Lithium Heparin Research Articles

Reference Intervals for Plasma Free Kappa and Lambda Chains and Kappa/Lambda Ratio using PENIA.

Determination of free light chains is used increasingly for patients with multiple myeloma and other plasma cell dyscrasias. The aim of the present study was to define reference intervals for free kappa and lambda chains and kappa/lambda ratio in lithium heparin plasma and compare the test results with cystatin C based estimated glomerular filtration rate. Free kappa and lambda chains were measured in Liheparin plasma from 222 healthy blood donors using free light chain reagents and a BNII nephelometer from Siemens Diagnostics. Calculated reference intervals for kappa chain, free was 4.73 mg/L (90% confidence interval 4.00 - 5.45) - 22.66 mg/L (20.33 - 24.98), for lambda chain, free was 4.33 mg/L (3.70 - 4.95) - 29.28 mg/L (26.96 - 31.59), and for Kappa/Lambda Chain ratio 0.59 (0.56 - 0.62) - 1.46 (1.37 - 1.56). There was no need for gender or age specific reference intervals. Cystatin C based estimated glomerular filtration rate had a significant effect on the levels of free light chains. The study presents reference intervals for plasma free light chains and compared results of estimated glomerular filtration rate with free light chains of kappa and lambda.

Using Residual Blood from the Arterial Blood Gas Test to Perform Therapeutic Drug Monitoring of Vancomycin: An Example of Good Clinical Practice Moving towards a Sustainable Intensive Care Unit.

Regarding sustainability in the intensive care unit (ICU), there is increasing interest in reducing material waste and avoiding unnecessary procedures. Therapeutic drug monitoring (TDM) of vancomycin, using a dedicated tube, is standard clinical care during treatment with vancomycin. Furthermore, in the ICU, on a daily basis, arterial blood gas (ABG) tests are frequently performed throughout the day. After analysis, a variable volume of blood is discarded. Lithium heparin (LiHep) syringes for ABG tests differ from normally used dipotassium ethylenediaminetetraacetic acid (K2EDTA) tubes. The primary objective was to compare both containers and validate the use of LiHep syringes. Secondary objectives were to evaluate the potential impact on saving materials, nursing time, and costs when implementing vancomycin TDM via LiHep syringes. Vancomycin analysis from sampling in lithium heparin (LiHep) syringes for ABG tests was validated and compared with the concentrations from conventional sampling in K2EDTA tubes. For method comparison, a Bland-Altman plot and Deming regression analysis were performed. The method was validated for inter- and intra-day precision and accuracy. Vancomycin was analyzed by means of the validated method using a particle-enhanced turbidimetric inhibition immunoassay (PETINIA) autoanalyzer. Furthermore, an analysis was conducted to evaluate the potential impact of implementing vancomycin sampling via ABG tests on savings in materials, nursing time, and costs. From 18 patients, 24 plasma samples in both K2EDTA tubes and LiHep syringes were obtained and compared. The mean relative difference between the two containers was -2.0% (-3.0 to -0.93%). Both the Deming regression analysis and the Bland-Altman plot met the acceptance criteria. Potentially, over 1000 blood draws and accompanying materials and packaging can be saved when vancomycin samples are obtained by means of scavenged LiHep syringes. The vancomycin analysis for LiHep syringes showed a total interday precision of 1.95% and an accuracy of 99.7%. The total intraday precision was 2.22%, and the accuracy was 99.2%. Accuracy and precision values were within the acceptance criteria of recovery 85 to 115% and ≤15%, respectively. No significant differences were found in vancomycin concentration between the two analyses, and the LiHep analysis was validated for further implementation in clinical care. Residual blood from ABG test samples can be used for TDM of vancomycin, resulting in a potential reduction of materials used and the number of blood draws. These results will contribute to a more sustainable TDM process with benefits for the patient.

Effects of time delay and blood storage methods on analysis of canine venous blood samples with an Element point-of-care analyzer.

Manufacturers of point-of-care (POC) analyzers recommend immediate processing and anaerobic collection of blood samples. However, it is not uncommon for clinical scenarios to result in delayed sample processing or room air exposure that could impact the test results. To investigate the effect of time delay and sample storage method on key POC analytes in canine venous blood samples processed with an Element POC analyzer. Blood gas analysis was performed on venous blood samples at times 0 (T0), 15, 30, and 60 minutes after sampling using three different storage methods: preheparinized plastic syringes and two different lithium heparin tubes. To determine clinical relevance, results were compared with allowable total error of the respective parameter. Significance was set at P < 0.05. Significant differences between the three storage methods at baseline were found for partial pressure of carbon dioxide (PCO2 ), partial pressure of oxygen (PO2 ), base excess, and total hemoglobin. No significant differences up to T60 were found within collection methods for actual bicarbonate (HCO3 - ), base excess, sodium, potassium, chloride, ionized calcium (iCa), glucose, and BUN. Significant differences within collection methods were found after T0 for creatinine, after 15 minutes for lactate, and after 30 minutes for pH and hematocrit. No significant differences were found for PO2 in samples stored in preheparinized plastic syringes at any time point. These results suggest that HCO3 - , sodium, potassium, chloride, iCa, glucose, and BUN are comparable within the three storage methods for up to 60 minutes after sampling without resulting in clinically relevant changes.

