scholarly journals Effects of Blood Sample Age at Time of Separation on Measured Cytokine Concentrations in Human Plasma

2010 ◽  
Vol 18 (2) ◽  
pp. 318-326 ◽  
Author(s):  
Rachael P. Jackman ◽  
Garth H. Utter ◽  
John W. Heitman ◽  
Dale F. Hirschkorn ◽  
Jacqueline P. Law ◽  
...  
Keyword(s):  
Whole Blood ◽  
Peripheral Blood ◽  
Blood Protein ◽  
Blood Collection ◽  
Trauma Patients ◽  
Immune Mediators ◽  
Blood Cytokines ◽  
Wide Range ◽  
Significant Change ◽  
Sample Age

ABSTRACTMeasurement of peripheral blood cytokines and other immunomodulatory proteins is a useful and popular tool for assessing human immune responses to a wide range of assaults. A common challenge in this work is obtaining fresh, high-quality samples and limiting the time between blood collection and the separation of plasma or serum from cells. In this study we sought to determine the effect of sample age at the time of processing on the measured levels of 41 soluble immune mediators. Two cohorts were examined: healthy lab donors and trauma patients, who have significant immune perturbation. Whole-blood samples were aliquoted, and plasma was isolated, at days 0, 1, 2, and 3 after collection. Multiplexing techniques were used to measure protein concentrations, and general estimating equations were used to determine if there was a significant change over time. Over the 3-day period examined, only 15 of the 41 proteins showed no significant change in either cohort. Among the remaining proteins both increases and decreases were observed, with changes ranging from 2.4% per day to 325% per day. Proteins with significant changes in one cohort did not always show significant changes in the other group. These results support the need to separate plasma or serum from whole blood as quickly as possible and/or to standardize the length of time to processing within a given study of peripheral blood protein concentrations. When this is not possible, care should be taken to account for differences due to sample age.

scholarly journals Quantitative Reverse Transcription-PCR Measurement of Thyroglobulin mRNA in Peripheral Blood of Healthy Subjects

1999 ◽  
Vol 45 (6) ◽  
pp. 785-789 ◽  
Author(s):  
Susan T Wingo ◽  
Matthew D Ringel ◽  
Jeffrey S Anderson ◽  
Aneeta D Patel ◽  
Yvonne D Lukes ◽  
...  
Keyword(s):  
Thyroid Cancer ◽  
Reverse Transcription ◽  
Whole Blood ◽  
Peripheral Blood ◽  
Thyroid Disease ◽  
Healthy Subjects ◽  
Extraction Technique ◽  
Blood Collection ◽  
Rt Pcr ◽  
Total Rna

Abstract Background: Thyroglobulin mRNA can be detected qualitatively in the peripheral blood of patients with metastatic thyroid cancer, thyroid cancer patients with residual thyroid bed uptake, and individuals with no known thyroid disease with intact thyroid glands by use of a lengthy, highly sensitive extraction technique. To improve and broaden the clinical usefulness of this assay, we developed a quantitative reverse transcription (RT)-PCR assay for thyroglobulin mRNA, using RNA recovered from whole blood with a simplified extraction technique. Methods: Whole blood was drawn from 32 healthy subjects in standard EDTA blood collection tubes. Total RNA was extracted from whole blood, using the PUREscript RNA Isolation Kit. RT-PCR using intron-spanning primers was used to quantitatively amplify thyroglobulin mRNA, using the ABI PRISM 7700 Sequence Detection System with a fluorescent-labeled, thyroglobulin-specific oligonucleotide probe. Thyroid RNA calibration curves were created using total RNA recovered from a single nondiseased thyroid gland. Results: Qualitative RT-PCR demonstrated the presence of thyroglobulin mRNA in the whole blood sample of each healthy subject. The mean concentration of thyroglobulin mRNA detected in these subjects was 433 ± 69 ng of total thyroid RNA per liter of whole blood (range, 26–1502 ng/L). Overall assay imprecision (CV) was 24% for five samples analyzed 10 times each in separate analytical runs on different days. Conclusions: Thyroglobulin mRNA can be accurately detected and quantified in peripheral blood from healthy subjects. This new quantitative technique may improve the clinical utility of circulating thyroglobulin mRNA detection in patients with thyroid disease.

