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P Butthep

Publications and source records attributed to P Butthep.

At least 19 recordsLinked to original sources
PubMed2006

Cytoadherence between endothelial cells and P. falciparum infected and noninfected normal and thalassemic red blood cells.

BACKGROUND: Cytoadhesion of P. falciparum infected red blood cells (RBCs) to endothelial cells (ECs) is an important phenomenon that causes cerebral malaria in man. Reduced adhesion especially in thalassemia and hemoglobinopathies may be related to a protective mechanism against malaria in such people. METHODS: The cytoadherence assay was performed using both conventional and floating conditions between ECs (ECV 304) and P. falciparum infected and noninfected RBCs from both normal and thalassemia subjects. In floating condition, RBC was fluorescently labeled with anti-glycophorin A antibody, whereas EC was identified by surface expression of PECAM-1, CD-36, ICAM-1, and E-selectin. The condition of floating EC was similar to the condition for subcultivation as they can adhere or bind to any surface. The phosphatidylserine (PS) exposure was also determined by using flow cytometer. RESULTS: The adhesion of noninfected heterozygous thalassemic RBCs (all genotypes) to ECs was significantly increased as compared with normal RBCs (P < 0.02). Interestingly, after P. falciparum infection, the number of normal RBCs bound to ECs was significantly increased as compared with noninfected RBCs (P < 0.01), whereas heterozygous thalassemic RBCs infected by P. falciparum showed no significant difference compared with noninfected RBCs. In addition, we found a similar level of PS exposure in normal and thalassemic infected RBCs, which was related to the cytoadherence phenomenon. CONCLUSION: The reduced adhesion between heterozygous thalassemic RBCs infected by P. falciparum to ECs provides an explanation for their protective mechanism against malaria, as increased adhesion is a high risk for cerebral malaria and nonbinding infected RBCs can be removed by the reticuloendothelial system and other mechanism(s) in vivo.

Animals↗
PubMed2000

Enhanced tumor necrosis factor in the serum and renal hypoperfusion in nephrosis associated with focal segmental glomerulosclerosis.

Enhanced tumor necrosis factor alpha associated with immunocirculatory imbalance expressed as increased ratio between proinflammatory (TNFalpha) and antiinflammatory (IL-10) cytokines was observed in the serum of nephrosis associated with focal segmental glomerulosclerosis. Such altered immunocirculatory balance correlated with the reduction in renal plasma flow determined by the intrarenal hemodynamic study by which it implies that a glomerular endothelial cell injury associated with impaired renal perfusion is likely to be spontaneously induced by enhanced tumor necrosis factor in the presence of inadequate release of antiinflammatory cytokine (IL-10).

Biomarkers↗
PubMed2000

Reticulocyte analysis in iron deficiency anemia and hemolytic anemia.

Reticulocyte analysis was studied in 28 anemic patients, 15 with iron deficiency anemia (IDA), and 13 with hemolytic anemia including 9 glucose 6 phosphate dehydrogenase deficiency (G6PD def.), and 4 with G6PD def. combined with HbE trait or alpha thalassemia trait (alpha thal trait). The reticulocyte analysis among these patients showed the increased number of reticulocyte percentage with low degree of maturation in both IDA and G6PD def. patients. The significantly decreased reticulocyte hemoglobin content (CHr) was found in IDA (CHr = 21.74 +/- 4.61 pg in IDA vs 28.41 +/- 1.34 pg in normal; p-value = < 0.0001), whereas, increased CHr was found in G6PD def. patients. In addition, the G6PD def. patients also showed a significant increase in mean corpuscular reticulocyte volume (MCVr) when compared to normal (MCVr = 132.0 +/- 8.39 fl. in G6PD def. vs 110.39 +/- 5.09 in normal; p-value = < 0.0001). However, a significant decrease in MCVr was found in IDA patients (MCVr = 95.89 +/- 8.57 fl.; p-value = < 0.0001 vs normal). From this study, we can suggest that the reticulocyte hemoglobin content (CHr) and mean corpuscular reticulocyte volume (MCVr) are the important defects in patients with iron deficiency anemia.

Analysis of Variance↗
PubMed2000

Reticulocyte analysis in systemic lupus erythematosus and chronic renal failure using flow cytometry.

The number and maturation of circulating reticulocytes were measured in patients with systemic lupus erythematosus (SLE) and chronic renal failure (CRF) using an automated hematological analyzer (Technicon H*3 RTX) for their erythropoietic activities. Both SLE and CRF patients had increased reticulocyte numbers with a low degree of maturation. The SLE patients had no changes in mean reticulocyte corpuscular volume (MCVr) as compared to normal subjects (110.20 +/- 15.43 fl. in SLE and 110.39 +/- 5.09 fl. in normal), whereas CRF patients had significantly increased mean corpuscular reticulocyte volume (MCVr = 120.99 +/- 8.09 fl., p-value = 0.0019 as compared with normal). Three cases of SLE with nephrotic syndrome (NS) had high degree of MCVr (113.4, 125.0 and 133.1 fl., respectively). The renal involvement in SLE patients and CRF patients may associate with increased reticulocyte corpuscular volume.

