MDA Content Research Articles

Exogenous melatonin and grafting improved the growth and physiological characteristics of Citrullus lanatus seedlings under cadmium stress.

Citrullus lanatus is an important vegetable crop, but it is heavily polluted by cadmium. In this study, we used C. Lanatus as experimental material to investigate effects of different concentrations (0, 50, 100, 200, 400µmol.L-1) of exogenous melatonin, and grafting on the physiological growth index and anatomical structure of seedlings were studied by simulating Cd2+ (180mg L-1) stress environment. The results showed that exogenous melatonin could reduce the injury degree of C. Lanatus seedlings under cadmium stress, and the best effect was obtained when the concentration is 100µmol.L-1. Compared with the control treatment (0µmol.L-1 melatonin), the number of leaves treated with 100µmol.L-1 melatonin increased by 32.61%, the density of mesophyll tissue decreased by 16.47%, the relative content of chlorophyll in seedling leaves increased by 42.90%, and the activity of superoxide dismutase (SOD) and the content of soluble protein increased by 28.64% and 60.11%, respectively. In addition, grafting could effectively alleviate the damage of cadmium stress to C. lanatus seedlings, and the effect of 100µmol.L-1 melatonin and grafting interaction on seedlings was higher than that of single melatonin or grafting treatment. Under cadmium stress, compared with the self-rooted seedlings of melatonin (0µmol.L-1), the stem diameter and the diameter of main vein increased by 55.56% and 34.12%, and the water content of leaf tissue increased by 93.55%. In addition, the relative conductivity and MDA content decreased by 54.45% and 53.07%, respectively. Therefore, there was a significant interaction between exogenous melatonin and grafting in alleviating the injury of C. lanatus seedlings under cadmium stress. Our research explored the optimal concentration of exogenous melatonin to alleviate the injury of C. lanatus seedlings under cadmium stress that provided theoretical basis for revealing the mechanism of exogenous melatonin to improve the resistance of C. lanatus to cadmium stress.

Effect of Gardeniae Fructus Powder on Growth Performance, Antioxidant Capacity, Intestinal Barrier Function, and Colonic Microbiota of Weaned Piglets

The present study aimed to explore the effect of GF powder on the growth performance, diarrhea rate, antioxidant and immune capacity, and intestinal health of weaned piglets. A total of 144 weaned piglets (8.29 ± 0.11 kg) at 21 d old were randomly assigned to four groups, with each treatment consisting of six replicate pens, with six piglets per pen, and each pen containing three barrows and three gilts. The piglets were fed a basal diet supplement with 0%, 0.4%, 0.6%, and 0.8% GF powder (n = 36). Our results indicated that compared with the basal diet, the F/G and diarrhea rate were remarkably decreased in the 0.8% GF group (p < 0.05). Serum biochemical parameters showed that supplementation with GF significantly increased the content of HDL-C (0.6 and 0.8% levels), IL-6 (0.8% level), IL-10 (0.4, 0.6, and 0.8% levels), Ig G (0.4% level), and Ig A (0.8% level) compared with the basal diet (p < 0.05). The index of antioxidant capacity showed that compared with a basal diet, supplementation with GF significantly decreased serum MDA content (0.4% and 0.8% levels) and jejunal and ileal MDA content (0.4%, 0.6%, and 0.8% levels) (p < 0.05). Additionally, compared with the basal diet, supplementation with GF significantly increased serum and ileal T-AOC content (0.4%, 0.6%, and 0.8% levels), serum T-SOD content (0.4% and 0.8% levels), ileal T-SOD content (0.4%, 0.6%, and 0.8% levels), CAT content (0.4%, 0.6%, and 0.8% levels), and jejunal GSH-Px content (0.8% level) (p < 0.05). The results of gene expression indicate that compared with the basal diet, supplementation with GF significantly increased Nrf 2 (0.4% level), NQO (0.4% level), SOD 1 (0.4% and 0.8% levels), and GCLC (0.4% level) and GCLM (0.8% level) abundance in jejunal mucosa; supplementation with GF significantly increased Nrf 2 (0.4%, 0.6%, and 0.8% levels), HO-1 (0.4% level), NQO (0.8% level), SOD 1 (0.4% and 0.8% levels), and GCLC (0.4% level) and GCLM (0.8% level) abundance in ileal mucosa (p < 0.05). Ulteriorly, the present results indicate that supplementation with GF at the 0.8% level significantly increased the villus height in the jejunum and ileum as well as the villus/crypt ratio in the ileum compared with the basal diet (p < 0.05). Compared with the basal diet, 0.4% GF significantly increased Occludin gene expression in ileal mucosa (p < 0.05), 0.6% GF significantly increased ZO-1, Claudin-1, and Occludin gene expression in jejunal mucosa (p < 0.05), and 0.8% GF significantly increased ZO-1 and Occludin gene expression in jejunal mucosa along with Occludin expression in ileal mucosa (p < 0.05). Furthermore, colonic microbiota composition showed that Shannon, observed species, and Chao 1 indices were significantly increased in the 0.8% GF group compared with the basal diet (p < 0.05). At the phylum level, in comparison with the basal diet, the relative abundance of Firmicutes significantly decreased in the 0.4%, 0.6%, and 0.8% GF groups, and Bacteroidetes increased in the 0.8% GF group (p < 0.05). At the genus level, compared with the basal diet, 0.6% and 0.8% GF significantly increased Prevotella abundance, and 0.6% GF significantly decreased Coprococcus abundance (p < 0.05). At the species level, compared with the basal diet, 0.8% GF significantly increased Prevotella copri abundance, and 0.4%, 0.6%, and 0.8% GF significantly decreased Blautia obeum abundance (p < 0.05). In summary, a dietary supplement with 0.8% Gardeniae Fructus powder significantly decreased the F/G and diarrhea rate and improved antioxidant capacity and intestinal barrier function, which may be associated with the improvement of the relative abundance of Prevotella copri. These findings indicate that Gardeniae Fructus powder may be used as a feed additive in swine weaning.

