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Poster Abstracts – IFCC WorldLab 2017 – Durban, South Africa, 22-25 October 2017 • DOI 10.1515/cclm-2017-7062
Clin Chem Lab Med 2017; 55, Special Suppl, pp S1483 – S1505, October 2017 • Copyright © by Walter de Gruyter • Berlin • Boston
T027
Metabolomics and biomarkers
NEW APPROACH FOR DIAGNOSTICATION OF METABOLIC SYNDROME
G. Atanasova 2, P. Yordanova - Laleva 4, M. Marinov 1, M. Atanasov 3
1
Dean, Aviation Faculty, Vasil Levski National Military University, Bulgaria.
2
Department of Internal Medicine, Pleven Medical University, Bulgaria
3
Head of Department “Organization and management of tactical units and aviation armament”, Aviation faculty, National
Military University, Bulgaria.
4
Pleven Medical University, Bulgaria
BACKGROUND-AIM
The metabolic syndrome is a series of synergistically interacting risk factors for CVD, many or all of which may share a
common etiology, at least in a substantial proportion of patients. Delineation of the including heterogeneity of etiology
would be useful in refining prevention and treatment strategy.
Objectives of this study is to evaluate opportunities of using of mean arterial pressure (MAP) as a component of the
metabolic syndrome (MS) instead systolic and diastolic blood pressures (SBP and DBP) and to evaluate influence of the
MS and its components on pulse pressure (PP) and apolipoprotein B/Apolipoprotein A1 (Apo B/Apo A1).
METHODS
A total of 104 persons without any apparent disease were selected. Among these people MS was found in 35, according
to NCEP-ATP III definition. One way ANOVA test, multiple comparison tests of means and multiple logistic regression
analyses were used. The MAP was obtained by the formula MAP=SBP/3+2*DBP/3. The four groups used in ANOVA are
men and women with and without MS. The ANOVA F-statisticis 3.683 with p-value 0.0145. The first logistic regression
includes gender, PP and Apo B/Apo A1.
RESULTS
The average value of mean arterial pressure for all personswas 95.17 [mm Hg] and the standard deviation was 10.65
[mm Hg]. Pulse pressure and Apo B/Apo A1 ratio could be used as complex marker for MS.
The ANOVA F-statistic is 17.71 with p-value less than 0.00001. The obtained model showed that for increase of PP with
5 mm Hg it was expected about 1.2314 times increase in the odds ratio of MS and for increase of APO B/APO A1 ratio
with 0.1 it was expected about 1.6363 times increase in the odds ratio.
CONCLUSIONS
Reducing the number of used biochemical marker could improve the cost efficiency in the diagnostication of MS. MAP
showed itself as a promising indicator, which after some broader studies could replace SBP and DBP in the MS
definition.
When the pulse pressure was wide and wriest was grater than 102/88 cm (men/women) the odds ratio was above 1.
These two factors could be used to diagnose metabolic syndrome. The same conclusion could be made for wide pulse
pressure and triglycerides level grater than 1.7 mmol/l. The results showed that PP and wriest or triglycerides level could
be used as indicator of metabolic syndrome.
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S1483
Poster Abstracts – IFCC WorldLab 2017 – Durban, South Africa, 22-25 October 2017 • DOI 10.1515/cclm-2017-7062
Clin Chem Lab Med 2017; 55, Special Suppl, pp S1483 – S1505, October 2017 • Copyright © by Walter de Gruyter • Berlin • Boston
T028
Metabolomics and biomarkers
RESISTIN IS A MARKER OF DISEASE ACTIVITY IN PATIENTS WITH INFLAMMATORY BOWEL DISEASE
J. Berska 1, J. Bugajska 1, M. Zwolińska-Wcisło 2, K. Sztefko 1
1
Department of Clinical Biochemistry, Institute of Pediatric, Jagiellonian University Medical College, Krakow, Poland
2
Gastroenterology and Hepatology Clinic Jagiellonian University Medical College, Krakow, Poland
BACKGROUND-AIM
Epidemiological data suggest that incidence of inflammatory bowel diseases (IBD) such as ulcerative colitis (UC) and
Crohn’s disease (CD) is increasing in different regions around the world. The disease is characterized by recurring
episodes of inflammation of the mucosal layer of the large bowel. Many evidences suggest that adipokines play an
important role in mucosal inflammation of the gastrointestinal tract and might serve as markers of inflammatory activity.
The aim of the study: to assess the relationship between the serum resistin level and the disease activity in patients with
inflammatory bowel disease.
METHODS
The study included 57 patients with UC or CD (M/F 29/28, mean age 42.1±16.2 years). The clinical activity of the disease
was assessed using The Mayo Clinic Disease Activity Index (Mayo) and the Crohn’s Disease Activity Index (CDAI).
Among UC patients 13 had active and 13 had non-active form of the disease. Among CD patients 19 had active and 12
had non-active form of the disease. Resistin concentration was measured in serum using enzyme immunoassay (ELISA,
RD Systems). For statistical evaluation of the results mean±SE, Student’s t test, Pearson’s correlation coefficient and
One-Way ANOVA with Tukey post-hock test were used. A level of p<0.05 was considered statistically significant. The
ROC curve (Receiver Operating Characteristic) for resistin was constructed and the AUC (the Area under the Curve) was
computed.
RESULTS
The mean value of serum resistin level was significantly higher in patients with active than in those in non-active IBD
(19.6±2.63 ng/ml vs 10.6±1.9 ng/ml; p<0.007). Patients with UC as well as with CD had the mean value of resistin
concentration higher in active than in non-active form of the disease, but the difference was statistically significant only
for patients with UC (p<0.02). Positive trend between resistin level and clinical activity index of the ulcerative colitis has
been observed but the difference did not reach statistical significance (r=0.382, p=0.06). ROC curve for the
discrimination active from non-active form of the disease show the AUC=0.716, and cut-off point 16.2 ng/ml.
CONCLUSIONS
Resistin levels may be used as independent marker of disease activity in patients with inflammatory bowel disease.
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S1484
Poster Abstracts – IFCC WorldLab 2017 – Durban, South Africa, 22-25 October 2017 • DOI 10.1515/cclm-2017-7062
Clin Chem Lab Med 2017; 55, Special Suppl, pp S1483 – S1505, October 2017 • Copyright © by Walter de Gruyter • Berlin • Boston
T029
Metabolomics and biomarkers
PARAMETERS OF IRON STATUS IN DIFFERENTIAL DIAGNOSIS OF ANEMIA
D. Ćujić 3, M. Savković 1, S. Simić-Ogrizović 2, S. Ignjatović 1, V. Dopsaj 1
1
Center for Medical Biochemistry, Clinical Center of Serbia, Belgrade, Serbia;Faculty of Pharmacy, University of
Belgrade, Belgrade, Serbia
2
Clinic of Nephrology, Clinical Center of Serbia, Belgrade, Serbia; School of Medicine, University of Belgrade, Serbia
3
Institute for The Application of Nuclear Energy INEP, University of Belgrade, Belgrade, Serbia
BACKGROUND-AIM
Different etiological factors may cause disturbances in iron metabolism, leading to anemia due to iron deficiency. Iron
metabolism is under direct control of hepcidin-25 (Hep), a peptide hormone synthesized in the liver. Hence,
measurement of Hep could be a valuable parameter in assessment of iron status, differential diagnosis of anemia or
monitoring treatment in anemic patients.
The aim of this study was to explore Hep levels in different types of anemia related to iron metabolism disturbances, as
well as Hep relation to other parameters of iron status.