Anemia↗
PubMed1997

Dengue viruses induce cell proliferation and morphological changes of endothelial cells.

Replication of dengue viruses (type 1, 2, 3 and 4) in vitro in endothelial cells from human umbilical cord vein was demonstrated by virus titers and immunofluorescent antibody studies. Both showed highest peak at Day 6 after inoculation and declined to origin at Day 14. Some of the cultured endothelial cells detached from the culture well. Most of these floating cells were rarely viable as shown by failure in trypan blue exclusion whereas the adhering cells are mostly viable. More frequent and higher intensity of immunofluorescent positive cells were found in the detached cells as compared to adhering cells. The virus titers in the supernatant and in the adhering cell population were comparable, although floating cells were maximally 26.2% of the total cultured endothelial cells. Many floating cells and occasional adhering cells had numerous blebs on their surface. Endothelial cell proliferation was markedly increased after virus inoculation as compared with the control. Increased number of mitotic cells was also observed in the dengue virus-endothelial cell culture. Comparing among the four types, dengue type 4 induced highest peaks of cell proliferation and cell mitosis at Day 10 after inoculation. Dengue type 2 had the highest virus titers both in adhering cells and in supernatant at Day 6 as compared with other types.

Cell Division↗
PubMed1997

Malaria protection in hereditary ovalocytosis: relation to red cell deformability, red cell parameters and degree of ovalocytosis.

In the culture of red cells with Plasmodium falciparum, erythrocytes from both Thai patients and subjects (patient's parents) with hereditary ovalocytosis have a protective effect against malarial infection. High percentage of ovalocyte (75-100%) was found in patients whereas their parents had lower percentage (25-50%). Invasion index (II) and multiplication ratio (MR) of P. falciparum in these abnormal red cells from the patients were significantly decreased as compared to those in normal red cells (patients: II = 1.52 +/- 0.91, MR = 8.83 +/- 6.73; normal subjects: II = 4.45 +/- 1.51, MR = 25.23 +/- 6.25). This suggests that the red cells from these patients had significant degree of malaria protection. The significant protection was also shown in red cells from the parent group (II = 1.86 +/- 0.81, MR = 15.69 +/- 3.50). Although the parents had lower ovalocyte percentage, degree of protection against malaria parasite was as effective as those found in patients with high ovalocytic red cells. This has been confirmed by statistical analysis showing nonsignificant difference in II value between the two groups. In contrast, red cells of both groups had poor deformability (deformability index, DI) as compared to the normal group. No statistically different DI values were demonstrated between the two. This indicates that poorly deformable red cells, not their ovalocytic shape, make a significant contribution to limitation of malaria parasite invasion. The MR values in patients were less than those found in the parent group but statistical analysis showed no significant difference. Reduced MR values were found with increased numbers of microcytic, hyperchromic and hypochromic red cells in patients.

Elliptocytosis, Hereditary↗
PubMed1997

Possible evidence of endothelial cell activation and disturbance in thalassemia: an in vitro study.

Activation of vascular endothelium is considered as an important facet of inflammation, thrombosis, and vasculitis. Activated endothelial cells express a number of immunologically relevant surface markers which are not detected in dormant condition. These surface markers on endothelial cell may involve in adhesion reaction and migration of blood cell components. We demonstrated increased level of the soluble adhesion molecules in circulating blood of both alpha- and beta-thalassemic patients. These adhesion molecules are theoretically known to be released from endothelial cells. The adhesion molecules included soluble Intercellular Adhesion Molecule-1 (sICAM-1), soluble E-Selectin (ELAM-1), soluble Vascular Cell Adhesion Molecule-1 (sVCAM-1), and von Willebrand Factor (vWF). The levels of these adhesion molecules were measured in serum from 32 thalassemic patients and 10 control healthy subjects. As compared to normal, increased sICAM-1 was found in beta-thal/HbE patients with non-splenectomy; BE-NS (p = 0.002), increased ELAM-1 in beta-thal/HbE patients with splenectomy; BE-S (p = 0.01) and HbH with Hb Constant Spring; HbH/CS (p = 0.001), and increased sVCAM-1 in BE-NS; (p = < 0.0001) and BE-S (p = 0.002). Significant increase in von Willebrand Factor (vWF), a marker for endothelial cell, was shown in BE-S (p = 0.04) as compared to normal. Adhesion molecules were also markedly demonstrated in the supernatant of in vitro culture of human vascular endothelial cell in the presence of 30% thalassemic serum, and these adhesion molecules were also detected on the surface of the cells by using the technic of laser scanning confocal microscope and direct immunofluorescence.

