scholarly journals Combined Cytogenotoxic Effects of Bee Venom and Bleomycin on Rat Lymphocytes: AnIn VitroStudy

2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
Yasmina M. Abd-Elhakim ◽  
Samah R. Khalil ◽  
Ashraf Awad ◽  
Laila Y. AL-Ayadhi
Keyword(s):  
Gradient Centrifugation ◽  
Fluorescent Microscopy ◽  
Molecular Techniques ◽  
Bee Venom ◽  
Genotoxic Effects ◽  
Quantitative Expression ◽  
Predominant Type ◽  
Rat Lymphocytes ◽  
Ldh Release ◽  
Isolated Rat

This study was carried out to determine the cytotoxic and genotoxic effects of bee venom (BV) and/or the chemotherapeutic agent bleomycin (BLM) on healthy isolated rat lymphocytes utilizing morphometric and molecular techniques. Using the Ficoll-Histopaque density gradient centrifugation technique, lymphocytes were isolated, divided into groups, and subjected to BV and/or BLM at incubation medium concentrations of 10 or 20 μg/mL respectively for 24 and 72 hrs. An MTT assay and fluorescent microscopy examinations were used to assess the cytotoxic effects. To determine the predominant type of BV and/or BLM-induced cell death, LDH release assay was employed beside quantitative expression analyses of the apoptosis-related genes (Caspase-3 and Bcl-2). The genotoxic effects of the tested compounds were evaluated via DNA fragmentation assay. The results of these assays demonstrated that BV potentiates BLM-induced cytotoxicity through increased LDH release and diminished cell viability. Nevertheless, BV significantly inhibited the BLM-induced DNA damage. The results verify that BV significantly attenuates the genotoxic effects of BLM on noncancerous isolated rat lymphocytes but does not diminish BLM cytotoxicity.

scholarly journals Expression of myelomonocytic antigens on chronic lymphocytic leukemia B cells correlates with their ability to produce interleukin 1

Blood ◽  
1987 ◽  
Vol 70 (6) ◽  
pp. 1750-1757 ◽  
Author(s):  
F Morabito ◽  
EF Prasthofer ◽  
NE Dunlap ◽  
CE Grossi ◽  
AB Tilden
Keyword(s):  
B Cells ◽  
Chronic Lymphocytic Leukemia ◽  
B Lymphocytes ◽  
Mononuclear Cells ◽  
Gradient Centrifugation ◽  
Interleukin 1 ◽  
Fluorescent Microscopy ◽  
Antigen Expression ◽  
Lymphocytic Leukemia ◽  
Percoll Density Gradient Centrifugation

Abstract We analyzed the expression of myelomonocytic-associated antigens on lymphocytes from B cell chronic lymphocytic leukemia (B-CLL) patients. Blood mononuclear cells were depleted of monocytes by one-step Percoll density gradient centrifugation and tested for antigen expression by fluorescent microscopy and flow cytometry. The reactivity of patient lymphocytes was as follows: 26 of 31 were positive for CD14 (Myr), 22 of 31 for a monocyte Fc receptor (MFC-1), 22 of 31 for CD11b (C3bi receptor), eight of 31 for CD15 (Leu-M1), five of 18 for CD13 (My 7), seven of 18 for My 9, and five of 30 for Mo 2. The B lymphocytes of B- CLL patients were also tested for the ability to produce interleukin 1 (IL-1) after depletion of monocytes and T lymphocytes. In 13 of 17 cases, B lymphocytes of patients produced IL-1 as detected in a mouse thymocyte proliferation assay and, in selected cases, a radioimmunoassay specific for IL-1 beta. The 13 cases that produced IL- 1 were also positive for the expression of one or more myelomonocytic- associated antigens, whereas the four cases that did not produce IL-1 lacked expression of these antigens. In conclusion, the malignant B cells of B-CLL patients frequently express a variety of antigens generally considered specific for myelomonocytic cells, and expression of these antigens is associated with the ability to produce IL-1.

scholarly journals Is Percoll innocuous to cells?

