Screening for metabolic dysfunction-associated fatty liver disease:Time to discard the emperor’s clothes of normal liver enzymes?

Metabolic dysfunction-associated fatty liver disease(MAFLD)is the most prevalent chronic liver condition worldwide.Current liver enzyme-based screening methods have limitations that may missed diagnoses and treatment delays.Regarding Chen et al,the risk of developing MAFLD remains elevated even when alanine aminotransferase levels fall within the normal range.Therefore,there is an urgent need for advanced diagnostic techniques and updated algorithms to enhance the accuracy of MAFLD diagnosis and enable early intervention.This paper proposes two potential screening methods for identifying individuals who may be at risk of developing MAFLD:Lowering these thresholds and promoting the use of noninvasive liver fibrosis scores.

The Effect of Soil Enzymes and Polysaccharides Secreted by the Roots of Salvia miltiorrhiza Bunge under Drought,High Temperature,and Nitrogen and Phosphorus Deficits

Root exudates serve as crucial mediators for information exchange between plants and soil,and are an important evolutionary mechanism for plants’adaptation to environmental changes.In this study,15 different abiotic stress models were established using various stress factors,including drought(D),high temperature(T),nitrogen deficiency(N),phosphorus deficiency(P),and their combinations.We investigated their effects on the seedling growth of Salvia miltiorrhiza Bunge and the activities of Solid-Urease(S-UE),Solid-Nitrite Reductase(S-NiR),Solid-Nitrate Reductase(S-NR),Solid-Phosphotransferase(S-PT),and Solid-Catalase(S-CAT),as well as the contents of polysaccharides in the culture medium.The results showed that the growth of S.miltiorrhiza was inhibited under 15 stress conditions.Among them,13 stress conditions increased the root-shoot ratio.These 15 stress conditions significantly reduced the activity of S-NR,two combinations significantly improved the activity of S-NIR,they were synergistic stresses of high temperature and nitrogen deficiency(TN),and synergistic stresses of drought and nitrogen deficiency(DN)(p<0.05).The activity of S-UE was significantly improved under N,D,T,synergistic stresses of drought and high temperature(DT),DN,synergistic stresses of drought and phosphorus deficiency(DP),and synergistic stresses of high temperature,nitrogen,and phosphorus deficiency(TNP)stress conditions(p<0.05).Most stress combinations reduced the activity of S-PT,but D and T significantly improved it.(p<0.05).The N,DN,and TN stress conditions significantly reduced S-CAT activity.The P,DT,and synergistic stresses of drought,high temperature,and phosphorus deficiency(DTP)significantly decreased the total polysaccharide content of the soil(p<0.05).The research suggested that abiotic stress hindered the growth of S.miltiorrhiza and altered the behavior of root secretion.Roots regulated the secretion of several substances in response to various abiotic stresses,including soil nitrogen cycle enzymes,phosphorus transport-related enzymes,and antioxidant enzymes.In conclusion,plants regulate the utilization of rhizosphere substances in response to abiotic stresses by modulating the exudation of soil enzymes and polysaccharides by the root system.At the same time,soil carbon sequestration was affected by the adverse environment,which restricted the input of organic matter into the soil.

Fat,oil,and grease as new feedstock towards bioelectrogenesis in microbial fuel cells:Microbial diversity,metabolic pathways,and key enzymes

