Effects of Inoculation with Phosphate Solubilizing Bacteria on the Physiology,Biochemistry,and Expression of Genes Related to the Protective Enzyme System of Fritillaria taipaiensis P.Y.Li

Fritillaria taipaiensis P.Y.Li is a widely used medicinal herb in treating pulmonary diseases.In recent years,its wild resources have become scarce,and the demand for efficient artificial cultivation has significantly increased.This article is the first to apply phosphate solubilizing bacteria isolated from the rhizosphere soil of F.taipaiensis P.Y.Li to the cultivation process of F.taipaiensis P.Y.Li.The aim is to identify suitable reference strains for the artificial cultivation and industrial development of F.taipaiensis P.Y.Li by examining the effects of various phosphate solubilizing bacteria and their combinations on photosynthesis,physiological and biochemical properties,and gene expression related to the protective enzyme system in F.taipaiensis P.Y.Li.The experiment,conducted in pots at room temperature,included a control group(CK)and groups inoculated with inorganic phosphorussolubilizing bacteria:W1(Bacillus cereus),W2(Serratia plymuthica),W12(Bacillus cereus and Serratia plymuthica),and groups inoculated with organophosphorus-solubilizing bacteria:Y1(Bacillus cereus),Y2(Bacillus cereus),Y12(Bacillus cereus and Bacillus cereus),totaling seven groups.Compared to CK,most growth indices in the bacterial addition groups showed significant differences,with W12 achieving the highest values in all indices except the leaf area index.The content of photosynthetic pigments,photosynthetic parameters,and osmoregulatory substances increased variably in each bacterial treatment group.W12 exhibited the highest content of chlorophyll a and soluble protein,while W1 had the highest free proline content.The activities of peroxidase(POD),superoxide dismutase(SOD),and catalase(CAT)in all inoculated groups were higher than in CK,with significant changes in SOD and CAT activities.The malondialdehyde(MDA)content in all inoculated groups was lower than in CK,with Y12 being the lowest,at approximately 30%of CK.Gene expression corresponding to these three enzymes also increased variably,with POD expression in Y2 being the highest at 2.73 times that of CK.SOD and CAT expression in Y12 were the highest,at 1.84 and 4.39 times that of CK,respectively.These results indicate that inoculating phosphate solubilizing bacteria can enhance the growth of F.taipaiensis P.Y.Li,with the mixed inoculation groups W12 and Y12 demonstrating superior effects.This lays a theoretical foundation for selecting bacterial fertilizers in the cultivation process of F.taipaiensis P.Y.Li.

Effect of powdery mildew on interleaf microbial communities and leaf antioxidant enzyme systems

Chinese peony(Paeonia lactiflora Pall.)is both medicinally and aesthetically beneficial.Powdery mildew is a common fungal disease that seriously jeopardizes the value of numerous species,including peonies as a crop.In order to provide a basis for the prevention and treatment of peony powdery mildew,we examined the microbial diversity,the malondialdehyde(MDA)concentrations and antioxidant enzyme activities of peony leaves infected with three levels of powdery mildew to determine any modifications to the leaf's antioxidant enzyme systems and microbial community structure following the onset of disease.The results show that the MDA content rose as the degree of infection became worse.Antioxidant enzyme activity rose and then declined.Following the initiation of powdery mildew,fungal community diversity decreased,whereas there was not any appreciable change in bacterial communities according to microbial diversity sequencing.The relative abundance of more than half of fungal species decreased,with the bacterial genera displaying both abundant and diminished communities with less pronounced alterations in their community structure after the disease spread.Significant different taxa that were critical to the organization of each microbiome were found.Correlation analysis showed that the relative abundance of powdery mildew pathogenic fungal genus Erysiphe was correlated with those of 11 fungal genera and one bacterial genus.Among them,Aureobasidium,Neosetophoma and Sclerostagonospora showed significant positive correlations with Erysiphe and MDA.

