PubMed Health⌕ Search

Biomedical subjects

Z Végh

Publications and source records attributed to Z Végh.

At least 19 recordsLinked to original sources

Cleaning validation procedure eased by using overpressured layer chromatography.

In the manufacturing plants of many pharmaceutical companies the reaction apparatus is suitable to produce different active pharmaceutical ingredients. After completing the production of a compound the equipment should be cleaned in order to avoid the cross contamination in the next lot of the other products. In the authors' laboratory several chromatographic methods were introduced to measure the amount of the residual substances remaining on the surface of the apparatus after the cleaning procedure. A sensitive and fairly rapid overpressured layer chromatographic (OPLC) procedure--suitable to separate and control five steroid hormone compounds (allylestrenol, estradiol, ethynodiol diacetate, levonorgestel, norethisterone) produced in the same equipment at different times--was developed and validated.

Chromatography, Liquid↗

[The effect of prolonged acenocoumarol therapy on bone density].

The effect of chronic cumarin treatment on bone mineral content was investigated. Bone mineral density was determined by double photon densitometry (Lunar DPXL). The density data (mean +/- SE) of 45 cardiac patients (age: 57.0 = +/- 6.3 y, body mass index: 26.7 +/- 3.8 kp/m2, cardiac stadium score, according to New York Heart Association: 2-3), had been treated by acenocumarol at least for 2 years (duration of treatment: 75.0 +/- 52 months), were compared to the values of 45 age, body mass index, cardiac status matched patients not treated by anticoagulant. The density values of L2-L4 lumbar regions were lower in the treated group (1.041 +/- 0.17 vs. controlls: 1.13 +/- 0.15 g/cm2, p < 0.05), while no differences in ultradistal ulnar and radial regions were detected. No correlation between bone mineral density and the length, or the dose of the cumarin treatment were observed. This observation suggests the importance of the regular bone densitometry control of cumarin treated patient.

Acenocoumarol↗

Cytotoxic susceptibility and defective MHC class I expression do not correlate with mutation of p53 in human carcinomas.

Twenty-nine samples of ex vivo ovarian and lung carcinomas were investigated for the relationship between the presence of mutated protein 53 (mp53) and cytotoxic susceptibility. Unaltered expression of MHC class I alleles was required for the cytotoxic susceptibility of tumour cells to the autologous ex vivo blood lymphocytes, i.e. all 4 sensitive tumours belonged to the group of 11 tumours without defect in MHC class I expression. In contrast, the susceptibility did not correlate with the presence of mp53, i.e. cases with mp53 were randomly distributed between the sensitive and resistant tumours (2/4 and 10/17 respectively). There was no correlation either between the p53 mutation and down-regulation of MHC class I alleles. The results suggest that in these tumours the mutated p53 is not the source of immunogenic peptides and that the lack of recognition of the tumours with mp53 is not caused by a defect in the expression of MHC class I molecules.

Adenocarcinoma↗

Generation of HLA-B55 restricted T lymphocyte mediated cytotoxicity against autologous LCL.

The lymphocytes of one HLA-A11 positive individual (A1, A11, B49, B55) were stimulated in vitro with the autologous EBV transformed lymphoblastoid cell line (LCL). The culture contained HLA-A1, A11 and B55 restricted, LCL selective cytotoxic T cells (CTLs). In the T cell culture stimulated four times, the lysis of A11 and B55 carrying targets suggested that the subsets of the two latter CTL types had similar size. After further stimulations the B55 restricted CTLs were enriched in the culture. Earlier results suggested that in HLA-A11 positive individuals the A11-restricted LCL-selective CTL subset dominates. The sensitivity of a target panel including Burkitt lymphoma (BL) lines suggested that the peptide presented by the B55 molecule differs in A and B type EBV strain carrying cells.

Cell Line, Transformed↗

Selectively down-regulated expression of major histocompatibility complex class I alleles in human solid tumors.