scholarly journals homeRNA: A self-sampling kit for the collection of peripheral blood and stabilization of RNA

2021 ◽  
Author(s):  
Amanda J. Haack ◽  
Fang Yun Lim ◽  
Dakota S. Kennedy ◽  
John H. Day ◽  
Karen N. Adams ◽  
...  
Keyword(s):  
Whole Blood ◽  
Peripheral Blood ◽  
Rna Degradation ◽  
Blood Collection ◽  
Rna Integrity ◽  
Rna Integrity Number ◽  
Collection Device ◽  
Location Parameters ◽  
Rna Stabilization ◽  
Gene Panels

ABSTRACTGene expression analysis (e.g., targeted small gene panels, transcriptomics) from whole blood can elucidate mechanisms of immune function and aid in the discovery of biomarkers. Conventional in-clinic venipuncture offers only a small snapshot of our broad immune landscape as immune responses may occur outside of the time and location parameters available for conventional venipuncture. A self-operated method that enables flexible sampling of liquid whole blood coupled with an immediate stabilization of cellular RNA is instrumental in facilitating capture and preservation of acute or transient immune fluxes. To this end, we developed homeRNA: a kit that allows for self-collection of peripheral blood (∼0.5 mL) and immediate stabilization of cellular RNA, using the Tasso-SST™ blood collection device paired with a specially designed stabilizer tube containing RNAlater™. To assess the usability and feasibility of homeRNA for self-collection and stabilization of whole blood RNA, we conducted a pilot study (n = 41 participants) where we sent homeRNA to participants aged 21-69, located across 10 US states (94% successful blood collections, n = 51). Among participants who successfully collected blood, 91% reported no or minimal pain/discomfort using the kit (n = 35), and 77% reported easy or somewhat easy stabilization protocol. Total RNA yield from the stabilized samples ranged between 0.24 µg and 5.99 µg (mean = 1.65 µg), while RNA Integrity Number (RIN) values were above 7.0 (mean = 7.9), indicating limited RNA degradation. Results from this study demonstrate the self-collection and RNA stabilization of whole blood with homeRNA by participants themselves, in their own home.

scholarly journals Expanding the range of sausage products of special purpose

2020 ◽  
Vol 22 (93) ◽  
pp. 72-76
Author(s):  
O. Haschuk ◽  
O. Moskalyuk ◽  
I. Simonova
Keyword(s):  
Amino Acids ◽  
Whole Blood ◽  
Binding Capacity ◽  
Control Sample ◽  
Partial Replacement ◽  
Blood Protein ◽  
Fat Emulsion ◽  
Organoleptic Evaluation ◽  
Wide Range ◽  
Minced Meat

The article presents the results of research of minced meat from turkey meat with the addition of whole blood in order to expand the range of special purpose products. The presence of a wide range of physiologically active substances in meat raw materials determines its special properties. In scientific work was conducted influence of products of blood processing on qualitative indicators of sausages of health-improving purposes. The use of blood protein Globin Vepro Gel 95 HV in the composition of protein-fat emulsion (in the amount of 40 % by weight of minced meat) and whole blood to increase the iron content in the product for the prevention and treatment of anemia. During the organoleptic evaluation of meat systems, it was found that partial replacement of raw meat in minced meat with whole blood has a beneficial effect on the color of the product. But with the increase in the amount of blood there is a deterioration of organoleptic parameters. The addition to the minced meat of a protein-fat emulsion based on globin protein and the addition of whole blood in appropriate proportions creates the conditions for moisture binding. However, with the addition of 10 % of blood, there is a decrease in moisture binding capacity by 0.3 %. According to the results of studies of the content of essential amino acids, it was found that the experimental samples of sausages are characterized by the content of complete proteins. In particular, the sample № 3, which has the best functional and technological indicators has a higher rate of Lysine, Leucine, Methionine + Cysteine, Valine compared to the control sample. The dominant amino acids are Lysine, Threonine, Leucine, Phenylalanine and Tyrosine, Limiting – Methionine, Cystine, Valine, Isoleucine. For sample № 3, the value of the utilization coefficient of the protein is 0.52, the biological value is 61.12 %.

Measurement of glycosylated whole-blood protein for assessing glucose control in diabetes: collection and storage of capillary blood on filter paper.

