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E Bar

Publications and source records attributed to E Bar.

15 recordsLinked to original sources

Expression of chromosomally inserted bacillus thuringiensis israelensis toxin genes in bacillus sphaericus

Bacillus thuringiensis israelensis delta-endotoxin genes were inserted into transposon Tn917 in plasmid pTV51Ts and cloned into the chromosome of Bacillus sphaericus 2362. Many of the transformants reacted with antibody to the 135-, 128-, 65-, and 28-kDa B.t.israelensis toxin proteins and were approximately 10 times more toxic to A. aegypti larvae than the untransformed host. Some of the transformants differed physiologically and morphologically from the wild-type B. sphaericus. The toxicity of the transformed phenotype was maintained through many transfers in the absence of selective pressure. Copyright 1998 Academic Press.

Journal Article↗

Assessment of DNA damage induced in vitro by etoposide and two fungicides (carbendazim and chlorothalonil) in human lymphocytes with the comet assay.

The effects of two fungicides (carbendazim and chlorothalonil) on the induction of DNA damage in human peripheral blood lymphocytes (human PBL) have been investigated using the single cell gel electrophoresis assay (SCGE assay or comet assay) immediately after a 1-h treatment and after a 24-h post-treatment incubation. The assessment of etoposide (an effective antitumour agent) effects on human PBL in terms of cell viability and dose-DNA damage relationships was made and etoposide selected as a positive control. The results indicate that etoposide induces significant (p < 0.01) dose-dependent DNA damages for concentrations at which the loss of cell viability is low. After a 24-h recuperation period, all observed DNA damages has disappeared. With SCGE assay performed after a 1-h treatment, similar positive results were observed with chlorothalonil alone or in association with carbendazim, without any loss of cell viability. However, a dramatic loss of cell viability was measured after 24 h and was associated with a large proportion of highly damaged cells. In contrast, carbendazim was not cytotoxic on human PBL and did not induced DNA damage using the SCGE assay either immediately after treatment or after a 24-h post-treatment incubation. These results point to the necessity of an adequate evaluation of immediate and long-term cytotoxicity of compounds that are to be assessed by the SCGE assay.

Adult↗

Serum galactosyltransferase activities in newborn children and pregnant women.

We have compared serum galactosyltransferase activity in pregnant women and in newborn children. Subjects in this study were 75 women at different stages of pregnancy and 60 newborns, including premature infants. Activity ratios in pregnant women are based on the period of gestation, expressed in weeks of amenorrhea. Average values were 265 nmol/h/ml for up to 12 weeks of amenorrhea, 369 from 13 to 28 weeks and 483 over 28 weeks versus 232 nmol/h/ml for non-pregnant women. In infants, galactosyltransferase activity decreased with increasing age from conception, but the activity level was always much higher in newborns than in women at the ninth month of pregnancy. We discuss the origin of the enzyme in these various samples.

Female↗

Biological markers and ovarian carcinomas: galactosyltransferase, CA 125, isoenzymes of amylase and alkaline phosphatase.

We present a comparative study of several biological markers (galactosyltransferase, CA 125, isoenzymes of amylase and alkaline phosphatase) with a view to ovarian carcinoma follow-up. Serum samples were obtained from a population of 75 patients under clinical observation. After a minimum 18-months period, we assessed the prognostic value of the markers. No marker permits the detection of discrete, evolving carcinomas. CA 125 is the marker that gives the best results, particularly in terms of sensitivity. Galactosyltransferase has a lower sensitivity except in the case of endometrioid carcinomas. Simultaneous analysis with CA 125 and galactosyltransferase results in no decisive improvement, other than greater precision in unfavourable prognoses. Isoenzymes of amylase and alkaline phosphatase are of no interest in the follow-up of such carcinomas.

Alkaline Phosphatase↗

Cloning and expression of Bacillus thuringiensis israelensis delta-endotoxin DNA in B. sphaericus.

Bacillus thuringiensis israelensis delta-endotoxin genes were cloned into Bacillus sphaericus 2362, producing stable transformants reacting with antibody to the 28- and 65-kDa B. thuringiensis israelensis crystal proteins and approximately 10 times more toxic to Aedes mosquito larvae than the original host strain. The LC50 after 48 hr of exposure of Aedes larvae to the most active transformed clone was 0.19 microgram/ml, compared with an LC50 of 1.9 microgram/ml for B. sphaericus 2362 and less than 0.1 microgram/ml for B. thuringiensis israelensis. The cloning vector, plasmid pPL603E, was also effective in transforming B. subtilis 1E20 with B. thuringiensis israelensis DNA, producing highly toxic clones with less stable gene expression than the clones of B. sphaericus.

