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[Identification of gravity-related esterases (grEST1 and grEST2) in carrot callus cells].

A horizontal clinostat which mimics the microgravity in space was used to study its effects on carrot cells. After using boric acid buffer (pH 8.8) as extraction medium, PAGE pattern of esterase isozymes of carrot callus cells displayed 8 bands of which the activities of only 2 were affected by microgravity. They were named grEST1 and grEST2, where grEST stands for gravity-related esterase. The rates of increase in activity of them in carrot cells when cultured on a rotating horizontal clinostat were lower than that cultured in normal gravitational environments, and the difference increased with the culture time. The activities of grEST1 and grEST2 in carrot callus cells subjected to horizontal rotation were found to return to their original levels after being placed under normal gravity (1 x g) for 7 days. We suggest that the effect of simulated microgravity conditions on grEST1 and grEST2 activities in carrot callus cells is through affecting their synthesis. In addition, the activities of grEST1 and grEST2 were not inhibited by eserine, acetylcholine iodide, diisopropyl fluorophosphate and p-chloromercuribenzoate, which indicates that they are acetylesterases.

Acetylcholine↗

[Characters of two gravity-related esterases in carrot callus cells].

On the basis of identification of two gravity-related esterases (grEST1 and grEST2) in carrot callus cells (Cai et al. 1998), we continued the study of the characteristics of these two esterases. They have the very special characteristic of SDS resistance. Their activities could be inhibited partially by deoxycholate. beta-Phenylpropionic acid, AgNO3 and CuSO4 had no inhibitory effect on their activities. The activities of grEST1 and grEST2 could be decreased by ascorbic acid and cysteine, and the influence by cysteine was particularly obvious. The molecular weights of grEST1 and grEST2 were shown to be near the ranges of 49-66 kD and 43-59 kD respectively by non-denaturing electrophoresis containing deoxycholate, Triton X-100 and SDS respectively, and the isoelectric points were approximately pH 5.4 and 4.9 respectively. Besides, grEST1 and grEST2 were found in the fraction precipitating at value between 30% and 40% saturation with (NH4)2SO4.

Copper Sulfate↗

[2 new esterase loci Es-14 and Es-15 in mice (genus Muc L.) : characterization by different substrates and inhibitors].

Allelic differences at an esterase locus designated Es-14 exist between Mus musculus (both inbred strains and wild Mice) and Mus spretus. This locus is expressed in a variety of tissues. Mus musculus has been assigned the allele Es-14r while Mus spretus carries the alternate alleles Es-14l. Each phenotype manifests itself as a single band of enzyme activity and F1 hybrids show two bands corresponding to the parental forms. Allozyme variation was also detected at another esterase locus, temporarily disignated as Es-15 until confirmed by current linkage analysis. A slow variant Es-15 was recorded in a Mus musculus population from Greece while inbred strains carry the Es-15m allele. Study of a Mus spicilegus population from Greece revealed the presence of a third allele, Es-15r. Substrate and inhibition characteristics are provided for both loci.

Alleles↗

Identification and characterization of novel organophosphate detoxifying esterase alleles in the Guangzhou area of China.

In the mosquito Culex pipiens, various alleles at the Ester locus provide insecticide resistance. These resistance alleles display a heterogeneous geographical distribution, particularly in China, where they are highly diverse. A new resistance allele, Ester9, coding for the overproduced esterases A9 and B9, is characterized and compared to the known resistant allele Ester8 isolated from the same southern China sample (from Guangzhou). Both alleles provide low but significant resistance to chlorpyrifos (relative synergism ratio [RSR] > 3) and temephos (RSR = 1.4), which is consistent with the low level of gene amplification they display (15 copies for Ester9 and 4 copies for Ester8). The full genomic sequence of the allele coding A8 and A9 is presented, which allowed us to set up a polymerase chain reaction assay to specifically identify these alleles. The peculiar situation in southern China, where numerous resistance alleles coexist, is discussed in comparison with the Mediterranean situation, the only one with a similar diversity of overproduced esterases.

Alleles↗

[Adaptation of microtitration plate methods for quantification of the activity of esterases and glutathione-s-transferase in Aedes aegypti].

The detection techniques for the activity of non-specific esterases and glutathione-s-transferase in Culex quinquefasciatus were modified to detect such enzymes in Aedes aegypti(L). The optimal concentration values of substrate (saturating concentration) and the optimum reading time for reaction were determined for each enzyme by using 4 Aedes aegypti strains: one from Cuba 2 from Venezuela and one susceptible reference strain. The frequency of non-specific esterases turned out to be 0.76 in MIRANDA 0.42 in ARAGUA and 1 in SANTIAGO DE CUBA in which the highest frequency value of this mechanisms was reached. The frequency of glutathione-s-transferase mechanism was 0.45 in ARAGUA 0.043 in MIRANDA and 1 in SANTIAGO DE CUBA. For the first time in Cuba a set of biochemical techniques was available for detecting the resistance mechanisms of Aedes aegypti which made it possible to create sound foundations for the easy rapid detection of the resistance of this species the main dengue vector in the Americas.