Microbial fuel cells(MFCs)are a well-known technology used for bioelectricity production from the decomposition of organic waste via electroactive microbes.Fat,oil,and grease(FOG)as a new substrate in the anode and microalgae in the cathode were added to accelerate the electrogenesis.The effect of FOG concentrations(0.1%,0.5%,1%,and 1.5%)on the anode chamber was investigated.The FOG degradation,volatile fatty acid(VFAs)production,and soluble chemical oxygen demand along with voltage output kinetics were analyzed.Moreover,the microbial community analysis and active functional enzymes were also evaluated.The maximum power and current density were observed at 0.5%FOG which accounts for 96 mW m^(-2)(8-folds enhancement)and 560 mA m^(-2)(3.7-folds enhancement),respectively.The daily voltage output enhanced upto 2.3-folds with 77.08%coulombic efficiency under 0.5%FOG,which was the highest among all the reactors.The 0.5%FOG was degraded>85%,followed by a 1%FOG-loaded reactor.The chief enzymes inβ-oxidation and electrogenesis were acetyl-CoA C-acetyltransferase,riboflavin synthase,and riboflavin kinase.The identified enzymes symbolize the presence of Clostridium sp.(>15%)and Pseudomonas(>10%)which served as electrochemical active bacteria(EAB).The major metabolic pathways involved in electrogenesis and FOG degradation were fatty acid biosynthesis and glycerophospholipid metabolism.Utilization of lipidic-waste(such as FOG)in MFCs could be a potential approach for simultaneous biowaste utilization and bioenergy generation.

Peptide self‐assembly as a strategy for facile immobilization of redox enzymes on carbon electrodes

Redox-enzyme‐mediated electrochemical processes such as hydrogen production,nitrogen fixation,and CO_(2) reduction are at the forefront of the green chemistry revolution.To scale up,the inefficient two‐dimensional(2D)immobilization of redox enzymes on working electrodes must be replaced by an efficient dense 3D system.Fabrication of 3D electrodes was demonstrated by embedding enzymes in polymer matrices.However,several requirements,such as simple immobilization,prolonged stability,and resistance to enzyme leakage,still need to be addressed.The study presented here aims to overcome these gaps by immobilizing enzymes in a supramolecular hydrogel formed by the self‐assembly of the peptide hydrogelator fluorenylmethyloxycarbonyldiphenylalanine.Harnessing the self‐assembly process avoids the need for tedious and potentially harmful chemistry,allowing the rapid loading of enzymes on a 3D electrode under mild conditions.Using the[FeFe]hydrogenase enzyme,high enzyme loads,prolonged resistance against electrophoresis,and highly efficient hydrogen production are demonstrated.Further,this enzyme retention is shown to arise from its interaction with the peptide nanofibrils.Finally,this method is successfully used to retain other redox enzymes,paving the way for a variety of enzyme‐mediated electrochemical applications.

Toxic Effects of Citrus maxima Based Combinatorial Formulations on Important Metabolic Enzymes in Indian White Termite Odontotermes obesus

In the present study, Citrus maxima essential crude oil extract was used to prepare combinatorial formulations and workers of Indian white termite Odontotermes obesus were treated topically with 40% and 80% of 24 hr LD50 values of these formulations. In subsequent bioassays levels of various enzymes i.e. alkaline phosphatase, acid phosphatase, glutamate oxaloacetate transaminase, glutamate pyruvate transaminase and acetylcholinesterase were evaluated to determine the anti-termite efficacy of Citrus maxima essential oil based combinatorial formulations. S-RET-A, S-RET-B and S-RET-C caused significant (p > 0.05) decrease in glutamate oxaloacetate transaminase i.e. 87.47%, 86.81% and 81.77% & 82.04%, 79.39% and 74.75% respectively at 16 h treatment. In vivo exposure of 40% and 80% of LD50 of combinatorial formulations caused very significant (p > 0.05) reduction in all the test enzymes i.e. alkaline phosphatase, acid phosphatase, glutamate oxaloacetate transaminase and glutamate pyruvate transaminase expect acetylcholinesterase levels after 16 h all tested treatments in comparison to control. Both dose-response and time period were found important in physiological alteration in levels of various enzymes. Combinatorial mixtures of Citrus essential oils have shown synergistic activity against termites. The research findings of the present study would help termite control in crop fields, gardens and houses in a sustainable way.

