Metal-Organic Framework-Based Surface-Enhanced Raman Scattering Sensing Platform for Trace Malondialdehyde Detection in Tears.

Disease biomarkers in tears are crucial for clinical diagnosis and health monitoring. However, the limited volume of tear samples, low concentration of tear biomarkers, and complex tear composition present challenges for precise testing. We introduce a spot-on testing platform of metal-organic framework (MOF)-based surface-enhanced Raman scattering (SERS) capillary column, which is capable of target molecules selective separation and enrichment for tear biomarkers in situ detection. It consists of Au nanostars for effective SERS signal and a porous MOF shell for separating impurities through molecular sieving effect. This platform allows for simultaneous collection and detection of tear, capturing the disease biomarker malondialdehyde in tears with a 9.38 × 10-9 mol/L limit of detection. Moreover, we designed a hand-held device based on this tubular SERS sensor, successfully diagnosing patients with dry eye disease. This functional capillary column enables noninvasive and rapid diagnosis of biomarkers in biofluids, providing potential for disease diagnosis and healthcare monitoring.

Probiotics from kefir: Evaluating their immunostimulant and antioxidant potential in the carpet shell clam (Ruditapesdecussatus).

This study aimed to investigate the impact of incorporating kefir into the diet on biometric parameters, as well as the immune and antioxidant responses of the carpet shell clam (Ruditapes decussatus) after an experimental infection by Vibrio alginolyticus. Clams were divided into a control group and a treated group. The control group was fed on spirulina (Arthrospira platensis) alone. While, the treated group was fed on spirulina supplemented with 10% dried kefir. After 21 days, clams were immersed in a suspension of V. alginolyticus 5 × 105 CFU mL -1 for 30 min. Seven days after experimental infection, survival was 100% in both groups. The obtained results showed a slight increase in weight and condition index in clams fed with kefir-supplemented diet for 21 days compared to control clams. Regarding antioxidant responses, the treated group showed higher superoxide dismutase activity compared to the control group. However, the malondialdehyde level was lower in the treated clams than in the control. In terms of immune parameters, the treated group showed slightly elevated activities of phenoloxidase, lysozyme and alkaline phosphatase, whereas a decreased lectin activity was observed compared to the control group. The obtained results suggest that kefir enhanced both the antioxidant and immune response of infected clams.

Effects of hydroxytyrosol on post-thaw quality of rooster sperm.

Semen cryopreservation is one of the most important reproduction techniques in the livestock and poultry industry. Cryopreservation induces cold stress, generating reactive oxygen species (ROS) and oxidative stress causing structural and biochemical damages in sperm. In this study, we evaluated the effects of the hydroxytyrosol (HT), as an antioxidant, at the concentrations of 0, 25, 50, and 100 µg/mL on post-thaw semen quality metrics in rooster. Semen samples were collected twice a week from 10 roosters (29 weeks), processed and frozen according to experimental groups. Different quality parameters, including total motility, progressive motility, viability, morphology, membrane integrity, and malondialdehyde were measured after thawing. Results showed that 25 and 50 µg/mL of HT produced the highest percentage of total motility (51.01 ± 2.19 and 50.15 ± 2.19, respectively) and progressive motility (35.74 ± 1.34 and 35.15 ± 1.34, respectively), membrane integrity (48.00 ± 2.18 and 46.75 ± 2.18, respectively) as well as viability (53.00 ± 2.17 and 52.50 ± 2.17, respectively) compared with the other groups (p < .05). The group with 25 µg/mL of HT showed the lowest significant (p < .05) MDA concentration (1.81 ± 0.25). Our results showed that the effect of HT was not dose-dependent and optimum concentration of HT could improve functional parameters of rooster sperm after freezing-thawing. These findings suggest that HT may have protective effects on the rooster sperm during the freezing-thawing process.

A Cross-Sectional Study Comparing Oxidative Stress in Patients with Epilepsy Treated with Old and New Generation Antiseizure Medications.

