The Effect of Methylene Blue Addition to Whole Blood during Prolonged Storage

Vox Sanguinis ◽  
1972 ◽  
Vol 22 (3) ◽  
pp. 236-243
Author(s):  
W.F. Kocholaty ◽  
R.B. Dawson jr.
Keyword(s):  
Methylene Blue ◽  
Whole Blood ◽  
Prolonged Storage

The Effect of Methylene Blue Addition to Whole Blood during Prolonged Storage

Vox Sanguinis ◽  
1972 ◽  
Vol 22 (3) ◽  
pp. 236-243 ◽  
Author(s):  
W. F. Kocholaty ◽  
R. B. Dawson
Keyword(s):  
Methylene Blue ◽  
Whole Blood ◽  
Prolonged Storage

Forensic Electrochemical Presumptive Blood Test Based on the Voltammetric Behaviour of Methylene Blue and Whole Blood

2021 ◽  
Author(s):  
Sarah Cook ◽  
Kevin C. Honeychurch
Keyword(s):  
Methylene Blue ◽  
Whole Blood ◽  
Blood Test ◽  
Archaeological Sciences

The ability to identify the presence of blood residues is important in a number of fields, such as in the forensic and archaeological sciences. A number of tests presently exist;...

scholarly journals Effect of Storage on Levels of Nitric Oxide Derivatives in Blood Components

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 999-999
Author(s):  
Fabiola Grizzatti ◽  
Melissa A Qazi ◽  
David Stroncek ◽  
Nathawut Sibmooh ◽  
Barbora Piknova ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Whole Blood ◽  
Blood Product ◽  
Nitrate Concentration ◽  
Extracellular Fluid ◽  
Prolonged Storage ◽  
Blood Components ◽  
Transfusion Therapy ◽  
Xanthine Oxidase Inhibitor ◽  
Duration Of Storage

Abstract In light of recent papers stressing the importance of decreased levels of SNO-hemoglobin (SNO-Hb) to the pronounced nature of deleterious effects of transfusion of stored red blood cells (RBCs), there has been an increased interest in the practice of blood storage. Dejam et al. (Blood, 2005) previously demonstrated the critical nature of RBCs in NO physiology: they serve as the major intravascular stores of nitrite, which is eventually converted to NO, an important player in vasoregulation. The purpose of this study is to quantify the NO metabolites, nitrite and nitrate, in three blood components and evaluate their levels over time of storage. Blood obtained from 6 healthy volunteer donors was split into whole blood, leukoreduced, and non-leukoreduced packed RBCs and stored in polyvinyl chloride (PVC) bags for 42 days at 4°C. PVC bags were maintained in either room air or an argon chamber. Nitrite, nitrate, and SNO-Hb/nitrosyl-hemoglobin (HbNO) were measured using reductive gas-phase chemiluminescence. In all blood components, the nitrite and nitrate were detected in higher concentrations in RBCs than in the extracellular fluid compartment. Mean nitrite value immediately before storage was 152±13nM, but fell rapidly upon storage. Nitrite levels continued to decrease with storage time, while nitrate levels remained constant for the duration of storage. In the leukoreduced blood product, nitrite levels were 75±8nM on day 1 and 50±9nM by day 42; the concentration of nitrate in the leukoreduced blood product was 34±3uM on day 1 and 34±4uM on day 42. The nitrite levels in non-leukoreduced blood product were 76±12nM on day 1 and 37±7 by day 42; the nitrate concentration in the non-leukoreduced blood product was 35±3uM on day 1 and 32±0.4uM on day 42. In whole blood, nitrite levels were 64±11nM on day 1 and 44±9nM by day 42; the nitrate concentration was 47±2uM on day 1 and 43±6uM on day 42. SNO-Hb levels were very low in fresh blood and virtually undetectable after one day of storage. Interestingly, nitrite levels never reached zero. Enzyme inhibitors—L-NAME (nitric oxide synthase inhibitor), acetazolamide (carbonic anhydrase inhibitor), and oxypurinol (xanthine oxidase inhibitor)—did not lower nitrite levels enough to explain the remaining nitrite present in the PVC bags after 42 days. pH decreased slightly, while pO2 increased in all three components during storage; this is likely due to the diffusion of oxygen from room air into the PVC bags. Control experiments with saline showed an increase in nitrite levels, while nitrate levels remained stable over 42 days. When stored in an argon chamber, both blood and saline samples showed relatively lower nitrite levels than their room air counterparts. Thus, during blood bank storage, nitrite levels decrease in blood while nitrate levels remain stable. The diffusion of nitrogenous gases may explain why nitrite does not completely disappear under standard storage conditions. Our results suggest that most of the NO pathway is initially retained, but greater changes occur with prolonged storage. These measurements of NO derivatives may have implications for transfusion therapy, explaining some of the adverse effects seen with RBC transfusion and providing a foundation for enhancing blood preservation through improvement of storage practices.