1985 ◽  
Vol 31 (2) ◽  
pp. 213-216 ◽  
Author(s):  
R R Little ◽  
H M Wiedmeyer ◽  
J D England ◽  
W C Knowler ◽  
D E Goldstein
Keyword(s):  
Filter Paper ◽  
Whole Blood ◽  
Glycosylated Hemoglobin ◽  
Colorimetric Method ◽  
Capillary Blood ◽  
Blood Protein ◽  
Blood Collection ◽  
Small Contribution ◽  
Blood Proteins ◽  
Highly Correlated

Abstract We present data on the use of filter-paper blood collection for measurement of glycosylated whole-blood proteins (gWB) (hemoglobin and plasma proteins). A capillary blood sample, obtained by fingerprick, is spotted directly onto filter paper (Schleicher & Schuell 903). The blood spot is washed briefly with alcohol (ethanol or isopropanol) to remove free glucose and dried before shipment to the laboratory. In the laboratory, the blood is eluted from the paper and analyzed for gWB by a colorimetric method. The gWB is primarily a measure of glycosylated hemoglobin (gHb) with a small contribution from glycosylated plasma protein. Concentrations of gWB and gHb are highly correlated (r = 0.91). The filter-paper method offers advantages over currently available methods for quantifying gHb and may be particularly useful in screening for diabetes and for assessing glycemic control in patients from remote areas.

Stability of antioxidant vitamins in whole human blood during overnight storage at 4°C and frozen storage up to 6 months

2018 ◽  
Vol 88 (3-4) ◽  
pp. 151-157 ◽  
Author(s):  
Scott W. Leonard ◽  
Gerd Bobe ◽  
Maret G. Traber
Keyword(s):  
Ascorbic Acid ◽  
Whole Blood ◽  
Healthy Subjects ◽  
Frozen Storage ◽  
Blood Collection ◽  
Plasma Samples ◽  
Optimal Conditions ◽  
Plasma Ascorbic Acid ◽  
Blood Samples ◽  
Sample Handling

Abstract. To determine optimal conditions for blood collection during clinical trials, where sample handling logistics might preclude prompt separation of erythrocytes from plasma, healthy subjects (n=8, 6 M/2F) were recruited and non-fasting blood samples were collected into tubes containing different anticoagulants (ethylenediaminetetra-acetic acid (EDTA), Li-heparin or Na-heparin). We hypothesized that heparin, but not EDTA, would effectively protect plasma tocopherols, ascorbic acid, and vitamin E catabolites (α- and γ-CEHC) from oxidative damage. To test this hypothesis, one set of tubes was processed immediately and plasma samples were stored at −80°C, while the other set was stored at 4°C and processed the following morning (~30 hours) and analyzed, or the samples were analyzed after 6 months of storage. Plasma ascorbic acid, as measured using HPLC with electrochemical detection (LC-ECD) decreased by 75% with overnight storage using EDTA as an anticoagulant, but was unchanged when heparin was used. Neither time prior to processing, nor anticoagulant, had any significant effects upon plasma α- or γ-tocopherols or α- or γ-CEHC concentrations. α- and γ-tocopherol concentrations remained unchanged after 6 months of storage at −80°C, when measured using either LC-ECD or LC/mass spectrometry. Thus, refrigeration of whole blood at 4°C overnight does not change plasma α- or γ-tocopherol concentrations or their catabolites. Ascorbic acid is unstable in whole blood when EDTA is used as an anticoagulant, but when whole blood is collected with heparin, it can be stored overnight and subsequently processed.

Status of humoral immune factors in peripheral blood of fronto-orbital trauma patients late after surgery with the use of biocomposite

2019 ◽  
Vol 82 (5) ◽  
pp. 37-41
Author(s):  
О. Bondarchuk ◽  
◽  
V. Kishchuk ◽  
О. Melnykov ◽  
М. Tymchenko ◽  
...  
Keyword(s):  
Peripheral Blood ◽  
Trauma Patients ◽  
Orbital Trauma ◽  
Humoral Immune ◽  
Immune Factors

scholarly journals An Evaluation of Morphological Changes and Deformability of Suspended Red Blood Cells Prepared Using Whole Blood with Different Hemoglobin Levels of Tibetans

