REACTION OF 14C-ACETALDEHYDE WITH WHOLE BLOOD IN VITRO: FURTHER EVIDENCE FOR THE FORMATION OF UNSTABLE COMPLEXES WITH PLASMA PROTEINS AND RED CELLS

1994 ◽  
Keyword(s):  
Whole Blood ◽  
Plasma Proteins ◽  
Red Cells

scholarly journals The effect of the plasticizer di-2-ethylhexyl phthalate on the survival of stored RBCs

Blood ◽  
1988 ◽  
Vol 71 (2) ◽  
pp. 448-452 ◽  
Author(s):  
JP AuBuchon ◽  
TN Estep ◽  
RJ Davey
Keyword(s):  
Whole Blood ◽  
Red Cells ◽  
Refrigerated Storage ◽  
Plasticized Pvc ◽  
Normal Human ◽  
Plastic Containers ◽  
Ethylhexyl Phthalate ◽  
Citrate Phosphate

Abstract Recent in vitro studies have shown that di-2-ethylhexyl-phthalate (DEHP) inhibits the deterioration of RBCs during refrigerated storage in containers that use this compound as a plasticizer. The experiments described in this report were designed to assess whether this in vitro protective effect of DEHP would result in a prolonged in vivo survival of RBCs infused into normal human recipients. Whole blood collected from ten normal donors was stored for 35 days in citrate-phosphate- dextrose-adenine (CPDA-1) anticoagulant contained in polyvinylchloride (PVC) bags plasticized with DEHP or a trimellitate compound that is known to have low leachability. Aliquots of RBCs from each container were then labeled with chromium-51 and were reinfused into the original donors. For blood stored in DEHP-plasticized PVC bags, 24% more red cells survived in vivo 24 hours after reinfusion than was observed when the blood had been stored in trimellitate-plasticized bags (P less than .001). Whole blood stored in glass bottles showed a similar improvement in in vivo survival when DEHP was added in weekly increments to mimic the accumulation of this plasticizer seen during storage in plastic containers. Survival of packed red cells stored in the presence of DEHP increased by 14% compared with storage in trimellitate-plasticized bags (P less than .05). In agreement with previous studies, hemolysis and microvesicle formation were also reduced in the presence of DEHP. These results suggest that proposed new storage systems lacking DEHP should be carefully evaluated to determine whether adequate post-transfusion survival of RBCs may be achieved.

Reduction of Prion Infectivity in Packed Red Cells by Separation of Whole Blood and Washing of the Red Cell Fraction.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 2890-2890
Author(s):  
Rodrigo Morales ◽  
Kimberley A. Buytaert-Hoefen ◽  
Eric T. Hansen ◽  
Dennis Hlavinka ◽  
Raymond Goodrich ◽  
...  
Keyword(s):  
Blood Cell ◽  
Red Blood Cell ◽  
Whole Blood ◽  
Red Cells ◽  
Cell Fraction ◽  
Health Concern ◽  
Red Cell ◽  
New Variant

