Changes In Enzyme Activities Research Articles

Structural, defense enzyme activity and chemical composition changes in the xylem of Aquilaria sinensis during fungus induction

Agarwood is a dark resinous wood, of high quality and in a short period, can be formed after inoculation with the efficient fungus in Aquilaria sinensis. However, the action site and mechanism of the fungus inducing the agarwood formation remain unclear. In this work, the efficient fungal infection was used to induce agarwood formation on A. sinensis, and samples were collected according to inoculation time. Subsequently, the xylem structure, histochemistry, defense enzyme activities, and the accumulation and in-situ distribution of chemical composition were investigated. The transition layer (TL) was the critical layer for the continuous accumulation of agarwood, and began to appear in the injection of fungal inducers for 15 days (15D). The parenchyma cells of the xylem rays and interxylary phloem were the main action site of fungus in the formation and accumulation of agarwood. The sesquiterpenes and 2-(2-phenylethyl) chromones (PECs) were mainly accumulated in the agarwood layer (AL) and decay layer (DL), while their relative contents were highest in the injection of fungal inducers for 30 days and 180 days (30D and 180D), respectively. Among them, PECs appeared earlier than sesquiterpenes, and both were flindersia-type PECs. They were mainly concentrated in the cell corners and intercellular layer of the cell wall. In addition, Redundancy analysis (RDA) and Mantel test revealed that the different layer percentage (DLP), phenylalanine ammonia lyase (PAL), and starch grains (SG) were the drivers of PECs, and catalase (CAT) was the driver of sesquiterpenes. Therefore, this study will provide a theoretical foundation for revealing the mechanism of fungus-induced agarwood formation and inducing high-quality and high-yielding agarwood in the future.

Environmental pollutant sodium omadine: toxic effects in zebra fish (Danio rerio)

In recent years one of the most striking results of over-population and consumption activities in the world is the rapid increase in environmental pollutants. Environmental pollutants, one of the harmful consequences of technological and modern life, threaten the health of people and other living organisms. In this study, we aimed to determine the effects of sodium omadine (NaOM) on superoxide dismutase enzyme (SOD) activity as an antioxidant and on 8-OHdG levels as oxidative DNA damage in zebrafish. Zebrafish, obtained from the aquarium fish producer, were stocked in experimental aquariums to ensure their adaptation period to the experimental conditions 15 days before the experiment. The fish were exposed to 1 ug/L and 5 ug/L concentrations of NaOM for 24, 72, and 96 h. SOD enzyme activity (U/100 mg tissue) and 8-OHdG (pg/100 mg tissue) were measured using commercial kits. The statistically significant differences in tissue SOD levels and data for DNA damage between the groups were determined as time and dose-dependent (p < 0.05). Biocidal products are environmental pollutants that cause changes in antioxidant enzyme activities, especially in non-target organisms. Marine pollution and the degradation of ecosystems directly affect people, and the results of the study offer awareness of health problems, environmental pollution, and marine pollution.

Screening and Regulatory Mechanisms of Inter-Root Soil Nematicidal Bacteria of Pinus massoniana

Pine Wilt Disease (PWD), caused by the pathogenic nematode Bursaphelenchus xylophilus, is a systemic infectious disease commonly referred to as the “cancer” of pine trees. This devastating disease has gained this analogy due to its ability to rapidly spread within pine populations, leading to substantial losses in forest resources. The primary objective of this study is to investigate the bioprotective potential and underlying mechanisms of action exhibited by rhizosphere microorganisms associated with Masson pine (Pinus massoniana) in the context of controlling the pine wilt nematode. In this experiment, using high-throughput sequencing, significant differences were observed in the rhizosphere soil microbial communities among healthy Masson pine, standing dead trees, and diseased Masson pine. Furthermore, it was found that these microbial communities exhibited distinct community structures at different levels. This study successfully isolated and screened three strains of highly effective nematophagous bacteria from the rhizosphere soil. The identified strains were Lysinibacillus capsici, Bacillus Paramycoides, and Delftia tsuruhatensis. After applying the bacterial suspensions and fermentation extracts of these three strains to the roots of two-year-old Masson pine seedlings, followed by inoculation with pine wilt nematodes after a four-day period, distinct defense responses were observed in the Masson pine. Notably, the activities of phenylalanine ammonia-lyase (PAL) and peroxidase (POD) were significantly increased, leading to a substantial reduction in the incidence of pine wilt disease. Based on the changes in defense enzyme activities, it can be concluded that the fermentation extract of the Lysinibacillus capsici strain exhibits effective nematocidal effects and induces resistance. The significant biological control efficacy and induction of host defense activity indicate the potential application value of this strain and its metabolites as a biocontrol agent for pine wilt disease.