2021 ◽  
pp. 1-10
Author(s):  
Rui Zhong ◽  
Dingding Han ◽  
Xiaodong Wu ◽  
Hong Wang ◽  
Wanjing Li ◽  
...  
Keyword(s):  
Red Blood Cells ◽  
Whole Blood ◽  
Blood Cells ◽  
Morphological Changes ◽  
Osmotic Fragility ◽  
Spherical Shape ◽  
Hypoxic Environment ◽  
Wide Range ◽  
Group A ◽  
Cell Membrane Protein

Background: The hypoxic environment stimulates the human body to increase the levels of hemoglobin (HGB) and hematocrit and the number of red blood cells. Such enhancements have individual differences, leading to a wide range of HGB in Tibetans’ whole blood (WB). Study Design: WB of male Tibetans was divided into 3 groups according to different HGB (i.e., A: >120 but ≤185 g/L, B: >185 but ≤210 g/L, and C: >210 g/L). Suspended red blood cells (SRBC) processed by collected WB and stored in standard conditions were examined aseptically on days 1, 14, 21, and 35 after storage. The routine biochemical indexes, deformability, cell morphology, and membrane proteins were tested. Results: Mean corpuscular volume, adenosine triphosphate, pH, and deformability were not different in group A vs. those in storage (p > 0.05). The increased rate of irreversible morphology of red blood cells was different among the 3 groups, but there was no difference in the percentage of red blood cells with an irreversible morphology after 35 days of storage. Group C performed better in terms of osmotic fragility and showed a lower rigid index than group A. Furthermore, SDS-PAGE revealed similar cross-linking degrees of cell membrane protein but the band 3 protein of group C seemed to experience weaker clustering than that of group A as detected by Western Blot analysis after 35 days of storage. Conclusions: There was no difference in deformability or morphological changes in the 3 groups over the 35 days of storage. High HGB levels of plateau SRBC did not accelerate the RBC change from a biconcave disc into a spherical shape and it did not cause a reduction in deformability during 35 days of preservation in bank conditions.

scholarly journals Assessment of Donkey (Equus asinus africanus) Whole Blood Stored in CPDA-1 and CPD/SAG-M Blood Bags

Biology ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 133
Author(s):  
Isabella Oliveira Barros ◽  
Rejane Santos Sousa ◽  
Marcondes Dias Tavares ◽  
Renato Otaviano Rêgo ◽  
Paulo Ricardo Firmino ◽  
...  
Keyword(s):  
Blood Cell ◽  
Cell Count ◽  
Whole Blood ◽  
Blood Collection ◽  
Blood Gas ◽  
Blood Cell Count ◽  
Animal Blood ◽  
Equus Asinus ◽  
Citrate Phosphate Dextrose ◽  
Citrate Phosphate

Hemotherapy using whole blood and its components is being increasingly used in veterinary therapy. Since it is important to store animal blood while maintaining acceptable hematological, blood gas, and biochemical characteristics, increasing our knowledge of available technologies for strategic blood storage is imperative. Thus, we aimed to assess the hematological, blood gas, and biochemical changes in donkey whole blood using blood bags with two different types of storage agents. Eight adult healthy male donkeys were used; 900 mL of blood was collected from each, with 450 mL stored in citrate-phosphate-dextrose and adenine bags (CPDA-1) and 450 mL stored in bags containing citrate-phosphate-dextrose, adenine, mannitol, and sodium chloride (CPD/SAG-M). Both bags were kept refrigerated between 1 and 6 °C for 42 days. Blood samples were removed from the bags eight times (T): T0 (immediately after blood collection), T1, T3, T7, T14, T21, T35, and T42 (1, 3, 7, 14, 21, 35 and 42 days after storage). Hematological, blood gas, biochemical, and microbiological parameters were assessed. The CPDA-1 bags had a higher packed cell volume when compared to CPD/ SAG-M. The red blood cell count reduced by around 19% in both the bags due to hemolysis, which was confirmed by an increase in plasma hemoglobin. The white blood cell count; pH; concentrations of glucose, sodium, bicarbonate, and 2,3 diphosphoglycerate were reduced in both bags. Meanwhile, pO2, pCO2, lactate dehydrogenase, and levels of potassium increased in the CPDA-1 and CPD/SAG-M bags. Blood bags were efficient for the storage of donkey blood for up to 42 days.

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