1982 ◽  
Vol 202 (3) ◽  
pp. 795-797 ◽  
Author(s):  
J S Wakefield ◽  
J S J Gale ◽  
M V Berridge ◽  
T W Jordan ◽  
H C Ford
Keyword(s):  
Cell Surface ◽  
Leydig Cells ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Peritoneal Macrophages ◽  
Isolated Rat Liver ◽  
Subcellular Organelles ◽  
Large Numbers ◽  
Gradient Medium ◽  
Isolated Rat

Peritoneal macrophages from mice, isolated rat liver Kupffer cells and rat testis Leydig cells ingested large numbers of Percoll particles, a gradient medium widely used for separation of cells and subcellular organelles by density-gradient centrifugation. A decrease in the percentage of macrophages adhering to plastic also occurred after exposure of the cells to Percoll, even at 4 degrees C, a temperature at which Percoll was not ingested. The effect of Percoll on macrophage adherence may involve a loose association between the density medium and the cell surface. Other cell-surface-related phenomena may also be affected by prior exposure of cells to Percoll.

Rat parietal cell receptors for GLP-1-(7-36) amide: northern blot, cross-linking, and radioligand binding

1994 ◽  
Vol 267 (3) ◽  
pp. G423-G432 ◽  
Author(s):  
J. Schmidtler ◽  
K. Dehne ◽  
H. D. Allescher ◽  
V. Schusdziarra ◽  
M. Classen ◽  
...  
Keyword(s):  
Parietal Cell ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Parietal Cells ◽  
Small Cells ◽  
Cross Linking ◽  
Cell Content ◽  
Chief Cells ◽  
High Affinity ◽  
Isolated Rat

The intestinal peptide hormone glucagon-like peptide-1 (GLP-1) (7-36) amide is a potent stimulus of H+ production in isolated rat parietal cells, suggesting the presence of specific GLP-1-receptors on this cell type. Our aim was to characterize these receptors. Enzymatically isolated rat gastric mucosal cells (F0) were fractionated by counterflow elutriation, resulting in five fractions (F1-F5) according to increasing cell diameter and parietal cell content (3, 5, 4, 27, 81%). Additional density gradient centrifugation of F4 yielded enriched chief cells (74%; parietal cells: 1%; F6), whereas density gradient centrifugation of F5 almost purified parietal cells (97%; chief cells: 1%; F7). Northern blot of total cellular RNA from F0-F7 with a probe specific for the GLP-1-(7-36) amide receptor revealed two RNA species of 2.7 and 3.6 kb. These messages were present to some extent in small cells (F1, F2), much more pronounced in F5, abundant in F7, barely detectable in F3 and F4, and absent from F6. Cross-linking of 125I-labeled GLP-1-(7-36) amide to parietal cell membranes revealed a single 59-kDa band that was abolished by unlabeled GLP-1-(7-36) amide. Throughout fractions F1-F7 specific binding of 125I-GLP-1-(7-36) amide was correlated with parietal cell content (r = 0.99; P < 0.01) and H+ production ([14C]aminopyrine accumulation) in response to GLP-1-(7-36) amide or histamine (r = 0.96; P < 0.01). Binding was maximal in purified parietal cells (F7), whereas almost no binding was detectable in enriched chief cells (F6). In F7, Scatchard analysis revealed a single class of high-affinity binding sites (KD = 2.8 +/- 0.6 x 10(-10) M, Bmax = 6.8 +/- 1.4 fmol/10(6) cells, 4,096 +/- 793 receptors/parietal cells). The following half-maximal inhibition values were found for GLP-1-(7-36) amide and (1-37) and (1-36) amide: 6.6 +/- 0.9 x 10(-10), 1.4 +/- 0.7 x 10(-7), and 2.6 +/- 0.4 x 10(-7) M, respectively. Pancreatic glucagon, GLP-2, and oxyntomodulin, products of the proglucagon gene, were 3-4 log units less potent displacers while gastric inhibitory peptide, vasoactive intestinal peptide, and secretin were ineffective. We conclude that rat parietal cells are equipped with specific high-affinity receptors for GLP-1-(7-36) amide, which, in addition, are present in as yet unidentified small cells (F1, F2) but not in chief cells.

scholarly journals Expression of myelomonocytic antigens on chronic lymphocytic leukemia B cells correlates with their ability to produce interleukin 1

