Assessing post-analytical phase harmonization in European laboratories: a survey promoted by the EFLM Working Group on Harmonization.

OBJECTIVES: Harmonization of the laboratory total testing process (TTP) is critical to improving patient outcome. In 2016, an EFLM survey on the harmonization of TTP underlined the serious shortcomings pertaining to the post-analytical phase. In 2023, the WG-H conducted a new survey aiming to update information in the 2016 harmonization report in order to ascertain whether countries that had declared they were keen to adopt SI units had continued with this program, the aim being to verify the state-of art in harmonization units in areas of laboratory medicine not included in the previous survey. METHODS: Questionnaires were distributed to the Presidents and National Representatives of EFLM Full Member Societies and EFLM affiliate Members. The survey questions were grouped into three categories: measurement units, reference intervals, and nomenclature/terminology, and results were evaluated using Survey Monkey software and Excel. RESULTS: A total of 123 questionnaires from 31 countries were analyzed. A trend (+19.3 %) was observed toward a wider use of SI units for general clinical biochemistry parameters. The results for tests not included in the 2016 survey (i.e., endocrinology diagnostics and coagulation panels), demonstrated that for reports on hormones, responses were satisfactory, 70-90 % of the responders adopting the recommended units, whereas for coagulation test panels, a serious lack of harmonization was found, "seconds", which are inaccurate and not recommended, being widely used units (91 %). CONCLUSIONS: The findings made in the 2023 survey demonstrated a progressive, albeit slow, improvement in harmonization reports. However, further efforts at improvement are mandatory.

Verification of bile acid determination method and establishing reference intervals for biochemical and haematological parameters in third-trimester pregnant women.

OBJECTIVES: The aims of this study were to verify the bile acids (BA) method and to establish reference intervals (RIs) for bile acids (BA) and biochemical and haematological parameters in Croatian pregnant women. METHODS: BA spectrophotometric method verification was performed on Siemens Atellica Solution CH 930 automated analyser using Sentinel reagent. Stability, precision, trueness, linearity, and RIs, as well as lipemia interference were tested according to CLSI guidelines. BA, biochemical, and haematological parameters were measured in serum (BA, biochemical) and whole blood (haematological) samples of fasting healthy third-trimester pregnant women from Croatia (n=121). The establishment of the RIs was done a priori according to the CLSI EP28-A3C:2010 guideline. Selected reference individuals' data were analysed using parametric, non-parametric, and robust methods. RESULTS: Stability study showed that BA are stable in serum samples for 2 days at 20 °C, 14 days at 4-8 °C, and 22 days at -20 °C. The precision study and adult RIs verification met the criteria. Linearity was verified for the concentration range of 3.5-172.1 µmol/L whereas the lipemia interference test showed a positive bias (%) in BA concentration. The determined reference limits generally exhibited better precision for haematological parameters, being lower than the upper recommended value 0.2, unlike biochemical parameters. Haematological parameters showed notable differences between pregnant and non-pregnant women, while many biochemical parameters' RIs remained similar. Only ALT and GGT showed lower non-comparable RI upper limits in the population pregnant women. CONCLUSIONS: Spectrophotometric BA method showed satisfactory performance and all examined parameters were within the set criteria. Moreover, RIs for key biochemical and haematological parameters, including BAs, have been established for the first time in the population of Croatian pregnant women.

Targeted Mass Spectrometry-Based Assays for Relative Quantification of 30 Brain-Related Proteins and Their Clinical Applications.

Cerebrospinal fluid (CSF) is a promising clinical sample for identification of novel biomarkers for various neurological disorders. Considering its direct contact with brain tissue, CSF represents a valuable source of brain-related and brain-specific proteins. Multiple sclerosis is an inflammatory, demyelinating neurological disease affecting the central nervous system, and so far there are no diagnostic or prognostic disease specific biomarkers available in the clinic. The primary aim of the present study was to develop a targeted mass spectrometry assay for simultaneous quantification of 30 brain-related proteins in CSF and subsequently to demonstrate assay feasibility in neurological samples derived from multiple sclerosis patients. Our multiplex selected reaction monitoring assay had wide dynamic range (median fold range across peptides = 8.16 × 103) and high assay reproducibility (median across peptides CV = 4%). Candidate biomarkers were quantified in CSF samples from neurologically healthy individuals (n = 9) and patients diagnosed with clinically isolated syndrome (n = 29) or early multiple sclerosis (n = 15).

