Laboratory medicine and clinical medicine are co-dependent components of medicine. Laboratory medicine functions most effectively when focused through a clinical lens. Me dical practice as a whole undergoes change. New drugs, treatments and changes in management strategies are introduced. New techniques, new technologies and new tests are developed. These changes may be either clinically or laboratory initiated, and so their introduction requires dialogue and interaction between clinical and laboratory medicine specialists. Treatment monitoring is integral to laboratory medicine, varying from direct drug measurement to monitoring cholesterol levels in response to treatment. The current trend to »personalised medicine« is an extension of this process with the development of companion diagnostics. Technological innovation forms part of modern laboratory practice. Introduction of new technology both facilitates standard laboratory approaches and permits introduction of new tests and testing strategies previously confined to the research laboratory only. The revolution in cardiac biomarker testing has been largely a laboratory led change. Flexibility in service provision in response to changing clinical practice or evolving technology provides a significant laboratory management challenge in the light of increasing expectations, shifts in population demographics and constraint in resource availability. Laboratory medicine practitioners are adept at meeting these challenges. One thing remains constant, that there will be a constant need laboratory medicine to meet the challenges of novel clinical challenges from infectious diseases to medical conditions developing from lifestyle and longevity.
Diagnostic blood samples collected by phlebotomy are the most common type of biological specimens drawn and sent to laboratory medicine facilities for being analyzed, thus supporting caring physicians in patient diagnosis, follow-up and/or therapeutic monitoring. Phlebotomy, a relatively invasive medical procedure, is indeed critical for the downstream procedures accomplished either in the analytical phase made in the laboratory or in the interpretive process done by the physicians. Diagnosis, man agement, treatment of patients and ultimately patient safety itself can be compromised by poor phlebotomy quality. We have read with interest a recent article where the authors addressed important aspects of venous blood collection for laboratory medicine analysis. The authors conducted a phlebotomy survey based on the Clinical and Laboratory Standard Institute (CLSI) H03-A6 document (presently replaced by the GP41-A6 document) in three government hospitals in Ethiopia to evaluate 120 professionals (101 non-laboratory professionals vs. 19 laboratory professionals) as regards the venous blood collection practice. The aim of this mini (non-systematic) review is to both take a cue from the above article and from current practices we had already observed in other laboratory settings, and discuss four questionable activities performed by health care professionals during venous blood collection. We refer to: i) diet restriction assessment; ii) puncture site cleansing; iii) timing of tourniquet removal and; iv) mixing specimen with additives.
Working in laboratories of clinical chemistry, we risk feeling that our personal contribution to quality is small and that statistical models and manufacturers play the major roles. It is seldom sufficiently acknowledged that personal knowledge, skills and common sense are crucial for quality assurance in the interest of patients. The employees, environment and procedures inherent to the laboratory including its interactions with the clients are crucial for the overall result of the total testing chain. As the measurement systems, reagents and procedures are gradually improved, work on the preanalytical, postanalytical and clinical phases is likely to pay the most substantial dividends in accomplishing further quality improvements. This means changing attitudes and behaviour, especially of the users of the laboratory. It requires understanding people and how to engage them in joint improvement processes. We need to use our knowledge and common sense expanded with new skills e.g. from the humanities, management, business and change sciences in order to bring this about together with the users of the laboratory.
Hemoglobin A1c, (HbA1c) which is the major constituent of glycated hemoglobin, has been used in the follow-up of retrospective glycemia for years and in the diagnosis of diabetes mellitus nowadays. Since the analytical performance of HbA1c should be high likewise all laboratory tests, various quality control measures are used. Sigma metrics is one of these measures and it is the combination of bias, precision and total allowable error that ensures a general evaluation of analytical quality. The aim of our study was to evaluate the analytical performance of Bio-Rad’s Variant Turbo II HbA1c analyzer according to sigma metrics. Sigma levels were calculated using the data obtained from two levels of internal and 12 external quality control materials (Bio-Rad) of Variant II Turbo HbA1c analyzer according to s= (TEa% - Bias%) / CV% formula. The mean sigma levels for low and high quality control materials were found to be 3.0 and 4.1, respectively. The annual mean analytical performance of Variant II Turbo HbA1c analyzer was found to be acceptable according to sigma metrics. In order to be sure of the difference in HbA1c results indicating the success or failure in treatment but not arise from analytical variation, it is thought that more stringent quality control measures should be applied to reach higher sigma levels.
