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

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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.

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.

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.

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.

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.

Clinical and preclinical treatment of urologic diseases with phosphodiesterase isoenzymes 5 inhibitors： an update

Phosphodiesterase isoenzymes 5 inhibitors （PDE5-1s） are the first-line therapy for erectile dysfunction （ED）. The constant discoveries of nitric oxide （NO）/cyclic guanosine monophosphate （cGMP） cell-signaling pathway for smooth muscle （SM） control in other urogenital tracts （UGTs） make PDE5-1s promising pharmacologic agents against other benign urological diseases. This article reviews the literature and contains some previously unpublished data about characterizations and activities of PDE5 and its inhibitors in treating urological disorders. Scientific discoveries have improved our understanding of cell-signaling pathway in NO/cGMP-mediated SM relaxation in UGTs. Moreover, the clinical applications of PDE5-1s have been widely recognized. On-demand PDE5-1s are efficacious for most cases of ED, while daily-dosing and combination with testosterone are recommended for refractory cases. Soluble guanylate cyclase （sGC） stimulators also have promising role in the management of severe ED conditions. PDE5-1s are also the first rehabilitation strategy for postoperation or postradiotherapy ED for prostate cancer patients. PDE5-1s, especially combined with （z-adrenoceptor antagonists, are very effective for benign prostatic hyperplasia （BPH） except on maximum urinary flow rate （Qmax） with tadalafil recently proved for BPH with/without ED. Furthermore, PDE5-1s are currently under various phases of clinical or preclinical researches with promising potential for other urinary and genital illnesses, such as priapism, premature ejaculation, urinary tract calculi, overactive bladder, Peyronie＇s disease, and female sexual dysfunction. Inhibition of PDE5 is expected to be an effective strategy in treating benign urological diseases. However, further clinical studies and basic researches investigating mechanisms of PDE5-1s in disorders of UGTs are required.

3种虾类超氧化物歧化酶同工酶比较(英文)

[Objective] The aim of the study is to compare the activities of superoxide dismutase(SOD) isoenzymes in three species of shrimps Penaeus japonicus, Procambarus clarkia and Litopenaeus vannamei. [Method] The experimental materials were used to measure SOD activities after pretreatment, meanwhile the differences in SOD isoenzymes from different materials were assayed by polyacrylamide gel electrophoresis (PAGE). [Result] There are specific and histological differences in SOD activities of shrimps. With a similar electrophoresis pattern and migration rate, Penaeus japonicus and Litopenaeus vannamei showed remarkable differences with that of Procambarus clarkia. [Conclusion] The result showed the differences of cognation and origin of three shrimps.

Expression of 5α-Reductase Type 2 Gene in Human Testis, Epididymis and Vas Deferens

Objectives To study the expression pattern of 5α-reductase type 2 gene in human male reproductive organsMethods The expression level of 5α-reductase type 2 gene inhuman testis, epididymis and vas deferens tissues was determined by in situ hybridization using Digoxin labeled 5α-reductase type 2 cRNA probe.Results The brown granules of hybridizing signals distributed in the cytoplasm of Sertoli and Leydig cells of the testis, the principle cells of epididymis and the epithelial cells of vas deferens, but there was no positive signal in the nuclei of above-mentioned cells. No positive signal was observed in germ cells, basement of the testis, interstium of epididymis and basement, as well as smooth muscle of vas deferens. Conclusion This study confirmed that the 5α-reductase type 2 gene expressed in Sertoli , Leydig cells of the testis, and the principle cells of epididymis. The expression pattern of the gene in these cells in human was similar to that of rat and monkey. The presence of 5α-reductase type 2 gene in epithelial cells of the vas deferens suggested it might possess an important physiological role in human reproduction.

5α-reductase type 2 gene expression in human testis, epididymis and vas deferens

To study the expression pattern of 5α-reductase type 2 gene in human male reproductive organs. Methods: The expression level of 5ct-reductase type 2 gene in human testis, epididymis and vas deferens tissues was determined by in situ hybridization using a digoxin-labeled 5α-reductase type 2 cRNA probe. Results: The brown granules of hybridizing signals distributed in the cytoplasm of the Sertoli and Leydig cells of the testis, the principle cells of epididymis and the epithelial cells of vas deferens, but there was no positive signal in the nuclei of these cells. No positive signal was observed in the germ cells, basement of the testis, interstium of the epididymis and basement and the smooth muscle cells of vas deferens. Conclusion: This study confirmed that the 5a-reductase type 2 gene expressed in the Sertoli and Leydig cells of the testis and the principle cells of the epididymis. The expression pattem of the gene in these cells in the human was similar to that in the rat and monkey. The presence of 5α-reductase type 2 gene in the epithelial cells of the vas deferens suggests that it may play a physiological role in human reproduction. [Reprod Contracep (in English) 2002; 13: 67]

Biochemical variability between two Egyptian <i>Stenodactylus </i>species (Reptilia: Gekkonidae) inhabiting North Sinai

Polyacrylamide gel electrophoreses for malate dehydrogenase (Mdh) and beta-esterase (β-Est) isoenzymes were conducted for biochemical differentiation between two Stenodactylus gekkonid species inhabiting North Sinai of Egypt. Total lipids and proteins of liver and muscle tissues in both species were also analyzed. A total of three Mdh isoforms were recorded in the analysis, in which the activity of Mdh-2 and Mdh-3 seemed to be higher in S. petrii than in S. sthenodactylus. This high activity could be supported by the significant increase in the total lipids and proteins in liver and muscle tissues of the species. It may thus be reasonable to suppose that S. petrii is more active, energetic and adaptable in the desert habitat than S. sthenodactylus. β-Est showed six fractions in S. petrii and only one fraction in S. sthenodactylus. It is therefore noticeable that β-Est is more highly expressed in S. petrii than in S. sthenodactylus.