Visual assessment of hemolysis affects patient safety.

BACKGROUND: Manual handling of hemolyzed samples is not standardized and is vulnerable to errors. This study aimed to evaluate laboratory errors due to manual handling of hemolyzed samples and to assess the risk they might have for patient safety. METHODS: Data were retrospectively obtained from a laboratory information system for 25 emergency tests from hemolyzed samples. Hemolysis (concentration of free hemoglobin >0.5 g/L) was visually assessed by comparison with a color chart. The reference person reestimated the routinely assessed degree of hemolysis to all samples (n=3185) received in the laboratory in a 1-week period. For each test, the correct and incorrect way of handling results was determined. Risk assessment was performed according to ISO 14971 standard with five categories of risk (S1-S5) and error occurrence (O1-O5). RESULTS: In the studied period, the emergency laboratory received 495 hemolyzed samples (15.5%) with a total of 2518 laboratory test requests (15.5%): 102 (20.6%) of the reports from hemolyzed samples had a comment on hemolysis; 31% of the test results were handled incorrectly (20.7% due to the incorrect release of the test result despite hemolysis interference and 10.3% due to unnecessary suppression), accounting for 4.8% of the total test volume. Tests with the highest combination of risk and occurrence rate were troponin T, potassium and total bilirubin. CONCLUSIONS: Manual handling of hemolyzed samples may lead to risk of errors in reporting results for troponin T, potassium and total bilirubin, which may have an effect on clinical decision. In addition, unnecessary suppression of the sample results unaffected by hemolysis could affect patient outcome.

Various glycolysis inhibitor-containing tubes for glucose measurement cannot be used interchangeably due to clinically unacceptable biases between them.

BACKGROUND: The aim of our study was to determine the difference between glucose concentration measured 30 min after venipuncture in ice-chilled heparin plasma sample and all currently available citrate buffer-containing tubes (Greiner Glucomedics, Greiner FC Mix and Sarstedt GlucoEXACT) and still widely used sodium fluoride/potassium oxalate (NaF/Kox) tubes from Greiner. METHODS: Blood was collected from 20 healthy volunteers and 20 patients with diabetes into LiH, NaF/KOx, Glucomedics, FC mix and GlucoEXACT tubes. Glucose was measured within 30 min from blood sampling in duplicate on the Architect c8000 analyzer. Mean biases between all tube types were calculated and compared to the recommended criteria (1.95%). Additionally, glucose concentrations measured in all five tube types were compared using the Friedman test. RESULTS: In the entire studied population, glucose concentrations measured in Glucomedics, FC mix and GlucoEXACT were higher (7.3%, 3.2% and 2.0%, respectively) than in the ice-chilled LiH tubes. When all glycolysis inhibitor-containing tubes were compared, Glucomedics tubes significantly differed from GlucoEXACT and FC mix tubes (biases -4.9% and 4.0%, respectively). In addition, there was a significant difference between the NaF/KOx tube and Glucomedics, as well as FC mix tubes (biases 7.1% and 3.0%, respectively). CONCLUSIONS: Glucose concentrations measured in recommended ice-chilled lithium heparin- and citrate buffer-containing tubes are not comparable. Significant biases exist between various glycolysis inhibitor-containing tubes; therefore, they cannot be used interchangeably.

Key Performance Indicators to Measure Improvement After Implementation of Total Laboratory Automation Abbott Accelerator a3600.

