Model Systems Employing Affinity Chromatography for Extraction of Toxic Substances Directly from Whole Blood

1993 ◽  
pp. 433-450 ◽  
Author(s):  
Shlomo Margel ◽  
Leon Marcus
Keyword(s):  
Affinity Chromatography ◽  
Whole Blood ◽  
Model Systems ◽  
Toxic Substances

Glycosylated hemoglobin measured by affinity chromatography: micro-sample collection and room-temperature storage.

1983 ◽  
Vol 29 (6) ◽  
pp. 1080-1082 ◽  
Author(s):  
R R Little ◽  
J D England ◽  
H M Wiedmeyer ◽  
D E Goldstein
Keyword(s):  
Affinity Chromatography ◽  
Whole Blood ◽  
High Performance ◽  
Room Temperature ◽  
Hemoglobin A1c ◽  
Glycosylated Hemoglobin ◽  
Exchange Chromatography ◽  
Sample Collection ◽  
Room Temperature Storage ◽  
Temperature Storage

Abstract Under proper conditions, whole blood can be stored at room temperature for as long as 21 days before measurement of glycosylated hemoglobin by affinity chromatography. Whole blood (anticoagulated with EDTA or heparin) was placed in capillary tubes, which were then sealed at both ends and stored at room temperature. Just before assay, whole blood was rinsed from the tubes and diluted 10-fold with water. Samples of each patient's blood were assayed as whole-blood hemolysates by affinity chromatography after zero, seven, 14, and 21 days of storage. Values for glycosylated hemoglobin did not change over 21 days of storage and values for each storage day correlated well (r = 0.97, p less than .0001) with hemoglobin A1C measured in fresh erythrocyte hemolysates by "high-performance" liquid ion-exchange chromatography.

scholarly journals Studying the processes of free-radical oxidation in patients with purulent-inflammatory diseases of soft tissues

2015 ◽  
Vol 96 (3) ◽  
pp. 302-306
Author(s):  
T Z Zakiev ◽  
S R Tuysin ◽  
O V Galimov ◽  
A R Gil’fanov ◽  
R D Sagdeev
Keyword(s):  
Free Radical ◽  
Whole Blood ◽  
Soft Tissues ◽  
Local Treatment ◽  
Free Radical Oxidation ◽  
Model Systems ◽  
Radical Oxidation ◽  
Tissue Homogenates

Aim. To study the effects of combined dressings on the processes of free radical oxidation in patients with purulent wounds.Methods. The performance of luminol-dependent chemiluminescence of whole blood from 30 healthy donors and 122 patients with purulent wounds, as well as tissue chemiluminescence of festering wounds tissue homogenates were examined before and after the treatment. To assess the free radical oxidation in vitro, spontaneous and zymosan-induced whole blood chemiluminescence measurement, iron-induced chemiluminescence assessment of festering wounds tissue homogenates were performed. The distribution of chemiluminescence parameters compared to normal expected distribution was analyzed to group homogeneity on these criteria.Results. Group of donors was homogeneous in composition that allowed the calculation of mean values. By changing in whole blood chemiluminescence of patients with soft tissue festering wounds, they were allocated to two groups, in which a marked increase or decline of the investigated parameters was seen. Increased luminol-dependent blood chemiluminescence indicates excessive free radicals generation by phagocytes and is characteristic of acute inflammation. Decreased intensity of chemiluminescence was observed in the blood of patients who showed reduced functional activity of phagocytes, which, together with the clinical features indicate a long-term smoldering inflammation. The comparative assessment of «Poliderm», «Voskopran», «Polysorb», «Levomekol» medications, used for local treatment of suppurative wounds, influence on free radical oxidation in model systems in vitro, was performed.Conclusion. There is a triple increase in chemiluminescence intensity in the acute phase of the inflammatory process, marking increased production of reactive oxygen radicals with microbicide activity); in long-term smoldering processes, chemiluminescence decreases by half, indicating reduced efficiency of protective mechanisms

Effect of Increased Tissue Factor Pathway Inhibitor (TFPI) Levels On Factor Xa Inhibition and Global Hemostasis in the Presence of TFPI-Antagonistic Aptamer BAX 499.

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 2207-2207 ◽  
Author(s):  
Michael Dockal ◽  
Rudolf Hartmann ◽  
Sabine Knappe ◽  
Michael Palige ◽  
Willibald Kammlander ◽  
...  
Keyword(s):  
Inhibitory Activity ◽  
Whole Blood ◽  
Plasma Levels ◽  
Thrombin Generation ◽  
Anticoagulant Activity ◽  
Reaction System ◽  
Factor Xa ◽  
Model Systems ◽  
Bleeding Tendency ◽  
Fold Excess

