RESEARCH ON THE HEREDITY AND VARIATION OF THE PERUXIDASE ISOENZYME OF NEEDLES OF PINUS KORAIENSIS

Through the polyacrylamide gel clectrophoresis analysis of thirty-two needle samples of ninty-years old pinus koraiensis in Harbin, seven typies of zone and distribution of the zone frequency, and some objective fact of heredity and variation were determined. These informations can be used as reference materials in the research of heredity and variation of pinus koraiensis.

The combination of creatine kinase-myocardial band isoenzyme and point-of-care cardiac troponin/contemporary cardiac troponin for the early diagnosis of acute myocardial infarction

BACKGROUND:The early diagnosis of acute myocardial infarction(AMI)remains challenging,especially for institutions without the high-sensitive cardiac troponin(hs-c Tn)assay.Herein,we aim to assess the value of creatine kinase-myocardial band isoenzyme(CK-MB)combined with different cardiac troponin(c Tn)assays in AMI diagnosis.METHODS:This multicenter,observational study included 3,706 patients with acute chest pain from September 1,2015,to September 30,2017.We classified the participants into three groups according to the c Tn assays:the point-of-care c Tn(POC-c Tn)group,the contemporary c Tn(c-c Tn)group,and hs-c Tn group.The diagnostic value was quantified using sensitivity and the area under the curve(AUC).RESULTS:Compared to the single POC-c Tn/c-c Tn assays,combining CK-MB and POC-c Tn/c-c Tn increased the diagnostic sensitivity of AMI(56.1%vs.63.9%,P<0.001;82.7%vs.84.3%,P=0.025).In contrast,combining CK-MB and hs-c Tn did not change the sensitivity compared with hs-c Tn alone(95.0%vs.95.0%,P>0.999).In the subgroup analysis,the sensitivity of combining CKMB and c-c Tn increased with time from symptom onset<6 h compared with c-c Tn alone(72.8%vs.75.0%,P=0.046),while the sensitivity did not increase with time from symptom onset>6 h(97.5%vs.98.3%,P=0.317).The AUC of the combination of CK-MB and POC-c Tn significantly increased compared to the single POC-c Tn assay(0.776 vs.0.750,P=0.002).The AUC of the combined CKMB and c-c Tn/hs-c Tn assays did not significantly decrease compared with that of the single c-c Tn/hs-c Tn assays within 6 h.CONCLUSIONS:The combination of CK-MB and POC-c Tn or c-c Tn may be valuable for the early diagnosis of AMI,especially when hs-c Tn is not available.

Repellent Activity of Extracts of Wild Rice Species against Panonychus citri and Aphis citricola in Associated with Esterase Isoenzyme in Insests

Six species of wild rice with different ecophenotypes including Oryza grandiglumis （E6-1, E6-3 / 6-4）, O. minuta （E13-9, E13-13）, O. officinalis（E15-8, E15-13）, O. punctata （E16-1, E16-3, E16-13）, O. granulata（E7-4）, and O. latifolia（101392, E9-1, E9-10） were extracted with methnol and the repellent activity of the extracts against the two insects Aphis citricola and Panonychus citri were studied. The extracts of O. officinalis E15-8 showed higher repellent rate to the two insects than those of the other species. The repellent rates of the extracts of E15-8 to P. citriand A. citricola were 83.26% and 87.86% at 5×10^4 μg/mL in 24 h and 87.95% and 82.43% in 48 h, respectively. The extracts of O. officinalis E15-8 had the effect of inhibition to the esterase of the two insects.