We studied the expression of major histocompatibility complex (MHC) class I molecule in 52 ex-vivo tumor samples comprising 29 ovarian, 15 lung, 1 breast, and 4 colon carcinomas; 1 midgut carcinoid; and 2 malignant mesenchymal tumors obtained from surgical specimens or from malignant effusions. The allelic products were visualized in untreated and interferon gamma + tumor necrosis factor alpha treated aliquots of tumor cells and in the patient's blood lymphocytes by the one-dimensional isoelectric focusing method. Generally, the tumor cells contained lower amounts of MHC class I molecules compared to the lymphocytes. In vitro exposure to interferon gamma and tumor necrosis factor alpha elevated the level of MHC class I expression in 24 of 52 tumors and corrected the assembly defect seen in 2 cases. In 20 tumors one or several human leukocyte antigen alleles were undetectable even after cytokine treatment. Correlation was seen with the grade of differentiation; the proportion of tumors with selective losses in poorly, moderately, or well differentiated tumors were 16 of 30, 3 of 13, and 1 of 9, respectively. Selective losses occurred in ovarian carcinoma cells collected from malignant effusions (12 of 22, 54%) but not in 7 primary tumors. In primary lung carcinomas the frequency was 36% (5 of 14 cases). Thirty-nine patients were serologically typed; thus the MHC alleles on the tumor cells could be identified. In this panel of tumors 30 human leukocyte antigen alleles were represented. Among these the expression of 15 was found to be down-regulated in some but not all tumors of the same histology.

Alleles↗

Increased expression of MHC class I molecules on human cells after short time IFN-gamma treatment.

Human cell lines and blood lymphocytes were treated for short time periods with IFN-gamma. This treatment increased the amount of the assembled MHC class I molecules on the plasma membrane after 30 min. This early increase of the membrane expression subsided in the next few hours. A second wave of elevation occurred after 8-24 hr. Analysis of cytoplasmic and membrane molecules in pulse chase experiments showed that the cytokine enhanced both the assembly of available heavy and light chains and the transport of the complex to the plasma membrane. The membrane level of the HLA-A2 molecules showed similar kinetics. Addition of an A2 specific binding peptide stabilized the IFN-gamma induced molecules on the cell surface. It seems that IFN-gamma alone or together with a binding peptide can influence MHC class I expression solely through post-transcriptional events utilizing an available pool of free heavy and light chains already after a short time, before the enhancement of the synthesis starts.

Amino Acid Sequence↗

Red-cell bound anti-A is more efficient than anti-B in competition for fluid phase complement.

Lysis of group A and B erythrocytes by human complement was studied by an anti-A (BRIC.131) and an anti-B (BRIC.30) IgM monoclonal antibody in a 51Cr-release assay. The relative concentration of membrane-bound immunoglobulins was detected by flow cytometric analysis, and the amount of C1q and C3 bound to the sensitized red cells was measured by using purified, 125I-labelled molecules. The direct haemolysis was identical with both reagents in the presence of excess and suboptimal complement over a wide range of antibody concentration (between 50 and 7000 ng/ml). The indirect effect of membrane-bound antibody, i.e. its influence on complement binding by sensitized bystander cells, was examined in a cold target competition assay in which sensitized, non-labelled cells are present when complement is incubated with sensitized labelled cells. We have found that the competitive capacity of sensitized erythrocytes correlated with the amount of membrane-bound immunoglobulins. In accordance with our earlier findings, an equal level of target and competitor cell lysis was obtained only if the fluid phase anti-B antibody concentration was 2 to 4 times higher than that of the anti-A antibodies. We demonstrate in this paper that the different competitive activity of IgM anti-A and anti-B monoclonal antibodies might be accounted for by differences in their C1q and C3 binding capacities.

ABO Blood-Group System↗

Interactions in the complement-mediated lysis of blood group AB erythrocytes sensitized simultaneously with anti-A and anti-B monoclonal antibodies.

The lysis of group AB erythrocytes by human complement was studied by different anti-A and anti-B IgM monoclonal antibodies (mabs) in a 51Cr-release assay. The concentration of membrane-bound immunoglobulin was detected by ELISA, and the amount of C1q and C3 bound to sensitized red cells was measured by using purified, 125I-labelled molecules. We have demonstrated that there is an exponential relationship between the concentration of the sensitizing IgM mabs and C1q binding to the sensitized AB cell. The efficiency of binding was related to the number of antibodies bound; thus, anti-A sensitized cells bound 3-6 times more C1q than anti-B sensitized cells did. AB cells, on the other hand, bound similar amounts of C3 whether anti-A or anti-B was present. The lytic efficiencies of the various IgM mabs during short incubation times were different, suggesting that the complement activation rates vary widely with different antibodies on the AB cell membrane. The binding of C1q to an antibody-sensitized target activates a cascade, whose components may migrate away from the sensitizing antibody; interactions between the activation processes generated by the anti-A and anti-B antibodies may thus occur. Choosing appropriate pairs of anti-A and anti-B mabs for the simultaneous sensitization of AB cells has indeed resulted in stimulation in some and inhibition in other combinations of mabs. It is suggested that stimulation is observed when the activated intermediates are produced in excess, whereas inhibition occurs when a shortage of activated intermediates prevents mutual utilization.

