scholarly journals A Comprehensive Comparison of DNA Extraction Using Fresh and Stored Bloods in Molecular Hematology Diagnostic Study

2018 ◽  
Vol 17 (3) ◽  
pp. 424-432
Author(s):  
Siti Nazihahasma Hassan ◽  
Suharni Mohamad ◽  
Rosline Hassan ◽  
Selamah Ghazali ◽  
Wan Suriana Wan Ab Rahman
Keyword(s):  
Medical Science ◽  
Pcr Amplification ◽  
Diagnostic Study ◽  
Fresh Blood ◽  
Blood Samples ◽  
Acceptable Alternative ◽  
Significant Difference ◽  
Comprehensive Comparison ◽  
One Year ◽  
Stored Blood

Objective: Blood is the main source of DNA in molecular biology. It provides a high DNA quality and quantity. In this study, we compare the quality and quantity of DNA isolated from stored blood that has been kept at -40°C for one-year to that of fresh blood.Materials and Methods: Twelve fresh and stored blood samples were randomly selected for this study. Nucleo Spin® Blood L kit was used to isolate the DNA from the samples. The integrity and intensity of DNA were examined through 1.6% agarose gel precast with SYBR® safe DNA stain. The DNA samples were further examined through PCR amplification and Sanger sequencing.Results: There was no significant difference in quality and quantity of isolated DNA from fresh blood and stored blood samples. The high intensity of an intact DNA band as well as the success in PCR amplification and sequencing are indicators of high DNA quality.Conclusion: Proper storage of patients’left-over whole blood sample at -40°C offers an acceptable alternative for DNA resources in molecular study.Bangladesh Journal of Medical Science Vol.17(3) 2018 p.424-432

Re-inventing the wheel: the use of autologous and fresh donor blood in cardiac surgery

Perfusion ◽  
1995 ◽  
Vol 10 (2) ◽  
pp. 93-99 ◽  
Author(s):  
Ian J Reece ◽  
Gerrard H Linley ◽  
Jitesh Tolia ◽  
Jayesh Sheth ◽  
Habib Al Tareif
Keyword(s):  
Intensive Care ◽  
Renal Dysfunction ◽  
Autologous Blood ◽  
Fresh Blood ◽  
Donor Blood ◽  
Chest Drainage ◽  
Short Supply ◽  
Significant Difference ◽  
Glucose 6 Phosphate Dehydrogenase ◽  
Stored Blood

From the opening of a new cardiac surgical programme in November 1992, autologous and fresh donor blood (FDB) were used rather than cold stored blood (CSB) wherever possible in patients undergoing operations involving the use of cardiopulmonary bypass (CPB). In the first 250 consecutive patients, autologous blood was used in 168 (67.2%), fresh blood was used in 188 (75.2%). A total of 740 units of fresh blood were obtained on the day of operation (mean 3.9 ± 1.6 units per patient able to supply donors; 4.9 ± 1.7 units in the 147 who received fresh blood) and 728 units of stored blood were used (mean 3.08 ± 1.84 units per patient where fresh blood was used; 6.2 ± 2.5 units in the 114 where no fresh blood was used). The use of autologous blood significantly reduced FDB and CSB requirements (p < 0.001), was associated with a shorter intensive care and total postoperative stay (p = 0.006 and p = 0.033 respectively), even though there were more urgent and emergency cases in this group (p = 0.009) and no significant difference in chest drainage. Coagulopathy developed in 41 patients (16.4%) and was significantly associated with bypass time (p = 0.0001) and preoperative renal dysfunction (p = 0.005), although not with advanced age, sex, redo operation, diabetes or glucose-6-phosphate dehydrogenase deficiency. Patients with coagulopathy had significantly more transfused blood and blood products (p = 0.0001) and longer intensive care and total postoperative stays (p = 0.0001). In terms of blood conservation, the use of autologous blood was of primary importance. Correction of renal dysfunction and keeping overall bypass time to a minimum reduced the incidence of coagulopathy and, therefore, blood use. The use of FDB in this series was not clearly associated with improved overall results, but did generate more blood than was actually used, a factor of extreme importance in situations where stored blood is always in short supply.

scholarly journals Biochemical responses, blood gas, oxidative stress and lipid peroxidation of goats transfused with fresh or stored whole homologous blood

2018 ◽  
Vol 38 (11) ◽  
pp. 2070-2079 ◽  
Author(s):  
Nayanna B.S. Fonseca ◽  
Jucélio S. Gameleira ◽  
Jerson M. Cavalcante ◽  
Francisco L.C. Oliveira ◽  
Clara S. Mori ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Lipid Peroxidation ◽  
Whole Blood ◽  
Total Protein ◽  
Gas Analysis ◽  
Blood Gas ◽  
Fresh Blood ◽  
Sod Activity ◽  
Blood Samples ◽  
Stored Blood