METHODS
Hemodialysis-dependent chronic kidney diseases patients (HD group, n=111) and patients with iron deficiency anemia
(IDA group, n=19) were included in the study. Concentrations of biochemical parameters serum iron (Fe), transferin (TR),
ferritin (FER) and Hep were determined using commercial diagnostic tests, while hematological parameters were
determined on hematology analyzer. Descriptive statistics and correlation analysis were used for data analysis.
RESULTS
The mean Fe in HD and IDA groups were 12.9 mol/L and 6.3 mol/L respectively. In IDA group, low levels of both Hep
(3.0 ng/mL) and FER (8.9 g/L) were measured, while in HD group were elevated, 74.5 ng/mL for Hep and 456,6 ng/mL
for FER. Positive correlation between Hep and FER was seen in both groups, r = 0.7788 (p<0.05) for IDA and r = 0.7789
(p<0.05) for HD. Mean TR in IDA patients was 3,2 g/L, while low TR was measured in HD group, 1,7 g/L. Negative
correlation was observed between Fe and TR in IDA patients (r = -0.6483, p<0.05). In both study groups, hemoglobin
levels were low, 105 g/L in HD and 102 g/L in IDA. Patients with IDA had lower MCV values (77 fL in IDA compared to 93
fL in HD), as well as reticulocyte % (1.4 % in IDA versus 2.0 % in HD). In patients with HD, Hep levels were in negative
correlation with hemoglobin (r = -0.3568, p<0.05), as well as with erythrocyte number (r = -0.3384, p<0.05).
CONCLUSIONS
Although Hep is assumed to be a major regulator of iron metabolism, the correlation between serum iron and Hep is not
necessarily seen in patients with iron metabolism disturbances. Multiple parameters are neccesery in order to assess
iron status in patients with anemia caused by low iron content.
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S1485
Poster Abstracts – IFCC WorldLab 2017 – Durban, South Africa, 22-25 October 2017 • DOI 10.1515/cclm-2017-7062
Clin Chem Lab Med 2017; 55, Special Suppl, pp S1483 – S1505, October 2017 • Copyright © by Walter de Gruyter • Berlin • Boston
T030
Metabolomics and biomarkers
SALIVARY BIOMARKERS IN PATIENTS WITH TEMPOROMANDIBULAR DISORDERS
J. Djordjevic 1, A. Miletic 2, D. Matanovic 3
1
Center for Medical Biochemistry, Clinical Center Serbia, Belgrade
2
Clinic for Prosthodontics, School of Dental Medicine, University of Belgrade, Belgrade, Serbia
3
Clinic for Rehabilitation, Faculty of Medicine, University of Belgrade, Belgrade, Serbia
BACKGROUND-AIM
Temporomandibular disorders (TMDs) represent a group of musculoskeletal disorders affecting temporomandibular
joints (TMJs) and masticatory muscles, including other associated structures.After the odontogenic pain, TMDs are the
second cause of pain in the orofacial region.
This study examines salivary alpha-amylase and salivary cortisol levels which have been suggested as markers for
sypathetic nervous syistem (SNS) and HPA activity, respectively.
METHODS
At the School of Dental Medicine, Belgrade, a total of 18 TMD patients and 22 healthy adults were selected to participate
in this study. The subjects were evaluated using the Research Diagnostic Criteria for TMD (RDC/TMD). Statistical
analysis were performed using the SPSS®21 software. Mann-Whitney U test was used to compare salivary
alpha-amylase levels and T-test was used to compare salivary cortisol levels between groups.
RESULTS
Statisticaly significant difference between morning salivary alpha-amylase levels in TMD group (Median=33425U/L;
min=807 U/L, max=257500U/L) and the control group (Median=8950 U/L, min=720U/L, max=133232U/L (p=0.017) was
recorded. No statistically significant difference was observed in salivary cortisol levels (Mean±SD(TMD)=16.7±10.1
nmol/l, Mean±SD(control)=18.2±6.4 nmol/l; p=0.58).
CONCLUSIONS
Present findings showed higher salivary alpha-amylase levels in TMD patients compared to control group, while no
difference was recorded in salivary cortisol levels. Results of this study could indicate the relation between altered
symathetic nervous system activity and temporomandibular disorders. However, the relation between
temporomandibular disorders, pain, and sypathetic nervous syistem activity is complex and warrants further investigation
on a larger sample.
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S1486
Poster Abstracts – IFCC WorldLab 2017 – Durban, South Africa, 22-25 October 2017 • DOI 10.1515/cclm-2017-7062
Clin Chem Lab Med 2017; 55, Special Suppl, pp S1483 – S1505, October 2017 • Copyright © by Walter de Gruyter • Berlin • Boston
T031
Metabolomics and biomarkers
INVESTIGATING SERUM LEPTIN AND GHRELIN LEVELS AS METABOLIC SYNDROME BIOMARKERS IN ADULT
BLACK ZAMBIANS WITH TYPE 2 DIABETES MELLITUS AT THE UNIVERSITY TEACHING HOSPITAL, LUSAKA,
ZAMBIA
C.E. Goma 3, T. Kaile 1, B. Kamanga 2
1
UNIVERSITY OF ZAMBIA, SCHOOL OF MEDICINE DEAN. LUSAKA
2
University Teaching Hospital, Dept of internal medicine and endocrinology, Lusaka, Zambia.
3
University Teaching Hospital, Dept of Pathology and Microbiology, Lusaka, Zambia.
BACKGROUND-AIM
Metabolic Syndrome (MetS) is the clustering of at least three of the five medical conditions: elevated blood pressure,
central obesity, high fasting serum triglycerides, elevated fasting plasma glucose and low high-density lipoprotein levels.
Obesity is an established high risk factor for Type 2 diabetes mellitus (T2DM) and is a central component of the MetS.
The prevalence of MetS is on the increase, worldwide. Some of the biomarkers for MetS include Leptin and Ghrelin
levels. Leptin works through the leptin receptor (LEPR) also known as the obese receptor (OBR). Ghrelin activates also
Growth hormone-releasing hormone (GH-RH) neurons in the hypothalamic arcuate nucleus. Leptin is a peptide produced
by differentiated adipocytes. Leptin is a key hormone in the regulation of body fat stores. This peptide controls energy
metabolism at the level of hypothalamus by supporting the food intake and stimulating energy expenditure. Ghrelin is
able to modify glucose and insulin metabolism, blood adipogenesis and inflammatory processes in experimental
conditions.
The aim of this study was to determine the levels of Ghrelin and Leptin in adult Zambians with T2DM.
METHODS
The study was an Analytical Cross- Sectional hospital based research at medical out-patient clinic at the University
Teaching Hospital. This study screened 512 patients that were tested to determine the prevalence of MetS in Zambia
and 158 patients with T2DM and T2DM/ MetS were selected for the study, 79 adult black Zambian patients aged 18
years and above with T2DM and MetS were followed up for the findings. A control group comprised 63 diabetic patients
without MetS matched by age and sex. Serum fasting Leptin and Ghrelin fasting were measured using the Enzyme
Linked Immunosorbent Assay method.
Data was performed by using STATA statistics version 21 for windows and Microsoft excel 2010 for windows. To
compare means of the variables and determine any difference between metabolic syndrome and controls.
RESULTS
Type 2 diabetic patients with Metabolic Syndrome (MetS) (60%) were significantly (P=0.01) T2DM than those without
(39%). Plasma total triglyceride (TG) (215.9 ± 16.91) and blood pressure (148.8 ± 3.247) were strongly significantly high
(P<0.001) in patients with MetS as compared to those without MetS (104.4 ± 6.766 and 122.8 ± 2.725, respectively).
Fasting serum leptin levels was (57%) higher and fasting ghrelin was (76%) lower than normal.