Adult↗
PubMed1997

Thalassemic serum inhibits endothelial cell mitosis in vitro.

Human umbilical vein endothelial cells were cultured in vitro using Iscove's Modified Dulbecco's Medium (IMDM) supplemented with either pooled normal human serum, or pooled thalassemic serum, or autologous umbilical cord serum, or fetal bovine serum. The mitotic activity was determined under the inverted phase contrast microscope and the number of mitotic cells was counted. Our results showed that the mitotic cells decreased in endothelial cell culture with thalassemic serum as compared with normal human serum, autologous umbilical cord serum or fetal bovine serum. The percentage of mitotic cells decreased on day 3 in the presence of beta-thalassemia/HbE serum from both splenectomized and non-splenectomized patients as compared with normal or autologous umbilical cord serum. In the presence of alpha-thalassemic serum, a similar outcome was also observed. From this study we can conclude that the thalassemic sera might contain factors which affect the endothelial cell growth and proliferation by inhibiting mitosis in vitro.

Cells, Cultured↗
PubMed1997

Surface and total tissue factor activity of endothelial cells.

With a technic that was developed by us, we found that normal human umbilical vein endothelial cells (HUVEC) in culture characteristically had very little tissue factor (TF) activity either on the surface or in the cells which had been disrupted. In the presence of endotoxin (E. coli O26:B6), a trigger for thrombosis in septicemic patients, we could not detect an increased TF activity of HUVEC on its surface. However, an increase in TF (total TF) was detected after disruption of the cells. The increase in total TF was dose-dependent. Endotoxin at the concentration of 10 micrograms/ml caused around 5 fold increase in total TF activity compared to that of HUVEC in the absence of endotoxin.

Cells, Cultured↗
PubMed1995

Alterations in vascular endothelial cell-related plasma proteins in thalassaemic patients and their correlation with clinical symptoms.

An increased level of plasma thrombomodulin (TM) in alpha- and beta-thalassaemia was demonstrated using an enzyme-linked immunosorbent assay (ELISA). Nonsplenectomized patients with beta-thalassaemia/haemoglobin E (BE) had higher levels of TM than splenectomized cases (BE-S). Patients with leg ulcers (BE-LU) were found to have the highest increase in TM level. Appearance of larger platelets in all types of thalassaemic blood was observed indicating an increase in the number of younger platelets. These data indicate that injury of vascular endothelial cells is present in thalassaemic patients.

Adolescent↗
PubMed1995

Alterations and pathology of thalassemic red cells: comparison between alpha- and beta-thalassemia.

Two main types of thalassemia have been categorized according to defective production of the globin gene ie alpha-thalassemia and beta-thalassemia. We report different red cell abnormalities between these two types. The study included 139 thalassemic patients including 91 patients with hemoglobin (Hb) H disease (52 cases with the classical genotype and 39 cases with Hb Constant Spring) and 48 were beta-thalassemia/Hb E disease. The deformability index of thalassemic red cells measured by laser diffractometer was significantly lower than that of normal red cells. Increased susceptibility of the thalassemic red cells to monocyte phagocytosis was markedly noted. Few sialic acid molecules were scattered on red cell surface of thalassemic red cells. Reticulocytes with delayed maturation stage were also observed in thalassemia indicating enhanced release from the bone marrow. The alpha-thalassemic red cells had relatively better deformability, increased susceptibility to phagocytosis, reduced sialic acid content and greater degree irregular distribution of sialic acid on red cell surface as compared to beta-thalassemic red cells. The alpha-type with hemoglobin Constant Spring (Hb CS) had increased percentage of reticulocyte and young reticulocyte (high fluorescent intensity) as compared to beta-thalassemic red cells. The different abnormalities between alpha- and beta-thalassemic red cells may lead to different mechanism of red cell destruction and different severity of the disease.

Erythrocyte Deformability↗
PubMed1993

Cytometric analysis of blood cells from malaria-infected patients and in vitro infected blood.

Blood samples from malaria-infected patients and from in vitro culture were analyzed using the H*1 hematology analyzer. An attempt to find a hematologic parameter for detecting the malaria infection and to characterize the pathophysiological changes of red cells was made. The study included 18 malaria-infected patients (10 with Plasmodium falciparum and 8 with Plasmodium vivax) and 52 normal, healthy volunteers. Increased young large lymphocyte or large unstained cell count (LUC over 3%) in the peripheral blood of malaria-infected patients was evidence for malaria infection. Increased population dispersions of red cell volume (red cell distribution width: RDW) and intraerythrocytic hemoglobin concentration (hemoglobin distribution width: HDW) were demonstrated both in clinical samples and cultured blood. The increased RDW correlated with an increased percentage of macrocytes (r = 0.64, P = 0.004). Comparison of HDW and percentage of hypochromic red cells between the clinical specimen and the cultured blood supports the finding that changes in red cell hemoglobin concentration were mainly due to the response of the patient to malaria infection and partly due to the effect of malaria parasites on the red cells.

