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Aim: The study was undertaken to compare the alterations in the hematological parameters in buffaloes suffering from Amphistomosis with normal buffaloes and to correlate it with the subclinical infection that is hard to diagnose. Materials and Methods: Blood samples from 50 amphistomes infected as well as 50 non-infected buffaloes from slaughter houses were taken into vacutainer tubes containing ethylene diamine tetraacetic acid for estimation of various hematological parameters by Automatic Analyzer Hema-2062 manufactured by Analytical Technologies Ltd. Result: There was a significant reduction in the mean hemoglobin, total leukocyte count, total erythrocyte count and packed cell volume and significant increase in the neutrophils count and eosinophil count of infected buffaloes as compared to the non-infected buffaloes respectively. Conclusion: Amphistomosis is characterized by severe neutrophilia, eosinophilia, and anemia. Anemia of high intensity along with hepatic damage can lead to the death of the animal in severe cases. Alterations in the Hematological parameters can be used as an indicator to diagnose and check the severity of amphistomosis especially in young ones and in subclinical infection.

Background: Due to the high prevalence of hemolysis in specimens received from the emergency department (ED), several strategies have been proposed to improve sample quality, but none of these seem effective to overcome the problem. In a preliminary study we showed that the use of S-Monovette blood collection system was effective to lower the risk of hemolysis in venous blood samples collected from intravenous catheters. This study was hence aimed to verify whether the replacement of a conventional vacuum system with S-Monovette may be effective to reduce the bur den of hemolysis in the daily practice of a large urban ED. Materials and methods: The study was divided in two observational periods of 4 months each. In the former period, blood was collected from intravenous catheters using BD Vacutainer SST II Plus plastic serum tubes, whereas in the latter period the blood was drawn from intravenous catheters using S-Monovette blood tubes in aspiration mode. Sample hemolysis was automatically assessed in all serum samples by photometrical measurement. Results: The total number of hemolysed serum specimens was 624/14155 (4.41%) in the first phase of the study, and 342/13319 (2.57%) in the se cond phase of the study (P < 0.001). Conclusion: Results of our study confirm that the introduction of the Sarstedt S-Monovette blood tubes has reduced the hemolysis rate in the emergency department compared to the previously used BD Vacutainer® SST II Plus plastic serum tubes.

Incorrect blood sample volume-anticoagulant ratio has been the cause of both haematological and immunological errors especially when K3 EDTA-containing blood collection tubes were used. Lower whole blood sample volumes collected into 4.0 millilitres spray-dried K2 EDTA has been shown to overcome incorrect haematology results when analysed on automated haematology analyzers but there is no experimental evidence for the same in the CD4 count enumeration by flow cytometric technique. 9.0 ml of whole blood was collected from each of fifteen retroviral and ten normal volunteers and aliquot into five different 4.0 ml plastic spray-driedK2 EDTA blood collection tubes containing 4.0, 2.0, 1.5, 1.0 and 0.5 ml respectively. Each well-mixed sample was analysed on Partec Cyflow counter within 4 hours of collection for absolute CD4+T lymphocyte count.Results: Both the reference sample volume 4.0 ml and experimental lower sample volumes (2.0,1.5, 1.0 and 0.5 mls) of retroviral volunteers in 4.0 ml plastic spray-driedK2 EDTA blood collection tubes gave comparable CD4 count results with percentage mean difference of 1.82%, -1.48%,2.25% and 0% for 2.0 ml, 1.5 ml 1.0 ml and 0.5 ml respectively. Irrespective of sample volumes, the normal volunteers had higher CD4 count results. There was no statistically and clinically significant difference in the CD4 counts and the percentage mean difference were 0.4%, 0.17%, 1.00% and 0.23% for 2.0 ml, 1.5 ml, 1.0 ml and 0.5 ml respectively. The correlation (slope)and modest logistic regression coefficient (R2) of experimental lower sample volumes of both retroviral and normal volunteers were between 0.9500 and 1.0000 showing excellent agreement in the CD4 counts of both reference and experimental sample volumes(p

Introduction. Chronic peritoneal dialysis (PD) up-regulates vascular endothelial growth factor (VEGF) synthesis and VEGF is found in drained dialysate (dd). Objective. Aims of this prospective study were to evaluate serum (s) and ddVEGF concentration during the first six months of PD, relationships between these concentrations and demographic and biochemical parameters, presence of diabetes, peritonitis, and the use of medications. Methods. The study included 20 patients, with the mean age of 62.9?12.69, 11 of whom were affected by diabetes mellitus. Fasting venous blood samples were taken at the beginning and after six months of PD, in tri-potassium ethylenediaminetetraacetic acid (K3EDTA) vacutainer. Results. After six months of PD, sVEGF concentrations increased significantly, without significant change in ddVEGF. Concentrations of sVEGF at the beginning of chronic PD treatment directly significantly correlated with serum fibrinogen, and after six months with fibrinogen and glycemia. In patients receiving erythropoiesis-stimulating agent (ESA), levels of sVEGF and ddVEGF were lower at baseline, while after six months of PD ddVEGF increased. In patients not receiving ESA, sVEGF increased more prominently, while ddVEGF decreased. The changes were not statistically significant. Patients receiving angiotensinconverting- enzyme inhibitor (ACEi) had sVEGF and ddVEGF levels insignificantly lower than those not using ACEi, however sVEGF significantly increased during six months of PD. After six months of PD, ddVEGF was significantly higher compared to those not using ACEi. Treatment with statins did not significantly influence levels of sVEGF and ddVEGF during the follow-up. Concentrations of sVEGF were continually lower than those of ddVEGF and increased more, while concentrations of ddVEGF were higher in patients using statins. Conclusion. Serum and drained dialysate concentrations of VEGF in PD patients were connected with poorer metabolic profile, while the role of inflammation and treatment agents should be studied further.

Background Selection and verification of blood collection tubes is an important preanalytical issue in clinical laboratories. Today, gel tubes are commonly used with many advantages, although they are known to cause interference in immunoassay methods. In this study, we aimed to compare SSTs of two different suppliers (Ayset clot activator & Gel and Becton Dickinson (BD) Vacutainer SST II advance) with reference tubes and evaluate the effect of storage time in terms of commonly used endocrine tests such as thyroid-stimulating hormone (TSH), free thyroxine (fT4), and free triiodothyronine (fT3). Methods Fifty-five volunteers were included in the study. Samples were taken into three different tubes and analyzed for serum TSH, fT4, and fT3 on Architect ci8200 Immunoassay System. Clinical decision levels were estimated using total allowable error (TEa). Results No difference was found between tubes in terms of TSH, fT3, and fT4 levels. From a statistical standpoint, TSH and fT4 levels were no longer stable during 24, 48, and 72 hours storage time periods. However, their variations were not clinically significant. Conclusion Ayset clot activator & Gel tubes and BD Vacutainer SST II advance tubes have comparable results with glass tube in terms of TSH, fT3, and fT4 levels on Architect ci8200 Immunoassay Systems. From a clinical standpoint, serum TSH, fT4, and fT3 concentrations may be considered as stable when storing these tubes over 72 hours.

