Biomedical subjects

A L Inyang

Publications and source records attributed to A L Inyang.

10 recordsLinked to original sources

PubMed1995

Decrease in platelet survival and total platelet sialic acid concentration in rats infected with Plasmodium bergei bergei.

Suckling Wistar rats aged 3-5 weeks were infected through their dorsal tail vein with P. berghei berghei passed in Swiss albino mice. Platelet recovery and platelet survival using 51Cr-labelled heterologous platelets obtained from adult Wistar rats were determined in the infected animals on different post-infection days and on a group of non-infected rats as controls. Total platelet sialic acid was also determined in the same groups of animals. The results showed reduced platelet recovery, shortened survival and reduced total platelet sialic acid content in the infected animals compared with control values. The reduction in total platelet sialic acid content was related to the degree of parasitaemia and reached significant levels on the 5th post-infection day. It is concluded that the shortened platelet survival and reduced total platelet sialic acid content observed in the P. berghei infected rats were causally related and may account for the thrombocytopaenia reported in experimental and natural malaria infections of animals and man.

Animals↗

PubMed1990

Tissue-plasminogen activator stimulates endothelial cell migration in wound assays.

The ability of tissue plasminogen activator (tPA) to induce human umbilical vein endothelial (HUVE) cell migration was studied using an in vitro, serum-free wound assay system. At pharmacological doses, tPA stimulated HUVE cell migration dose-dependently. Treatment of cells with epsilon amino caproic acid (EACA) to detach cell-surface and extracellular matrix bound plasminogen, which could lead to plasmin generation, resulted in increased HUVEcell migration on stimulation with tPA.Plasminogen activator inhibitor-1 (PAI-1), a natural plasminogen activator inhibitor, abolished tPA-induced HUVEcell migration. These results demonstrate for the first time that tPA is capable of stimulating endothelial cell migration in wound assays and this effect is susceptible to PAI-1 inhibition.

Cell Movement↗

PubMed1990

Basic fibroblast growth factor expression in human omental microvascular endothelial cells and the effect of phorbol ester.

The human omentum contains a potent, not yet identified angiogenic activity. The omentum is very vascularized. Therefore, we investigated whether human omental microvascular endothelial cells (HOME cells) express the angiogenic peptide basic fibroblast growth factor (bFGF). Cytosol prepared from HOME cells stimulated DNA synthesis in bovine epithelial lens cells (BEL cells). The mitogenic activity could be neutralized by an anti-bFGF antibody. Basic FGF-like material from the HOME cell cytosol was bound onto a heparin-Sepharose column at 0.6 M and was eluted at 3 M NaCl. The 3 M NaCl eluted material reacted with the specific anti-bFGF antibody in an ELISA and stimulated DNA synthesis. It did not react with a specific anti-acidic fibroblast growth factor (aFGF) antibody. Western blotting experiments using the same bFGF antibody showed the presence of a major band of 17 Kd and a doublet of 20-22 Kd. Northern blotting of non-stimulated HOME cells using a specific 1.4 kb bFGF probe showed the presence of 5 molecular species of 6.6, 3.7, 2.2, 2.0, and 1.0 kb. No aFGF mRNA was detected with a specific previously characterized 4.04 kb probe. 12-O-tetradecanoylphorbol 13-acetate (TPA) did not influence significantly the expression of bFGF at the protein and mRNA level in HOME cells. Thus, protein kinase C activation by TPA did not appear to modulate significantly the expression of bFGF for that cell type. Contrastingly, human umbilical vein endothelial cells (HUVE cells), which expressed no bFGF and aFGF mRNA at a basal level, were induced to express bFGF but not aFGF mRNA when stimulated by TPA. These results suggest that the described angiogenic activity could be the bFGF-like mitogen contained in HOME cells and that these cells are different from endothelial cells derived from large vessels (HUVE cells) regarding the expression of bFGF.

Biological Assay↗

PubMed1990

Chloroquine's modulation of endothelial cell activity induced with basic fibroblast growth factor and human serum: effect on mitogenesis, protease production and cell migration.