The aim of the study was to estimate improvement of work efficiency in the laboratory after implementation of total laboratory automation (TLA) by Abbott Accelerator a3600 in the laboratory with measuring different key performance indicators (KPIs) before and after TLA implementation. The objective was also to recommend steps for defining KPIs in other laboratories. For evaluation of improvement 10 organizational and/or technical KPIs were defined for all phases of laboratory work and measured before (November 2013) and after (from 2015 to 2017) TLA implementation. Out of 10 defined KPIs, 9 were successfully measured and significantly improved. Waiting time for registration of samples in the LIS was significantly reduced from 16 (9-28) to 9 (6-16) minutes after TLA (P < 0.001). After TLA all tests were performed at core biochemistry analyzers which significantly reduced walking distance for sample management (for more than 800 m per worker) and number of tube touches (for almost 50%). Analyzers downtime and engagement time for analyzers maintenance was reduced for 50 h and 28 h per month, respectively. TLA eliminated manual dilution of samples with extreme results with sigma values increment from 3.4 to >6 after TLA. Although median turnaround time TAT for potassium and troponin was higher (for approximately 20 min), number of outliers with TAT >60 min expressed as sigma values were satisfying (>3). Implementation of the TLA improved the most of the processes in our laboratory with 9 out of 10 properly defined and measured KPIs. With proper planning and defining of KPIs, every laboratory could measure changes in daily workflow.

Releasing Dynamic of Serum ST2 and Calprotectin in Patients with Acute Ischemic Stroke.

This study investigated the releasing dynamics of serum ST2 and calprotectin in patients with acute IS. The study included acute IS patients (N = 20) with an NIH Stroke Scale score ≥8. Sampling was performed at seven time points: after admission (T0) and at the following 24 h consecutive intervals (T1-T6). Primary outcome at 90 days was evaluated using the modified Rankin scale: 0-2 for good and 3-6 for poor functional outcome. The secondary outcome was all-cause mortality after 90 days. Fifteen patients had a poor outcome, and eight died. Results showed a statistically significant difference in ST2 concentrations between good and poor outcomes at T0 (p = 0.04), T1 (p = 0.006), T2 (p = 0.01), T3 (p = 0.021), T4 (p = 0.007), T5 (p = 0.032), and for calprotectin T6 (p = 0.034). Prognostic accuracy was highest for ST2 at T1 for a cut-off > 18.9 µg/L (sensitivity 80% and specificity 100.0%) and for calprotectin at T5 for a cut-off > 4.5 mg/L (sensitivity 64.3% and specificity 100.0%). Serum ST2 and calprotectin-releasing dynamics showed a valuable prognostic accuracy for IS outcomes.

The Association of Serum Calprotectin with Fitness Indicators and Biochemical Markers in High-Level Athletes: A Continuous Dynamic Monitoring during One Competitive Season.

The objective was to determine the associations between several biochemical indicators and the dynamics of concentration change across four physical fitness phases over the period of a competitive season. Furthermore, associations between serum calprotectin and biomarkers of inflammation or muscle injury and physical indicators were examined. SUBJECTS AND METHODS: Twenty professional male water polo players (median age: 28 (22-42)) were included in this study. Serum creatine kinase activity was determined by the automated photometric UV method. The concentrations of calprotectin, C-reactive protein, and myoglobin were measured using an automated immunoturbidimetric method, while an automated immunochemistry method was employed for interleukin-6, troponin I, and cortisol determination. Tests of repeated strength, maximal strength, and static strength were used to evaluate physical activity. RESULTS: Serum calprotectin concentrations expressed in median and IQR were significantly different: T1: 2.92 g/mL (2.47; 3.86); T2: 2.35 g/mL (1.26; 2.87); T3: 2.27 g/mL (1.60; 3.27); and T4: 1.47 g/mL (1.04; 2.85) (p = 0.004). Cortisol concentration and CK activity showed significant changes among phases (p = 0.049 and p = 0.014, respectively). Each physical activity examined showed a significant seasonal decrease (all p values were 0.001). Calprotectin serum concentration and indicators of muscular injury, inflammation, and physical activity were found to be correlated during particular stages of the seasonal examination. CONCLUSIONS: Calprotectin values determined throughout one competitive season decreased as training intensity among water polo players increased. Serum calprotectin concentrations and indicators were related to biochemical markers of inflammation and muscle damage.

Dark brown serum and plasma samples: a case report.

