Acid Phosphatase Patterns Research Articles

Salinity acclimation alters acid and alkaline phosphatase expression and histological changes in the hepatopancreas of the oriental river prawnMacrobrachium nipponense(De Haan, 1849) (Decapoda: Caridea: Palaemonidae)

AbstractThe oriental river prawn, Macrobrachium nipponense (De Haan, 1849), an economically important species, can live in freshwater and brackish water environments, but molecular mechanism of adaptation to varying salinity levels is still poorly understood. We aimed to investigate the effects of four different salinities (0, 6, 12, and 18‰) on metabolic responses, histological structure, and expression patterns of acid phosphatase (ACP) and alkaline phosphatase (AKP) in the hepatopancreas. The full-length cDNAs of ACP and AKP of M. nipponense (designated MnACP and MnAKP, respectively) were obtained by rapid-amplification of cDNA ends (RACE)-PCR. Immunofluorescence experiments confirmed that MnAKP is located in the cell membrane of the hepatopancreas. Both genes were expressed in all seven studied tissues, with highest expression in the hepatopancreas by quantitative real-time PCR. The mRNA expression levels and enzyme activities of MnACP and MnAKP in the hepatopancreas significantly increased in 18‰ salinity group compared to other groups. Measurements of glycogen, triglycerides, and total cholesterol levels suggested that prawns can adjust the energy metabolism in a way to provide additional energy under unsuitable salinity. Besides, prawns at high salinity produced higher B-cells and lower number of R-cells in hepatopancreatic tubules, also suggesting enhance the metabolic function of the hepatopancreas under high salinity. Our results provide a better understanding for the mechanism regulating salinity acclimation in the oriental river prawn.

Effect of Lufenuron and Oriza sativa Bran Extract on Fraction Protein and Acid Phosphatase Pattern in Haemolymph of Schistocerca gregaria.

The extensive use of insecticides has significant negative aspects such as the development of insect resistance, great threat to environmental health, and increased costs. IGRs and compounds from natural origin considered environmentally safe and less hazardous to non-target biota. So, the current study aimed to evaluate the insecticidal activity of the match (lufenuron) and rice bran (Oriza sative) extract and their effect on fraction protein and acid phosphatase (ACP) pattern of haemolymph of treated Schistocerca gregaria. The present findings cleared that, LC50s estimate for one-day old of 5th nymphs of S. gregaria were 35.66 and 541.71 ppm for lufenuron and O. sativa bran extract, respectively. As to fraction protein pattern, a maximum number of 65 bands were detected, at molecular weight ranged from 6.73 to158.31and 18 unique bands were scored. The epigenetic distance accounted for 1 for lufenuron and 0.76 for Oriza sativa bran extract, 20 days post-treatment. In addition, seven acid phosphatase bands were recognized in treated and untreated 12 samples, with Rf values ranged between 0.02 and 0.86. Three unique bands were scored. In conclusion, the treated nymph with the lethal concentration LC50 of the tested compounds (lufenuron and O. sativa bran extract) showed a significant change in protein fraction and acid phosphatase patterns of haemolymph, which was clearly reflected in high epigenetic differences between control and treated S. gregaria samples.

Virulence of Entomopathogenic Nematodes Steinernema glaseri and Heterorhabditis bacteriophora Poinar (HP88 strain) Against the Black Cutworm, Agrotis ipsilon.