Adenosine deaminase isoenzymes estimation - as a diagnostic tool for tuberculous pleural effusions

Objective:To assess the efficacy of ADA isoenzyme estimation over that of total ADA level in pleural fluid and serum as a more efficient diagnostic indicator in tuberculous pleural effusions in high prevalence country like India.Methods:The efficacy was analysed in total thirty four patients of pleural effusions.Total ADA was estimated by Guitsi and Galanti Calorimetric method and ADA isoenzymes with and without EHNA[Erythro-9-（2- hydroxy-3-nonyl） adenine]a potent ADA1 inhibitor using the same method.Results:The results demonstrated a statistically significant values of ADA2 in serum（P【0.001）,pleural fluid（P = 0.000） and significant value for the ratio of pleural fluid ADA2/serum ADA2（P【0.001） and pleural fluid ADA/ADA(2（P【0. 005）.The sensitivity and specificity values of pleural fluid ADA|2 is 81.8%and 91.6%（cut off value 60 IU/L for Tuberculous effusions）,serum ADA2 95.4%and 66%（cut off value 70 IU/L for tuberculous effusions）. ADA2 is an isoenzyme,which is significantly raised in tuberculous pleural effusions both in the serum and pleural fluid.Conclusion:Estimation of ADA isoenzymes is redundant as a diagnostic aid over total ADA estimation in view of the limited improvements both in specificity and sensitivity patterns and also in term of cost-benefit ratio.

Variation of peroxidase isoenzyme and biofilm of Phanerochaete chrysosporium in continuous membrane bioreactor for Reactive Brilliant Red X3-B treatment

The influence of a Reactive Brilliant Red X-3B （RBR X-3B） dye on the peroxidase isoenzyme ofPhanerochaete chrysosporium was determined, and the biofilm structure in a white rot fungal continuous membrane bioreactor （MBR） was also investigated by scanning electron microscope （SEM）. The variation of peroxidase isoenzyme and the decolorization rate in the continuous MBR were evaluated. The results showed that the 100 mg/L RBR X-3B could stimulate the production of the peroxidase isoenzyme in the shaking-flask culture. In addition, two new peroxidase isoenzyme bands with relative mobility （Rf） value of 0.27 and 0.28 appeared, but the activity was lower than the blank control of 11 d. In the continuous MBR, the system worked stably during the first 60 d, the main peroxidase isoenzyme bands existed and three new bands with Rf value of 0.10, 0.27, and 0.28 appeared. Meanwhile, the biofilm grew well and the average decolorization rate could reach 90.6%. But the bands of peroxidase isoenzyme decreased rapidly at day 65, only two bands with Rf value 0.24 and 0.26 existed, and the decolorization rate decreased to 78.3%. Therefore, 5 bottles of P. chrysosporium mycelial pellet were added into the MBR, and then the activity of the peroxidase isoenzyme and the decolorization rate had a slight recovery. Finally, the decolorization rate finally decreased to 75.2%. These results contribute to a comprehensive understanding of the variation of peroxidase isoenzyme and biofilm in continuous MBR by white rot fungi.

Expression and kinetic changes of alkaline phosphatase and its isoenzymes in experimental rat hepatoma

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Purification, Immobilization and Characterization of Lipase Isoenzyme from <i>Aspergillus niger</i>with C8 Magnetic Particles

The purification of a lipase isoenzyme from an Aspergillus niger lipase A is reported in this manuscript. Purification was carried out in a simple adsorption step, in which the lipase was offered at low ionic strength to the commercially available C8 modified magnetic particles, MaKProt C8. When the isoenzyme was desorbed with a 0.2% solution of Triton X-100, the SDS-PAGE gel showed a single pure band with a molecular weight of 35 KDa. The purified fraction showed 66.75-fold purification compared with the crude extract. The pure fraction was characterized along with the crude extract and the lipase adsorbed on the MaKProt C8. The purified and the adsorbed lipase showed better activity for the tested substrates (p-nitrophenyl acetate, decanoate, myristate and palmitate) than the crude extract, the preferred substrates being myristate (26.7 μmol·min-1·mg-1) and decanoate (17.4 μmol·min-1·mg-1), respectively. The temperature and pH profiles showed no change for the three enzymes, the optimum temperature being 37°C and the best pH 7.0.