Abstract Abstract 2207 In a recent hemophilia clinical trial, patients' TFPI plasma levels substantially increased after intravenous or subcutaneous administration of TFPI-inhibitory aptamer BAX 499. To better understand the consequences of elevated plasma full-length (fl)-TFPI and the potency of BAX 499, the TFPI-inhibitory activity of a wide concentration range of BAX 499 was tested at levels up to 10 nM fl-TFPI in two model systems and in two global hemostatic coagulation assays. Factor Xa (FXa) inhibition assay measured the influence of BAX 499 on the interaction between TFPI and FXa. BAX 499 at 1000 nM efficiently inhibited fl-TFPI within the physiological concentration (< 0.5 nM). However, with increasing TFPI, inhibition of TFPI by BAX499 became considerably less efficient, reflected by increasing EC50 and decreasing maximum inhibition. At substantially elevated TFPI concentrations (10 nM) EC50 increased ∼20-fold, with TFPI retaining ∼70% of its FXa inhibitory activity. Similar results were observed in a more complex reaction system (TF-FVIIa-catalyzed FX activation). These results suggest that BAX 499 is a partial inhibitor of TFPI, efficiently neutralizing TFPI at low concentrations and less efficiently inhibiting TFPI at high TFPI levels. In thrombin generation assays with FVIII-inhibited plasma, a substantial excess of BAX 499 was required to compensate the anticoagulant activity of increasing fl-TFPI. Based on peak thrombin values, fl-TFPI levels that could be neutralized by a given BAX 499 concentration were determined. For example, a ∼50-fold excess of BAX 499 neutralized 0.2 nM TFPI added to FVIII-inhibited plasma, whereas a 140-fold excess was required for neutralization of 7.3 nM fl-TFPI added. An equivalent ROTEM experiment performed in FVIII-inhibited whole blood confirmed these findings. In summary, the data showed that even a high concentration of BAX 499 (1 μM) was not able to counteract the strong inhibitory effect of 10 nM fl-TFPI on FXa, on TF-FVIIa-catalyzed FX activation, on thrombin generation in plasma and clot formation in whole blood, due to the partially inhibitory properties of BAX 499. The current study offers an explanation for increased bleeding tendency in hemophilia patients associated with the ≥25-fold elevated fl-TFPI plasma levels after BAX 499 administration. BAX 499 does not appear to be able to compensate the anticoagulant activity of increased TFPI levels caused by partial inhibition in patients treated with BAX 499. Disclosures: No relevant conflicts of interest to declare.

Effects of whole blood storage on results for glycosylated hemoglobin as measured by ion-exchange chromatography, affinity chromatography, and colorimetry.

1983 ◽  
Vol 29 (6) ◽  
pp. 1113-1115 ◽  
Author(s):  
R R Little ◽  
J D England ◽  
H M Wiedmeyer ◽  
D E Goldstein
Keyword(s):  
Ion Exchange ◽  
Affinity Chromatography ◽  
Whole Blood ◽  
High Performance ◽  
Glycosylated Hemoglobin ◽  
Thiobarbituric Acid ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Sample Collection ◽  
Sample Stability

Abstract After storage of whole blood at either 4 or 20 degrees C, results for glycosylated hemoglobin by ion-exchange chromatography ("high-performance" liquid and mini-column chromatography), thiobarbituric acid colorimetry, and affinity chromatography were compared. At 4 degrees C, all methods gave acceptable results for samples stored for as long as a week. At 20 degrees C, the colorimetric and affinity methods also showed sample stability for a week or more. The ion-exchange methods were associated with a marked increase in values for glycosylated hemoglobin after a few days of storage. Evidently, care in details of sample collection and handling is especially important for ion-exchange methods, and the colorimetric and affinity methods have advantages over ion exchange in situations where long delays between sample collection and assay are unavoidable.

Characterization of toxicological effects of complex nano‐sized metal particles using in vitro human cell and whole blood model systems

2021 ◽  
Author(s):  
Linda Elfsmark ◽  
Barbro Ekstrand‐Hammarström ◽  
Nina Forsgren ◽  
Christian Lejon ◽  
Lars Hägglund ◽  
...  
Keyword(s):  
Human Cell ◽  
Whole Blood ◽  
Model Systems ◽  
Metal Particles ◽  
Toxicological Effects

scholarly journals Lipophilic Cations Rescue the Growth of Yeast under the Conditions of Glycolysis Overflow

Biomolecules ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1345
Author(s):  
Svyatoslav S. Sokolov ◽  
Ekaterina A. Smirnova ◽  
Olga V. Markova ◽  
Natalya A. Kireeva ◽  
Roman S. Kirsanov ◽  
...  
Keyword(s):  
Animal Model ◽  
Carbon Source ◽  
Oxidative Phosphorylation ◽  
Caloric Restriction ◽  
Inorganic Phosphate ◽  
Atp Synthesis ◽  
Model Systems ◽  
Rapid Screening ◽  
Ammonium Bromide ◽  
Toxic Substances