Chloroquine modulates the activity of cultured human microvascular endothelial (HOME) cells in a complex fashion. At concentrations of 5-25 microM, CQ inhibits basic fibroblast growth factor (bFGF-) and human serum-induced mitogenic activity in these cells, in a dose-dependent manner. The kinetics of CQ's inhibitory actions on serum-induced mitogenesis in HOME cells slowly develops with only 30% of maximum inhibition reached after 24 hours. In HOME cells grown in serum-free medium, CQ raised tissue-plasminogen activator antigen levels in cell extracts. There was also a potentiation of bFGF-induced t-PA production. The kinetics of CQ's stimulatory effect on t-PA production by HOME cells, suggest that this effect precedes its inhibitory actions on mitogenesis. This effect of CQ on t-PA generation in endothelial cells was susceptible to cycloheximide inhibition. In wound assays, HOME cell migration, induced with bFGF and HS, was potentiated by CQ.

Blood Proteins↗

PubMed1990

Platelet aggregation inhibition in Trypanosoma vivax infection of sheep.

We have investigated in vitro platelet aggregation in platelet rich plasma from Trypanosoma vivax infected and control sheep using the dual channel Payton Aggregometer. Final concentrations of the following inducing agents were used: 1.2 um ADP, 6.2 ug collagen, 1.2 ug ristocetin and 1 u thrombin. These showed that there was a significantly reduced aggregation of platelets from infected sheep (13.4 +/- 1.1 pc at week 3 post infection when compared with control sheep PRP 95.0 +/- 1.0pc; P less than 0.001) using ADP. Similar differences were also obtained with other inducing agents. Preliminary 14C-5HT uptake and release studies showed that there was difference in the uptake of label between platelets from infected (18.6pc) and control (28.4pc) sheep. However, when release was inducted, comparable results were obtained for both infected and control sheep platelets. It is concluded that the degree of aggregation inhibiting varies directly with the level of parasitaemia.

Animals↗

PubMed1989

Binding, internalization, and degradation of basic fibroblast growth factor in human microvascular endothelial cells.

The binding, internalization, and degradation of basic fibroblast growth factor (bFGF) in human omental microvascular endothelial cells (HOME cells) were investigated. Binding studies of bFGF in human endothelial cells have not yet been reported. Basic FGF bound to HOME cells (KD of 42.0 +/- 3.8 pM and 70,526 +/- 6121 binding sites/cell for the high-affinity sites, KD of 0.933 +/- 0.27 nM and 630,252 +/- 172,459 sites/cell for low-affinity binding sites). The number of low-affinity binding sites was found to be variable. Washing the cells with 2 M phosphate-buffered saline removed completely 125I-bFGF bound to low-affinity binding sites but decreased also the high-affinity binding. The majority of the surface-bound 125I-bFGF was removed by washing the cells with acetic acid buffer at pH 3. At 37 degrees C, 30% of the cell-associated 125I-bFGF became resistant to the acidic wash after 90 min, suggesting that this fraction of bound 125I-bFGF was internalized. At this temperature, degradation of the internalized ligand was followed after 1 h by the appearance of three major bands of 15,000, 10,000, and 8,000 Da and was inhibited by chloroquine. These results demonstrated two classes of binding sites for bFGF in HOME cells; the number of high-affinity binding sites being larger than the number reported for bovine capillary endothelial cells. The intracellular processing of bFGF in HOME cells seems to be different from that of heparin binding growth factor-1 in murine lung capillary endothelial cells and of eye-derived growth factor-1 in Chinese hamster fibroblasts.

Binding Sites↗

PubMed1989

Responses of the isolated rectum of the rainbow lizard (Agama agama) to sympathomimetics.