AN ENZYMOCYTOCHEMICAL STUDY OF ALKALINE PHOSPHATASE ISOENZYMES IN GASTRIC CANCER－LIGHT AND ELECTRON

By using light and electron microscopic cytochemical technique, the activities and distributions of AP isoenzymes in gastric cancers and benign gastric diseases were examined.The results showed: Nagao, Regan and Kasahara isoenzymes, which were not expressed in normal gastric mucosae and non-malignant lesions, might be considered as tumor markers of gastric cancer;The epithelium of intestinal metaplasia exhibited intestinal-type AP only,which was one of the markers of well-differentiated intestinal metaplasia.In the view-point of the gene expression of AP isoenzymes, two mutation hypothesis and recessive-gene mutation hypothesis might be fit for gastric cancer.

UV-B irradiation and allelopathy by Sargassum thunbergii affects the activities of antioxidant enzymes and their isoenzymes in Corallina pilulifera

Intertidal macroalgae can cope with the dual effects of UV-B irradiation and allelopathy.To study the impacts of the two stressors,we co-cultured Corallina pilulifera with Sargassum thunbergii in 1:1 and 1:10 ratios under different doses of UV-B radiation.The response of the antioxidant defense system,focusing on activities of superoxide dismutase(SOD),ascorbate peroxidase(APX),peroxidase(POX)and glutathione reductase(GR),was monitored.In addition,isoenzyme patterns were analyzed using nondenaturing poly acry lamide gel electrophoresis.The results show that the activities of SOD,APX,and GR were all significantly affected by both UV-B radiation and allelopathy,and the effect of their interaction was significant.However,POX activity was only influenced by UV-B radiation.The enzymatic assay revealed four distinct bands of SOD.The SODⅢband weakened significantly when the co-cultures were exposed to extremely high dosage of UV-B irradiation under both co-culturing ratios of 1:1 and 1:10.When the coculturing ratio was 1:10,both POXⅡand APXⅡenzy me activities increased with different UV-B doses.GR activity was at its greatest when the co-culture ratio was 1:10 and exposure was to the higher UV-B doses.The activities of GRⅢand GRⅣwere elevated under all UV treatments whereas the activities of GRI and GRII were reduced under the lower UV-B treatments but were elevated under the higher UV-B treatments.However,lipid peroxidation,as indicated by the thiobarbituric acid-reacting substance(TBARS)assay,increased significantly under the dual stressors.Our data sugge st that allelopathy and UV-B radiation stre ss can each affect the antioxidant enzyme activities of C.pilulifera.Critically,the adverse effects of UV-B on C.pilulifera were intensified by the compounding effects of allelopathy.

Effect of Heavy Metal Pollution on Cucumber POD Isoenzyme

The Experiment was carried out to study heavy metal pollution effect on cucumber POD isozyme in our experiment.The results showed that:the activities of POD in cucumber seeds increased in low concentration treatment,and decreased in high concentration treatment after being treated with heavy metal.The critical band patterns of POD isoenzyme was changed by heavy metal pollution.

Purification of Active Peroxidase Isoenzymes and Their Responses to Nitrogen Fertilization and Rotation of Biomass Sorghum

Peroxidases (EC 1.11.1.7) participate in lignin biosynthesis. But peroxidation is not a tool for assaying lignocellulose metabolism because the active cannot yet be separated from the inactive peroxidases. A biochemical tool for assaying plant cell wall responses to agronomic practices is needed in the lignocellulosic feedstock renewable energy industry. Peroxidase of biomass sorghum was purified to 9 - 13 charge isomers by free solution IEF (Rotofor) technique. Free solution IEF was more effective than chromatographic purification of active peroxidase isoenzymes. Native PAGE separated each charge isomer to three anionic and three cationic isoenzymes. Hydrogen peroxide and o-dianisidine assays showed that only 20% - 30% of the isoenzymes displayed normal Michaelis-Menten kinetics. Sorghum planted without nitrogen fertilization induced the hydrogen peroxide noncompetitive inhibition of peroxidase, but 280 kg·ha–1 nitrogen fertilization and 100% sorghum mineral residue return to the soil tripled the concentration of active peroxidase and relieved the inhibition with concomitant increases of 350 kg lignin and 3532 kg·cellulose·ha–1. Nitrogen fertilization without crop rotation induced hydrogen peroxide inhibition of the peroxidase, but nitrogen fertilization and 25% sorghum rotation changed the PI of the active peroxidase from neutral to mildly acidic and relieved the inhibition with concomitant enormous increases of 690 kg lignin and 7151 kg·cellulose·ha–1. Hydrogen peroxide inhibition kinetics is consistent with the known peroxidase-substrate intermediate dead-end complex formation. Lignocellulosic yield was greatest under the agronomic management that combined 280 kg·ha–1 nitrogen fertilizer with 25% sorghum residue, which resulted in a shift of pI value of the active peroxidase due to a reduction in the Km value of the peroxidase. Therefore, up to 75% of sorghum biomass rather than only 50% can be harvested for conversion to bioenergy products.