Characteristics of heavy metals in soil of lead-zinc mining areas and changes of antioxidant enzyme systems in maize leaf under Pb stress

Pb,Cu,Cd,Zn content of soil in mining areas and abandoned land,flats of the Pijiang River and farmlands were inves-tigated.On this basis of soil heavy metal pollution,the changes of antioxidant enzyme system in maize(Qiandan 88)under different Pb concentrations(0,20,40,60,80,100,150,200,500,1000,2000,3000 mg/L)stress were studied.The results show that the content of Pb,Cu,Cd,and Zn in soil is the highest in mining areas and abandoned land,followed by flats of the Pijiang River>farmlands,and that the variation range of Pb,Cu,Cd in mining areas and abandoned land are 106.40-2564.72,14.83-490.88,22.57-712.77 mg/kg,respectively,which are higher than that of the other land use types.When maize is under stress of 20-500 mg/L Pb concentration,T-SOD activity of maize leaves increase with the increase of Pb concentration and the highest value is 50.21 U/mg prot,but under Pb concentration>1000 mg/L stress,T-SOD activity of maize leaves decrease gradually.The activity of POD decreases with the increases of Pb concentra-tion,and the lowest POD activity of leaves in maize with the value of 93.24 U/mg prot is appeared in Pb 1000 mg/L concentration treatment group.MDA content in leaves of maize increases with the increase of the Pb concentration and the highest value is 101.98 nmol/mg prot,then the content of MDA decreases gradually when the Pb concentration is more than 500 mg/L,which indicates that the membrane lipid peroxidation of maize leaves under high concentration of Pb stress is serious and leads to the cell damage.

Effects of High-Temperature Stress on Photosynthetic Characteristics and Antioxidant Enzyme System of Paeonia ostii

Paeonia ostii is an economically important oil crop,which has been widely cultivated in the middle and lower reaches of the Yangtze River in China in recent years.Although P.ostii is highly adaptable to the environment,the prolonged high summer temperature in this region severely inhibits its growth,which adversely affects seed yield and quality.In this study,P.ostii plants were subjected to 20℃/15℃(day/night)and 40℃/35℃(day/night)temperatures for 15 days.The changes in physiological and biochemical indicators of P.ostii under high-temperature stress were initially investigated.The results showed that with the deepening of leaf etiolation,chlorophyll a and chlorophyll b concentration,carotenoid concentration,Soil Plant Analysis Development(SPAD)values and leaf relative water content decreased significantly,while both relative electrical conductivity(REC)and free proline concentration showed an upward trend.Meanwhile,the continuous accumulation of reactive oxygen species(ROS)in P.ostii plants,led to an increased activity of antioxidant enzymes including superoxide dismutase(SOD),peroxidase(POD),catalase(CAT)and ascorbate peroxidase(APX).Moreover,with the extension of the high-temperature treatment,the anatomical structures of P.ostii were destroyed,resulting in a decreased photochemical efficiency of the photosystem II(PSII)reaction center and photosynthesis was inhibited.Taken together,these results provide reference values for understanding the physiological response of P.ostii to hightemperature stress and establish a foundation for further research on the relevant underlying molecular mechanisms.

Salsola imbricata Forssk.ameliorates acetic acid-induced inflammatory bowel disease by modulating dysregulated antioxidant enzyme system and cytokine signaling pathways in mice

Objective:To explore the protective effect of the crude extract of Salsola imbricata against acetic acid-induced inflammatory bowel disease in mice and its mechanism of action.Methods:Ethanolic crude extract of Salsola imbricata was characterized by HPLC.Salsola imbricata extract at different doses was administered and ulcerative colitis was induced by 200μL,7.5%acetic acid and macroscopic parameters were evaluated to assess the homeostatic condition of intestinal mucosa along with hematological and biochemical assays.The levels of malondialdehyde,glutathione peroxidase 1,superoxide dismutase,and catalase were determined in colon tissues.Proinflammatory cytokines including interleukin(IL)-1β,IL-6,and tumor necrosis factor-α(TNF-α)were quantified by ELISA.The extent of tissue damage was assessed by histological analysis.Results:Phytochemical analysis confirmed the presence of phytochemicals including quercetin,gallic acid,syringic acid,benzoic acid and chlorogenic acid in the crude extract.The crude extract of Salsola imbricata(300 and 500 mg/kg)markedly decreased malondialdehyde and nitric oxide(P<0.01)and increased antioxidant activities of glutathione peroxidase 1(P<0.001)and superoxide dismutase(P<0.001).Moreover,it decreased the levels of IL-1β,IL-6 and TNF-αsignificantly(P<0.001)and reduced the damage to the colon mucosa,promoting tissue healing and regeneration.Conclusions:Salsola imbricata extract restores the colonic epithelial layers by maintaining mucosal homeostasis and cell integrity by modulating antioxidant defense system and inflammatory cytokine signaling in ulcerative colitis mice.

