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Z Mishal

Publications and source records attributed to Z Mishal.

102 records · Page 6Linked to original sources
PubMed1981

The effect of hexamethylene-bis-acetamide on multiparameter analysis of Friend leukemia cells.

Friend leukemia cells were induced to differentiate by hexamethylene-bis-acetamide (HMBA). The relationship between cell growth cell volume distribution, fluorescence intensity and fluorescence anisotropy of the membrane probe 1,6 diphenyl 1,3,5 hexatriene (DPH) was analyzed in differentiated and undifferentiated cells with the aid of the fluorescence activated cell sorter (FACS II). The induction of a differentiated state was associated with a decrease in cell volume and an increase of fluorescence anisotrophy. The inhibition of this induction by dexamethasone prevented the decrease of cell volume without affecting the increase of fluorescence anisotropy. We conclude therefore that the high degree of anisotropy is not correlated to the differentiated state nor with the volume of the cells.

Acetamides↗
PubMed1981

Differentiation of thymocytes during chicken ontogeny: occurrence of a specific DNA ligase in relationship to cell size and surface antigens.

The activities of two forms (8.2 and 6.2 S) of DNA ligase have been studied in chicken thymocytes at different stages of development (16 and 18 days of incubation, hatching and one month after birth). Thymocytes have been sorted out on the basis of size and T antigenicity (immunofluorescence) with the activated cell sorter. Three kinds of cells can be distinguished: (a) large, antigenically negative cells of 16 days of incubation, expressing only the "heavy" (8.2 S) form of enzyme. The activity of this form decreases during development and is very low after birth; (b) large, antigenically positive cells without DNA ligase activity and (c) small, antigenically positive cells, expressing the "light" (6.2 S) enzyme. This form of enzyme appears only from 18 days of incubation onwards. These results allow us to consider the 6.2 S DNA ligase activity as an additional marker (with cell reduction and T antigenicity) for thymocyte maturation.

Aging↗
PubMed1981

Steady-state fluorescence polarization (FP) of 1,6-diphenyl-1,3,5-hexatriene (dph)-labelled platelets in the presence of bradykinin (bk).

Steady-state fluorescence polarization (FP) decreases, when 1,6-diphenyl-1,3,5-hextriene (dph) labelled platelets are exposed to bradykinin (bk). At pH 8, the dose-response curve is bell-shaped with an optimum bk effect at 10(-7) M. In contrast to the ricinoleic-acid ester of glycerin-polyethyleneglycol, cremophor EL (CEL), bk is no more effective when platelets are pretreated with 10(-5) M p-bromophenacylbromide (B phi B). These results suggest that platelets are target cells for the peptide bk, which induces an FP decrease indirectly by stimulating the release of non-saturated fatty acids in the platelet membrane.

Animals↗
PubMed1981

Effect of pH on fluorescence polarization of DMSO induced Friend leukemic cells.

The effect of DMSO on Friend leukemic cell membranes was investigated in an inducible cell line (FLC) and in cell variant resistant to DMSO induction (RFLC). Fluorescence polarization (P) studies carried out at 37 degrees C with diphenyl hexatriene (DPH) have shown that P values change according to the pH, being maximum in alkaline buffer. These changes were altered when cells were grown in the presence of DMSO. Changes in P values according to the pH were abolished by glutaraldehyde and by sodium azide. Fluorescence polarization analysis of FLC cell line carried out at 4 degrees C have shown higher P values which also changed according to the pH. In DMSO induced FLC or in RFLC grown with or without DMSO, these changes were not seen. It is therefore concluded that P value changes related to pH reflect changes in the lipid-protein interactions of cell surface membrane. It is also suggested that these interactions are altered by DMSO.

Animals↗
PubMed1978

Decreased microviscosity of membrane lipids in leukemic cells: two possible mechanisms.

Steady-state fluorescence polarization studies with the fluorescent lipid probe 1,6-diphenyl 1,3,5-hexatriene were done to determine the degree of microviscosity of cellular membrane lipids and serum lipoproteins in human normal donors and leukemic patients. The results show a marked decrease in microviscosity of cellular membrane lipids in both intact lymphocytes and isolated cellular plasma membranes obtained from leukemic patients in clinical relapse as compared to intact lymphocytes and isolated cellular plasma membranes obtained from normal donors and leukemic patients in complete clinical remission. Concomitant to these dynamic changes in cellular membrane lipids, the degree of microviscosity of lipids in the blood serum of leukemic patients in clinical relapse is markedly reduced as compared to serum obtained from normal donors and leukemic patients in complete clinical remission. Moreover, an in vitro incubation of leukemic lymphocytes with normal low density lipoproteins results in an increased microviscosity of cellular membrane lipids. In addition to the interrelation between cellular membrane lipids and serum lipoproteins, plasma membrane vesicles with a high degree of lipid microviscosity were isolated from the blood serum and pleural effusion of leukemic patients in clinical relapse. Such membrane vesicles could not be detected in normal serum. Therefore, we suggest that the two major mechanisms associated with the decreased microviscosity of membrane lipids in human leukemic cells are an abnormal exchange in lipids between the leukemic cell surface membrane and leukemic serum lipoproteins and an exfoliation of plasma membrane vesicles with a high degree of microviscosity from the cell surface of leukemic cells.

