Thiobarbituric Acid Reactive Substances Assay Research Articles

Sorption Isotherm and Effect of Packaging Materials on Proximate, Organoleptic and Lipid Oxidation Quality of Chicken Nuggets Stored at Two Different Temperatures

The study Investigated sorption isotherm features and the effects of packaging materials of both high-density and low-density polyethylene on sensory properties, proximate composition and lipid oxidation of chicken nuggets in storage. Boneless chicken breast was procured from a reputable source and cut into sizeable weight of 50g each to produce the chicken nuggets used for the studies. The experimental setup for sorption isotherm analysis in this study involved using a gravimetric method to equilibrate the product at various relative humidity levels. Five (5g) grams of oven-dried chicken nugget samples of known moisture content were placed in a controlled humidity chamber, and the weight changes of the samples were monitored until equilibrium was reached. GAB equation was used to evaluate the product ware activity and monolayer moisture. Sensory assessment was evaluated using a 9- point hedonic scale while proximate composition of the chicken nuggets samples was determined according to the methods of the Association of Official Analytical Chemists. Thiobarbituric acid reactive substances (TBARS) assay was used to measure malondialdehyde (MDA) levels as a marker of lipid oxidation. The investigation showed that sensory qualities, proximate composition and lipid oxidation of chicken nuggets were markedly affected by the variance in temperature settings, with distinct disparities seen among different materials for packaging. Chicken nuggets stored at 25 0C caused a significant loss in organoleptic properties and some nutritional composition such as protein and fat. Storage of chicken nugget at 25 0C and 40 C showed a varied values in malondialdehyde as content ranged from 0.52 to 0.81 for chicken nugget stored at 250 C and 0.17 to 0.43 for the product stored at 40 C across the packaging materials investigated. The water activity and monolayer moisture results of sorption analysis revealed that good packaging materials of very low water vapour transmission rate should be used for storage of chicken nuggets as high-density polyethylene offered a promising alternative than low density polyethylene.

UV Radiation and Protein Hydrolysates in Bio-Based Films: Impacts on Properties and Italian Salami Preservation.

UV radiation was combined with the incorporation of fish protein hydrolysates to improve the performance of active bio-based films for food packaging. UV radiation was not used previously to enhance the packaging performance of blend films of starch/protein, and fish protein hydrolysates were not incorporated in bio-based polymer surfaces previously. Rice starch and fish proteins (from Whitemouth croaker muscle) were utilized to prepare films by the casting technique, which were UV-radiated under different exposure times (1, 5, and 10 min). The packaging performance of the films was determined according to the mechanical and barrier performance, solubility, and color. Fish protein hydrolysates (from Argentine croaker muscle) were then incorporated into the films (bulk structure or surface). The results showed that UV radiation for 1 min increased the tensile strength and modified the optical properties of films. It also altered the structure of the polymeric matrix, as demonstrated by the microstructure and thermal analysis, in agreement with the data obtained in packaging properties. The evaluation of antioxidant capacity through 2,2-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) and reducing power indicated that incorporating fish protein hydrolysates either in the films' bulk structure or film surface promoted antioxidant properties; control films (produced with rice starch/fish proteins without hydrolysates) also presented antioxidant potential. According to the peroxide value and thiobarbituric acid reactive substance (TBARS) assays, control films and the films containing hydrolysates in their bulk structure or on the surface could prevent the lipid oxidation of Italian salami. Thus, combining UV radiation to shape the characteristics of bio-based materials with fish protein hydrolysates to reduce lipid oxidation contributes to the performance of active bio-based films for food packaging.

Identification and inhibition of key off-odorants in boiled greenfin horse-faced filefish (Thamnaconus septentrionalis) subjected to perilla polyphenols extract

The generation of off-flavor during boiling significantly hampers the acceptability and popularity of the greenfin horse-faced filefish (BFF). Adding perilla to Chinese fish dishes can enhance the flavor of fish. A total of 72 volatile compounds were quantified in BFF with different addition of Perilla polyphenol extract (PPE) (i.e., 0, 0.1 %, 0.3 %, 0.5 %, w: w) by headspace solid phase microextraction and gas chromatography-mass spectrometry (HS-SPME-GC-MS). Based on the gas chromatography-olfactometry (GC-O), odor activity value (OAV) and recombination omission testing, (Z)-4-heptenal, (E,E)-2,4-heptadienal, (E,E)-2,4-nonadienal, (E,E)-2,4-decadienal, heptanal, nonanal, octanal, decanal, (E)-2-hexenal, (E)-2-octenal, (E)-2-nonenal were identified as volatiles contributing to the off-flavor, while linalool and β-ionone exhibited a mitigating effect. Moreover, the antioxidant capacities of PPE assessed through 2,2-diphenyl-1-picrylhydrazyl (DPPH) and thiobarbituric acid reactive substances (TBARs) assays, exhibited a positive correlation with perilla polyphenols contents. Orthogonal partial least square discriminate analysis (OPLS-DA) and partial least-squares regression (PLSR) results indicated a significant relationship between the off-flavor of BFF and lipid oxidation, adding 0.3% PPE resulted in a marked improvement in flavor profile, with a reduction in off-flavors associated with fishy, oily/fatty and marine, while simultaneously enhancing the freshness, herbal flavors, and overall acceptability of the product.