Background and Objectives: Oxidative stress resulting from a disturbance of the endogenous redox system is suspected in numerous diseases of the central nervous system, including epilepsy. In addition, antiseizure medications (ASMs), especially those of the old generation, may further increase oxidative stress. To evaluate the effects of ASM generation on oxidative stress, we conducted a cross-sectional study in patients with epilepsy treated with old, new, and polytherapy. Materials and Methods: The antioxidant activity of superoxide dismutase, catalase, glutathione peroxidase, and glutathione reductase, as well as the concentrations of malondialdehyde, protein carbonyl, nitrate, nitrite, and glutathione in reduced and oxidized forms, were measured in 49 patients with epilepsy and 14 healthy controls. In addition, the plasma concentrations of ASMs and metabolites of carbamazepine and valproic acid were measured in the patients. Results: Patients with epilepsy showed increased activities of superoxide dismutase and catalase (p < 0.001), concentrations of glutathione disulfide and markers of nitric oxide metabolism (p < 0.001), and decreased activities of glutathione peroxidase, glutathione reductase, glutathione, and nitrite concentrations (p ≤ 0.005) compared to healthy controls. A comparison of ASM generations revealed increased levels of superoxide dismutase and catalase (p ≤ 0.007) and decreased levels of glutathione peroxidase and glutathione reductase (p ≤ 0.01) in patients treated with old ASMs compared to those treated with new generation ASMs. In addition, an increase in protein carbonyl and nitric oxide metabolites (p ≤ 0.002) was observed in patients treated with old generation ASMs compared to those treated with new generation ASMs. Most oxidative stress parameters in patients receiving polytherapy with ASMs were intermediate between the results of patients treated with the old and new generations of ASMs. Conclusions: An increase in oxidative stress markers and modulation of antioxidant enzyme activities was observed in patients with epilepsy compared to controls. The results of our study showed significantly higher oxidative stress in patients treated with old ASMs compared to those treated with new generation ASMs.

Salivary lipid changes in young adult tobacco smokers and e-cigarette users: a hidden risk to oral health?

STUDY DESIGN: A cross-sectional, age- and gender-matched study was conducted to investigate the effects of different forms of nicotine delivery on salivary lipid profiles among young adult novice smokers compared to non-smokers. OBJECTIVE: To assess the effect of smoking traditional cigarettes, e-cigarettes, and heated tobacco products (HTPs) on the levels of specific sphingolipids (sphingosine, sphinganine, and sphingosine-1-phosphate), various ceramides, and lipid peroxidation products [malondialdehyde (MDA) and 4-hydroxynonenal (4-HNE)] in both unstimulated and stimulated saliva samples collected from healthy young adults who had been smoking for 1-3 years and used only 1 of the 3 nicotine delivering methods. METHODS: Selection criteria included healthy young adults under 30 years old, with normal BMI and typical diet composition, and with no oral inflammatory lesions, orthodontic/dental appliances, or recent intake of medications or supplements. A total of 75 smokers and 25 non-smokers were enrolled in the study. Smokers were categorized into three groups, each comprising 25 individuals: traditional cigarette smokers, e-cigarette users, and HTPs smokers. Saliva samples were collected and analyzed for sphingolipid concentrations using ultra-high-performance liquid chromatography-tandem mass spectrometry. The concentrations of MDA and 4-HNE were measured using colorimetric and ELISA assays, respectively. RESULTS: The average smoking intensity in the traditional cigarette group was 10 cigarettes per day. Salivary sphingolipid and ceramides concentrations were significantly lower in smokers compared to non-smokers across all nicotine delivery methods (p < 0.0001). Moreover, traditional cigarette smokers exhibited higher levels of 4-HNE and MDA in both stimulated and unstimulated saliva, compared to non-smokers (p < 0.01). In stimulated saliva, both MDA and 4-HNE in e-cigarette users, and MDA in HTPs users, showed significantly lower concentrations than their comparators in traditional cigarette smokers (p < 0.01). CONCLUSION: Different nicotine delivery methods impact salivary lipid profile during the initial period of smoking habit. Reduced sphingolipids and elevated lipid peroxidation products suggest a disturbed lipid balance in the oral cavity due to enhanced oxidative stress within the salivary glands of novice smokers.