ABO Blood-Group System↗

Assembly of MHC class I molecules in ex vivo carcinoma cells induced by IFN-gamma or by a binding peptide.

It has been reported that the assembly of MHC class I molecules in mutagenized cell lines could be induced by specific binding peptides. We have now demonstrated that the defect in assembly between heavy and light chains of class I molecules naturally occurred in tumor cells of one spontaneous ovarian carcinoma detected by one-dimensional isoelectric focusing of immunoprecipitates with anti-monomorphic class I MAb (W6/32) and by immunostaining with free heavy chain and beta 2m-specific MAbs. In vitro treatment of the tumor cells with IFN-gamma induced the assembly and surface expression of majority class I molecules (A2.1, B7, B15, Cw6, Cw7 out of A2.1, A2*, B7, B15, Cw6, Cw7). Moreover, assembly of A2 and Cw6 was induced by exposure of the tumor cells to a HLA A2-binding peptide K62 derived from influenza A matrix protein. Autologous blood T lymphocytes were activated in mixed lymphocyte-tumor cell culture (MLTC) by the IFN-gamma-treated but not by the unmanipulated tumor cells. Although activated lymphocytes damaged both IFN-gamma-treated and untreated tumor cells, the alpha class I MAb (W6/32) efficiently inhibited the lysis of IFN-gamma-treated targets, but not the untreated targets. These results indicate that the defect of MHC class I assembly may result in the escape of tumor cells from immune response.

Amino Acid Sequence↗

[Analysis of steroids. Part 45: Analytical investigation of pipecuronium bromide (Arduan)].

The following methods are described for the analytical investigation of pipecuronium bromide. 1. HPLC method. Of the several systems tried for the separation and quantification of impurities and degradation products the best results were obtained using silica as the stationary phase and 43:43:14 mixture of methanol, acetonitrile and concentrated aqueous ammonia containing 0.1 mole/l each of ammonium chloride and ammonium carbonate as the eluent. The validation of this method is presented. The above described aggressive eluent can be successfully replaced by an ion-pairing system using silica as the stationary phase and 96:4 mixture of acetonitrile and water containing 0.1 mole/l sodium perchlorate as the eluent. 2. Thin-layer chromatography. TLC systems are described for the separation and densitometric quantification of the impurities and degradation products of pipecuronium bromide. 3. Spectrophotometry. Two methods are described. The ester groups of the molecule can be determined by the iron(III)-hydroxamate method while for the ion-pair extraction of the quaternary ammonium steroid picric acid or bromthymol blue are used as the reagents. 4. Titrimetry. In addition to the titration with acetous perchloric acid for the assay of the bulk material a microtitration method is described for the determination of pipecuronium bromide in individual lyophylized ampoules (potentiometric titration with 0.1 M silver nitrate).

Androstane-3,17-diol↗

A re-evaluation of the lipid-bound sialic acid determination.

The origin of serum sialic acid measured in the lipid-bound sialic acid determination reported by Katopodis et al. (1980) was investigated in detail. By varying the experimental conditions of sample preparation the protein, lipid and sialic acid contents of the methanol-water extract obtained from human sera were analyzed and compared in healthy controls and cancer patients. Using polyacrylamide gel electrophoretic and gel chromatographic methods it has been shown that most of the lipid-bound sialic acid was attributed to the acid alpha 1-glycoprotein (orosomucoid) fraction of human sera. Based on these observations a re-evaluation of the molecular background of the LBSA determination seems to be necessary.

Blood Proteins↗

[Color coded duplex scanning examination of the carotid artery].

A recent developed ultrasound technology that combines the B-mode real time imaging, colour flow imaging and pulsed Doppler velocity measurements with spectral analysis was used for bilateral carotid artery examinations of 81 patients with suspicion of carotid stenosis. The results in 9 cases was compared with angiographic findings. This paper describes the basic principles of this method, the fundamental of the estimation of the lumen reduction. Authors present the advantages and limitations of this method and conclude that the colour coded duplex scanning sonography is an accurate noninvasive technique that suitable for detection of the extracranial carotid artery diseases. They emphasize that in all of 5 cases with amaurosis fugax significant homolateral stenosis was found in the communis or internal carotid artery.

Blindness↗

Expression of MHC-class-I antigens in human carcinomas and sarcomas analyzed by isoelectric focusing.