ABSTRACT: Blood transfusion is a therapeutic procedure of great importance for veterinary medicine, in spite of only few studies in the literature on hemotherapy in goats. We aimed to evaluate the biochemical, blood gas, oxidative stress and lipid peroxidation of goats submitted to homologous transfusion of fresh whole blood or stored for 15 and 35 days. Eighteen adult male goats were submitted to a single phlebotomy to remove 30% of the blood volume, and we transfused 20mL/kg of whole blood stored in CPDA-1 bags according to the experimental group, being: G0 composed goats who received fresh blood, G15 and G35 goats that received blood stored for 15 and 35 days, respectively. For the biochemical evaluation, blood gas, oxidative stress and lipid peroxidation, blood samples were collected at the following moments: before the induction of anemia (TC0); 6 hours after phlebotomy and before transfusion (TC1); 1, 6, 12, 24 and 96 hours after transfusion (T1, T6, T12, T24 and T96 respectively); 8, 16 and 32 days after transfusion (T8d, T16d and T32d respectively). Before transfusion, blood samples were also withdrawn from the bags for the same analyzes. Statistical analyzes were performed in the statistical program GRAPHPAD PRISM 5.0, adopting a significance level of 5%. The bags of blood stored for 15 and 35 days showed more biochemical changes, blood gas, oxidative stress and lipid peroxidation than fresh blood bags. As for the biochemical analysis, after the transfusion was observed an increase of the total protein, albumin, glucose and creatine kinase in the 3 groups, and elevation of total bilirubin, direct bilirubin, and urea in G15 and G35. The changes observed in the blood gas analysis had no clinical significance, as they were within the reference values for the species. The goats that received stored blood showed disorder in their antioxidant system through alteration of the SOD activity. In the analysis of lipid peroxidation no difference between the groups for the concentration of malondialdehyde was found. Thus, it can be concluded that transfusion of whole fresh stored blood in goats did not compromise the blood gases, lipid peroxidation and liver and renal functions of the transfused animals. In addition, the method was proved to be efficient to restore, among other components, the total protein and albumin. The transfusion, as performed in this study, proved to be safe for used in the clinical practice of goats.

scholarly journals Assessment of prevalence of sexually transmissible infections among voluntary blood donors and its comparison with replacement donors at Hassan Institute of Medical Science, Hassan, Karnataka, India

2018 ◽  
Vol 5 (4) ◽  
pp. 901
Author(s):  
Bindu C. B. ◽  
Lokesh H. C. ◽  
Purushotham R.
Keyword(s):  
Blood Donors ◽  
Medical Science ◽  
Blood Samples ◽  
Total Prevalence ◽  
Sexually Transmissible Infections ◽  
Significant Difference ◽  
B Virus ◽  
Hbv Prevalence ◽  
History Of ◽  
Acid Test

Background: Hepatitis B virus, Hepatitis C virus and Human immunodeficiency virus (HIV) are important STDs which can be transmissible to the recipients of blood transfusion. The aim of the present study is to study the seroprevalence of HIV, HBV and HCV infection in the blood among voluntary and replacement donors in HIMS Hassan during 2010 to 2012.Methods: A retrospective study was conducted at blood bank of HIMS, Hassan for the years 2010 to 2012. The donors with Hemoglobin>12gm% for both sexes, weight >50 kg, no history of chronic illness, hepatitis, high risk behaviours were included in the study. All the blood samples collected were screened for HIV, HBV and HCV using ELISA kits. All the blood samples were sent to NACO (national AIDS control organization) and subjected to NAT (nucleic acid test) for detection of antigens. Results compared for both voluntary and replacement donors.Results: Total of 10938 blood donors screened. Majority of the donors were males 95.8% (10484) and belonged to voluntary group 72.8% (7971). The total prevalence of STDS were 0.61% (67). The prevalence of HBV, HCV and HIV was 0.47% (51), 0.04% (4) and 0.11% (12) respectively. Prevalence of STDs was higher among voluntary donors 0.57% (62) compared to replacement donors 0.05 % (5). Statistically significant difference was observed in HBV prevalence in voluntary and replacement donors.Conclusions: Most common STDs in blood donors was HBV followed by HIV and HCV. STDs were mainly seen in voluntary donors compared to replacement donors. Majority of the donors were males.

scholarly journals Evaluation of Serum α-tocopherol and Anemia among Infertile Patients Attending Specialists Hospital, Sokoto