Impact of pre‐analytical factors on blood‐based biomarkers of Alzheimer's disease

AbstractBackgroundReliable blood‐based biomarkers (BBBMs) are urgently needed to allow for robust, cost‐effective pre‐screening of patients with suspected Alzheimer’s disease (AD) before referral for detailed assessments. We evaluated the effects of storage time and temperature, plastic tube types, and freeze/thaw (F/T) cycles in fresh, never frozen and previously frozen whole blood (WB) and plasma. Effects of anticoagulants and tube transfer were only studied in previously frozen plasma.MethodWB samples from patients with clinically suspected AD were collected in ethylenediaminetetraacetic acid (EDTA) tubes and stored at room temperature (RT) or 4°C for ≤24hrs as WB or plasma. Separated plasma was transferred into polypropylene (PP) tubes to evaluate the effects of tube transfer and F/T cycles, or into low‐density polyethylene (PE‐LD) tubes to assess the effect of tube type. In addition to EDTA, lithium heparin (Li‐Hep) and sodium citrate (NaCit) tubes were used and effects of anticoagulants evaluated. Plasma levels of phosphorylated‐tau (pTau) 217, pTau181, amyloid‐β 1–42 (Aβ42), amyloid‐β 1–40 (Aβ40), apolipoprotein E4 (ApoE4), glial fibrillary acidic protein (GFAP), and neurofilament light chain (NFL) were measured using automated Elecsys® prototype immunoassays (cobas e 601 analyzer; all Roche Diagnostics International Ltd).ResultFor all BBBMs, pre‐analytical effects were comparable between fresh, never frozen and previously frozen samples. All BBBMs were stable for ≤24hrs at 4°C in WB and EDTA plasma; Aβ42, Aβ40, and ApoE4 were unstable if stored for &gt;2hrs at RT. Up to five tube transfers, or two F/T cycles, and both tested plastic tube types were acceptable for all BBBMs. For all BBBMs, except Aβ42 and Aβ40, analyte levels were comparable between EDTA, Li‐Hep, and NaCit tubes. Aβ42 and Aβ40 were most sensitive to pre‐analytical sample handling and the effects could only be partially compensated by the Aβ42/Aβ40 ratio.ConclusionWe provide recommendations for an optimal handling protocol for blood collection and sample handling for analysis of BBBMs for AD based on measurement in fresh samples (Figure): WB and EDTA plasma should be stored at 4°C for ≤24hrs, while storage at RT should be avoided or limited to 2hrs maximum.

Whole blood and plasma taurine reference intervals in adult Cavalier King Charles Spaniels and correlations between taurine concentration, diet and mitral valve disease.

To determine breed-specific reference intervals for whole blood (WB) and plasma taurine concentrations in adult, overtly healthy Cavalier King Charles Spaniels (CKCSs) and determine whether taurine concentrations differ across preclinical myxomatous mitral valve disease (MMVD) stages or between CKCSs eating diets that meet World Small Animal Veterinary Association (WSAVA) nutritional guidelines versus other diets. 200 privately owned CKCSs. Clinically healthy adult CKCSs were recruited prospectively. Diet and supplement history was collected. Dogs were staged by echocardiography using MMVD consensus guidelines. Taurine concentrations were measured in deproteinized lithium heparin WB and plasma samples with the postcolumn ninhydrin derivatization method on a dedicated amino acid analyzer. There were 12 stage A (6%), 150 stage B1 (75%), and 38 stage B2 (19%) CKCSs. Seventy-eight dogs (39%) were reported by their owners to be eating diets meeting WSAVA nutritional guidelines; 116 (58%) were not. Taurine concentrations in plasma (P = .444) and WB (P = .073) were not significantly different across MMVD stages or between CKCSs eating diets meeting WSAVA nutritional guidelines versus other diets (P = .345 and P = .527, respectively). Reference intervals for WB taurine (152 to 373 µM) and plasma taurine (51 to 217 µM) concentrations in CKCSs were generated. In CKCSs, taurine concentrations do not differ significantly based on preclinical MMVD stage, nor do they differ significantly based on consumption of a diet that does or does not meet WSAVA nutritional guidelines.