Toxicity of Tagetes erecta Essential Oil Based Combinatorial Formulations on Various Metabolic Enzymes in Indian White Termite Odontotermes obesus

Plant essential oils and their constituents have proven to be very effective against insects, especially termites. They are the best alternative to synthetic pesticides that are harmless to the environment and human health. In the present study, different enzymes, namely alkaline phosphatase, acid phosphatase, glutamate-oxaloacetate transaminase, glutamate-pyruvate transaminase and acetylcholinesterase, were evaluated in Indian termite Odontotermes obesus in a combination preparation based on Tagetes erecta essential oil. For the study of anti-termite effects against worker termites were treated topically with 40% and 80% of the 24-hour LD50 values of various combination formulations. Subsequent bioassays at 40% and 80% of LD50 of combinatorial preparations S-AST-A, S-AST-B, and S-AST-C resulted in a significant (p > 0.05) decrease in glutamate-oxaloacetate transaminase. After 16 hours of treatment, they were 87.92%, 80.17%, 89.30%, 79.17%, 81.92% and 73.17% respectively. In vivo exposures of 40% and 80% of the LD50 of the combination formulation caused a highly significant (p > 0.05) reduction (p > 0.05) of all test enzymes tested compared to controls. The effects of different oil formulations exhibited time- and dose-dependent responses, resulting in physiological changes in the concentrations of various enzymes. The combined mixture of Tagetes erecta essential oils has significantly better anti-termite ability compared to inorganic insecticides. Findings from this study will help support termite control in fields, gardens and homes in a sustainable way, without the downsides of insecticide resistance and pollution. These could potentially be used to produce commercial formulations for use against pests.

Investigation and Benefits of the Use of Enzymes for Unbleached Kraft Recycling Wet Strength Paper at Lab and Industrial Scale

The deterioration of recycled fibers especially unbleached kraft with high wet strength resin content due to the irreversible structural changes caused by drying and added chemicals makes the raw material difficult to repulp.The mechanical effect in the pulper over time with chemical treatment has a negative impact on the recycled fibers.At lab scale,different compositions of enzymatic treatment C022L were under investigation to select the most efficient laccase Lacc1,Lacc2 or Lacc3 and to observe the impact of lipases during repulping at low and high consistencies.Pulp disintegration at different times was evaluated to define the level of rejects and to analyze the morphology of fibers after treatments.These results were more significant for Lacc2,by increasing the pulp consistency to 15%in the pulper.Combining lipases with CELODASE 022L appeared to decrease the efficiency of enzymes.The results showed a high reduction of energy power with the enzymatic treatment and a significant reduction of fines level in fibers’suspension.The most efficient version of C022L was used at industrial scale to compare directly with the standard conditions used in a paper mill.

Plant-Derived Enzymes Producing Chiral Aroma Compounds and Potential Application

Aroma(volatile)compounds play important ecological functions in plants,and also contribute to the quality of plant-derived foods.Moreover,chiral aroma compounds affect their functions in plants and lead to different flavor quality properties.Formations of chiral aroma compounds are due to the presence of enzymes producing these compounds in plants,which are generally involved in the final biosynthetic step of the aroma compounds.Here,we review recent progress in research on the plant-derived enzymes producing chiral aroma compounds,and their changes in response to environmental factors.The chiral aroma enzymes that have been reported produce(R)-linalool,(S)-linalool,(R)-limonene,and(S)-limonene,etc.,and these enzymes are found in various plant species.We also discuss the origins of enantioselectivity in the plant-derived enzymes producing chiral aroma compounds and summarize the potential use of plants containing enzymes producing chiral aroma compounds for producing chiral flavors/fragrances.