Analytical Expression for the Concentration of Substrate and Product in Immobilized Enzyme System in Biofuel/Biosensor

In this paper, an approximate analytical method to solve the non-linear differential equations in an immobilized enzyme film is presented. Analytical expressions for concentrations of substrate and product have been derived for all values of dimensionless parameter. Dimensionless numbers that can be used to study the effects of enzyme loading, enzymatic gel thickness, and oxidation/ reduction kinetics at the electrode in biosensor/biofuel cell performance were identified. Using the dimensionless numbers identified in this paper, and the plots representing the effects of these dimensionless numbers on concentrations and current in biosensor/biofuel cell are discussed. Analytical results are compared with simulation results and satisfactory agreement is noted.

Angiotensin-converting enzyme 2 alleviates liver fibrosis through the renin-angiotensin system

The present letter to the editor is related to the study titled‘Angiotensin-converting enzyme 2 improves liver fibrosis in mice by regulating autophagy of hepatic stellate cells’.Angiotensin-converting enzyme 2 can alleviate liver fibrosis by regulating autophagy of hepatic stellate cells and affecting the renin-angiotensin system.

Role of renin-angiotensin system/angiotensin converting enzyme-2 mechanism and enhanced COVID-19 susceptibility in type 2 diabetes mellitus

Coronavirus disease 2019(COVID-19)is a disease that caused a global pandemic and is caused by infection of severe acute respiratory syndrome coronavirus 2 virus.It has affected over 768 million people worldwide,resulting in approx-imately 6900000 deaths.High-risk groups,identified by the Centers for Disease Control and Prevention,include individuals with conditions like type 2 diabetes mellitus(T2DM),obesity,chronic lung disease,serious heart conditions,and chronic kidney disease.Research indicates that those with T2DM face a hei-ghtened susceptibility to COVID-19 and increased mortality compared to non-diabetic individuals.Examining the renin-angiotensin system(RAS),a vital regulator of blood pressure and pulmonary stability,reveals the significance of the angiotensin-converting enzyme(ACE)and ACE2 enzymes.ACE converts angiotensin-I to the vasoconstrictor angiotensin-II,while ACE2 counters this by converting angiotensin-II to angiotensin 1-7,a vasodilator.Reduced ACE2 exp-ression,common in diabetes,intensifies RAS activity,contributing to conditions like inflammation and fibrosis.Although ACE inhibitors and angiotensin receptor blockers can be therapeutically beneficial by increasing ACE2 levels,concerns arise regarding the potential elevation of ACE2 receptors on cell membranes,potentially facilitating COVID-19 entry.This review explored the role of the RAS/ACE2 mechanism in amplifying severe acute respiratory syndrome cor-onavirus 2 infection and associated complications in T2DM.Potential treatment strategies,including recombinant human ACE2 therapy,broad-spectrum antiviral drugs,and epigenetic signature detection,are discussed as promising avenues in the battle against this pandemic.

Electro-enzyme coupling systems for selective reduction of CO_(2)

To address the energy crisis and alleviate the rising level of CO_(2)in the atmosphere,various CO_(2)capture and utilization(CCU)technologies have been developed.The use of electro-enzyme coupling systems is a promising strategy for the sustainable production of fuels,chemicals and materials using CO_(2)as the feedstock.In this review,the recent progresses in the development of electro-enzyme coupling systems for the selective reduction of CO_(2)are systematically summarized.We first provide a brief background about the significance and challenges in the direct conversion of CO_(2)into value-added chemicals.Next,we describe the materials and strategies in the design of electrodes,as well as the common enzymes used in the electro-enzyme coupling systems.Then,we focus on the state-of-the-art routes for the electro-enzyme coupling conversion of CO_(2)into a variety of compounds(formate,CO,methanol,C≥2chemicals)by a single enzyme or multienzyme systems.The emerging approaches and materials used for the construction of electro-enzyme coupling systems to enhance the electron transfer efficiency and the catalytic activity/stability are highlighted.The main challenges and perspectives in the integration of enzymatic and electrochemical strategies are also discussed.