Oxidative stress-related biomarkers in chronic periodontitis patients with or without type 2 diabetes: A systematic review and meta-analysis.

OBJECTIVE: The purpose of this meta-analysis was to look at the differences in oxidative stress (OS) biomarkers between type 2 diabetes mellitus with chronic periodontitis (DMCP) and chronic periodontitis (CP) patients. BACKGROUND: Oxidative stress has been shown to be a key pathogenic component in DMCP. However, it is unclear whether oxidative stress levels differ in periodontitis patients with or without diabetes. METHOD: A systematic search was conducted on PubMed, Cochrane, and Embase databases. Studies of DMCP participants were used as the experimental group and CP participants were used as the control group. Results are expressed as mean effects. RESULTS: Of a total of 1989 articles, 19 met the inclusion criteria. We found the levels of catalase (CAT) levels were reduced in the DMCP group compared with the CP group. However, there was no significant difference in the levels of superoxide dismutase (SOD), total antioxidant capacity (TAOC) malondialdehyde (MDA), and glutathione (GSH) between the two groups. And high heterogeneity was observed in some of the studies evaluated. CONCLUSION: Despite the limitations of this study, our results support the theory that there is an association between T2DM and the levels of OS-related biomarkers, especially CAT, in CP subjects, suggesting that OS plays an important role in the pathogenesis and development of DMCP.

Transition cow clusters with distinctive antioxidant ability and their relation to performance and metabolic status in early lactation.

Metabolic and oxidative stress have been characterized as risk factors during the transition period from pregnancy to lactation. Although mutual relations between both types of stress have been suggested, they rarely have been studied concomitantly. For this, a total of 99 individual transition dairy cows (117 cases, 18 cows sampled during 2 consecutive lactations) were included in this experiment. Blood samples were taken at -7, 3, 6, 9, and 21 d relative to calving and concentrations of metabolic parameters (glucose, ß-hydroxybutyric acid (BHBA), nonesterified fatty acids, insulin, insulin-like growth factor 1, and fructosamine) were determined. In the blood samples of d 21, biochemical profiles related to liver function and parameters related to oxidative status were determined. First, cases were allocated to 2 different BHBA groups (ketotic vs. nonketotic, N:n = 20:33) consisting of animals with an average postpartum BHBA concentration and at least 2 out of 4 postpartum sampling points exceeding 1.2 mmol/L or remaining below 0.8 mmol/L, respectively. Second, oxidative parameters [proportion of oxidized glutathione to total glutathione in red blood cells (%)], activity of glutathione peroxidase, and of superoxide dismutase, concentrations of malondialdehyde and oxygen radical absorbance capacity were used to perform a fuzzy C-means clustering. From this, 2 groups were obtained [i.e., lower antioxidant ability (LAA80%, n = 31) and higher antioxidant ability (HAA80%, n = 19)], with 80% referring to the cutoff value for cluster membership. Increased concentrations of malondialdehyde, decreased superoxide dismutase activity, and impaired oxygen radical absorbance capacity were observed in the ketotic group compared with the nonketotic group, and inversely, the LAA80% group showed increased concentrations of BHBA. In addition, the concentration of aspartate transaminase was higher in the LAA80% group compared with the HAA80% group. Both the ketotic and LAA80% groups showed lower dry matter intake. However, a lower milk yield was observed in the LAA80% group but not in the ketotic group. Only 1 out of 19 (5.3%) and 3 out of 31 (9.7%) cases from the HAA80% and LAA80% clusters belong to the ketotic and nonketotic group, respectively. These findings suggested that dairy cows vary in oxidative status at the beginning of the lactation, and fuzzy C-means clustering allows to classify observations with distinctive oxidative status. Dairy cows with higher antioxidant capacity in early lactation rarely develop ketosis.