Changes in the activities of key enzymes and the abundance of functional genes involved in nitrogen transformation in rice rhizosphere soil under different aerated conditions

Soil microorganisms play important roles in nitrogen transformation. The aim of this study was to characterize changes in the activity of nitrogen transformation enzymes and the abundance of nitrogen function genes in rhizosphere soil aerated using three different methods(continuous flooding(CF), continuous flooding and aeration(CFA), and alternate wetting and drying(AWD)). The abundances of amoA ammonia-oxidizing archaea(AOA) and ammonia-oxidizing bacteria(AOB), nirS, nirK, and nifH genes, and the activities of urease, protease, ammonia oxidase, nitrate reductase, and nitrite reductase were measured at the tillering(S1), heading(S2), and ripening(S3) stages. We analyzed the relationships of the aforementioned microbial activity indices, in addition to soil microbial biomass carbon(MBC) and soil microbial biomass nitrogen(MBN), with the concentration of soil nitrate and ammonium nitrogen. The abundance of nitrogen function genes and the activities of nitrogen invertase in rice rhizosphere soil were higher at S2 compared with S1 and S3 in all treatments. AWD and CFA increased the abundance of amoA and nifH genes, and the activities of urease, protease, and ammonia oxidase, and decreased the abundance of nirS and nirK genes and the activities of nitrate reductase and nitrite reductase, with the effect of AWD being particularly strong. During the entire growth period, the mean abundances of the AOA amoA, AOB amoA, and nifH genes were 2.9, 5.8, and 3.0 higher in the AWD treatment than in the CF treatment, respectively, and the activities of urease, protease, and ammonia oxidase were 1.1, 0.5, and 0.7 higher in the AWD treatment than in the CF treatment, respectively. The abundances of the nirS and nirK genes, and the activities of nitrate reductase and nitrite reductase were 73.6, 84.8, 10.3 and 36.5% lower in the AWD treatment than in the CF treatment, respectively. The abundances of the AOA amoA, AOB amoA, and nifH genes were significantly and positively correlated with the activities of urease, protease, and ammonia oxidase, and the abundances of the nirS and nirK genes were significantly positively correlated with the activities of nitrate reductase. All the above indicators were positively correlated with soil MBC and MBN. In sum, microbial activity related to nitrogen transformation in rice rhizosphere soil was highest at S2. Aeration can effectively increase the activity of most nitrogen-converting microorganisms and MBN, and thus promote soil nitrogen transformation.

Enhanced production of cytidine 5'-monophosphate using biocatalysis of di-enzymes immobilized on amino-functionalized sepharose

Cytidine 5'-monophosphate(5'-CMP)is an essential nucleotide for additives.In this study,enhanced production of 5'-CMP was realized by the transformation of cytidine using co-immobilized di-enzymes,uridine-cytidine kinase(UCK)and acetate kinase(AcK).The immobilization yield of the enzyme had a clear correlation with the surface charges as zeta potential(ξ).Among them,ε-polylysinefunctionalized sepharose(SA-EPL,ξ=9.31 m V)showed high immobilization yield(78.8%),which was4.9-fold than that of nitrilotriacetic acid functionalized sepharose(SA-NTA,ξ=-12.6 m V).The residual activity of affinity co-immobilized enzyme(EPL-Ni/EPL@Ac K-UCK)was higher than 70.6%after recycled 10 times.Thus,this study provides an effective approach for the production of 5'-CMP with the advantages of low adenosine 5'-triphosphate(ATP)consumption,reduced side reactions,and improved reusability by co-immobilized UCK and Ac K on the functionalized Sepharose.

Effects of Different Nitrogen and Phosphorus Synergistic Fertilizer on Enzymes and Genes Related to Nitrogen Metabolism in Wheat

