). Fatty acid synthase, stearoyl-CoA desaturase 1, and peroxisome proliferator-activated receptor γ were substantially down-regulated in S. hasta livers after 7-14 days of food deprivation (p < 0.05). However, genes involved in lipolysis and fatty acid transport were transcriptionally enhanced to varying extents and peaked at the end of fasting (p < 0.05). Overall, starvation lasting 7 days or more could concurrently mobilize hepatic carbohydrates and fat as energy resources and diminished their hepatic accumulation by suppressing biosynthesis and enhancing catabolism and transport, ultimately metabolically and structurally perturbing the liver in S. hasta. This work presents preliminary data on the dynamic characteristics of hepatic glucose and lipid metabolism in S. hasta in response to starvation, which may shed light on the sophisticated mechanisms of energetic homeostasis in fish facing nutrient unavailability and may benefit the utilization/conservation of S. hasta.

Angiotensin 1–7 restrains vascular injury of extracorporeal membrane oxygenation by inhibiting ferroptosis

BackgroundAngiotensin 1–7 (Ang1-7) is the classical end product of angiotensin II, which has the effects of dilating blood vessels, protecting endothelial cells, anti-hypertension, improving cardiac function, and inhibiting atherosclerosis. We hypothesize that Ang1-7 inhibits human umbilical vein endothelial cells (HUVEC) ferroptosis through NF-κB/P53 signal pathway, and reduces extracorporeal membrane oxygenation (ECMO) vascular injury. MethodsCultured HUVEC were seeded into 15 wells and randomly divided into five groups: the control group and four experimental groups (erastin, erastin + Ang1-7, erastin + Ang1-7 + Betulinic acid, erastin + Betulinic acid). After stimulation, cell viability, lactate dehydrogenase (LDH), malondialdehyde (MDA), and superoxide dismutase (SOD) activity were measured. The effects of Ang1-7 on HUVEC microstructure, antioxidant enzymes (ferritin heavy chain 1 (FTH1), cystine/glutamic acid reverse transport solute carrier family 7 members 11 (SLC7A11 or XCT), superoxide dismutase-2 (SOD-2) and glutathione peroxidase 4 (GPX4)), NF-κB, P-NF-κB, P53, and P-P53). ResultsErastin stimulation promoted HUVEC lipid peroxidation, decreased antioxidant enzyme expression, increased P-NF-κB, P53, and P-P53 expressions, and damaged HUVEC mitochondrial structure. Ang1-7 alleviated the effect of erastin on HUVEC, which was destroyed by Betulinic acid. ConclusionAngiotensin1-7 pretreatment inhibited vascular endothelial cells’ ferroptosis and alleviated ECMO vessel injury through NF-κB /P53 signal pathway.

Understanding the mechanisms behind the antibacterial activity of magnesium hydroxide nanoparticles against sulfate-reducing bacteria in sediments

Nanomaterials, with their small size, surface characteristics, and antibacterial properties, are extensively employed across environmental, energy, biomedical, agricultural, and other industries. This study examined the antibacterial efficacy of magnesium hydroxide (Mg(OH)2) nanoparticles (NPs) against sulfate-reducing bacteria (SRB) within sediments. The inhibitory effects of two types of Mg(OH)2 NPs with distinct particle sizes (20.3 and 29.6 nm) and concentrations (0–10.0 mg/mL) were examined under optimal treatment conditions. The antibacterial mechanisms of Mg(OH)2 NPs through direct contact and dissolution effects were determined. The results revealed a correlation between the concentration, particle size, and inhibitory activity, with the smallest NPs (20.3 nm) at the highest concentration (10.0 mg/mL) substantially reducing SRB counts from 8.77 ± 0.18 to 6.48 ± 0.13 log10 colony forming units/mL after 6 h treatment. Treatment with high concentrations of Mg(OH)2 NPs induced cellular damage, reduced intracellular lactate dehydrogenase activity, and elevated intracellular catalase activity and H2O2 content, suggesting that the contact effect of NPs stimulated SRB. This leads to oxidative stress response and structural damage to the cell membrane, which has emerged as the primary driver of the antibacterial action of Mg(OH)2 NPs. This study presents a novel nanomaterial that can inhibit and control SRB in natural sedimentary environments.

NLRP3 targets HMGB1 to exacerbate the pyroptosis of canine corneal epithelial cells infected with Staphylococcus pseudintermedius