Background: Vascular calcification (VC) is highly prevalent in dialysis (HD) patients, and its mechanism is multi factorial. Most likely that systemic or local inhibitory factor is overwhelmed by promoters of VC in these patients. VC increased arterial stiffness, and left ventricular hypertrophy. Thus, the present study aimed to investigate the association of VC and myocardial remodeling and to analyze their relationship with VC promoters (fibroblast growth factor 23-FGF23, Klotho, intact parathormon-iPTH, vitamin D) in 56 prevalent HD patients (median values: age 54 yrs, HD vintage 82 months). Methods: Besides routine laboratory analyzes, serum levels of FGF 23, soluble Klotho, iPTH, 1,25-dihydroxyvitamin D3; pulse wave velocity (PWV); left ventricular (LV) mass by ultrasound; and VCs score by Adragao method were measured. Results: VC was found in 60% and LV concentric or eccentric hypertrophy in 50% patients. Dialysis vintage (OR 1.025, 95%CI 1.007-1.044, p=0.006) FGF23 (OR 1.006, 95% CI 0.992-1.012, p=0.029) and serum magnesium (OR 0.000, 95%CI 0.000-0.214, p=0.04) were associated with VC. Changes in myocardial geometry was associated with male sex (beta=-0.273, 95% CI -23.967 1.513, p=0.027), iPTH (beta 0.029, 95% CI -0.059-0.001, p=0.027) and vitamin D treatment (beta 25.49, 95% CI 11.325-39.667, p=0.001). Also, patients with the more widespread VC had the highest LV remodeling categories. PWV was associated patient's age, cholesterol, diastolic blood pressure, LV mass (positively) and serum calcium (negatively), indicating potential link with atherosclerotic risk. Conclusions: Despite to different risk factors for VC and myocardial remodeling, obtained results could indicate that risk factors intertwine in long-term treatment of HD patients and therefore careful and continuous correction of mineral metabolism disorders is undoubtedly of the utmost importance.
Background: Diabetes mellitus (DM) with its micro- and macrocomplications is the leading global epidemic of the 21st century. The aim of the research is to determine possible changes in the complete blood count (CBC) parameters depending on glycemic controlin patients with Type 2 diabetes mellitus (T2DM). Methods: The study included a total of 178 patients with T2DM, both gender over the age of 40 years, from the Health Care Center "Dr Milorad Mika Pavlovic" Indjija, Serbia. To notice the possible correlation between the CBC parameters and glucose control in T2DM, the subjects were divided in two groups with HbA1c 7%. We analysed CBC parameters, parameters of glycoregulation, lipid status using standard biochemical methods, performed anthropometric measurements and collected patients data by questionnaire and electronic patient card. Results: There was statistical difference between HbA1c groups for PMDW (p=0.045), HDL (p=0.0067). Using univariate linear regression it is shown that PCT was correlated with WBC (p=0.0005), neutrophils (p=0.046), monocytes (p=0.003); MPM was associated with MPV (p=0.0005); MPC (p=0.0005), PDW (P=0.0005), GLU0 (p=0.034), HDL-C (p=0.005); PMDW was correlated with HbA1c% (p=0.049), GLU0 (p=0.013), HDL-C (p=0.001), BW (p=0.043) in all patients. Conclusions: Based on our study results it may be concluded that some of the parameters of CBC could be useful tool in following glycemic control of diabetics.
