The effect of excessive disodium ethylene diamine tetraacetic acid (Na2EDTA) anticoagulant concentration on leukocytes profile in peripheral blood examination.

Accuracy of leukocytes profile assessment is influenced by several pre analytical factors, among others, the anticoagulant concentration. EDTA is one of the most frequently used anticoagulant in peripheral blood examination. Several references stated that inappropriate concentration of EDTA anticoagulant in blood sample may affect the result of leukocytes profile in peripheral blood examination. The aim of this study was to evaluate whether there are differences among leukocytes profile in peripheral blood examination specimens, which were prepared with excessive Na2EDTA anticoagulant in different concentration. This study was conducted in Faculty of Medicine, Gadjah Mada University. Blood samples from 30 subjects were taken using vein puncture. Two millimeters blood was divided into 4 Na2EDTA-containing tubes. Before that, one drop of blood without Na2EDTA anticoagulant was used to make blood film right after vein puncture, as control. Each tubes contained different concentration of anticoagulant. The first tube contained Na2EDTA in standard concentration 2 mg/ml; the remaining tubes contained 4 mg/ml, 6 mg/ml, and 8 mg/ml respectively. These samples were immediately examined using SYSMEX SE-9500 automatic cell counter to measure the total and differential leukocytes count; and were stained with Wright staining for morphological examination under the microscope. These procedures were done before 20 minutes of vein puncture. There were significant decrement of total leukocytes count, absolute differential leukocytes count and monocyte percentage following excessive Na2EDTA administration. Neutrophil percentage was found to be relatively increased and the difference was significant. Lymphocyte, eosinophil and basophil percentages were not significantly different. Morphological examination showed significant increment in irregular cytoplasm margin, vacoulation and irregular nuclei lobes following excessive Na2EDTA administration. It is concluded that excessive concentration of Na2EDTA used in blood specimen preparation, will lead to changes in leukocytes profile as the concentration increased. Standard Na2EDTA anticoagulant concentration did not alter any leukocytes count and morphology, except for irregular cytoplasm margin and irregular nuclei lobes.

The effects of excessive Disodium Ethylene Diamine Tetraacetic Acid (Na2EDTA) anticoagulant concentration toward hematology profile and morphology of erythrocytes in peripheral blood examination.

The purpose of this study is to know whether there are differences between hematology profile and morphology of erythrocytes of blood specimens which are prepared with excessive Na2EDTA anticoagulant in different concentration. This study was conducted in Faculty of Medicine Gadjah Mada University. The criteria of subject were male, age from 18 until 22 years old and healthy, ascertained from history taking and vital sign examination. Blood samples from 33 subjects were taken using vein puncture. Two millimeters blood was divided into 4 Na2EDTA-containing tubes. Before that, one drop of blood without Na2EDTA anticoagulant was used for making control blood film right after vein puncture. Each tubes contained different concentration of anticoagulant. The first tube contained Na2EDTA in standard concentration 2 mg/dl; the remaining tubes contained consecutively, 4 mg/dl, 6 mg/dl, and 8 mg/dl. Those samples were immediately examined using SYSMEX SE-9500 automatic analyzer for measuring erythrocytes hematological profile and were stained with Wright staining for morphological examination. These procedures were done before 20 minutes of vein puncture. There were significant decrease of RBC count, HGB, HCT, and MCHC and also significant increase of MCV and RDW between different concentrations of excessive Na2EDTA anticoagulant. MCH did not have significant result. Morphological examination showed significant morphological changes in the form of echinocytes and appearance of ghost cells in the sample treated with excessive Na2EDTA anticoagulant concentration. In conclusion, there are differences in hematological profile and morphology of erythrocytes among blood specimen which are prepared with excessive Na2EDTA anticoagulant in different concentration, except for MCH. Excessive Na2EDTA anticoagulant concentration will affect the blood specimen for peripheral blood examination of erythrocytes by interfering morphology and some of hematological parameters.

Polyphenols extracted from the Green Tea (Camellia sinensis) augments the protective immune responses in mice challanged with Salmonella typhimurium.

Green tea is an aqueous infusion of dried unfermented leaves of Camellia sinensis. Numerous biological activities of green tea have been reported. The aqueous infusion and its polyphenolic substance are known for their activity as an antimutagenic, antibacterial, hypocholesterolemic, antioxidant, and mutagenic of B lymphocyte. Studies have demonstrated that green tea polyphenols increase IL-12 production. Salmonella spp infection is an important public health problem in many countries. Cell-mediated immunity (CMI), especially T-cell help is important for protection against this infection. Recent evidence indicates that IL-12 is one such factor that plays a crucial role in the development of CMI. These studies were carired out to investigate the effect of green tea polyphenols to the immune cellulare in mice responses of mice during Salmonella typhimurium infection. The subject consisted of 36 female mice (Balb/C), 6-8 weeks old, divided into 3 groups. The first group was given 10 mg polyphenols/mouse, the second group was given 5 mg polyphenols/mouse, and the third group as the control. In day 31, all mice were infected with 108 CFU Salmonella typhimurium orally. On day 0, 3, 5, and 7 postinfection, 3 mice from each groups were sacrificed, the splenocytes were extracted and cultured to measure the level of IFN-g in the supernatan and. The peritoneal macrophages were also extracted and cultured to measure the phagocytic activity. The level of IFN-g in splenocyte culture supernatant increased during infection in all groups, but the level of the experimental groups were higher than in control group. The percentage of phagocytic activity of peritoneal macrophages were higher in the experimental groups than in the control group. The increase of the phagocytic activities were seen corelate with the level of IFN-g supernatan splenocyte culture.