Formulation and Characterization of Sodium Caseinate/Phloretin Complexes as Antioxidant Stabilizers in Oil-in-Water Emulsions

Emulsifiers with antioxidant properties, such as protein/polyphenol complexes, adsorb at the oil-water interface and improve the physical and oxidative stability of emulsions. Here, 2% (w/w) sodium caseinate and varying concentrations of phloretin (0–10 mM) were used to stabilize oil-in-water emulsions. Control emulsions with protein alone showed poor stability with increased droplet sizes from 0.33 µm to 5.18 µm after 30 days, while no significant change was observed in emulsions containing phloretin (remaining below 400 nm). The in vitro antioxidant activities increased with increasing phloretin concentrations (0 to 10 mM). In the ABTS assay, the antioxidant activity improved from 14.02 ± 8.33% to 95.09 ± 1.31%, and in the DPPH assay, it increased from 32.59 ± 2.73% to 99.03 ± 0.14%. Similarly, the oxidative stability of the emulsions improved with increasing phloretin concentrations (0 to 10 mM). After 30 days of storage, PV decreased from 38.22 ± 2.58 µM to 11.81 ± 2.55 µM, and MDA content reduced from 48.43 ± 0.31 µM to 7.24 ± 0.21 µM. Measuring the apparent viscosity demonstrated a reduction in viscosity with the addition of phloretin. These findings demonstrate that incorporating phloretin into sodium caseinate-stabilized emulsions as a novel antioxidant emulsifier can be an effective strategy to extend the shelf life of emulsified food products prone to oxidative deterioration.

Effects of Dietary Nano-Composite of Copper and Carbon on Antioxidant Capacity, Immunity, and Cecal Microbiota of Weaned Ira White Rabbits

This experiment investigated the effects of dietary supplementation with nano-composites of copper and carbon (NCCC) on antioxidants, immune functions, and the cecum microbiota of weaned Ira white rabbits. A total of 240 weaned 35-day-old Ira white rabbits were randomly allocated to five dietary treatments (n = 6 per treatment, each replicate consisted of eight rabbits) that included the control group (CON) with a basal diet, the SAL group with 60 mg/kg salinomycin (SAL) in addition to the basal diet, and the NCCC I, II, III groups, which were supplemented with 50, 100, and 200 mg/kg NCCC, respectively, in addition to the basal diet. The test lasted for 28 d. The results showed that dietary NCCC supplementation increased the liver Cu/Zn-SOD content and up-regulated the gene expression of Cu/Zn-SOD (p < 0.05), while also reducing the content of MDA in the liver and enhancing the antioxidant capacity of Ira white rabbits. Moreover, the NCCC diet supplementation reduced the content of IL-6 and down-regulated the relative expression of IL-6 and IL-1β genes in the jejunum of Ira white rabbits (p < 0.05). In addition, the metagenomic analysis of 16 S rRNA showed significant differences in the cecal microbial structure of weaned Ira white rabbits in the NCCC III group compared with the CON, NCCC I, and NCCC II groups (p < 0.05). Firmicutes and Bacteroidetes were the dominant phyla of cecal microorganisms in weaned Ira rabbits in the NCCC diet groups. The dominant genera included unidentified Eubacteriaceae, unclassified Lachnospiraceae, Christensenellaceae, and Ruminococcus. Furthermore, the relative abundance of Ruminococcus in the NCCC I and II groups was lower than that in the CON group in the cecum of Ira white rabbits (p < 0.05). In summary, our results showed that diet supplementation with NCCC could enhance the antioxidant capacity in the liver, alleviate intestinal inflammation, and regulate the structure of intestinal flora, improving the health of Ira white rabbits.

PcNAC25, a NAC transcription factor of Pugionium cornutum(L.) Gaertn conferring enhanced drought and salt stress tolerances in Arabidopsis

Pugionium cornutum (L.) Gaertn (P. cornutum) has strong tolerance to drought, salt and disease, but the tolerance mechanisms for such stresses in P. cornutum are largely unknown. In this study, we identified the PcNAC25 transcription factor gene in P. cornutum. Its open reading frame was revealed to comprise 891 bp, encoding a protein consisting of 297 amino acids, with an isoelectric point of 6.61. Phylogenetic analysis showed that PcNAC25 was most closely related to ANAC019. The expression of PcNAC25 was induced by dehydration, mannitol, heat, cold, salt stresses and abscisic acid (ABA), salicylic acid (SA), and methyl jasmonate (JA) treatments. A subcellular localization analysis confirmed that PcNAC25 was localized in the nucleus. The overexpressing PcNAC25 lines in Arabidopsis had longer roots than wild-type (WT) lines under drought and salt stress. The overexpression of PcNAC25 improved drought and salt tolerance in transgenic Arabidopsis. Under drought and salt stress, PcNAC25 transgenic lines exhibited higher the CAT, POD and SOD activities and scavenging ability of hydroxyl radical than WT, more proline accumulation than WT and less MDA and H2O2 content and superoxide anion production rate than WT. PcNAC25 transgenic lines also exhibited greater reduced water loss rate of detached leaves than WT. Meanwhile, DAB and NBT staining showed that the accumulation of hydrogen peroxide and superoxide anion in PcNAC25 transgenic lines were also less than WT. In addition, overexpressing PcNAC25 enhanced the expression of drought response genes (DREB2A, SOD4, RD29A, NCED3, POD3, P5CS1, PYR1 and SAG13) and salt response genes NHX, SLAH1, SOS1 and NPF6.3. The mentioned above results indicated that PcNAC25 is a positive regulator that activates ROS-scavenging enzymes and enhances root formation in Arabidopsis, which provided a basis for further research on the molecular mechanism of PCNAC25-mediated regulation of drought and salt stress, and also provided gene resources of drought and salt tolerance.

