scholarly journals Fluorescent methotrexate labeling and flow cytometric analysis of cells containing low levels of dihydrofolate reductase.

1986 ◽  
Vol 261 (14) ◽  
pp. 6285-6292
Author(s):  
P Gaudray ◽  
J Trotter ◽  
G M Wahl
Keyword(s):  
Dihydrofolate Reductase ◽  
Flow Cytometric Analysis ◽  
Flow Cytometric ◽  
Low Levels

Using AFLP markers to inform population management of the endemic Chatham Island toetoe, Austroderia turbaria (Poaceae).

2012 ◽  
Vol 18 (1) ◽  
pp. 33 ◽  
Author(s):  
Gary J Houliston ◽  
Murray I Dawson ◽  
Peter J De Lange ◽  
Peter B Heenan
Keyword(s):  
Management Practices ◽  
Flow Cytometric Analysis ◽  
Natural Populations ◽  
Source Material ◽  
Population Variation ◽  
Population Genetic Analysis ◽  
Flow Cytometric ◽  
Length Polymorphisms ◽  
Amplified Fragment Length Polymorphisms ◽  
Low Levels

Austroderia turbaria Connor is a threatened grass endemic to the Chatham Islands. Although formerly more widespread, remaining natural populations consist of highly fragmented remnants and/or individuals. Population genetic analysis of seed-raised progeny from six of the extant natural populations on Chatham and Pitt islands, using amplified fragment length polymorphisms (AFLPs) and microsatellite markers, shows that there are very low levels of variation (expected estimated heterozygosity He 0.023–0.030, no. of effective alleles Na 1.039–1.053), and no significant differentiation within or between populations on the two islands. Flow cytometric analysis of endosperm to embryo ratios suggests a sexual breeding system. This lack of population variation and no discernable differences between the two islands suggest that management practices such as the establishment of new populations can be carried out irrespective of the location of source material. One caveat to this is the possibility of Fusarium wilt occurring on the islands, in which case measures should be taken to best prevent spread across the range of the species.

Expression of the cell adhesion molecules on leukocytes that demarginate during acute maximal exercise

1999 ◽  
Vol 86 (3) ◽  
pp. 970-976 ◽  
Author(s):  
Stephan F. van Eeden ◽  
John Granton ◽  
Jennifer M. Hards ◽  
Barbara Moore ◽  
James C. Hogg
Keyword(s):  
Fluorescence Intensity ◽  
Mean Fluorescence Intensity ◽  
Maximal Exercise ◽  
Flow Cytometric Analysis ◽  
Cycle Ergometer ◽  
Phenotypic Characteristics ◽  
Flow Cytometric ◽  
The Mean ◽  
Low Levels

The pulmonary vascular bed is an important reservoir for the marginated pool of leukocytes that can be mobilized by exercise or catecholamines. This study was designed to determine the phenotypic characteristics of leukocytes that are mobilized into the circulation during exercise. Twenty healthy volunteers performed incremental exercise to exhaustion [maximal O2consumption (V˙o2 max)] on a cycle ergometer. Blood was collected at baseline, at 3-min intervals during exercise, atV˙o2 max, and 30 min after exercise. Total white cell, polymorphonuclear leukocyte (PMN), and lymphocyte counts increased with exercise toV˙o2 max( P < 0.05). Flow cytometric analysis showed that the mean fluorescence intensity of L-selectin on PMN (from 14.9 ± 1 at baseline to 9.5 ± 1.6 atV˙o2 max, P < 0.05) and lymphocytes (from 11.7 ± 1.2 at baseline to 8 ± 0.8 atV˙o2 max, P < 0.05) decreased with exercise. Mean fluorescence intensity of CD11b on PMN increased with exercise (from 10.2 ± 0.6 at baseline to 25 ± 2.5 atV˙o2 max, P < 0.002) but remained unchanged on lymphocytes. Myeloperoxidase levels in PMN did not change with exercise. In vitro studies showed that neither catecholamines nor plasma collected atV˙o2 maxduring exercise changed leukocyte L-selectin or CD11b levels. We conclude that PMN released from the marginated pool during exercise express low levels of L-selectin and high levels of CD11b.