Mechanisms of Digestive Enzyme Response to Acute Salinity Stress in Juvenile Yellowfin Tuna (Thunnus albacares).

This study investigates the effect of a sudden change in salinity for 48 h on the digestive enzyme activity of juvenile yellowfin tuna. The treatment included a control salinity of 32‱ in natural seawater and an experimental salinity of 29‱. Acute stress experiments were carried out on 72 juvenile yellowfin tuna (646.52 ± 66.32 g) for 48 h to determine changes in digestive enzyme activity in different intestinal sections over time (0 h, 12 h, 24 h, 48 h). The activities of pepsin, trypsin, α-amylase, lipase, and chymotrypsin in the digestive organs (stomach, foregut, and pyloric ceca) of juvenile yellowfin tuna were measured. Pepsin and pancreatic protease in the experimental group were significantly lower than in the control group (p < 0.05). α-amylase showed a fluctuating trend of decreasing and then increasing, and its activity trend was pyloric ceca > foregut > stomach. The lipase activity of gastric tissues decreased at the beginning and then increased, reaching a minimum at 24 h (2.74 ± 1.99 U·g protein-1). The change of lipase in the pyloric ceca and foregut was increasing and then decreasing. The lipase activity trend was pyloric ceca > foregut > stomach. The chymotrypsin showed a decreasing and increasing trend and then stabilized at 48 h with a pattern of pyloric ceca > foregut > stomach. Similarly, the gut villi morphology was not significantly altered in the acutely salinity-stressed compared to the non-salinity-stressed. This study suggests that salinity may change the digestive function of juvenile yellowfin tuna, thereby affecting fish feeding, growth, and development. On the contrary, yellowfin tuna is highly adapted to 29‱ salinity. However, excessive stress may negatively affect digestive enzyme activity and reduce fish digestibility. This study may provide a scientific basis for a coastal aquaculture water environment for yellowfin tuna farming, which may guide the development and cultivation of aquaculture.

Flexible changes to the Heliothis virescens ascovirus 3h (HvAV-3h) virion components affect pathogenicity against different host larvae species

ABSTRACT The pathogenicity of a virus to a specific host species is an inerratic and describable ability of a virus to cause infection but is generally shaped by a variety of abiotic and biotic factors. In this investigation, the variations in pathogenicity of Heliothis virescens ascovirus 3h (HvAV-3h) to five noctuid pests were assessed based on mass spectrometry analysis on the virion compositions. Twenty-nine common HvAV-3h proteins were shared across all hosts, and different flexible proteins were identified in the virions of each specific host. Different host proteins were identified as HvAV-3h virion-associated proteins, including different detoxification enzyme proteins. Furthermore, a relatively fixed relationship between viral replication and changes in host detoxification enzyme activity caused by deficiencies in various viral structural proteins was found in the host larvae using a correlation matrix analysis: the host larval carboxylesterase and cytochrome P450 monooxygenases generally had highly similar responses to the viruses blocked by different structural proteins’ antisera and their effects on viral DNA replication. Different interaction patterns for the virion structural proteins were found in different host larvae-produced virions, and the interactions between Spodoptera litura glutathione S-transferases and viral structural proteins were confirmed. The different host responses after viral infection could be the reason for the changes in viral pathogenicity, while the virus responses gradually adapted to the different hosts and there were flexible changes in the virion structures. IMPORTANCE Different pathogenic processes of a virus in different hosts are related to the host individual differences, which makes the virus undergoes different survival pressures. Here, we found that the virions of an insect virus, Heliothis virescens ascovirus 3h (HvAV-3h), had different protein composition when they were purified from different host larval species. These “adaptive changes” of the virions were analyzed in detail in this study, which mainly included the differences of the protein composition of virions and the differences in affinity between virions and different host proteins. The results of this study revealed the flexible changes of viruses to help themselves adapt to different hosts. Also, these interesting findings can provide new insights to improve our understanding of virus adaptability and virulence differentiation caused by the adaptation process.

Cell-Free Supernatant of Lactiplantibacillus plantarum 90: A Clean Label Strategy to Improve the Shelf Life of Ground Beef Gel and Its Bacteriostatic Mechanism.