The evaluation of oxidative stress and inflammation markers in serum and saliva of the patients with temporomandibular disorders.

Background/aim: Temporomandibular Disorders (TMD), as in the occurrence of many diseases, have been associated with oxidative stress (OS) resulting from the disruption of antioxidant mechanisms and the accumulation of reactive oxygen species in tissues. This study was designed to compare salivary and serum OS and inflammation markers of individuals with TMD and healthy subjects. Materials and methods: A prospective cross-sectional study was conducted. Twenty-seven TMD patients diagnosed with disc displacement (DD) according to Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD) and 17 healthy subjects were enrolled in the study. Prior to any treatment, serum, and saliva samples were taken from the patients and centrifuged, and stored at -80 °C until analyzed. All samples were examined for Interleukin-6 (IL-6), Malondialdehyde (MDA), and 8-hydroxydeoxyguanosine (8-OHdG) concentrations. Results: There was no significant difference between the groups regarding median values of 8-OHdG, IL-6, and MDA (p > 0.05). When the relationship between serum and salivary 8-OHdG, IL-6, and MDA levels in all subjects was evaluated, there was a strong positive correlation between the levels of 8-OHdG and IL-6 in the serum (r = 0.752, p <0.001). In the study group, when the relationship between pain levels and serum and saliva 8-OHdG, IL-6, and MDA levels was assessed, a positive and strong correlation was found between the levels of 8-OHdG and IL-6 in serum. Conclusion: Although the strong correlation between pain scores and serum 8-OHdG and MDA levels supports the hypothesis that inflammation and OS mechanisms may be interrelated, according to the results of the study, inflammatory and OS markers in patients with TMD were not different from healthy individuals.

Effect of chelated source of additional zinc and selenium on performance, yolk fatty acid composition, and oxidative stability in laying hens fed with oxidised oil.

1. The following study determined whether the effects of the combined addition of zinc amino acid complex (ZA) and selenomethionine (SM) was superior to their single addition in controlling the oxidative stress induced by dietary oxidised fat in laying hens.2. Two hundred and forty 32-week-old laying hens were divided into the following dietary treatments (each consisting of six replicates of eight birds): 1) a fresh soy oil (FSO) diet; 2) an oxidised soy oil (OSO) diet; 3) an OSO diet plus 20 mg zinc as ZA/kg (OSO+ZA); 4) an OSO diet plus 0.2 mg selenium as SM/kg (OSO+SM); and 5) an OSO diet plus ZA and SM (OSO+ZA+SM).3. After 10 weeks of feeding hens, feed intake, egg production, and egg mass in the OSO+ZA+SM group were similar to the FSO group but better (P < 0.05) than those in the OSO group. Shell thickness and shell breaking strength were significantly improved by the OSO+ZA and OSO+ZA+SM treatments.4. Increases in the yolk concentrations of palmitic acid and total saturated fatty acids (SFA), and decreases in yolk linoleic acid, n-6 polyunsaturated fatty acids (PUFA), total PUFA, and PUFA/SFA ratio were induced by dietary oxidised fat which were normalised (P < 0.05) by OSO+SM and OSO+ZA+SM.5. An increase (P < 0.05) in malondialdehyde and a decrease in 2,2-diphenyl-picrylhydrazyl radical scavenging activity in the yolk, induced by dietary oxidised fat, was significantly improved by all dietary supplementations, but only birds fed the OSO+ZA+SM diet exhibited similar values to those fed FSO.6. In conclusion, the simultaneous inclusion of organic zinc plus selenium in the oxidised fat diets was beneficial for improving egg-laying performance, yolk fatty acid profile, and oxidative stability, but not for internal egg quality, compared with either zinc or selenium alone in laying hens.

[Components and lipid-lowering effect of total saponins from underground part of Gynostemma pentaphyllum].