CONCLUSIONS
Zambian Adult Metabolic Syndrome patients are at a greater risk of developing cardiovascular disorders. Interventions/
management should be put in place to help those patients avoid/delay onset of cardiovascular complications anticipated
upon accumulations of predisposing factors that are components of Metabolic Syndrome.
Key words: Metabolic Syndrome, Diabetes Mellitus, Leptin, Ghrelin
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S1487
Poster Abstracts – IFCC WorldLab 2017 – Durban, South Africa, 22-25 October 2017 • DOI 10.1515/cclm-2017-7062
Clin Chem Lab Med 2017; 55, Special Suppl, pp S1483 – S1505, October 2017 • Copyright © by Walter de Gruyter • Berlin • Boston
T032
Metabolomics and biomarkers
OPTIMIZATION OF M1 AFLATOXIN HPLC ANALYSIS METHOD IN COW AND MOTHERS’ MILK
R. Ghali 1, K. Diallo 1, E. Nouiri 1, S. Ouihibi 1, M. Araoud 1, A. Hedili 1
1
Research Laboratory of Environment and Toxicology LR12SP07, Tunisia
BACKGROUND-AIM
M1 aflatoxin Cow and mother Milks contamination is a serious public health concerns. The occurrence of this mycotoxin
is an important biomarker of B1 Aflatoxin exposure and it’s known as a potent carcinogenic and has been associated
with some impaired children growth.
METHODS
A validate Analytical method was established in our laboratory, Extraction and the purification of AFM1 from milk was
realized by immunoaafinty columns. Quantification was carried out by a Fluorescence-HPLC technique after a pre
column amplification of the natural toxin fluorescence by the TFA derivatisation.
RESULTS
The obtained results show an accurate, simple, specific and linear method in the range from 0.03 to 5 g mL-1 with an
excellent recovery. Detections and quantification limits were respectively, 0.01 and 0.03 g/L. This method is suitable for
AFLM1 quantification in milk. Analysis of naturally contaminated Tunisian Milk samples showed that 14% of the samples
are contaminated. Indeed, the most significant percentages of contamination are found in the cow milk samples with
levels ranging between 0.08 and 0.2 g/L. Although, commercial UHT cow milk had 20% incidence and the two women
analyzed samples showed no contamination by AFM1. In a total of 52 contaminated cow milk samples, 12.5% of them
contain AFLM1 at levels exceeding European legislated limit of 0.05 g/L.
CONCLUSIONS
Finding suggests the obligation to introduce AFLM1 analysis as a routine control of infant milk food based products, the
establishment of dietary recommen¬dations and the evaluation of breast milk AFM1 content, during pregnancy and
lactation.
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S1488
Poster Abstracts – IFCC WorldLab 2017 – Durban, South Africa, 22-25 October 2017 • DOI 10.1515/cclm-2017-7062
Clin Chem Lab Med 2017; 55, Special Suppl, pp S1483 – S1505, October 2017 • Copyright © by Walter de Gruyter • Berlin • Boston
T033
Metabolomics and biomarkers
INCREASED EXPRESSION OF GLUTAMINASE IN B LYMPHOCYTES FROM IMMUNE THROMBOCYTOPENIC
PURPURA (ITP) PATIENTS
M. Jiang 1, Y. Zhao 1, Y. He 1
1
Jiangsu Institute of Hematology, the First Affiliated Hospital of Soochow University, Suzhou
BACKGROUND-AIM
Autoreactive B lymphocytes continuing to produce antiplatelet antibodies is the underlying cause of immune
thrombocytopenic purpura (ITP), and subsequent phagocytosis of platelets by antibody-mediated macrophages is a
major direct cause of ITP. In this work, we intend to investigate glutamine metabolism in B lymphocytes from ITP
patients.
METHODS
Twelve acute ITP patients with PAIgG+, 8 ITP patients with PAIgG- after immunosuppressive therapy and 10 heath
people were recruited. Using FACS technique, B lymphocytes were isolated from peripheral blood. The expression of
glutaminase in B lymphocyte was examined using western blot and ELISA analysis. The secretion of PAIgG was
measured by MAIPA assay.
RESULTS
The expression and activity of glutaminase in B lymphocytes were increased in ITP patients compared with normal
individuals. This increase was more notable in PAIgG+ patients. And after treat with acivicin, an glutaminase inhibitor,
the PAIgG secretion of B lymphocyte was partly inhibited.
CONCLUSIONS
These results imply that in the pathogenesis of ITP, glutamine metabolism mediated by glutaminase plays an important
role in B lymphocyte function.
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S1489
Poster Abstracts – IFCC WorldLab 2017 – Durban, South Africa, 22-25 October 2017 • DOI 10.1515/cclm-2017-7062
Clin Chem Lab Med 2017; 55, Special Suppl, pp S1483 – S1505, October 2017 • Copyright © by Walter de Gruyter • Berlin • Boston
T034
Metabolomics and biomarkers
COMPARISON BETWEEN SERUM ACTIVE-B12 (HOLOTRANSCOBALAMIN) AND TOTAL VITAMIN B12 AS
MARKERS OF VITAMIN B12 STATUS
J. Jurkeviciene 2, L. Gogeliene 2, D. Vitkus 1
1
CENTER OF LABORATORY MEDICINE OF VILNIUS UNIVERSITY HOSPITAL SANTAROS KLINIKOS, VILNIUS
UNIVERSITY, FACULTY OF MEDICINE OF PHYSIOLOGY, BIOCHEMISTRY, MICROBIOLOGY AND LABORATORY
MEDICINE
2
LABORATORY OF BIOCHEMISTRY, CENTER OF LABORATORY MEDICINE OF VILNIUS UNIVERSITY HOSPITAL
SANTAROS KLINIKOS
BACKGROUND-AIM
Vitamin B12 (VB12) is bound to two carrier proteins – transcobalamin 1(TC1), metabolically inactive fraction, and
transcobalamin 2 (TC2), active fraction. It is generally recommend that patients with symptoms of VB12 deficiency
anemia and patients with suspected neuropsychiatric abnormalities should be tested for VB12 deficiency. A new test
which measures active VB12 fraction became available recently. The aim of the study was to evaluate the relationship
between total VB12 and active B12 concentration in serum.
METHODS
We analyzed 73 samples of patients referred to the laboratory for the assessment of VB12 status. A wide range of
clinical status were present in patients including anemia, gastrointestinal and endocrine diseases, malignancy and renal
failure. Serum levels of total VB12 and active VB12 were measured on ADVIA Centaur®XP analyzer (Siemens).
RESULTS
Results were divided into three groups according to the serum concentration level of total VB12: the group with possible
VB12 deficiency (<148 pmol/L), the “grey-zone” group with suspected VB12 deficiency (from 148 to 258 pmol/L), and the
group with VB12 deficiency unlikely (>258 pmol/L). SPSS software was used for statistical data analysis. Statistical
correlation between whole group of total VB12 and active-B12 show that there is a statistically significant correlation
between the two assays (p=0.000), positive Pearson correlation r=0.645, correlation based on Cohen’s
recommendations was strong. The Pearson correlation coefficient between the “grey-zone” total VB12 and active VB12
was r=0.262 and indicated no relationship between two variables.
CONCLUSIONS
Findings suggest that two assays agree well within the range of possible VB12 deficiency and when the lack of VB12 is
unlikely. It was shown disagreement in results of 6 samples in “grey-zone” of total VB12 where active VB12 was low, and
3 samples results in the group with VB12 deficiency unlikely, where active VB12 was low. It indicates that 9 patients of
73 could be potentially misleadingly classified as VB12 deficiency unlikely, if only total VB12 results have been used.
There is an evidence that active VB12 assay has better diagnostic accuracy than the total VB12 assay in patients with
results in “grey-zone”.