Potential Therapeutic Effect of Vitamin C on Methotrexate- Induced Damage in the Cerebral Cortex.

<b>Background and Objective:</b> Methotrexate is an anti-metabolic medication used to treat cancer. It causes oxidative stress in nerve tissue and has neurotoxic effects. A strong antioxidant and effective free radical scavenger is vitamin C. The current research aims to investigate the potential protective impact of vitamin C and the toxic consequences of methotrexate. <strong>Materials and Methods:</strong> Thirty-six rats were used in this research and group one (Group 1) got no treatment at all. For 4 weeks, (Group 2) underwent a single intraperitoneal injection of methotrexate at a dose of 20 mg/kg once a week and (Group 3) got methotrexate at the same dosage as Group 2 and vitamin C (20 mg/kg) intragastrically every other day for four weeks. Rats were killed after the experiment and brain hemispheres were removed and prepared for light microscopic analysis. The cerebral hemispheres were ready for biochemical analysis to determine the brain tissue's concentrations of MDA, CAT, GSH and SOD. Data analysis was conducted using SPSS software version 20. <b>Results:</b> In the methotrexate (2)-treated group, there were histological alterations manifested as a reduction in granular layer thickness. Purkinje cells exhibit a reduction in number, a shrinking of the cell bodies and a loss of monolaminar organization. Reduced cellularity was seen in the molecular layer. These cellular alterations are lessened and the thickness of the granular and molecular cell layers is restored following vitamin C treatment. When compared to the MTX+Vitamin C group, vitamin C greatly attenuates the biochemical and histological alterations caused by MTX. <b>Conclusion:</b> Results concluded that although methotrexate is a toxic medication that damages the brain cortex, its toxicity is reduced when vitamin C is taken with it.

Bacillus coagulans alleviates hepatic injury caused by Klebsiella pneumoniae in rabbits.

As an opportunistic bacterial pathogen, Klebsiella pneumoniae (KP) is prone to causing a spectrum of diseases in rabbits when their immune system is compromised, which poses a threat to rabbit breeding industry. Bacillus coagulans (BC), recognized as an effective probiotic, confers a variety of benefits including anti-inflammatory and antioxidant properties. The purpose of this study was to investigate whether dietary BC can effectively alleviate hepatic injury caused by KP. In this study, the rabbits were initially pretreated with varying doses of BC (1×106, 5×106, and 1×107 CFU/g), followed by a challenge with KP at a concentration of 1011 CFU/mL. Liver tissues were harvested and processed for histological assessment using H&E and VG stains to assess structural alterations. Biochemical assays were employed to quantify the enzymatic activities of T-SOD and GSH-Px, as well as the MDA content. Furthermore, ELISA was utilized to detect the levels of inflammatory cytokine (IL-10, IL-6, IL-1β and TNF-α) and apoptotic-related gene (Bcl-2, Bax). Morphological observation indicated that BC can effectively mitigate KP-induced hepatic sinusoidal dilatation and congestion, as well as ameliorate the degree of hepatic fibrosis. Further analysis showed that BC significantly lowered MDA level in KP-treated rabbits, while enhanced the activities of T-SOD and GSH-Px. Additionally, ELISA result showed that BC pretreatment significantly reduced the levels of pro-inflammatory cytokines TNF-a, IL-6, IL-1β and pro-apoptotic gene Bax, while increasing the levels of anti-inflammatory cytokine IL-10 and anti-apoptotic gene Bcl-2 in KP-treated rabbits. Above data indicate that BC supplementation effectively attenuated oxidative stress and inflammatory response induced by KP through augmenting the activities of antioxidant enzymes and diminishing the levels of pro-inflammatory factors. Furthermore, it reduced the Bax/Bcl-2 ratio in the liver, thereby inhibiting KP-induced apoptosis. The treatment group receiving 5x106 CFU/g BC benefitted most from the protective effect.

FOXA2 regulates endoplasmic reticulum stress, oxidative stress, and apoptosis in spermatogonial cells by the Nrf2 pathway under hypoxic conditions.

Hypoxia-caused spermatogenesis impairment may contribute to male infertility. FOXA2 has been found to be abundant in spermatogonial stem cells and critical for spermatogenesis. Here we aimed to explore the roles of FOXA2 in regulating spermatogonial cells against hypoxia stimulation. Our results showed that FOXA2 expression was downregulated in hypoxia-stimulated spermatogonial cells. Overexpression of FOXA2 prevented hypoxia-induced endoplasmic reticulum (ER) stress with decreased expression levels of associated markers including GRP78, CHOP, and ATF-4. FOXA2 overexpression caused a decrease in MDA content and an increase in activities of SOD, CAT, and GSH-Px in spermatogonial cells under hypoxic conditions, implying its inhibitory effect on oxidative stress. Besides, cell apoptosis under hypoxic conditions was also prevented by FOXA2 overexpression, as shown by reduced apoptotic rate and caspase-3 activity. Moreover, we found that hypoxia stimulation inactivated the Nrf2 pathway, which could be prevented by FOXA2 overexpression. Nrf2 knockdown attenuated the effects of FOXA2 overexpression on hypoxia-induced ER stress, oxidative stress, and apoptosis in spermatogonial cells. In conclusion, FOXA2 exerted protective effects on spermatogonial cells against hypoxia-induced ER stress, oxidative stress, and apoptosis via regulating Nrf2/HO-1 signaling. These findings suggested that FOXA2 might be a therapeutic target for treating hypoxia-induced spermatogenesis impairment.