STUDIES OF SERUM ELECTROLYTE CHANGES DURING EXCHANGE TRANSFUSION

PEDIATRICS ◽  
1954 ◽  
Vol 13 (5) ◽  
pp. 412-418
Author(s):  
GERALD MILLER ◽  
AUGUSTA B. MCCOORD ◽  
HOWARD A. JOOS ◽  
SAMUEL W. CLAUSEN
Keyword(s):  
Serum Potassium ◽  
Whole Blood ◽  
Exchange Transfusion ◽  
Plasma Potassium ◽  
Cumulative Effects ◽  
Potassium Content ◽  
Prolonged Storage ◽  
Elevated Plasma ◽  
Serum Electrolyte ◽  
Transfusion Therapy

Alterations in concentrations of serum potassium are reported in infants studied during exchange transfusion therapy for erythroblastosis. The elevated plasma potassium content of citrated whole blood following prolonged storage may produce hyperkaliemia in some infants. The authors have suggested that hypocalcemia and hyperkaliemia may coexist in some of these babies during exchange transfusion. Certain implications regarding the cumulative effects of these two disturbances are discussed in relation to exchange transfusion.

Selective Lympholysis: A Unique Method Using the Double-Dye Technique.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 3884-3884
Author(s):  
William J. Owens ◽  
Thomas C. Cesario ◽  
Edward Shanbrom
Keyword(s):  
Methylene Blue ◽  
Whole Blood ◽  
Mononuclear Cells ◽  
Cell Types ◽  
The Novel ◽  
Treated Sample ◽  
Unique Method ◽  
Series Of Experiments ◽  
Lymphocytic Cell Line

Abstract Many methods are utilized to destroy mononuclear cells (primarily lymphocytes) either for neoplasm or immunosuppression. Neither radiation nor chemotherapy are truly selective or completely successful. The concept of “Double-Dye” treatment of blood for transfusion has been developed in order to inactivate parasites, bacteria and viruses (J Thromb Haemost2003; 1 Supplement 1 July: P1114). In recent studies, it has been observed that this same “Double-Dye” concept presents the possibility of very selectively eliminating lymphocytes (mononuclear cells) without affecting neutrophils in whole blood. To demonstrate the selectivity of dyes for lymphocytic/mononuclear cell types, two sets of experiments were performed. In the first, 0.3% (w/v) of the “Double Dye’ solution was added to several normal citrated whole blood samples to assess the effect on normal cells, compared to an untreated control. At 24hours post treatment, the lymphocyte count in the treated sample had dropped more than 80%, while little effect on neutrophils was noted. The control counts showed little change for either lymphocytes or neutrophils. Table 1. Lymphocyte reduction by 0.3%(w/v) Double-dye solution (units: cells/mm3). 0hr Control 0.3% Dye WBC 6850 ± 560 6920 ± 630 Neut 5030 ± 375 5100 ± 420 Lymph 1485 ± 220 1490 ± 265 24hr Control 0.3% Dye WBC 6700 ± 480 5300 ± 505 Neut 5025 ± 360 5025 ± 420 Lymph 1474 ± 240 275 ± 75 In the second series of experiments, 0.3%(w/v) “Double-Dye” solution or 0.15%(w/v) Crystal Violet or 0.15%(w/v) Methylene Blue were added to two T-cell leukemia lines (Jurkat, L1210), with a non-malignant, non-lymphocytic cell line (WISH) for the control. The combination of dyes showed the most potent activity against the lymphocytic lines, while the control was virtually unaffected. Table 2: Viability of cell lines after 24 hour exposure to dye solutions. Jurkat L1210 WISH Control 100% ± 2% 100% ± 4% 99% ± 2% 0.3% Double Dye 37% ± 5% 12% ± 4% 88% ± 9% 0.15% Meth. Blue 58% ± 12% 52% ± 9% 100% ± 3% 0.15% Cr. Violet 90% ± 12% 92% ± 10% 99% ± 4% The novel use of these dyes reported here coincided with the recent interest in utilizing methylene blue to increase transfusion safety, but recognizing that the concurrent need to photoactivate was too toxic to certain proteins and didn’t inactivate all pathogens (Transfusion2003; 43(9): 1238–47). Studies investigating the in-vivo efficacy of these novel immunosuppressive and chemotherapeutic methods are currently underway.