Lactic acid bacteria metabolites can be used as a clean-label strategy for meat products due to their "natural" and antibacterial properties. In this study, the feasibility of using cell-free supernatant of Lactiplantibacillus plantarum 90 (LCFS) as a natural antibacterial agent in ground beef was investigated. The sensitivity of LCFS to pH, heat and protease, as well as the changes of enzyme activities of alkaline phosphatase (AKP) and Na+/K+-ATP together with the morphology of indicator bacteria after LCFS treatment, were analyzed to further explore the antibacterial mechanism of LCFS. The results showed that the addition of 0.5% LCFS inhibited the growth of microorganisms in the ground beef gel and extended its shelf-life without affecting the pH, cooking loss, color and texture characteristics of the product. In addition, the antibacterial effect of LCFS was the result of the interaction of organic acids and protein antibacterial substances in destroying cell structures (cell membrane, etc.) to achieve the purpose of bacteriostasis. This study provides a theoretical basis for the application of LCFS in meat products and a new clean-label strategy for the food industry.

Enhancing soil carbon and nitrogen through grassland conversion from degraded croplands in China: Assessing magnitudes and identifying key drivers of phosphorus reduction

Agricultural intensification has resulted in severe degradation of croplands in China. While the conversion of degraded croplands to grasslands has been proposed as a potential solution, the effects on soil organic carbon (SOC) and nutrients remain uncertain due to the complex interactions between plant growth and nutrient cycling. To identify key drivers and evaluate general patterns, a meta-analysis of 204 studies was conducted to evaluate changes in SOC, soil nitrogen (N), phosphorus (P), and soil enzyme activities in the 0–40 cm soil layer following grassland conversion across China. Our results showed that grassland conversion increased SOC (16 %), total N (TN, 12 %), available N (AN, 7 %), microbial biomass C (MBC, 48 %), microbial biomass N (MBN, 39 %), and the activities of urease (38 %) and phosphatase (64 %). However, the conversion decreased soil pH (−1 %) and available P content (AP, −26 %). The reduction in AP could be attributed to the restored grasslands absorbing more AP for growth. The duration of conversion was a major factor influencing changes in SOC, TN, and urease activities. Furthermore, mean annual precipitation had a significant impact on soil pH, TP, AP, MBC, and phosphatase activities. Our findings suggest that converting degraded croplands to grasslands can improve soil C and N content in China; however, these findings should be interpreted in the context of the entire recovery process. Further research and mitigation measures, such as the addition of soil P sources, may be required to address this decline in AP.

Targeting SIRT5 By Succinylating Hadha Synergizes with Venetoclax in Acute Myeloid Leukemia

Background: Acute myeloid leukemia (AML) is genetically diverse, driven by multiple oncogenic factors. Metabolic dysregulation is a critical vulnerability in AML cells for targeted therapy. BCL2 has been previously reported to maintain mitochondrial oxidative phosphorylation (OXPHOS) in AML cells. One of the mechanisms of action of the combination of venetoclax and azacitidine is to inhibit amino acid metabolism, leading to cell death. Although venetoclax-based treatment regimens have shown promising clinical outcomes in AML patients, treatment resistance has become a newly emerged challenge. Leukemia stem cells isolated from venetoclax-resistant patients exhibit distinctive metabolic features, including elevated nicotinamide metabolism. SIRT5 is an NAD+-dependent deacylase enzyme, and its knockout leads to elevated succinylation of various enzymes involved in OXPHOS. Hence, we hypothesize that inhibiting SIRT5 may augment venetoclax's cytotoxic effect by dampening metabolism. To further elucidate this mechanism, we employed proteome analyses of succinylation to identify specific target molecules regulated by SIRT5. Methods: To determine the effects of combination therapy on cell proliferation, apoptosis, and colony formation, AML cell lines were transduced with lentiviruses carrying either shNC or shSIRT5, or treated with NRD167 (a specific inhibitor of SIRT5), followed by venetoclax treatment. To evaluate the clearance of leukemia cells in vivo, NSG mice were transplanted with MOLM-13 cells (carrying shNC or shSIRT5) and then subjected to vehicle or venetoclax treatment (80 mg/kg/day) via oral gavage for 14 days. Weekly live animal in vivo imaging was conducted to monitor tumor engraftment and growth. The synergistic effect on the metabolic level of AML was assessed by measuring ATP, glutathione, mitochondrial superoxide, and oxygen consumption rates (OCR). Electron microscopy was employed to detect changes in cellular mitochondrial structure after SIRT5 knockdown and venetoclax utilization. Furthermore, lysine succinylation proteome analyses were performed to identify proteins and sites with altered succinylation on lysine before and after SIRT5 knockdown in MOLM-13 cells. Immunoprecipitation assays and Western blot were conducted to verify the interaction and altered succinylation levels of the relevant proteins. Results: The combination of SIRT5 inhibition with venetoclax treatment resulted in further suppression of cell proliferation, reduced colony formation, and increased apoptosis in AML cells compared to venetoclax treatment alone.The combination led to alterations in the metabolic profile of leukemia cells, increased the generation of mitochondrial reactive oxygen species (ROS), reduced intracellular glutathione levels, inhibited ATP production, and suppressed OCR. Notably, combination therapy resulted in a significant reduction in the engraftment levels of human leukemia cells in NSG mice. Mechanistically, electron microscopy revealed that combination treatment with SIRT5 knockdown increased mitochondrial damage. Lysine succinylation proteome analyses demonstrated upregulation of succinylation modification on mitochondrial trifunctional enzyme subunit α (HADHA), a protein involved in the fatty acid oxidation (FAO) pathway. Co-immunoprecipitation of SIRT5 confirmed its interaction with HADHA, and the relative succinylation level of precipitated HADHA was upregulated after SIRT5 knockdown. Moreover, the combination of trimetazidine (a specific inhibitor of HADHA) with venetoclax demonstrated a preferential reduction in cell proliferation and induction of apoptosis in AML cells. Conclusion: Targeting of SIRT5 in combination with venetoclax treatment results in synergistic cytotoxic effects in AML cells. We hypothesize that SIRT5 desuccinylase activity promotes FAO and reduces intracellular mitochondrial ROS levels by desuccinylating HADHA. Targeting SIRT5 inhibits fatty acid metabolism in AML cells, thereby enhancing the therapeutic effect of venetoclax. We will further investigate the site(s) of SIRT5 desuccinylation modification of HADHA through lysine site mutation and assess changes in HADHA enzyme activity to elucidate its regulatory mechanism.