PurposeThis study focused on the mechanisms of pyroptosis and oxidative damage exacerbation by NOD-like receptor thermal protein domain associated protein 3 (NLRP3) during the infection of canine corneal epithelial cells (CCECs) with Staphylococcus pseudintermedius. MethodsThe CCECs treated with dimethyl fumarate (DMF), recombinant high mobility group protein 1 (HMGB1), or N-acetylcysteine (NAC). The gasdermin (GSDM) family and HMGB1 mRNA expression levels were detected using quantitative reverse transcription polymerase chain reaction. Lactate dehydrogenase activity, bacterial counts, the pyroptosis rate, reactive oxygen species (ROS) content, and antioxidant enzyme activity were used to reflect pyroptosis and oxidation level. ResultsRegulation of NLRP3 significantly affected the pyroptosis rate and GSDMD-N expression levels during S. pseudintermedius infection. Inhibition of GSDMD-N protein activation by DMF reversed the exacerbation of pyroptosis induced by NLRP3 overexpression and reduced the levels of cleaved interleukin-1β (IL-1β), cleaved cysteinyl aspartate-specific protease-1, and NLRP3. In addition, NLRP3 was found to target the HMGB1 promoter and regulate its protein expression, to increase ROS accumulation and GSDMD-N expression levels, and activate the NLRP3-HMGB1-ROS-GSDMD signaling axis to aggravate pyroptosis during infection. ConclusionsNLRP3 aggravates pyroptosis and oxidative damage associated with the activation of NLRP3-GSDMD and NLRP3-HMGB1-ROS-GSDMD signaling pathways during the infection of CCECs with S. pseudintermedius.

Protective Role of Ovothiol-A against Muscle and Kidney Injuries in Obese Rats

Background: Obesity is a serious chronic metabolic disease impairing health damaging many organs such as kidneys and muscles. Ovothiol-A (Ovo-A) has been found to keep the redox balance normal in sea urchins indicating its antioxidant characteristics. Aim: This study aims to investigate the protective effects of Ovo-A on kidneys and muscles in obese rats. objective: Investigation of the protective effect of Ovo-A on kidneys and muscles in obese rats. Methods: In-silco studies were performed on lactate dehydrogenase (LDH) and creatine kinase (CK) with Ovo-A to compute their binding affinities. Obesity was induced by high-fat diet (HFD) for 4 weeks. Wistar rats were used in this study as 6 rats per group as control, HFD, Ovo-A (200 and 400 mg/Kg, p.o) groups. Results: Docking results have revealed that Ovo-A has affinities to bind to LDH (-8.5 kcal/mol) and CK (-17.7 kcal/mol). Ovo-A reduced the levels of uric acid, urea, creatinine, LDH, CK, malondialdehyde (MDA), and nitric oxide (NO), while increasing the levels of glutathione (GSH), catalase (CAT), and glutathione-S-transferase (GST). Histopathological investigations have revealed that Ovo-A restored the renal and muscular structure. Conclusion: The current study showed that Ovo-A has a protective effect on kidneys and muscles in obese rats. Ovo-A enhances renal and muscular functions by inhibiting LDH and CK activities and improving the antioxidant system. Ovo-A is more effective in the high dose.

Regulatory mechanism of antioxidant enzymes on microbial metabolism and NADH in anaerobic fermentation of food waste for hydrogen production

Anaerobic fermentation is frequently hampered by toxicity arising from oxidative stress, and antioxidant enzymes play a crucial role in combating oxidative stress. In this study, the mechanism of microbial consortium and metabolic pathways regulated by antioxidant enzyme genes in anaerobic fermentation with different pH values was revealed. The results showed that antioxidant enzyme genes, such as glutathione peroxidase, peroxidase, and superoxide dismutase, were 5 times, 3 times, and 2 times higher at pH 7 than at pH 5, respectively. This allowed Clostridium to effectively resist oxidative stress, with substrate metabolism dominated by glycolysis, leading to an increase in the NADH/NAD+ ratio. Furthermore, the relative abundance of hydrogenase and electron transfer efficiency increased by 4.5 times and 2.71 times, respectively, resulting in a hydrogen production of 21.48 L/L. When antioxidant enzyme genes were inhibited at pH 5, the substrate mainly produced biomolecules for combating oxidative stress through the pentose phosphate and glycerophospholipid pathways, and led to the transformation of dominant genus into Lactobacillus. With an increased lactate dehydrogenase activity of 5.34 times, the final lactate production reached 65.17 g/L, which was 19.69 times higher than at pH 7. These results elucidate the regulatory mechanism of pH mediated antioxidant enzyme genes on hydrogen production and improve the controllability of anaerobic fermentation.

The Effect of Marginal Zn2+ Excess Released from Titanium Coating on Differentiation of Human Osteoblastic Cells.

Composite coatings based on chitosan and zinc nanoparticles (ZnNPs) were successfully produced on Ti13Zr13Nb substrates by cathodic electrophoretic deposition (EPD). The unfavorable phenomenon of water electrolysis-induced nonuniformity was reduced by applying a low voltage (20 V) and a short deposition time (1 min). Surface analysis (roughness and hydrophilicity) reveals the potential of these coatings for enhancing cell attachment and bone-implant integration. However, there is a concern about adhesion and strength; therefore, incorporating ZnNPs shows promise for enhancing mechanical properties, suggesting opportunities for further optimization of the process. The aim of this work was to investigate whether Zn2+ released from coating yields overt cellular impairment. hFOB1.19 osteoblastic cells were used as a model in this study. A subtoxic, 0.125 mmol/L, Zn concentration did not cause significant negative changes in cultured osteoblastic cells, as there was no significant change in their viability and their mitochondrial metabolism. Moreover, the alkaline phosphatase and lactate dehydrogenase activities were aggravated. However, a high, over 0.175 mmol/L, Zn2+ concentration caused total cell death. This was caused by the inhibition of mitochondrial enzymes' activities. Our data indicate that composite coatings releasing Zn2+ may be used as the differentiating factor toward osteoblastic cells.