Virulent and avirulent Entamoeba histolytica and E. dispar differ in their cell surface phosphorylated glycolipids

Parasitology ◽  
1997 ◽  
Vol 114 (2) ◽  
pp. 95-104 ◽  
Author(s):  
S. MOODY ◽  
S. BECKER ◽  
Y. NUCHAMOWITZ ◽  
D. MIRELMAN
Keyword(s):  
Entamoeba Histolytica ◽  
Polyclonal Antibodies ◽  
Virulent Strain ◽  
Flow Cytometric Analysis ◽  
Structural Features ◽  
Surface Distribution ◽  
Flow Cytometric ◽  
Virulent Strains ◽  
Low Levels ◽  
Modified Forms

Virulent strains of Entamoeba histolytica have been reported to produce a mixture of phosphoglycoconjugates that share some structural features with the lipophosphoglycans (LPGs) of Leishmania. Purification of these glycoconjugates is essential to their precise structural characterization. In this study we have extracted ‘LPG-like’ molecules from various virulent E. histolytica strains and purified on the basis of charge differences, 2 apparently related glycoconjugates a ‘LPG’ and a ‘lipophosphopeptidoglycan (LPPG)’. In marked contrast to the abundance of these ‘LPG’ and ‘LPPG’ molecules in the virulent strains, avirulent E. histolytica and E. dispar strains produce either very low, or no detectable levels of LPG, and either low levels or modified forms of ‘LPPG’. Monospecific polyclonal antibodies prepared against that ‘LPG’ of the virulent strain HM-1: IMSS cl6 identified epitopes shared between both the ‘LPG’ and the ‘LPPG’ of this and other virulent strains, using Western blot analysis. Flow cytometric analysis of a range of strains using these antibodies identified a surface distribution of these molecules and confirmed a correlation between surface exposure of epitopes bound by these antibodies and parasite virulence.

Apoptotic Effects of Melittin on 4T1 Breast Cancer Cell Line is associated with Up Regulation of Mfn1 and Drp1 mRNA Expression

2020 ◽  
Vol 20 (7) ◽  
pp. 790-799 ◽  
Author(s):  
Farnaz D. Moghaddam ◽  
Pejman Mortazavi ◽  
Somayeh Hamedi ◽  
Mohammad Nabiuni ◽  
Nasim H. Roodbari
Keyword(s):  
Breast Cancer ◽  
Mrna Expression ◽  
Hemolytic Activity ◽  
Breast Cancer Cells ◽  
Flow Cytometric Analysis ◽  
Control Group ◽  
Flow Cytometric ◽  
4T1 Cells ◽  
4T1 Breast Cancer ◽  
Apoptotic Effects

Background and Purpose: Melittin, as the main ingredient of honeybee venom, that has shown anticancer properties. The present study aimed at investigating the cytotoxic impacts of melittin on 4T1 breast cancer cells. Methods: Hemolytic activity of different concentrations (0.125, 0.25, 0.5, 1, 2, 4, 8μg/ml) of melittin was assayed and then cytotoxicity of selected concentrations of melittin (2, 4, 8, 16, 32, and 64μg/ml), 2 and 4μg/ml of cisplatin and 0.513, 0.295 and 0.123μg/ml of doxorubicin was evaluated on 4T1 cells using MTT assay. We used Morphological evaluation and flow cytometric analysis was used. Real time PCR was also used to determine mRNA expression of Mfn1 and Drp1 genes. Results: All compounds showed anti-proliferative effects on the tumor cell line with different potencies. Melittin had higher cytotoxicity against 4T1 breast cancer cells (IC50= 32μg/ml-72h) and higher hemolytic activity (HD50= 1μg/ml), as compared to cisplatin and doxorubicin. Mellitin at 16 and 32μg/ml showed apoptotic effects on 4T1 cells according to the flow cytometric analysis. The Real time PCR analysis of Drp1 and Mfn1 expression in cells treated with 16μg/ml of melittin revealed an up-regulation in Drp1 and Mfn1 genes mRNA expression in comparison with control group. Treatment with 32μg/ml of melittin was also associated with a rise in mRNA expression of Drp1 and Mfn1 as compared to the control group. Conclusion: The results of this study showed that melittin has anticancer effects on 4T1 cell lines in a dose and time dependent manner and can be a good candidate for further research on breast cancer treatment.

scholarly journals Flow cytometric analysis of T lymphocytes and cytokines in aqueous humor of patients with varicella zoster virus-mediated acute retinal necrosis

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Hao Kang ◽  
Yunbo Wei ◽  
Ming Liu ◽  
Di Yu ◽  
Yong Tao
Keyword(s):  
T Lymphocytes ◽  
Varicella Zoster Virus ◽  
Aqueous Humor ◽  
T Lymphocyte ◽  
Lymphocyte Subsets ◽  
Flow Cytometric Analysis ◽  
T Lymphocyte Subsets ◽  
Varicella Zoster ◽  
Flow Cytometric ◽  
Cd8 T Lymphocytes