Virulence of entomopathogenic nematodes, Steinernema glaseri and Heterorhabditis bacteriophora Poinar (HP88 strain) were studied against 3rd, 4th, 5th and 6th instar larvae of the black cutworm, Agrotis ipsilon. The observed mortality caused by the both tested nematodes at different time intervals was recorded. The establishment of nematodes in a host depends greatly on its ability to manage host defences so, the host haemocyte interaction with entomopathogenic nematodes were observed by encapsulation, nodule formation and phagocytosis processes. Phagocytosis is known to stimulate production of lysosomal enzymes. Acid phosphatase acts as a lysosome marker so; the activity and the pattern of acid phosphatase during the course of infection were recorded. Also, total protein and protein pattern were screened. Results indicated that, the two nematodes had variable significantly effects. S. glaseri recorded a highly significantly affect than H. bacteriophora. In addition, there are relationship between mortality rate and time exposure of S. glaseri and H. bacteriophora against 3rd, 4th, 5th and 6th larval instars of A. ipsilon i.e. the time exposure increased the susceptibility also increased. At the lower concentration, fourth and/or fifth instars were the most susceptible instars to S. glaseri ,while the 3rd and 4th instar larvae were most vulnerable instars to the infection with H. bacteriophora. Results was recorded that, six haemocyte types were identified in A. ipsilon 6th instar larvae: prohaemocytes, plasmatocytes, granulocytes, spherulocytes, oenocytoids, and adipohaemocytes. S. glaseri in A. ipsilon escape encapsulation and overcome host immunity faster than H. bacteriophora which recognized by host haemocytes. Prohemocytes, plasmatocytes or granulocytes are phagocytic cells in A. ipsilon 6th instar larvae. Phagocytosis was expressed in A. ipsilon haemocytes infected with H. bacteriophora at a higher rate compared with S. glaseriinfection. Data demonstrated both S. glaseri and H. bacteriophora induced a significant decrease in total protein contents after 24hrs from infection. While, there was significant increase in acid phosphatase activity after 6 hrs of infected groups of S. glaseri and H. bacteriophora. This increase may be due to phagocytosis processes which recorded in the present study at this time because phagocytosis is known to stimulate production of lysosomal enzymes. There is an acid phosphatise band detected only in H. bacteriophora and S. glaseri samples with Rf 0.86 after 6 and 12hrs from infection. This band may refer to nematodes exudates which contain high amount of this enzyme that the nematodes can easily penetrate and digest host tissues. There is a characterestic band of S. glaseri protein pattern with Mw 47KDa. It was identified as surface coat protein of S. glaseri which play important roles in defeating the host immune system by its antiphagocytic activity.

The Effects of Phosphate Starvation on the Activities of Acid and Alkaline Phosphatase, Fructose-1,6-bisphosphatase, Sucrose-phosphate Synthase and Nitrate Reductase in Melon (Cucumis melo L.) Seedlings

Plants response to phosphate starvation include the changes of activity of some enzymes, such as phosphatases, fructose-1,6-bisphosphatase, sucrose-phosphate synthase and nitrate reductase. In this study, to determine the effects of phosphate starvation on the change of activities of acid and alkaline phosphatase, fructose-1,6-bisphosphatase, sucrose-phosphate synthase, and nitrate reductase were studied in melon seedlings (Cucumis melo L.). The content of the protein and chlorophyll tended to relatively reduced in melon seedlings subjected to phosphate starvation. Acid phosphatase activity in first and second leaves of melon seedlings was relatively higher than that of third and fourth leaves of seedlings in 14 days after phosphate starvation treatment, respectively. Active native-PAGE band patterns of acid phosphatase in melon leaves showed similar to activities of acid phosphatase, whereas alkaline phosphatase activity was different from the change in the activity of acid phosphatase. Inorganic phosphate content in melon seedlings leaves was constant. The changes of Fructose-1,6-bisphosphatase and sucrose phosphate synthase activities showed similar patterns in melon seedlings leaves, and between these enzymes activities and phosphate nutrition negatively related. Fructose-1,6- bisphosphatase and sucrose phosphate synthase activities showed significant difference in second and fourth leaves, but nitrate reductase showed significant difference in first and second leaves in 14days after phosphate starvation treatment. We concluded that phosphate nutrition could affect the distribution of phosphate, carbon and nitrogen in melon seedlings.

Acrylamide gel electrophoresis of proteins, acid phosphatases and RN-ases from three potato varieties

Studies on variety differences in the protein and acid phosphatase patterns as well as ribunuclease activity distribution were carried out by disc electrophoresis on saline extracts of three varieties of the potato <i>Solanum tuberosum</i> (L.). The protein bands varied in number, position and relative abundance. One main zone of the acid phosphatase activity was detected consisting of 2-3 electrophoretically different bands. Variety differences were concerned with the number and relative abundance of these bands. RNase activity was detected in 4 main zones, in some of them additional subbands were visible. Differences between the three examined varieties were reflected in the occurence of the particular activity zones or their subbands.

Total protein and isozyme characterization in the flax zygotic embryo during development

Flax zygotic embryogenesis was studied for isozyme patterns of acid phosphatase and esterase and for content of total proteins. For acid phosphatase, six multiple molecular forms or isozymes, were identified during flax zygotic embryo development. For esterase, six isozymes were expressed during zygotic flax embryogenesis. Some of the isozymes were expressed during entire embryogenesis, and some were only transiently expressed or appeared near maturation. The amount of total proteins was very low at early embryogenesis and proteins in a range from 26.6 kDa to 97.4 kDa were expressed. During further flax embryo development the protein spectra became more diverse and the quantity of total proteins increased. Proteins ranging from 3.4 kDa to 97.4 kDa were present, and proteins of 20 kDa to 36.5 kDa were expressed in the highest amount. The results are discussed with reference to selected information in the literature.