Minocycline alleviates lipopolysaccharide-induced cardiotoxicity by suppressing the NLRP3/Caspase-1 signaling pathway.

Minocycline (Min), as an antibiotic, possesses various beneficial properties such as anti-inflammatory, antioxidant, and anti-apoptotic effects. Despite these known qualities, the precise cardioprotective effect and mechanism of Min in protecting against sepsis-induced cardiotoxicity (SIC) remain unspecified. To address this, our study sought to assess the protective effects of Min on the heart. Lipopolysaccharide (LPS) was utilized to establish a cardiotoxicity model both in vivo and in vitro. Min was pretreated in the models. In the in vivo setting, evaluation of heart tissue histopathological injury was performed using hematoxylin and eosin (H&E) staining and TUNEL. Immunohistochemistry (IHC) was employed to evaluate the expression levels of NLRP3 and Caspase-1 in the heart tissue of mice. During in vitro experiments, the viability of H9c2 cells was gauged utilizing the CCK8 assay kit. Intracellular ROS levels in H9c2 cells were quantified using a ROS assay kit. Both in vitro and in vivo settings were subjected to measurement of oxidative stress indexes, encompassing glutathione (GSH), malondialdehyde (MDA), and superoxide dismutase (SOD) levels. Additionglly, myocardial injury markers like lactate dehydrogenase (LDH) and creatine kinase MB (CK-MB) activity were quantified using appropriate assay kits. Western blotting (WB) analysis was conducted to detect the expression levels of NOD-like receptor protein-3 (NLRP3), caspase-1, IL-18, and IL-1β, alongside apoptosis-related proteins such as Bcl-2 and Bax, and antioxidant proteins including superoxide dismutase-1 (SOD-1) and antioxidant proteins including superoxide dismutase-1 (SOD-2), both in H9c2 cells and mouse heart tissues. In vivo, Min was effective in reducing LPS-induced inflammation in cardiac tissue, preventing cell damage and apoptosis in cardiomyocytes. The levels of LDH and CK-MB were significantly reduced with Min treatment. In vitro studies showed that Min improved the viability of H9C2 cells, reduced apoptosis, and decreased ROS levels in these cells. Further analysis indicated that Min decreased the protein levels of NLRP3, Caspase-1, IL-18, and IL-1β, while increasing the levels of SOD-1 and SOD-2 both in vivo and in vitro. Min alleviates LPS-induced SIC by suppressing the NLRP3/Caspase-1 signalling pathway in vivo and in vitro.

F13B regulates angiogenesis and tumor progression in hepatocellular carcinoma via the HIF-1α/VEGF pathway.

Hepatocellular carcinoma (HCC) is a highly aggressive malignant tumor with a poor prognosis. This research aimed to investigate the role of F13B in HCC and its underlying mechanisms. Through comprehensive bioinformatics analysis of the GSE120123 and The Cancer Genome Atlas (TCGA)-Liver hepatocellular carcinoma (LIHC) datasets, we identified 220 overlapping prognosis-related genes. Eight key genes, including the previously unreported CCDC170 and F13B in HCC, were identified through Least Absolute Shrinkage and Selection Operator (LASSO)-Cox regression analysis. F13B emerged as a significant prognostic factor in HCC, warranting further investigation in subsequent analyses. In vitro experiments showed that F13B expression was notably reduced in HCC cell lines and tissues, particularly in Huh-7 and SMMC-7721 cells. Overexpression of F13B inhibited cell invasion, migration, and proliferation, while its knockdown produced the opposite effect. A lactate dehydrogenase (LDH) activity assay in human umbilical vein endothelial cells (HUVECs) demonstrated that F13B overexpression reduced vascular endothelial growth factor (VEGF)-induced cytotoxicity, whereas knockdown increased it. Further analysis revealed that F13B negatively regulates VEGFA expression, affecting HUVEC proliferation. In HUVECs, F13B overexpression reversed VEGF-induced upregulation of key angiogenesis markers, including phospho-VEGF receptor 2 (p-VEGFR2), matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 (MMP-9), as well as AKT/mTOR signaling proteins, phospho-Akt (p-AKT), and phospho-mTOR (p-mTOR). Additionally, F13B negatively regulated VEGFA and hypoxia-inducible factor 1 A (HIF1A) under hypoxic conditions, counteracting the hypoxia-induced increase in cell viability. These findings suggest that F13B regulates angiogenesis through the HIF-1α/VEGF pathway and plays a crucial role in HCC progression. Our results highlight the potential of F13B as a therapeutic target in HCC, providing novel insights into the molecular mechanisms of HCC and its prognostic significance.