scholarly journals Effects of preservation duration at 4 °C on the quality of RNA in rabbit blood specimens

PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e8940
Author(s):  
Jiaojiao Song ◽  
Junmei Zhou
Keyword(s):  
Whole Blood ◽  
Rna Degradation ◽  
High Quality ◽  
Multicenter Studies ◽  
Rna Quality ◽  
Rabbit Blood ◽  
Significant Change ◽  
Blood Specimens ◽  
Storage Facilities

A prolonged preservation duration of blood specimens at 4 °C may occur due to the distance from collection points to storage facilities in many biobanks, especially for multicenter studies. This could lead to RNA degradation, affecting downstream analyses. However, effects of preservation durations at 4 °C on RNA quality in blood specimens need to be studied. We collected rabbit blood using EDTA tubes and stored them at 4 °C for different preservation durations. Then, we examined the quality of RNA from whole blood and leukocytes isolated from rabbit blood. Our results show that the purity of whole blood RNA and leukocyte RNA does not indicate significant change after rabbit blood is stored at 4 °C for different preservation durations (from 1 h to 7 days). The integrity of leukocyte RNA indicates the same result as above, but the integrity of whole blood RNA is significantly decreased after rabbit blood is stored at 4 °C for over 3 days. Moreover, expression of SMAD7, MKI67, FOS, TGFβ1 and HIF1α of whole blood RNA and leukocyte RNA remains basically stable, but PCNA expression of whole blood RNA or leukocyte RNA is significantly decreased after rabbit blood is stored at 4 °C for over 24 h or 7 days. Therefore, these results suggest that high-quality RNA is obtained from the fresher blood specimens and if blood specimens are stored for over 3 days at 4 °C, the quality of leukocyte RNA is more stable and of better quality than that of whole blood RNA.

scholarly journals Evaluating Alteration of Blood Transcriptome Affected by Storage Conditions Using RNA Sequencing

2020 ◽  
Author(s):  
Eun Jung Koh ◽  
So Yeon Yu ◽  
Seung Jun Kim ◽  
Eun-Il Lee ◽  
Seung Yong Hwang
Keyword(s):  
Gene Expression ◽  
Rna Sequencing ◽  
Whole Blood ◽  
Expression Patterns ◽  
Rna Degradation ◽  
Biological Research ◽  
P Value ◽  
Sample Handling ◽  
Rna Quality

Abstract BackgroundWhole blood is one of the most widely utilized human samples in biological research and is useful for analysing the mechanisms of diverse bio-molecular phenomena. However, owing to its fluidic properties, whole blood is relatively unstable in the frozen state compared to other biopsy samples. Because RNA is structurally unstable, sample damage can severely affect RNA quality, thereby reducing its usability. This study aimed to assess the quality of RNA prepared from blood stored at different temperatures and times prior to freezing, as well as the effect of freezer storage time. ResultsThe quality of the RNA derived from different blood samples was assessed by determining the RNA integrity number and RNA sequencing to identify genes (|fold-change (FC)| > 1.5, p-value < 0.05, false discovery rate (FDR) < 0.05) that were differentially expressed between the differently prepared RNA samples. We found that improper sample handling critically influenced both RNA quality and gene expression patterns. In particular, storing blood at room temperature over 12 h before freezing led to RNA degradation. Differential gene expression analysis revealed that expression of the CXCR1 gene was substantially reduced when using impaired RNA. ConclusionsThis study emphasizes the importance of proper sample management for obtaining reliable downstream application outcomes and suggests the CXCR1 gene as a candidate screening marker for RNA damage caused by improper sample handling.

scholarly journals Optimized protocol for the extraction of RNA and DNA from frozen whole blood sample stored in a single EDTA tube

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Hirotaka Yamagata ◽  
Ayumi Kobayashi ◽  
Ryouichi Tsunedomi ◽  
Tomoe Seki ◽  
Masaaki Kobayashi ◽  
...  
Keyword(s):  
Whole Blood ◽  
Ethylenediaminetetraacetic Acid ◽  
Rna Extraction ◽  
Basic Research ◽  
Rna Degradation ◽  
Disodium Salt ◽  
Water Bath ◽  
High Quality ◽  
Blood Samples ◽  
Rna And Dna