Effects of desert plant communities on soil enzyme activities and soil organic carbon in the proluvial fan in the eastern foothills of the Helan Mountain in Ningxia,China

It is of great significance to study the effects of desert plants on soil enzyme activities and soil organic carbon(SOC)for maintaining the stability of the desert ecosystem.In this study,we studied the responses of soil enzyme activities and SOC fractions(particulate organic carbon(POC)and mineral-associated organic carbon(MAOC))to five typical desert plant communities(Convolvulus tragacanthoides,Ephedra rhytidosperma,Stipa breviflora,Stipa tianschanica var.gobica,and Salsola laricifolia communities)in the proluvial fan in the eastern foothills of the Helan Mountain in Ningxia Hui Autonomous Region,China.We recorded the plant community information mainly including the plant coverage and herb and shrub species,and obtained the aboveground biomass and plant species diversity through sample surveys in late July 2023.Soil samples were also collected at depths of 0–10 cm(topsoil)and 10–20 cm(subsoil)to determine the soil physicochemical properties and enzyme activities.The results showed that the plant coverage and aboveground biomass of S.laricifolia community were significantly higher than those of C.tragacanthoides,S.breviflora,and S.tianschanica var.gobica communities(P<0.05).Soil enzyme activities varied among different plant communities.In the topsoil,the enzyme activities of alkaline phosphatase(ALP)andβ-1,4-glucosidas(βG)were significantly higher in E.rhytidosperma and S.tianschanica var.gobica communities than in other plant communities(P<0.05).The topsoil had higher POC and MAOC contents than the subsoil.Specifically,the content of POC in the topsoil was 18.17%–42.73%higher than that in the subsoil.The structural equation model(SEM)indicated that plant species diversity,soil pH,and soil water content(SWC)were the main factors influencing POC and MAOC.The soil pH inhibited the formation of POC and promoted the formation of MAOC.Conversely,SWC stimulated POC production and hindered MAOC formation.Our study aimed to gain insight into the effects of desert plant communities on soil enzyme activities and SOC fractions,as well as the drivers of SOC fractions in the proluvial fan in the eastern foothills of the Helan Mountain and other desert ecosystems.

Ellagic Acid Enhances Antioxidant System Activity and Maintains the Quality of Strawberry Fruit during Storage

Ellagic acid(EA)is a natural antioxidant,widely present in a lot of forms’soft fruits,nuts,and other plant tissues,and helpful for promoting human health;however,its protective effect on postharvest fruit and improving the quality index of postharvest fruit have rarely been studied.In this experiment,the strawberries were soaked in 0,100,200,300,400,and 500 mg L^(−1) EA,respectively,and the influential EA on fruit quality and the antioxidant system of strawberries were studied.Compared with the control,EA treatment can reduce the browning degree and rotting rate of strawberry fruit during storage and augment the soluble solid content(SSC).EA treatment can also increase the content of related stuff and enzyme activity in antioxidant systems;the gene expression level of polyphenol oxidase(PPO)in strawberries treated with EA was always down-regulated,correspondingly,the expression of other antioxidant enzyme genes was enhanced.Among the strawberry fruits treated with EA of different concentrations,300 mg L^(−1) EA had the best effect in the process of strawberry preservation.The results suggested that the proper concentration of exogenous EA at 300 mg L−1 could maintain strawberries’quality and enhance the antioxidant system by improving the activities of antioxidative enzymes and the ascorbateglutathione(AsA-GSH)cycle during storage.

Extracellular vesicles and angiotensin-converting enzyme 2 in COVID-19 disease

Extracellular vesicles(EVs)are membranous vesicular structures released from almost all eukaryotic cell types under different physiological or pathological conditions.Growing evidence demonstrates that EVs can serve as mediators of intercellular communication between donor and recipient cells or microorganism-infected and noninfected cells.Coronavirus disease 2019(COVID-19)disease is caused by infection of the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)of host cells in the respiratory system and various extra-pulmonary tissue/organs,resulting in complications of multiple organ systems.As the cell surface receptor,angiotensin-converting enzyme 2(ACE2)mediates cellular entry of SARS-CoV-2 into the host cells in patients with COVID-19.Recent studies have found that ACE2 can be released with EVs,which have been shown to interfere with the entry of the virus into host cells and thus may be involved in COVID-19 pathophysiology.In addition,ACE2,neprilysin(NEP),and thimet oligopeptidase(TOP)are the key enzymes that regulate angiotensin metabolism by converting angiotensin II or angiotensin I to angiotensin 1-7,the latter of which has protective effects in counterbalancing the harmful effects of angiotensin II in COVID-19 disease.This review summarizes the recent research progress regarding EV-associated ACE2,NEP,and TOP and the perspectives of their potential involvement in the pathophysiology of COVID-19 disease.