The saponins in different parts of Gynostemma pentaphyllum were analyzed via UPLC-Q-TOF-MS~E. A total of 46 saponins were identified, and the underground part had 26 saponins more than the aboveground part, most of which were trisaccharide saponins. The rat model of hyperlipidemia was established with high-fat diet. This study explored the lipid-lowering activity of total saponins in the underground part of G. pentaphyllum, so as to provide a theoretical basis for the comprehensive utilization of the underground part of G. pentaphyllum. A total of 99 healthy SD rats were randomly assigned into a blank group, a model group, a positive drug group, an aboveground total saponins group, and low-, medium-, and high-dose underground total saponins groups. Except the blank group, the other groups were fed with high-fat diet for 6 weeks. Then, the blood was collected from the orbital cavity to determine whether the modeling was successful according to the serum levels of total cholesterol(TC) and triglyceride(TG). After intragastric administration of the corresponding agents for 30 continuous days, the physical state of the rats were observed, and the body weight and liver specific gravity were measured. Furthermore, the levels of TC, TG, low-density lipoprotein cholesterol(LDL-C), high-density lipoprotein cholesterol(HDL-C), alanine transaminase(ALT), aspartate transaminase(AST), bilirubin, and total bile acids in serum, as well as the levels of superoxide dismutase(SOD), malondialdehyde(MDA), peroxidase proliferator-activated receptor(PPAR-γ) in the liver tissue, were determined. The pathological changes of liver was observed via HE staining. The results showed that the aboveground total saponins and medium-and high-dose underground total saponins can treat hepatocyte steatosis, lower TC, TG, LDL-C, ALT, AST, total bilirubin, MDA, and PPAR-γ levels, and increase HDL-C and SOD levels in the model rats. The effect tended to be more obvious with the increase in dosage. Therefore, the total saponins in the underground part of G. pentaphyllum have good pharmacological effect of reducing blood lipid, which provides a theoretical basis for the comprehensive utilization of the underground part of G. pentaphyllum.

Development and validation of a methodology for quantitative determination of malondialdehyde by HPLC-MC/MS.

A bioanalytical technique for quantitative determination of MDA by HPLC-MS/MS. The proposed method for determining MDA includes the release stage of bound MDA and excludes the derivatization reaction. The lower limit of quantitative detection was 600 nmol/l, the volume of the required sample was 10 µl, the analysis time was 7 min. The range of concentrations obtained during the study makes it possible to use this bioanalytical technique to determine the concentration of MDA in biological material when assessing physiological and pathological conditions.

Malondialdehyde, an Oxidative Stress Marker in Oral Squamous Cell Carcinoma-A Systematic Review and Meta-Analysis.

OBJECTIVE: To qualitative and quantitatively review published literature assessing the oxidative stress marker malondialdehyde (MDA) in oral squamous cell carcinoma (OSCC). METHODOLOGY: Pubmed (MeSH), Science Direct, Scopus, Web of Science, Willey Online Library, Cochrane, and Cross Reference were searched for studies assessing MDA levels in OSCC samples. RESULTS: From the 1008 articles identified, 849 were excluded based on title and abstract screening due to duplication and irrelevance to the topic of interest. Full-text assessment of the remaining 159 articles led to the inclusion of only 46 articles that satisfied the selection criteria. Of these, only 26 studies had data compatible for quantitative analysis. The MDA levels in OSCC groups are significantly increased (p < 0.00001) in plasma, serum, and saliva samples in the majority of the studies evaluated. In contrast, MDA levels in OSCC tissue samples are significantly attenuated (p < 0.00001) compared to healthy controls, supported by fewer studies. CONCLUSIONS: The augmented MDA levels in plasma, serum, and saliva samples of the OSCC reflect the heightened oxidative stress level accurately. Further studies are required to understand the attenuated MDA levels in the tissue samples of OSCC. Correlation analysis between MDA levels with established clinicopathological prognostic markers could aid in formulating oxidative stress-based prognostication and treatment planning.