Lactate Dehydrogenase Isoenzymes in Mammalian Cells Exposed to Isophenphos

The cytotoalcity of isophenphos, an organophosphorus insecticide that has the potential to cause delayed polyneuropathy, was evaluated in GBK and V79 ce1ls. A 72 h time course following isophenphos exposure indicated a dose-dependent growth inhibition as dctermined by cell counts. The administration of isophenphos (20g/ml) to GBK cells cultured at high densities indicated a decrease in the activities of LDH isocnzymes. Analysis of V79 cells revealed a decrease of LDH3, the only LDH isocnzyme detected in these cells.

Genetic variability of the Saudi Arabian Uromastyx aegyptia microlepis using protein and isoenzymes electrophoreses

Electrophoresis for SDS-proteins and isoenzymes were conducted to investigate the genetic variations within the agamid lizard Uromastyx aegyptia microlepis inhabiting the desert of Saudi Arabia. Samples were collected from four localities: a) Ushayrah near the town of Taif, b) Al Gwaih near Riyadh, c) Nairyah near Dammam and d) Mouileh near Tabok. A range of 7 to 14 protein bands were recorded in the patterns of the studied samples as measured by SDS-polyacrylamide gel electrophoresis. Among them, only one fraction was recorded in all samples as a common protein band. Six arbitrary chosen enzymes were examined by native-polyacrylamide gel electrophoresis. They were α and β esterase (Est), acid phosphatase (Acph), Alcohol dehydrogenase (Adh), Aldehyde oxidase (Ao) and peroxidase (Px). Seventeen heterogeneous alleles have been recorded;seven of them were fixed in all populations and 10 were polymorphic. Nearly all recorded alleles were monomeric in all samples. α-Est2, β-Est2, Acph2 and Px1 were restricted to Tabok samples and were not recorded in other localities. β-Est3, Acph3, Adh1, Adh1and Px2 were not recorded in Taif samples and the latter one was not recorded in the Dammam samples. The similarity coefficient that has been calculated according to the number of sharing bands indicated the clustering of Tabok and Dammam populations together. The constructed tree based on the sharing protein bands and isozyme alleles revealed similar results regarding the kinship of both populations. The present results indicated that the populations of this subspecies exhibits high genetic variability among its populations.

Tissue-Specific Isoenzyme Variations in Tilapia Fish, <i>Oreochromis niloticus</i>

Native polyacrylamide gel electrophoresis have been used to analyze malate dehydrogenase (MDH), acid phosphatase (Acph) and peroxidase (Px) isoenzymes in different tissues (liver, kidney, muscle and heart) of the tilapia fish, Oreo- chromis niloticus in order to study the tissue specificity of these isoenzymes. Three, two and one fractions have been recorded respectively for the three isoenzymes in different studied tissues. The MDH-1 and MDH-2 have been ex- pressed only in muscle and heart while MDH-3 has been expressed in all studied tissues. The percentage amount of MDH in general varied significantly between muscle and different studied tissues. With respect to acid phosphatase, the percentage amount of the total enzyme showed significant difference between liver and muscle and that this variation may be due to higher gene activity in liver. Peroxidase isoenzyme was recorded in liver and heart only with significant increase in liver. The kidney was the least among the studied tissues in showing gene expression for the studied isoenzymes and therefore, liver, heart and muscle tissues are better applicable in studying the isoenzymatic profiles for fish physiology and systematics.