Hepatoprotective efficacy of Argemone mexicana L. root in paracetamol-induced hepatotoxicity in a rat model.

Argemone mexicana L. (Papaveraceae), a weed that thrives in the tropical and subtropical areas of South and Central America, Mexico, Caribbean Islands and India. In India, it has been used traditionally to treat vesicular calculus, inflammatory conditions, and hepatobiliary disorders. The present study was aimed to investigate the hepatoprotective efficacy of A. Mexicana roots in paracetamol (PCM)-induced toxicity rat. The methanol extract of A. mexicana (MEAM) root was analyzed using Gas Chromatography-Mass Spectrometry (GC-MS) analysis to identify its compounds. Molecular docking analysis of the compounds was conducted against TGF-β and PPAR-α. The hepatoprotective activity of MEAM (200, 400mg/kg) was evaluated in PCM (3000mg/kg) intoxicated rats by measuring serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), lactate dehydrogenase (LDH), total bilirubin (TB), total protein (TP), albumin (ALB), globulin (GLB), total cholesterol (TC), triglyceride (TG), low-density lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C). Silymarin (100mg/kg) was used as reference drug. Oxidative stress biomarkers such as superoxide dismutase (SOD), catalase (CAT), reduced glutathione (GSH), and lipid peroxidation (LPO) were investigated using liver homogenate. Additionally, the levels of pro-inflammatory cytokines including tumor necrosis factor-alpha (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), and anti-inflammatory cytokines interleukin-4 (IL-4) and interleukin-10 (IL-10) were studied. The results of the study were supported by histopathological examination. GC-MS analysis revealed 163 compounds, from which eleven compounds were selected based on their docking scores against TGF-β and PPAR-α. MEAM (400mg/kg) demonstrated a remarkable reduction in ALT, AST, ALP, GGT, and LDH in contrast to the PCM intoxicated group. A remarkable decline in TB and GLB, along with an increase in TP and ALB, was observed in the MEAM (400mg/kg) group compared to the untreated PCM group. Rats receiving MEAM (400mg/kg) exhibited a noticeable decrease in TC, TG, and LDL-C, along with an increase in HDL-C levels compared to PCM-induced untreated rats. The higher dose of MEAM also resulted in a significant decrease in TNF-α, IL-1β, and IL-6, and an increase in IL-4 and IL-10. Similarly, a notable elevation in SOD, CAT, and GSH, along with a decrease in MDA content, was observed in the group receiving MEAM (400mg/kg). The histopathological result showed reduction of sinusoidal space and vesicular nuclei, with improvement of hepatocytes at the dose of MEAM (400mg/kg). In molecular docking study, Eupatilin exhibited the highest docking scores of -10.4kcal/mol and -9.1kcal/mol against TGF-β and PPAR-α, respectively. MEAM root at dose of 400mg/kg exhibited hepatoprotective effect against PCM-induced toxicity rat. Eupatilin might be considered as a potential candidate for the hepatoprotective effect of A. mexicana root.

Dietary Se-enrich Cardamine violifolia supplementation decreases lipid deposition and improves antioxidant status in the liver of aging laying hens

Aging-related lipid metabolic disorder is related to oxidative stress. Selenium (Se)-enriched Cardamine violifolia (SEC) is known for its excellent antioxidant function. The objective of this study was to evaluate the effects of SEC on antioxidant capacity and lipid metabolism in the liver of aged laying hens. A total of 450 sixty-five-wk-old Roman laying hens were randomly divided into 5 treatments: a basal diet (without Se supplementation, CON) and basal diets supplemented with 0.3 mg/kg Se from sodium selenite (SS), 0.3 mg/kg Se from Se-enriched yeast (SEY), 0.3 mg/kg Se from SEC (SEC), or 0.3 mg/kg Se from SEC and 0.3 mg/kg Se from SEY (SEC + SEY). The experiment lasted for 8 wk. The results showed that dietary SEC + SEY supplementation decreased (P < 0.05) triglyceride (in the plasma and liver) and total cholesterol levels (in the plasma), and increased (P < 0.05) HDL-C concentration in plasma compared to CON diet. Compared with CON diet, SEC and/or SEY supplementation decreased (P < 0.05) the mRNA expression of hepatic ACC, FAS and HMGCR, and increased (P < 0.05) PPARα, VTG-II, Apo-VLDL II and ApoB expression. Dietary SEC + SEY and SEY supplementation increased (P < 0.05) Se content in egg yolk and breast muscle compared to CON diet. Dietary SEC, SEY or SEC + SEY supplementation increased (P < 0.05) the activity of antioxidant enzymes (GSH-PX, T-AOC and T-SOD) in the plasma and liver and decreased (P < 0.05) MDA content in the plasma compared to CON diet. Dietary Se supplementation promoted (P < 0.05) mRNA expression of Nrf2 in the liver. In contrast, dietary SEY and SEC supplementation resulted in a decrease (P < 0.05) of hepatic Keap1 mRNA expression compared to CON diet. Dietary SEC + SEY and/or SEC supplementation increased (P < 0.05) mRNA expression of Selenof, GPX1 and GPX4 in the liver compared with CON diet. In conclusion, dietary SEC (0.3 mg/kg Se) or SEC (0.3 mg/kg Se) + SEY (0.3 mg/kg Se) improved the antioxidant capacity and the lipid metabolism in the liver of aged laying hens, which might be associated with regulating Nrf2/Keap1 signaling pathway.