This case report describes occurrence of unusual, dark brown coloration of citrate plasma and serum samples in a female 68 years old patient admitted into Emergency department (ED). Patient complained of nausea and vomiting, fever up to 38.9°C, colicky pain in abdomen, diminished urinary output and yellowish skin tone. Her medical history included arterial hypertension, hypothyroidism and facial squamous cell carcinoma. For previous two years, she was treated with tuberculostatic therapy for Mycobacterium avium positive interstitial lung disease. Regular follow-up showed no signs of active disease. Upon admission to ED, complete blood count (CBC) analysis showed low red blood count (RBC) (3.76 x1012/L (reference interval (RI) 3.86 - 5.08 x1012/L)), low haemoglobin (Hb) concentration (111 g/L (RI 119 - 157 g/L)) and low haematocrit (Hct) (0.310 L/L (RI 0.360 - 0.470 L/L)). Biochemistry analytes were high, with foremost lactate dehydrogenase (LD) activity (2900 U/L, RI < 240 U/L). After communication with the clinician, methaemoglobin measured in arterial blood gas sample was reported. Patient was admitted to the Intensive care unit and upon reflex testing of haptoglobin, intravascular haemolysis was confirmed. This case indicates that every case of brown coloration of the serum must be promptly communicated to the clinician. Reflex testing assured timely diagnosis and favourable patient outcome.

Third-party verification of total and specific immunoglobulin E on analyzer Immulite® 2000XPi.

BACKGROUND: Although Immulite®2000XPi (Siemens) allergy testing is routinely used in laboratories worldwide; a well-designed comprehensive third-party verification of its analytical performance has not yet been published. Our aim was to verify stability, precision and reference intervals for total and some frequent specific IgEs, test trueness for several most common allergens and assess the comparability of fluorescent- and chemiluminescent-enzyme immunoassay for total IgE. METHODS: Verification was based on EP15-A3, EP9-A3 and EP28-A3c CLSI guidelines. Stability was tested on serum pools stored at 2-8 °C, during 7 days. Manufacturer's reference intervals were verified for children and adults. Acceptance criteria were based on manufacturer specifications and national External Quality Assessment provider data. RESULTS: Samples for total IgE analysis were stable at 2-8 °C for 7 days. Precision was within manufacturer's specifications for most of tested allergens, except for total IgE (92.5 kIU/L, CVs 5.6% and 7.6%) and w1 (9.3 kU/L, CV = 5.9%). Bland-Altman plot revealed statistically significant constant (-32.4 kIU/L) and proportional bias (-20.2%) for total IgE between two methods. Passing Bablok showed statistically but not clinically significant biases between two Immulite analyzers for betula verrucosa (constant and proportional), house dust mite and egg white allergens (proportional). For adults and children, total IgE results were within reference intervals declared by Siemens. CONCLUSIONS: Immulite®2000XPi assay has acceptable precision and trueness. Samples for total IgE are stable even longer than declared by the manufacturer. Declared reference intervals can safely be adopted for both adult and children population in our laboratory. Fluorescent- and chemiluminescent-enzyme immunoassay results of total IgE are not comparable.

Haemolysis and lipemia interfere with resistin and myeloperoxidase BioVendor ELISA assays.

INTRODUCTION: The aim of our study was to investigate the influence of haemolysis and lipemia on resistin (RES) and myeloperoxidase (MPO) measurement by BioVendor enzyme-linked immunosorbent assays (ELISA). MATERIALS AND METHODS: Blood was taken from healthy volunteers into lithium heparin tubes. Plasma samples were spiked with Lipofundin® emulsion (B. Braun Melsungen AG, Germany) for lipemia interference testing. Haemolysed samples were obtained by drawing aliquots of heparinized blood through a 26 gauge needle. Index of haemolysis (H), lipemia (L) and triglyceride concentration were measured on Abbott Architect c8000. Haemoglobin concentration was measured on Sysmex XN-1000. Concentrations of RES and MPO in all samples were determined with RES and MPO ELISA kits (BioVendor, Czech Republic). All measurements were performed in triplicate. Biases from the native samples were calculated for both analytes and compared with an arbitrary value (e.g. ± 10%). RESULTS: Triglyceride concentration in the investigated samples ranged from 0.57 to 38.23 mmol/L, which corresponds to L index from - 0.01 to 13.77. Haemoglobin concentration in all samples ranged from 0 to 8 g/L which correspond to H index from 0.05 to 8.77. Both MPO and RES showed significant biases at 1 g/L haemoglobin (58.7% and 66.7%, respectively). Also, both MPO and RES showed significant biases at 4.66 mmol/L triglycerides (33.8% and - 12.2%, respectively). CONCLUSIONS: Resistin BioVendor assays are affected by haemolysis and lipemia already at low degree of interferent. Haemolysis was found to interfere at 1 g/L haemoglobin for both assays, while lipemia interferes at 4.66 mmol/L of triglycerides.