Abstract Background The purpose of this study is to investigate the aqueous humor (AH) T lymphocyte subsets and cytokines of acute retinal necrosis (ARN) to elucidate the immunologic inflammatory features of this disorder. Methods Three patients with ARN infected with varicella zoster virus (VZV) who underwent multiple intravitreal injections of ganciclovir were enrolled in this study. The control group consisted of four non-infectious patients with acute anterior uveitis (AAU). Flow cytometric analysis was performed on the lymphocyte subsets from the AH and peripheral blood (PB) samples during the active phase of intraocular inflammation. Five inflammatory cytokines were measured in each AH sample and various clinical characteristics were also assessed. Results VZV deoxyribonucleic acid (DNA) was detected by real-time polymerase chain reaction (PCR) in AH from all the ARN patients, who showed higher CD8+ T lymphocytes population in AH than the AAU patients (p = 0.006). CD4/CD8 ratios of T lymphocytes and the percentage of CD8 + CD25+ T lymphocytes in AH were significantly lower in ARN than in AAU (p = 0.006; p = 0.012). In the ARN patients, the percentages of CD4+ and CD8+ T lymphocytes in AH were higher than those found in PB. The percentage of CD4 + CD25+ T lymphocytes in AH was significantly higher than the proportion in PB in the AAU patients (p = 0.001). Immunoregulatory cytokine Interleukin-10 in AH was significantly elevated in the ARN patients in comparison with the case of the AAU patients (p = 0.036). In ARN, the copy number of VZV DNA in AH positively correlated with the percentage of CD8+ T lymphocytes in AH and negatively correlated with the CD4/CD8 ratio in AH during the course of disease treatment (p = 0.009, r = 0.92; p = 0.039, r = − 0.834). Conclusion The ARN patients caused by VZV had different intraocular T lymphocyte subsets and cytokines profile than those of the non-infectious patients. High percentages of CD8+ T lymphocytes and low CD4/CD8 T cell ratios may be a potential biomarker for diagnosis of viral-infectious uveitis. T lymphocytes examination at the inflammatory sites has the potential to become a useful research tool for differentiating viral and non-viral uveitis.

scholarly journals LncRNA NEAT1 Enhances Glioma Progression via Regulating the miR-128-3p/ITGA5 Axis

2021 ◽  
Author(s):  
Jiakai Chen ◽  
Handong Wang ◽  
Junjun Wang ◽  
Wenhao Niu ◽  
Chulei Deng ◽  
...  
Keyword(s):  
Flow Cytometric Analysis ◽  
Biological Functions ◽  
Quantitative Real Time Pcr ◽  
Flow Cytometric ◽  
Non Coding Rna ◽  
Rna Immunoprecipitation ◽  
Human Gbm ◽  
Novel Strategy ◽  
Lncrna Neat1 ◽  
Long Non Coding Rna

AbstractAccumulating evidences indicate that long non-coding RNA nuclear paraspeckle assembly transcript 1 (NEAT1) promotes the progression of glioma. In this study, we postulated that NEAT1 may act as a miR-128-3p sponge. Relative levels of NEAT1 and miR-128-3p expression in human glioma samples and GBM cells were detected using quantitative real-time PCR. By means of CCK-8 assays, transwell assays, and flow cytometric analysis, the biological functions of miR-128-3p and NEAT1 were investigated in U87MG and U251MG human GBM cell lines with stable miR-128-3p and NEAT1 knockdown or overexpression. The luciferase reports, RNA pull-down assay, and RNA immunoprecipitation assay were conducted to determine the relevance of NEAT1 and miR-128-3p in glioma. As a result, high expression of NEAT1 and lack of miR-128-3p were observed in glioma specimens and cells. By binding to anti-oncogene miR-128-3p in the nucleus, NEAT1 enhanced tumorigenesis and glioma development. Further experiments suggested that ITGA5 expression was increased in glioma tissues and was found to be connected with miR-128-3p. Additionally, NEAT1 facilitated ITGA5 expression via competitively binding to miR-128-3p. For this reason, ITGA5 would not be decomposed by miR-128-3p and could activate FAK signaling pathway, thereby promoting cell growth. Collectively, these results indicated that the NEAT1/miR-128-3p/ITGA5 axis was involved in glioma initiation and progression, and might offer a potential novel strategy for treatment of glioma.

Sign in / Sign up

Export Citation Format

Share Document