Gradient experiment reveals physiological stress from heavy metal zinc on the economically valuable seaweed Sargassum fusiforme.

Zn is a common heavy metal pollutant in water bodies and accounts for the largest proportion of heavy metal pollutants in many rivers entering the sea. This study investigated the growth and physiological response characteristics of Sargassum fusiforme under different divalent Zn ion concentration gradients. We observed that low concentration Zn2+ treatment (<2mgL-1) exerted no significant effect on the growth rate, photosynthesis, and nitrogen metabolism-related indicators of S. fusiforme. Treatment with medium to high Zn2+ concentrations (2-25mgL-1) significantly affected the growth rate, photosynthetic activity, nitrogen absorption rate, antioxidant enzyme activity, membrane lipids, and DNA peroxidation damage-related indicators of S. fusiforme. Under medium-to-high concentration treatments, the SOD activity of S. fusiforme decreased with increasing concentration, and the CAT activity increased with increasing treatment concentration. The MDA and H2O2 contents increased with increasing Zn2+ concentrations. At a Zn2+ concentration of 5mgL-1, the relative conductivity of S. fusiforme significantly increased. Treatment with higher Zn2+ concentrations significantly increased the 8-hydroxydeoxyguanosine (8-OHdG) content, poly ADP-ribose polymerase (PARP) activity, and Histone H2AX content of S. fusiforme, thus indicating that Zn2+ stress causes DNA damage. All Zn2+ concentrations induced mannitol accumulation, and soluble protein content decreased with increasing Zn2+ concentration. In summary, we observed that a Zn2+ concentration of 2-5mgL-1 may be the critical value for the response of S. fusiforme to Zn2+ stress. Higher concentrations of Zn in the environment can exert toxic effects on the growth, development, and biomass accumulation of S. fusiforme. This study provides a reference for the risk assessment and aquaculture management of seaweeds.

CircHOMER1 Alleviates Sevoflurane-Induced Hippocampal Neuronal Injury via Targeted Negative Regulation of miR-217.

Sevoflurane (Sev) is a widely applied anesthetic in clinical practice; however, it could induce neurotoxicity and lead to postoperative cognitive dysfunction (POCD). This study aimed to investigate the role and underlying mechanism of circHOMER1 in Sev-induced neurotoxicity and POCD. Sev treated mouse hippocampal neuronal HT22 cells and SD rats. RT-qPCR was used to detect the levels of circHOMER1 and miR-217. ELISA was employed to measure the levels of inflammatory factors IL-6, IL-1β, and TNF-α. Commercially available kits assessed the concentration of MDA and measured the activities LDH and SOD. The CCK-8 assay assessed cell viability. Flow cytometry analyzed cell apoptosis. The Morris water maze test evaluated the learning and cognitive abilities of the rats. Dual luciferase reporter assays and RIP experiments validated the targeted binding of circHOMER1 to miR-217. Sev treatment significantly reduces cell viability, increases apoptosis, stimulates inflammatory responses and oxidative stress, and induces learning and memory impairments in SD rats. Following exposure to Sev, the expression of circHOMER1 is markedly decreased, while overexpression of circHOMER1 can alleviate Sev-induced neuroinflammation, oxidative stress, and learning and memory deficits in rats. CircHOMER1 targets miR-217, and transfection of miR-217 antagonizes the neuroprotective effects of circHOMER1. This study demonstrated that circHOMER1 negatively regulated miR-217, thereby inhibiting Sev-induced neurotoxicity and learning and memory disorders.

2-Ethylhexyl diphenyl phosphate induces lung oxidative stress and pyroptosis in chicks.

2-Ethylhexyl diphenyl phosphate (EHDPHP) is a widely used organophosphorus flame retardant and plasticizer easily released into the environment. Its biological toxicity is of great concern. The lung is considered a possible target organ for EHDPHP, but currently, there are limited studies on the biotoxicity of EHDPHP in poultry lungs. Therefore, the lungs were selected as the target organ to study the toxic effects of EHDPHP on chicks and their mechanisms of action. In this study, 7-day-old chicks were gavaged with different concentrations of EHDPHP, and lung samples were collected at 14, 28, and 42days after intragastric administration. Lung histopathological and ultrapathological changes were examined by paraffin section-HE staining and transmission electron microscopy. The levels of lung damage markers (LDH) and oxidative stress markers (GSH-Px, SOD, and MDA) were detected by applying the kit. In contrast, lung cell pyroptosis-related factors (NLRP3, ASC, NF-κB, Pro-Caspase-1, IL-18, and IL-1β) and inflammatory factors (IL-6 and TNF-α) were assessed by using the qRT-PCR, Western blot and ELISA techniques. The results showed that EHDPHP induced pathological morphological changes and elevated LDH content in chick lungs, decreased lung antioxidant enzymes (GSH-Px and SOD) activities, increased peroxidation product MDA content and up-regulated the expression levels of cellular pyroptosis factors (NLRP3, ASC, NF-κB, Pro-Caspase-1, IL-18, and IL-1β), and the synthesis and secretion of inflammatory factors (IL-6 and TNF-α) were promoted. The above changes were EHDPHP dose-dependent. The results indicated that EHDPHP induced oxidative stress in chick lungs, resulting in oxidative damage to the lungs, and, intriguingly, the cellular pyroptosis pathway was activated, which was also involved in the process of EHDPHP-induced inflammatory damage in chick lungs. The results of this study revealed for the first time the damaging effects and mechanisms of EHDPHP on chick lungs. Also, they provided a scientific basis for further exploring the mechanisms of toxicity damage, safe use, and pollution control of EHDPHP.