2019 ◽  
pp. 1-5
Author(s):  
F. U. Bunza ◽  
A. S. Mainasara ◽  
I. Z. Wasagu ◽  
I. Y. Maryam
Keyword(s):  
Average Period ◽  
Independence Test ◽  
Unprotected Sexual Intercourse ◽  
Blood Samples ◽  
Significant Difference ◽  
Student’S T ◽  
One Year ◽  
Avitaminosis E ◽  
Infertile Patients ◽  
Apparently Healthy

Infertility is the inability of a couple to achieve pregnancy over an average period of one year despite adequate, regular and unprotected sexual intercourse. Avitaminosis E has been implicated in the development of infertility and hemolytic anemia in animals. There is, however, little evidence that man is ever scanty in vitamin E. The aim of this study is to evaluate serum α-tocopherol levels and anemia among infertile patients attending Specialists Hospital, Sokoto. A total of fifty (50) infertile patients and fifty (50) apparently healthy fertile married as control were recruited for this study. The blood samples collected were analyzed for α-tocopherol using Hashim and Schuttringer (1996) method, and PCV using hematocrit reader. The data obtained were analyzed using the Student's T-independence test. The result shows that the level of serum α-tocopherol was significantly lower (p<0.05) in infertile patients (0.65±0.04) compared to controls (1.38±0.02). The result, however, shows no statistically significant difference of PCV and BMI (37.58±0.4 and 21.14±0.34 respectively) in infertile patients when compared to the controls (38.61±0.4 and 22.05±2.64 respectively). A reduced serum α-tocopherol level among infertile patients was observed in this study. The α-tocopherol has been described to be a potent antisterility factor on account of the development of the sterility in its absence. We, therefore, commend the incorporation of α-tocopherol in the diagnosis and treatment of infertility in human.

scholarly journals Assessment of chicken peripheral blood mononuclear cells isolated from freshly drawn blood versus 24 h refrigerated blood

2021 ◽  
pp. 2549-2553
Author(s):  
Chananphat Tantikositruj ◽  
Anchalee Buadkhunthod ◽  
Jatuporn Rattanasrisomporn ◽  
Warangkana Kitpipit ◽  
Chaiwat Boonkaewwan
Keyword(s):  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Gradient Centrifugation ◽  
Blue Dye ◽  
Fresh Blood ◽  
Peripheral Blood Mononuclear ◽  
Blood Samples ◽  
Significant Difference ◽  
Over Time

Background and Aim: The peripheral blood mononuclear cell (PBMC) is an excellent cell source for in vitro studies, particularly those involving immunology. The aim of this study was to determine the quality and quantity of chicken PBMCs isolated from freshly drawn blood as well as blood that had been chilled for 24 h. In addition, the survival of PBMCs cultured in medium was investigated. Materials and Methods: Blood samples were collected from 12 Betong and 12 Leghorn chickens. Hemograms were analyzed. Density gradient centrifugation was used to isolate PBMCs. PBMCs (2×106 cells/mL) were cultured in a culture medium and incubated in a CO2 incubator for 5 consecutive days. The number of viable cells was determined using the trypan blue dye exclusion method. Results: Blood samples were obtained from healthy chickens. There was no statistically significant difference in the total amount of PBMC between fresh and refrigerated blood samples from both chicken breeds. The viability of PBMCs isolated from fresh blood (95%) was significantly greater than blood refrigerated for 24 h (90-92%) in both breeds. Furthermore, the viability of PBMCs isolated from both blood samples decreased significantly over time, from 90-95% to 60-65%. Conclusion: The total number of PBMC in fresh and refrigerated blood was not significantly different. Fresh blood-derived PBMCs had significantly higher viability than 24 h refrigerated blood PBMCs. Furthermore, the viability of PBMCs decreased significantly over time.

scholarly journals Assessment of the Spine in a Healthy Working Population : A Radiographic Study of the Lumbrosacral Angle in Relation to Occupation in Southern Nigeria

2013 ◽  
Vol 5 (2) ◽  
pp. 99-105
Author(s):  
Michael Omon Oyakhire ◽  
Chukwu Agi
Keyword(s):  
Undergraduate Students ◽  
Statistical Significance ◽  
Medical Science ◽  
Control Group ◽  
Job Experience ◽  
Healthy Working ◽  
Significant Difference ◽  
One Year ◽  
Working Adult ◽  
Age Range