scholarly journals Delayed processing of blood samples impairs the accuracy of mRNA-based biomarkers

2022 ◽  
Author(s):  
Chace Wilson ◽  
Nicolas Dias ◽  
Stefania Pancini ◽  
Vitor Mercadante ◽  
Fernando Biase
Keyword(s):  
Rna Sequencing ◽  
Whole Blood ◽  
Biomarker Discovery ◽  
White Blood Cells ◽  
Transcript Abundance ◽  
Prolonged Storage ◽  
Gene Transcript ◽  
Sequencing Data ◽  
Blood Samples ◽  
Whole Blood Samples

Background: The transcriptome of peripheral white blood cells (PWBCs) contains valuable physiological information, thus making them a prime biological sample for investigating mRNA-based biomarkers. However, prolonged storage of whole blood samples can alter gene transcript abundance in PWBCs, compromising the results of biomarker discovery. Here, we designed an experiment to interrogate the impacts of delayed processing of whole blood samples on gene transcript abundance in PWBCs. We hypothesized that storing blood samples for 24 hours at 4°C would cause RNA degradation resulting in altered transcriptome profiles. Results: We produced RNA-sequencing data for 30 samples collected from five estrus synchronized heifers (Bos taurus). We quantified transcript abundance for 12,414 protein-coding genes in PWBCs. Analysis of parameters of RNA quality revealed no statistically significant differences (P>0.05) between samples collected from the jugular vein and coccygeal vein, as well as among samples processed after one, three, six, or eight hours. However, samples processed after 24 hours of storage had a lower RNA integrity number value (P=0.03) in comparison to those processed after one hour of storage. Next, we analyzed RNA-sequencing data between samples using those processed after one hour of storage as the baseline for comparison. Interestingly, evaluation of 3/5 prime bias revealed no differences between genes with lower transcript abundance in samples stored for 24 hours relative to one hour. In addition, sequencing coverage of transcripts was similar between samples from the 24-hour and one-hour groups. We identified four and 515 genes with differential transcript abundance in samples processed after storage for eight and 24 hours, respectively, relative to samples processed after one hour. Conclusions: The PWBCs respond to prolonged cold storage by increasing genes related to active chromatin compaction which in turn reduces gene transcription. This alteration in transcriptome profiles can impair the accuracy of mRNA-based biomarkers. Therefore, blood samples collected for mRNA-based biomarker discovery should be refrigerated immediately and processed within six hours post sampling.

scholarly journals STUDIES ON THE MECHANISM OF THE PHOTOSENSITIZED INACTIVATION OF E. COLI AND REACTIVATION PHENOMENON

1952 ◽  
Vol 36 (2) ◽  
pp. 207-226 ◽  
Author(s):  
F. Heinmets ◽  
R. Vinegar ◽  
W. W. Taylor
Keyword(s):  
Methylene Blue ◽  
Wave Length ◽  
Action Spectrum ◽  
Prolonged Storage ◽  
Experimental Conditions ◽  
E Coli ◽  
Hydrogen Donors ◽  
Inactivation Process ◽  
Photosensitized Inactivation ◽  
Inactivation Of Bacteria