Temperature-dependent toxicity and mechanisms of florfenicol on the embryonic development of marine medaka (Oryzias melastigma).

The extensive use of antibiotics and their persistence in the environment have seriously threatened marine ecosystems in recent years. The frequent occurrence of extreme weather due to climate change has also increased the uncertainty of effective toxicity identification and risk assessment of the chemicals of concern. This study aimed to investigate the toxic effects and potential mechanisms of florfenicol (0.5, 10, 100, 500 and 1000 μg/L) on the embryonic development of marine medaka (Oryzias melastigma) through 21-d experiment under three temperature exposure scenarios (20, 25, and 30 °C). Considering embryo development, the highest level of florfenicol at 1000 μg/L decreased the hatching success at 25 °C, whereas the total inhibition of hatching was observed at 20 °C regardless of florfenicol concentrations of concern. The ATP content was inhibited by the highest florfenicol dose at 20 °C, while stimulated ATP content at 20 °C and 30 °C, compared to 25 °C. Fluctuation of temperatures, especially at 20 ℃, induced oxidative stresses, including increased MDA contents and decreased CYP450 and HSP90 contents. For inflammatory response-related genes (nf-κb, nlrx1, pycard, caspase 1, and il-1β), an increase of florfenicol dose led to gene upregulation at 25 °C. Conversely, gene upregulation was also observed for all treatments at 30 °C, while predominantly downregulation was observed for all treatments at 20 °C. For genes related to DNA damage and apoptosis (pi3k, akt, foxo1, tp53, bcl-2, bax, apaf-1, caspase 9, and 3), alterations in gene expression were evident for all florfenicol treatments at both 20 °C and 30 °C. Therefore, this study suggests that the combined effects of florfenicol and temperature may disrupt the normal function of mitochondria, impacting ATP production, thereby inducing oxidative stress, inflammatory response, DNA damage, and apoptosis, ultimately resulting in decreased hatching rates and increased mortality. Furthermore, the key findings of this study reveal the complex interactions between florfenicol and temperature and emphasize the need to consider temperature when identifying toxicity and mechanisms, as well as the ecological risk assessment of florfenicol in marine environments.

The effects of Aronia melanocarpa (AM) dietary supplementation on production performance, meat and egg quality, yolk volatile substances and antioxidant capacity of laying hens

ABSTRACT The effects of Aronia melanocarpa (AM) dietary supplementation on production performance, meat and egg quality, yolk volatile substances and antioxidant capacity of laying hens. A total of 480 Roman brown laying hens aged 25 wks were randomly divided into control, LAM, MAM and HAM groups and fed 0%, 1%, 4% and 7% AM, respectively. The results showed that AM dietary supplementation significantly increased the eggshell strength, significantly reduced 24 h dripping loss of breast and thigh muscles, and increased pH and 48 h dripping loss of breast and thigh muscles. Cooking loss was significantly lower only in the HAM group for breast muscle. The MAM group was able to significantly increase albumen height and Haugh units. Esters gradually decreased with the increase in AM content, and the addition of AM led to a decrease in alcoholic compounds but increased the contents of alkenes, alkanes, ketones, and acids. The dietary AM supplementation significantly reduces the MDA content of serum, liver, ovary, chest muscle, thigh muscle and yolk and increases the activities of GSH-Px, T-SOD and T-AOC. In summary, dietary AM supplementation could improve antioxidant capacity and expression of related genes, thereby improving meat and egg quality.

Correlations between biochemical indicators determined in the somatosensory cortex and hippocampus and morphological indicators, functional state of the central nervous system and neurological deficit in rats

Annotation. Today, the development and clinical implementation of drugs capable of preventing and/or inhibiting the apoptosis phenomenon is considered the main direction of cytoprotection in cerebral ischemia. Among the latter, the use of citicoline attracts attention. The purpose – to establish and analyze correlations between biochemical parameters determined in the somatosensory cortex and hippocampus of rats, and morphological manifestations of neuroapoptosis and parameters of CNS functioning in animals with experimental IR (ischemia-reperfusion) of a brain. The study was carried out on 75 four-month-old male Wistar rats weighing 160-190 g. In 65 animals, the experimental model of IR was performed by placing ligatures on the internal carotid arteries on both sides under propofol anesthesia. After cerebral IR modelling, 0,9% NaCl in a dose of 2 ml/kg was injected into the femoral vein of 40 rats. A group of animals (25 rats) immediately after the start of brain reperfusion was intravenously injected with citicoline (at a dose of 250 mg/kg). The control group of animals included sham-operated rats (10 animals). Assessment of neurological deficit in rats was carried out according to the stroke-index scale of C.P. McGrow, the functional state of the central nervous system - by the "open field" method, and morphometric and biochemical indicators were studied on the 7th and 14th day after IR. Statistical processing of the research results was carried out using the computer program Statistica 7.0 (StatSoft Inc. production, USA). The methods of non-parametric statistics were used (in particular, the Wilcoxon-Mann-Whitney U-test, Spearman's non-parametric correlation rank analysis). Reliable correlations are observed between the functional indicators of the central nervous system in rats (according to the results of the "open field" test) and biochemical indicators (such as the content of lactate, glucose, MDA, SDH, SOD and NO-synthase in the tissue of the somatosensory cortex and hippocampus), which indicate a change in the motor activity of rats with enzymatic and metabolic imbalance in the brain tissue of the specified areas. In addition, there are multiple linear relationships between biochemical and morphological indicators determined in the somatosensory cortex and hippocampus, which indicates a close functional interaction of the specified brain departments with each other (observed in all groups of rats). With subtotal IR of the brain in rats, there are specific direct correlations between the protein content in brain tissue and morphological indicators, as well as neurological deficit, which are recorded in the subacute period of ischemia (the 7th day) both in the somatosensory cortex and in the hippocampus. Specific inverse correlations also occur between MDA activity in the somatosensory cortex (the 14th day after ischemia) and morphological indicators in the hippocampus. The use of citicoline as a neuroprotector immediately after IR in rats did not cause the appearance of new highly specific correlational dependencies or the disappearance of existing ones.