Background Storage of blood samples after centrifugation , decapping and initial sampling allows ordering of additional blood tests. The pre-analytic stability of biochemistry and immunochemistry analytes has been studied in detail, but little is known about the post-analytical stability in incurred samples. Methods We examined the stability of 23 routine analytes on the Dimension Vista® (Siemens Healthineers, Denmark): 42–60 routine samples in lithium-heparin gel tubes (Vacutainer, BD, USA) were centrifuged at 3000 × g for 10 min. Immediately after centrifugation, initial concentration of analytes were measured in duplicate (t = 0). The tubes were stored decapped at room temperature and re-analyzed after 2, 4, 6, 8 and 10 h in singletons. The concentration from reanalysis were normalized to initial concentration (t = 0). Internal acceptance criteria for bias and total error were used to determine stability of each analyte. Additionally, evaporation from the decapped blood collection tubes and the residual platelet count in the plasma after centrifugation were quantified. Results and conclusion We report a post-analytical stability of most routine analytes of ≥ 8 h and do therefore – with few exceptions – suggest a standard 8 hour-time limit for reordering and reanalysis of analytes in incurred samples.

Peripheral blood mononuclear cell (PBMC) isolation is commonly done via density gradient centrifugation over Ficoll-Hypaque, a labor-intensive procedure that requires skilled technicians and can contribute to sample variability. Cellular Preparation Tubes (CPTs) are Vacutainer blood draw tubes that contain Ficoll-Hypaque and a gel plug that separates the Ficoll solution from the blood to be drawn. Once blood is drawn into CPTs, they can be centrifuged to separate the PBMC, then shipped (if desired) to a processing lab. The processing lab removes the PBMC from the upper compartment of the tube (above the gel plug), washes the PBMC, and can cryopreserve them using DMSO-containing media, as detailed in this protocol.

Background: Serologic testing for antibodies against epitopes from pathogens is a valuable tool for investigating the relationship between infection and disease. This study comprehensively evaluates the impact of preanalytic variation on antibody seropositivities to a selected set of antigens arising from delays in processing of blood samples, preprocessing storage temperature, and vacutainer type. Methods: We assessed peripheral blood collected from 29 volunteers in four different Vacutainer types [ethylenediaminoetetraacetic acid (EDTA), acid-citrate-dextrose (ACD), lithium heparin (LH), serum separator tubes (SST)], and stored at 4°C or room temperature for 0, 1, 2, 3, 4, 5, and 6 days before processing. Multiplex serology was used to determine antibody reactivity against 35 antigens derived from human papillomaviruses, human polyomaviruses, Epstein–Barr virus, and Helicobacter pylori. Cohen's κ statistic was used to measure agreement on seropositivity status between samples exposed to standard and nonstandard clinical practice conditions. Results: For samples processed without delay, κ was not associated with storage-temperature (P value range 0.23 to 0.95) or vacutainer type (P value range, 0.35–0.89). Kappa did not significantly decline with increasing delays in processing for any vacutainer-type storage temperature combination (P slope range, 0.06–1.00). Conclusions: Antibodies to epitopes from various pathogenic infectious agents can be measured reliably from samples stored in SST, EDTA, ACD, or LH vacutainers at either room temperature or 4°C for up to 6 days before processing. Impact: Serologic testing is robust to several preanalytic options. These findings are particularly important for epidemiologic studies recruiting participants from remote settings where sample exposure to preanalytic conditions can vary considerably. Cancer Epidemiol Biomarkers Prev; 26(8); 1337–44. ©2017 AACR.

Recently, because the quality of laboratory analyses has increased along with the need for quality improvement, several external quality control bodies have adapted performance specifications using the Desirable Biological Variation Database, termed “Ricos goals”; these criteria are more stringent than those presented in CLIA 88. In this study, we aimed to validate newly introduced serum separator tubes, Improvacutor, for routine clinical chemistry testing in accordance with Ricos goals and CLIA 88. Blood samples were collected from 100 volunteers into three types of serum vacuum tubes: Greiner Vacuette, Becton Dickinson (BD) Vacutainer, and Improve Improvacutor. The samples were subjected to 16 routine chemistry tests using a TBA-200fr NEO chemistry autoanalyzer. In the comparison analysis, all 16 test results were acceptable according to CLIA 88. However, in the comparison of Improve and BD tubes, creatinine showed 4.31% (+0.08 μmol/L) bias. This slightly exceeded the Desirable Specification for Inaccuracy Ricos limit of ±3.96%, but still satisfied the CLIS88 limit of ±26.52 μmol/L. The remaining 15 analytes performed acceptably according to the Desirable Specifications of Ricos. The correlation coefficient of 12 analytes was greater than 0.95 in Passing-Bablok regression analysis among the three tubes, but was lower for four analytes: calcium, sodium, potassium, and chloride. In the stability assay, only potassium tested in the Greiner tube revealed a larger positive bias (2.18%) than the Ricos Desirable Specification for Inaccuracy based on biologic variation (1.8%). The BD tube also showed a positive bias of 1.74%, whereas the new Improve tube showed the smallest positive bias of 1.17% in potassium level after 72 h storage. Thus, the results of this study demonstrate that recently introduced analytical performance specifications based on components of biological variation (Rico's goal) could be extended to criterion for performance evaluation and applied.

Introduction Although serum-providing blood tubes with a barrier are still widely used due to their significant advantages, the use of blood tubes with a barrier to provide plasma is becoming widespread. We compared 22 analytes in a BD Vacutainer® Barricor LH Plasma tube for local clinical validation of this new lithium heparin tube with a barrier. Materials and methods Samples from 44 volunteers were collected in different tubes (Becton Dickinson and Company): Z tube without additive (reference), clot-activator tube with gel (SST), lithium heparin tube without gel (LiH), and lithium heparin tube with barrier (Barricor). Analyte concentrations in different tubes were compared with the reference tube. All tubes were also evaluated according to additional testing (different centrifugation durations, blood-sampling techniques and individual differences). Results Aspartate aminotransferase (AST), glucose (Glc), potassium (K), lactate dehydrogenase (LD), sodium (Na), and total protein (TP) had a significant bias in Barricor (9.19%, - 3.24%, - 4.88%, 21.60%, - 0.40%, 5.03%, respectively) relative to the reference tube. There was no statistical difference between different centrifugation durations and individual differences for AST, K and LD in LiH and/or Barricor (P > 0.05). There was a significant bias for LD between LiH and Barricor in terms of blood-sampling techniques (21.2% and 12.4%, respectively). Conclusions Recently, the use of plasma has become prominent due to some of its advantages. In this study, plasma AST, K, LD, Glc and TP levels in Barricor were clinically different in comparison to serum. The results of additional tests showed that higher levels of LD in Barricor did not result from haemolysis, and they might be related to other factors including number of platelets, cellular fragility, or functional environment.

Background Intact parathyroid hormone (PTH) tests are frequently sandwich immunoassays. Enzymes that cleave PTH may cause falsely lower PTH results. The objective of this study was to determine whether bovine thrombin in Becton Dickinson (BD) Vacutainer rapid serum tubes™ (RSTs) may lead to PTH results that are lower than in plasma separator tube™ (PST) or serum separator tube™ (SST) collections. Methods Tubes of blood (PST, SST, and RST) were collected from donors. PTH concentrations were measured on a Roche Cobas e602 analyzer in aliquots held at room temperature or 4 °C across time. Instrument comparison studies were also conducted on an Abbott Architect i1000SR and a Siemens Immulite 2000 XPi. Previously collected serum specimens were also incubated in exogenous bovine thrombin, the direct thrombin inhibitor hirudin, or both. Freshly collected RST specimens were also spiked with hirudin after clotting and centrifugation. Results Significant decreases in PTH degradation rate constants were observed according to tube type, with degradation rates faster in RSTs than SSTs, and SSTs faster than PSTs. PTH degradation rate was temperature dependent. PTH decreases induced by exogenous bovine thrombin, as well as endogenous human thrombin, were reduced by hirudin. Conclusions Bovine thrombin is responsible for the decrease in PTH results observed in RSTs. Endogenous human thrombin, activated during clot formation, is likely responsible for the smaller decreases observed in non-RST sera versus plasma.