Objective: This study was carried out to evaluate the effect of occupation on the Lumbosacral angle of the spine in a cohort of working adult Nigerians Method: Lateral radiographs from two hundred and twenty healthy working adults within the age range 18 to 55 years comprising 136 males (61.82%) and 83 females (38.18%), with a mean age=29.14(7.45) years were studied. Participants at the time of investigation were engaged in different physically demanding occupations with a minimum of one year job experience. The sample included a control group made up of undergraduate students from the University of Port Harcourt. Prior to data collection, all participants read and signed informed consent forms. Each lateral radiograph was evaluated using Fergusons sacral base-horizontal 2-line method. Statistical analysis was done using the computer based SPSS Version 17, Chicago IL, and a confidence level of 0.05 was accepted as indicative of statistical significance. Results: A statistically significant difference was found between the four groups in terms of age, gender and occupation. The lumbosacral angle of the groups engaged in physically demanding occupations were significantly higher than the control (undergraduate students). Conclusion: The lumbar curves are designed to increase the resilience of the vertebral column. Over a time period, physically demanding jobs are likely to cause various degrees of significant alterations in the lumbosacral configuration DOI: http://dx.doi.org/10.3126/ajms.v5i2.8888 Asian Journal of Medical Science, Volume-5(2) 2014: 99-105

Risk of venous thromboembolism following surgical treatment of superficial venous incompetence

VASA ◽  
2017 ◽  
Vol 46 (6) ◽  
pp. 484-489 ◽  
Author(s):  
Tom Barker ◽  
Felicity Evison ◽  
Ruth Benson ◽  
Alok Tiwari
Keyword(s):  
Venous Thromboembolism ◽  
Varicose Vein ◽  
Lower Incidence ◽  
Varicose Veins ◽  
Secondary Data ◽  
Foam Sclerotherapy ◽  
Increased Risk ◽  
Significant Difference ◽  
Chi Squared ◽  
One Year

Abstract. Background: The invasive management of varicose veins has a known risk of post-operative deep venous thrombosis and subsequent pulmonary embolism. The aim of this study was to evaluate absolute and relative risk of venous thromboembolism (VTE) following commonly used varicose vein procedures. Patients and methods: A retrospective analysis of secondary data using Hospital Episode Statistics database was performed for all varicose vein procedures performed between 2003 and 2013 and all readmissions for VTE in the same patients within 30 days, 90 days, and one year. Comparison of the incidence of VTEs between procedures was performed using a Pearson’s Chi-squared test. Results: In total, 261,169 varicose vein procedures were performed during the period studied. There were 686 VTEs recorded at 30 days (0.26 % incidence), 884 at 90 days (0.34 % incidence), and 1,246 at one year (0.48 % incidence). The VTE incidence for different procedures was between 0.15–0.35 % at 30 days, 0.26–0.50 % at 90 days, and 0.46–0.58 % at one year. At 30 days there was a significantly lower incidence of VTEs for foam sclerotherapy compared to other procedures (p = 0.01). There was no difference in VTE incidence between procedures at 90 days (p = 0.13) or one year (p = 0.16). Conclusions: Patients undergoing varicose vein procedures have a small but appreciable increased risk of VTE compared to the general population, with the effect persisting at one year. Foam sclerotherapy had a lower incidence of VTE compared to other procedures at 30 days, but this effect did not persist at 90 days or at one year. There was no other significant difference in the incidence of VTE between open, endovenous, and foam sclerotherapy treatments.

Markers of Hemostatic Activation in Affected Coronary Arteries during Angioplasty

1994 ◽  
Vol 72 (05) ◽  
pp. 672-675 ◽  
Author(s):  
Nicolas W Shammas ◽  
Michael J Cunningham ◽  
Richard M Pomearntz ◽  
Charles W Francis
Keyword(s):  
Coronary Artery Disease ◽  
Coronary Artery ◽  
Venous Blood ◽  
Balloon Inflation ◽  
Blood Samples ◽  
Hemostatic System ◽  
Significant Difference ◽  
Hemostatic Markers ◽  
Artery Disease ◽  
High Doses

SummaryTo characterize the extent of early activation of the hemostatic system following angioplasty, we obtained blood samples from the involved coronary artery of 11 stable angina patients during the procedure and measured sensitive markers of thrombin formation (fibrino-peptide A, prothrombin fragment 1.2, and soluble fibrin) and of platelet activation ((3-thromboglobulin). Levels of hemostatic markers in venous blood obtained from 14 young individuals with low pretest probability for coronary artery disease were not significantly different from levels in venous blood or intracoronary samples obtained prior to angioplasty. Also, there was no translesional (proximal and distal to the lesion) gradient in any of the hemostatic markers before or after angioplasty in samples obtained between 18 and 21 min from the onset of the first balloon inflation. Furthermore, no significant difference was noted between angioplasty and postangioplasty intracoronary concentrations. We conclude that intracoronary hemostatic activation does not occur in the majority of patients during and immediately following coronary angioplasty when high doses of heparin and aspirin are administered.