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.

Study on Antioxidant Enzyme Activity and Physiology and Biochemistry of Solanum nigrum L. under Glyphosate Stress

[Objectives] This study was conducted to investigate the scientific prevention and control of Solanum nigrum L. [Methods] Through experiments on S. nigrum from different sources, it was found that glyphosate stress had significant effects on antioxidant enzyme activity and oxidative damage of sensitive S. nigrum plants. [Results] Sensitive S. nigrum showed oxidative damage under glyphosate stress, while resistant S. nigrum responded to adversity damage by improving its antioxidant enzyme activity. The experimental results showed that the antioxidant enzymes and reduced glutathione of S. nigrum had certain metabolic detoxification effects under glyphosate stress. [Conclusions] This study provides a theoretical basis for scientific prevention and control of S. nigrum , and has a certain reference value for revealing the glyphosate resistance mechanism of S. nigrum .

Microplastic Can Decrease Enzyme Activities and Microbes in Soil

An in vitro study was conducted to investigate the impacts of microplastics on enzyme activities and soil bacteria. The study included four different treatments of microplastics including a control. Different levels of microplastics were applied to the soil ranging from 0% to 5%, to assess the impacts of microplastics on soil enzymes and subsequent soil bacteria. After 30 days of incubation, the soil samples were collected and growth parameters of bacteria were assessed. Activities of β-glucosidase, urease and dehydrogenase enzymes were also determined. Our results showed that the presence of microplastics in the soil significantly reduced bacterial population together with bacterial strains. The activities of β-glucosidase, urease and dehydrogenase enzymes were reduced significantly to approximately 32%, 40% and 50% in microplastics treated soils respectively. Concentration of microplastic has a role to play towards this direction;the higher the concentration of microplastic the greater is the impact on enzymes and soil bacteria. The present study on the microbial soil health vis-à-vis microplastic application indicates that the material can have negative effect on the soil bacterial population of and thus ultimately may jeopardize soil health and crop production.

The Therapeutic Effect of Biling Weitong Granules Combined with Oryz-Aspergillus Enzyme and Pancreatin Tablet on Reflux Esophagitis with Functional Dyspepsia

Objective:To investigate the therapeutic effect of Biling Weitong Granules combined with oryz-aspergillus enzyme and pancreatin tablets on patients with reflux esophagitis with functional dyspepsia.Methods:Sixty patients diagnosed with reflux esophagitis with functional dyspepsia who were admitted to the Affiliated Hospital of Hebei University between June 2020 and June 2023 were selected and divided into two groups:the control group and the observation group,each consisting of 30 cases.The control group received oryz-aspergillus enzyme and pancreatin tablets only,while the observation group received Biling Weitong Granules in addition to the tablets.The clinical efficacy,Chinese medicine syndrome points,esophageal kinetic indexes,gastrointestinal hormone levels,and therapeutic safety of both groups were evaluated.Results:The total efficiency of the observation group reached 93.33%,significantly higher than the 73.33%of the control group(P<0.05).After treatment,patients in the observation group exhibited significantly lower scores for Chinese medicine symptoms such as early satiety,belching,abdominal distension,abdominal pain,and loss of appetite compared to the control group(P<0.05).Furthermore,the observation group showed significantly higher upper esophageal sphincter pressure,lower esophageal sphincter pressure,and distal esophageal contraction scores compared to the control group(P<0.05).Additionally,levels of gastric motility hormone,vasoactive intestinal peptide,and gastrin were significantly higher in the observation group compared to the control group(P<0.05).Throughout the treatment period,there was no significant difference in the incidence of adverse reactions between the two groups,indicating comparable safety of the two treatment modalities(P>0.05).Conclusion:The combination of Biling Weitong Granules with oryz-aspergillus enzyme and pancreatin tablets demonstrates significant efficacy in the treatment of reflux esophagitis with functional dyspepsia,with a better safety profile.This finding warrants further clinical promotion.