The Effects of Artificial vs. Natural Rearing on Growth Performance, Thyroid Hormone Levels, Locomotor Activity, Carcass Traits and Meat Quality Characteristics in Chios Lambs

Artificial rearing (AR) of lambs is nowadays a common practice in Mediterranean dairy sheep production systems to enhance the milk available for cheese or yoghurt manufacturing. The sufficient growth of lambs in an AR system is vital for the economic success of dairy sheep farms. However, AR is often associated with negative impacts on the performance and physiology of lambs. Greece is one of the major producers of ovine milk; nevertheless, data concerning the effects of artificial rearing in lambs of Greek autochthonous breeds are not available. Therefore, the present study aimed to examine the influence of artificial rearing on growth performance, thyroid hormone levels, locomotor activity, carcass traits and meat quality characteristics in lambs of the Chios breed, which is one of the most well-known Greek dairy sheep breeds. Twenty-one singleton male lambs were assigned into two feeding regimes; natural rearing NR (n = 11) and AR (n = 10). The lambs’ behavior was continuously videotaped until weaning, and their standing percentage was recorded as an activity index. Blood samples were collected from the jugular vein of lambs on days 3, 10, 17 and 40 after birth to assess thyroid hormone levels. The body weight of lambs was also recorded weekly. At the age of 45 days, lambs were fasted for 12 h, weighed and slaughtered. The weights of the carcass and internal organs were measured, while samples of the longissimus dorsi muscle were used for the determination of meat pH, color, water holding capacity, shear force and oxidative stability values. As indicated, body weight (kg) at birth was greater in NR vs. AR group and this difference was maintained till day 35 (p &lt; 0.05), although body gain (kg) was generally not significantly different between NR and AR lambs, with the exception of the first week, when NR showed a greater value compared with the AR lambs (p &lt; 0.001). On day 42, no significant differences between lamb groups for body weight were observed. Levels for triiodothyronine (T3), thyroxine (T4) and the free form of T3 (FT3) were greater, whereas the standing percentage was lower in NR compared with AR lambs (p &lt; 0.05). The feeding regime of lambs did not affect carcass traits, internal organ and fat tissue weights, except for cold carcass yield which was greater in AR vs. NR lambs. No significant differences were observed between the two lamb groups in meat quality characteristics, such as pH, color, water holding capacity and shear force values, although MDA content was decreased in AR lambs indicating an improved oxidative stability. In conclusion, artificial rearing appears to be a feasible strategy for Chios lamb meat production, since it does not negatively influence carcass traits and meat quality characteristics, while a positive effect in meat oxidative stability is observed.

Photosynthetic response of Solidago gigantea Aition and Calamagrostis epigejos L. (Roth) to complex environmental stress on heavy metal contaminated sites

Studies of in situ plant response and adaptation to complex environmental stresses, are crucial for understanding the mechanisms of formation and functioning of ecosystems of anthropogenically transformed habitats. We study short- and long-term responses of photosynthetic apparatus (PSA) and anti-oxidant capacity to complex abiotic stresses of common plants Calamagrostis epigejos and Solidago gigantea in semi-natural (C) and heavy metal contaminated habitats (LZ). We found significant differences in leaf pigment content between both plant species growing on LZ plots and their respective C populations. The average values of leaf chlorophyll indexes were 27% lower in the LZ populations of both species and significantly lower in Sg plants in comparison to Ce ones. The average values of the anthocyanin index in CeLZ and SgLZ populations were significantly higher (by 18%) than in their respective controls. In both Ce and Sg plants occurring on LZ plots, the average leaf flavonol indexes were higher than on their controls by 31% and 15% and this index was significantly higher in SgLZ population than CeLZ and CeC plants (by 34% and 54%, respectively). Both Ce and Sg populations growing on LZ plots showed significantly lower photosynthetic rate (A), transpiration rate (E) and stomatal conductance (gs) in comparison to controls. On the other hand, a significantly higher photosynthetic rate was detected in SgLZ than in CeLZ populations. The catalase activities were significantly higher than recorded in Sg than in Ce tissues, irrespective of the plot type. They were also higher in LZ populations than those in controls for both species. Moreover, the H2O2 content in Sg tissues was significantly higher than those in Ce. Hydrogen peroxide content in CeLZ and SgLZ were respectively 39% (non-significant) and 57% higher, compared to their controls. The reverse pattern was found in the case of MDA, whose concentration was significantly higher in the leaves of Ce population compared to the control population. The average MDA concentration in CeLZ populations was 17% higher than in the CeC. In the case of Sg no significant differences were found. Mechanisms of plant species adaptation to industrial areas are crucial for species selection and planning effective reclamation of them. The analysis of chlorophyll fluorescence induction curves as well as well as the results of JIP test revealed the decreased of Fj value despite positive ΔK–band in SgLZ and CeLZ plants suggesting the increased rate of electron transfer from QA to QB at the acceptor side of PSII, thus a high quantity of P680+ and/or effective quenching by exogenous molecules. The increase in the I–P part of the induction curve typically attributed to the reduction of electron transporters (ferredoxin, intermediary acceptors, and NADP) of the PSI acceptor side was observed in both SgC and SgLZ but not in CeLZ populations. These changes demonstrate species-specific effects on electron transport during the light phase of photosynthesis under complex environmental stress. Our results show that Sg and Ce individuals developed a range of structural and functional adaptations to protect PSA against complex environmental stresses (possible combination of heavy metals, water deficiency, temperature, nutrient deficiency and salinity). Both species from LZ plots could tolerate high levels of Cd, Zn and Pb in leaf tissues. Therefore they can be potential candidates for use in phytoremediation of HM contaminated areas. However, further long-term field and experimental research on plant traits response and adaptation to complex environmental stresses on industrial habitats are needed.