Metabolism and Receptor Mechanisms of Niacin Action

The article discusses the metabolism of niacin, also known as vitamin B3 or PP, and the mechanisms of its receptor-induced functions in the human body. Niacin exists as a several molecular compounds that act as the nicotinamide coenzymes precursors. These coenzymes being electron donors or acceptors in redox reactions catalyzed by various enzymes play a crucial role in metabolism. Maintenance of the intracellular niacin pool is vital not only for redox metabolism, but also for the NAD-dependent pathways functioning. At the same time, pathophysiological situations and changes in enzyme activity can affect the necessity for various niacin forms. In addition to indirect effects via nicotinamide coenzymes, it also has a number of direct effects, including anti-lipolytic, vasodilatory, and neuroprotective functions, the exact mechanism of which has not been studied fully up to date. Overall, niacin plays a vital role in maintaining the efficient cell functioning, and further study of its influence on various physiological aspects, including the gut microbiome and epigenetic regulation, could lead to new discoveries and treatments for various diseases.

Microbial Community Shifts with Soil Properties and Enzyme Activities in Inter-/Mono-Cropping Systems in Response to Tillage

No-till and cereal–legume intercropping have been recognized as favorable cropping practices to increase crop yields while maintaining soil quality in arid and semiarid environments, but the biological mechanisms are poorly understood. The present study aimed to determine the response of yields, soil properties, enzyme activities, and microbial community diversity and composition in mono- and inter-cropping under conventional and no-tillage conditions. We initiated a field experiment in Wuwei, a typical arid area of China, in 2014. Soil was sampled in August 2022 and, yields, soil properties, enzyme activities, and the microbial community diversity and composition were determined in the maize and pea strips in inter- and mono-cropping systems. Results revealed that the maize and pea strips in the no-till intercropping significantly increased yields, total and organic carbon stocks, decreased NO3−-N, and obtained the highest total and organic P in the soil. No-tillage significantly enhanced the Shannon index and Pielou evenness of the bacterial community and total microbial community over conventional tillage, with the α-diversity of the bacterial community and total microbial community distinctly higher in the NTIM treatment than in the CTIM treatment. The α-diversity of the total microbial community was significantly related to yield, soil IC and OC, and the α-diversity of the archaea community was significantly related to soil TC, TC/TP, TN/TP, and BX. Meanwhile, the α-diversity of the eukaryote community was significantly related to soil yield, soil TC/TP. Both no-tillage and intercropped maize significantly increased the abundance of archaea phylum Thaumarchaeota and bacterial phylum Nitrospirae, and were significantly positively associated with soil OC and NH4+-N, benefiting nitrogen fixation of intercropped pea from the atmosphere under the no-tillage cereal/legume intercropping. No-till intercropping was conducive to the accumulation of organic carbon, while decreasing the abundance of Proteobacteria, Acidobacteria, and Verrucomicrobia. Limited soil enzyme activities (ACP, ALP, DP, NAG, BG, AG, CB) led to decreases in organic carbon turnover and utilization. Intercropping altered soil microbial community diversity and composition due to changes in soil properties and enzyme activities. These findings suggest that no-tilled cereal–legume intercropping is a sustainable cropping practice for improving soil properties and enhancing microbial (archaea, bacterial, eukaryota) diversity, but the persistence is not conducive to rapid turnover of soil nutrients due to limited enzyme activities.