Cells, Cultured↗
PubMed1993

Dengue virus and endothelial cell: a related phenomenon to thrombocytopenia and granulocytopenia in dengue hemorrhagic fever.

Adhesion to endothelial cells by blood cells was assessed by measuring the cell number of each blood cell component in the supernatant after exposing blood cells to dengue-infected endothelial cells for 0, 10, 20 and 30 minutes. White blood cells, neutrophils, lymphocytes, platelets, and large lymphocytes or large unstained cells (LUC) preferentially bound to dengue-infected endothelial cells as compared to the control endothelial cells. P values were 0.0096 for total leukocytes and platelets, 0.006 for lymphocytes, and 0.001 for neutrophils and LUC. Monocytes basophils and eosinophils had no interaction with dengue-infected endothelial cells. The increased binding of neutrophil and platelet to endothelial cell may explain neutropenia and thrombocytopenia in DHF patients.

Agranulocytosis↗
PubMed1992

Reduced deformability of thalassemic erythrocytes and erythrocytes with abnormal hemoglobins and relation with susceptibility to Plasmodium falciparum invasion.

A number of genetically variant erythrocytes showed decreased deformability of both intact cells and membranes prepared therefrom as measured by laser diffractometry. Erythrocytes associated with minor or no clinical symptoms (eg, alpha-thalassemia traits, hemoglobin [Hb] E trait, Hb Constant Spring trait), which showed only a minimal decrease in deformability, were, in general, invaded efficiently by the malarial parasite Plasmodium falciparum. Other variant erythrocytes (beta-thalassemia/Hb E, homozygous Hb E, homozygous Hb Constant Spring, Hb H, Hb H/Hb Constant Spring) with low deformability showed different degrees of reduction in invasion susceptibility, most of which were less than proportional with deformability decrease. It is concluded that parasite invasion is only weakly related to gross cell deformability, which in turn depends on various factors other than membrane deformability.

Erythrocyte Deformability↗
PubMed1992

Erythrocyte volume and haemoglobin concentration in haemoglobin H disease: discrimination between the two genotypes.

Erythrocyte volume and haemoglobin concentration in individual red cells from 62 patients with Hb H disease: 37 with H genotype (alpha-thalassaemia 1/alpha-thalassaemia 2) and 25 H/CS genotype (alpha-thalassaemia 1/Hb Constant Spring) were measured using the H*1 haematology analyser. All 25 cases with H/CS genotype, the more severe genotype, had microcytes (red cells with a volume smaller than 60 fl) less than 35% and hypochromic red cells (red cells with haemoglobin concentration less than 28 g/dl) more than 35%. A discriminant function, the ratio between the percentage of hypochromic red cells and the percentage of microcytes (Hypo/Micro), was proposed. Most of the H/CS patients (76%) had Hypo/Micro greater than 2.5 whereas those of H patients (82%) were below 2.5. Red cell volume histograms were also characteristically different between the two genotypes: the H/CS had a peak between 60 and 90 fl while the H genotype showed a peak at or very close to 60 fl, indicating a greater degree of microcytosis. Increased hypochromia with a slight decrease in cell size of H/CS red cells suggests that the poor degree of haemoglobinization has no linkage or very little role in disturbing the synthesis of membrane proteins and their assembly to the plasma membrane.

Erythrocyte Indices↗
PubMed1992

Red blood cell and platelet induced differentiation of endothelial cells into capillary-like structure.

We cultivated endothelial cells of human umbilical vein origin in the presence of red blood cells and platelet-rich plasma, and observed the phenomena which occurred in the petri dish under phase contrast microscopy. Many small particles were observed after an overnight incubation. We washed the dish two times, then added thrombin to the dish. The network of thread-like strands appeared within 10 to 20 minutes of the addition, and at the same time the small particles adhered on the surface of the strands, swelled and fused gradually to cover the surface of the strands completely. Within 30 to 60 minutes the network of the strands changed into a capillary-like structure. These phenomena were not observed if we omitted red blood cells or platelet-rich plasma. Studies by transmission electron microscopy revealed that the inner surface of the lumen of the structure was covered with cells. The cells isolated from the lumen by trypsin grew to confluence in the conventional culture medium, and showed vWF antigen on their surface. These observations indicated that the method described is useful for in vitro study of angiogenesis.

Blood Platelets↗