SORPTION ISOTHERM AND QUALITY ASSESSMENT OF CHICKEN JERKY

This study investigated the sorption isotherm, proximate composition, lipid oxidation, and sensory evaluation of chicken jerky. The moisture adsorption behavior was analyzed using a gravimetric method while proximate analysis was performed according to official methods. Lipid oxidation was measured by thiobarbituric acid reactive substances (TBARS) assay while sensory evaluation was conducted with trained panelists based on appearance, taste, odor, texture, and overall acceptability. Results showed that equilibrium moisture content increased as water activity increased from 0.043 to 0.750 but remained relatively constant at 0.843 to 0.973 aw. Proximate analysis revealed that the protein content of chicken jerky was high (76.13) while a low-fat content (1.72%) was obtained in this study. Sensory evaluation indicated that overall acceptability was highly dependent on texture followed by aroma and flavour. Based on these results it can be concluded that factors such as formulation, processing method, ingredient quality and raw meat quality greatly influence the physicochemical properties of chicken jerky which ultimately affects consumer acceptance. KEYWORDS: Aroma, Humidity, Salt,Shelf-life, Snack, Storage

Enhancing Raw Bovine Milk Quality using Ultraviolet-C (UV-C) Irradiation: A Microbial and Lipid Peroxidation Study.

This study investigated the efficacy of ultraviolet-C (UV-C) irradiation in enhancing the quality of raw bovine milk by targeting microbial populations and lipid peroxidation, both of which are key factors in milk spoilage. We categorized the raw milk samples into three groups based on initial bacterial load: low (<3 Log 10 CFU/mL), medium (3-4 Log 10 CFU/mL), and high (>4 Log 10 CFU/mL). Using a 144 W thin-film UV-C reactor, we treated the milk with a flow rate of 3 L/min. We measured the bacterial count including standard plate count, coliform count, coagulase-negative staphylococci count, and lactic acid bacteria count and lipid peroxidation (via thiobarbituric acid reactive substances assay) pre- and post-treatment. Our results show that UV-C treatment significantly reduced bacterial counts, with the most notable reductions observed in high and medium initial load samples (>4 and 3-4 Log 10 CFU/mL, respectively). The treatment was particularly effective against coliforms, showing higher reduction efficiency compared to coagulase-negative staphylococci and lactic acid bacteria. Notably, lipid peroxidation in UV-C treated milk was significantly lower than in pasteurized or untreated milk, even after 72 hours. These findings demonstrate the potential of UV-C irradiation as a pre-treatment method for raw milk, offering substantial reduction in microbial content and prevention of lipid peroxidation, thereby enhancing milk quality.

Oxidative stress, microparticles, and E-selectin do not depend on HIV suppression.

Oxidative stress and inflammation are considered predictors of diseases associated with aging. Markers of oxidative stress, inflammation, and endothelial activation were investigated in people with HIV on antiretroviral treatment to determine whether they had an immunosenescent phenotype that might predispose to the development of premature age-related diseases. This study was conducted on 213 subjects with HIV. The control groups consisted of healthy HIV-negative adults. The level of oxidative stress was measured by assessing the production of malondialdehyde levels, which were detected by thiobarbituric acid reactive substance (TBARS) assay. The level of microparticles indicated the presence of inflammation and endothelial activation was measured by E-selectin levels. Significant differences were determined by appropriate statistical tests, depending on the distribution of variables. Relationships between continuous variables were quantified using Spearman's rank correlation coefficient. TBARS, and microparticle and E-selectin levels were significantly higher in untreated and treated subjects with HIV compared with HIV-negative controls (P<0.001). The levels of the investigated markers were not significantly different between untreated and treated patients and no significant correlation of these markers was found with CD4+ count, CD4+/CD8+ ratio, and the number of HIV-1 RNA copies. Elevated markers of oxidative stress, inflammatory and endothelial activation were independent of the virologic and immunologic status of people with HIV. These results support the hypothesis that residual viremia in cellular reservoirs of various tissues is a key factor related to the premature aging of the immune system and predisposition to the premature development of diseases associated with aging.