The role of citrate synthase HbCS4 in latex regeneration of Hevea brasiliensis (para rubber tree)

The rubber tree, Hevea brasiliensis, is a tropical rubber-yielding crop with economic importance and provides over 90% of natural rubber (NR) for the global rubber industry. NR biosynthesis belongs to a typical secondary metabolism in laticifers. Definitely, NR production is a huge energy-consuming metabolism, especially during latex regeneration. The tricarboxylic acid (TCA) cycle, as the primary energy source, strongly supports NR synthesis in laticifers. In this study, a mitochondrial citrate synthase (CS) gene named as HbCS4 was isolated from rubber tree and related changes of expression patterns, enzyme activity and metabolites assays in response to tapping and ethephon (ethylene-releasing agent) treatments were compared. These molecular analyses were also conducted in two typical varieties Reyan 7–33–97 and PR107 as well as in HbCS4-overexpressing lines. As a result, the expression characteristics of HbCS4 or its enzyme activity tallied with the energy requirement of rubber synthesis in both treatments and the yielding traits of varieties Reyan 7–33–97 and PR107. In addition, in HbCS4-overexpressing lines, the activities of three key enzymes in TCA cycle, namely CS, NAD-IDH and α-KGDH, were all increased simultaneously. To conclude, these results suggested that HbCS4 is the initial enzyme required for energy supply in laticifers, which greatly contributes to rubber production through changing its expression or enzyme activity.

Synergistic Interactions in Enzyme Activities During Vermiremediation of Heavy Metals (HMs) Polluted Soil: Elucidating the Impact of Eudrilus eugeniae

ABSTRACT The vermiremediation potential of Eudrilus eugeniae was assessed in heavy metals (HMs) contaminated soil in a microcosm experiment. In detail, efficacy of E. eugeniae was investigated in terms of reduction in HMs content, enhancement in the soil enzyme activities, synergistic correlation between the HMs and enzyme activities and bioaccumulation of HMs in the earthworm’s biomass. Moreover, a seed germination assay was also conducted to assess the HMs toxicity in soil after remediation. The results showed that Eudrilus eugeniae was capable of decreasing the Cu, Zn, Mn, Cr, and Ni contents in the soil by 17.56–26.30% after 90 days of experimental trial. Introduction of E. eugeniae increased cellulase, amylase, polyphenol oxidase, peroxidase, urease, dehydrogenase, and catalase activities by 30.3–80.95% compared to controls. Earthworm addition led to a 12.98% increase in G-Mean and 2.89% increase in T-QSI values, indicating significant soil health improvement. PCA revealed the negative impact of HMs on synergistic enzyme activities during vermiremediation. HMs content in earthworm biomass increased 13-fold. Seed germination assay confirmed HMs reduction by the end of the experiment. Thus, this study demonstrated the interconnected changes in the soil enzyme activities during vermiremediation of HMs from crude oil polluted soil.

Vermiremediation of heavy metals (HMs)-contaminated agricultural land: synergistic changes in soil enzyme activities and earthworm's growth parameters.