Chemicals inducing a mild decrease in the ATP/ADP ratio are considered as caloric restriction mimetics as well as treatments against obesity. Screening for such chemicals in animal model systems requires a lot of time and labor. Here, we present a system for the rapid screening of non-toxic substances causing such a de-energization of cells. We looked for chemicals allowing the growth of yeast lacking trehalose phosphate synthase on a non-fermentable carbon source in the presence of glucose. Under such conditions, the cells cannot grow because the cellular phosphate is mostly being used to phosphorylate the sugars in upper glycolysis, while the biosynthesis of bisphosphoglycerate is blocked. We reasoned that by decreasing the ATP/ADP ratio, one might prevent the phosphorylation of the sugars and also boost bisphosphoglycerate synthesis by providing the substrate, i.e., inorganic phosphate. We confirmed that a complete inhibition of oxidative phosphorylation alleviates the block. As our system includes a non-fermentable carbon source, only the chemicals that did not cause a complete block of mitochondrial ATP synthesis allowed the initial depletion of glucose followed by respiratory growth. Using this system, we found two novel compounds, dodecylmethyl diphenylamine (FS1) and diethyl (tetradecyl) phenyl ammonium bromide (Kor105), which possess a mild membrane-depolarizing activity.

Fine Structure of Platelet Interaction with a New Microcrystalline Collagen Preparation

1974 ◽  
Vol 32 ◽  
pp. 58-59
Author(s):  
W. H. Zucker ◽  
R. G. Mason
Keyword(s):  
Fine Structure ◽  
Platelet Aggregation ◽  
Hydrochloric Acid ◽  
Whole Blood ◽  
Platelet Adhesion ◽  
Hemostatic Agent ◽  
Native Collagen ◽  
Fibrillar Morphology ◽  
Clinical Interest ◽  
New Form

Platelet adhesion initiates platelet aggregation and is an important component of the hemostatic process. Since the development of a new form of collagen as a topical hemostatic agent is of both basic and clinical interest, an ultrastructural and hematologic study of the interaction of platelets with the microcrystalline collagen preparation was undertaken.In this study, whole blood anticoagulated with EDTA was used in order to inhibit aggregation and permit study of platelet adhesion to collagen as an isolated event. The microcrystalline collagen was prepared from bovine dermal corium; milling was with sharp blades. The preparation consists of partial hydrochloric acid amine collagen salts and retains much of the fibrillar morphology of native collagen.

3-D organization of fibrinogen receptors using computer reconstruction and whole-mount microscopy

1988 ◽  
Vol 46 ◽  
pp. 30-31
Author(s):  
E. T. O'Toole ◽  
R. R. Hantgan ◽  
J. C. Lewis
Keyword(s):  
Whole Blood ◽  
Colloidal Gold ◽  
Blood Platelets ◽  
Cell Contact ◽  
Computer Assisted ◽  
Adherent Cells ◽  
Differential Centrifugation ◽  
Computer Reconstruction ◽  
Sds Page ◽  
Whole Mount

Thrombocytes (TC), the avian equivalent of blood platelets, support hemostasis by aggregating at sites of injury. Studies in our lab suggested that fibrinogen (fib) is a requisite cofactor for TC aggregation but operates by an undefined mechanism. To study the interaction of fib with TC and to identify fib receptors on cells, fib was purified from pigeon plasma, conjugated to colloidal gold and used both to facilitate aggregation and as a receptor probe. Described is the application of computer assisted reconstruction and stereo whole mount microscopy to visualize the 3-D organization of fib receptors at sites of cell contact in TC aggregates and on adherent cells.Pigeon TC were obtained from citrated whole blood by differential centrifugation, washed with Ca++ free Hank's balanced salts containing 0.3% EDTA (pH 6.5) and resuspended in Ca++ free Hank's. Pigeon fib was isolated by precipitation with PEG-1000 and the purity assessed by SDS-PAGE. Fib was conjugated to 25nm colloidal gold by vortexing and the conjugates used as the ligand to identify fib receptors.

Nuclear coiled bodies and the nucleolus

1994 ◽  
Vol 52 ◽  
pp. 20-21
Author(s):  
K. Brasch ◽  
J. Williams ◽  
D. Gallo ◽  
T. Lee ◽  
R. L. Ochs
Keyword(s):  
Mouse Liver ◽  
Nuclear Body ◽  
Nuclear Bodies ◽  
Model Systems ◽  
Coiled Body ◽  
Rrna Processing ◽  
Malignant Cells ◽  
Sm Proteins ◽  
Coiled Bodies ◽  
Unique Protein

Though first described in 1903 by Ramon-y-Cajal as silver-staining “accessory bodies” to nucleoli, nuclear bodies were subsequently rediscovered by electron microscopy about 30 years ago. Nuclear bodies are ubiquitous, but seem most abundant in hyperactive and malignant cells. The best studied type of nuclear body is the coiled body (CB), so termed due to characteristic morphology and content of a unique protein, p80-coilin (Fig.1). While no specific functions have as yet been assigned to CBs, they contain spliceosome snRNAs and proteins, and also the nucleolar protein fibrillarin. In addition, there is mounting evidence that CBs arise from or are generated near the nucleolus and then migrate into the nucleoplasm. This suggests that as yet undefined links may exist, between nucleolar pre-rRNA processing events and the spliceosome-associated Sm proteins in CBs.We are examining CB and nucleolar changes in three diverse model systems: (1) estrogen stimulated chick liver, (2) normal and neoplastic cells, and (3) polyploid mouse liver.

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