Fatty Acid Binding Protein-4 Silencing Inhibits Ferroptosis to Alleviate Lipopolysaccharide-induced Injury of Renal Tubular Epithelial Cells by Blocking Janus Kinase 2/Signal Transducer and Activator of Transcription 3 Signaling.

Sepsis-induced kidney injury (SAKI) has been frequently established as a prevailing complication of sepsis which is linked to unfavorable outcomes. Fatty acid-binding protein-4 (FABP4) has been proposed as a possible target for the treatment of SAKI. In the current work, we aimed to explore the role and underlying mechanism of FABP4 in lipopolysaccharide (LPS)-induced human renal tubular epithelial cell damage. In LPS-induced human kidney 2 (HK2) cells, FABP4 expression was tested by the reverse transcription-quantitative polymerase chain reaction and Western blot. Cell counting kit-8 method assayed cell viability. Inflammatory levels were detected using the enzyme-linked immunosorbent assay. Immunofluorescence staining measured the nuclear translocation of nuclear factor kappa B p65. Thiobarbituric acid-reactive substances assay and C11 BODIPY 581/591 probe were used to estimate the level of cellular lipid peroxidation. Fe2+ content was examined by the kit. In addition, the expression of proteins related to inflammation-, ferroptosis- and Janus kinase 2 (JAK2)/signal transducer, and activator of transcription 3 (STAT3) signaling was detected by the Western blot analysis. The results revealed that FABP4 was significantly upregulated in LPS-treated HK2 cells, the knockdown of which elevated the viability, whereas alleviated the inflammation and ferroptosis in HK2 cells challenged with LPS. In addition, down-regulation of FABP4 inactivated JAK2/STAT3 signaling. JAK2/STAT3 stimulator (colivelin) and ferroptosis activator (Erastin) partially restored the effects of FABP4 interference on LPS-triggered inflammation and ferroptosis in HK2 cells. Together, FABP4 knockdown inhibited ferroptosis to alleviate LPS-induced injury of renal tubular epithelial cells through suppressing JAK2/STAT3 signaling.

Ameliorative effect of crocin on post-thaw quality, fertility-associated gene expression and fertilization potential of buffalo (Bubalus bubalis) bull sperm.

Buffalo bull sperm suffer more cryoinjuries due to lipid peroxidation of high structural polyunsaturated fatty acid contents than cattle sperm. Consequently, the post-thaw fertilization potential of buffalo bull sperm is compromised. Crocin is a carotenoid known for its antioxidant potential through scavenging reactive oxygen species. Objectives of the current study were to investigate the effect of crocin addition in the semen extender on post-thaw quality, fertility-associated gene expression and fertilization potential of buffalo bull sperm. Semen samples (n = 32) from four Nili-Ravi buffalo bulls were extended with tris-citric acid extender containing different concentrations of crocin (0 mM; control, 0.5, 1, 1.5 and 2 mM). The extended semen was packed in 0.5 mL French straws (25 × 106 sperm/straw) and cryopreserved in liquid nitrogen. Computer-assisted semen analysis, hypo-osmotic swelling test, normal apical ridge assay, Rhodamine 123, acridine orange, propidium iodide staining, and thiobarbituric acid reactive substances assay were performed to assess sperm motility parameters, plasma membrane integrity, acrosome integrity, mitochondrial membrane potential, DNA integrity, viability, and lipid peroxidation, respectively. Expression levels of sperm acrosome-associated SPACA3, DNA condensation-related PRM1, anti-apoptotic BCL2, pro-apoptotic BAX, and oxidative stress-associated ROMO1 genes were evaluated through qPCR. The fertility of semen doses containing the most potent concentration of crocin (based on optimum post-thaw semen quality) was compared with control during the breeding season. Buffaloes (n = 400; 200/group) were inseminated 24 h after the onset of oestrus and transrectally palpated for pregnancy at least 60 days post-insemination. Results revealed that 0.5 and 1 mM crocin improved sperm post-thaw total motility, plasma membrane integrity, acrosome integrity, mitochondrial membrane potential and viability, and 1 and 1.5 mM crocin enhanced catalase activity and reduced lipid peroxidation compared to control (p < .05). Moreover, 1 mM crocin improved sperm post-thaw progressive motility, kinematics, and DNA integrity, and 1.5 mM crocin enhanced plasma membrane integrity than control (p < .05). Expression levels of SPACA3, PRM1 and BCL2 genes were higher (p < .05) with 1 mM crocin compared to other groups. In contrast, no difference (p > .05) was noticed in expressions of BAX and ROMO1 genes among all groups. The fertility rate of semen doses containing the most potent concentration (1 mM) of crocin was higher (p = .0465) compared to control (56 ± 0.03% vs. 46 ± 0.04%, respectively). In conclusion, 1 mM crocin in the semen extender improves post-thaw quality, fertility-associated gene expression and fertilization potential of buffalo bull sperm.