The rainbow protocol: A sequential method for quantifying pigments, sugars, free amino acids, phenolics, flavonoids and MDA from a small amount of sample.

The elucidation of plant health status requires quantifying multiple molecular metabolism markers. Until now, the extraction of these biomarkers is performed independently, with different extractions and protocols. This approach is inefficient, since it increases laboratory time, amount of sample, and could introduce biases or difficulties when comparing data. To limit these drawbacks, we introduce a versatile protocol for quantifying seven of the most commonly analysed biomarkers (photosynthetic pigments, free amino acids, soluble sugars, starch, phenolic compounds, flavonoids and malondialdehyde) covering substantial parts of plant metabolism, requiring only a minimum sample amount and common laboratory instrumentation. The procedures of this protocol rely on classic methods that have been updated to allow their sequential use, increasing reproducibility, sensibility and easiness to obtain quantitative results. Our method has been tested and validated over an extended diversity of organisms (Arabidopsis thaliana, Solanum lycopersicum, Olea europaea, Quercus ilex, Pinus pinaster and Chlamydomonas reinhardtii), tissues (leaves, roots and seeds) and stresses (cold, drought, heat, ultraviolet B and nutrient deficiency). Its application will allow increasing the number of parameters that can be monitored at once while decreasing sample handling and consequently, increasing the capacity of the laboratory.

Possible protective effect of TNF-α inhibition and triad NO/cGMP/VEGF activation on gastric ulcer in rats.

Peptic ulcers are one of the world's major gastrointestinal disorders, embracing both gastric and duodenal ulcers, and affecting 10% of the world population. The current study aimed to investigate the possible protective effect of tadalafil and pentoxifylline (PTX) on indomethacin-induced peptic ulcers. Male albino rats were divided into five groups: control group; ulcerated group; Indomethacin + Tadalafil, in which animals were pretreated with tadalafil orally before indomethacin; Indomethacin+ PTX, in which animals were pretreated with PTX orally before indomethacin; and Indomethacin + Tadafil + PTX. Indomethacin treatment revealed histopathological changes and ulcer scoring and ulcer index were markedly increased. Serum levels of prostaglandin and heme oxygenase-1 were significantly decreased. The ulcerogenic also induced marked oxidative stress as evident from the increased malondialdehyde, decreased in gastric glutathione content and superoxide dismutase activity, while the gastric myeloperoxidase was increased. Gastric nitric oxide content was decreased and the expression of vascular endothelial growth factor was downregulated while the tumor necrosis factor α (TNF-α) level was dramatically increased. Pretreatment of the ulcerative group by either tadalafil or PTX or their combination improved all these pathological changes. Tadalafil or PTX may have a role in protecting gastric mucosa damage caused by indomethacin which may be useful in the future for the treatment of gastric ulceration.

Correlations of Silent Information Regulator of Transcription 1 (SIRT1) Expression, Inflammatory Factors, and Oxidative Stress with Pulmonary Function in Patients with Acute Exacerbation of Chronic Obstructive Pulmonary Disease (AECOPD).

BACKGROUND The aim of this study was to investigate the correlations of silent information regulator of transcription 1 (SIRT1) expression, inflammatory factors, and oxidative stress with pulmonary function in patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD). MATERIAL AND METHODS Bronchoalveolar lavage fluid (BALF) was collected from 188 patients with COPD (83 in stable phase and 105 in acute exacerbation phase) and 56 healthy controls. Subsequently, the SIRT1 expression levels, the IL-6 and IL-8 levels (the representatives of inflammatory factors), and the MDA and SOD levels (indicative of oxidative stress) were detected via enzyme-linked immunosorbent assay. Correlations of SIRT1 expression, inflammatory factors, and oxidative stress with pulmonary function parameters [forced expiratory volume in one second (FEV1)/forced vital capacity (FVC) and FEV1] were measured via Spearman's correlation analysis. RESULTS The levels of inflammatory factors and oxidative stress were elevated and SIRT1 expression remarkably declined in patients with AECOPD compared with those in healthy controls and stable COPD patients (P<0.05). Spearman's correlation analysis revealed that SIRT1 expression, interleukin (IL)-6, and IL-8 were strongly associated with pulmonary function parameters (FEV1/FVC and FEV1) in patients with AECOPD (P<0.001), while no such obvious correlation was observed in stable COPD patients. CONCLUSIONS Oxidative stress and expression levels of inflammatory factors are evidently elevated and SIRT1 expression declines in patients with AECOPD. Moreover, SIRT1 expression is positively associated with pulmonary function parameters, while IL-6 and IL-8 exhibit negative correlations with pulmonary function parameters.