AimsTo investigate the suitability of newly developed Becton Dickinson Vacutainer rapid serum tube (RST) for therapeutic drug monitoring of antibiotics and anticonvulsants.MethodsTwo pools of citrated whole blood were created by spiking high and low concentrations of gentamicin, vancomycin, phenytoin, lamotrigine and carbamazepine. After recalcification with 15 mmol/L calcium chloride, spiked whole blood was added into four different Becton Dickinson blood collection tubes: RST, serum separator tube, red top tube and polyethylene plain tube. Serum aliquots were collected at baseline (0 h), 2 h, 24 h, day 3 and day 7. Drug concentrations were measured in batch by HPLC and the Architect c8000.ResultsGentamicin and vancomycin concentrations were stable up to 7 days in all 4 blood collection tubes. Anticonvulsants results for the RST were stable and did not deviate substantially from those of the red top and plain tubes, and demonstrated better performance than the serum separator tubes that showed significant (≥10% bias, pConclusionsThe RST provides acceptable drug stability over the course of 7 days for gentamicin, vancomycin, phenytoin and lamotrigine and over 3 days for carbamazepine.

Summary Reasons for performing study No recommendations have been made regarding the relative timing of blood collection for autologous conditioned serum (ACS) preparation and surgical procedures. Objectives 1) To identify effects of surgical stress on cytokine levels in ACS, 2) identify haematological markers for prediction of cytokine production in ACS and 3) investigate the necessity for specialised ACS containers when preparing a cytokine-rich serum. Study design Experimental in vitro study. Methods Blood was drawn from 15 stallions admitted for elective castration preoperatively and 22–24 h post operatively and incubated in ACS containers and plastic vacutainer tubes containing Z Serum Clot Activator. Concentrations of interleukin (IL)-1 receptor agonist (IL-1Ra), IL-10, IL-1β, tumour necrosis factor (TNF)-α, insulin-like growth factor (IGF)-1 and transforming growth factor (TGF)-β were determined in all serum samples and compared between preparation methods and sampling time by ANOVA. Changes in cytokine levels induced by incubation, defined as delta cytokine, were calculated by subtracting the baseline levels from the levels in incubated samples. Based on post operative serum amyloid A (SAA), horses were grouped into ‘mild‘, moderate’ and ‘marked’ surgical stress; delta cytokine levels in post operative samples were compared between these groups by ANOVA. Results Delta IGF-1 was significantly lower in post operative samples compared with preoperative. Horses in the ‘marked’ surgical stress group had significantly lower delta IL-1Ra and delta TGF-β than the ‘moderate’ group and significantly lower delta IGF-1 than the ‘mild’ group. No association between cytokine levels and haematology variables were identified. Cytokine levels were comparable between serum prepared in blood tubes and in specialised ACS containers. Conclusions Surgical stress influences the cytokine content in ACS. Useful predictors of cytokine production in ACS were not identified. Specialised ACS containers may not be necessary for preparation of a cytokine-rich serum.

Background: Obtaining accurate, precise, and reliable platelet counts as a reference for the calibration of hematology analyzers has been a continuing problem due to varying results. We proposed the relativity of platelet count by Automated Cell Counter and Manual Haemocytometer with Peripheral smear for focusing the accuracy and & cost effectiveness, without compromising the accuracy of test results. Methods: Seventy-Five blood samples in EDTA-anticoagulant vacutainer tubes were obtained and randomly divided into 3 group; Group I-normal control (n = 25), Group II-mild thrombocytopenia (n = 25) and Group III-severe thrombocytopenia (n = 25). Each blood sample had two peripheral blood smears made and stained on an automatic slide stainer. Peripheral blood smears were analyzed for platelet count by manual microscopy, an automated hematology analyzer Beckman Counter Diff 2 in all groups. The agreement between the two methodologies was assessed using the paired t-test and correlation coefficient analyses. Results: The coefficient of variation values significantly differed in Group C and was high for automated cell counter. High positive correlation was also observed between Automated cell counter values and manual method (r=0.82, p

Background: Auricular therapy is a promising treatment for pain. However, the physiologic mechanisms of analgesic effects are not well-understood, which limits the scientific credibility of auricular therapy for pain management. Objectives: This prospective, randomized clinical trial (RCT) was conducted to determine whether or not the levels of pro- and anti-inflammatory cytokines change in response to auricular point acupressure (APA) for chronic low-back pain (CLBP). Methods: Blood samples (10 mL) were collected in a vacutainer, based on standard phlebotomy procedures. Blood was drawn at the following timepoints: before APA treatment to measure the baseline; 30 minutes after the first APA treatment; weekly for 4 weeks; and within 1 month of a follow-up visit (a total of seven timepoints) for each subject). Results: Participants with CLBP reported a mean 70% reduction of pain intensity at the completion of the 4-week APA regimen. The participants also had changes in serum pro- and anti-inflammat...

Introduction: Standard guidelines assert that any vacutainer, which is 10% less under-filled should be rejected as it may affect the results. These guidelines refer to the studies using liquid K 3 -ethylene diamine tetra acetic acid (EDTA). Although the new guidelines recommend K 2 -EDTA instead of earlier K 3 -EDTA but has not clearly commented upon whether the same holds true for the new anticoagulant K 2 -EDTA. Materials and Methods: Blood samples from 100 healthy blood donors were included. We took 7-8 ml of blood and was then transferred to three 3 ml capacity vacutainers-up to 3 ml, 2.5 ml, 1 ml (marked a, b, c respectively) containing spray-dried K 2 -EDTA as anti-coagulant. Thus, b and c vacutainers were under-filled. Hematological parameters in all the samples were analyzed immediately using automatic analyzer (Cell Dyn). Statistical analysis was done with the help of SPSS v. 17.0 (Chicago, IL, USA) using one-way ANOVA. Results: The results showed that three groups were comparable with respects to hemoglobin, hematocrit, total leukocyte count, differential leukocyte counts, total platelets, mean corpuscular volume, mean hemoglobin concentration, mean cellular hemoglobin concentration along with red cell distribution width. Conclusion: Our study suggested that under-filling till 67% (1 ml in 3 ml standard capacity) of the recommended volume in a spray-dried K 2 -EDTA vacutainers did not affect hematological parameters in healthy people. In view of these results, there is need to further investigate the same in diseased. If further studies done on under filling of the pathological samples also give similar results as in our study then only the present guidelines related to under filling could be revised when K 2 -EDTA is used as an anticoagulant.

This study was carried out to determine the comparative study of cd8+ cell count and leukopoietin levels in human immunodeficiency virus infection in Umuahia, Nigeria. A total number of ninety (90) subjects within the age of 18-60 years were used for the study. The subjects were divided into three (3) groups of thirty (30) subjects in each group. Group I comprised of control subjects. Group II were HIV- subjects not on therapy, and group III were AIDS- subjects not on therapy. Blood samples were collected into commercially prepared dipotassium EDTA vacutainer for both test and control subjects after informed consent. These were used in determining their CD8+cells counts and Leukopoietin levels. After the analyses of the blood samples, the following results were observed. The CD8+cells count of group II (967 +198 cells/ml) was significantly higher than that of group I (586+81 cells/ml) while the CD8+cells count of group III CD8+ (1086+228 cells/ml) showed a statistically higher level when compared with those of groups I & II at p

Aims: To determine the association between dyslipidemia and MGD. Settings and Design: It is a prospective, observational study of 1 year duration. Thirty patients who were diagnosed with meibomian gland dysfunction and 30 age and sex matched controls were taken into study after they gave their informed consent and taking into consideration the inclusion and exclusion criteria. Methods and Material: After enrollment of the subject to the study all subjects had undergone a thorough ophthalmic examination. Tear film break up time, Schirmer's Test without anesthesia followed by with anesthesia was calculated. Statistical analysis used: Meibomian gland status was evaluated by, expressibility, and numerical scoring of staining. test was done by taking venous sample of blood in a red-topped vacutainer after overnight fasting. Values of total cholesterol (TC), high-density lipoprotein (HDL) , low-density lipoprotein (LDL), and triglycerides (TGs) were noted. Results: There exists an association between increasing stage of MGD and age and increasing values of TC,LDL,TGs of lipid profile.Conclusions: Higher stages of MGD are associated with increased age. MGD and increasing levels of LDL, total cholesterol, and triglycerides have a positive correlation.