In order to find a more satisfactory interpretation of the phenomenon of photosensitized inactivation of bacteria, studies were performed under various experimental conditions on methylene blue and E. coli. In summary the findings are as follow:— 1. The dye is absorbed by the bacteria according to the Langmuir isotherm and can be removed by ionic substitutions; the dye binding to the bacteria is predominantly ionic; the dye-bacteria complex produces a new absorption peak in the 610 mµ wave length region, and the action spectrum corresponds to the spectral absorption of the dye-bacteria complex. 2. There is an optimum dye concentration range for the photosensitized inactivation. 3. Photosensitized inactivation of bacteria can take place both in the frozen and liquid states and the presence of oxygen is essential to the inactivation process. 4. Hydrogen peroxide, formed by reoxidation of the reduced methylene blue, does not inactivate bacteria. 5. Following the photosensitized inactivation, E. coli lose their ability to reduce the methylene blue in the presence of various hydrogen donors, suggesting that enzymes are involved in the inactivation process. 6. Bacteria inactivated by photosensitization can be reactivated by prolonged storage after irradiation; the recovery rate increases with increasing temperature (maximum 37°), and is also influenced by the presence of various hydrogen donors. In view of collected experimental data, the basic reaction mechanisms are analyzed in photosensitized inactivation. The first step of the reaction seems to be excitation of the dye-bacteria, or dye-bacteria oxygen complex, by a photon which produces an activated complex. In such a state, molecular oxygen is capable of producing an oxidizing reaction, which results in the inactivation of the bacteria. Some aspects of the detailed reactions taking place at the cell surface are discussed.

scholarly journals Stored Whole Blood Transfusion Initiate Serum Amyloid A Activation Monitored by Real-Time Dynamic Imaging

2020 ◽  
Author(s):  
Yulong Zhang ◽  
Zhengjun Wang ◽  
Lei Liu ◽  
Qianqian Zhou ◽  
Sujing Sun ◽  
...  
Keyword(s):  
Blood Transfusion ◽  
Acute Phase ◽  
Whole Blood ◽  
Blood Cells ◽  
Serum Amyloid A ◽  
Prolonged Storage ◽  
Phase Reaction ◽  
Storage Lesion ◽  
Amyloid A ◽  
End Stage

Abstract BACKGROUND: Transfusion of stored whole blood (SWB) is increasingly routine practice to resuscitate severe traumatic hemorrhage patients in military operations and civilian emergency centers. It has been well established that transfusion of red blood cells (RBCs) after prolonged storage exerts harmful effects that are mainly mediated by inflammation. Whether the storage lesion that related to inflammation will happen in SWB remains unclear.METHODS: A hepatocyte SAA (serum amyloid A) specific reporter mouse that facilitated non-invasive imaging of hepatocyte SAA expression was used for the evaluation of acute inflammation and acute-phase reaction after the transfusion of SWB or components separated from end-stage whole blood. Donor C57BL/6 mouse whole blood was used to model an allogeneic transfusion in Balb/C mouse recipients. RESULTS: End-stage whole blood (14 days of storage) transfusion induced the most significant SAA expression, while 10-day-storage evoked much weaker signal compared to their fresh and 5-day-storage counterparts. It was RBCs rather than white blood cells and plasma-containing platelets that should be responsible for the systemic inflammatory and SAA activation during end-stage whole blood transfusion. Circulatory and hepatic pro-inflammatory cytokines induced the SAA expression in hepatocyte. The macrophage M1 polarization aroused by SWB activated SAA through nuclear transcription factor NF-κB. CONCLUSION: Storage lesion will also happen during the storage of whole blood, which is related to the change of RBCs after prolonged storage. The side effect induced by systemic inflammation and acute-phase reaction should be taken into consideration before resuscitation by long-term storage whole blood transfusion.

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