Relationship Between Serum Creatine Kinase Isoenzyme MM Subbands and the Gradation of Coronary Stenosis in Patients with Unstable Angina Pectoris

Objective To observe the relationship between serum creatine kinase isoenzyme MM sub-bands (CKMM3/MM1 ratio) and the gradation of coronary stenosis and provide a simple, reliable, and economical method for identifying high-risk unstable angina pectoris (UAP). Mehtods Blood samples were drawn at different time after onset of chest pain in 21 patients with UAP and only once in 20 each volunteers for control. CKMM3/MM1 ratio was detected by nonserial buffer agarose gel electrophoresis. CKMB and CK were observed by velocity method. An emergent coronary arteriography was performed as soon as patients were admitted into hospital. Results Patients with UAP were divided into two subgroups: patients with elevated serum enzyme [P( + )] and patients with normal serum enzyme [P( - ) ] according to CKMM3/MM1 ratio < 0. 5. Patients with UAP( + ) had higher serum CKMM3/MM1 ratios from 0.5 to 12hrs and serum CKMB from 2 to 12 hrs than those with UAP( - ) and control ( P < 0. 05) . Serum enzyme concentrations of patients with UAP whose coronary lumen had 90% or more than 90% stenosis were significantly higher than those whose coronary lumen had less than 90% stenosis ( P <0. 01) . Any CK-MM3/MM1 ratio was less than 1. 0 and CK within the normal range in patients with UAP( + ) group. Conclusions CKMM3/MM1 ratios in patients with UAP can reflect severity of myocardial ischemia. SerumCKMM3/MM1 ratio provides a simple, reliable, and economical method for identifying high-risk UAP.

Faecal pyruvate kinase isoenzyme type M2 for colorectal cancer screening:A meta-analysis

AIM:To present a critical discussion of the efficacy of the faecal pyruvate kinase isoenzyme type M2(faecal M2-PK) test for colorectal cancer(CRC) screening based on the currently available studies.METHODS:A literature search in PubMed and Embase was conducted using the following search terms:fecal Tumor M2-PK,faecal Tumour M2-PK,fecal M2-PK,faecal M2-PK,fecal pyruvate kinase,faecal pyruvate kinase,pyruvate kinase stool and M2-PK stool.RESULTS:Stool samples from 704 patients with CRC and from 11 412 healthy subjects have been investigated for faecal M2-PK concentrations in seventeen independent studies.The mean faecal M2-PK sensitivity was 80.3%;the specificity was 95.2%.Four studies compared faecal M2-PK head-to-head with guaiacbased faecal occult blood test(gFOBT).Faecal M2PK demonstrated a sensitivity of 81.1%,whereas the gFOBT detected only 36.9% of the CRCs.Eight independent studies investigated the sensitivity of faecal M2-PK for adenoma(n = 554),with the following sensitivities:adenoma < 1 cm in diameter:25%;adenoma > 1 cm:44%;adenoma of unspecified diameter:51%.In a direct comparison with gFOBT of adenoma > 1 cm in diameter,47% tested positive with the faecal M2-PK test,whereas the gFOBT detected only 27%.CONCLUSION:We recommend faecal M2-PK as a routine test for CRC screening.Faecal M2-PK closes a gap in clinical practice because it detects bleeding and nonbleeding tumors and adenoma with high sensitivity and specificity.

Isoenzymes and soluble protein in Arthrospira from alka-line lakes in Erdos Plateau, China and in exotic species

The authors compared isoenzymes of five enzymes and soluble protein in Arthrospira platensis (A3) and A. erdosensis (A4) from alkaline lakes in Erdos Plateau, Nei Monggo (Inner Mongalia), China and exotic species of A. platensis (A1) from Chad and A. maxima (A2) from Mexico by means of polyacrylamide gel electrophoresis. The results showed that the isoenzymes of EST, POD and soluble protein were polymorphic. Monomorphism and polymorphism were found in isoenzymes of AMY, CAT and SOD, and monomorphism was found only in the introduced species. The isoenzymes and soluble protein of the local species are all polymorphic. The number of bands in these species were in the order of A3>A4>A1>A2. A2 is the most primary, A1 from Chad Lake is relatively primary, A3 and A4 are ad- vanced species in evolution. Cluster analysis showed that the relation between the two introduced species are the closest to each other, and so too are the two local ones.