The objective of the current study was to determine the effect of pre-slaughter stress, season and breed on the activity of plasma creatine kinase (CK) and the quality of mutton. One hundred and seventy-three (173) castrated sheep from Dormer (DM), South African Mutton Merino (SAMM), Dorper (DP) and Blackhead Persian (BP) sheep breeds were used in the study. The animals were grouped according to age-groups as follows: Group 1 (6 to 8 months), Group 2 (9 to 12 months) and Group 3 (13 to 16 months). Blood samples were collected during exsanguinations using disposable vacutainer tubes for CK analysis. Representative samples of the Muscularis longissimuss thoracis et. lumborum (LTL) were collected from 84 castrated sheep, of different breeds (28 per breed) 24 h after slaughter. The following physico-chemical characteristics of mutton were determined; meat pH (pH24), color (L*, a* and b*), thawing and cooking losses and Warner Braztler Shear Force (WBSF). The activity of plasma CK was significantly higher (p0.001) levels (1,358.6±191.08) of CK. South African Mutton Merino had higher values for pH24 (5.9±0.06), L* (34.2±0.97), b* (12.2±0.50) and WBSF (26.8±1.51) and Blackhead Persian had higher values (35.5±2.17) for cooking loss (CL%) than the other breeds. Computed Principal Component Analyses (PCA) on the activity of plasma CK and physico-chemical characteristics of mutton revealed no correlations between these variables. However, positive correlations were observed between pH24, L*, a*, b*, CL% and WBSF. Relationships between pre-slaughter stress, CK activity and physico-chemical characteristics of mutton were also observed. It was therefore concluded that although mutton quality and creatine kinase were not related, pre-slaughter stress, season and breed affected the activity of creatine kinase and mutton quality.

Background Current sampling and transport conditions of samples in general practice can result in pseudohyperkalaemia. This study was undertaken to determine, in a general practice setting, whether there is any difference in haemolysis obtained when using BD Vacutainer® Rapid Serum Tubes (BD RST) compared with using BD Vacutainer® SST™ II Advance Blood Collection Tubes (BD SSTII). Methods Blood was collected from 353 patients requiring blood sampling who were attending 31 general practitioner practices in Belgium. For each patient, two BD SSTII tubes and two BD RST tubes were drawn in a randomized order. One of each pair of tubes was inverted five times, the other was not. Serum potassium concentration, serum LDH activity and haemolysis index were measured in each sample. Results There was no significant difference in measured potassium concentration according to tube type ( P = 0.16). Measured LDH activities were 1.7% higher in serum collected into BD SSTII tubes compared to BD RST tubes ( P = 0.02). When comparing serum from unmixed BD RST with BD SSTII tubes, there was a slight reduction in the haemolysis index but no significant difference in measured potassium concentration or LDH activity. Risk of hyperkalaemia was 4.8 times higher in serum from tubes that were incompletely filled compared to those that were filled with the correct amount of blood. Conclusion Both types of blood tubes are suitable for the measurement of serum potassium and LDH in patients from general practice. Tube inversion does not improve the accuracy of either serum potassium or LDH measurement. Blood tubes should be filled to the level recommended by the manufacturer to avoid artefactual increases in measured serum potassium concentration and LDH activity.

Contents The study aimed to compare the acid–base balance and steroid concentrations between follicular fluids (FF) of pre-ovulatory follicles derived from a spontaneous oestrus (SO), synchronized or induced oestrus (IO) and follicular cysts (CYS) and between FF and blood in dairy cows. Forty-two dairy cows were included in this study. The animals were allocated to three groups: SO (n = 23); IO (n = 11) using GnRH at day 0 and day 9 and PGF2α at day 7; and animals with CYS (n = 10). The follicular fluids (FF) were aspirated from the cyst/pre-ovulatory follicles (∅ ≥ 15 mm) after SO and after second GnRH dose in IO by transvaginal ultrasound-guided ovum pick-up technique. Blood samples (BL) were collected in heparinized vacutainer tubes. The oxygen tension (pO2) in FF of IO was higher (p

Introduction: A fatty acid (FA) is a carboxylic acid with a long aliphatic chain, which is either saturated or unsaturated. Recently, the role of FA and particularly omega-3 and -6 has emerged as cardiovascular risk factor in the literature. The aim of our study was to establish reference values for these FA and to compare them with data obtained in a population of acute myocardial infarction (AMI) patients. Materials and methods: Hundred thirty five healthy subjects (59.38 ± 27.12 yo, 75 men) were selected as reference population. We also evaluated FA in thirty three patients (55 ± 9 yo, 23 men) admitted in the Emergency Department of our Institution for AMI. The fasting whole blood was drawn in vacutainer containing EDTA. Before analysis, samples were washed and transmethylated. We performed the quantification of different FA by gas chromatography associated with flame ionization detector (GCFID). Results: We obtained results in control healthy patients to be used as reference values. In the AMI group, levels of omega-6 were significantly higher (p 0.05) for C18:2n6 and C18:3n6 than the reference population and omega-3 values were significantly lower (p 0.01) compared to reference value for C22:6n3. The omega-3 index was lower and the ratio omega-6/omega-3 was higher in AMI group compared to reference values. Conclusions: We have established reference value for FA and have compared these values with the results obtained in AMI population. FA determination is a new tool we are able to use and to process in our laboratory which can help the clinician to screen patients with the highest cardiovascular risks because of the implication of FA in the etiopathogeny of atherosclerosis.

Objectives To investigate potential interferences of two pre-analytical variables, the storage time and the vacutainer tube, on the quantification of 20 amino acids using a UHPLC-MS/MS method. Design and methods Blood samples from 25 apparently healthy subjects were collected into duplicate sets of EDTA-2K, EDTA-3K, coagulation, heparin and citrate tubes, and stored in capped vacutainer tubes at 4 °C for 6 h, 12 h and 24 h before sample analysis. A UHPLC-MS/MS method was established for simultaneous quantification of 20 amino acids. ANOVA for repeated measurement was conducted based on the model of Mauchly's test of Sphericity. Student's t-test was applied for comparison between amino acid concentrations obtained from different vacutainer tubes, and consistency of the results was checked through Bland-Altman difference plots and Passing-Bablok regression analysis. Results Most of the 20 amino acids showed a least concentration fluctuation with storage time in heparin plasma, followed by EDTA-3K and citrate plasma. The amino acid concentrations were significantly lower in citrate plasma and slightly higher in serum, compared with those in heparin plasma. No fixed bias was observed for amino acid concentrations in EDTA and heparin plasma, but the differences were mostly of statistical significance. Amino acid concentrations in EDTA-3K plasma achieved a good consistency with those in heparin plasma by UHPLC-MS/MS analysis. Conclusions Storage time and vacutainer tube were important variables for amino acid analysis. They should draw researchers' attention and then be controlled in good laboratory practice to reduce pre-analytical errors.