Simultaneous determination of cellular adenosine nucleotides, malondialdehyde, and uric acid using HPLC.

Adenine nucleotides and malondialdehyde (MDA) are key components involved in energy metabolism and reactive oxygen species (ROS) production. Measuring the levels of these components at the same time would be critical in studying mitochondrial functions. We have established a HPLC method to simultaneously measure adenosine triphosphate, adenosine diphosphate, adenosine monophosphate, MDA, and uric acid (UA). The samples were treated with perchloric acid followed by centrifugation. After neutralization, the supernatant was subjected to HPLC determination. HPLC was performed using a C18 chromatographic column, isocratic elusion, and UV detection. The detection and quantification limits for these components were determined with standard solutions. The precision, repeatability, and 24-h stability were evaluated using cellular samples, and their relative standard deviations were all within 2%. The reproducibility and efficiency were confirmed with sample recovery tests and the observed oxidative effects of H2 O2 on Jurkat cells. With this method, we discovered the dependence of energy and oxidative states on the density of Jurkat cells cultured in suspension. We also found a significant correlation between UA in serum and that in saliva. These results indicate that this method has good accuracy and applicability. It can be used in biological, pharmacological, and clinical studies, especially those involving mitochondria, ROS, and purinergic signaling.

Oridonin induces ferroptosis by inhibiting gamma-glutamyl cycle in TE1 cells.

Oridonin (Ori) is a natural tetracyclic diterpenoid active compound with excellent antitumor activity, but the mechanism of Ori on esophageal cancer cell, TE1, remains unclear. In this study, we examined the levels of intracellular iron, malondialdehyde, and reactive oxygen species after Ori treatment, while interfering with the effects of Ori with ferroptosis inhibitor, demonstrating that Ori's inhibition of TE1 cell proliferation is associated with ferroptosis. To understand the molecular mechanism of Ori, we performed UPLC-MS/MS metabolomics profiling on TE1 cells, which show that gamma-glutamyl amino acids (gamma-glutamylleucine, gamma-glutamylvaline), 5-oxoproline, glutamate, GSH, and GSSG are changed significantly after Ori treatment. Meanwhile, the activity of gamma-glutamyl transpeptidase 1 (GGT1) decreased. This revealed that Ori inhibited the gamma-glutamyl cycle in TE1 cells. Furthermore, we found that Ori can covalently bind to cysteine to form the conjugate oridonin-cysteine (Ori-Cys), resulting in the inhibition of glutathione synthesis, which is consistent with the decrease in the enzymatic activity of glutamate cysteine ligase catalytic subunit (GCLC). Eventually, the value of intracellular GSH/GSSG was reduced, and the enzymatic activity of the glutathione peroxidase 4 (GPX4) was significantly decreased. In conclusion, our experiments indicated that Ori can inhibit the gamma-glutamyl cycle, thereby inducing ferroptosis to exert anti-cancer activity.

Milk metabolites, proteins and oxidative stress markers in dairy cows suffering from Staphylococcus aureus subclinical mastitis with or without spontaneous cure.