Aedes↗

[Allozyme variation of nonspecific esterases in Russian sturgeon (Acipenser güldenstädti Brandt].

Electrophoresis in 7% polyacrylamide gel was used to analyze nonspecific esterases from different organs and tissues of Russian sturgeon. The basic enzymatic activity was observed the four zymogram zones. In sixteen cases the patterns of isozyme distribution in these zones allowed to consider them as allele systems (14 polymorphic and 2 monomorphic) corresponding to the four structural genes. The observed genotype frequencies of the nine of these systems (EST-1* of brain and intestine; EST-2* of serum and intestine; EST-3* of spleen and kidney; and EST-4* of spleen, brain, and heart) were largely concordant with Hardy-Weinberg proportions for codominant disomic inheritance. Genetic control of other polymorphic systems is unknown, but, high enzymatic activity of these esterases, sufficient for qualitative electrophretic detection, permits utilization of these polymorphisms for phenotypic monitoring of Russian sturgeon populations.

Animals↗

[Polymorphism of water-soluble and triton-X 100-extractable esterases and proteins from Leptospira].

Electrophoresis in gel from polyacrylamide was used to study the water-soluble intracellular esterases, triton-X 100-extracted and proteins of three saprophytic and three pathogenic strains of leptospirae belonging to different serological types. The results of investigation of a relative mobility of 27 fractions with an esterasic activity and the molecular weight of eight of them showed that their polymorphism was mainly consecuent of structural differences. In examination of proteins extracted from the cell residues with triton-X 100 there was also revealed an intertype polymorphism: by acrylamide electrophoresis with sodium dodecylsulphate they were divided into 3--10 fractions with a molecular weight of from 20000 to 200000. The results of these studies are discussed as a manifestation of biochemical individuality of leptospirae of individual serological types.

Bacterial Proteins↗

Two-dimensional electrophoresis patterns of total proteins, esterases and acid phosphatases from pollen, seeds and leaves of three cultivars of Helianthus annuus L.

Total proteins, esterases and acid phosphatases from pollen, seeds and leaves of three sunflower cultivars were separated by 2-D electrophoresis. The characteristic peptides for each cultivar were identified. The seeds and pollen of the cultivar Wielkopolski contained 45 and 37 characteristic peptides, respectively, while the seeds and pollen of Coril contained 73 and 35 characteristic peptides. The cultivar Frankasol had the lowest total number of stained peptides in seeds and pollen, and the number of the characteristic peptides was only 61 and 25, respectively. Analyses of esterases and acid phosphatases from young leaves and pollen led to identification of isoenzymes characteristic of the three cultivars. Only for Frankasol no specific acid phosphatase was observed, both in leaves and in pollen.

Acid Phosphatase↗

Esterase as molecular marker for salt tolerance in regenerated plants of rice, Oryza sativa L.

Esterase variation was studied in plants regenerated from callus cultures of four rice (Oryza sativa) varieties, viz. pokkali, which is a moderately salt tolerant variety and three salt sensitive varieties MI 48, annapoorna and jyothi. Variation was studied at tillering stage of plants regenerated from callus culture and germinated from seeds. Somaclonal variants for salt tolerance could be detected using variation in esterase banding pattern and activity.

Biomarkers↗

Prognostic value of flow cytometrically determined DNA-ploidy, intracellular pH and esterase activity of non-small cell lung carcinomas.

30 surgical specimens of patients with non-small cell lung carcinomas (NSCLC) were investigated. Significant increases of intracellular pH values in epithelial and inflammatory cells, in the percentage of dead epithelial and inflammatory cells and in the cell volume of vital inflammatory cells in cancerous lung tissue were encountered. Furthermore, decreases of the esterase activity of vital epithelial cells and of the percentage of free cell nuclei were observed. The DNA aneuploidy in 36.6% of the tumours was frequently associated with non-squamous cell carcinomas and stage II, III, IV tumours. Patients with DNA aneuploid tumours had a significantly shorter survival rate than those with DNA euploid tumours. Within the different tumour stages a similar tendency was observed which was, however, only significant in stage III tumour patients. Stage III tumours constitute therefore a heterogeneous entity with a worse prognosis for DNA aneuploid tumour patients. The intracellular pH values and esterase activity as well as the cell volume, the percentage of free cell nuclei and dead inflammatory or epithelial cells contained no significant prognostic information.

Adult↗

Acne vulgaris. Studies in pathogenesis: suppression of nonspecific esterases.

Oral tetracycline eliminated the histochemical staining for nonspecific esterase in human sebaceous glands after two weeks of administration. This evidence offers further support for the hypothesis that the clinical benefits of tetracycline result from suppression of follicular esterase-lipases, probably those from C. acnes.

Acid Phosphatase↗

Blood esterase determinations as markers of exposure.