Sestrin2 attenuates depressive-like behaviors and neuroinflammation in CUMS mice through inhibiting ferroptosis.

Sestrin2 (SESN2) is a stress-inducible protein and acts as a neuroprotective regulator. The present study aimed to explore the antidepressant activity of SESN2 and its relevant mechanism. Depression mouse model was established by chronic unpredictable mild stress (CUMS) for a successive 5 weeks. Behaviors tests were conducted to examine depressive-like behaviors including sugar preference test, tail suspension test and open field test. The expression of SESN2 and ferroptosis-related proteins was examined by western blot. The production of cytokines was measured by ELISA. Iron deposition was assessed using Prussian blue staining and Fe 2+ content was measured using commercial kits. Lipid peroxidation was evaluated by thiobarbituric acid reactive substances assay. BV-2 cells were treated with LPS to induce microglial activation, which was evaluated by the iba-1 level adopting immunofluorescence assay. The ferroptosis inducer Erastin was adopted for the pretreatment in BV-2 cells to conduct a rescue experiment. SESN2 was downregulated in CUMS-induced mice, and SESN2 overexpression dramatically ameliorated CUMS-induced depression-like behaviors. Meanwhile, SESN2 reduced the production of pro-inflammatory cytokines and iba-1 level in hippocampus of CUMS mice, as well as reducing iron deposition and lipid peroxidation, demonstrating that SESN2 reduced microglial activation, neuroinflammation and ferroptosis in CUMS mice. Similarly, SESN2 also restricted iba-1 level, pro-inflammatory cytokines production, and ferroptosis in LPS-induced BV-2 cells, which was partly reversed by additional treatment of Erastin. These findings suggest that SESN2 possesses potent antidepressant property through inhibiting ferroptosis and neuroinflammation.

PSVIII-19 Saccharomyces Cerevisiae Fermentation Product Modulates Blood Cell Counts and Gut Immunity in Healthy Adult Cats

Abstract Dietary Saccharomyces cerevisiae fermentation product (SCFP) has been shown to modulate immune responses in dogs and livestock. Information about feeding SCFP to cats, however, is lacking. This study aimed to evaluate the effects of SCFP (TruMune, Cargill, Inc.) on immune indices including fecal IgA, serum oxidative stress markers, and toll-like receptor (TLR) responsiveness in healthy adult cats. Sixty-three cats (mean age = 6.3 ± 2.8 yr; mean BW = 4.4 ± 0.1 kg) were used in a randomized complete block design. All cats were fed a control kibble diet without SCFP (CON) during a 21-day adaptation period. Cats were randomly assigned to one of the three diets including CON, a dry diet containing 150 mg/kg BW SCFP (TRUL), and a dry diet containing 300 mg/kg BW SCFP (TRUH) and fed for 42 days. Fecal samples were collected on days 0, 21, and 42 for fecal IgA analysis. Serum samples were collected on days 0 and 42 for measurements of serum chemistry, complete blood count, and oxidative stress markers. Whole blood samples were collected on day 42 for TLR responsive evaluation. Protocols were approved by the facility’s Institutional Animal Care and Use committee before the study. Data were analyzed using R v.4.1.2. Differences P &amp;lt; 0.05 were considered significant and P &amp;lt; 0.10 as trends. All serum chemistry and blood cell count values were within the reference ranges, with serum chemistry not affected by SCFP. Absolute cell counts of eosinophils and neutrophils decreased linearly with SCFP dose (P &amp;lt; 0.001) while absolute cell counts of lymphocytes and monocytes increased linearly with SCFP dose (P &amp;lt; 0.01). TLR responsiveness was not altered (P &amp;gt; 0.05) by SCFP. A tendency (P = 0.08) of quadratic treatment dose response was noted for thiobarbituric acids from the thiobarbituric acid reactive substance assay, with cats fed TRUL having the greatest thiobarbituric acid values. Fecal IgA showed a quadratic treatment dose response on day 42 (P = 0.01), with cats fed TRUL having the lowest fecal IgA values. These results demonstrate that SCFP may affect immune outcomes including blood cell counts and fecal IgA in adult healthy cats.