Comprehensive Plasma Amino Acid Analysis Following Administration of Short- and Long-Acting Erwinia Asparaginase (Crisantaspase) Demonstrated Significant Increase in Plasma Serine and Glycine Levels

Background: Asparaginases are amidohydrolases that catalyze the conversion of asparagine (Asn) and glutamine (Gln) to aspartate (Asp) and glutamate (Glu), respectively, and can therefore be used therapeutically for cancer, particularly acute lymphoblastic (and myeloid) leukemia, to deplete circulating Asn and Gln from plasma. Clinically available asparaginases are derived from two bacterial sources: Escherichia coli and Erwinia chrysanthemi (also called crisantaspase), with crisantaspases demonstrating higher glutaminase activity. In order to identify any potential contributory pathways or resistance mechanisms to asparaginases, we sought to investigate the impact of short- and long-acting crisantaspases on circulating levels of plasma amino acids (AAs) other than Gln and Asn. Methods: Free AA concentrations were measured via a dedicated high-pressure liquid chromatography AAanalyzer (Biochrom 20 PlusAmino or Biochrom 30 Acid Analyzer). Free AAs were separated with a cation-exchange resin post precipitation of protein with sulfosalicylic acid. The AAs were differentially eluted after increasing the pH and ionic strength of the buffer. The concentration of each AA was measured by reaction of the AA with ninhydrin and detection via a colorimeter. Standard curves were generated for each AA and AAs were quantified and reported as μmol/L (µM) of AAs in plasma. For mouse studies, plasma was isolated from whole blood and delivered on ice to the Biochemical Genetics lab (University of Maryland Baltimore). For human clinical samples, 5-8 mL blood in a sodium or lithium heparin was obtained, kept on ice, and within 40 min of being collected, the plasma was separated by centrifugation (1100 g × 5 min at 4°C) and frozen at -20°C until analysis. Mouse plasma samples were from studies that included treatment with pegcrisantaspase (PegC), a long-acting Erwinia-derived asparaginase, in both immunodeficient and immunocompetent mice. Human plasma samples were from a clinical study (NCT02283190) in patients with acute myeloid leukemia (AML) treated with the short-acting crisantaspase, Erwinaze. Results: As expected, in our in vivo mouse experiments, intravenous pegcrisantaspase significantly decreased plasma Gln and Asn and increased plasma Glu. Interestingly, we observed that post-pegcrisantaspase administration, plasma serine (Ser) and glycine (Gly) increased in a dose-dependent manner. In immunocompromised mice treated with one dose of pegcrisantaspase (125, 250, and 500 IU/kg), three days post-administration, Ser increased from 190 ± 6 µM in the vehicle-treated group to 274 ± 35 µM (p= 0.07) in the 125 IU/kg group, 357 ± 29 µM (p=0.01) in the 250 IU/kg group, and 565 ± 8 µM (p=0.0004) in the 500 IU/kg group. Gly also significantly increased from 295 ± 25 µM in the vehicle-treated group to 438 ± 24 µM (p=0.02) in the 125 IU/kg group, 507 ± 50 µM (p=0.03) in the 250 IU/kg group, and 633 ± 8 µM (p=0.003) in the 500 IU/kg group (Fig 1). Pegcrisantaspase also increased plasma Ser from 143 ± 7.9 µM to 301 ± 73.7 µM (p=0.02) and Gly from 251 ± 45.9 µM to 381 ± 44.4 µM (p=0.02) in immunocompetent C57BL/6 mice. In addition to non-tumor/non-leukemia bearing immunocompetent and immunocompromised mice, this phenomenon was observed in a patient-derived xenograft model of AML (comprehensive plasma AA analysis, Fig 2). Other amino acids significantly increased by pegcrisantaspase in mice include threonine and α-amino-n-butyric acid. In a comprehensive plasma amino acid evaluation in samples from AML patients treated with Erwinaze (NCT02283190), we observed a similar pattern of increase in Ser, from 82 ± 11.2 µM at baseline to 128 ± 8.3 µM on Day 8 (p=0.0007), and Gly, from 191 ± 25.8 µM at baseline to 282 ± 46.6 µM on Day 8 (p=0.006), after 1 dose of Erwinaze (25,000 IU/m2). Systematic literature review of clinical studies with asparaginases revealed that only levels of Gln/Glu and Asn/Asp have been published post-asparaginase administration; demonstrating that this report is the first comprehensive analysis of plasma amino acid levels after asparaginase exposure. Conclusions: We report a comprehensive evaluation of plasma AA alterations in both mice and humans treated with crisantaspase. We observed a reproducible crisantaspase-induced increase in plasma Ser and Gly, which we are currently exploiting for synergism with other AML treatment modalities. Figure 1View largeDownload PPTFigure 1View largeDownload PPT Close modal