Background: FTO, a gene recently discovered in genome-wide associated studies for type 2 diabetes mellitus (T2D), play an important role in the management of energy homeostasis, nucleic acid demethylation and regulation of body fat mass by lipolysis. The aim of this study was to analyze the association of FTO rs8050136 A>C genetic variant with clinical and biochemical parameters of T2D in the population of West Balkan region (Bosnians and Herzegovinians and Kosovars). Methods: The study included 638 patients with T2D and prediabetes and 360 healthy controls of both genders, aged from 40 to 65 years. Patients were recruited at the Clinical Centre University of Sarajevo, University Hospital of Clinical Centre in Banja Luka, General Hospital in Tesanj and Health Centre in Prizren. Genotyping of analyzed FTO polymorphism rs8050136 A>C was performed by qPCR allelic discrimination. Results: Genotype frequencies of the analyzed polymorphism were comparable between patients with T2D, prediabetic patients, and healthy population. Logistic regression analyses didn't show significant association of FTO rs8050136 A allele with increased risk of T2D. However, risk A allele was significantly associated with higher levels of HbA1c, insulin, HOMA-IR index, diastolic blood pressure, and inflammatory markers (fibrinogen and leukocytes) as well as showed tendency of association with increased values of obesity markers (BMI, waist and hip circumference). Conclusions: Results of our study showed a significant association of FTO genetic variant rs8050136 A>C with the major markers of insulin resistance, obesity and inflammation, opening new avenues for solving many unclear questions in the pathogenesis of T2D.
Background: C-Myc is one of the major cellular oncogenes overexpressed in non-small cell lung carcinoma (NSCLC). Its deregulated expression is necessary but not sufficient for malignant transformation. We evaluated expression of MYC gene in NSCLC patients and its association with alterations in the genes previously identified to be related to NSCLC pathogenesis, PHACTR3 and E2F4. Methods: We analyzed MYC gene expression by qRT-PCR in 30 NSCLC patients' samples and paired normal lung tissue. MYC expression was further statistically evaluated in relation to histopathological parameters, PHACTR3 and E2F4 gene alterations and survival. Alterations in aforementioned genes were previously detected and identified based on AP-PCR profiles of paired normal and tumor DNA samples, selection of DNA bands with altered mobility in tumor samples and their characterization by the reamplification, cloning and sequencing. Results: MYC expression was significantly increased in NSCLC samples and its overexpression significantly associated with squamous cell carcinoma subtype. Most importantly, MYC overexpression significantly coincided with mutations in PHACTR3 and E2F4 genes, in group of all patients and in squamous cell carcinoma subtype. More over, patients with jointly overexpressed MYC and altered PHACTR3 or E2F4 showed trend of shorter survival. Conclusions: Overall, MYC is frequently overexpressed in NSCLC and it is associated with mutated PHACTR3 gene, as well as mutated E2F4 gene. These joint gene alterations could be considered as potential molecular markers of NSCLC and its specific subtypes.
Background: Cardiac surgery-associated acute kidney injury (CSA-AKI) frequently occurs in patients assessed as low-risk for developing CSA-AKI. Neutrophil Gelatinase-Associated Lipocalin (NGAL), Kidney Injury Molecule-1 (KIM-1) and lactate are promising biomarkers of CSA-AKI but have not yet been explored in low-risk patients. Aim: To evaluate urinary NGAL (uNGAL), KIM-1 and lactate as biomarkers of CSA-AKI in patients with low-risk for developing CSA-AKI. Methods: This prospective, observational study included 100 adult elective cardiac surgery patients assessed as low-risk for developing CSA-AKI. UNGAL, KIM-1 and lactate were measured preoperatively, at the end of cardiopulmonary bypass (CPB) and 3, 12, 24 and 48 h later. Results: Fifteen patients developed CSA-AKI. Patients with CSA-AKI had significantly higher lactate but similar uNGAL and KIM-1 levels compared to patients without CSA-AKI. Unlike uNGAL and KIM-1, postoperative lactate was good biomarker of CSA-AKI with the highest odds ratio (OR) 2.7 [1.4-4.9] 24 h after CPB. Peak lactate concentration >= 4 mmol/L carried dramatically higher risk for developing CSA-AKI (OR 6.3 [1.9-20.5]). Conclusions: Unlike uNGAL and KIM-1, postoperative lactate was significant independent predictor of CSA-AKI with the highest odds ratio 24 h after CPB.
During (April 11 and 12, 2019, Belgrade, ) Society of Medical Biochemists of Serbia organized scientific and professional program with aim to discuss laboratory medicine topics of mutual interest for all the countries of the Region, such as: Together with the countries from Balkan Region the countries from our neighborhood as Italy, Austria, Hungary, Cyprus and Israel have been invited to discuss this important topics and exchange the mutual experiances with aim to improve the laboratory medicine in our countries and to help our colleagues to improve daily laboratory practice in our countries. Also participation in the Symposium took colleagues from France and Belgium.