The bases of using blood enzyme activity measurements [e.g. AChE, non-specific cholinesterase (BChE), carboxylesterase] as markers of organophosphate ester (OP) exposure are inhibition of activity by the binding of OPs to serine active sites in the enzymes, and the accessibility of the enzymes in RBCs and serum. The methods used to determine esterases in the blood of humans, experimental animals, and wildlife are outlined with emphasis on the acetylcholinesterase (AChE) of the red blood cell. Adaptations of an acetylthiocholine ester assay of Ellman et al. (1961) are common, but other colorimetric procedures, radiometric assays, and pH methods are also in use. Optimized, standardized methods are needed to assess exposures and provide a solid basis for risk assessment analyses. Useful adjuncts to ChE measurements are oxime reactivation tests and assay of neuropathy target esterase, an enzyme associated with organophosphate-induced delayed neuropathy. Determination of urinary metabolites compliments, but does not substitute for, the information obtained from blood ChE studies. Future assays are likely to involve antibodies to OP-protein complexes. Improvements in techniques permit the detection of small decreases in ChE activities. Whether or not such small decreases in ChE activities can, by themselves, constitute an adverse effect for input into risk assessment analyses is a controversial matter.

Biomarkers↗

[Determination of esterase activity of human and animal serine proteinases using fluorogenic esters of amino acids as substrates].

Two fluorogenic derivatives of amino acids are proposed as substrates for the purpose of enzymatic assay: N-benzyloxycarbonyl-phenylalanine-4-methyl umbelliferyl ester (substrate-1) and tert-butyloxycarbonyl-alanine-4-methyl-umbelliferyl ester (substrate-II). Chymotrypsin-like (hydrolysis of substrate-1), elastase-like (hydrolysis of substrate-II) esterase activity of bovine pancreatic chymotrypsin, activities of cathepsin G and elastase from human, porcine and rat neutrophils and esterase activity of human, porcine and rat serum were assayed. Differences in the level of chymotrypsin-like and elastase-like activities of human, porcine and rat serum were established. Activities of purified elastase and cathepsin G from human and animal neutrophils were shown to have no significant distinctions.

Animals↗

[Isoenzyme spectrum of lactate dehydrogenase, malate dehydrogenase, esterase and acid phosphatase of rat brain cells at different times after external 1 Gy gamma irradiation].

The influence of external single gamma-irradiation with a dose of 1 Gy on the isoenzyme composition of lactate dehydrogenase, malate dehydrogenase, esterase and acid phosphatase in the cytoplasm of rat brain cells has been investigated. Irradiation was shown to cause differently directed changes in the ratio of the isoenzymes under study at different times after exposure. The isoenzyme spectrum of lactate dehydrogenase and malate dehydrogenase was shown to be normalized on day 30 after irradiation, whereas the isoform composition of esterase and acid phosphatase was not stabilized at that time.

Acid Phosphatase↗

[Characterization of Alcaligenes species using analysis of esterase electrophoretic polymorphism and analysis of antibiotic resistance profiles].

The species of an Alcaligenes bacterial strain may be difficult to determine on the basis of conventional phenotype features. Esterase pattern analysis using acrylamide-agar gel electrophoresis and determination of the antimicrobial resistance profile (agar diffusion method) were performed for A. faecalis (34 strains). A. denitrificans subsp xylosoxydans (16 strains) and A. piechaudi (5 strains). The Cistat program (D2 Software) was used for statistical representation of results. The homogeneous, species-specific esterase patterns ensured correct assignment of each strain to one of the three species. Antimicrobial susceptibility was greatest for A. faecalis which was susceptible to both cephalosporins of all generations and aminoglycosides. A. xylosoxydans was the species with the greatest resistance to antimicrobials. A. piechaudii exhibited intermediate susceptibility.

Alcaligenes↗

Performance of Spodoptera litura Fabricius on different host plants: influence of nitrogen and total phenolics of plants and mid-gut esterase activity of the insect.

Five host plants [castor, Ricinus communis (Carolus Linnaeus); cotton, Gossypium hirsutm (Carolus Linnaeus); tomato, Lycopersicum esculentum (Philip Miller); mint, Mentha arvensis (Carolus Linnaeus) and cabbage, Brassica oleracea (Carolus Linnaeus)] belonging to different families were used to study the performance of the Asian armyworm, Spodoptera litura larvae. Highest consumption of food and dry weight gain was observed in larvae fed on castor. Mint did not support optimum larval growth because of low digestibility and low efficiency of conversion of digested food to body matter. Dry weight gain ranged from 26.64 mg on mint to 86.80 mg in castor. These differences tend to be related to nitrogen and total phenolics content of the leaf tissues; however, the most clear-cut correlation is an inverse one between the host plant preference and the ratio of total phenolics to nitrogen in the leaf tissues. Mid-gut esterase activity in larvae showed an increasing trend with the increase in total phenolics: nitrogen ratio in the test plants and the order of mid-gut esterase activity in larvae was mint > cabbage > cotton > tomato > castor.

Animals↗

Hydrolysis of organophosphorus compounds by an esterase isozyme from insecticide resistant pest Helicoverpa armigera.

Esterase activity of resistant and susceptible H. armigera were compared in gels with different substrate such as naphthyl acetate, naphthyl phosphate, paraoxon and monocrotophos. Whole body extract of resistant H. armigera hydrolyzed paraoxon, monocrotophos and naphthyl phosphate in gels. Resistant H. armigera showed high esterase, phosphatase and paraoxon hydrolase activity compared to susceptible ones.

Animals↗