Alterations in Hepatocellular, Reproductive, and Oxidative Stress Parameters in Female Albino Rats Exposed to Crude Oil

Aim: To study the effect of exposure to crude oil on the liver, ovary, and some oxidative stress parameters in albino rats.&#x0D; Study Design: A total of 50 female albino rats were used in the experiment. The rats were grouped into three: The control group which consisted of 10 rats, the low dose group which consisted of 20 rats, and the high dose group also consisted of 20 rats. The low dosage group was orally administered 1.5 mL crude oil mixed with 300 grams of rat feeds (0.005 mL/g) and the high dosage group was orally administered 3.0 mL crude oil mixed with 300 grams of rat feeds (0.01mL/g), while the control group was fed with normal rat feeds. The treated feeds were given once a day for 35 days.&#x0D; Place of Study: The study was carried out in the Department of Medical Laboratory Science, Rivers State University, Port Harcourt, Nigeria.&#x0D; Methodology: On the 36th day, the rats were sacrificed and then 5mL of blood from each rat was collected by cardiac puncture into labeled lithium heparin bottles for liver enzymes assay, hormonal assay, and oxidative stress parameters assay, while the livers and ovaries were harvested and fixed in 10% formal saline before tissue processing and histological examinations using H&amp;E staining technique. The collected blood specimens were spun; the plasma was extracted and analyzed in the laboratory for Aspartate aminotransferase (AST), Alanine aminotransferase (ALT), Alkaline Phosphatase (ALP), follicle Stimulating hormone (FSH), luteinizing hormone (LH), Prolactin, Malondialdyde (MDA), and Superoxide dismutase (SOD). Statistical analysis was performed using Graphpad prism version 8.02.&#x0D; Results: Significantly higher plasma levels of AST, ALT, and MDA in the treated groups, except for ALP which was only significantly higher in the high-dose group. FSH, LH, Prolactin, and SOD indicated significantly lower levels in the crude oil-treated rats. The histological examinations showed marked distortion in the architecture of the livers and ovaries of the treated groups, also, there was a reduction in ovarian cellularity and massive degenerated tissues.&#x0D; Conclusion: It is shown that exposure to crude oil contaminants orally could have a significant effect on the plasma level of hepatocellular enzymes, reproductive hormones, and oxidative stress parameters which in turn could lead to hepatocellular dysfunction, infertility, or impaired reproduction in mammals and cellular injuries caused by excess free radicals as signaled by plasma level of oxidative stress parameters.

Correlation of Mean pH, HCO3 and CO2 Between Arterial Blood Gases and Venous Blood Gases in Critically Ill Patients

Objective: To determine the correlation of mean pH, HCO3 and CO2 between arterial and venous blood specimens in critically ill patients.&#x0D; Study Design: Cross-sectional study.&#x0D; Place and Duration of Study: Department of Chemical Pathology and Endocrinology, Armed Forces Institute of Pathology (AFIP), Rawalpindi, from Apr to Oct 2015.&#x0D; Methodology: Forty critically ill patients admitted to the Intensive Care Unit were selected. Analysis of arterial blood gases followed by venous blood gases of the same patient was carried out after collection in lithium heparin tubes. Specimens were analyzed on COBAS 221 fully automated ABGs and an electrolyte analyzer.&#x0D; Results: Out of 40 patients, 30 (75%) were males. The average age was 57.78±7.64 years. The p-value of pH and HCO3 was 0.259 and 0.921, respectively. The CO2 in VBGs was 58.10±11.35 mm Hg versus 36.32±6.69 mm Hg in ABG, with a p-value of 0.001, which was statistically significant.&#x0D; Conclusion: In critically ill patients, pH and HCO3 levels of venous blood gases were comparable with ABG. VBG CO2 was significantly higher than ABGs.

Evaluation of a novel heparin-iloprost-based antithrombotic formulation blood collection tube for clinical usage

BackgroundThe objective of our study was to evaluate a single blood collection tube with a novel antithrombotic formulation to measure both hematological, biochemical, and d-dimer analytes. MethodsPaired samples of gold standard blood tubes (EDTA, lithium heparin, sodium citrate) and a new antithrombotic formulation blood tube were collected from 187 patients. The new antithrombotic tube is a lithium heparin tube preloaded with a liquid form of prostacyclin analog. The novel tube was tested on seventeen hematological parameters and smears against EDTA, on fourteen biochemical parameters against lithium heparin and on d-dimer against sodium citrate. ResultsAll correlation coefficients were close to 0.99. The Bland-Altman analyses presented a satisfactory correspondence for all analytes. All the hematological examinations demonstrated comparable results between EDTA and the novel formulation, except for platelet counts analyzed by impedance method, but not by fluorescence. We detected lower mean platelet volume with/without outliers (5.06%)/(5.13%) in the novel formulation and increased mean corpuscular hemoglobin concentration (2.55%). All the biochemistry analytes demonstrated comparable results between lithium heparin and the novel tube. d-dimer showed comparable results between citrated blood and the novel formulation after dilution correction. ConclusionsWe describe a novel antithrombotic formulation tube with the potential to be introduced into clinical laboratories for simultaneous analysis of thirty-two blood analytes.