In recent years,in order to improve nutrient use efficiency,especially nitrogen use efficiency,fertilizer valueadded technology has been developed rapidly.However,the mechanism of the effect of synergistic fertilizer on plant nitrogen utilization is not clear.A study was,therefore,conducted to explore the activities and gene expression of key enzymes for nitrogen assimilation and the gene expression of nitrogen transporters in wheat after the application of synergistic fertilizer.Soil column experiment was set up in Qingdao Agricultural University experimental base from October 2018 to June 2019.Maleic acid and itaconic acid were copolymerized with acrylic acid as cross-linking monomer to make a fluid gel,which was sprayed on the fertilizer surface to make nitrogen and phosphorus synergistic fertilizer.A total of 6 treatments was set according to different nitrogen and phosphorus fertilizer ratios:(1)100%common nitrogen fertilizer+100%common phosphate fertilizer(2)70%nitrogen synergistic fertilizer+100%phosphorus synergistic fertilizer;(3)100%nitrogen synergistic fertilizer+70%phosphorus synergistic fertilizer;(4)100%nitrogen synergistic fertilizer+100%phosphorus synergistic fertilizer;(5)70%nitrogen synergistic fertilizer+70%phosphorus synergistic fertilizer;(6)100%commercial nitrogen synergistic fertilizer+100%commercial phosphorus synergistic fertilizer.The results are as follows:(1)the enzyme activities of wheat plants under synergistic fertilizer condition were higher than those under ordinary fertilizer,except under the treatment that nitrogen and phosphorus synergistic fertilizer were both reduced;(2)the expression level of the genes under the treatment“100%nitrogen synergistic fertilizer+100%phosphorus synergistic fertilizer”was significantly higher than those in other treatments.Combined with the higher performance of nitrogen concentration in various parts of the plant under the condition of applying synergistic fertilizer,this study indicated that the application of synergistic fertilizer can improve the nitrogen metabolism of the plant by increasing the nitrogen level in the rhizosphere soil,inducing the expression of nitrogen transporter genes and key assimilation enzymes genes.

Effect of Cellulose Enzymes on Some Properties of Fibre Clothes

This study examined the effects of the enzyme cellulose on a few single jersey and interlock 100% flax t-shirts, two different styles of knit children’s clothes that differ in both their chemical and mechanical characteristics. Clothing samples were dyed using reactive colours and then subjected to normal acid cellulose enzyme treatment procedures. We evaluated some physical and mechanical features before and after cellulose treatments, then compared these characteristics. Its cellulose enzymatic processing enhances some mechanical properties of fibre knitwear, such as pilling resistance and retention of water. Some characteristics of fibre-knitted clothing, such as fabric weight, fabric thickness, fabric burst resistance, and seam tensile strength of T-shirt side seams, are reduced by cellulose enzyme treatment at a manageable rate. Compared to single-jersey all-fiber children’s T-shirts, interlocking 100% flax children’s T-shirts have a better effect of the enzyme cellulose treatment on the majority of physical and mechanical attributes.

Changes of Proline Content,Activity,and Active Isoforms of Antioxidative Enzymes in Two Alfalfa Cultivars Under Salt Stress

The plants of two elfalfa （Medicago sativa L.） cultivars differing in salt tolerance were subjected to three salt treatments, 70, 140, and 210 mM NaCl for 7 days. Root, shoot, and leaf growths were inhibited by increased salt treatments in both cultivars, and at 140 and 210 mM salt treatments, Zhongmu 1 had significantly higher root, shoot, and leaf dry weights per plant than Deft. The malondialdehyde （MDA） accumulation in Deft was considerably greater than in Zhongmu 1, indicating a higher degree of lipid peroxidation at 140 and 210 mM salt treatments. The changes in the activity and active isoforms of antioxidant enzymes such as superoxide dismutase （SOD, EC 1.15.1.1）, catalase （CAT, EC 1.11.1.6）, peroxidase （POD, EC 1.11.1.7）, and ascorbate peroxidase （APOX, EC 1.11,1.11）, accumulation of free proline, and rate of lipid peroxidation in leaves of two alfalfa cultivars were also investigated. After stress, the activity and active isoforms of antioxidative enzymes were altered and the extent of alteration varied between the cultivar Deft and Zhongmu 1. The proline accumulation in Deft was considerably greater than in Zhongmu 1 at 210 mM salt treatment. This indicated that proline accumulation may be the result, instead of the cause, of salt tolerance.