AbstractCryopreservation of whole blood is useful for DNA collection, and clinical and basic research. Blood samples in ethylenediaminetetraacetic acid disodium salt (EDTA) tubes stored at − 80 °C are suitable for DNA extraction, but not for high-quality RNA extraction. Herein, a new methodology for high-quality RNA extraction from human blood samples is described. Quickly thawing frozen whole blood on aluminum blocks at room temperature could minimize RNA degradation, and improve RNA yield and quality compared with thawing the samples in a 37 °C water bath. Furthermore, the use of the NucleoSpin RNA kit increased RNA yield by fivefold compared with the PAXgene Blood RNA Kit. Thawing blood samples on aluminum blocks significantly increased the DNA yield by ~ 20% compared with thawing in a 37 °C water bath or on ice. Moreover, by thawing on aluminum blocks and using the NucleoSpin RNA and QIAamp DNA Blood kits, the extraction of RNA and DNA of sufficient quality and quantity was achieved from frozen EDTA whole blood samples that were stored for up to 8.5 years. Thus, extracting RNA from frozen whole blood in EDTA tubes after long-term storage is feasible. These findings may help advance gene expression analysis, as well as biomarker research for various diseases.

Assessment of platelet activation in several different anticoagulants by the Advia 120 Hematology System, fluorescence flow cytometry, and electron microscopy

2003 ◽  
Vol 90 (11) ◽  
pp. 940-948 ◽  
Author(s):  
Sabrinah Chapman ◽  
Mariette Lépine ◽  
David Okrongly ◽  
Nuria Pujol-Moix ◽  
Angel Hernández ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Platelet Activation ◽  
Whole Blood ◽  
Sodium Citrate ◽  
Ethylenediaminetetraacetic Acid ◽  
Density Parameter ◽  
Flow Cytometric ◽  
Significant Change ◽  
Blood Specimens ◽  
Advia 120

Summary In vivoplatelet activation results are often confounded by activation induced in vitroduring the preparative procedures. We measured ex vivo(basal) and in vitro(thrombin-induced) platelet activation in sodium citrate, ethylenediaminetetraacetic acid (EDTA), and Citrate Theophylline Dipyridamole Adenosine (CTAD) whole blood specimens. Determinations were made by measurements of platelet density (mean platelet component: MPC concentration) on the Advia 120 Hematology System. The MPC has been previously shown to correlate with a fluorescence flow cytometric method, also determined in this study, using the surface expression of CD62P. Moreover, platelet shape and structure changes in EDTA and CTAD anticoagulated whole blood specimens were characterized by transmission electron microscopy (TEM). Observations made using the Advia 120 Hematology System platelet density parameter, MPC, in the absence of thrombin were 25.7 ± 0.9 g/dl, 27.9 ± 0.9 g/dl and 24.8 ± 1.2 g/dl in sodium citrate, EDTA and CTAD whole blood specimens, respectively. Addition of thrombin induced a significant change in platelet MPC for sodium citrate (21.9 ± 1.9 g/dl; p<0.0001) and EDTA (23.2 ± 0.9 g/dl; p<0.0001) whole blood specimens. In contrast, thrombin had no effect on MPC measured in whole blood taken into CTAD tubes. In vitrofluorescence flow cytometric platelet activation experiments measuring the percentage of platelets expressing anti-CD62P showed increase in sodium citrate specimens from 9.2 ± 7.0 to 55.5 ± 23.1 % (p<0.0001) and in EDTA specimens from 1.9 ± 1.7 to 64.6 ± 12.4 % (p<0.0001) after addition of thrombin. However, in blood taken into CTAD tubes, there was no significant change. Studies on platelets isolated from whole blood in CTAD showed activation by thrombin indicating that platelets in CTAD, while protected in its presence remained functional upon its removal. When observed by TEM over time, platelets in EDTA appear more activated and contain fewer granules than platelets in CTAD. We conclude that CTAD demonstrates in vitroplatelet activation inhibition and may be useful in stabilizing ex vivoplatelet activation. The novel platelet activation parameter, MPC, measured by an automated routine hematology system, using customized proprietary software, may be used in conjunction with CTAD, a stabilizing anticoagulant, to measure the ex vivoplatelet activation state in whole blood specimens. TEM studies verify shape modifications and simultaneous retention of intracellular granules at early post-venipuncture time periods in CTAD specimens.

Influence of technology parameters of free-cutting steel continuous casting on the billet internal structure

2020 ◽  
Vol 76 (2) ◽  
pp. 139-142
Author(s):  
A. T. Kunakbaeva ◽  
A. M. Stolyarov ◽  
M. V. Potapova
Keyword(s):  
Regression Analysis ◽  
Continuous Casting ◽  
Internal Structure ◽  
Sulfur Content ◽  
Manganese Content ◽  
High Quality ◽  
Correlation And Regression Analysis ◽  
Cutting Steel ◽  
Continuously Cast

