B. Isolation, purification, characterization, and assay of antioxygenic enzymes[13] Catalase in vitro
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Catalase exerts a dual function: (1) decomposition of H2O2 to give H2O and O2 (catalytic activity) and (2) oxidation of H donors, for example, methanol, ethanol, formic acid, phenols, with the consumption of 1 mol of peroxide (peroxide activity). The kinetics of catalase does not obey the normal pattern. Measurements of enzyme activity at substrate saturation or determination of the Ks is therefore impossible. In contrast to reactions proceeding at substrate saturation, the enzymic decomposition of H2O2 is a first-order reaction, the rate of which is always proportional to the peroxide concentration present. Consequently, to avoid a rapid decrease in the initial rate of the reaction, the assay must be carried out with relatively low concentrations of H2O2 (about 0.01 M). This chapter discusses the catalytic activity of catalase. The method of choice for biological material, however, is ultraviolet (UV) spectrophotometry. Titrimetric methods are suitable for comparative studies. For large series of measurements, there are either simple screening tests, which give a quick indication of the approximative catalase activity, or automated methods.
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A total of 84 active ingredients and 830 target genes were screened in BYHWD, among which 56 target genes intersected with DPF-related targets. Network pharmacological analysis revealed that the active ingredients can regulate target genes such as IL-6, TNF-α, VEGFA and CASP3, mainly through AGE-RAGE signaling pathway, HIF-1 signaling pathway and TNF signaling pathway. Molecular docking and molecular dynamics simulations suggested that IL6-astragaloside IV, IL6-baicalein, TNFα-astragaloside IV, and TNFα-baicalein docking complexes could bind stably. Animal experiments showed that BYHWD could reduce the expression of core targets such as VEGFA, CASP3, IL-6 and TNF-α. In addition, BYHWD could reduce blood glucose, lipid, and MDA levels in DPF while increasing the activities of SOD, CAT and GSH-Px. BYHWD attenuated the expression of HYP and collagen I, mitigating pathological damage and collagen deposition within lung tissue.
BYHWD modulates lipid metabolism disorders and oxidative stress by targeting the core targets of IL6, TNF-α, VEGFA and CASP3 through the AGE-RAGE signaling pathway, making it a potential therapy for DPF.
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2024, RhizosphereThis study investigated the efficacy of 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase-producing rhizobacterial strains isolated from soybean rhizosphere to alleviate the negative impact of drought stress in soybean. Twenty rhizobacterial strains were isolated from disease-resistant soybeans, and only four isolates displayed ACC deaminase activity (more than 500 nmol of ketobutyrate mg protein−1 h−1). These strains also exhibited a variety of plant growth-promoting (PGP) traits, such as ammonia production, indole acetic acid concentration (>35 g mL−1), hydrogen cyanide (HCN) production, and solubilization of inorganic calcium orthophosphate (>70 μg mL−1). Based on 16S rDNA gene sequencing, the bacterial isolates (AKAD 1-2, AKAD 1-3, AKAD 3-1, and AKAD 3-7) were identified as Pantoea agglomerans (MH304295), Bacillus subtilis (MH304311), Bacillus cereus (MH333217), and Bacillus licheniformis (MH304284), respectively. .Seed bio-priming with these rhizobacteria improvedin vivodrought tolerance, mitigated drought stress and increased shoot length (16–68.6%), root length (40–108%), proline content (14.87–42.69%), sugar content (11.36–67.3%), and protein content (11.77–45%). The potential enzymatic activities of catalase (CAT), superoxide dismutase (SOD), and peroxidase (POD) also increased by 35.87%, 26.63%, and 15.15%, respectively, in drought-stressed plants. The results support the efficacy of these isolates as effective bio-stimulants for improving crop performance in drought-affected agricultural fields. Thus, the present work opens new avenues for harnessing the potential of rhizobacterial bio-stimulants to address the challenges posed by drought stress in soybean cultivation, thereby advancing sustainable agricultural strategies for future food security.
Thermal tolerance of a freshwater amphipod Gammarus lacustris can be enhanced by acclimation to higher mineralization
2024, Comparative Biochemistry and Physiology -Part A : Molecular and Integrative PhysiologyTemperature and mineralization are among the most important environmental factors affecting all processes of aquatic ecosystems, including geographical distribution of water animals. Previously we showed that a brackish water population of Gammarus lacustris, a widespread amphipod, demonstrates substantially higher thermotolerance than a freshwater population. A possible reason for this difference is the fact that brackish water conditions are closer to internal media mineralization. Here we aimed to test this hypothesis and relate the observed effects in animal survival under the heat shock to the status of cellular defence systems. We acclimated four groups of amphipods from the same freshwater population to 0.5 ‰ and 15 ‰ at the temperatures of 6°С or 15°С. Acclimation at 6°С, but not at 15°С, to 15 ‰ significantly increased resistance of the amphipods to heat shock at 30°C. At 6°С activities of antioxidant enzymes and levels of the lipid peroxidation products in G. lacustris did not react to the increase in mineralization and the heat shock, while the level of HSP70 elevated two-fold in amphipods acclimated to mineralization of 15 ‰ compared to animals acclimated to 0.5 ‰. Thus, the observed increase in thermotolerance could be explained by the higher initial level of HSP70 and potentially other heat shock proteins caused by a less energy-demanding, more isotonic level.
Diet with different concentrations of lychee peel flour modulates oxidative stress parameters and antioxidant activity in zebrafish
2024, Comparative Biochemistry and Physiology Part - B: Biochemistry and Molecular BiologyThe agri-food industry generates substantial waste, leading to significant environmental impacts. Lychee (Litchi chinensis Sonnerat), which is rich in bioactive compounds in its peel, pulp, and seeds, offers an opportunity for waste use. This study aimed to evaluate the effects of supplementing a high-carbohydrate diet with varying levels of lychee peel flour on lipid metabolism biomarkers and oxidative stress in a zebrafish (Danio rerio) model. A total of 225 zebrafish, approximately four months old, were divided into five groups: control, high-carbohydrate (HC), HC2%, HC4%, and HC6%. The study did not find significant differences in the growth performance of zebrafish in any group. However, the HC6% group exhibited a significant decrease in glucose and triglyceride levels compared with the HC group. Furthermore, this group showed enhanced activities of the antioxidant enzymes catalase (CAT) and superoxide dismutase (SOD), along with reduced levels of malondialdehyde (MDA). Increased antioxidant activity was also evidenced by DPPH−, ABTS+, and β-carotene/Linoleic acid assays in the HC6% group. A positive correlation was identified between SOD/CAT activity and in vitro antioxidant assays. These findings suggest that dietary supplementation with 6% lychee peel flour can significantly modulate glucose homeostasis, lipid metabolism, and antioxidant activity in zebrafish.
Neuroprotective aftermath of Monodora myristica and Glycyrrhiza glabra against cassava diet containing vacuum gas oil induced brain injury in Wistar rats
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