Blood ◽  
1987 ◽  
Vol 70 (6) ◽  
pp. 1750-1757 ◽  
Author(s):  
F Morabito ◽  
EF Prasthofer ◽  
NE Dunlap ◽  
CE Grossi ◽  
AB Tilden
Keyword(s):  
B Cells ◽  
Chronic Lymphocytic Leukemia ◽  
B Lymphocytes ◽  
Mononuclear Cells ◽  
Gradient Centrifugation ◽  
Interleukin 1 ◽  
Fluorescent Microscopy ◽  
Antigen Expression ◽  
Lymphocytic Leukemia ◽  
Percoll Density Gradient Centrifugation

We analyzed the expression of myelomonocytic-associated antigens on lymphocytes from B cell chronic lymphocytic leukemia (B-CLL) patients. Blood mononuclear cells were depleted of monocytes by one-step Percoll density gradient centrifugation and tested for antigen expression by fluorescent microscopy and flow cytometry. The reactivity of patient lymphocytes was as follows: 26 of 31 were positive for CD14 (Myr), 22 of 31 for a monocyte Fc receptor (MFC-1), 22 of 31 for CD11b (C3bi receptor), eight of 31 for CD15 (Leu-M1), five of 18 for CD13 (My 7), seven of 18 for My 9, and five of 30 for Mo 2. The B lymphocytes of B- CLL patients were also tested for the ability to produce interleukin 1 (IL-1) after depletion of monocytes and T lymphocytes. In 13 of 17 cases, B lymphocytes of patients produced IL-1 as detected in a mouse thymocyte proliferation assay and, in selected cases, a radioimmunoassay specific for IL-1 beta. The 13 cases that produced IL- 1 were also positive for the expression of one or more myelomonocytic- associated antigens, whereas the four cases that did not produce IL-1 lacked expression of these antigens. In conclusion, the malignant B cells of B-CLL patients frequently express a variety of antigens generally considered specific for myelomonocytic cells, and expression of these antigens is associated with the ability to produce IL-1.

Protective effect of gap junction uncouplers given during hypoxia against reoxygenation injury in isolated rat hearts

2006 ◽  
Vol 290 (2) ◽  
pp. H648-H656 ◽  
Author(s):  
Antonio Rodríguez-Sinovas ◽  
David García-Dorado ◽  
Marisol Ruiz-Meana ◽  
Jordi Soler-Soler
Keyword(s):  
Protective Effect ◽  
Gap Junction ◽  
Electrical Impedance ◽  
Palmitoleic Acid ◽  
Cell Injury ◽  
Glycyrrhetinic Acid ◽  
Response Analysis ◽  
Rat Hearts ◽  
Ldh Release ◽  
Isolated Rat

It has been shown that cell-to-cell chemical coupling may persist during severe myocardial hypoxia or ischemia. We aimed to analyze the effects of different, chemically unrelated gap junction uncouplers on the progression of ischemic injury in hypoxic myocardium. First, we analyzed the effects of heptanol, 18α-glycyrrhetinic acid, and palmitoleic acid on intracellular Ca2+ concentration during simulated hypoxia (2 mM NaCN) in isolated cardiomyocytes. Next, we analyzed their effects on developed and diastolic tension and electrical impedance in 47 isolated rat hearts submitted to 40 min of hypoxia and reoxygenation. All treatments were applied only during the hypoxic period. Cell injury was determined by lactate dehydrogenase (LDH) release. Heptanol, but not 18α-glycyrrhetinic acid nor palmitoleic acid, attenuated the increase in cytosolic Ca2+ concentration induced by simulated ischemia in cardiomyocytes and delayed rigor development (rigor onset at 7.31 ± 0.71 min in controls vs. 14.76 ± 1.44 in heptanol-treated hearts, P < 0.001) and the onset of the marked changes in electrical impedance (tissue resistivity: 4.02 ± 0.29 vs. 7.75 ± 1.84 min, P = 0.016) in hypoxic rat hearts. LDH release from hypoxic hearts was minimal and was not significantly modified by drugs. However, all gap junction uncouplers, given during hypoxia, attenuated LDH release during subsequent reoxygenation. Dose-response analysis showed that increasing heptanol concentration beyond the level associated with maximal effects on cell coupling resulted in further protection against hypoxic injury. In conclusion, gap junction uncoupling during hypoxia has a protective effect on cell death occurring upon subsequent reoxygenation, and heptanol has, in addition, a marked protective effect independent of its uncoupling actions.

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