Abstract Although prion diseases are rare in humans, the established link between a new variant form of CJD (vCJD) and the consumption of cattle meat contaminated by BSE have raised concerns about a possible outbreak of a large epidemic in the human population. Over the past few years, BSE has become a significant health concern in several countries, and it now seems apparent that vCJD can also be iatrogenically transmitted from human to human by blood transfusion. Exacerbating this state of affairs is the lack of a reliable test to identify individuals incubating the disease during the long and silent period from the onset of infection to the appearance of clinical symptoms. The purpose of this research study was to evaluate the effectiveness of separation of whole blood and washing of the red cell fraction for the removal of infectious scrapie prion protein (PrPSc) from red blood cell (RBC) suspensions. Samples of human, whole blood were spiked with 5 × 106 LD50 263K PrPSc. Analysis of the treated sample supernatants by Western blot revealed that approximately >88% of the PrPSc was removed with the initial plasma expression and the equivalent of 6% was detected in a saline wash (300 mL; 0.9% saline). The final sample of RBCs revealed no detectable levels of PrPSc by Western blots. Further analysis of the treated RBCs using the PMCA assay indicated detectable amounts of PrPSc only after 2 consecutive amplification rounds. Semi-quantitative analysis of PMCA amplification enabled us to estimate that the treated RBCs contained less than 1 × 104 LD50 PrPSc. This corresponded to removal levels exceeding ≥99% of spiked material in whole blood. These in vitro estimations were confirmed by in vivo infectivity studies in a hamster model of disease transmission. Results from in vivo studies displayed significant differences in the incubation periods of the spiked blood inoculated hamsters (100.1 ± 1.7) versus washed RBCs (135.8 ± 6.7). Moreover, a substantial difference in the attack rate (6/15: 40% in washed RBC, versus 13/13: 100% in spiked blood) further indicated a substantial removal of infectious prions. Comparison of this data with results of animals inoculated with different dilutions of infectious material, indicated a >99.94% reduction of infectivity. Washed, packed human red cells produced by this procedure were able to be stored in standard additive solutions (AS-3) for 42 days while still meeting all in vitro blood bank standards for acceptable red cell quality. Conclusion This data suggests that separation of plasma coupled with a simple, low volume wash of red cells may represent an efficient method to remove prions from red blood cell fractions, thus reducing possible infectivity of these products.

EFFECT OF PLASMA PROTEINS ON THE FRAGILITY OF ERYTHROCYTES

1966 ◽  
Vol 44 (3) ◽  
pp. 401-407 ◽  
Author(s):  
D. C. Triantaphyllopoulos ◽  
E. H. Krikke
Keyword(s):  
Protective Effect ◽  
Plasma Proteins ◽  
Structural Integrity ◽  
Red Cells ◽  
The Other ◽  
Blood Protein ◽  
Mechanical Trauma ◽  
Fresh Blood ◽  
Overnight Incubation

Addition of fibrinogen or of the anticoagulant fraction of incubated fibrinogen (AFIF) to fresh blood in vitro decreases both the osmotic and the mechanical fragility of the erythrocytes. This suggests that these proteins contribute to the structural integrity of the red cells. On the other hand, addition of albumin does not affect significantly the fragility. Overnight incubation of the blood–protein mixtures abolishes the protective effect of fibrinogen on both kinds of fragility and that of AFIF on the osmotic type. The increased resistance of the red cells, however, to mechanical trauma in the presence of AFIF remains unaffected by incubation.

Kinetics of chlorambucil in vitro: effects of fluid matrix, human gastric juice, plasma proteins and red cells

1997 ◽  
Vol 103 (3) ◽  
pp. 187-198 ◽  
Author(s):  
K. Löf ◽  
J. Hovinen ◽  
P. Reinikainen ◽  
L.M. Vilpo ◽  
E. Seppälä ◽  
...  
Keyword(s):  
Gastric Juice ◽  
Plasma Proteins ◽  
Red Cells ◽  
Human Gastric Juice ◽  
In Vitro Effects ◽  
Kinetics Of

scholarly journals Development of a mass-spectrometry based method for the identification of the in vivo whole blood and plasma ADP-ribosylomes

2020 ◽  
Author(s):  
Stephanie C. Lüthi ◽  
Anna Howald ◽  
Kathrin Nowak ◽  
Robert Graage ◽  
Giody Bartolomei ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Whole Blood ◽  
Sus Scrofa ◽  
Plasma Proteins ◽  
Post Translational Modification ◽  
Blood Samples ◽  
Adp Ribosylation

ABSTRACTBlood and plasma proteins are heavily investigated as biomarkers for different diseases. However, the post-translational modification states of these proteins are rarely analyzed since blood contains many enzymes that rapidly remove these modification after sampling. In contrast to the well-described role of protein ADP-ribosylation in cells and organs, its role in blood remains mostly uncharacterized. Here, we discovered that plasma phosphodiesterases and/or ADP-ribosylhydrolases rapidly demodify in vitro ADP-ribosylated proteins. Thus, to identify the in vivo whole blood and plasma ADP-ribosylomes, we established a novel mass-spectrometry based workflow that was applied to blood samples collected from LPS-treated pigs (Sus scrofa), which serves as a model for human systemic inflammatory response syndrome. These analyses identified 60 ADP-ribosylated proteins, 17 of which were ADP-ribosylated plasma proteins. This new protocol provides an important step forward for the rapidly developing field of ADP-ribosylation and defines the blood and plasma ADP-ribosylomes under both healthy and disease conditions.