Effects of Medium- and Short-Chain Fatty Acids on Growth Performance, Nutrient Digestibility, Gut Microbiota and Immune Function in Weaned Piglets

The aim of this study was to investigate the combination effects of α-glycerol monolaurate (GML) and glyceryl tributyrate (TB) on growth performance, nutrient digestibility, gut microbiota, and immune function in weaned piglets. A total of 120 weaned piglets with an average body weight (BW) of 6.88 kg were randomly allocated to one of the three dietary treatments: (1) CON: a basal diet; (2) 0.1%: a basal diet with 0.1% MSCFA (GML/TB = 1:1); (3) 0.2%: a basal diet with 0.2% MSCFA (GML/TB = 1:1). The experiment lasted 28 days. There were no differences on average daily growth (ADG), average daily feed intake (ADFI), and feed conversion ratio (FCR). Supplementation with 0.1% MSCFA increased apparent total tract digestibility (ATTD) of crude protein (CP) and gross energy (GE, p &lt; 0.05) on d 14 and increased GE (p &lt; 0.05) on d 28 compared with the CON group. The ATTD of dry matter (DM), organic matter (OM) and crude protein (CP) of piglets supplemented with 0.1% MSCFA was higher (p &lt; 0.05). Compared with the CON group, supplementation with 0.1% MSCFA increased immunoglobulin M (IgM) concentration, decreased interleukin-6 (IL-6) content (p &lt; 0.05) on d 14 and decreased malonaldehyde (MDA), interleukin-1beta (IL-1β), IL-6 concentrations (p &lt; 0.05) on d 28. Supplementation with 0.1% MSCFA increased total antioxidant capacity (T-AOC) concentration (p &lt; 0.05), decreased GSH-Px, MDA content (p &lt; 0.05) in jejunum compared with the CON group. Moreover, supplementation with MSCFA increased the activity of duodenal lipase (p &lt; 0.05) and the abundance of firmicutes and decreased the abundance of proteobacteria compared with the CON group. Overall, supplementation with MSCFA can improve nutrient digestibility, enhance immunity and antioxidant capacity, and improve the intestinal health of piglets. The combined use of MSCFA is a nutrition regulation strategy worthy of further exploration in modern animal husbandry.

Role of biotin carboxyl carrier protein subunit 2 (BCCP2) in resistance to multiple stresses in Arabidopsis thaliana.

Abiotic stresses, including drought, salinity, and temperature extremes, are serious constraints to plant growth and agricultural development. These stresses that plants face in nature are often multiple and complex. Biotin carboxyl carrier protein subunit 2 (BCCP2) is one of the two subunits of biotin carboxyl carrier protein, which is a functional subunit of acetyl coenzyme A carboxylase, primarily studied for its role in fatty acid synthesis. In this study, we identified the expression pattern of AtBCCP2 under various stress conditions, including 4mM NaHCO₃, 2mM Na₂CO₃, 150mM NaCl, 300mM D-mannitol, 100μM ABA, 5mMH₂O₂, 4°C, and 37°C. It was determined that AtBCCP2 is positively regulated by NaHCO₃, Na₂CO₃, NaCl, and ABA, but negatively regulated by D-mannitol. Phenotypic experiment confirmed that the AtBCCP2 transgenic overexpression plants exhibited increased resistance to NaHCO₃, Na₂CO₃, NaCl, and ABA stresses, but more sensitive to drought stress simulated by D-mannitol. In contrast, mutant plants showed the opposite phenotypes. Additionally, AtBCCP2 transgenic overexpression plants demonstrated stronger antioxidant activity and lower MDA content under stresses such as NaHCO₃, Na₂CO₃, NaCl, and ABA, in contrast to the mutant plants. In response to D-mannitol simulated drought stress, AtBCCP2 transgenic overexpression plants showed lower antioxidant activity and higher MDA content, while mutant plants exhibited the opposite trend. In conclusion, this study provides a theoretical basis for understanding the role of AtBCCP2 in response to multiple stresses and contributes a new gene to the pool of those involved in abiotic stress responses.