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S1490
Poster Abstracts – IFCC WorldLab 2017 – Durban, South Africa, 22-25 October 2017 • DOI 10.1515/cclm-2017-7062
Clin Chem Lab Med 2017; 55, Special Suppl, pp S1483 – S1505, October 2017 • Copyright © by Walter de Gruyter • Berlin • Boston
S1491
T035
Metabolomics and biomarkers
ASSOCIATION OF BRANCHED-CHAIN
CARDIO-METABOLIC RISK FACTORS
AMINO
ACIDS
AND
AROMATIC
AMINO
ACIDS
WITH
L. Khambule 2, S. Norris 2, N. Crowther 1, T. Snyman 1, J. George 1
1
National Health Laboratory Services
2
University of the Witwatersrand
BACKGROUND-AIM
A number of studies have shown that serum levels of branched chain amino acids (BCAAs: valine, leucine, isoleucine)
and aromatic amino acids (AAAs: tyrosine, phenylalanine) are elevated in a variety of cardio-metabolic diseases in
population groups resident in high income countries. In this study, we sought to describe the association of BCAAs and
AAAs with the metabolic syndrome and its individual components i.e. HDL-cholesterol, triglyceride, blood pressure and
plasma glucose concentrations in Black African (BAs) and Asian Indian (AI) populations in South Africa.
METHODS
We used serum samples collected from AI (n=349) and BA (n=369) subjects recruited through the Birth to Twenty Study
to measure blood levels of BCAAs and AAAs. Existing data on fasting blood glucose, lipid profile, blood pressure, waist
circumference and body mass index (BMI) as well as measurements of visceral and subcutaneous abdominal fat were
obtained for all individuals. The BCAAs and AAAs were quantified using liquid chromatography mass spectrometry
(LC-MS). Multivariable regression models were created to test for the association of amino acid levels (included as
independent variables) with metabolic syndrome and its components (included as dependent variables) with adjustment
for possible confounders. Furthermore, the determinants of amino acid levels were assessed using multivariable
regression models with the amino acids included as the dependent variable. Amino acid levels were compared across
tertiles of visceral fat thickness using ANOVA.
RESULTS
The serum total amino acid levels (sum of BCAAs and AAAs) were higher in AIs compared to BAs (p=0.004). The
BCAAs and AAAs were significantly and positively associated with metabolic syndrome and its individual components,
particularly in AIs, in unadjusted regression models but most of these associations were lost after adjustment for age,
gender, BMI, smoking, education, visceral fat and subcutaneous fat. The loss of significance was largely due to visceral
fat. However, triglyceride concentration was still significantly associated with total amino acids, valine and leucine levels
in BAs even after adjustment for the above co-variates (p<0.01 for all associations). In multivariable linear regression
models, visceral fat was the principal determinant of total amino acid levels in both BAs (p=0.002) and AIs (p<0.001).
Individuals with visceral fat thickness in the top tertile (≥ 6cm) had significantly higher total amino acid levels compared to
those with a visceral fat thickness in the bottom tertile (≤ 2.9cm) in BAs (p=0.012) and AIs (p<0.001).
CONCLUSIONS
Visceral fat, through an unknown mechanism, is positively related to total serum amino acid levels in AIs and BAs whilst
valine and leucine are strong indicators of hypertriglyceridaemia in BAs. Longitudinal studies are required to investigate
the ability of these amino acids to predict hypertriglyceridaemia in the BA population.
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Poster Abstracts – IFCC WorldLab 2017 – Durban, South Africa, 22-25 October 2017 • DOI 10.1515/cclm-2017-7062
Clin Chem Lab Med 2017; 55, Special Suppl, pp S1483 – S1505, October 2017 • Copyright © by Walter de Gruyter • Berlin • Boston
T036
Metabolomics and biomarkers
APOPTOSIS OF COLON CANCER CELLS UNDER THE EFFECT OF GELDANAMYCIN DERIVATE
F. Kosova 6, Z. Kasar 2, I. Tuglu 3, F. Ozdal Kurt 1, S. Gok 5, Z. Ari 4
1
Department of Biology, Celal Bayar University Science and Art Faculty, Manisa, Turkey
2
Department of Chemistry, Celal Bayar University Science and Art Faculty, Manisa, Turkey,
3
Department of Histology and Embryology, Celal Bayar University Medical Faculty, Manisa, Turkey
4
Department of Medical Biochemistry, Celal Bayar University Medical Faculty, Manisa, Turkey
5
Department of Pharmacology, Celal Bayar University Medical Faculty, Manisa, Turkey
6
Faculty of Health Science, Medical Biochemistry, Celal Bayar University, Manisa, Turkey
BACKGROUND-AIM
The apoptotic effect of geldanamycin derivative may be important for the colorectal cancer therapy. The
mechanisms of apoptosis require understanding of the behavior of colon cancer cell line Colo-205 which mimics colon
adenocarcinoma. Therefore, the effect of IC50 dose of 17-allylamino-17-demethoxygeldanamycin (17- AAG) on the colon
cancer cells in vitro was studied for its anti-apoptotic activity.
METHODS
Apoptotic ratio of the Colo-205 cells was determined after 17-AAG application with terminal deoxynucleotidyl transferase
dUTP nick end labeling (TUNEL) staining and apoptosis related genes. Apoptosis signal path related key mitochondrial
proteins, cytochrome c, bcl-2, caspase 9 and Apaf-1 expression were examined with RT-PCR method.
RESULTS
17-AAG caused induction of cell death. Apoptotic related genes such as cytochrome-c, Apaf-1 and
caspase-9 protein expressions were increased signifi cantly (p < 0.05) and anti-apoptotic bcl-2 expression was
decreased signifi cantly (p < 0.05). Our results indicated that the application of 17-AAG on Colo-205 cells showed
anticancer effect by the apoptosis due to alteration of apoptotic genes.
CONCLUSIONS
The apoptotic effect of 17-AAG as an natural product for alternative medicine would be very
important for the success and quality of life during the treatment of colon carcinoma with the combination of anticancer
drugs.
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S1492
Poster Abstracts – IFCC WorldLab 2017 – Durban, South Africa, 22-25 October 2017 • DOI 10.1515/cclm-2017-7062
Clin Chem Lab Med 2017; 55, Special Suppl, pp S1483 – S1505, October 2017 • Copyright © by Walter de Gruyter • Berlin • Boston
T037
Metabolomics and biomarkers
BLOOD TRANSTHYRETIN LEVEL (TTR): REFLECTION OF CSF TRANSTHYRETIN IN 1ST ONSET DEPRESSIVE
EPISODE: AN EXPLORATORY STUDY.
V. Lal 1
1
All India Institute Of Medical Sciences,Jodhpur,Rajasthan,India
BACKGROUND-AIM
Transthyretin protein is the thyroid hormone binding component that transports thyroxine from blood stream into the
brain. It has been estimated that 3% transthyretin(TTR) in ventricular CSF and 10% TTR in lumbar CSF are derived from
blood. (1) It is expressed in the choroid plexus, and it is the major thyroid hormone –binding protein in the CSF.
Mutations in transthyretin are associated with Bipolar Disorders & depression, familial amyloid polyneuropathy, a
neurodegenerative disorder characterized by TTR deposition in the PNS.Its concentration in plasma is 250
microgram/ml.Each mole of TTR binds 1 mole of T4 with high affinity, the other T4 molecule is bound with low affinity at
higher concentrations of T4. The half-life of TTR in plasma is normally about two days but is less during less during
illness.
The aim of this study was to unravel the role of TTR in CNS physiology that could account for the depressive behavior in
subjects who were euthyroids. The dysfunctional state of TTR by its misfolding and aggregation is known to be
associated with the with neurodegenerative disease like Amyloidosis and Schizophrenia.