BackgroundVery limited information is available about the prevalence of blood groups among Qatari population and residents. The information about frequencies of different blood group antigens is important to manage, patients who has tendency to develop alloantibodies such as thalassemics. Thalassemia is most common genetically inherited blood disorder due to a quantitative defect in hemoglobin formation, results ineffective erythropoiesis, leads to severe anemia requires regular blood transfusions. To maintain a safe blood supply for alloimmunized patients are difficult, without the knowledge of donor pool's antigen frequency evidence. We have determined the frequencies of the Rh, and Kell, clinically significant antigens among blood donors in Qatar.MethodAs per approved IRB and consent waiver signed, blood samples were collected from voluntary blood donors at Hammad Medical Cooperation (HMC), Blood Centre in EDTA vacutainer tubes. The samples were, then processed on day of collection to remove plasma and the packed RBC (pRBC) was 3 times washed with saline. After washing the pRBC were used to make 5% cell suspension with saline. Blood samples were collected from regular healthy 512 donors. Blood group antigens were analyzed by tube and Gel card method as manufacturer's instruction. The Rh antigens including, D, C, E, c, e and Kell (K) were typed and incidence frequencies were expressed as percentage. Anti-sera used in tube testing were bought from Lorne Laboratories, United Kingdom (UK) and the Gel cards from Grifols® The results were analyzed by using statistical software SPSS. The data were analyzed using descriptive exploratory features to see various blood group antigens we have studied and characterizedResultsWe analyzed blood group antigens; with age range from 19–65 years, mean donors age 38.4 ± 8 years. Nationalities among donors varied considerably, the highest frequency donations were Qataris 89 (17.4%), Syrians 89 (17.4%) and Egyptians 77 (15%). When compared among Arabs 365 (72.1%) and non-Arabs 132 (25.8%), majority of blood donors are from Arab countries. Rh antigens frequency shown from our analysis is, D 88.08%, C 70.11%, E 33.59%, c 75.39%, e88.47% and K 10.4%. The Rh antigens among Qataris were D 92.13% E 39.32%, e 88.76%, C 65.16%, c 86.5%, K 13.8%.ConclusionsOur data shows the blood donors in state of Qatar are multinational and demonstrates variations in frequency of Rh and Kell RBC antigens. These variations may be a challenge to find antigen negative blood, needs for multiple transfused alloimmunized patients such as thalassemics and sickle cell patients. It is important to establish the incidences of various RBC antigens amid blood donors, when dealing with patients who have developed multiple antibodies. The current clinical practice in blood banks is randomly cross match the available blood units in the stock. This study will help to provide data regarding the frequency of Rh and Kell blood group clinically significant antigens. As we have mentioned very limited information are available, about blood group prevalence among Qatari population accept some data about the association of blood groups among diabetes mellitus patients in Qatar. Our study shows though there is large number of expatriates in Qatar, among them Arabs are the highest frequent blood donors including Qataris. While Indians resident community is the largest, among expatriates here in state of Qatar number of blood donation is less compare to other residents. Our data shows Qataris and Syrians 17.4% are the most regular blood donors in Qatar. The age groups highest frequent blood donors were 25–34 years in Qatar, but according to WHO facts blood donation, among high income countries most regular blood donation is in the age group 45–64 years, while low and middle income countries the age group donates most regularly are 18–25 years.

For the subsequent analysis of the methylated mSEPT9 colorectal cancer screening marker in plasma, different blood collection tubes and blood storage conditions were investigated. The study demonstrated that methylated Septin 9 (mSEPT9) can be consistently detected in plasma samples derived from whole blood samples collected with S-Monovette® K3E and BD Vacutainer ® K2EDTA tubes stored at 2-8 °C for a maximum of 24 h and for samples collected in S-Monovette CPDA tubes stored at 18-25 °C for up to 48 h.

Uric acid has stronger pro-oxidant than antioxidant properties during equine endurance events and thus, limits performance and has serious repercussions on health. The aim of the study was to investigate the changes in uric acid, leucocytes, plasma biochemical parameters and heart rate in metabolic endurance horses. Thirty Arabian endurance horses were physically examined and blood samples were collected pre and post-race. After physical examination, the successfully completed (n = 10) and metabolic disordered ( n = 20) endurance horses were identified. Blood samples in heparinised vacutainer tubes were used for the determination of uric acid, triglyceride, creatine kinase, aspartate transaminase, packed cell volume, lactate, total protein and plasma protein. Blood sample in ethyl diaminotetra-acetic acid vacutainer tubes were used for the analysis of leucocytes. The age, body weight, heart rate, humidity and ambient temperature were also recorded. One way Analysis of variance and pairwise correlations were used for the analysis. A value of P ≤ 0.05 was considered as significantly different. The mean values of uric acid, lactate, leucocytes, plasma protein, total protein, heart rate, creatine pinase and Packed cell volume were significantly different between the successfully completed and metabolic dis - ordered endurance horses P < 0.0001), respectively. The mean values of aspartate transaminase and triglyceride were significantly different between the successfully completed and metabolic disordered endurance horses: P < 0.0130 and P < 0.0004, respectively. There were significant positive correlations between uric acid and lactate (r = 0.5196; P < 0.0271), between uric acid and plasma protein (r = 0.6025; P < 0.0175), between uric and Packed cell volume (r = 0.5206; P < 0.0268), between uric acid and triglyceride (r = 0.5541; P < 0.0170) and between uric acid and heart rate (r = 0.5629; P < 0.0150) in the metabolic disordered endurance horses. In conclusion, heart rate, triglyceride, blood lactate and packed cell volume were significantly associated with uric acid, a biomarker of oxidative stress. Therefore, uric acid could be used to evaluate performance and health status in endurance horses during training and endurance events.

Summary Reasons for performing study: The stability of total CO2 concentration (ctCO2) in plasma is influenced by storage temperature and handling during sample processing. Conflicting information exists regarding the stability of ctCO2 in equine plasma over time, and the effect of centrifugation on the measured value for plasma ctCO2 is unclear. Objectives: To determine plasma ctCO2 stability over 5 days when equine blood is collected into Vacutainer tubes, centrifuged within 30 min of collection, and stored at 4°C; and to determine whether a delay in centrifugation increases the rate at which plasma ctCO2 decreases over time. Methods: Blood was collected from 6 adult horses into 3 ml plastic Vacutainer tubes and randomly assigned to be centrifuged immediately, or after, storage. Plasma ctCO2 was measured in triplicate at 0, 24, 48, 72 and 96 h after collection using a NOVA-4 analyser. Data were analysed using multivariable linear regression, with P

International normalized ratio (INR) discrepancies were noted between clinical laboratories using various prothrombin time (PT) systems. We studied the influence of different commercial blood collection tubes and different PT systems on INR measurements. INRs of fresh patient samples were determined by 3 laboratories, each using different PT systems. In the first part of the study, samples were drawn with Vacutainer tubes and in the second part with Monovette tubes. In the first part of the study, the maximum bias for all patients amounted to 0.46 INR (14%), and in the second part, to 0.14 INR (4.9%). The maximum bias for all patients could be reduced further by local system calibration using frozen pooled plasma specimens. The sodium citrate solutions in the blood collection tubes were contaminated with magnesium ions (approximately 2.7 mmol/L and 0.3 mmol/L in the Vacutainer and Monovette, respectively). INR discrepancies could be explained largely by this influence of blood collection tubes. The maximum allowable magnesium contamination in sodium citrate anticoagulant solutions should be less than 1 mmol/L.