Análises isoenzimáticas em somaclones de morangueiro (fragaria x ananassa Duch.) cv. Vila Nova

Análises isoenzimáticas foram utilizadas para investigar variações bioquímicas em somaclones de morangueiro cultivar Vila Nova, o btidos a partir de calos. Utilizaram-se como amostras folhas jovens de plantas durante a multiplicaç ão in vitro e após a aclimatização em casa de vegetação. Os sistemas enzimáticos utilizados para a análise dos clones foram fosfatase ácida e esterase (pH 6,5 e 8,3). Durante o cultivo in vitro, dois padrões de fosfatase ácida foram detectados entre os regenerantes; já para a esterase, todos os genótipos exibiram o mesmo padrão de bandas. Em casa de vegetação, foram encontrados três padrões de bandas distintos de fosfatase ácida. Não foram verificadas alterações quanto ao número e posição das bandas de esterase em pH 6,5, enquanto que para a esterase em pH 8,3 observou-se polimorfismo entre os clones. Ambos os sistemas enzimáticos mostraram-se influenciados pelas condições de crescimento (in vitro e in vivo) e pelos estádios de desenvolvimento das plantas.

Tetrad Analysis of Acid Phosphatase, Soluble Protein Patterns, and Mating Type inLeptosphaeria maculans

We investigated the utility of mating type, soluble protein profiles obtained after isoelectric focusing, and acid phosphatase (ACP) patterns as genetic markers for the blackleg fungus, Leptosphaeria maculans. Fertile matings were obtained in vitro by pairing single-ascospore isolates. A method for dissecting asci and recovering the eight component ascospores was developed that allowed tetrad analysis. Protein electrophoresis revealed limited polymorphism among the 73 single-ascospore field isolates. Only one isolate (H5) displayed a different soluble protein profile (SPP), termed SPP − , characterized by one missing band and one additional band. We found three ACP patterns in our collection. Protein profiles and ACP patterns were mitotically and meiotically stable

Enzyme Markers for Somatic Embryogenesis in Vitis

Summary A protocol has been detailed for somatic embryogenesis from leaf- and petiole-derived callus of Vitis rupestris. Isozymic patterns of acid phosphatase, alcohol dehydrogenase, esterase, gluconate-6-phosphate dehydrogenase and phosphoglucomutase were analyzed during precise steps of somatic embryogenesis from callus to plantlet. Among the five enzymes investigated, alcohol dehydrogenase, acid phosphatase and esterase gave different results, either in their specific activity, as in the case of alcohol dehydrogenase, or in the number and isoelectric point of the isoforms expressed, as in that of acid phosphatase and esterase. Non-morphogenic calli were clearly different from embryogenic calli, and from embryos. Moreover, the calli dedifferentiated from somatic embryos did not retain the level and types of isozymes that characterize the embryo state. Our analysis of the electrophoretic patterns of specific enzymes proves to be an effective approach to the characterization of the main steps of Vitis rupestris somatic embryogenesis.

Localization and isozyme patterns of phosphatase in Fibricola seoulensis

The present study was carried out to investigate the localization and isozyme patterns of acid phosphatase and alkaline phosphatase in metacercariae and in adults of F. seoulensis by enzyme-histochemistry method and electrophoresis. Acid phosphatase showed a strong activity at pH 5 in the intestinal caecum of adults, but showed no reactions in the non-substrate control and in the inhibitor-treated control. Alkaline phosphatase showed a strong activity at pH 8 in the intestinal caecum and the tribocytic organ of adults, and in the intestinal caecum and in the genital anlagen of metacercariae. In non-denature PAGE, ten bands of protein fraction from the extracts of metacercariae and twenty-two bands from adults were detected. In denature PAGE, two protein bands having molecular weights of 192 kDa and 123 kDa were detected in the metacercariae, but absent from adult stage. In adults, protein fractions of 27.5 kDa, 24.5 kDa, 21.4 kDa, 18 kDa, 16 kDa and 15 kDa were detected. In non-denature PAGE, isozymes of acid phosphatase showed the most strong activity at pH 5, whereas no activity was shown at pH 2 and pH 7. One isozyme band (62 kDa) of AcPase was recognized in metacercariae and 4 isozyme bands (95 kDa, 85 kDa, 73 kDa and 62 kDa) in adults.