In this investigation the heavy metals (Cu, Zn, Mn, Cr and Ni) remediation potential of Eisenia fetida was studied in the crude oil polluted soil. The potential of E. fetida was evaluated based on the decrease in concentrations of Cu, Zn, Mn, Cr and Ni, and improvement in the soil enzyme activities at the end of 90 days of experimental trials. Moreover, soil health quality, inter-relationship between the enzyme activities and the growth parameters of E. fetida and synergistic relation among the enzyme activities were also evaluated through G-Mean and T-QSI indices, chord plot analysis and principal component analysis (PCA) to confirm the performance of E. fetida during vermiremediation. The results revealed that the soil treated with E. fetida showed a reduction in the concentration of Cu, Zn, Mn, Cr and Ni by 17.4% 19.45%, 9.44%, 23.8% and 9.6% respectively by end of the experimental trials. The cellulase, amylase, polyphenol oxidase, peroxidase, urease, dehydrogenase and catalase activities in the E. fetida-treated soil were enhanced by 89.83%, 99.17%, 142%, 109.9%, 92.9%, 694.3% and 274.5% respectively. The results of SEM-EDS revealed enhancement in the O, K, Na, Mg and P content by 62.36%, 96.2%, 97.9%, 93.7% and 98.2% respectively by the end of the experimental trial. The G-Mean and T-QSI indices also confirmed the improvement in soil enzyme activities thereby indicating the positive influence of E. fetida on soil decontamination process. The chord plot indicated the interrelationship between the earthworm's growth parameters and enzyme activities of the soil as indicated by the high linkage between the nodes. Finally, the PCA confirmed the negative effect of the heavy metals on the soil enzyme activities and synergistic interrelationship between the enzyme activities during the vermiremediation process. Thus, this study demonstrated the changes in the soil enzyme activities and their interconnected influences during vermiremediation of crude oil sourced heavy metals from polluted soil.

Changes in Nutrient Components and Digestive Enzymatic Inhibition Activities in Soy Leaves by Ethephon Treatment.

In this study, the high isoflavone-enriched soy leaves (IESLs) were manufactured by treating with the chemical inducer ethephon, a plant growth regulator, to confirm changes in the properties of soy leaves (SLs), which are underutilized. Ethephon treatment concentrations consisted of 0 (SL1), 150 (SL2), and 300 (SL3) μg/mL. The composition analysis and physiological activity were conducted according to the ethephon treatment concentration of SLs. There was no significant difference in the proximate composition and fatty acids, except for an increase with increasing ethephon treatment concentrations. Depending on the ethephon treatment concentration, free amino acids increased to 1413.0, 1569.8, and 2100.4 mg/100 g, and water-soluble vitamins increased to 246.7, 244.7, and 501.6 mg/100 g. In particular, the functional substance isoflavone increased significantly to 1430.11, 7806.42, and 14,968.00 μg/g. Through this study, it was confirmed that the nutritional components and isoflavones of SLs increased according to the ethephon treatment concentration, a chemical inducer treatment agent. This can be used as a high-value-added biosubstance for raw materials for functional foods, cosmetics, and for natural drugs.

The deleterious effects of cadmium on oxidative stress markers, drug-metabolizing, and antioxidant enzyme activities: Role of Silymarin and Garlic as Antioxidants.

Exposure to cadmium has been related to liver and kidney diseases such as polycystic and nephrotic syndrome. It is still unclear how cadmium contributes to these diseases. It is believed that the induction of oxidative stress resulting from the inhibition of antioxidant enzyme activities and changes in drug-metabolizing enzymes in the liver could explain the role of cadmium in the development of different diseases in the kidney and probably other organs. Changes in oxidative stress markers, antioxidant enzymes, and drug-metabolizing enzyme activities were assessed in the liver of male rats exposed to cadmium chloride. Additionally, the protective effects of silymarin and garlic extract against cadmium toxicosis were evaluated. Rats were randomly divided into eight groups as follows, groups 1, 2, 3, 4, and 5, received orally saline, CdCl2 (1mg/kg), garlic extract [800mg/kg], silymarin (25mg/kg) and silymarin plus garlic extract respectively for 28 consecutive days. Rats in groups 6, 7, and 8 were pretreated with the same doses of garlic, silymarin, and garlic plus silymarin, respectively for two hours before cadmium administration. The Western immunoblotting technique was used to investigate the protein expression of cytochrome P450 isozymes. Spectrophotometric methods were used to assess the activity of both antioxidant- and drug-metabolizing enzymes. Free radical levels [measured as thiobarbituric acid reactive substances (TBARS)], catalase, superoxide dismutase, and glutathione peroxidase activities increased whereas the levels of glutathione and the activities of glutathione S-transferase, glutathione reductase, and glutamyl transferase, cytochrome P450, aryl hydrocarbon dehydrogenase (AHH), dimethylnitrosamine-N-demethylase I (DMN-dI), 7-ethoxycoumarine-O-deethylase (ECOD), cytochrome b5 and NADPH-Cytochrome-c-reductase enzyme activities decreased after cadmium treatment. Furthermore, Western immunoblotting data revealed that glutathione peroxidase protein expression increased following cadmium exposure, but cytochrome P450 2E1 and 3A4 expressions were downregulated. However, pretreatment of rats with silymarin or garlic extract or both before cadmium administration was found to restore the protein expression of cytochrome P450 2E1 and 3A4, the level of free radicals, antioxidant enzymes, drug-metabolizing enzyme activities to their normal levels. Similarly, histological studies revealed that silymarin and/or garlic extract reduced the liver damage caused by cadmium. Silymarin and/or garlic extract reduced the adverse effects of cadmium on the activity of both drug-metabolizing and antioxidant enzymes activity. These antioxidants could be provided to those who work in cadmium-based sectors to help them cope with the adverse effects of cadmium on their kidneys. In addition, Inhibiting drug-metabolizing enzyme activity should be considered when administering therapeutic medications to persons exposed to cadmium because most therapeutic drugs and many endogenous substances are largely metabolized by these enzymes.