Effects of melatonin on lipid peroxidation and antioxidant enzymes in streptozotocin-induced diabetic rat testis

Aim： To examine the effects of melatonin treatment on lipid peroxidation （LPO） and the activities of antioxidant enzymes in the testicular tissue of streptozotocin （STZ）-induced diabetic rats. Methods： Twenty-six male rats were randomly divided into three groups as follows： group Ⅰ, control, non-diabetic rats （n = 9）; group Ⅱ, STZ-induced, untreated diabetic rats （n = 8）; group Ⅲ, STZ-induced, melatonin-treated （dose of 10 mg/kg·day） diabetic rats （n = 9）. Following 8-week melatonin treatment, all rats were anaesthetized and then were killed to remove testes from the scrotum. Results： As compared to group Ⅰ, in rat testicular tissues of grouap Ⅱ, increased levels of malondialdehyde （MDA） （P 〈 0.01） and superoxide dismutase （SOD） （P 〈 0.01） as well as, decreased levels of catalase （CAT） （P 〈 0.01） and glutathione peroxidase （GSH-Px） （P 〉 0.05） were found. In contrast, as compared to group Ⅱ, in rat testicular tissues of group Ⅲ, levels of MDA decreased （but this decrease was not significant, P 〉 0.05） and SOD （P 〈 0.01） as well as CAT （P 〈 0.05） increased. GSH-Px was not influenced by any of the treatment. Melatonin did not significantly affect the elevated glucose concentration of diabetic group. At the end of the study, there was no significant difference between the melatonin-treated group and the untreated group by means of body and testicular weight. Conclusion： Diabetes mellitus increases oxidative stress and melatonin inhibits lipid peroxidation and might regulate the activities of antioxidant enzymes of diabetic rat testes.

Effects of insoluble Zn,Cd,and EDTA on the growth,activities of antioxidant enzymes and uptake of Zn and Cd in Vetiveria zizanioides

A root-bag experiment was conducted to study the effects of insoluble Zn, Cd, and ethylenediaminetetraacetic acid （EDTA） on the plant growth, activities of antioxidant enzymes, proline, glutathione （GSH）, water-soluble proteins and malondialdehyde （MDA） of Vetiveria zizanioides. The V. zizanioides uptake capacity of Zn and Cd also determined. The results showed that plant growth of V. zizanioides was inhibited by Zn and Cd. The shoot dry weight （SDW） and root dry weight （RDW） decrease by 14.2%, 14.1%, 17.0% and 17.3%, 32.5%, 35.7%, respectively, compared to the control without EDTA addition. After adding EDTA, shoot and root dry weight decreased over 10% and 15%, respectively. The toxicity from insoluble Zn and Cd in soil on SDW and RDW of V. zizanioides was in order： Zn＋Cd 〉 Cd 〉 Zn. The activities of superoxide dismutase （SOD）, peroxidase （POD） and catalase （CAT）, and contents of MDA and proline increased significantly, while the contents of GSH and water-soluble proteins decreased markedly with increasing Zn and Cd toxicity. With EDTA, shoot and root Zn concentrations increased in the Zn treatment by 7.3% and 37.4%, and Cd concentrations in the combined Zn and Cd treatment increased by 18.6% and 391.9% compared to the treatment without EDTA. However, Zn and Cd concentrations in shoot and roots decreased in the Cd treatment compared to the plants grown in absence of EDTA, with exception of root Cd concentration in the presence of EDTA.