Comparison of Isozyme Patterns between Spirometra erinacei and Spirometra mansonoides by Isoelectric Focusing

No differences were observed in the isozyme patterns of 4 enzymes examined between fresh samples stored at -80 C and samples stored at room temperature for 10 days after lyophilization, which supports the validity of comparing lyophilized samples to fresh frozen tissue. Mature proglottids as well as plerocercoids of Spirometra erinacei from Japan and Australia were indistinguishable by comparison of isozyme patterns after isoelectric focusing. The isozyme patterns of acid phosphatase, glucosephosphate isomerase (GPI), and mannosephosphate isomerase from plerocercoids of Spirometra mansonoides were distinctly different from those of plerocercoids of S. erinacei. The adenylate kinase isozyme patterns of the mature proglottids of S. mansonoides were also distinctly different from those of the mature proglottids and the plerocercoids of S. erinacei. The GPI isozyme pattern of the mature proglottids of S. mansonoides was also distinguishable from the GPI patterns of those of S. erinacei. These electrophoretic data suggest that the S. erinacei from Japan and Australia are closely related, if not identical, but that S. mansonoides is genetically distinct from S. erinacei.

The effects of a strong constant magnetic field on the activity and localization pattern of acid phosphatase in Blepharisma

The effects of a strong constant magnetic field on the activity and localization pattern of acid phosphatase in Blepharisma

FATTY ACID-SENSITIVE ACID PHOSPHATASE ACTIVITY OF TUBERCLE BACILLI

In a whole cell assay system with p-nitrophenyl phosphate as substrate, strains of Mycobacterium tuberculosis and M. bovis were identical in the pH-activity pattern of acid phosphatase. It was a one-peak curve with a pH optimum at 6.2 and sharp symmetrical slopes. The enzymatic activity did not reflect the virulence. When the cells were subjected to mechanical fractionation, the major part of the enzymatic activity was found in a particulate fraction and a minor portion in supernatant and cell walls, suggesting the location of the enzyme in the membrane. Exposure of the cells to free long-chain fatty acids, especially unsaturated ones, reduced the enzymatic activity in a dose-response manner with concomitant decrease in the viability. However, no causal relationship between these two effects was suggested from the collateral experiments.

Fetotoxic alterations in the normal ontogenies of rat microsomal and lysosomal enzymes

The activity patterns during development for acid phosphatase (Ac-P), alkaline phosphatase (A1-P), beta-glucuronidase (beta G), and UDP-glucuronyltransferase (UDPGT) have been determined in various tissues of the rat for corn oil and distilled water controls as well as in animals prenatally exposed to four fetotoxic chemicals. Postnatal assays were performed on both sexes separately. In control animals, tissue-specific differences between male and female activity levels were found for UDPGT. In the liver of mature offspring, enzyme activity was greater in males than in females. Although no sex difference was observed in the intestine, the kidneys of females exhibited higher values than those of males. An original computer-assisted methodology is presented, designed (a) to permit a mathematical description for the complex curves exhibited by these ontogeny profiles, and (b) to assess the statistical significance of chemical-induced alterations in these complex developmental patterns, specifically, to target sensitive periods and subtle changes near the fetotoxic threshold. Oral administration (days 6-18 of gestation) of 3,3',4,4'-tetrachlorobiphenyl (4CB) to pregnant females resulted in an induction of liver UDPGT activity in offspring postnatally, and some alterations in the perinatal pattern of beta G in the same tissue. This treatment also produced differences in the intestinal patterns of Ac-P and male UDPGT. No significant changes were observed in offspring exposed to diethylstilbestrol (DES). Treatment with zeranol (ZN) caused reductions in activity over the entire postnatal period for beta G in liver, brain, intestine, and kidney, for A1-P in brain, and for Ac-P in the intestine. Cadmium-treated dams gave birth to offspring that exhibited slightly altered ontogenies only in intestine for UDPGT and AcP. The alterations in these developmental profiles indicate periods of increased sensitivity, and may be useful in directing more specific studies into the fetotoxic mechanisms of these compounds.

Production of somatic hybrid between black mustard(Brassica nigra Koch.;BB) and Kakuran(B. napus L.;AACC).

Mesophyll protoplasts of black mustard and Hakuran were fused by the PEG-high pH ?? igh Ca method. Out of six plants regenerated, one showed hybrid morphology. The hybridity was ascertained by the isozyme pattern of acid phosphatase and chromosome number. This hybrid was estimated to be a trigenomic hexaploid having A. B and C genomes, and showed similar male sterility to Hakuran.