Clinician's opinion about critical risk results proposed by the Croatian Chamber of Medical Biochemists: a survey in one Croatian tertiary hospital.

INTRODUCTION: It has been recommended that each laboratory modify their critical result reporting practices to reflect the clinical needs of their patient populations. The aim of this survey was to assess how well critical laboratory values defined by the Croatian Chamber of Medical Biochemists (CCMB) correspond to the needs of the physicians at University hospital "Sveti Duh" (Zagreb, Croatia). MATERIALS AND METHODS: We conducted a survey among physicians from five departments in our hospital. Physicians were asked general questions about critical risk results (if and how they want to be informed). A list of critical risk results defined by the CCMB was offered and physicians were asked to revise the existing critical risk results and suggest adding new parameters. Obtained data were presented as numbers. Where the number of observations was low, ratios were used. RESULTS: Survey response rate was 43% (52/121). Majority (48/52) wants to be informed of critical risk results, either personally (31/48) or through a colleague (32/48). They prefer to be informed about critical risk results of prothrombin time, platelet count, haemoglobin, glucose, creatinine, sodium and potassium. Revisions in the CCMB critical risk result list are proposed by 13 out of 48 physicians. Neonatologists approved the CCMB's list. CONCLUSIONS: Although most critical risk results defined by the CCMB correspond well to the needs of the physicians in our hospital, some revisions are necessary to meet the particular needs of individual departments. Communication of critical risk results to those who have requested laboratory testing is highly appreciated practice.

Analytical verification of 12 most commonly used urine dipsticks in Croatia: comparability, repeatability and accuracy.

INTRODUCTION: Variability among manufacturers of urine dipsticks, respective to their accuracy and measurement range, may lead to diagnostic errors and thus create a serious risk for the patient. Our aims were to determine the level of agreement between 12 most commonly used urine dipsticks in Croatia, examine their accuracy for glucose and total protein and to test their repeatability. MATERIALS AND METHODS: A total of 75 urine samples were used to examine comparability and accuracy of 12 dipstick brands (Combur 10 TestM, ChoiceLine 10, Combur 10 TestUX, ComboStik 10M, ComboStik 11M, CombiScreen 11SYS, CombiScreen 10SL, Combina 13, Combina 11S, Combina 10M, UriGnost 11, Multistix 10SG). Agreement between each dipstick and the reference (Combur 10 TestM) was expressed as kappa coefficient (acceptable κ ≥ 0.80). Accuracy for glucose and total protein was tested by comparison with quantitative measurements on analysers: AU400 (Beckman Coulter, USA), Cobas 6000 c501 (Roche Diagnostics, Germany) and Architect plus c4000 (Abbott, USA). Repeatability was assessed on 20 replicates (acceptable > 90%). RESULTS: Best agreement was achieved for glucose, total protein and nitrite (11/11, k > 0.80) and the lowest for bilirubin (5/5, k < 0.60). Sensitivities for total protein were 41-75% (AU400) and 56-92% (Cobas and Architect); while specificities were 41-75% (AU400, Cobas, Architect). Dipsticks' sensitivity and specificity for glucose were 68-98%. Most of the dipsticks showed unacceptable repeatability (6/12, < 90%) for one parameter, most prominently for pH (3/12, < 90%). CONCLUSIONS: Most commonly used dipsticks in Croatia showed low level of agreement between each other. Moreover, their repeatability varies among manufacturers and their accuracy for glucose and proteins is poor.

Reporting categories in urine test strip analysis: Croatian survey and call for action.