The relaxant effects of isoprenaline, noradrenaline, and adrenaline on the isolated rectum of the rainbow lizard (Agama agama) were studied. Responses were measured as a reduction of carbachol-induced contractions for each sympathomimetic agent. Isoprenaline, adrenaline, noradrenaline produced a dose-dependent relaxation of this preparation and the order of potency was as given. The pD2 value of 8.15 +/- 1.88 obtained for isoprenaline was significantly different (p less than 0.05) from those for adrenaline (5.80 +/- 0.90) and noradrenaline (5.25 +/- 1.18). H35/25, propranolol, and practolol competitively antagonized the relaxant effects of isoprenaline on the isolated lizard rectum. The pA2 values for these beta-adrenoceptor antagonists did not differ significantly (at p less than 0.05). alpha-Adrenoceptor antagonists, phentolamine and phenoxybenzamine, failed to alter the relaxant responses of these sympathometics to any appreciable extent. These results are interpreted to suggest that the relaxant effect produced by these sympathomimetics are mediated predominantly by beta-adrenoceptors that are not significantly differentiated into subtypes. alpha-Adrenoceptors in this preparation contribute minimally to the observed inhibitory response following sympathomimetic stimulation.

Adrenergic alpha-Antagonists↗

PubMed1989

Platelet-aggregating activity of released factor(s) from Trypanosoma brucei brucei.

The effect of factors derived from Trypanosoma brucei brucei on rat platelets was studied. T. brucei at a concentration of 4 X 10(9) trypanosomes/ml phosphate saline glucose (PSG) was stored at -20 degrees C for 18 h, thawed, and a supernatant fraction, trypanosome-derived supernatant (TDS) was obtained by spinning the sample at 3000 g for 10 min at 20 degrees C. Normal rat platelets, prepared as platelet-rich plasma (PRP), were then incubated with TDS in the absence or presence of ADP (0.05-0.1 microM). The results showed that approximately 83% platelet aggregation was induced by addition of TDS (50 microliters; 113 micrograms protein) to 100 microliters PRP with a platelet count of 10(6). simultaneous addition of ADP and TDS to PRP produced a synergistic effect. It was also shown that a supernatant fraction, obtained by incubating live T. brucei (4 X 10(9)/microliters PSG) at 0 degrees C 1 h and spinning down the trypanosomes (3000 g for 10 min), also induced platelet aggregation. The nature of the factor(s) derived from, or released by, T. brucei inducing platelet aggregation is being investigated but it has been shown not to be ADP.

Animals↗

PubMed1987

Platelet reactions after interaction with cultured Plasmodium falciparum infected erythrocytes.

An in vitro model for studying the interaction between normal human platelets and Plasmodium falciparum infected erythrocytes in culture is described. After the interaction, changes in platelet function such as enhanced aggregation response to exogenous ADP and increased secretion of dense granule contents were reproduced. Some of these responses represented manifestations of platelet hypersensitivity described earlier in acute malaria infections in man and mice. Preliminary investigations of the mechanisms involved in such reactions revealed that ADP and thromboxane A2 mechanisms contributed about 79% and 18.5% of the enhanced aggregation response to exogenous stimuli in the system.

Adenosine Diphosphate↗

PubMed1987

Platelet reactions in acute Plasmodium berghei infection in Swiss albino mice.

Swiss albino mice were infected by the intraperitoneal route with P. berghei berghei malaria parasite, and platelets, white cell counts and some coagulation parameters were monitored in order to find out whether changes reported in man also occurred in the mice. Parasitaemia developed form the 2nd post-infection day and reached significant levels by the 4th-6th day. Reduced circulating platelets which reached severe thrombocytopenic levels were observed. parallel with the increasing degree of parasitaemia. Anaemia which progressed to severe degree was also observed as was a slight leucocytosis attributed to the presence of normal mouse erythrocytes in the peritoneal space. All untreated animals died by the 6th day of infection. Intramuscular chloroquine sulphate (20 micrograms/g body wt.) given for 7 days completely cured the malaria, and white cell and platelet counts were restored to preinfection levels in each animal about 2 weeks after treatment had ceased. Platelet hypersensitivity to exogenous ADP was observed within 48 hours of infection and persisted with the parasitaemia. Prothrombin time (PT) and activated partial thromboplastin time (APTT) were prolonged while clottable fibrinogen concentration was reduced.

Animals↗