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1.
Article in English | MEDLINE | ID: mdl-26464574

ABSTRACT

BACKGROUND: Venoms comprise mixtures of numerous bioactive compounds that have a wide range of pharmacologic actions. Toxins from venomous animals have attracted the attention of researchers because of their affinity for primary sites responsible for lethality and their efficacy at extremely low concentrations. The venoms of marine stingrays have not been extensively studied and limited data is available on them. The present study aims to evaluate the antiproliferative and biochemical properties of the venom obtained from a species of marine stingray (Dasyatis sephen) on human cervical cancer cell line HeLa. METHODS: The antiproliferative effect of D. sephen venom was determined by MTT assay, and the oxidative stress was determined by lipid peroxidation method along with assessment of changes in the enzymatic and non-enzymatic antioxidant status. We observed intracellular reactive oxygen species (ROS) levels by DCFH-DA method, mitochondrial membrane potential alterations by rhodamine 123 staining and apoptotic morphological changes by acridine orange/ethidium bromide dual staining method. RESULTS: D. sephen venom enhances lipid peroxidative markers such as thiobarbituric acid reactive substance, conjugated diene, and lipid hydroperoxide in HeLa cell lines. Stingray venom enhances the ROS levels, which is evidenced by the increased 2-7-diacetyl dichlorofluorescein fluorescence. Further, D. sephen venom treatment altered the mitochondrial membrane potential in HeLa cells. Additionally, we observed increased apoptotic morphological changes in D. sephen venom-treated groups. CONCLUSIONS: Dasyatis sephen venom exhibits potent antiproliferative effect on HeLa cell line and upon further purification it could be a promising antiproliferative agent.

2.
J. venom. anim. toxins incl. trop. dis ; 21: 1-9, 31/03/2015. ilus, graf
Article in English | LILACS, VETINDEX | ID: biblio-1484637

ABSTRACT

Background Venoms comprise mixtures of numerous bioactive compounds that have a wide range of pharmacologic actions. Toxins from venomous animals have attracted the attention of researchers because of their affinity for primary sites responsible for lethality and their efficacy at extremely low concentrations. The venoms of marine stingrays have not been extensively studied and limited data is available on them. The present study aims to evaluate the antiproliferative and biochemical properties of the venom obtained from a species of marine stingray (Dasyatis sephen) on human cervical cancer cell line HeLa.MethodsThe antiproliferative effect of D. sephen venom was determined by MTT assay, and the oxidative stress was determined by lipid peroxidation method along with assessment of changes in the enzymatic and non-enzymatic antioxidant status. We observed intracellular reactive oxygen species (ROS) levels by DCFH-DA method, mitochondrial membrane potential alterations by rhodamine 123 staining and apoptotic morphological changes by acridine orange/ethidium bromide dual staining method.ResultsD. sephen venom enhances lipid peroxidative markers such as thiobarbituric acid reactive substance, conjugated diene, and lipid hydroperoxide in HeLa cell lines. Stingray venom enhances the ROS levels, which is evidenced by the increased 27-diacetyl dichlorofluorescein fluorescence. Further, D. sephen venom treatment altered the mitochondrial membrane potential in HeLa cells. Additionally, we observed increased apoptotic morphological changes in D. sephen venom-treated groups. ConclusionsDasyatis sephen venom exhibits potent antiproliferative effect on HeLa cell line and upon further purification it could be a promising antiproliferative agent.