Prokinetisin 1 preeklampsi belirteci olarak kullanılabilir mi?

Abstract&#x0D; Can prokineticin 1 be used as a biomarker in preeclampsia?&#x0D; Aim:&#x0D; Prokineticins are a group of peptides which play important roles in angiogenesis, hematopoiesis and reproduction. Prokineticin 1 (PROK1) is reported to be the major subgroup affecting the female reproductive development.&#x0D; PROK1 is secreted from the syncytiotrophoblasts and it prevents extravillous trophoblastic cell migration in the first trimester which predispose to the abnormal placentation and preeclampsia (PE). Hypoxia provokes the expression of PROK1 therefore its serum levels increase in pregnants with PE. The aim of the present study was to reveal the value of PROK1 as a marker of PE.&#x0D; Materials and Method:&#x0D; A total of 84 pregnant women were admitted to the study, and 43 of them were diagnosed with preeclampsia. 41 healthy pregnant women were included in the control group. The preeclampsia group was divided into two subgroups as mild PE (n=27) and severe PE (n=16). &#x0D; About 10 ml of venous blood was collected from each participant and dispensed into lithium heparin, and serum was obtained, which were stored at – 80 c until analyzed. Serum PROK1 measurements were performed by using an enzyme-linked immunosorbent assay (ELİSA). All tests were two-tailed, and p

Head-to-Head Comparison of Different Blood Collecting Tubes for Quantification of Alzheimer's Disease Biomarkers in Plasma.

To examine whether the type of blood collection tubes affects the quantification of plasma biomarkers for Alzheimer’s disease analyzed with a single-molecule array (Simoa), we recruited a healthy cohort (n = 34, 11 males, mean age = 28.7 ± 7.55) and collected plasma in the following tubes: dipotassium ethylenediaminetetraacetic acid (K2-EDTA), heparin lithium (Li-Hep), and heparin sodium (Na-Hep). Plasma tau, phosphorylated tau 181 (p-tau181), amyloid β (1–40) (Aβ40), and amyloid β (1–42) (Aβ42) were quantified using Simoa. We compared the value of plasma analytes, as well as the effects of sex on the measurements. We found that plasma collected in Li-Hep and Na-Hep tubes yielded significantly higher tau and p-tau181 levels compared to plasma collected in K2-EDTA tubes from the same person, but there was no difference in the measured values of the Aβ40, Aβ42, and Aβ42/40 ratio. Therefore, the type of blood collecting tubes should be considered when planning studies that measure plasma tau.

Diagnostic potential value of circulating PCA3 mRNA in plasma and urine of prostate cancer patients

Current reports show that most of the routinely available prostate specific antigen (PSA) derived biomarkers of prostate cancer (PCa) have failed to demonstrate sufficient sensitivity and specificity in clinical use for diagnosis and prognosis of PCa. Consequently, a number of molecular markers including the prostate cancer gene 3 (PCA3) are being considered for clinical use. This study was therefore designed to determine the levels of PCA3 mRNA expression in urine and plasma of prostate cancer patients and to assess its diagnostic characteristics in Nigerian population with prostate cancer. A total of 90 participants comprising 30 participants each with PCa, BPH and 30 apparently healthy adults as controls were enrolled into this case-control study. Blood and urine samples were collected from study participants and dispensed into PaxGene RNA protecting fluid, lithium heparin bottles and nuclease free bottle as appropriate. Anthropometric indices were obtained using standard methods, while the plasma levels of PSA, testosterone and estrogen were determined using enzyme linked immunosorbent assay (ELISA). The PCA3 mRNA expression in urine and plasma was determined using Quantitative Polymerase Chain Reaction. Comparison of groups was done using analysis of variance (ANOVA) followed by Tukey Post-Hoc test. The sensitivity and specificity of PCA3 mRNA in the diagnosis of PCa was evaluated through estimation of the area under the receiver operating characteristic (ROC) curve. 'The mean waist circumference (WC) and (HC) was significantly higher in the BPH group (43.94 ± 2.32) compared to both the control (36.04 ± 0.80) and the PCa group (38.50 ± 0.94) (p < 0.05). The prostate cancer group had a significantly higher levels (p < 0.05) of tPSA (47.58 ± 12.23) and an increased expression of PCA3 mRNA in urine (10.46 ± 1.68) and plasma (14.89 ± 1.37) when compared with control values [tPSA (0.67 ± 0.27), urine (2.22 ± 0.22) and plasma (1.45 ± 0.13) expression of PCA3 mRNA]. A negative correlation was found to exist between Plasma PCa3 mRNA and plasma levels of estradiol (r=-0.425, p = 0.024) and testosterone (r=-0.423, p = 0.025) in PCa group. A similar trend was observed between urine PCA3 mRNA, estrogen (r=-0.459, p = 0.014) and testosterone (r=-0.407, p = 0.031). The ROC curve shows the tPSA (p = 0.03) is the most sensitive marker of detection followed by plasma PCA3 mRNA and subsequently the urine PCA3 mRNA. The curve further shows the urine PCA3 mRNA (p = 0.006) is the most specific for detection of prostate cancer and the less and least sensitive are the plasma PCA3 mRNA and tPSA. It could be concluded from this study that there is reduced levels of estradiol and testosterone and elevated levels of tPSA and PCA3 mRNA expression in urine and blood in patients with benign prostatic hyperplasia and prostate cancer. Hence, determination of plasma or urine expression of PCA3 mRNA is valueable in the diagnosis and treatment monitoring of prostate cancer.