Genetic and Metabolic Variability between Two Subspecies of <i>Chamaeleo chamaeleon</i>(Reptilia: Chamaeleonidae) in Egypt

The degree of variability between two subspecies of Chamaeleo chamaeleon;C. chamaeleon chamaeleon inhabiting El-Dabaa (MarsaMatrouh) and C. chamaeleon musae inhabiting El-Arish (North Sinai) of Egypt was investigated in this study using polyacrylamide gel electrophoreses for Lactate dehydrogenase (Ldh) and Alfa-esterase (α-Est) isoenzymes. Total lipids and proteins of liver and muscle tissues in both species were analyzed. Three Ldh isoforms were recorded for both subspecies and the activity. Rate of flow (RF) of Ldh-1 seemed to be higher in C. chamaeleon chamaeleon than in C. chamaeleon musae. This high activity could be supported by the significant increase in the total lipids and proteins in liver and muscle tissues of this species. It may thus be reasonable to suppose that C. chamaeleon chamaeleon is more active, energetic and adaptable in its habitat than C. chamaeleon musae. The α-Est showed four fractions in both subspecies. The null variations in the activity of α-Ests in the studied tissue may indicate, to some extent, the safety of the diet applied to both subspecies of chameleons.

Cancer and pH—A Prospective

How can cancer develop in so many different organs in so many different ways, but the outcome is similar enough to all be under the same title—cancer? There are so many causes of cancer—viruses, genetic defects, sunburn, gastroesophageal reflux, smoking, alcohol, radiation, chemicals, etc. The above variable well known etiologies of cancer could all induce a need for repair which involves alkalinizing the cells involved. Thus the commonality for cancers could be a pH change. If true, this could give the field of cancer prevention and therapy new avenues of pursuit.

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.

The nervous system and pH

The mix of cellular pumps, channels, transporters, and isoenzymes is genetically inherited. However, the environment has the ability to impact the cellular pumps, channels, transporters, and isoenzymes to some degree. Cellular pumps, channels, transporters and isoenzymes help control cellular pH and vice versa. Therefore possibly the pH in the nervous system is more variable than previously believed. If so, that could explain some of the changes we see in the nervous system.

Preliminary Comparative Physiological and Biochemical Study of Five Different Goat Breeds Inhabiting Saudi Arabia

Three arbitrary chosen enzymes were examined by native-polyacrylamide gel electrophoresis to investigate physiological and genetic variations among five different goat breeds inhabiting Saudi Arabia. The goat breeds were Pakistani, Tihami, Syrian, Masri and Aardi while the investigated enzymes were alkaline phosphatase (ALP), malate dehydrogenase (Mdh) and malic enzyme (ME). Six polymorphic loci with six monomeric alleles have been recorded in all studied breeds. The second locus of ME was characteristic of Syrian breed where it showed dimeric alleles. The similarity matrix that has been calculated according to the number of sharing bands indicated that these breeds have been divided into two groups: Pakistani and Tihami as one group while the other three breeds clustered in another group. The activity of the metabolic enzymes (Mdh and ME) showed concordance with the constructed relationship where the percentage amounts of these enzymes showed significant variations between the two groups more than that occurred within each group. Mdh was expressed in the second group more than in the first while ME showed, nearly, equal expression in the different breeds. Both genetic and physiological results agreed in clustering the studied breeds into Pakistani and Tihami in one group and the other three breeds in another group. This division was based on a few gene loci and a few sampling size and it needs to be supported by collecting more genetic data and more sampling size in a further molecular study.

Stimulating Effects of Seed Treatment by Magnetized Plasma on Tomato Growth and Yield

Tomato seeds （Lycopersicon esculentum L. Mill. cv. zhongshu No. 6） were treated by magnetized plasma before being sown to investigate its effect on the growth and yield of tomatoes. Biochemical analysis showed that dehydrogenase activity increased with the increase of the current but decreased when the current was higher than 1.5 A. The activities of peroxidase （POD） isoenzyme changed in the same pattern. There was no difference in germination percentage between treatments and control, which were carried out in laboratory conditions. However, significant （c~ = 0.01） difference was observed in germination percentage in the pot experiment. In the pot experiment, the sprouting rate for the treatment with a 1.5 A current was 32.75%, whereas the untreated was only 4.75% on the eleventh day. Germination time is more than one day earlier than the control. The 1.5 A treatment increased the tomato yield by 20.7%.

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.