Subject(s)
Humans , Animals , Animals, Poisonous , Venoms/therapeutic use , Skates, Fish , HeLa Cells/drug effects
3.
J. venom. anim. toxins incl. trop. dis ; 21: 41, 31/03/2015. graf, ilus
Article in English | LILACS, VETINDEX | ID: biblio-954762

ABSTRACT

BackgroundVenoms comprise mixtures of numerous bioactive compounds that have a wide range of pharmacologic actions. Toxins from venomous animals have attracted the attention of researchers because of their affinity for primary sites responsible for lethality and their efficacy at extremely low concentrations. The venoms of marine stingrays have not been extensively studied and limited data is available on them. The present study aims to evaluate the antiproliferative and biochemical properties of the venom obtained from a species of marine stingray (Dasyatis sephen) on human cervical cancer cell line HeLa.MethodsThe antiproliferative effect of D. sephen venom was determined by MTT assay, and the oxidative stress was determined by lipid peroxidation method along with assessment of changes in the enzymatic and non-enzymatic antioxidant status. We observed intracellular reactive oxygen species (ROS) levels by DCFH-DA method, mitochondrial membrane potential alterations by rhodamine 123 staining and apoptotic morphological changes by acridine orange/ethidium bromide dual staining method.ResultsD. sephen venom enhances lipid peroxidative markers such as thiobarbituric acid reactive substance, conjugated diene, and lipid hydroperoxide in HeLa cell lines. Stingray venom enhances the ROS levels, which is evidenced by the increased 2-7-diacetyl dichlorofluorescein fluorescence. Further, D. sephen venom treatment altered the mitochondrial membrane potential in HeLa cells. Additionally, we observed increased apoptotic morphological changes in D. sephen venom-treated groups. ConclusionsDasyatis sephen venom exhibits potent antiproliferative effect on HeLa cell line and upon further purification it could be a promising antiproliferative agent.(AU)


Subject(s)
Animals , Skates, Fish , Carcinoma , Oxidative Stress , Cell Line
4.
Methods Enzymol ; 509: 165-94, 2012.
Article in English | MEDLINE | ID: mdl-22568906

ABSTRACT

Design of a unique, single-platform, integrated, multichannel sensor based on carbon nanotube (CNT)-protein adducts specific to each one of the major analytes of blood, glucose, cholesterol, triglyceride, and Hb1AC is presented. The concept underlying the sensor, amperometric detection, is applicable to various disease-monitoring strategies. There is an urgent need to enhance the sensitivity of glucometers to <5% level instead of greater than the present 15% standard in these detectors. CNTs enhance the signals derived from the interaction of the enzymes with the different analytes in blood. Fabricated sensors using the new methodology is a point-of-care device that is targeted for home, clinical, and emergency use and can be redesigned for continuous monitoring for critical care patients.


Subject(s)
Biosensing Techniques , Blood Chemical Analysis/methods , Nanotubes, Carbon , Amino Acid Oxidoreductases/biosynthesis , Amino Acid Oxidoreductases/chemistry , Blood Glucose/analysis , Cholesterol/blood , Cholesterol Oxidase/biosynthesis , Cholesterol Oxidase/chemistry , Glucose Oxidase/biosynthesis , Glucose Oxidase/chemistry , Glycated Hemoglobin/metabolism , Humans , Immobilized Proteins/biosynthesis , Immobilized Proteins/chemistry , Lab-On-A-Chip Devices , Lipase/biosynthesis , Lipase/chemistry , Microfluidics , Models, Molecular , Protein Engineering , Protein Structure, Tertiary , Recombinant Proteins/biosynthesis , Recombinant Proteins/chemistry , Sterol Esterase/biosynthesis , Sterol Esterase/chemistry , Surface Properties , Triglycerides/blood
5.
Asian Pac J Allergy Immunol ; 14(2): 87-90, 1996 Dec.
Article in English | MEDLINE | ID: mdl-9177821

ABSTRACT

The aim of this project was to compare dual and tri-colour reagents for lymphocyte immunophenotyping. A total of 37 patient and normal specimens were immunophenotyped concurrently with the following mean values (% dual vs tri-colour): CD3 (69.4 vs 68.3) CD4 (24.0 vs 24.2) and CD19 (13.9 vs 12.6). A comparison of the results obtained using the paired t test showed that there were no significant differences for cells expressing CD3, CD4 and CD19. However, there was a significant difference in the NK (18.3 vs 16.3) cell component. A major advantage in using 3 colour immunophenotyping is the ability to analyse specimens that cannot be analysed using dual colour reagents due to debris or contamination of the gate with non-lymphocytic cells.