Background: We analyzed cardiovascular inflammatory (C-reactive protein (CRP), interleukin 6 (IL-6)), haemostatic (homocysteine) risk markers in lean and obese patients at admission and acute hyperglicemic crisis (AHC) resolving, involving diabetic ketoacidosis (DKA) and hyperosmolar hyperglycemic state (HHS). Methods: In that context, we included group A: N=20 obese, B: N=20 lean patients with DKA; C: N=10 obese, D: N=10 lean patients with HHS; E: N=15 obese, F: N=15 lean controls. CRP, IL-6, homocysteine were determined by ELISA. Results: Our results showed that CRP, IL-6, and homo cysteine levels decreased in all groups: (A: p<0.001; B: p<0.001, C: p<0.05; D: p<0.001 mg/L), (A: p<0.001 B: p<0.001, C: p<0.001, D: p<0.01 pg/mL), (A: p<0.001, B: p<0.001; C: p<0.05, D: p=0.001 mmol/L), respectively, at resolving AHC. However, CRP persisted higher (p<0.001, p<0.01), IL-6 lower (p<0.05, p<0.001), while homo cysteine levels turned out to be similar to controls. Conclusions: AHC is associated with increased inflammatory and hemostatic cardiovascular risk markers. Also, insulin therapy in AHC has had more pronounced favorable effect on IL-6 and homocystein than on CRP.
Background: Vitamin D regulates calcium and phosphorus metabolism, and it is essential for bone formation. Several factors can affect vitamin D levels in plasma. In present study we compare vitamin D levels of outpatients, who admit to Maltepe University Hospital between 2011 and 2013 and had vitamin D measurements regarding gender, age, and season. Methods: Hospital records were evaluated to identify the outpatients with vitamin D levels and their gender, age, and vitamin D levels and the seasons of measurements were recorded. Results: Data of 4860 subjects (74% female) were analyzed and 69.2% were between 18-64 years old. Vitamin D levels were as follows: 43.1% = 30 ng/mL. The number of females with vitamin D levels 30 ng/mL. Conclusions: The prevalence of vitamin D deficiency in outpatients of Maltepe University Hospital in Marmara region was 75% (< 20 ng/mL).
Background: To date few reports have pointed out the role of circulating miRNAs in discriminating metastatic liver tumors from primary hepatocellular (HCC) tumors. Such discrimination will have significant therapeutic and prognostic implications. The purpose of this study was to evaluate the potential value of a panel of HCC-related circulating miRNAs (miR-142, miR-182, miR-200a, mir-210, miR-211, miR-302b, miR-324, miR-338, miR-340 and miR-1246) as noninvasive biomarkers for discriminating primary HCC from metastatic tumors in the liver. Methods: The expression level of the selected miRNAs was quantified by quantitative real time PCR in 33 patients with HCC, 22 patients with metastatic tumors in the liver, and 30 healthy volunteers as control. Mann-Whitney U test was used to evaluate the difference in miRNAs expression between primary and metastatic liver tumors and to study the associations between their relative expression levels and the clinicopathological factors. Receiver operating characteristic curve was used to evaluate the diagnostic value of the individual miRNAs. Results: Statistical analyses revealed a differential expression in the level of serum miR-210 and miR-1246 between the two groups of patients. The sensitivity and specificity of miR-210, for differentiating HCC from metastatic malignancies in the liver were found to be 73.7% and 64.28%, respectively. Whilst, of miR-1246 were 72.2% and 67.8%, respectively. In addition, the differential expression of the two miRNAs was also found to be associated with clinicopathological parameters in the two studied groups. Conclusions: Serum miR-210 and miR-1246 have some diagnostic value for discriminating patients with metastatic tumors to patients with primary HCC
Background: Although vitamin D in not a traditional marker for cardiovascular and renal diseases, several studies have proposed a correlation between vitamin D deficiency and these diseases due to the effect of vitamin D on endothelial function. Asymmetric and symmetric dimethyl arginine (ADMA and SDMA, respectively) are endogenous markers of endothelial dysfunction, and are considered as future markers for the assessment of cardiovascular and renal diseases. The present study investigated the association of kidney function tests (urea and creatinine) and dimethylarginine toxins (ADMA and SDMA) in women with vitamin D insufficiency or deficiency. Indeed, sex hormones (estrogen and testosterone) were analyzed in the participants. Methods: Women were divided into two groups: premenopausal women (younger than 50 years) and postmenopausal women (older than 50 years). Urea, creatinine, estrogen, testosterone, ADMA, and SDMA levels were analyzed when vitamin D level was deficient or insufficient in the participants. Results: The premenopausal women group showed no significant correlations between dimethylarginine toxins and renal failure tests or sex hormones. In the elderly (postmenstrual) women group, only SDMA was significantly correlated with urea and creatinine, while both ADMA and SDMA were not correlated with sex hormones. Conclusions: Although ADMA and SDMA are promising candidates of endothelial dysfunction and are increased in menopause and aging, no direct link between ADMA and further progression of renal failure was observed in women with low vitamin D levels. In contrast, a possible direct correlation between SDMA and renal dysfunction was noticed, but only in an age-dependent manner.