METHODS
A sample size of 100, drug naïve cases with varied degrees of 1st onset Depressive episode, and sample size 50 ,age
and gender matched control group were enrolled & all experimental conditions were identically maintained for both case
and control groups.
The objectives of this study are to determine and analyze the
1. Observation on the plausible cause of depression in subjects investigated for routine Thyroid profile
2. Scale Depression by using Hamilton Depression Rating Scale
HDRS Score for the severity of depression:
• 10 – 13 Mild
• 13 – 17 Mild to Moderate
• > 17 Moderate to severe
3.Compare Free Thyroid Hormones with TSH, TBG and Transthyretin in controls and cases and amongst different
grades of severity according to HDRS Scores.
4. Correlating the severity of depression with the level of Blood Transthyretin
RESULTS
The Serum Transthyretin Level(Normal Range 15.7-26.6 mg/dl).There was moderate but significant decrease in
Transthyretin levels in sera of subjects with depression as compared to controls.TTR values in subjects with mild
depression was 13.56mg/dl, with moderate depression was 8.67 mg/dl, and with severe depression was 6.78 mg/dl .
CONCLUSIONS
Though CSF transthyretin is secreted independently in CSF, significant correlation (p=.007)was found between CSF and
Blood transthyretin for same individuals.There was significant correlation within and between the groups as graded by
degrees of depression as measured by HDRS.
Interestingly Jeffrey et al found an ~40% down-regulation of transthyretin levels in post-mortem prefrontal cortex from
patients with schizophrenia compared with controls using Western blot analysis of this peptide.Specificity of this
biomarker signature will need to be assured on a larger set of samples from patients with psychotic depression,Bipolar
disorder and other psychiatric illness.
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Download Date | 10/25/17 2:57 PM
S1493
Poster Abstracts – IFCC WorldLab 2017 – Durban, South Africa, 22-25 October 2017 • DOI 10.1515/cclm-2017-7062
Clin Chem Lab Med 2017; 55, Special Suppl, pp S1483 – S1505, October 2017 • Copyright © by Walter de Gruyter • Berlin • Boston
T038
Metabolomics and biomarkers
PLASMA PHOSPHOLIPIDS AND BRAIN WHITE MATTER HYPERINTENSITIES
D. Li 2, J. Misialek 2, A. Alonso 1
1
Emory University
2
University of Minnesota
BACKGROUND-AIM
Phospholipids such as phosphatidylcholines (PC) and sphingomyelins (SM) are abundant lipid species both in white and
grey matter of the brain. Plasma phospholipids may be used as a biomarker of neurodegeneration.
METHODS
We examined the cross-sectional relationships of 6 plasma metabolites (including 2 PCs and 4 SMs) with brain MRI
measures of neurodegeneration (white matter hyperintensities [WMH]) in 238 participants in the Atherosclerosis Risk in
Communities Neurocognitive Study (ARIC-NCS). WMH were defined as has been codified in recent guideline. All
analysis involving WMH include total intracranial volume as a covariate. Individual metabolites were log-transformed and
standardized. Multivariable linear regression was performed to account for demographics, APOE genotype,
cardiovascular risk factors, and comorbidities.
RESULTS
Lower levels of SM(OH) C24:1 were significantly associated with higher WMH volume (beta [95% confidence intervals] of
0.54 cubic centimeters [-5.49, -0.66] after multivariable adjustments.
CONCLUSIONS
Circulating SM(OH) C24:1 may be a novel, independent plasma biomarker of neurodegeneration.
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Download Date | 10/25/17 2:57 PM
S1494
Poster Abstracts – IFCC WorldLab 2017 – Durban, South Africa, 22-25 October 2017 • DOI 10.1515/cclm-2017-7062
Clin Chem Lab Med 2017; 55, Special Suppl, pp S1483 – S1505, October 2017 • Copyright © by Walter de Gruyter • Berlin • Boston
T039
Metabolomics and biomarkers
QUANTIFICATION OF SERUM HEPCIDIN IN PATIENTS WITH ATHEROSCLEROSIS
V. Manolov 2, S. Hadjidekova 3, J. Petrova 4, V. Vasilev 1, M. Petrova 4, H. Krushkov 4, Y. Jelev 4, P. Jeliazkov 5, Z.
Gramatikova 6, K. Tzatchev 2, L. Traykov 4
1
Clinical laboratory and clinical pharmacology, University “Aleksandrovska” hospital
2
Department of Clinical Laboratory and Clinical Immunology, Medical University Sofia
3
Department of Medical Genetics, Medical University Sofia
4
Department of Neurology, Medical University Sofia
5
Medical University Sofia
6
R.E.D. Laboratories N.V./S.A. – Zellik, Belgium
BACKGROUND-AIM
Hepcidin leads to the deposition of iron in macrophages in atherosclerotic plaques by an increase in lipid peroxidation
and progression of foam cells, which leads to the risk of atherosclerosis. Homocysteine is formed from methionine by
way of the transmetalation. Atherosclerosis and thrombosis are conditions associated with homocystinuria and
significantly high levels of homocysteine in the blood, suggesting that this is related to compromise the integrity of the
vessel wall.
METHODS
45 patients with atherosclerosis were included; 24 females (52.2%). They had clinical and neurological examination,
EMG; IMT and ABI were measured. They were evaluated for routine biochemical parameters, and additional serum
hepcidin and homocysteine were quantified. Hepcidin and homocysteine were evaluated by ELISA methods. The results
obtained from atherosclerosis patients were compared to age and gender matched healthy controls. Statistical analysis
of established results was performed using Pearson’s correlation and Student’s paired t-test.
RESULTS
We found statistically significant elevated serum hepcidin (52.3 µg/L ± 11.5 µg/L) and homocysteine levels (58.8 µmol/L
± 16.7 µmol/L) in atherosclerosis patients compared to healthy controls (19.9 µg/L ± 5.1 µg/L; 8.9 µmol/L ± 1.4 µmol/L);
P<0.005. We found a statistically significant correlation between serum hepcidin and IMT (-0.9<r<-0.7) and ABI
(0.9<r<0.7) in atherosclerosis patients; P<0.005.
CONCLUSIONS
Our findings suggest serum hepcidin quantification as a marker for iron deposition in atherosclerotic plaques and
diagnosis of atherosclerotic changes.
We appreciate financial support of Medical University, Sofia, as this study is part of Project N° 5070/2016 (Contract
4-C/2016) and N° 8082/2016 (Contract Д-54/2017).
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Download Date | 10/25/17 2:57 PM
S1495
Poster Abstracts – IFCC WorldLab 2017 – Durban, South Africa, 22-25 October 2017 • DOI 10.1515/cclm-2017-7062
Clin Chem Lab Med 2017; 55, Special Suppl, pp S1483 – S1505, October 2017 • Copyright © by Walter de Gruyter • Berlin • Boston
T040
Metabolomics and biomarkers
COMPARISON OF TWO METABOLOMIC PROFILING APPROACHES FOR THE ESTIMATING OF COMMUNITY
DRUG ABUSE. UTILITY IN PUBLIC HEALTH AND FORENSIC EPIDEMIOLOGY
B. Moslah 3, E. Hapeshi 2, M. Araoud 3, A. Nouioui 3, S. Najjar 1, D. Amira 3, D. Fatta-Kassinos 2, A. Hedhili 3
1
Faculty of pharmacy, 5000 Monastir, Tunisia
2
International Water Research Center, University of Cyprus, Nicosia, Cyprus
3
Laboratory of Toxicology, Research unit of Environment and Toxicology LR12SP07, Center Mahmoud Yaccoub for
Emergency Medical Assistance, 1008 Tunis, Tunisia
BACKGROUND-AIM
Drug addiction is a topical issue in Tunisia. It is a public health problem with a significant impact on health and society.