Long-Term Nitrogen Addition Could Modify Degradation of Soil Organic Matter through Changes in Soil Enzymatic Activity in a Natural Secondary Forest

Soil extracellular enzymes play a key role in mediating the degradation of soil organic matter, but little is understood as to how the pattern of soil extracellular enzymes could be altered by nitrogen (N) addition. In this study, the effects of N addition (started from 2006, four treatments: control, 0 g N·m–2·yr–1; low N addition, 2.5 g N·m–2·yr–1; high N addition, 5.0 g N·m–2·yr–1) on soil extracellular enzymes and microbial biomass in a natural secondary forest of Northeastern China. The results showed that the activity of urease, sucrase, peroxidase and N-acetyl-β-D-glucosidase decreased with N addition, and the activity of acid phosphatase and leucine aminopeptidase increased significantly with N addition. Soil total N, temperature at 5 cm soil depth, pH value, microbial biomass carbon and microbial biomass N were the key factors affecting soil enzyme activity. In summary, the enzyme activity related to soil organic matter degradation shows a decreasing trend under N addition. The results suggest that the increase in N deposition will slow down the degradation of soil organic matter in natural secondary forest, which is more conducive to the accumulation of organic matter in the soil.

Response of maize stalk to plant density on cellulose accumulation by modulating enzymes activities

Hybrids and plant density are the main factors affecting maize lodging resistance, and cellulose plays an important role in lodging resistance. High plant density often decreases the stalk cellulose, but the cause is unclear. Therefore, we hypothesized that enzymatic activity might be the main factor affecting maize cellulose accumulation. For testing this hypothesis, three plant densities （4.5, 9.0 and 13.5 plants m−2） were set up and three maize hybrids （high lodging resistant cultivars: Xianyu 335, Zhengdan 958; low lodging resistant cultivar: Xinyu 41）with different lodging resistance were selected as materials. The results showed that high plant densities reduced the stalk cellulose contents and strength, and hence lodging risk increased. Through logistic curve fitting, we found that the period of rapid accumulation of cellulose in stalks was 10–26 DPV6 (days post 6-leaf stage). Analyse changes of stalk sucrose and related enzyme activity during 10–26 DPV6 showed that with the increase of plant density, the sucrose content of the stalks significantly decreased (P＜0.05), sucrose content began to differ at 16-DPV6. Both sucrose phosphate synthase(SPS) and sucrose synthase(SS) activity of stalks in this stage decreased with the increase of plant densities. At 20 DPV6, there has a great difference in SPS among different plant densities during this period. Compared with 4.5 plants m−2, the decreases for SPS in Xianyu 335 and Zhengdan 958 was lower than Xinyu 41 at 9.0 plants m−2 and 13.5 plants m−2. The SPS for Xianyu 335 decreased by 10.0%− 16.0% and 28.7%− 37.7% at 9.0 plants m−2 and 13.5 plants m−2, respectively. However, the SPS for Xinyu 41 decreased by 24.4%− 27.3% and 40.2%− 50.2%, respectively. At 16 DPV6, there was a large difference in SS among different plant densities, the decreases for SS in Xianyu 335 and Zhengdan 958 was lower than Xinyu 41. Xianyu 335 decreased by 3.6%− 15.9%, 18.8%− 32.7% at at 9.0 plants m−2 and 13.5 plants m−2, respectively. Similarly Xinyu 41 decreased by 18.2%− 26.4% and 31.8%− 50.6%, respectively. Thus, under high plant density, the activities of SS and SPS in maize stalks were decreased, the accumulation and conversion of sucrose into cellulose were inhibited, and the activity of cellulose synthesis-related enzymes in varieties with poor lodging resistance were more sensitive to the increase of density. Therefore, we concluded that higher activity of SS and SPS increased cellulose accumulation and stalk strength in maize varieties with better lodging resistance reduced the lodging percentage under close density conditions.