Corylus avellana "Nocciola Piemonte": metabolomics focused on polar lipids and phenolic compounds in fresh and roasted hazelnuts.

The common hazel plant (Corylus avellana L., Betulaceae) is one of the most popular tree nuts widespread in Europe and Asia. In Italy, there are different cultivars among which the cultivar affording the valuable hazelnut "Tonda Gentile Trilobata," also known as "Tonda Gentile delle Langhe," covered by the Protected Geographical Indication (PGI) label "Nocciola Piemonte" (NP), known for its sweetness, cooked-bread aroma, and the low intensity of the burnt aroma. In order to obtain a detailed and in-depth characterization of the polar fraction of fresh (NPF) and roasted (NPR) kernels of NP the analysis of the n-butanol extracts by liquid chromatography coupled to electrospray ionization and high-resolution mass spectrometry (LC-ESI/HRMS) was carried out. Moreover, to evaluate the quantitative distribution of the most representative polar lipids in NPF and NPR, the analysis by liquid chromatography combined with tandem mass spectrometry (LC-MS/MS) was performed. To unambiguously identify the phenolic compounds highlighted by the LC-ESI/HRMS profiles, they were isolated from the n-butanol extract and characterized by Nuclear Magnetic Resonance (NMR) experiments. Finally, the ability of the isolated compounds to exert radical scavenging activity and to inhibit the lipid peroxidation induced by H2O2 or H2O2/Fe2+ was tested by Trolox Equivalent Antioxidant Capacity (TEAC) and thiobarbituric acid reactive substances (TBARS) assays, respectively. The LC-ESI/HRMS allowed to ascertain the presence of phenolic compounds and multiple classes of polar lipids including phospholipids, glycolipids, sphingolipids, and oxylipins. The quantitative analysis highlighted in NPR fraction a lipid content three times higher than in NPF, evidencing lyso-phospholipids and phospholipids as the most represented lipid classes in both NPF and NPR, together accounting for 94 and 97% of the considered lipids, respectively. Furthermore, phytochemical analysis permitted to identify flavonoid and diarylheptanoid derivatives. In particular, quercetin 3-O-β-D-galactopyranosyl-(1→2)-β-D-glucopyranoside and myricetin-3-O-α-L-rhamnopyranoside showed the highest antioxidant activity, exhibiting TEAC values similar to that of quercetin, used as reference compound (2.00 ± 0.03 and 2.06 ± 0.03 mM vs 2.03 ± 0.03 mM, respectively). Moreover, most of the tested compounds were found to reduce lipid peroxidation induced by H2O2 and H2O2/Fe2+ more than curcumin used as positive control, with myricetin-3-O-α-L-rhamnopyranoside determining 44.4 % and 34.1 % inhibition percentage, respectively.

Downwind gas condensate volatiles affect phytoplankton communities

We investigated the effects of volatile organic carbons (VOCs) evaporated from gas condensate on the cyanobacteria Synechococcus sp. WH8103, the diatom Asterionellopsis glacialis, and the dinoflagellate Alexandrium minutum. We used custom algal incubation chambers enabling only the gas condensate-derived VOCs to interact with the cell cultures via an atmospheric bridge, without direct contact with the hydrocarbon oil. The exposure to gas condensate VOCs reduced the abundance, growth rate, and photosynthetic efficiency of Synechococcus sp. WH8103. Thiobarbituric acid reactive substances (TBARS) assays hint at oxidative damage to the chloroplasts and/or the thylakoid membranes in this organism. A.glacialis abundance, physiological state and growth rates remained unchanged, whereas A.minutum abundance and photosynthetic efficiency increased relative to their respective controls. Our results demonstrate that the effects of a gas condensate formed due to an oil spill will not be restricted to the polluted area, but may be prominent in downwind locations through atmospheric transport.