METHODS
Two metabolomic profiling approaches were compared by the analysis of illicit drugs in two different matrices, biological
fluids (urine specimens) and in environment samples (wastewater) for the estimation of community drug abuse.
Urine samples were collected from a total of 28298 interpellated individuals suspected to be drug addicts and then
analyzed against a 11-drug panel via fast screening tests EMIT, KIMS. GC-MS was used as analytical tool to obtain a
confirmed result.
Cocaine (COC) and its major metabolite benzoylecgonine (BZE) has been investigated in influent wastewater samples
by UPLC-MS/MS. COC consumption was back-calculated from the measured daily loads of its major metabolite BZE at
the entrance of WWTP.
RESULTS
The results of urine screening tests showed insignificant consumption rate of cocaine (<0.2%). For several years in
Tunisia, we believed that the consumption of cocaine was inconsiderable, although a large illicit drug use happens off-the
police radar. A more realistic picture of local use patterns for the most common illicit drugs was assessed to monitor
changing habits in real time.
Thus, COC consumption was back-calculated from the measured daily loads of its major metabolite BZE at the entrance
of WWTP. An interactive mapping of the real-time cartography consumption of cocaine in the north-eastern (NE) regions
of Tunisia was established. The NE1 region shows the highest consumption with 585 mg/day/1000 inhabitants.
CONCLUSIONS
It is assumed that the wastewater metabolomic approach reflect more closely the actual levels of cocaine consumption
through the analysis of human biomarkers in influent wastewater. The approach could be useful for monitoring in
real-time the community drug abuse, helping scientists and social authorities to combat drug trafficking as well as
protecting human health.
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Download Date | 10/25/17 2:57 PM
S1496
Poster Abstracts – IFCC WorldLab 2017 – Durban, South Africa, 22-25 October 2017 • DOI 10.1515/cclm-2017-7062
Clin Chem Lab Med 2017; 55, Special Suppl, pp S1483 – S1505, October 2017 • Copyright © by Walter de Gruyter • Berlin • Boston
T041
Metabolomics and biomarkers
DIAGNOSTIC PERFORMANCE OF ENHANCED LIVER FIBROSIS (ELF) TEST IN PREDICTING LIVER STIFFNESS
E. Nah 1, S. Cho 1, S. Kim 1, H. Cho 2
1
Department of Laboratory Medicine and Health Promotion Research Institute, Korea Association of Health Promotion,
Seoul
2
MEDIcheck LAB, Korea Association of Health Promotion, Cheongju
BACKGROUND-AIM
Fatty liver disease is not uncommon in general population. It represents a wide spectrum of pathologic findings, from
simple steatosis to steatohepatitis, fibrosis, cirrhosis, and hepatocellular carcinoma. The diagnostic assessment of liver
fibrosis is an important step not only in the management of patients with chronic liver diseases but also in the
assessment of the true burden of liver disease in the general population. Liver biopsy is considered the gold standard for
the assessment and quantification of liver fibrosis. But noninvasive techniques including serum biomarkers have been
developed to circumvent the need for liver biopsy. Therefore, the aim of our study is to assess the diagnostic
performance of enhanced liver fibrosis (ELF) test in predicting liver stiffness by using magnetic resonance elastography
(MRE) as a reference standard in health checkups.
METHODS
This study included 89 health examinees who underwent MRE and ELF test at health promotion center in Korea,
between July 2016 and December 2016. ELF score was compared with MRE results. Receiver operating characteristic
(ROC) curve analysis was performed for ELF test as a predicting test for liver stiffness.
RESULTS
Area under ROC (AUROC) to predict mild liver stiffness, and moderate-to-severe liver stiffness were 0.613, and 0.891 for
ELF test. Optimized cutoffs of ELF to maximize sum of sensitivity and specificity were 8.98, and 10.3 for mild stiffness,
and moderate-to-severe stiffness, respectively.
CONCLUSIONS
ELF test demonstrated considerable diagnostic value in predicting liver stiffness in health checkups.
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S1497
Poster Abstracts – IFCC WorldLab 2017 – Durban, South Africa, 22-25 October 2017 • DOI 10.1515/cclm-2017-7062
Clin Chem Lab Med 2017; 55, Special Suppl, pp S1483 – S1505, October 2017 • Copyright © by Walter de Gruyter • Berlin • Boston
S1498
T042
Metabolomics and biomarkers
STUDY ON EFFECT OF WET CUPPING (HEJAMAH) ON BLOOD LIPID PROFILE IN HUMAN
ALDYSSAH-ALSHATI LIBYA
AT
G. Alajwad 1, A. Elwafa 1, A.F. Hawad 3, A.M. Lalem 2, A. Nouh 2
1
Faculty of Engineering and Technology, Sebha University, Libya
2
Faculty of Medical Technology, Marzouk, Sebha University, Libya
3
Faculty of Medicine, Sebha University, Libya
BACKGROUND-AIM
This study was aimed to evaluate the impact of Hejamah on blood lipid profile in hyperlipidaemias human at
Aldyssa-Alshati Town, Libya.
METHODS
It was conducted on 32 volunteers, from which 9 volunteers were males and 23 were females, their ages range was from
22 to 77 years. Two samples were taken from each volunteer, the first one was taken before cupping and the second
sample was taken one week later after cupping. Serum was used for determination of the levels of lipid profile, (Total
cholesterol (T. Cho), triglyceride (TG), high-density lipoproteins (HDL) and low-density lipoproteins (LDL)).
RESULTS
The results have shown a significant decrease in both the levels of serum T. Cho and TG, but no changing in the levels
of HDL and LDL, in hyperlipidaemias male and female volunteers after cupping.
Table (1):Show the concentration of serum lipid profile (Total Cho., TG, HDL & LDL) in male volunteers before and after
cupping.
Parameters n=9 Before cupping (mean ±SD) One week after cupping (mean ±SD) P- Value
T.Cho (mg/dL) 177.3±40.2 167.0±37.4 0.238
T G (mg/dL) 150.6±63.7 122.6±58.1 0.124
HDL (mg/dL) 38.1±7.8 37.2±6.0 0.615
LDL (mg/dL) 130.9±36.1 131.3±39.7 0.938
Table(2): Show the concentration of serum lipid profile (Total Cho., TG, HDL & LDL) in female volunteers before and
after cupping.
Parameters (n =23) Before cupping (mean ±SD) One week after cupping (mean ±SD) P- Value
T.Cho (mg/dL) 188.5±49.4 177.3±34.9 0.159
T G (mg/dL) 141.2±65.8 131.8±77.9 0.571
HDL (mg/dL) 47.9±10 45.7±13.1 0.402
LDL (mg/dL) 147.8±57.8 139.5±38.6 0.315
Table 3: Show the eves of blood lipid profile (Total Cho., T G, HDL & LDL) in hyperlipidemic volunteers.
Parameters (n= 11) Before cupping (M ±SD) A week after cupping (M ±SD) P Value
T.Cho (mg/dL) 235.3±34.5 202.5±24.0 0.012*
T G (mg/dL) 171.0±58.0 136.7±54.3 0.041*
HDL (mg/dL) 49.9±10.27 43.0±11.6 0.108
LDL (mg/dL) 190.7±71.0 167.2±39.6 0.089
CONCLUSIONS
The Hejamah approach, have an impact on decreasing the levels of serum, T. Cho and TG, and for that Hejamah
(cupping) might be cardioprotective, and also wet cupping could represent a prospective complementary therapy for the
hyperlipidaemic patient.