Vacutainer CPT tubes require blood samples for TSPOT.TB to be processed within 8 h. In this study we evaluated the ability of T-Cell Xtend to maintain the number and function of lymphocytes after 24 and 48 h of blood storage, giving similar test results as in freshly isolated specimens.Whole blood specimens from 59 individuals were collected in Vacutainer CPT tubes (CPT) and lithium heparin (LH) tubes. CPT tubes were processed within 8 h. T-Cell Xtend was added to LH tubes after 24 or 48 h. We also left LH tubes untreated for 48 h. Total number of white blood cells (WBC) and proportions of lymphocytes and granulocytes were determined in the isolated Peripheral Blood Mononuclear Cells (PBMC). We also evaluated the performance of T-Cell Xtend in the TSPOT.TB assay.PBMC yields from T-Cell Xtend treated LH samples did not differ from PBMC yields from CPT tubes, but T-Cell Xtend had a pronounced effect on the proportions of lymphocytes and granulocytes. The mean lymphocyte percentage in PBMCs isolated from fresh CPT blood was 84.31 ± 1.14% (at t = 48 h), but was decreased to 52.72 ± 3.34% (p0.05) in untreated LH blood (at t = 48 h). This effect was neutralized by T-Cell Xtend (85.44 ± 0.74%). We observed a similar but opposite effect on granulocytes: The mean proportion in untreated LH blood was increased to 40.9 ± 3.67% (p0.001) compared to CPT blood (8.26 ± 0.89%). Treatment of LH samples with T-Cell Xtend (48 h) restored the proportion of granulocytes to 8.47 ± 0.61%. Enumeration of spots in the TSPOT.TB assay demonstrated good agreement between CPT and T-Cell Xtend results, even after 48 h.T-Cell Xtend efficiently removes granulocytes from PBMC suspensions and increases the proportion of lymphocytes. TSPOT.TB results from T-Cell Xtend treated blood samples are at least comparable to the results obtained from the current CPT method. Use of standard lithium heparin blood combined with T-Cell Xtend allows up to 48 h storage of blood samples for batched processing and may further decrease the rate of indeterminate TSPOT.TB results.

Context.—Clinical samples that have densities greater than that of separator gel in specimen tubes may exhibit gel flotation to the top of the specimen upon centrifugation. Inappropriate separator gel flotation can occur in specimens with high protein content. In automated analytical systems, gel flotation can lead to mechanical disruption and potential inaccurate result reporting upon aspiration into instrument sampling probes. Objective.—To determine the relative specimen densities and estimated total protein contents at which specimen gel flotation would occur upon centrifugation in commonly used commercial specimen tubes, a comparative study of separator gel density was initiated using prepared dextran solutions. Design.—Specific gravity of several dextran solutions was determined by direct hydrometry. The dextran solutions were introduced to serum and plasma lithium heparin BD Vacutainer specimen tubes manufactured by Becton, Dickinson and Company and into Vacuette specimen tubes manufactured by Greiner Bio-One containing separator gel. Following centrifugation the specimen tubes were examined for gel flotation. Results.—Flotation was observed at a lower dextran solution density for Greiner than for BD tubes in both serum and plasma separator gel specimen tubes. Additionally, some differences between specimen tube lots were observed for both BD and Greiner tubes. The total protein content in clinical samples that would result in gel flotation can be estimated for different specimen container types. Conclusions.—Differences were observed for the gel separator specific gravity in different blood collection containers. Laboratories wishing to avoid problems with inappropriate gel flotation in high protein samples should consider these observations.

Summary Reasons for performing study: Osteoarthritis (OA) is one of the most prevalent and debilitating conditions affecting the horse. Autologous conditioned serum (ACS), commercially available as IRAP and IRAP II, is a recently developed treatment for OA in which plasma is prepared from venous blood by incubation with glass beads for 24 h. This product has been shown to increase anti-inflammatory cytokines and growth factors in human blood. However, data for equine ACS preparations are lacking. Objectives: To characterise the protein profiles produced by commercially available ACS systems in equine blood. Methods: Blood was drawn from 5 horses into 6 groups: red top vacutainer (control), IRAP and IRAP II, with and without heparin. Samples were collected 1 or 24 h post draw and analysed for IL-1ra, IL-10, IGF-1, TGF-β, TNF-α and IL-1β using ELISAs. Results: Twenty-four hour IRAP and IRAP II samples contained significantly higher levels of all cytokines relative to 1 h serum controls. At 24 h, IRAP II contained significantly higher levels of IL-1ra and IRAP contained significantly higher levels of TNF-α, compared to 24 h controls. In addition, TGF-β, IL-10 and IL-1β in IRAP and IRAP II sera were similar to 24 h serum controls. The addition of heparin significantly reduced levels of IGF-1, TNF-α and TGF-β, and significantly elevated levels of IL-1ra. Conclusions: The cytokine profile that IRAP II produced is modestly better than IRAP. Incubation of whole blood in glass tubes stimulated cytokine synthesis, although not as efficiently as IRAP II. Potential relevance: Although high levels of IL-1ra were found in ACS, elevation of other factors suggests these cytokines play a previously understated role in clinical improvements. Because ACS has been shown to alleviate clinical symptoms of OA, the present study suggests that factors other than IL-1ra alone might be involved in its clinical efficacy. Species-dependent elevations of cytokines warrant further investigation and optimisation of the systems appears to be necessary based on the differences between human and equine blood.

Introduction: Serum blood collection tubes with separator gel are widely used by many laboratories for chemistry analyses. We describe a case of a primary blood collection tube filled with blood sample and a floating separator gel. Materials and methods: The blood sample was collected from a 51 years old female in intensive care unit with the diagnosis of pneumonia into a BD Vacutainer SST tube (Becton Dickinson, NJ, USA) containing serum separator gel and conveyed to the core laboratory of Marmara University Hospital within 30 minutes from collection. Sample was immediately centrifuged at room temperature at 1500 × g for 10 minutes. Results: The analyses revealed a highly increased total protein concentration of 145 g/L (reference interval 64–83 g/L). The nephelometric analyses showed an elevated serum IgG concentration of 108 g/L (reference interval 6.5–16 g/L) and IgG lambda monoclonal band was determined by serum immunofixation electrophoresis. Conclusion: Limitation of the separator gel tubes in patients with a high plasma density and its possible effects on test results and laboratory costs should be remembered. The clinical diagnosis stated in the information system should also reveal known comorbid conditions besides the apparent admission reason. This information would avoid resampling, additional testing, and communication efforts with the clinicians.

The purpose of this study was to evaluate the influence of exercise on plasma tryptophan (TRP) and free serotonin (f5-HT), whole blood-5-HT (WB-5-HT) and f5-HT/WB-5-HT ratio in Italian Saddle horses. Six clinically healthy Italian Saddle horses were subjected to a 450 meters obstacles course. Blood samples were collected from each horse by jugular venipuncture using vacutainer tubes with K3-EDTA at rest, immediately after exercise, and after 30 min. TRP, f5-HT and WB-5-HT were analyzed by HPLC. Immediately after exercise, statistically significant increases of f5-HT (p

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is now widely used for marker/multibiomarker detection in medical diagnosis. We tested a new protocol for bacterial identification from blood culture broths in hospital routine by using collection tubes with separator gels on 503 included samples examined over 3 months, where 1.5 mL was injected by a syringe into BD Vacutainer tubes from BACTEC-positive bottles, before processing for bacterial protein extraction. Samples were loaded in duplicate onto the MALDI MS target, allowing a series of 12 samples to be processed in duplicate within 80 min by using Biflex III and BioTyper 2.0 software (Bruker). Including polymicrobial samples, 193 of 213 of Gram-negative bacteria (91.08%) and 284 of 319 of Gram-positive bacteria (89.02%) were correctly identified at the species level. Enterobacteriaceae constituted 35.15% of all species found, Staphylococaceae 37.96%, Streptococaceae and Enterococaceae 20.85%, Pseudomonadaceae 1.69%, and anaerobes 2.44%. In most of the polymicrobial samples, one of the species present was identified (80.9%). Seven isolates remained misidentified as Streptococcus pneumoniae, all belonging to Streptococcus mitis. Staphylococcus aureus was identified better when grown on anaero-aerobic medium, and MALDI BioTyper identification scores as low as 1.4 were pertinent, provided that four successive proposals of the same species were given. This new protocol correlates with conventional microbiology procedures by up to 90%, and by >95% for only monomicrobial samples, and provides a decreased turn-around time for identification of bacteria isolated from blood cultures, making this technology suitable also for blood cultures, with less delay and cost decreases in bacterial diagnostics, and favouring better care of patients.