Subject(s)
Coloring Agents , Indicators and Reagents , Lymphocyte Subsets/cytology , Antigens, CD19/analysis , CD3 Complex/analysis , CD4 Antigens/analysis , Flow Cytometry/methods , HLA-DR Antigens/analysis , Humans , Killer Cells, Natural/chemistry , Killer Cells, Natural/cytology , Lymphocyte Subsets/chemistry
6.
Ann Acad Med Singap ; 24(6): 785-8, 1995 Nov.
Article in English | MEDLINE | ID: mdl-8838981

ABSTRACT

A cross-sectional study on the expression of 6 lymphocyte markers was carried out on 481 patients with human immunodeficiency virus (HIV) and 79 normals after stratification based on absolute CD4 counts. The data were stratified according to the following groups: (I) 1201 to 1600, (II) 801 to 1200, (III) 401 to 800 and (IV) 0 to 400 (x 10(6) CD4 cells per mm3). The mean percentages of the subsets before stratification showed that HIV patients had increased percentages of CD3+ (75.7 against 66.9), CD3+CD8+ (52.2 against 32.3) and CD3+HLA-DR+ (36.1 against 14.4) cells and lower percentages of CD19 (10.3 against 13.3) and natural killer cells (13.7 against 20.4) when compared to controls in the same group. A definite trend, however, was only seen in CD3+CD8+ (47.4, 50.0, 54.0, 57.5 for groups I, II, III and IV respectively) and CD3+HLA-DR+ (29.1, 32.9, 38.4, 43.9 for groups I, II, III and IV respectively).


Subject(s)
CD4 Lymphocyte Count , CD4-Positive T-Lymphocytes/pathology , HIV Seropositivity/immunology , Lymphocyte Activation/immunology , T-Lymphocytes/immunology , Antigens, CD19/genetics , B-Lymphocytes/pathology , CD3 Complex/genetics , CD8 Antigens/genetics , CD8-Positive T-Lymphocytes/pathology , Cross-Sectional Studies , Gene Expression Regulation , HIV Seropositivity/pathology , HLA-DR Antigens/genetics , Humans , Killer Cells, Natural/pathology , Lymphocyte Count , Retrospective Studies , T-Lymphocytes/pathology , T-Lymphocytes, Helper-Inducer/pathology
7.
Singapore Med J ; 36(3): 288-91, 1995 Jun.
Article in English | MEDLINE | ID: mdl-8553095

ABSTRACT

The aim of this study was to establish the lymphocyte subset reference ranges in a defined Malaysian population as well as to determine inter-racial differences for these values. Normal blood obtained from 152 subjects (55.9% Malay, 26.3% Chinese and 17.7% Indian) was immunophenotyped. Results obtained (expressed as mean +/- SD %), absolute count (x 10(6) cells/mm3) were as follows: CD3:66.5 +/- 8.6%, 2,066; CD4:33.2 +/- 8.5%, 1,028; CD831.6 +/- 8.9%, 982; CD19:12.0 +/- 0%, 5,374, and CD56+CD16:20.9 +/- 9%, 1,638. There were no significant differences between the percent lymphocyte subsets of the three racial groups. However, the absolute number of CD4 cells and CD19 cells in Chinese was significantly lower (p < 0.05) compared to the Indian and the Indian and Malay groups respectively. Comparison of our results with other reports showed that the percentage of Natural Killer cells in this population is higher than that reported for Caucasian population.


Subject(s)
Asian People , Lymphocyte Subsets/chemistry , White People , Adolescent , Adult , China/ethnology , Female , Humans , Immunophenotyping , India/ethnology , Malaysia , Male , Middle Aged , Reference Values
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