Background: To establish a fast and simple quantitative method for detection of heart-type fatty acid-binding protein (H-FABP) in serum based on a background fluorescence quenching immunochromatographic assay. Methods: A detection card based on the double-antibody sandwich double-antibody method with background fluorescence quenching was developed for quantitative measurement of H-FABP in serum. The optimal concentrations of control for coating the test and control lines were determined as well as the concentrations of gold-labeled antibodies used in preparing the detection system. The detection method for H-FABP in serum was established and validated using real-world clinical samples. Results: The optimal concentrations of labeling antibody and coating antibody were 5.0 mu g/mL and 1.0 mg/mL, respectively. The test card had a sensitivity of 1.15 ng/mL over a linear concentration range of 0-100 ng/mL. Based on three batches prepared for testing the card, the relative standard deviation (RSD) within batches was less than 15% without a significant difference (P=0.942). The detection method was tested against common interfering substances in serum, such as bilirubin, triglyceride and serum anticoagulants ethylenediamine tetraacetic acid (EDTA), heparin, and sodium citrate, and no significant cross-reaction was detected. The test method was further validated with 50 clinical serum samples, and the test results were comparable with standard reference detection methods with good correlation (R=0.95). Conclusion: Our study presents a new method with strong specificity and sensitivity for the detection of H-FABP in serum, which could promote H-FABP detection in a broad range of applications.
Mutations in BRCA1 gene have been implicated in ovarian cancers, and BRCA testing may be conducted in high-risk women. This study was designed to determine the frequency of three single nucleotide polymorphisms (SNPs) variants in BRCA1 gene and BRCA1 expression in Saudi females with ovarian cancer. Expression levels of mRNA of BRCA1 gene were studied in 10 ovarian cancer and 10 normal ovarian tissues, by quantitative real time polymerase chain reaction (qPCR). The study also included 28 females who had suffered from ovarian cancer and had been successfully operated upon and 90 healthy females with no history of cancer. Blood was drawn in EDTA tubes and used for extraction of DNA. The genotyping was carried out using Taqman® SNP Genotyping kit by RT-PCR. The variants investigated included c.871 T>C (rs799917), c.1040 G>A (rs4986852), c.181 T>G (rs28897672) in BRCA1 gene. The c.181 T>G (rs28897672) showed significantly different genotype and allele frequencies between the patients and the control subjects (p value = 0.002 and 0.02, respectively). The genotype TG was significantly protective (OR = 0.36, p value = 0.024). The mRNA expression of BRCA1 gene was found to be low in the ovarian cancer tissues. This study showed that c.181 T>G in BRCA1 genes is associated with the development of ovarian cancer in Saudis. More studies are needed to unveil other SNPs that may be associated with ovarian cancer and to understand the mechanism(s) involved in reducing the expression of BRCA1 gene in ovarian cancer tissues.