Effects of Various Anticoagulants on Biochemistry Analytes From Domestic Pigeons (Columba livia domestica).

The objective of this study was to compare the effects of anticoagulant and no anticoagulant on routine biochemical analytes in domestic pigeons (Columba livia domestica). Blood samples were obtained from 8 clinically healthy pigeons. The sample obtained from each bird was divided into 4 blood collection tubes containing either ethylenediaminetetraacetic acid (EDTA), lithium heparin, sodium citrate, or no anticoagulant. The concentrations of creatinine, uric acid, triglyceride, total cholesterol, glucose, phosphorus, calcium, magnesium, total protein, albumin, and iron, and the activities of alkaline phosphatase (ALP), alanine aminotransferase (ALT), and aspartate aminotransferase (AST), were measured in blood from each of the blood collection tubes. The values of the measured parameters, with the exception of iron, were significantly lower in the citrated plasma samples compared with the serum samples, even after correcting for dilution. In the lithium heparin plasma samples, significant decreases in albumin, triglyceride, calcium, total cholesterol, and ALP, and a significant increase in iron, were observed compared with the values in the serum samples. The concentrations of total protein, creatinine, glucose, total cholesterol, triglyceride, magnesium, calcium, and phosphorus, as well as the activities for AST and ALP, were significantly lower in the EDTA plasma samples compared with the serum samples. In conclusion, the anticoagulants had significant effects on most of the measured parameters compared with serum. The findings of the present study suggest that a lithium heparin sample is the most appropriate plasma sample for the measurement of blood biochemical parameters in the domestic pigeon.

Estimation of Homocysteine Level in Sudanese Obese, Gezira State, Sudan

Science-based evidence on the association of total homocysteine (Hcy) with body mass index (BMI), as well as obesity and elevated Hcy levels, in the general population has been conflicted .The primary goals of this case control study is to evaluate the relationship between Hcy concentration and obesity in Gezira state in Sudan. A total of 221 participants including 140 obese&nbsp;,&nbsp;their average age was 49.49±12.2 years,&nbsp;divided into three groups on the basis of BMI according to the WHO classification and the groups constituted 44 obese type I&nbsp;,&nbsp;51 obese type II and 45 obese type III&nbsp;, the control group was normal weight (BMI &lt; 25.9)&nbsp;&nbsp; include 81 participants ,their age medium were 45.78±22.38 years .Three mL of venous blood from each participant was collected in the morning after a twelve hours overnight fasting in lithium heparin container, then the separated plasma was stored at −80 ◦C for further analysis. Serum Hcy was measured using the enzymatic method by Cobas C 411analyzer. The cut-off value for normal levels of Hcy is 15 μmol/L , out of 140 obese, there were 43(30.7%) having hyperhomocysteinemia (HHcy).The Hcy average increased in obese (12.65±4.9) in comparing with control(10.45±3.8) giving significant difference (P value 0.002).&nbsp; The mean values of Hcy were also compared among the various obesity types. It was found that there was no statistically significant (P value0.628). Pearson correlation analyses showed that BMI was negatively associated with Hcy (r-0.027, P value 0.754). The mean of Hcy was decreased in the lower&nbsp; age group (20 – 40 years), and increased in the higher age group (61 – 80years) showing no significant difference but age is positively correlated with high level of Hcy (r 0.094, P value: 0.268). The study finding showed significant relationship between Hcy and obesity. However, the Hcy concentrations were not positively correlated with the degree of obesity, further studies are needed to ascertain the causal role of Hcy in the development of obesity and to explore how Hcy is involved in the pathogenesis of obesity.