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Download Date | 10/25/17 2:57 PM
Poster Abstracts – IFCC WorldLab 2017 – Durban, South Africa, 22-25 October 2017 • DOI 10.1515/cclm-2017-7062
Clin Chem Lab Med 2017; 55, Special Suppl, pp S1483 – S1505, October 2017 • Copyright © by Walter de Gruyter • Berlin • Boston
T043
Metabolomics and biomarkers
TUMOUR NECROSIS FACTOR ALPHA AND OXIDATIVE DNA DAMAGE IN CHRONIC EXPOSURE TO CEMENT
DUST
A.C. Nsonwu-Anyanwu 2, L.T. Obaji-Ogar 1, E.R. Egbe 1, S.J. Offor 1, C.A.O. Usoro 1
1
Unit of Chemical Pathlogy, Department of Medical Laboratory Science, University of Calabar, Cross River State, Nigeria.
2
Unit of Chemical Pathology, Department of Medical Laboratory Science, University of Calabar, Cross River State,Nigeria
BACKGROUND-AIM
Inflammatory cell activation, oxidative stress and oxidative DNA damage have been associated with exposure to cement
dust; heavy metals present in cement dust have been implicated. This study evaluates the biomarkers of oxidative stress
and inflammation and heavy metals in relation to duration of exposure to cement dust among cement workers.
METHODS
Ninety consenting apparently healthy male subjects aged 18-60 years comprising of 45 cement workers and 45
non-cement workers were studied. The tumor necrosis factor alpha (TNF-) and urinary 8-hydroxy-2-deoxyguanosine
(8-OHdG) were estimated by ELISA methods, biomarkers of oxidative stress (malondialdehyde (MDA), glutathione
(GSH), nitric oxide (NO), total antioxidant capacity (TAC), total plasma peroxides (TPP)), and uric acid (UA) were
determined using colorimetric methods, heavy metals (Arsenic (As), Chromium (Cr), Cadmium (Cd)) were determined by
atomic absorption spectrophotometry while oxidative stress index (OSI) was obtained by calculation. Anthropometric
indices, blood pressure and socio-demographic information were obtained using standard methods. Data were analysed
using t-test, ANOVA, LSD post hoc and Pearson’s correlation at p <0.05.
RESULTS
The MDA, TPP, OSI, 8-OHdG, TNF-, As and Cr levels were significantly higher and UA, TAC and GSH lower in cement
workers compared to non-cement workers. Higher levels of Cd, MDA, 8-OHdG and TNF- and lower levels of GSH were
observed with increasing duration of exposure to cement dust. Positive correlation was observed between 8-OHdG and
TNF- (r = 0.492, p = 0.001) and negative correlation between GSH and MDA (r = -0.490, p = 0.001) only in cement
workers.
CONCLUSIONS
Chronic exposure to cement dust is associated with depletion of antioxidants, increased lipid peroxidation, oxidative
stress and oxidative DNA damage which may be implicated in the development of chronic lung conditions.
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S1499
Poster Abstracts – IFCC WorldLab 2017 – Durban, South Africa, 22-25 October 2017 • DOI 10.1515/cclm-2017-7062
Clin Chem Lab Med 2017; 55, Special Suppl, pp S1483 – S1505, October 2017 • Copyright © by Walter de Gruyter • Berlin • Boston
T044
Metabolomics and biomarkers
PREVALANCE OF VITAMIN D DEFICIENCY IN ADULTS IN KONYA, TURKEY
E. Paydas Hataysal 1
1
SELCUK UNIVERSITY, FACULTY OF MEDICINE ,Department of Biochemistry
BACKGROUND-AIM
Vitamin D is well known as a hormone involved in mineral metabolism and growth of bone. Deficiency of vitamin D
results in impaired formation of bone, producing rickets in children and osteomalacia in adults. Vitamin D is important for
many metabolic pathways and different health outcomes and the physiology of vitamin D is very complex. Vitamin D3 of
the skin is hydroxylized in the liver into 25-hydroxyvitamin D [25-(OH)D] and subsequently in the kidney by the
1-hydroxylase into its active metabolite 1,25-dihydroxyvitamin D. Multiple factors like ethnicity, age, sex, diseases, and
medication influence vitamin D concentrations. Vitamin D deficiency is defined as a serum concentration of ≤20 ng/mL.
This study aimed to determine prevalance of vitamin D deficiency among adults people in Konya region of Turkey.
METHODS
The 25(OH) vitD3 test results of 1037 patients who applied to Selcuk University Medical School Hospital between the
years of 2014 and 2016 were collected retrospectively. Patients with chronic disease and inflammatory disorders were
excluded. Vitamin D levels were measured by a choromotographic method(LC/MS/MS) with API3200. Statistical analysis
was performed with IBM SPSS v20.
RESULTS
Vitamin D levels of 134 ( %12.9) patients were <5 ng/mL. Vitamin D levels of 479 (%46.1) patients were <10 ng/mL .
Vitamin D levels of 854 (%82.3) patients were <20 ng/mL
CONCLUSIONS
The results indicate that vitamin D deficiency is common in Turkish society. Vitamin D reference values should be
redetermined for the Turkish population.
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S1500
Poster Abstracts – IFCC WorldLab 2017 – Durban, South Africa, 22-25 October 2017 • DOI 10.1515/cclm-2017-7062
Clin Chem Lab Med 2017; 55, Special Suppl, pp S1483 – S1505, October 2017 • Copyright © by Walter de Gruyter • Berlin • Boston
T045
Metabolomics and biomarkers
RE-ASSESSMENT AND OPTIMIZATION OF AN ORGANIC ACID METHOD FOR AUTOMATION
M.M. Phiri 1
1
North-West University, Department of Biochemistry, Potchefstroom Campus.
BACKGROUND-AIM
Sample preparation is a necessary prerequisite for GC/MS analysis of urinary organic acids for clinical diagnosis of
inborn errors of metabolism (IEM). The sample clean-up step in the analytical process poses a challenge. It involves the
isolation of the analytes of interest by an extraction process from a complex matrix to one that is more suitable for the
analytical platform. As opposed to a fully automated and high-throughput sample preparation protocol, the existing
urinary organic acid extraction method in many laboratories worldwide is still performed manually. It is labour-intensive
and time-consuming, requires multiple exhaustive pipetting steps, uses large amounts of toxic solvents that can be
hazardous to health and needs trained and expert personnel to operate the method. The literature documents the
progress made in miniaturizing and automating the solvent extraction, but scarce literature is available on how this has
been applied to the urinary extraction of organic acids. Thus, the development of a method that can be fully automated
would improve the sample throughput and eliminate the intense labor involved, simplify the method so that even
non-specialists can perform it, thus eliminating most of the other setbacks associated with manual extraction. Therefore
the aim of the study was to reassess the in-house organic acid extraction method and optimize it for automation.
METHODS
The experimental workflow involved the selection of an initial suitable miscible solvent for rapid extraction of organic
acids. The other requirement for this miscible solvent was that it would enable better extraction of very polar organic
acids. This was followed by the selection of a suitable immiscible solvent that would ensure good isolation of organic
acids, quick evaporation and clear phase separation that would render centrifugation unnecessary. The solvent ratios
and volumes were optimized and miniaturized to small volumes of solvents. The miniaturized organic acid protocol was
translated into a fully automated extraction procedure on a liquid AutoSampler. The automated method was validated for
linearity, precision, recovery and accuracy.