Analytical Solutions of System of Non-Linear Differential Equations in the Single-Enzyme, Single-Substrate Reaction with Non-Mechanism-Based Enzyme Inactivation

A closed form of an analytical expression of concentration in the single-enzyme, single-substrate system for the full range of enzyme activities has been derived. The time dependent analytical solution for substrate, enzyme-substrate complex and product concentrations are presented by solving system of non-linear differential equation. We employ He’s Homotopy perturbation method to solve the coupled non-linear differential equations containing a non-linear term related to basic enzymatic reaction. The time dependent simple analytical expressions for substrate, enzyme-substrate and free enzyme concentrations have been derived in terms of dimensionless reaction diffusion parameters ε, λ1, λ2 and λ3 using perturbation method. The numerical solution of the problem is also reported using SCILAB software program. The analytical results are compared with our numerical results. An excellent agreement with simulation data is noted. The obtained results are valid for the whole solution domain.

Effects of excessive nitrogen fertilizer and soil moisture deficiency on antioxidant enzyme system and osmotic adjustment in tomato seedlings

Long-term excessive application of nitrogen fertilizer induces secondary salinization of soil,which results in inhibiting plant growth.In addition,soil moisture deficiency also affects plant growth.To investigate the effects of excessive nitrogen fertilizer and soil moisture deficiency on the antioxidant enzyme system,plant water relations analyzed through pressure-volume(P-V)curve,and photosynthetic light response parameters in tomato(Solanum lycopersicum L.Myoko)seedlings,an indoor experiment of about 50 d was conducted using two irrigation water amounts based on field capacity(soil moisture deficiency:50%-80%;adequate water:70%-80%),two nitrogen fertilizer rates(moderate nitrogen;excessive nitrogen fertilizer:0.585 g/pot)and two types of irrigation water(tap water and microbial diluent).The results showed that excessive nitrogen fertilizer(N)and soil moisture deficiency(W)reduced the biomass of tomato seedlings.In comparison to CK(combination of adequate water and tap water quality),microbial dilution(EM)increased plant biomass by 5.2%.Also,the nitrogen application increased chlorophyll relative contents(SPAD).The maximum net photosynthetic rate(Pc)decreased with nitrogen application and increased with EM application and irrigation amount.Excessive nitrogen application increased the plant nitrate reductase activity(NR).The plant NR in the N treatment showed a 13.0%increase compared to CK,and the plant NR in the treatment of nitrogen application with water deficiency(WN)increased 34.0%compared to water deficiency(W).After applying excessive nitrogen,N,EM-N,WN,EM-WN respectively increased the plant nitrate reductase activity by 13.0%,22.9%,34.0%,and 28.6%,compared with the corresponding treatment with moderate nitrogen(i.e.,CK,EM,W and EM-W).In addition,the activities of antioxidant enzymes[superoxide dismutase(SOD),peroxidase(POD)and catalase(CAT)]in four treatments of nitrogen application(N,EM-N,WN,EM-WN)also increased significantly.Both soil moisture and nitrogen fertilizer significantly affect the parameters of osmotic adjustment,which is manifested in the reduction of osmotic potential(π_(FT)),and the increase in the osmotic concentration(C_(osm))and concentration difference(ΔC_(osm)).But the decrease in the relative water content of apoplast(ζ_(ap))indicated that water deficiency and excessive nitrogen reduced the water absorption and water retention capacity of tomatoes to a certain extent.In conclusion,excessive nitrogen application and soil moisture deficiency inhibit plant growth significantly in this experiment.Meanwhile,microbial dilution can alleviate excessive nitrogen fertilizer and water stress to some extent,but the effect was not significant.