Resolving icteric interference by a novel photoisomerization device: Bilibox

Icterus, a phenomenon caused by bilirubin elevation in the blood, is a common endogenous interference in chemistry testing, occurring either spectrally or through chemical reactivity with assay reagents. Often, laboratories have few options other than to dilute or reject samples exceeding icteric thresholds. However, recent studies have optimized in vitro photoisomerization of bilirubin to a 17-minute bilirubin half-life using 500 nm light at 37 °C. Using an enzymatic creatinine assay as a model, due to its prevalence in routine laboratory testing and susceptibility to icteric interference, our study explores the usage of in vitro photoisomerization by replicating these conditions in a device, the Bilibox, as means of resolving icterus in laboratory testing.Left-over icteric and non-icteric clinical samples, collected by lithium heparin vacutainer (n = 10), were analyzed for baseline creatinine, diluted creatinine (1:4 0.9 % NaCl), total bilirubin, direct bilirubin, and hemolysis, icterus and lipemia (HIL) indices. Samples were then placed in the Bilibox in two intervals of 30 min with repeat measurements of the aforementioned analytes.On average, icteric-index, total bilirubin (TBIL), and direct bilirubin (DBIL) decreased by 33.5, 39.1 and 39.9 % respectively following 30 min of Bilibox treatment; and by 47.6 %, 63.7 % and 59.8 % following 60 min. The average percent difference between the pre-exposure diluted and undiluted creatinine was 5.8 %, demonstrating the icterus interference. Following Bilibox treatment, the difference between undiluted (post-exposure) and diluted (pre-exposure) creatinine decreased to 0.02 % (p = 0.0232) and 2.2 % (p = 0.0021) at 30 and 60 min of treatment respectively, demonstrating resolution of interference. Consequently, photoisomerization can be utilized as an additional and reasonably quick method for resolving icterus when dilutional methods cannot be applied.

Simple, high-throughput measurement of gut-derived short-chain fatty acids in clinically relevant biofluids using gas chromatography-mass spectrometry

IntroductionThe quantitative measurement of circulating gut bacteria-derived metabolites has increased in recent years due to their associations with health and disease. While much of the previous attention has been placed on metabolites considered as deleterious to health, a shift to the investigation of short-chain fatty acids (SCFAs) as potential health promotors has been observed. ObjectivesTo develop a simple, high-throughput and quantitative assay to measure gut-derived SCFAs in clinically relevant biofluids using gas chromatography-mass spectrometry (GC–MS). MethodsA short (7.5 min) GC–MS assay was optimized for measurement of seven straight- and branched-chain SCFAs and their deuterated isotopes using a wax-based column for analysis without prior derivatization. The assay was validated using routine criteria to assess precision, accuracy, matrix effects, recovery, and extraction reproducibility. Assay applicability was tested in cohorts of healthy individuals and kidney disease patients. ResultsThe assay was demonstrated to be precise, accurate and reproducible with acceptable levels of matrix effect and analyte recovery. Lower limits of detection and quantitation were in the low ng/mL range. An investigation into different blood collection tube chemistries demonstrated that lithium heparin plasma and serum clotting activator tubes are recommended for use in future cross-study comparisons. Kidney disease patient analyses demonstrated variable differences across SCFAs when comparing hemodialysis to earlier stages of chronic kidney disease, demonstrating the suitability of the assay for translation to clinical analyses. ConclusionThe assay has been validated and identified as reliable for use in larger-scale studies for the analysis of SCFAs in human plasma and serum.

Adult and neonate TSH blood spot reference intervals.

Background: Neonatal congenital hypothyroidism screening is performed by measuring thyroid-stimulating hormone (TSH) in a dried blood spot (DBS) sample, whereas acquired hypothyroidism uses serum TSH. There is no established DBS TSH reference interval, but knowing this is useful, as some patients cannot tolerate venepuncture, so DBS collection is seen as an acceptable alternative. The aim of this study was to establish DBS TSH reference intervals in adults and neonates (day 5-8), and determine the relationship between serum and DBS TSH.Methods: Euthyroid adults, not on thyroid medication and with a normal haematocrit, were selected. If they had a paired lithium heparin sample, DBS were prepared by spotting 50µL of whole blood onto filter paper. Dried blood spot TSH was measured using the PerkinElmer Neonatal hTSH kit on the GSP instrument and serum using the Abbott Architect assay. The relationship between DBS and serum TSH was analysed using Passing-Bablok regression and the adult DBS TSH reference interval calculated using transformed data. The neonatal reference interval was calculated from screening results using the non-parametric method.Results: Overall, 109 adult samples were included in the study (61 female). The Passing-Bablok regression was: DBS TSH = 0.68 × serum TSH + 0.07, and reference interval was 0.49-3.07 mU/L. The neonatal DBS reference interval was 0.40-4.10 mU/L from 8351 results.Conclusion: This study derived adult and neonate TSH DBS reference intervals using the GSP analyser and established the relationship between serum and DBS TSH. Knowing this information will allow for improved interpretation of DBS TSH results.