Summary Reasons for performing study: Plastic heparinised Vacutainer® (registered trademark of Becton, Dickinson and Company) tubes are used for blood gas analysis in horses. This collection method may not be ideal because influx of atmospheric O2 through the permeable plastic wall of the Vacutainer® tube and loss of CO2 into the gas phase above the blood sample should increase blood PO2 and decrease PCO2, respectively. Objectives: To determine the effects of collecting blood into plastic Vacutainer® tubes and storage conditions on blood gas analysis values. Methods: Blood was obtained from 6 healthy horses and tonometered at 37°C with 12% O2 and 5% CO2. Three ml aliquots of tonometered blood were collected using a glass syringe or Vacutainer® tube and stored in iced water or at room temperature for 0, 5, 15, 30, 60 and 120 min. Blood samples from Vacutainer® tubes were collected aerobically (tube opened for 5 s) or anaerobically (tube remained closed). Blood gas analysis was performed in duplicate using a Radiometer ABL5. Data was analysed using repeated measures analysis of variance and P

Aim Analysis of circulating donor derived cell-free DNA (ccfDNA) promises a non-invasive method of assessing allograft rejection and possibly eliminating the need for surgical biopsies. In this study we compared the concentration and purity of ccfDNA in specimens collected in ACD (BD Vacutainer) tubes and Cell-Free DNA BCT (Streck) tubes and processed after different storage times. Methods Blood samples from 4 healthy donors were drawn into collection tubes (3 tubes each for analysis at different time points). Samples were stored at room temperature and processed at 18, 24, and 72 h. The samples were processed using Promega LV ccfDNA Custom kit according to the manufacturer protocol. Tubes were centrifuged at 2000 g for 10 min and an average of 4 ml of plasma was separated from the cells. Samples were extracted on the Maxwell RSC, and eluted in a volume of 65 ul. Samples were measured by Qubit (Thermo Fisher) & ProNex DNA QC Assay (Promega) to determine DNA concentration. The level of genomic DNA (gDNA) contamination was determined by the ProNex Assay and confirmed by the TapeStation D1000 High Sensitivity kit (Agilent Technologies). The ProNex Assay uses a multiplexed probe-based qPCR assay to detect quantity and quality of genomic DNA. It detects 75 bp, 150 bp and 300 bp genomic DNA sequences. The presence of 300 bp sequences indicates contamination with gDNA. Results DNA concentration by the Qubit showed a wide range of quantity. The quantity of 75 bp and 150 bp DNA fragments as shown by the qPCR assay indicate the amount of ccfDNA in each sample. The ratios of smaller fragments to 300 bp in each sample indicate the level of gDNA contamination. The Streck specimens had consistently high ratios up to 72 h. The ACD specimens had much lower ratios, decreasing by 72 h. The presence of gDNA contamination was confirmed by TapeStation traces and one such example at 72 h is shown below. Conclusions ACD tubes consistently exhibited higher gDNA contamination, as reflected by lower ratios in qPCR, which increased over time. Download : Download high-res image (190KB) Download : Download full-size image

OBJECTIVE: This study investigated the quality of trauma specimens by comparing line draws to venipuncture. DESIGN: The draw type (line or venipuncture); container type (Vacutainer or Microtainer); and suitability for processing (processed/hemolyzed/clotted) of routinely collected trauma specimens was analyzed. SETTING: The clinical laboratory of a Level I Pediatric Trauma Center. MAIN OUTCOME MEASURE: Hemolyzed trauma specimens were analyzed according to method of collection, collector, and type of container to identify issues resulting in unusable samples. RESULTS: The data shows that for 13% of all draws, portions of the results were affected by hemolysis. Sixteen percent of line draws and 6% of venipunctures were hemolyzed (p = 0.04). There was no statistical association with who collected the sample (p = 0.07) or type of container (p = 1.00). CONCLUSION: Based on this sample of data, the laboratory recommends that, whenever possible, venipunctures be performed for laboratory testing of blood specimens to improve trauma specimen integrity. ABBREVIATIONS:ED = Emergency Department; EMS = Emergency Medical Services; ID = identification; MLA = medical laboratory assistant; RBC = red blood cells; RN = registered nurse.

The aim of this study was to evaluate the stability of blood concentrations of a variety of illegal and medicinal drugs that are important for forensic analyses when spiked and stored in Vacutainer or Venosafe evacuated plastic collection tubes compared to Vacutainer evacuated glass tubes. Tubes were filled with spiked whole blood and analyzed after storage for one week at ambient temperature and at -20 degrees C, respectively. Freeze-and-thaw stability was included in the study. No significant difference between storage in glass or plastic tubes was noted for any compound investigated.

BACKGROUND Biomarker assays are often conducted on whole blood samples in the course of drug development studies. Because bacterial lipopolysaccharide (LPS) (endotoxin) contamination is known to cause spontaneous cytokine production by monocytes, contamination of blood collection tubes may interfere with biomarker assay results. METHODS Whole blood from healthy donors was collected into plastic or glass sodium (Na+)-heparin Vacutainer™ blood collection tubes and heparinized syringes. Samples were analyzed for phosphoprotein response, cytokine production, and RNA expression. Tubes were tested for endotoxin contamination by use of the limulus amoebocyte lysate assay. RESULTS Results of phospho-flow cytometry, branched DNA (bDNA), and ELISA assays indicated that a specific lot (#5339582) of plastic Na+-heparin Vacutainer tubes was highly contaminated with an endotoxinlike substance, and contamination was confirmed by the limulus amoebocyte lysate assay. Analysis of multiple-analyte panels revealed that analytes whose changed expression was predictive of LPS stimulation were increased when whole blood was incubated in contaminated tubes for 6 or 18 h. Two additional lots of plastic tubes tested had detectable amounts of endotoxin sufficient to strongly alter phospho-flow cytometry analyses, as determined by the fold change in phosphorylation of p38 mitogen-activated protein kinase in response to tumor necrosis factor α and LPS. In contrast, 3 lots of glass tubes had substantially lower levels of spontaneous blood activation. CONCLUSIONS Endotoxin contamination associated with tubes from 3 lots of a particular type of plastic Na+-heparin Vacutainer tube dramatically affected biomarker assay measurements. Prescreening these tubes is suggested before their use in clinical sample analysis.