The aim of the study was to investigate the association of Cystatin C (CysC) to biochemical markers of bone turnover and bone mass, and to evaluate its prognostic significance in elderly males with chronic heart failure (CHF). A prospective cohort study was executed on sixtyeight males (mean age 68±7 years) with mild to moderate CHF, together with 19 of corresponding age- and body mass index-matched healthy individuals who underwent cardio vascular, bone mineral density (BMD), and body com position assessment. Biochemical assessment of all subjects included NT-pro-BNP, parathyroid hormone (PTH), 25-hydroxy vitamin D (25(OH)D), CysC, and biochemical markers of bone turnover including osteocalcin (OC), alkaline phosphatase (ALP), β-CrossLaps (β-CTx), osteoprotegerin (OPG), and receptor activator of nuclear factor κB ligand (RANKL). Serum CysC was significantly increased in males with CHF in comparison to healthy control ones. A significant positive association was found between CysC levels and OC in males with CHF, while OC and β-CTx increased in increasing CysC tertiles. In multivariate regression analysis, OC and smoking were a significant determinant of CysC in males with CHF. Level of CysC was found to be positively associated with an increased fatal risk in males with CHF. Serum osteocalcin is an independent predictor of CysC level in elderly males with CHF. Higher CysC level showed a negative relation to survival and bone loss in males with CHF. Further research is needed to confirm the potential role of CysC in the crosstalk between heart, kidney, bone, and energy metabolism in CHF.
Endocrine system plays a major role in both permissive and regulatory activities in order to adequately respond to physical stress of exercise. But level and direction of activation depend on many factors and are not easily interpreted. We tested a group of male professional athletes (21 water polo players and 15 wrestlers), together with 20 sedentary controls matched by age. All participants took a continuous progressive exercise stress test on a treadmill until exhaustion and plateau of oxygen consumption (VO ). Blood samples for cortisol, sex hormone binding globulin (SHBG) and testosterone were drawn in four time points: baseline (B), start of the test (S), point of maximal strain (MAX) and in the 3 minute of recovery period (R). Cortisol levels significantly increased in both groups, but the response between S and MAX was more pronounced in controls (p=0.036). The athletes had significantly higher levels of cortisol in all points in test, except during R (p=0.118), when their cortisol levels gradually started to decline. Significant increase in total testosterone was in great deal a consequence of increase in SHBG level (p<0.01 for both). Consequently, calculated free testosterone significantly decreased during test (p=0.008), and the drop was more pronounced in athletes. This was in concordance with significant correlation between SHBG and cortisol level demonstrated in athletes, but not in controls. It seems that high intensity endurance exercise favors catabolic response, but the level of response highly depends on a previous level of training.
Triple negative breast cancer (TNBC) is characterized by aggressive clinical course and is unresponsive to anti-HER2 and endocrine therapy. TNBC is difficult to treat and is often lethal. Given the need to find new targets for therapy we explored clinicopathological significance of copy number gain of and . Our aim was to determine the impact of and copy number gain on clinical course and outcome of TNBC. and gene copy number alterations were evaluated in 78 archive TNBC samples using TaqMan based quantitative real time PCR assays. 50% of samples had increased copy number. copy number gain was associated with TNBC in contrast to ER positive cancers. Our results showed significant correlation between copy number gain and high grade of TNBCs. This suggests that copy number could be an useful prognostic marker for TNBC patients. copy number gain was associated with high pTNM stage as well as lobular and medullary tumor subtypes. 43% of samples had increased copy number. No correlations between copy number gain and clinicopathological variables were observed. We identified copy number gain as a prognostic marker for TNBC. Our results indicate that may contribute to TNBC progression. We observed no significant association between and/or copy number status and patient survival.
Neopterin is a pyrazino-pyrimidine compound which is used as a marker of cell-mediated immunity in a variety of diseases. It is known that neopterin levels increase in diseases where interferon-gamma (IFN-g) stimulation is present, and also as a result of deficiencies in renal function, given that the primary means of elimination of neopterin is through the kidneys. In this study, we aimed to investigate the role of increased neopterin levels as a prognostic biomarker in patients with impaired renal function. A total of 90 individuals including 63 patients with chronic kidney failure (CKF) and 27 healthy volunteers were included in the study. Serum neopterin concentrations were measured using the enzyme-linked immunosorbent assay. A Mann-Whitney U test and a Pearson Correlation Test were used in the statistical analysis, with a value of 0.05). A significant increase was found in the serum neopterin levels in the CKF patients, due to both the triggering of the disease and the reduction of neopterin elimination.