Acid phosphatases from liver of Cyprinus carpio

1. 1. Two acid phosphatase forms were isolated from liver extract of Cyprinus carpio using Sephadex G-200 chromatography. 2. 2. AcPase I has a molecular weight of about 118,000, is inhibited by NaF and tartarate, shows an optimal pH in the range 4.3–5.6 and hydrolases various phosphoric esters. 3. 3. AcPase II has a molecular weight of about 16,600, is insensitive to NaF and tartrate but is strongly affected by sulphydryl reagents, shows an optimal pH in the range 4.8–6.2 and the commonly used phosphatase esters are poor substrates for the enzyme. 4. 4. The effect of temperature indicates that the low molecular weight enzyme is more thermolabile than the high molecular weight form. 5. 5. Our results demonstrated that fish liver shows the same acid phosphatase pattern previously reported for birds and mammals.

Isozymes in nutrient medium of suspension-cultured apple cells (Malus sylvestris Mill.)

The time course of the activities of esterase, α-galactosidase, and β-glucosidase in cell sap and nutrient medium in in vitro cultured apple cells (Malus sylvestris Mill.) was studied. The corresponding isozyme patterns and the intracellular and extracellular isozyme patterns of acid phosphatase and polyphenol oxidase were compared using isozyme visualization methods adapted to ultra-thin-layer isoelectric focusing. Neither quantitative (total activity) nor qualitative (isozyme pattern) data were congruent for cell saps and nutrient media. Malate dehydrogenase, malic enzyme, and glutamate dehydrogenase occurred in cell sap only. The extracellular activities probably originate to a great part from a programmed release by intact cells. Nutrient media of plant cell cultures constitute a rich source of active plant isozymes.

Assays for total activity and isoenzyme pattern of acid phosphatase in urine from patients with renal diseases

Assays for total activity and isoenzyme pattern of acid phosphatase in urine from patients with renal diseases

Isozyme electrophoregrams of sixteen enzymes in five tissues of cassava (Manihot esculenta Crantz) varieties

Methodology, based on starch and polyacrylamide gel electrophoresis, was developed for determining isozyme electrophoregrams (patterns) of 16 enzymes of cassava (Manihot esculentaGrantz) varieties as potential genotype markers. Extracts of five different tissues (root, stem, leaf, petiole and bud) were examined. In general, the nodal portions of the shoots gave isozyme patterns with the largest number of bands. Petiole extracts gave similar results but bud extracts gave poor patterns. The limited number of varieties that were examined could be distinguished by sequential classification on the basis of the isozyme patterns of acid phosphatase, esterase, glutamate oxaloacetase transaminase and phosphoglucoisomerase.

Comparison of Acid Phosphatases in the Rat Prostatic Complex and Seminal Vesicles

Acid phosphatase activity of different parts (ventral, lateral and posterior lobes and coagulating gland) of the rat prostatic complex and seminal vesicles were analyzed in homogenate and after fractionation with gel filtration and chromatofocusing. Significant differences between the various tissue homogenates were recorded in the hydrolysis of p-nitrophenyl phosphate, α-naphthyl phosphate and naphthol ASBI phosphate and the percentage of the tartrate-resistant activity varied from about 30 (in seminal vesicles) to 56 (in coagulating gland). Gel filtration resulted in the appearance of 2 (Sephadex G-200) to 3 (Sepharose 6B) activity peaks (GF-1 to GF-3). Studies with 6 substrates (p-nitrophenyl phosphate, α-naphthyl phosphate, β-naphthyl phosphate, naphthol ASBI phosphate, phenolphthalein phosphate and thymolphthalein phosphate) showed some relative differences in the hydrolysis rates as well as indications for possible overlapping of multiple enzymes in these peaks. Chromatofocusing of the ventral prostate homogenate resulted in the appearance of 8 activities (CF-1 to CF-8) with different pi-values (8.2, 8.1, 7.9, 7.1, 6.4, 6.0, 5.5 and 5.0), substrate preferences, pH-optima (5.5, 4.2, 4.2, 6.0, 5.5, 5.0, 4.2 and 5.0), molecular weights and modifier characteristics. The other tissues contained a lower number of these activities: 4 (CF-1 to CF-3 and CF-6) in lateral and posterior lobes and coagulating gland and 5 (CF-1 to CF-4 and CF-6) in seminal vesicles. The correspondence of the enzyme peaks after gel filtration and chromatofocusing was analyzed by refractionation. This indicated that some of the enzymes (GF-1, CF-5, CF-8) may appear in polymeric/aggregate forms. The differences in the acid phosphatase pattern suggest characteristic functional features in the various parts of the rat prostatic complex.