Free-cutting steel gains specific working properties thanks to the high content of sulfur and phosphorus. These elements, especially sulfur, have a rather high tendency to segregation. Therefore, segregation defects in free-cutting steel continuously cast billets can be significantly developed. The aim of the work was to study the influence of the chemical composition of freecutting steel and casting technological parameters on the quality of the macrostructure of continuously cast billets. A metallographic assessment of the internal structure of cast metal made of free-cutting steel and data processing by application of correlation and regression analysis were the research methods. The array of production data of 43 heats of free-cutting steel of grade A12 was studied. Steel casting on a five-strand radial type continuous casting machine was carried out by various methods of metal pouring from tundish into the molds. Metal of 19 heats was poured with an open stream, and 24 heats – by a closed stream through submerged nozzles with a vertical hole. High-quality billets had a cross-sectional size of 150×150 mm. The macrostructure of high-quality square billets made of free-cutting steel of A12 grade is characterized by the presence of central porosity, axial segregation and peripheral point contamination, the degree of development of which was in the range from 1.5 to 2.0 points, segregation cracks and strips – about 1.0 points. In the course of casting with an open stream, almost all of these defects are more developed comparing with the casting by a closed stream. As a result of correlation and regression analysis, linear dependences of the development degree of segregation cracks and strips both axial and angular on the sulfur content in steel and on the ratio of manganese content to sulfur content were established. The degree of these defects development increases with growing of sulfur content in steel of A12 grade. These defects had especially strong development when sulfur content in steel was of more than 0.10%. To improve the quality of cast metal, it is necessary to have the ratio of the manganese content to the sulfur content in the metal more than eight.

Experience of clinical testing of personal telemedicine system ‘Obereg’ for remote monitoring of patients in intensive care units

2020 ◽  
pp. 52-58 ◽  
Author(s):  
A. A. Eryomenko ◽  
N. V. Rostunova ◽  
S. A. Budagyan ◽  
V. V. Stets
Keyword(s):  
Intensive Care ◽  
Intensive Care Units ◽  
Remote Monitoring ◽  
Clinical Testing ◽  
Transportation Of Patients ◽  
High Quality ◽  
Telemedicine System ◽  
Accuracy Of Measurements ◽  
Hospital Equipment

The experience of clinical testing of the personal telemedicine system ‘Obereg’ for remote monitoring of patients at the intensive care units of leading Russian clinics is described. The high quality of communication with the remote receiving devices of doctors, the accuracy of measurements, resistance to interference from various hospital equipment and the absence of its own impact on such equipment were confirmed. There are significant advantages compared to stationary patient monitors, in particular, for intra and out-of-hospital transportation of patients.

METHODOLOGICAL ASPECTS OF VISUALIZATION OF CORONARY ARTERIES WITH TRANSTHORACIC ECHOCARDIOGRAPHY

2018 ◽  
pp. 26-35
Author(s):  
Z. A. Agaeva ◽  
K. B. Baghdasaryan
Keyword(s):  
Blood Flow ◽  
Coronary Blood Flow ◽  
Coronary Arteries ◽  
Transthoracic Echocardiography ◽  
Second Harmonic ◽  
Anatomic Reconstruction ◽  
High Quality ◽  
Acoustic Window ◽  
Methodological Aspects

The transthoracic echocardiography made by multifrequency probes with support of the mode of the second harmonic imaging, is a competitive method for visualization of the main coronary arteries and allows to estimate coronary blood flow with high quality. Of course, the method has considerable restrictions, most important of which is the low spatial resolution of a method, due to small acoustic window. Because of this the transthoracic visualization of coronary arteries perhaps will not become the leading method of anatomic reconstruction of separately taken coronary artery and especially all coronary arteries system. However uniqueness and indisputable advantage of this method is an opportunity to noninvasively estimate a coronary blood flow both once, and in dynamics.

Estimates on poverty in Russia and the additional measures to reduce it at the regional level

2020 ◽  
Vol 18 (4) ◽  
pp. 739-752 ◽  
Author(s):  
R.M. Sadykov
Keyword(s):  
Poverty Rate ◽  
Regional Level ◽  
Living Standards ◽  
Standard Of Living ◽  
High Quality ◽  
Social Standard ◽  
Republic Of Bashkortostan ◽  
The Republic ◽  
Sociological Studies

Subject. This article deals with the issues of social justice and a high quality of life, creating favorable economic and social conditions. Objectives. The article aims to assess the rate and changes in poverty in Russia and the Republic of Bashkortostan and develop complementary measures to reduce it. Methods. For the study, I used the methods of logical, comparative, economic and statistical analyses, the results of sociological studies, and official statistics. Results. The article highlights additional measures to reduce poverty in the region, including the establishment of a minimum social standard of living in each particular region that determines the poverty rate. Conclusions. Various factors, such as economic sanctions, economic slowdowns, territorial and regional imbalances, lead to living standards decline and poverty rise.

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