scholarly journals The effect of the plasticizer di-2-ethylhexyl phthalate on the survival of stored RBCs

Blood ◽  
1988 ◽  
Vol 71 (2) ◽  
pp. 448-452 ◽  
Author(s):  
JP AuBuchon ◽  
TN Estep ◽  
RJ Davey
Keyword(s):  
Whole Blood ◽  
Red Cells ◽  
Refrigerated Storage ◽  
Plasticized Pvc ◽  
Normal Human ◽  
Plastic Containers ◽  
Ethylhexyl Phthalate ◽  
Citrate Phosphate

Recent in vitro studies have shown that di-2-ethylhexyl-phthalate (DEHP) inhibits the deterioration of RBCs during refrigerated storage in containers that use this compound as a plasticizer. The experiments described in this report were designed to assess whether this in vitro protective effect of DEHP would result in a prolonged in vivo survival of RBCs infused into normal human recipients. Whole blood collected from ten normal donors was stored for 35 days in citrate-phosphate- dextrose-adenine (CPDA-1) anticoagulant contained in polyvinylchloride (PVC) bags plasticized with DEHP or a trimellitate compound that is known to have low leachability. Aliquots of RBCs from each container were then labeled with chromium-51 and were reinfused into the original donors. For blood stored in DEHP-plasticized PVC bags, 24% more red cells survived in vivo 24 hours after reinfusion than was observed when the blood had been stored in trimellitate-plasticized bags (P less than .001). Whole blood stored in glass bottles showed a similar improvement in in vivo survival when DEHP was added in weekly increments to mimic the accumulation of this plasticizer seen during storage in plastic containers. Survival of packed red cells stored in the presence of DEHP increased by 14% compared with storage in trimellitate-plasticized bags (P less than .05). In agreement with previous studies, hemolysis and microvesicle formation were also reduced in the presence of DEHP. These results suggest that proposed new storage systems lacking DEHP should be carefully evaluated to determine whether adequate post-transfusion survival of RBCs may be achieved.

scholarly journals The in Vitro Bleeding Time

1977 ◽  
Author(s):  
J.A. Blakely ◽  
M. F. X. Glynn ◽  
J. Prchal
Keyword(s):  
Whole Blood ◽  
Bleeding Time ◽  
Red Cells ◽  
Von Willebrand's Disease ◽  
Flow Rates ◽  
Shed Blood ◽  
Time Required ◽  
Teflon Tubing ◽  
Fibrin Thrombus

The observation by Didisheim of platelet-fibrin hemostatic plugs occluding puncture sites in teflon AV shunts prompted an attempt to produce similar plugs in vitro using a pump to circulate shed blood. Various anticoagulants, tubing types, flow rates, pressures and methods of producing suitable holes have been investigated. Satisfactory results have been obtained with drilled holes 0.013 - 0.018” diameter in tygon or teflon tubing using heparinized or native, but not EDTA or citrated whole blood. Holes are occluded by platelet fibrin thrombus. Red cells are necessary for plugs to form but are not a component of the plug. Time required for plugs to form correlate with the template bleeding time and are prolonged by thrombocytopenia, aspirin administration, and Von Willebrand’s disease. Possible applications are being explored.