Investigating physiological responses of Wild Rocket subjected to artificial Ultraviolet B irradiation

Utilization of ultraviolet-B radiation (UV-B) is a way to encourage the accumulation of phytochemicals such as glucosinolates, carotenoids, phenolics and flavonoids in plants. In the current research, UV-B treatments were performed, as a possible elicitation strategy to promote the secondary metabolites accumulation in wild rocket (Diplotaxis tenuifolia (L.) DC). After performing the destructive analysis, it has been found that the concentration of chlorophyl a and b as well as carotenoids decreased in response to the UV-B stress. Thiobarbituric acid reactive substances (TBARS) assay has been performed to investigate the membranes damage caused by the UV-B radiation. The analyses confirmed that the highest UV-B tested dose induced the greater damage. Interestingly, an increase of phenolic index, and an accumulation of anthocyanins and glucosinolates has been recorded under the successive exposure of varying UV-B doses. The total sugars, reducing sugars and sucrose buildup were also remarkable. Non-destructive analysis revealed a decline in the overall performance of the UV-B treated plants as well as the maximum quantum efficiency of photosystem II. A decrease in the chlorophyll estimates while an enhanced flavanols and anthocyanins were observed through non-destructive readings. Results of this study not only provided a feasible UV-B dose selection and supplementation for the wild rocket but also the understanding about enhanced phytochemical accumulation in plants against UV-B stress.

Antioxidative Properties of Fermented Soymilk Using Lactiplantibacillus plantarum LP95.

In recent times, there has been a growing consumer interest in replacing animal foods with alternative plant-based products. Starting from this assumption, for its functional properties, soymilk fermented with lactic acid bacteria is gaining an important position in the food industry. In the present study, soymilk was fermented with Lactiplantibacillus plantarum LP95 at 37 °C, without the use of stabilizers as well as thickeners and acidity regulators. We evaluated the antioxidant capacity of fermented soymilk along with its enrichment in aglycone isoflavones. The conversion of isoflavone glucosides to aglycones (genistein, glycitein, and daidzein) was analyzed together with antioxidant activity (ABTS) measurements, lipid peroxidation measurements obtained by a thiobarbituric acid reactive substance (TBARS) assay, and apparent viscosity measurements. From these investigations, soymilk fermentation using Lp. plantarum LP95 as a starter significantly increased isoflavones' transformation to their aglycone forms. The content of daidzein, glycitein, and genistein increased after 24 h of fermentation, reaching levels of 48.45 ± 1.30, 5.10 ± 0.16, and 56.35 ± 1.02 μmol/100 g of dry weight, respectively. Furthermore, the antioxidant activity increased after 6 h with a reduction in MDA (malondialdehyde). The apparent viscosity was found to increase after 24 h of fermentation, while it slightly decreased, starting from 21 days of storage. Based on this evidence, Lp. plantarum LP95 appears to be a promising candidate as a starter for fermented soymilk production.

Ginsenoside Rh4 inhibits the malignant progression of multiple myeloma and induces ferroptosis by regulating SIRT2.

Multiple myeloma (MM) has a high mortality rate, and the exploration of therapeutic drugs for MM with low side effects is a hot topic at the moment. Ginsenoside Rh4 has been shown to inhibit tumour growth in many cancers. However, the role of ginsenoside Rh4 in MM and its reaction mechanism have not been reported so far. After the treatment with different concentrations of ginsenoside Rh4, the proliferation of NCI-H929 cells was detected by Cell Counting Kit-8 and 5-ethynyl-2'-deoxyuridine staining. The cell apoptosis and cycle arrest were detected by flow cytometry and western blot. The thiobarbituric acid-reactive substances (TBARS) production was assessed with TBARS assay, whereas Fe2+ fluorescence assay was used for the measurement of Fe2+ level. The production of reactive oxygen species was evaluated with dichloro-dihydro-fluorescein diacetate staining, and western blot was applied for the estimation of ferroptosis-related proteins. The potential targets of ginsenoside Rh4 were predicted by molecular docking technology and verified by western blot. The transfection efficacy of overexpression-SIRT2 was examined with quantitative reverse transcription polymerase chain reaction and western blot. To figure out the detailed reaction mechanism between ginsenoside Rh4 and SIRT2 in MM, rescue experiments were conducted. We found that ginsenoside Rh4 inhibited cell proliferation, induced cell apoptosis, promoted cycle arrest and facilitated ferroptosis in MM. Moreover, ginsenoside Rh4 inhibited SIRT2 expression in MM cells. The overexpression of SIRT2 reversed the effects of ginsenoside Rh4 on cell proliferation, cell apoptosis, cycle arrest and ferroptosis in MM. Overall, ginsenoside Rh4 inhibited the malignant progression of MM and induced ferroptosis by regulating SIRT2.

The cytotoxicity effect of 7-hydroxy-3,4-dihydrocadalene from Heterotheca inuloides and semisynthetic cadalenes derivates towards breast cancer cells: involvement of oxidative stress-mediated apoptosis.