Effects of broccoli extract and various essential oils on intestinal and faecal microflora and on xenobiotic enzymes and the antioxidant system of piglets

Objective: Since the ban of antibiotics as growth promoting feed additives in the EU in 2006 research in alternatives has gained importance. Phytogenic feed additives represent a heterogenous class of different plant derived substances that are discussed to improve the health of farm animals by direct and indirect antioxidant effects and by influencing microbial eubiosis in the gastrointestinal tract. Consequently our study aimed to investigate the influence of broccoli extract and the essential oils of tur- meric, oregano, thyme and rosemary, as selected individual additives, on intestinal and faecal microflora, on xenobiotic enzymes, and on the antioxidant system of piglets. Methods: 48 four weeks old male weaned piglets were assigned to 6 groups of 8. The piglets were housed individually in stainless steel pens with slatted floor. The control group (Con) was fed a diet without an additive for 4 weeks. The diet of group BE contained 0.15 g/kg sulforaphane in form of a broccoli extract. 535, 282, 373 and 476 mg/kg of the essential oils of turmeric (Cuo), oregano (Oo), thyme (To) and rosemary (Ro) were added to the diets of the remaining 4 groups to stan-dardise supplementation to 150 mg/kg of the oils’ key terpene compounds ar-turmerone, carvacrol, thymol and 1,8-cineole. The composition of bacterial microflora was examined by cultivating samples of jejeunal and colonic mucosa and of faeces under specific conditions. The mRNA expression of xenobiotic and antioxidant enzymes was determined by reversing transcrip- tase real time detection PCR (RT-PCR). Total antioxidant status was assayed using the Trolox Equivalent Antioxidant Capacity (TEAC), and lipid peroxidation was determined by measuring thiobarbioturic acid reactive substances (TBA- RS). Results: Compared to Con piglets all additives positively influenced weight gain and feed conversion in week 1. Over the whole trial period no significant differences in performance parameters existed between the experimental groups. Compared to group Con performance of Ro piglets was, however, slightly impaired. Com- pared to Con piglets Cuo, Oo and To increased the ratio of Lactobacilli:E. coli attached to the jejunal mucosa, whereas BE and Ro impaired this ratio slightly. In contrast in colonic mucosa Ro improved Lactobacilli:E. coli ratio. In faecal samples an improvement of Lactobacilli:E. coli ratio could be analysed for To and Ro. Ro was the only additive that reduced the incidence rate of piglets tested positive for enterotoxic E. coli (ETEC). All additives significantly increased jejunal TEAC and reduced TBA-RS. In the liver BE, Cuo, Oo and To increased TEAC in tendency and Ro significantly. Liver TBA-RS were slightly reduced by all additives compared to Con piglets. Whereas the influence of BE, To and Ro on jejunal TEAC mainly was derived from the induction of xenobiotic and antioxidant enzymes (indirect antioxidant effects), Cuo and Oo influenced TEAC by direct antioxidant effects. Discussion and Conclusions: Our results have shown: That within the labiatae oils Oo and To have the potential to improve performance slightly. That phytogenic substances have a small but not sig- nificant influence on intestinal microflora. That phytogenic feed additives up-regulate the anti- oxidant system of piglets either by direct or by indirect antioxidant effects and that they may thereby improve health status. That within the labiatae oils Oo has a high direct antioxidant potential whereas Ro potently induces xenobiotic and antioxidant enzymes. That broccoli extract is an attractive new phytogenic additive, improving antioxidant status by indirect antioxidant effects. That defined combinations of selected phytogenic substances may produce additive effects. That health promoting effects of phytogenic additives in the future should be studied systematically under the challenge with pathogenic microorganisms or food derived to-xins.

Enzyme electrophoresis method in analysis of active components of haemostasis system

The novel modifications of substrate-containing sodium dodecyl sulfate-polyacrylamide gel electrophoresis that can be used for the detection of proteases and its activators are reported. The protease/activator samples were separated on a protein substrate-SDS-polyacrylamide gel. To detect plasminogen activators fibrinogen and Glu-plasminogen were incorporated into the SDS-PAG followed by 1 h incubation at 37?C in thrombin solution (1 NIH/ml). After electrophoresis the gel was stained according to the standard protocol. To detect fibrin-unspecific plasminogen activators from snake venom incubation in thrombin solution was substituted for 12 h incubation in 50 mM Tris-HCl (pH 7.4). To detect fibrinogen-degrading enzymes fibrinogen-containing gel was used. Activity of protease/activator was visualized in the gel as clear bands against the dark background. These new techniques offer several advantages including determination of the quantity and activity of t-PA and urokinase, however cannot be recommended for precise quantification of activators;the total procedure is quite quick and simple;method is convenient tool for detection of novel protein-protein interactions in haemostasis system;the sensitivity of the method is ≤0.01 IU per track.