Immunoenzymatic demonstration of the simultaneous presence of transferrin, hemopexin and albumin in a same hepatocyte and their ultrastructural localization

1978 ◽  
Vol 36 (2) ◽  
pp. 88-89
Author(s):  
M. Kraemer ◽  
J. Foucrier ◽  
J. Vassy ◽  
M.T. Chalumeau
Keyword(s):  
Plasma Proteins ◽  
Rat Hepatocytes ◽  
Ultrastructural Localization ◽  
Carrier Proteins ◽  
Normal Cells ◽  
Adult Rat ◽  
Adult Rat Hepatocytes ◽  
Monospecific Antisera ◽  
Different Levels

Some authors using immunofluorescent techniques had already suggested that some hepatocytes are able to synthetize several plasma proteins. In vitro studies on normal cells or on cells issued of murine hepatomas raise the same conclusion. These works could be indications of an hepatocyte functionnal non-specialization, meanwhile the authors never give direct topographic proofs suitable with this hypothesis.The use of immunoenzymatic techniques after obtention of monospecific antisera had seemed to us useful to bring forward a better knowledge of this problem. We have studied three carrier proteins (transferrin = Tf, hemopexin = Hx, albumin = Alb) operating at different levels in iron metabolism by demonstrating and localizing the adult rat hepatocytes involved in their synthesis.Immunological, histological and ultrastructural methods have been described in a previous work.

Investigation of the Free and Total Thyroxine-binding Capacity of Plasma Proteins in Thyroid Disorders

1971 ◽  
Vol 10 (04) ◽  
pp. 299-304
Author(s):  
József Takó ◽  
János Fischer ◽  
Jusztina Juhász ◽  
Ilona Sztraka ◽  
István Kapus ◽  
...  
Keyword(s):  
Blood Cell ◽  
Thyroid Function ◽  
Oral Contraceptives ◽  
Red Blood Cell ◽  
Plasma Proteins ◽  
Binding Capacity ◽  
Thyroid Disorders ◽  
Thyroid Function Tests ◽  
Total Thyroxine

SummaryThe results of thyroid function tests have been compared with data on the thyroxine-binding capacity of plasma proteins in hyper-, hypo- and euthyroid cases, the latter including women taking oral contraceptives (Infecundin). It was found that there exists a significant correlation of exponential nature between the in vitro red blood cell 125I-triiodothyronine uptake (RCU) and the free thyroxine-binding capacity of the thyroxine-inding globulin (TBG).

Effect of Dipyridamole on Spontaneous Platelet Aggregation in Whole Blood Decreases with the Time After Venepuncture: Evidence for the Role of ADP

1987 ◽  
Vol 58 (02) ◽  
pp. 744-748 ◽  
Author(s):  
A R Saniabadi ◽  
G D O Lowe ◽  
J C Barbenel ◽  
C D Forbes
Keyword(s):  
Platelet Aggregation ◽  
Correlation Coefficient ◽  
Whole Blood ◽  
Blood Platelet ◽  
Inhibitory Effect ◽  
Time Interval ◽  
Adp Receptor ◽  
Spontaneous Platelet Aggregation

SummarySpontaneous platelet aggregation (SPA) was studied in human whole blood at 3, 5, 10, 20, 30, 40 and 60 minutes after venepuncture. Using a whole blood platelet counter, SPA was quantified by measuring the fall in single platelet count upon rollermixing aliquots of citrated blood at 37° C. The extent of SPA increased with the time after venepuncture, with a correlation coefficient of 0.819. The inhibitory effect of dipyridamole (Dipy) on SPA was studied: (a) 10 μM at each time interval; (b) 0.5-100 μM at 3 and 30 minutes and (c) 15 μM in combination with 100 μM adenosine, 8 μM 2-chloroadenosine (2ClAd, an ADP receptor blocker) and 50 μM aspirin. There was a rapid decrease in the inhibitory effect of Dipy with the time after venepuncture; the correlation coefficient was -0.533. At all the concentrations studied, Dipy was more effective at 3 minutes than at 30 minutes after venepuncture. A combination of Dipy with adenosine, 2ClAd or aspirin was a more effective inhibitor of SPA than either drug alone. However, when 15 μM Dipy and 10 μM Ad were added together, the inhibitory effect of Dipy was not increased significantly, suggesting that Dipy inhibits platelet aggregation independent of Ad. The increase in SPA with the time after venepuncture was abolished when blood was taken directly into the anticoagulant containing 5 μM 2ClAd. It is suggested that ADP released from the red blood cells is responsible for the increased platelet aggregability with the time after venepuncture and makes a serious contribution to the artifacts of in vitro platelet function studies.

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