Tear Malate Dehydrogenase,Lactate Dehydrogenase and Their Isoenzymes in Normal Chinese Subjects and Patients of Ocular Surface Disorders

Purose:To determine levels of malate dehydrogenase(MDH),lactate dehydroge-nase(LDH)and their isoenzymes in tears of normal Chinese subjects and patients with ocular surface disorders.Methods:The age range of normal subjects was10-88,with136mal and 128fe-male subjects.123patients suffered from ocular surface disorders.Tears were col-lected from lower fornix on Xinghua filter disc(0.1mm thick,5mm in diameter).The values of tearMDHand LDHwere determined by MONARCH-2000Ana-lyzer(U.S.A)Their isoenzymes were separated by acetate cellulose elec-trophoresis and were determined by Model CDS-200light densitometer.Results:The normal values of tear LDH and MDH were 45.51+23.00-81.35+37.84umol·s^-1/Land11.00+5.33-19.50+9.17umol·s^-1/Lrespectively,dis-regarding sex or eye distriction(P>0.05).The values of tear LDHandMDH in the group aged10-19were significantly lower than in another groups(P<0.05),95%normal ranges of tearMDHaged below19and above20were3.63-19.90umol·s^-1/L.THe MDH isoenzymes comprised MDHs and MDHm,the former accounting for80.0-89.1%.The LDH isoenzymes comprised 5varieties.of which the ratioH/Mof subunit H tosubunit Mwas0.196+0.02.Levels of tear LDH,MDHand their isoenzymes in different diseases were various.Conclusions;Tear LDH/MDHratio reflected sensitively the matabolism of corneae and conjunetival epithelium.The changes in LDH isoenzymes were hel-ful to the differential diagnosis of external eye diseases,and the increase of MDHm reflected sensitively the degree of injury to the corneal epithelium.

Comparison of serum cardiac troponin-Ⅰ and creatine kinase MB isoenzyme concentrations in asphyxiated neonates

Objective:To assess the correlation of signs of myocardial damage to serum cardiac troponin I(cTnI)and creatine kinase MB isoenzyme(CK-MB)concentrations.Methods:Blood samples were collected from 25 term asphyxiated neonates and 25 controls at 12 h of age by immunoassay.The asphyxiated neonates were followed up until discharge or death.Results:Asphyxiated neonates had significantly higher concentrations of cTnI and CK-MB than controls(P<0.001).Serum cTnI concentrations were significantly higher in asphyxiated neonates who developed hypotension,heart failure or those had low ejection fraction(P<0.01).Serum cTnI concentrations were significantly higher in asphyxiated who died than those who survived(P<0.01).There was no significant difference in serum CK-MB mass concentrations between asphyxiated neonates with and without these complications.Conclusion:Unlike CK-MB,serum cTnI concentrations are significantly higher in asphyxiated neonates who died or developed cardiac dysfunction.

Lactate Dehydrogenase Isoenzyme of Lens in Fetus

Purpose: The LDH isoenzyme patterns were evaluated in lens of fetus to study the metabolism in human lenses.Methods: LDH isoenzyme electrophoresis was performed in 15 lenses of human fetus aged from 4～6 months.Results: There are 5 bands of LDH in fetus lens. LDH4 lever is the highest, and LDH1 is the lowest, especially in the cortex of lens. LDH1 is higher in lens of 4 months of age than in that of 5 and 6 months of age.Conclusions: LDH isoenzyme in lens of fetus shows mainly anaerobic metabolism in development, but there is also aerobic metabolism, especially in younger fetus.