INTRODUCTION: In line with the national recommendations, Croatian medical laboratories report urine test strip qualitative analysis results using a categorized scale with defined number of categories. Since concentration ranges for measured analytes have not been provided by national professional authority, it is up to the laboratories to define their own categories. The aim of study was to assess the comparability of concentrations assigned to different categories used in reporting the results of dipstick urinalysis in Croatian laboratories. MATERIAL AND METHODS: A questionnaire was e-mailed to all Croatian medical laboratories (N = 195). They were asked to provide the number of categories and respective concentrations for each parameter. Data were described as numbers and percentages. Values above the upper reference range limit, which were assigned as normal and/or trace category, were considered as false negative. RESULTS: Response rate was 71% (139/195). Seventy percent (98/139) of laboratories report their results with either higher (77/98; 79%) or lower (2/98; 2%) number of categories, relative to the national recommendation, whereas 19/98 (19%) report their results as concentrations. Great heterogeneity of reporting categories was observed. Multiple categories were assigned to same concentrations and there was a large overlap of concentrations for most categories. Considerable proportion of laboratories reported false negative results for ketones (42%), leukocytes (30%) and glucose (21%). CONCLUSIONS: The concentrations assigned to categories used to report the results of dipstick urinalysis are not comparable among Croatian medical laboratories. There is an urgent need for harmonization and standardization of reporting the results of urine dipstick analysis in Croatia.

The knowledge and understanding of preanalytical phase among biomedicine students at the University of Zagreb.

INTRODUCTION: The educational program for health care personnel is important for reducing preanalytical errors and improving quality of laboratory test results. The aim of our study was to assess the level of knowledge on preanalytical phase in population of biomedicine students through a cross-sectional survey. MATERIALS AND METHODS: A survey was sent to students on penultimate and final year of Faculty of Pharmacy and Biochemistry--study of medical biochemistry (FPB), Faculty of Veterinary Medicine (FVM) and School of Medicine (SM), University of Zagreb, Croatia, using the web tool SurveyMonkey. Survey was composed of demographics and 14 statements regarding the preanalytical phase of laboratory testing. Comparison of frequencies and proportions of correct answers was done with Fisher's exact test and test of comparison of proportions, respectively. RESULTS: Study included 135 participants, median age 24 (23-40) years. Students from FPB had higher proportion of correct answers (86%) compared to students from other biomedical faculties 62%, P < 0.001. Students from FPB were more conscious of the importance of specimen mixing (P = 0.027), prevalence of preanalytical errors (P = 0.001), impact of hemolysis (P = 0.032) and lipemia interferences (P = 0.010), proper choice of anticoagulants (P = 0.001), transport conditions for ammonia sample (P < 0.001) and order of draw during blood specimen collection (P < 0.001), in comparison with students from SM and FVM. CONCLUSIONS: Students from FPB are more conscious of the importance of preanalytical phase of testing in comparison with their colleagues from other biomedical faculties. No difference in knowledge between penultimate and final year of the same faculty was found.

Blood gas testing and related measurements: National recommendations on behalf of the Croatian Society of Medical Biochemistry and Laboratory Medicine.

Blood gas analysis (BGA) is exposed to risks of errors caused by improper sampling, transport and storage conditions. The Clinical and Laboratory Standards Institute (CLSI) generated documents with recommendations for avoidance of potential errors caused by sample mishandling. Two main documents related to BGA issued by the CLSI are GP43-A4 (former H11-A4) Procedures for the collection of arterial blood specimens; approved standard - fourth edition, and C46-A2 Blood gas and pH analysis and related measurements; approved guideline - second edition. Practices related to processing of blood gas samples are not standardized in the Republic of Croatia. Each institution has its own protocol for ordering, collection and analysis of blood gases. Although many laboratories use state of the art analyzers, still many preanalytical procedures remain unchanged. The objective of the Croatian Society of Medical Biochemistry and Laboratory Medicine (CSMBLM) is to standardize the procedures for BGA based on CLSI recommendations. The Working Group for Blood Gas Testing as part of the Committee for the Scientific Professional Development of the CSMBLM prepared a set of recommended protocols for sampling, transport, storage and processing of blood gas samples based on relevant CLSI documents, relevant literature search and on the results of Croatian survey study on practices and policies in acid-base testing. Recommendations are intended for laboratory professionals and all healthcare workers involved in blood gas processing.

Interference of M-protein on prothrombin time test - case report.