Changes of Antioxidative Enzymes and Lipid Peroxidation in Leaves and Roots of Waterlogging-Tolerant and Waterlogging-Sensitive Maize Genotypes at Seedling Stage

To better understand the physiological and biochemical mechanisms of waterlogging tolerance, waterlogging effects on lipid peroxidation and the activity of antioxidative enzymes were investigated in leaves and roots of two maize genotypes, HZ32 （waterlogging-tolerant） and K12 （waterlogging-sensitive）. Potted maize plants were waterlogged at the second leaf stage under glasshouse conditions. Leaves and roots were harvested 1 d before and 2, 4, 6, 8 and 10 d after the start of waterlogging treatment. Through comparing the activities of superoxide dismutase （SOD）, ascorbate peroxidase （APX）, glutathione reductase （GR）, catalase （CAT） and guaiacol peroxidase （POD） between waterlogging-tolerant and waterloggingsensitive genotype, we deduced that CAT was the most important H2O2 scavenging enzyme in leaves, while APX seemed to play a key role in roots. POD, APX, GR and CAT activities in conjunction with SOD seem to play an essential protective role in the O2^- and H2O2 scavenging process. Lipid peroxidation was enhanced significantly only in K12 （P 〈 0.001） and there was no difference （P 〉 0.05） in HZ32 up to 6 d after waterlogging stress. These results indicated that oxidative stress may play an important role in waterlogging-stressed maize plants and that the greater protection of HZ32 leaves and roots from waterlogging-induced oxidative damage results, at least in part, through the maintenance of increased antioxidant enzyme activity.

Changes of Reactive Oxygen Species and Related Enzymes in Mitochondrial Respiration During Storage of Harvested Peach Fruits

Peach fruits [Prumus persica （L.） Batsch, cv. Yuhuasanhao] were used as materials to investigate the changes of reactive oxygen species （ROS） and related enzymes in mitochondria respiration during storage and then their influence on senescence of harvested Peach fruits was studied. The results showed that low temperature （5℃） strongly inhibited the reduction of firmness and the increase in respiration rate. During storage at ambient temperature （20℃）, ROS had a cumulative process while malondialdehye （MDA） content continued to increase in associated with enhanced membrane lipid peroxidation. Lipoxygenase （LOX） activity was strongly inhibited under the low temperature condition. The activities of succinic dehydrogenase （SDH）, cytochrome C oxidase （CCO）, and Ca^2＋-ATPase declined to a certain extent at ambient temperature, while they showed higher activities at low temperature, which may be related to lower membrane lipid peroxidation at low temperature. Higher Ca^2＋ content at ambient temperature may be responsible for impairment of mitochondrial function, thus, leading to fruit senescence. The results showed that under low temperature condition, the low accumulation of ROS and the low level of membrane lipid peroxidation could maintain the function of mitochondria that would help to delay the senescence of peach fruits. These suggested a close relationship existed between ROS metabolism and mitochondrial respiration. It can be inferred that the low temperature helps to delay senescence of peach fruits via suppression of ROS and related enzymes, maintain better homeostasis of Ca^2＋ in mitochondria and thus better mitochondrial functions.

Excessive Cu and Zn affecting on distribution of the metals and activities of glycolytic and nitrogen incorporating key enzymes in mycelia of ectomycorrhizal fungi Suillus bovinus

Concentration of copper and zinc in isolated Suillus bovinus mycelia, used nutrient solution and 0.5 mol/L EDTA mycelia washing solution were measured to investigate the distribution of heavy metals in mycelia growth in excess copper or zinc nutrient solution. Treated with zinc, most of added zinc maintained in used solution, and 9.8%/14.6% was in/on mycelia in treatment, and in treatment 2 was 3.9%/8.0% in/on mycelia. In the copper applications, copper stimulated in more than on mycelia, i.e., 25.9%/4.5% in/on mycelia in treatment, and 7%/18.8% in/on mycelia while most of copper retained in used nutrient solution. Certain amount of copper or zinc uptake by mycelia led to pronounced influence on glycolysis and nitrogen incorporating process of Suillus bovinus, while the tested enzymes kept constant in treatment. In crude extracts of copper treatment 2 mycelia, activities of HK, PFK and GS were inhibited and decrease to 63%, 48% and 38% and GIDH increased by 68% of the control, respectively. The behaviors of these tested enzymes toward zinc corresponded in general with that towards copper. The potential protection of Suillus bovinus for its host plant under excess copper or zinc threaten was discussed.