Our objective was to evaluate relationships between milk components (acute phase proteins, enzymes, metabolic parameters and oxidative indices) and the spontaneous cure outcome of Staphylococcus aureus subclinical mastitis in dairy cows. The values of haptoglobin, serum amyloid A (SAA), malondialdehyde (MDA), total antioxidant capacity, milk urea nitrogen (MUN), lactate dehydrogenase (LDH), alkaline phosphatase (ALP), electrolytes (Cl and K), total protein, albumin, α-lactalbumin, ß-lactoglobulin, and immunoglobulin were measured in milk samples of S. aureus subclinical mastitis cows with spontaneous cure (n = 23), S. aureus subclinical mastitis cows without spontaneous cure (n = 29) and healthy cows (n = 23). The comparison of measured parameters revealed that subclinical mastitis cows with spontaneous cure had lower ALP and haptoglobin concentrations both at diagnosis and after cure (P < 0.05). In contrast, total antioxidant capacity and MDA concentration in subclinical mastitis cows without spontaneous cure significantly increased with time (P < 0.05). We can suggest that elevated haptoglobin concentration and higher ALP activity indicative of enhanced oxidative stress could potentially serve as early diagnostic indicators of chronic disease and the persistence of S. aureus subclinical mastitis in dairy cows.

Chemical Characteristics and Oxidative Stability of Buffalo Mozzarella Cheese Produced with Fresh and Frozen Curd.

Milk and dairy products can have variable contents of antioxidant compounds that contribute to counteract the oxidation of lipids and proteins during processing and storage. The content of active antioxidant compounds is closely linked to their protection by oxidation. Freezing is one of the factors that can reduce antioxidant activity. Freezing of milk or curd is frequently used in case of the seasonality of milk production and/or seasonal increased demand for some products. In this paper, the effect of using frozen curd on the oxidative stability of buffalo Mozzarella cheese was evaluated. Samples of buffalo Mozzarella with different frozen curd content (0%, 5%, 20%, and 50%) were produced and analyzed at one and nine days. Mozzarella cheese with higher frozen curd content had a significant increase in redox potential parallel to the decrease in antioxidant activity, showing less protection from oxidation. Lipid and protein oxidation, expressed respectively by malondialdehyde and carbonyl content, increased significantly with increasing frozen curd. At nine days, carbonyls significantly increased while malondialdehyde content did not vary, showing that during storage, fat was more protected from oxidation than protein. The average carbonyl levels were comparable to those of some cooked cheeses, and the malondialdehyde levels were even lower. The results of this study stimulate the investigation of new strategies to decrease the oxidative damage in cheeses produced in the presence of factors decreasing oxidative stability.

Evaluation of Wound-Healing and Antioxidant Effects of Marantodes pumilum (Blume) Kuntze in an Excision Wound Model.

Marantodes pumilum (MP) is a great source of herbal medicine used traditionally by both men and women for various purposes. MP may have potential wound-healing effects due to its diverse biological properties. An extensive study was conducted in a normal male rat model for determining the effects of MP var. pumila (MPvp) and var. alata (MPva) on the wound healing process. Here, 126 male Sprague-Dawley rats were divided randomly into seven groups as follows: sham-operated (SH), vehicle dressing (VD), flavine dressing (FD), MPvp leaves (PL), MPvp roots (PR), MPva leaves (AL), and MPva roots (AR). The parameters studied were the percentage of wound contraction, histomorphology study by hematoxylin and eosin (H&E), Masson-Goldner trichrome (MGT), and immunohistochemistry (IHC) staining. In addition, the levels of enzymatic antioxidants and malondialdehyde were also measured in the wound tissue homogenates. Wounds treated with extracts (PL, PR, AL, and AR) showed significantly faster healing (p < 0.05) compared to untreated and control groups (SH, VD, and FD). Histological analysis among MP-treated groups revealed better re-epithelialization, higher collagen deposition, enhanced fibronectin content and fibroblast cells, and higher fiber transformation from collagen-III to collagen-I, accompanied with a significant surge in enzymatic antioxidant activities and a decline in lipid peroxidation. MP has antioxidant effects that may enhance wound healing in the rat model.