Acute thermal stress increased enzyme activity and muscle energy distribution of yellowfin tuna.

Heat is a powerful stressor for fish living in natural and artificial environments. Understanding the effects of heat stress on the physiological processes of fish is essential for better aquaculture and fisheries management. In this experiment, a heating rod was used to increase the temperature at 2°C/h to study the changes of energy allocation (CEA) and energy metabolity-related enzyme activities, including pepsin, trypsin, amylase, lipase, acid phosphatase, lactate dehydrogenase, alanine aminotransferase, glutamic oxalic aminotransferase and energy reserve (Ea), energy expenditure (ETS), in juvenile yellowfin tuna cells under acute temperature stress. The results showed that the Ea of juvenile yellowfin tuna muscles in response to high temperature (34°C) was significantly lower than that of the control (28°C), and it also increased ETS. At 6 h, CEA decreased slightly in the high-temperature group, but, the difference in CEA between 24 h and 0 h decreased. After heat stress for 6 h, the activities of acid phosphatase (ACP), lactate dehydrogenase (LDH), alanine aminotransferase (ALT) and glutamic oxalacetic transaminase (AST) increased, indicating that the metabolic rate was accelerated. After heat stress for 24 h, the activity of ALT decreased, indicating that with time elapsed, the activities of some protein metabolizing enzymes increased, and some decreased. In this study, digestive enzymes, trypsin and lipase increased gradually. After heat stress, Ea and Ec change significantly. Yellowfin tuna muscles use lipids in response to sharp temperature increases at high temperatures, red muscles respond to temperature changes by increasing energy in the early stages, but not nearly as much, and white muscles reduce lipids.

Butyl benzyl phthalate exposure impact on the gut health of Metaphire guillelmi

Agricultural films are extensively utilized in high-intensity agriculture, with China's annual usage reaching 1.5 million tons. Unfortunately, the recovery rate is less than 60%, leading to an inevitable accumulation of plastic mulch in agricultural soils. This accumulation primarily introduces butyl benzyl phthalate (BBP) into soil ecosystems, whose specific effects remain largely unclear, thereby posing potential risks. The present study focuses on the exposure impact of BBP on earthworms, Metaphire guillelmi, a commonly found endogenic earthworm within real farmland, as it provides insight into the direct interaction between biota gut health and contaminants. Specifically, we studied the biomarkers related to oxidative stress, the digestive system, and neurotoxicity within the gut of Metaphire guillelmi, and the integrated biological response (IBR) index was utilized to track these markers at different timeframes after BBP exposures. Our findings indicate that BBP exposures lead to oxidative damage, digestive system inhibition, and neurotoxicity, with IBR indexes of 14.6 and 17.3 on the 14th and 28th days, respectively. Further, the underlying mechanisms at a molecular level through molecular docking were investigated. The results showed that the most unstable interaction was with the Na+K+-ATPase (binding energy: −2.25 kcal−1), while BBP displayed stable bonds with superoxide dismutase and 8-hydroxydeoxyguanosine via hydrogen bonds and hydrophobic interaction. These interactions resulted in changes in protein conformation and their normal physiological functions, offering new insights into the molecular mechanism underlying enzymatic activity changes. This study has significant implications for the prediction of toxicity, environmental risk assessment, and the establishment of regulations related to BBP.

Intrinsic structural dynamics dictate enzymatic activity and inhibition

Enzymes are known to sample various conformations, many of which are critical for their biological function. However, structural characterizations of enzymes predominantly focus on the most populated conformation. As a result, single-point mutations often produce structures that are similar or essentially identical to those of the wild-type enzyme despite large changes in enzymatic activity. Here, we show for mutants of a histone deacetylase enzyme (HDAC8) that reduced enzymatic activities, reduced inhibitor affinities, and reduced residence times are all captured by the rate constants between intrinsically sampled conformations that, in turn, can be obtained independently by solution NMR spectroscopy. Thus, for the HDAC8 enzyme, the dynamic sampling of conformations dictates both enzymatic activity and inhibitor potency. Our analysis also dissects the functional role of the conformations sampled, where specific conformations distinct from those in available structures are responsible for substrate and inhibitor binding, catalysis, and product dissociation. Precise structures alone often do not adequately explain the effect of missense mutations on enzymatic activity and drug potency. Our findings not only assign functional roles to several conformational states of HDAC8 but they also underscore the paramount role of dynamics, which will have general implications for characterizing missense mutations and designing inhibitors.