Aedes↗

[Localization and modulation of galactosyltransferase during in vitro proliferation of a human ovarian adenocarcinoma cell line].

A cell line, IGROV1, originating from a human ovarian cancer, releases a galactosyltransferase activity in its culture medium during proliferation. The proliferating IGROV1 cells appear as two populations: some cells grow in floating clusters whereas the greater part of them adhere to the culture substrate. The study of galactose transfer by intact cells onto an exogenous glycoprotein acceptor (ovomucoid) shows the presence of surface-associated galactosyltransferase onto the two cellular sub-populations. Opposite to intracellular activity, surface-associated and released galactosyltransferase activities depend on cellular adhesion and proliferation.

Adenocarcinoma↗

A simple serum galactosyltransferase assay method suitable for routine use.

UDP-Galactose: N-acetylglucosaminyl glycoprotein beta 1-4 galactosyltransferase (GT) catalyzes the transfer of galactose to N-acetylglucosamine from UDP-[3H]Gal. The uncharged reaction product (tritiated N-acetyllactosamine) is separated from the unreacted UDP-[3H]Gal by ion-exchange chromatography. The major advantage of this method is its rapidity compared to other isotopic techniques.

Chromatography, Ion Exchange↗

Alteration in isoenzyme patterns of serum galactosyltransferase activity in ovarian cancer patients: preliminary results.

Isoelectric focusing on agarose gel was used to separate the isoenzymes of serum galactosyltransferase (uridine diphosphogalactose: N-acetylglucosaminyl galactosyltransferase, EC 2.4.1.22) from 8 healthy women, and 11 ovarian cancer patients of whom 4 were in clinical remission. In all cases, we found 7 major peaks with isoelectric points ranging from 4.0-5.4. The most acidic peaks were preferentially elevated in the tumor-bearing patients, particularly the peak with pI 4.44.

Aged↗

Serum galactosyltransferase activity with three acceptors in ovarian cancer patients.

Several laboratories have demonstrated the usefulness of serum galactosyltransferase as a biological marker for ovarian neoplasms. However, contradictory results have been published recently, which might be partially explained by differences in methodology. We thus decided to measure serum galactosyltransferase activity in patients with ovarian cancer and benign gynecological diseases using three different assay systems. A very good correlation was obtained between the results of these assays. Furthermore, we confirm that serum GT is frequently elevated in cancer patients, and is of potential value for their follow-up.

Acetylglucosamine↗

[Comparative study of various technics for the determination of human serum galactosyltransferase].

We have compared three assays for serum galactosyltransferase activity, which use ovomucoïd, asialo agalactofetuin and free N-acetylglucosamine respectively as exogenous acceptors. A very good correlation between the three assays is obtained, for the whole range of GT activity. When the methods are compared to one another, the slopes of the regression lines are similar whether the sera are collected from healthy controls, pregnant women, or women suffering from benign gynecologic diseases or ovarian neoplasms. The methods which uses free N-acetylglucosamine as acceptor is rapid, cheaper and easier to perform.

Female↗

[Separation of isoenzymatic forms of human serum galactosyltransferase by chromatofocusing and isoelectric focusing. Application to cancerology].

Chromatofocusing is used to separate the multiple isoenzyme forms of human serum galactosyltransferase. At least 11 major peaks of activity are observed in normal sera, which are eluted between pH 4.3 and 6.9; a fraction of activity is eluted above pH 7.0. The normal patterns are compared with those obtained with sera from cancer patients and with an ascitic fluid. Chromatofocusing appears as resolutive as agarose isoelectric focusing.

Ascites↗

[Determination of human serum galactosyltransferase using a kinetic spectrophotometric technic].

A kinetic spectrophotometric method in which galactose transfer is coupled to the production of NADH, has been adapted to the assay of galactosyltransferase activity in human serum. Under the described conditions, the rate of NADH production is linear with regard to enzyme concentration, and directly depends upon the various biochemical factors which control galactosyltransferase activity.

Acetylglucosamine↗

Isoenzymatic patterns of human galactosyltransferase from normal and cancer patients.

Chromatofocusing revealed the high heterogeneity of human serum galactosyltransferase with regard to isoelectric point. At least 11 major peaks of activity were observed for normal sera, at the following pH elutions: 4.3, 4.4, 4.6, 4.77, 5.0, 5.15, 5.45, 5.8, 6.35, 6.65, and 6.9. In addition, a fraction of the activity was eluted above pH 7. Some of those peaks were also present in sera from cancer patients, but almost all the peaks in the 7.4-5.4 pH range had disappeared. Alpha-lactalbumin affinity chromatography strongly modified the chromatofocusing pattern of galactosyltransferase from an ascitic fluid of a cancer patient.

Chromatography, Affinity↗