We analyzed the expression of class-I antigens in ex vivo human tumor cells by isoelectric focusing (IEF) the anti-class-I mAb W6/32 immunoprecipitates prepared from cell lysates. Out of 42 experiments, 27 were technically successful. The patient's blood lymphocytes were used as controls. In vitro exposure of the tumor cells to IFN gamma and TNF alpha elevated class-I antigen expression. In 11 cases, defects in MHC-class-I-antigen expression were observed. In 2 cases the antigens were detected only in the cytokine-treated tumor samples, probably due to a defect in the association between beta 2m and class-I heavy chains. Selective changes in the expression of alleles were seen in 10 cases and might involve HLA A, B and C antigens. Alterations in class-I expression as compared with the lymphocytes were observed in 9 of 13 cases in which the tumor cells were collected from metastases, and only in 2 of 14 primary tumors.

Adenocarcinoma↗

[Correlation of slime production and pathogenicity of coagulase-negative staphylococci].

Coagulase-negative staphylococci have been increasingly recognized as pathogens in patients with indwelling medical devices. Slime production may be an important virulence factor. Slime production and antibiotic susceptibility were studied of 117 coagulase-negative staphylococci isolated from clinical specimens. The frequency of slime production among 26 coagulase-negative staphylococci was about three times higher than among 91 isolates regarded as contaminants. The former strains were isolated from 10 patients, 8 of which had device related infections. Multiply resistant cultures occurred in 22.2%, but all strains were susceptible to vancomycin.

Coagulase↗

Monoclonal free light chains in urine and their significance in clinical diagnostics: are they really tumor markers?

Bence Jones proteins (monoclonal free light chains of immunoglobulins) are the earliest known biological markers of malignant cell dyscrasia; Bence Jones proteinuria is also present in many types of B cell-related neoplasms. Sometimes, it may also occur in Hodgkin's disease. In some cases, benign monoclonal gammapathy was found to be associated nontumorous diseases as well. The type of monoclonal light chain, the degree of polymerization, and the isoelectric point of the molecule may affect the course of the disease. Urine samples from 637 patients with true or suspected lymphoproliferative diseases were investigated over a 2-yr period by different immunochemical methods. Bence Jones proteinuria was identified in 71 cases by isoelectric focusing combined with immunofixation, while the pathological protein was detected only in 63 cases by conventional methods. Bence Jones proteins can be detected by this new method at a level below the sensitivity of conventional procedures. Bence Jones proteins in the urine may signal a malignant tumor or malignant transformation of an earlier disease. The early detection of monoclonal immunoglobulin light chains in the urine may be important in clinical diagnosis, therapy, and follow-up.

Bence Jones Protein↗

Identification and cDNA cloning of a new nodule-specific gene, Nms-25 (nodulin-25) of Medicago sativa.

A new nodule-specific gene, Nms-25 (nodulin-25), was identified in cDNA clones isolated from a nodule-specific cDNA library of Medicago sativa. The first transcript of this gene appeared 9 days after inoculation of the roots with Rhizobium meliloti. The time of expression and the quantity of the transcripts of the Nms-25 gene was similar to that of leghemoglobin genes suggesting a similar regulation. A protein of 246 amino acids could be deduced from a full-length cDNA clone. The first 24 amino acids at the N-terminal end of this protein formed a signal sequence which might direct membrane transport into the peribacteroid space. Using different predictive methods the signal sequence cleaved protein was tentatively predicted to be a water-soluble enzyme, but not hydrolase.

Amino Acid Sequence↗

The nucleotide sequence of a nodule-specific gene, Nms-25 of Medicago sativa: its primary evolution via exon-shuffling and retrotransposon-mediated DNA rearrangements.

We present the primary structure of a nodule-specific gene, Nms-25 from Medicago sativa L. cultivar Nagyszénási. Analysis of the nucleotide sequence of Nms-25 revealed that this gene shows all the characteristics of an interrupted plant gene consisting of 13 exons and 12 introns. The promoter region of Nms-25 contains the common promoter elements of plant genes as well as motifs which are supposed to be involved in nodule-specific expression. There are two exon-like sequences in the gene named PE1 and PE2 which are not present in the cDNA clones of Medicago sativa cultivar Cardinal. Intron 9 carries a retrotransposon-like element, Tms1, which might be responsible for downstream deletion events in which a heptanucleotide, ATTAGCT, might have been involved. Most of the exons, except 1, 12 and 13, are similar to each other both in length (54 bp) and sequence (up to 94% sequence similarity). All exons are interrupted by introns in the same phase (type I). It is suggested that exon-shuffling based on illegitimate recombination in which the ATTAGCT motif might have played an active role, and retrotransposon-mediated DNA rearrangements were the primary events in the molecular evolution of the Nms-25 gene.

Amino Acid Sequence↗