Effect of Concentration of Trisodium Citrate Anticoagulant on Calculation of the International Normalised Ratio and the International Sensitivity Index of Thromboplastin

1994 ◽  
Vol 72 (01) ◽  
pp. 084-088 ◽  
Author(s):  
E M Duncan ◽  
C R Casey ◽  
B M Duncan ◽  
J V Lloyd
Keyword(s):  
Reference Material ◽  
Oral Anticoagulants ◽  
Reference Method ◽  
Trisodium Citrate ◽  
Sensitivity Index ◽  
Blood Samples ◽  
Orthogonal Regression ◽  
Significant Difference ◽  
International Normalised Ratio ◽  
International Sensitivity Index

SummaryThe aim of this study was to determine whether the concentration of trisodium citrate used to anticoagulate blood has an effect on the INR of the sample and the ISI of the thromboplastin. Five thromboplastins including and Australian reference material were used to measure the prothrombin time of normal and patient samples collected into two concentrations of trisodium citrate - 109 mM and 129 mM. There was no effect of citrate concentration on the INRs determined with the reference material. However for the other four thromboplastins there was a significant difference between INRs for the two citrate groups. The prothrombin times of the samples collected into 129 mM were longer than those collected into 109 mM. This difference was only slight in normal plasma but more marked in patients receiving oral anticoagulants, causing the INRs for patient plasmas collected into 129 mM citrate to be higher then the corresponding samples collected into 109 mM citrate.From orthogonal regression of log prothrombin times by the reference method against each thromboplastin, we found that the ISI for each thromboplastin was approximately 10% lower when determined with samples collected into 129 mM citrate than with samples collected into 109 mM. These results suggest that the concentration of trisodium citrate used for collection of blood samples can affect the calculation of the INR and the calibration of the ISI of thromboplastin. This was found both for commercial thromboplastins prepared by tissue extraction and for a recombinant tissue factor.

Levels of Prothrombin Fragment F1+2 in Patients with Hyperhomocysteinemia and a History of Venous Thromboembolism

1997 ◽  
Vol 78 (05) ◽  
pp. 1327-1331 ◽  
Author(s):  
Paul A Kyrle ◽  
Andreas Stümpflen ◽  
Mirko Hirschl ◽  
Christine Bialonczyk ◽  
Kurt Herkner ◽  
...  
Keyword(s):  
Venous Thromboembolism ◽  
Thrombin Generation ◽  
Oral Anticoagulant Therapy ◽  
Oral Anticoagulants ◽  
Control Group ◽  
Prothrombin Fragment ◽  
Hemostatic System ◽  
Significant Difference ◽  
System Activation ◽  
One Year

SummaryIncreased thrombin generation occurs in many individuals with inherited defects in the antithrombin or protein C anticoagulant pathways and is also seen in patients with thrombosis without a defined clotting abnormality. Hyperhomocysteinemia (H-HC) is an important risk factor of venous thromboembolism (VTE). We prospectively followed 48 patients with H-HC (median age 62 years, range 26-83; 18 males) and 183 patients (median age 50 years, range 18-85; 83 males) without H-HC for a period of up to one year. Prothrombin fragment Fl+2 (Fl+2) was determined in the patient’s plasma as a measure of thrombin generation during and at several time points after discontinuation of secondary thromboprophylaxis with oral anticoagulants. While on anticoagulants, patients with H-HC had significantly higher Fl+2 levels than patients without H-HC (mean 0.52 ± 0.49 nmol/1, median 0.4, range 0.2-2.8, versus 0.36 ± 0.2 nmol/1, median 0.3, range 0.1-2.1; p = 0.02). Three weeks and 3,6,9 and 12 months after discontinuation of oral anticoagulants, up to 20% of the patients with H-HC and 5 to 6% without H-HC had higher Fl+2 levels than a corresponding age- and sex-matched control group. 16% of the patients with H-HC and 4% of the patients without H-HC had either Fl+2 levels above the upper limit of normal controls at least at 2 occasions or (an) elevated Fl+2 level(s) followed by recurrent VTE. No statistical significant difference in the Fl+2 levels was seen between patients with and without H-HC. We conclude that a permanent hemostatic system activation is detectable in a proportion of patients with H-HC after discontinuation of oral anticoagulant therapy following VTE. Furthermore, secondary thromboprophylaxis with conventional doses of oral anticoagulants may not be sufficient to suppress hemostatic system activation in patients with H-HC.

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