Association of Biochemical Parameters in Chronic Drug Addict Patients; A Case-Control Study

Objective: To determine the effect of a biochemical parameter in chronic drug addict patients in Peshawar, Khyber Pukhtunkhwa. Materials and Methods: Blood samples of 90 drug addicts were randomly collected, 45 were hashish and 45 were heroin addicts. Drug addicts having hepatitis B and C infections were excluded from the study. Three milliliters of blood samples were collected in the lithium heparin tube using an aseptic technique for the determination of Alanine transaminase, Aspartate transaminase, Triglycerides, and Cholesterol. Results: Cholesterol was significantly decreased (P=0.00) in the age group between 21 and 40 years followed by the age group less than 20 years (P=0.02). Liver enzymes were significantly elevated but, were more significant up to 40 years (P=0.00). Conclusion: Heroin and hashish affect liver function enzymes Alanine transaminase and aspartate transaminase and cause a reduction in cholesterol levels in individuals using drugs for a long period. Keywords: Alanine transaminase (ALT), Aspartate transaminase (AST), Triglycerides, and Cholesterol.

SHORT-TERM BIOLOGICAL VARIABILITY OF HEMATOLOGY PARAMETERS IN THE BEARDED DRAGON (POGONA VITTICEPS)

Determination of the biological variation of laboratory tests is essential for accurate interpretation during health assessments. Few studies investigate the biological variation of CBC variables in reptiles, and none of these involve squamates. Thus, we investigated the biological variation of hematology parameters in bearded dragons (Pogona vitticeps) to determine if conventional population-based reference intervals are appropriate in evaluating single laboratory samples. Blood was collected from 10 bearded dragons five separate times over 10 wk and placed in lithium heparin (LH) microtainers, and CBC were performed. WBC were evaluated via both a fresh, non-anticoagulated blood smear and a LH anticoagulated blood smear as well as two manual hemocytometer counts with blood stained with either Natt and Herrick's (NH) solution or Leukopet™ (LO) solution. The between-animal coefficient of variation (CVG), within-animal coefficient of variation (CVI), reference change value (RCV), and index of individuality were determined for PCV, total solids (TS), and WBC using all leukocyte quantification methods. The RVC for PCV, TS, and WBC in bearded dragons were 37%, 84%, and >100%, respectively. The calculated index of individuality values all fell between 0.6 and 1.4, suggesting that the use of judiciously applied population-based reference intervals in interpreting the WBC, PCV, and TS in bearded dragons is appropriate.

Specific features of blood parameters in volleyball players and wrestlers in preparatory period of training cycle

Annotation. Study of modern laboratory markers of structural and functional disturbances of muscle tissue in athletes, reflecting energy metabolism, paravertebral muscle damage, and being an indicator of body performance and muscle activity, is of undeniable practical importance for modern sports medicine. The aim of the study was to determine clinical and biochemical parameters of blood in volleyball players and wrestlers in assessment of structural and functional changes in skeletal muscles. Blood examination was carried out in 26 volleyball players and 25 middleweight Greco-Roman wrestlers as part of repeated comprehensive medical examination being conducted at the Department of Physical Education of Vinnytsia National Pirogov Memorial Medical University. Eligible subjects included athletes 17 to 21years of age having first adult category to master of sports and being in preparatory period of annual training macrocycle. The athletes were examined in the morning, on empty stomach, not less than 12 hours after training. The control group consisted of 25 practically healthy students having moderate physical activity. Clinical blood indices were determined by conductometric method on an automatic hematology analyzer ABX HORIBA PENTRA 60 C + (France). Hormonal studies were carried out by immunochemiluminescence method on automatic analyzer “ACCESS-2”, Bekchman Coulter (USA). Biochemical studies were performed using an automatic analyzer AU-480, Bekchman Coulter (USA). Electrolyte content was determined by ion-selective electrode technology on Medica electrolyte analyzer in EasyElectrolytes™, using lithium heparin vacuum system. Glucose levels were determined on automatic analyzer Biosen (Germany). Statistical processing was done using the program “Statistica 5.5”. Significance of differences between the variables was determined by Mann-Whitney U-test. The following serum humoral factors were found to be of great significance in assessment of structural and functional changes in skeletal muscles in volleyball players and wrestlers: the number of large immature cells of monocytes and platelets, electrolyte content, concentration of creatine phosphokinase and lactate dehydrogenase, creatinine level, as well as triglycerides and lactate levels. Establishing blood biomarkers should be an integral part of scientific and practical monitoring of health status in team athletes and wrestlers.