The aim of this report was to present a case of interference on prothrombin time (PT) test that directed further laboratory diagnostics and resulted with final detection of monoclonal gammopathy in an 88-year old man. Routine coagulation testing during medical examination at Emergency Department revealed unmeasurable PT (< 7% activity) and activated partial thromboplastin time (aPTT) within reference range. After repeated sampling for coagulation testing, PT was unmeasurable again, as well as fibrinogen level (< 0.8 g/L), thrombin time (TT) was significantly prolonged (107 seconds) and aPTT was within reference range. In both plasma samples refrigerated at 4 ˚C overnight, white gelatinous precipitate was visible between the cell and plasma layers and the presence of monoclonal protein (M-protein) was suggested in our patient. Further laboratory diagnostics revealed total serum proteins at concentration of 123 g/L and the presence of M-protein IgG lambda (λ) at concentration of 47.1 g/L. These results suggested monoclonal gammopathy as an underlying pathophysiological condition in our patient. Activities of coagulation factors II, V, VII and X were within reference ranges or increased. These results and correction of unmeasurable PT result to 67% in mixing test with commercial normal plasma suggest in vitro rather than in vivo interference of M-protein on PT result. In contrast, significantly prolonged TT results in all analysed samples suggest impact of M-protein on this global coagulation test due to possible effect on fibrin polymerization.

The role of human kallikrein 6, clusterin and adiponectin as potential blood biomarkers of dementia.

OBJECTIVES: Progressive degenerative syndromes which affect brain, altering memory, behavior, cognition and emotion, are commonly defined as dementia. It was suggested that serum human kallikrein 6 (KLK6), clusterin (CLU) and adiponectin (ADPN) in combination with inflammation markers, neuroimaging and neuropsychological testing could assist in discriminating dementia patients from control individuals. Our aim was therefore to compare serum concentrations of KLK6, CLU and ADPN and inflammatory marker, interleukin-6 (IL-6), in patients suffering from Alzheimer's disease (AD), patients with vascular dementia (VAD), cognitively healthy participants (CHP) and those with mild cognitive impairment (MCI). DESIGN AND METHODS: Serum samples were collected from AD, VAD and MCI patients admitted to the University Department of Neurology (Zagreb, Croatia) for regular follow-up. All patients underwent standard neuroimaging procedures including brain CT, neurosonological assessment with intima-media thickness (IMT) and breath holding index (BHI) calculations. Cognitive abilities were tested using standard Mini-Mental State Examination (MMSE) and Montreal Cognitive Assessment (MoCA). Concentrations of KLK6, CLU, ADPN and IL-6 were determined in all serum samples. RESULTS: We have recruited a total of 235 participants, divided in 4 groups: AD (N=70), VAD (N=67), MCI (N=48) and CHP (N=50). Serum concentrations of KLK6 (P=0.137), CLU (P=0.178) and ADPN (P=0.268) did not differ between AD, VAD, MCI and cognitively healthy control group of participants, whereas IL-6 was significantly higher in VAD patients than in AD, MCI and CHP individuals (P=0.014). There was no association between investigated biomarkers and clinical patient parameters. CONCLUSIONS: Serum concentrations of KLK6, CLU and ADPN do not differ between AD, VAD and controls with and without mild cognitive impairment. Higher IL-6 levels in VAD group point to the inflammatory component in the development of vascular dementia. Investigated biomarkers are not associated with neuroimaging findings and neuropsychological patient data.

Short-term and long-term storage stability of heparin plasma ammonia.

AIMS: Ammonia is an extremely unstable analyte and requires special attention during sampling, transport and storage. The aim of this study was to evaluate the stability of ammonia in lithium-heparin plasma during short-term (at +4°C) and long-term (at -20°C) storage. METHODS: Twenty plasma samples were used for short-term stability assessment. Each sample was divided into five aliquots and stored in stoppered tubes at +4°C, for 1, 2, 3, 4 and 24 h from initial testing. Fifteen plasma samples were used for long-term stability assessment. Each sample was divided into eight aliquots and stored in stoppered tubes at -20°C for 3, 24, 48 h and 1, 2, 4, 8 and 12 weeks from initial testing. Ammonia concentration was determined on a Beckman Coulter AU2700 chemistry analyser using Randox ammonia enzymatic UV method. Bias was calculated from initial value for each time point and compared with quality specifications defined by Royal College of Pathologists of Australasia. RESULTS: The average bias exceeded the total allowable error after storage of samples for 1 h at +4°C and 3 h at -20°C. CONCLUSION: Ammonia is not stable during storage at +4°C and -20°C in lithium-heparinised plasma and should therefore be analysed immediately.