Cell Membrane↗
PubMedDate not supplied

Modulation of TNF-mediated cell lysis in vitro: further analysis of intracellular signaling.

We have investigated the post-receptor events governing the Tumor Necrosis Factor (TNF)-mediated cytotoxicity in vitro. As calcium has been reported to be an essential mediator in the cell killing processes, we asked whether an early increase in intracellular calcium could be involved during TNF-induced cell death. Using the ACAS methodology (adherent cell analysis and sorting), we could not detect any significant increase in intracellular calcium following TNF treatment (40 s) within the TNF-sensitive human breast carcinoma MCF7 cell line. In addition, A23187 (0.1-0.4 microM) did neither enhance TNF-mediated MCF7 cell lysis, further confirming that TNF-mediated cell lysis can occur in the absence of an early calcium increase. Given the potentiating effect of cAMP-inducing agents, such as forskolin, on TNF-mediated cytotoxicity, we have investigated the relationship between cAMP accumulation and the TNF signaling pathway during cell death. Our results indicate that the potentiating effect of forskolin (50 microM) on TNF-mediated MCF7 cell lysis did not involve a modulation in the TNF-induced activation of the nuclear factor NF-kB but was associated with an increase in the DNA fragmenting capacity of TNF as assessed by agarose gel electrophoresis of target cell DNA.

Adenocarcinoma↗
PubMedDate not supplied

Mineralocorticoid hormone signaling regulates the 'epithelial sodium channel' in fibroblasts from human cornea.

We investigated the regulation of sodium absorption by steroid hormones in embryologically diverse cells from the human eye. A cell extract from human corneal fibroblasts was positive for both the epithelial sodium channel (ENaC) and the mineralocorticoid receptor (MCR) as 82- to 85-kD and 102-kD bands, respectively, by the Western blot technique. In fluorescent, confocal and electron microscopy, the MCR was revealed as a nucleocytoplasmic protein, whereas the ENaC was almost exclusively membrane bound; both appeared aligned along actin filaments of corneal keratocytes, and both were widely colocalized in various cell types of human cornea in situ. Following reverse transcription and amplification of total RNA isolated from corneal fibroblasts, the ENaC and MCR genes in the PCR product were evident as predicted bands of 520 and 843 bp, respectively, whose sequence exhibited 100% identity with those from known human sources. The multiplication of corneal fibroblasts was influenced by both the MCR-specific antagonist RU 26752 and the natural hormone aldosterone, and these steroids also stimulated protein phosphorylation. In quantitative PCR, both the basal and aldosterone-induced levels of ENaC were diminished by the MCR-specific antagonist ZK 91587. Consequently, the ocular sodium channel appears to be regulated by steroid signalling in cells of diverse embryological origins, contrary to the existing notions where (a) this process would be limited exclusively to the epithelial cells and (b) ocular sodium transport would be regulated via the Na(+)-K(+)-ATPase in the basolateral membrane.

Aldosterone↗
PubMedDate not supplied

Mineralocorticoid hormone receptor and the epithelial sodium channel in a human leukemic cell line.

A Cell extract from the HEL (human erythroblastic leukemia) cell line was positive for both the epithelial sodium channel (ENaC) and the mineralocorticoid receptor (MCR) as glycosylated 82-84 kDa bands, and a single 102 kDa band, respectively, in Western blots using polyclonal antibodies raised against these proteins. The immunofluorescent labeling of the MCR in all cell lines showed a nucleocytoplasmic localization of the receptor whereas the ENaC was exclusively membrane-bound. These results were confirmed by confocal microscopy. The expression of the MCR in HEL cells was evident as a predicted band of 843 bp (234 amino acids) after total RNA from HEL cells had been reverse transcribed and then amplified by PCR; the ENaC was similarly evident as a predicted band of 520 bp. In both cases, near 100% identity was observed between the deduced amino acid sequences of the PCR products and those from known human sources. The multiplication of HEL cells was influenced by antagonists (RU 26752, ZK 91587) targeted for specificity to the MCR and this was reversed by the natural hormone aldosterone. These steroids also provoked chromatin condensation in the HEL population.

Amino Acid Sequence↗
PubMedDate not supplied

Changes in biophysical parameters and in phospholipid composition associated with resistance to doxorubicin.

Friend leukemia cells (FLC) resistant to different concentrations of doxorubicin were used to investigate the biochemical and biophysical changes associated with resistance. We have found that fluorescence polarization of 1,6-diphenyl-1,3,5-hexatriene analyzed on single cell level increased in resistant as compared to sensitive FLC. Furthermore, phospholipid analysis of sensitive and doxorubicin-resistant cells revealed changes in ratios of phosphatidyl-choline to phosphatidyl-ethanolamine and phosphatidyl-choline to sphingomyelin. These results correlate with decreased electrophoretic mobility in resistant cells. Our results indicate that changes in cell structure occur with the level of resistance to doxorubicin. These changes are probably the consequence rather than the cause of resistance.

Cells, Cultured↗