Heterotheca inuloides, traditionally employed in Mexico, has demonstrated anticancer activities. Although it has been proven that the cytotoxic effect is attributed to cadinane-type sesquiterpenes such as 7-hydroxy-3,4-dihydrocadalene, the mechanism of action by which these agents act in tumor lines and their regulation remain unknown. This study was undertaken to investigate for first time the cytotoxic activity and mechanism of action of 7-hydroxy-3,4-dihydrocadalene and two semi-synthetic cadinanes derivatives towards breast cancer cells. Cell viability and proliferation were assayed by thiazolyl blue tetrazolium bromide (MTT) assay and Trypan blue dye exclusion assay. Cell migration measure was tested by wound-healing assay. Moreover, the reactive oxygen species (ROS) and lipid peroxidation generation were measured by 2',7'-dichlorofluorescein diacetate (DCFH-DA) assay and thiobarbituric acid reactive substance (TBARS) assay, respectively. Furthermore, expression of caspase-3, Bcl-2 and GAPDH were analyzed by western blot. The results showed that 7-hydroxy-3,4-dihydrocadalene inhibited MCF7 cell viability in a concentration and time dependent manner. The cytotoxic potency of semisynthetic derivatives 7-(phenylcarbamate)-3,4-dihydrocadalene and 7-(phenylcarbamate)-cadalene was remarkably lower. Moreover, in silico studies showed that 7-hydroxy-3,4-dihydrocadalene, and not so the semi-synthetic derivatives, has optimal physical-chemical properties to lead a promising cytotoxic agent. Further examination on the action mechanism of 7-hydroxy-3,4-dihydrocadalene suggested that this natural product exerted cytotoxicity via oxidative stress as evidenced in a significantly increase of intracellular ROS levels and in an induction of lipid peroxidation. Furthermore, the compound increased caspase-3 and caspase-9 activities and slightly inhibited Bcl-2 levels. Interestingly, it also reduced mitochondrial ATP synthesis and induced mitochondrial uncoupling. Taken together, 7-hydroxy-3,4-dihydrocadalene is a promising cytotoxic compound against breast cancer via oxidative stress-induction.

Supramolecular complexes between Plinia cauliflora (DC.) Kausel extracts and β-cyclodextrin: Physicochemical characterization and antioxidant and anti-inflammatory properties

This work aims to investigate the supramolecular complexes formation between β-cyclodextrin (βCD) and compounds present in leaves (EEL) and branches (EEB) of Plinia cauliflora (DC.) Kausel Physicochemical characterization was carried out to investigate the formation of supramolecular complexes (EEL/βCD and EEB/βCD), such as, Fourier-transform infrared (FTIR) spectroscopy, thermal analysis (TGA/DTA), scanning electron microscopy (SEM), dynamic light scattering (DLS), Zeta potential (ZP), and electrical conductivity measurements. Antioxidant activity was evaluated by the 2,2-diphenyl-1-picrylhydrazyl, β-carotene/linoleic acid, and thiobarbituric acid reactive substance assays. Finally, anti-inflammatory activity was investigated by nitric oxide (NO) determination and evaluation of ear edema inhibition. FTIR spectroscopy and TGA/DTA confirmed the interactions in the solid-state, while ZP and DLS showed that the interaction between extracts and βCD caused differences in the hydrophobic nanoprecipitates. Moreover, the SEM micrograph of EEL/βCD and EEB/βCD indicates changing in particle morphology which confirms the interactions between extract compounds and βCD. EEL/βCD and EEB/βCD showed promising antioxidant activity for all assays performed. Furthermore, anti-inflammatory activity was confirmed by a significant reduction of NO (maximum of 90 and 85% for EEL/βCD and EEB/βCD, respectively) and inhibition of ear edema above 93% for both phytocomplexes. The development of a formulation based on the molecular encapsulation of extracts from P. cauliflora with βCD could be an effective, safe, and sustainable therapeutic alternative. Therefore, our samples showed the ability to reduce oxidative stress and inflammation, and serve as potential anti-inflammatory agents.

8-Hydroxy-2'-deoxyguanosine and malondialdehyde in plasma and their association with disease severity in 20 cats with chronic kidney disease.