Peripheral blood film (PBF) is an important laboratory workup for clinical characterization of haematological malignancies that involves blood smear on microscope slide and cytology of blood cells. PBF preparation, typically manually operated, requires technical training and empirical experience and is not without human errors. Automated blood smearing by traditional spin coating has been an alternative. However, with intended use originally for semiconductor industries, it utilizes volatile organic solvents, employs powerful centrifugation at high speed, which frequently alters morphology and damages cells, and inherently creates radius dependent pattern. In addition, manual blood dispensing could bring contamination and spin-off of excess blood as aerosol can be hazardous. While improvement of traditional spin coating technology to overcome intrinsic defects and compensate operational variations is possible, low-speed spin coating is more predicable and controllable for uniform PBF thickness. An innovative automatic PBF coating system has been developed to smear monolayer of dispersed blood cells for cytopathology and thick PBF of multilayered blood cells as well for identification, enumeration, and examination of rare tumor cells in peripheral blood. Explicitly, whole blood is dispensed through nozzle directly from the blood collection tube that slides radially on rails above substrate, and coated on the spinning substrate at low speed in non-contact mode circle-by-circle with no intra/inter- circular overlaps and negligible centrifugal force. Consequently, there is no spin-off, saving blood samples and preventing potential biohazard. External contamination is also eliminated by dispensing blood directly from the tube. Temperature control of substrate is further enabled to preserve cellular viability and morphology. At speed of 60 rpm, contact angle of 10°, stepper of 0.5 mm, and room temperature, monolayer PBF with intact cellular morphology across the entire substrate was created using healthy fresh whole blood directly from the BD Vacutainer K2-EDTA tube. To study tumor cell heterogeneity in circulation, thick PBFs have been evenly coated on 100 mm diameter discs using 2 mL of pre-stained, cancer patient blood samples at speed of 30 rpm, contact angle of 45°, stepper of 1 mm and elevated temperature of 37°C and fully dried. PBFs were then scanned by our innovative Spinning Disc Imaging system (iSDI) to pinpoint fluorescent nucleated cells of epithelial origin. Images of suspects were automatically displayed with identities of location, area, orientation, width, height, and signal intensity. Results demonstrated tumor cells with variable sizes, shapes and expression levels of EpCAM, in addition to clusters and fragments. Despite advances in genetic and molecular diagnostics, PBF remains an indispensable part of diagnostic jigsaw in unraveling heterogeneity and evolution of tumor in liquid phase. Citation Format: Andy Cheng, Leo Xu, Bin Hong. Spin coated peripheral blood film reveals tumor heterogeneity in circulation [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 211.

Objective: To assess the impact of cannula valve connectors on haemolysis of blood samples drawn from newly inserted cannulae. Methods: In a semi-blinded, randomized study paired blood samples, with and without cannula valve connector, were obtained from patients within the ED and tested for haemolysis, defined as haemolysis index of greater than 120 mg/dL. Patients were randomized as to which sample was collected first. Cannula size was standardized and vacutainer systems provided consistent draw pressures. Time taken for the tube to fill was recorded as a measure of blood flow. Results: Two hundred and ninety patients were randomized, with six subsequently excluded from analysis because of samples being lost or insufficient for testing. Average patient age was 60.8 years and 52.5% were male. There were no significant differences between the randomization groups. The overall rate of haemolysis was 2.6%, being 2.8% in the valve first group and 2.5% in the no valve first group (P= 1.0). Time for collection averaged 7.7 s in the valve first group and 7.5 s in the no-valve first group (P= 0.22). Mean serum potassium level was 4.4 mmol/L in both groups (P= 0.46). The rate of hyperkalaemia was not different between valve first and no-valve first groups (12.7% and 13.7%, respectively, P= 1.0). Conclusion: The attachment of a cannula connector valve to a peripheral cannula prior to blood sampling is not associated with an increase in the rate of haemolysis or hyperkalaemia.

Problem statement: Smoking plays a significant role in the developmen t of periodontal disease. Quantitative relation between smoking and increased severity of periodontal disease, by means of biochemical marker has not been described in Malaysian population. The present study was designed to apply serum cotinine measurement as a quantitative method to evaluate smoking levels in Malaysian patients and to correlate these levels with the severity of periodontal disease. Approach: The study group consisted of 80 healthy individual s (20-64) year, Current Smokers 26, Non Smokers 27 and Former Smokers 27. The subjects were then asked to complete a questionnaire including the demographic, socioeconomic status, medical history and history of cigarette smoking. The periodontal variables recorded were amount of Visible Plaque score, gingival bleeding Index and community periodontal index. Samples of blood "10 mL" were obtained in vacutainer tubes containing EDTA for quantitative analysis of serum levels of cotinine. The serum samples were analyzed for cotinine content by means of a competi tive-inhibition ELISA technique. Results: Current smokers represent the highest mean cotinine serum l evel, 95.5 ng mL -1 , compared to former smokers, 35.5 ng mL -1 and non smokers, 22.9 ng mL -1 . The mean serum cotinine level in periodontally healthy patient showed the highest co tinine level (84 ng mL -1 ) followed by the gingivitis patients (68 ng mL -1 ) and (50 ng mL -1 ) for periodontitis patients. Conclusion: The present observations clearly indicate an association betwee n smoking, periodontal disease clinical parameters "plaque, gingival bleeding scores" and cotinine ser um levels in current smokers. Cotinine serum levels doesn't affected by the existence or the severity o f periodontal disease.

Progesterone (P4) is an important component of oestrous cyclicity and is critical to fertility. A concentration >1 ng mL−1 reflects the function of the corpus luteum (CL) and is considered indicative of a cyclic cow. Recent publications have shown that P4 at the onset of synchronization programs is critical to pregnancy outcomes in primiparous cows (Stevenson et al. 2015 J. Anim. Sci. 93, 2111-2123) and cows with P4

Serum separator tubes were introduced 35 years ago and were widely used in the clinical laboratory in China for routine collection of blood because of providing a closed system that allowed for collection, transport, processing, sampling, and storage of specimens. This type of tubes facilitated rapid separation of serum from cellular constituents of blood and also prevented hemolysis upon prolonged storage. However, there were some limitations associated with gel tubes (i.e., gel and analyte stability). In order to circumvent these problems, BD released a new serum separator tube containing a new gel (BD SST(™) II Plus). We investigated theperformance of BD SST(™) II Plus tubes for tumor marker tests using BD Serum Glass tubes as controls. Equivalence between the BD SST(™) II Plus and BD Serum Glass tubes was demonstrated for all analytes at initial time. Also, all analytes remained stable when stored in BD SST(™) II Plus tubes up to 72 hr. Concentration of neuron‐specific enolase tended to increase with preservation time up to 72 hr in BD Serum Glass tubes. We conclude that BD SST(™) II Plus was suitable for collection of blood and storage of serum for tumor marker tests. J. Clin. Lab. Anal. 24:418–421, 2010. © 2010 Wiley‐Liss, Inc.

Reactive carbonyl species (RCS) have been widely used as indicators of oxidative stress. However, the associations of carbonyl stress with aging process and biochemical alteration of erythrocyte are still poorly understood. Fresh blood samples in vacutainer tubes containing sodium heparinate were obtained from 874 volunteers who were divided into young, adult and old groups based on their age. Plasma RCS and thiols concentrations between different age groups and erythrocyte membrane protein carbonylation in the adult group were detected within 24 h of the blood sampling. Results showed that the plasma thiols concentration decreased gradually during aging process, and the p-values between all three groups are less than 0.05. The plasma RCS concentration in different age groups showed a nonlinear association with age. The levels in the young group were slightly higher than the adult group (not significant) and lower than the old group (p < 0.01). The protein carbonylation of erythrocyte membrane was positively correlated with plasma RCS concentration (p < 0.01), but not plasma thiols concentration. We conclude that higher levels of RCS, not lower levels of thiols, in plasma are a direct risk factor for the protein carbonylation of erythrocyte membrane. Owing to the decrease of thiols levels and increase of RCS levels during aging process, a shift from RCS-related redox allostasis to carbonyl stress would contribute to age-related biological dysfunction and even aging process.