The Study on the Effects of Chinese Herbal Mixtures on Growth,Activity of Post-Ruminal Digestive Enzymes and Serum Antioxidant Status of Beef Cattle

Sixteen Limousin×Luxi crossbred beef cattle [（199.4±8.67） kg] were used to evaluate the effects of Chinese herbal mixtures （CHM） on growth, activities of post-ruminal digestive enzymes and serum antioxidant status of beef cattle. Treatments were control （no CHM supplementation） and three formulations of CHM （CHM-A, CHM-B and CHM-C） that were arranged as completely randomized design. The calves were fed in indoor individual feeding barn with total mixed ration for 90 d. The CHM-A, CHM-B, and CHM-C was incorporated into the diet at the level of 20 g kg-1 DM （dry matter） of concentrate. The cattle were weighted at the beginning, at the end and every 15 d between to determine growth rate. Blood sample was taken every 30 d to analyze activities of total superoxide dismutase （T-SOD） and glutathione peroxidase （GSH-PX） and concentration of malondialdehyde （MDA）. The cattle were slaughtered at the end of experiment and chymus samples from various section of the post-ruminal digestive tract were collected to analyze enzymes activities. All cattle had similar average daily gain, but cattle supplemented with CHM-A tended faster growth （P=0.08） compared to the control and other CHM treatments. Supplementation of CHM did not affect pH of chymus in any section of the post-ruminal digestive tract. Activities of pepsin, trypsin, chymotrypsin, and lipase in chymus samples were enhanced （P0.05） to various extents by supplementation of CHM-A, CHM-B, and CHM-C depending on the enzymes and the site of the digestive tract. Amylase activity was increased only by CHM-A and CHM-C from chymus sample collected at the ileum. Supplementation of CHM-A and CHM-B reduced （P0.05） concentration of MDA, but did not affect GSH-PX activity in the serum of cattle throughout the entire experimental period. Cattle supplemented with CHM-B also had higher （P0.05） T-SOD activity in the serum at day 30 and 60 as compared with the control and the other two CHM treatments. Chinese herbal mixtures supplemented at the levels of 20 g kg-1 DM of concentrate increased post-ruminal digestive enzymes activity and enhanced serum antioxidant status. Inclusion of CHM-A and CHM-B in the diet may be favourable for ruminant production.

Cobalt and manganese stress in the microalga Pavlova viridis (Prymnesiophyceae):Effects on lipid peroxidation and antioxidant enzymes

Pollution of marine environment has become an issue of major concern in recent years. Serious environmental pollution by heavy metals results from their increasing utilization in industrial processes and because most heavy metals are transported into the marine environment and accumulated without decomposition. The aim of the present study is to investigate the effects on growth, pigments, lipid peroxidation, and some antioxidant enzyme activities of marine microalga Pavlova viridis, in response to elevated concentrations of cobalt （Co） and manganese （Mn）, especially with regard to the involvement of antioxidative defences against heavy metal-induced oxidative stress. In response to Co^2＋, lipid peroxidation was enhanced compared to the control, as an indication of the oxidative damage caused by metal concentration assayed in the microalgal cells but not Mn^2＋. Exposure of Pavlova viridis to the two metals caused changes in enzyme activities in a different manner, depending on the metal assayed： after Co^2＋ treatments, total superoxide dismutase （SOD） activity was irregular, although it was not significantly affected by Mn^2＋ exposure. Co^2＋ and Mn^2＋ stimulated the activities of catalase （CAT） and glutathione （GSH）, whereas, glutathione peroxidase （GPX） showed a remarkable increase in activity in response to Co^2＋ treatments and decreased gradually with Mn^2＋ concentration, up to 50 μmol/L, and then rose very rapidly, reaching to about 38.98% at 200 μmol/L Mn^2＋. These results suggest that an activation of some antioxidant enzymes was enhanced, to counteract the oxidative stress induced by the two metals at higher concentration.