Oxidative stress is associated with the development and progression of chronic kidney disease (CKD) in humans. The aim of this study was to evaluate the concentrations of oxidative stress markers, 8-hydroxy-2'-deoxyguanosine (8-OHdG) and malondialdehyde (MDA), in the plasma and urine of cats with different stages of CKD. Plasma and urine samples were collected from cats with CKD that were referred to the Veterinary Medical Center of the University of Tokyo between April 2019 and October 2022. Plasma and urine samples were collected from healthy cats (n = 6 at most), cats with stage 2 CKD (n = 8) and stage 3-4 CKD (n = 12), and cats with idiopathic cystitis (disease control, n = 5). Plasma and urine concentrations of 8-OHdG and MDA were measured using ELISA and thiobarbituric acid reactive substances assay kits, respectively. The median plasma 8-OHdG concentrations were 0.156 ng/ml (<0.125-0.210 ng/ml) in the healthy group, <0.125 ng/ml (range <0.125 ng/ml) in the idiopathic cystitis group, 0.246 ng/ml (range 0.170-0.403 ng/ml) in cats with stage 2 CKD and 0.433 ng/ml (range 0.209-1.052 ng/ml) in cats with stage 3-4 CKD. Concentrations in stage 3-4 CKD were significantly higher than those in the healthy and disease control groups. Plasma MDA concentrations were low in the healthy and disease control groups and significantly higher in cats with stage 3-4 CKD. In every cat with CKD, plasma 8-OHdG and MDA concentrations were positively correlated with plasma creatinine concentrations (8-OHdG, rs = 0.68; MDA, rs = 0.67). Urinary 8-OHdG/urinary creatinine (u-CRE) and urinary MDA/u-CRE levels did not differ significantly between the groups; however, it was difficult to evaluate them because of the small sample size. This report shows that plasma 8-OHdG and MDA concentrations increase with the severity of feline CKD. These markers may be useful for assessing oxidative stress in cats with CKD.

Inhibitory effect of Solanum anguivi fruit extracts against lead-induced lipid peroxidation in rat liver and kidney

Rapid incidence of liver and kidney disorders resulting in high mortality nowadays is of great concern and there is this very need to salvage the sudden death resulting from the disorders. Lipid peroxidation which is the hallmark of oxidative stress that damage body tissues is prominent as one of the causative factors leading to many chronic and degenerative diseases. This study is sought to investigate the inhibitory effect of aqueous and ethanolic extracts of Solanum Anguivi fruit against lead acetate induced lipid peroxidation in the liver and kidney tissues of albino rats in vitro using thiobarbituric acid reactive substances (TBARS) assay. The results obtained indicate a concentration dependent increase in inhibitory potential of the extracts against lead acetate in both liver and kidney tissues. Highest extract inhibitory potential was (90%) at highest extract concentration (160mg/ml) and this was obtained in the liver tissue. Hence, the results from this study suggest that oxidative stress in liver and kidney tissues could be potentially prevented or managed by dietary intake of Solanum Anguivi fruits.

Carbon Nanodots Attenuate Lipid Peroxidation in the LDL Receptor Knockout Mouse Brain.

Abnormal cholesterol metabolism can lead to oxidative stress in the brain. Low-density lipoprotein receptor (LDLr) knockout mice are models for studying altered cholesterol metabolism and oxidative stress onset in the brain. Carbon nanodots are a new class of carbon nanomaterials that possess antioxidant properties. The goal of our study was to evaluate the effectiveness of carbon nanodots in preventing brain lipid peroxidation. LDLr knockout mice and wild-type C57BL/6J mice were treated with saline or 2.5 mg/kg bw of carbon nanodots for a 16-week period. Brains were removed and dissected into the cortex, midbrain, and striatum. We measured lipid peroxidation in the mouse brain tissues using the Thiobarbituric Acid Reactive Substances Assay and iron and copper concentrations using Graphite Furnace Atomic Absorption Spectroscopy. We focused on iron and copper due to their association with oxidative stress. Iron concentrations were significantly elevated in the midbrain and striatum of the LDLr knockout mice compared to the C57BL/6J mice, whereas lipid peroxidation was greatest in the midbrain and cortex of the LDLr knockout mice. Treatment with carbon nanodots in the LDLr knockout mice attenuated both the rise in iron and lipid peroxidation, but they had no negative effect in the C57BL/6J mice, indicating the anti-oxidative stress properties of carbon nanodots. We also assessed locomotor and anxiety-like behaviors as functional indicators of lipid peroxidation and found that treatment with carbon nanodots prevented the anxiety-like behaviors displayed by the LDLr knockout mice. Overall, our results show that carbon nanodots are safe and may be an effective nanomaterial for combating the harmful effects caused by lipid peroxidation.