RESULTS
A two-phase extraction system using two optimal solvents, acetonitrile, and ethyl acetate, were found to be efficient in
the extraction of urinary organic acids. It enabled efficient and rapid extraction. The analytical range of the method for
most of the analytes was established to be between 1 – 500 mg/l. The correlation coefficient (r) of all analytes was
generally > 0.99, with a few exceptions. The analytical ranges of the specific analytes showed that the test results within
these ranges are reliable and can be reported. The repeatability was generally below 20% but had higher
within-laboratory precision. The automated method’s overall imprecision was better than the in-house method. The
accuracy of the method was determined by a method comparison experiment with ERNDIM EQAS samples for
quantitative organic acids. The mean of the test results was compared to the mean of all the laboratories. The
proportional systematic error of the method ranged from -0.18 to 2.06. The constant systematic error for the analytes was
-5.75 and 5.66. The total error of the method determined demonstrated a reduction in random and systematic errors
when compared to the current in-house manual method. It was also noted that the correlation coefficient between the
new method and the expected results was substantially better when compared to the current in-house method. By
implication, the regression model fit was substantially better for the automated method. This created an opportunity for
bias correction through the use of extraction factors, instrument response factors, the use of external calibration curves
or reassignment of standard/calibrator concentrations for the new method as opposed to the current in-house method
where this was not an option.
CONCLUSIONS
Based on the findings in this study, it was concluded that an automated procedure for LLE of urinary organic acids was
successfully developed. The goal of having a method that could give consistent extraction and meet the criteria for
automation was achieved. All the extraction steps were optimized and the method proved to have good extraction
efficiencies for organic acids and to improve the performance of the existing in-house method. The automated method
can significantly reduce the total cost of the analysis of organic acids. It also would enable non-specialised laboratories to
analyze organic acids as well.
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Download Date | 10/25/17 2:57 PM
S1501
Poster Abstracts – IFCC WorldLab 2017 – Durban, South Africa, 22-25 October 2017 • DOI 10.1515/cclm-2017-7062
Clin Chem Lab Med 2017; 55, Special Suppl, pp S1483 – S1505, October 2017 • Copyright © by Walter de Gruyter • Berlin • Boston
T046
Metabolomics and biomarkers
INFLUENCE OF TWO INTRAVENOUS IRON FORMULATIONS ON ISOPROSTANE LEVELS IN HEMODIALYZED
PATIENTS
J. Racek 2, J. Eiselt 1, D. Rajdl 2, V. Senft 2, L. Trefil 2
1
1st Dept. of Medicine, Faculty Hospital and Faculty of Medicine, Charles University, Pilsen, Czech Republic
2
Dept. of Clinical Biochemistry and Hematology, Faculty Hospital and Faculty of Medicine, Charles University, Pilsen,
Czech Republic
BACKGROUND-AIM
Patients on chronic hemodialysis (HD) treatment need intravenous administration of iron. However, iron as a transient
metal can participate in hydroxyl radical production in Fenton´s reaction. The aim of our intervention study was to
compare two intravenous iron formulations administered to HD patients with respect to oxidative stress induction.
METHODS
We enrolled 10 HD patients (median age 70.3 years [range 21 to 79], 3 women, median 26 months on HD [range 7 to
263 months]) that sequentially received one of the intravenous iron formulations (iron sucrose – Venofer, ferric
carboxymaltose – Ferinject) or placebo in a crossover design. The iron dose was 200 mg administered in 100 ml of
physiological saline in a 30-minute infusion that began after 60 minutes of HD procedure. We have determined serum
iron (routine spectrophotometric method and atomic absorption spectrometry, AAS), serum isoprostanes (ELISA,
Cayman Chemical) and TBARS (spectrophotometrically) before HD and after 60, 90, 150 (i.e. after iron infusion), 240
minutes after beginning of HD procedure, and finally before the next HD. Differences of serial measurements among
individual iron formulations were compared using area under the curve (AUC) comparisons by ANOVA.
Student-Newman-Keuls test was used for post-hoc pairwise comparisons.
RESULTS
Isoprostane levels AUCs were significantly higher after administration of Venofer (and not Ferinject) in comparison with
placebo (p<0.05). Serum iron concentration AUCs measured by AAS were clinically comparable between the
formulations (Ferinject 1.2 times higher values than Venofer, p<0.05), whereas routine spectrophotometric method gave
clearly lower (1.9 times) concentrations after Ferinject vs Venofer administration (p<0.05). The concentrations of TBARS
significantly correlated with plasma iron concentration measured by AAS (r=0.81, p<0.001).
CONCLUSIONS
Venofer (and not Ferinject) seems to promote oxidative stress after i.v. application. Administration of the same iron dose
(200 mg) leads to clearly different iron concentrations after i.v. Venofer and Ferinject administration measured by a
routine spectrophotometric; that is why this method cannot be used for evaluation of iron status after i.v. iron
administration. Measurement of TBARS is strongly influenced by iron concentration in the sample and measured values
are probably artefact after administration of i.v. iron.
Supported by MH CZ-DRO (Faculty Hospital in Pilsen – FNPl, 00669806)
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S1502
Poster Abstracts – IFCC WorldLab 2017 – Durban, South Africa, 22-25 October 2017 • DOI 10.1515/cclm-2017-7062
Clin Chem Lab Med 2017; 55, Special Suppl, pp S1483 – S1505, October 2017 • Copyright © by Walter de Gruyter • Berlin • Boston
T047
Metabolomics and biomarkers
ANALYTICAL AND CLINICAL VALIDATION OF CHEMILUMINISCENT ASSAY FOR HEPCIDIN-25 DETERMINATION
M. Savković 1, D. Ćujić 3, S. Simić-Ogrizović 2, V. Dopsaj 1
1
Center for Medical Biochemistry, Clinical Center of Serbia, Belgrade, Serbia; Faculty of Pharmacy, University of
Belgrade, Belgrade, Serbia
2
Clinic of Nephrology, Clinical Center of Serbia, Belgrade, Serbia; School of Medicine, University of Belgrade, Serbia
3
Institute for The Application of Nuclear Energy INEP, University of Belgrade, Belgrade, Serbia
BACKGROUND-AIM
Iron metabolism is under control of hepcidin-25 (Hep), a peptide hormone synthesized in the liver. As biomarker, Hep
could be useful in diagnosis and follow-up of iron-related anemias. Although Hep could be measured in serum and urine,
Hep determination is not yet widely used in clinical routine. Mass spectrometry based methods are reliable, but
expensive, require well-trained staff and therefore are available to limited numbers of laboratories. Immunochemical
assays are more affordable and easier to perform, but there are problems regarding standardisation and specifity.
The aim of this study was to assess analytical and clinical performances of commercially available imunochemical assay
for Hep determination.
METHODS
The Hep concentration in human serum is determined with HEPCIDIN-25 Chemiluminiscent Direct ELISA assay
(Corgenix, CO, USA).This test is based on pair of monoclonal antibodies specific to Hep, and the intensity of
chemiluminiscent signal is proportional to the concentration of Hep in sample. The Hep concentration was determined in
sera of healthy persons (control, n=28), and in sera of patients with iron metabolism disorders: iron defficiency anemia
(IDA group, n= 34) and hemodialysis-dependent patients with chronic kidney disease (HD group, n=111).
RESULTS
The concentration range covered with standard curve was 0-406 ng/mL. The reproducibility of measurement (expressed
as coefficient of variation) was 8.0 % for intra-assay and 17.8 % for inter-assay measurements. The linearity of
measurement was assessed with serial dilutions of human serum sample with high Hep concentration. The mean Hep
concentration in control was 15.3 ng/mL, while levels were lower (p < 0.05) in women than in men, 8.7 and 29.3 ng/mL
respectively. In all IDA subjects, low Hep levels were measured, 6.9 ng/mL. In contrast to this, elevated Hep levels were
found in CKD patients, 74.5 ng/mL. The differences in Hep levels between male and female patients with iron
metabolism disorders were not observed.
CONCLUSIONS
The results of analytical and clinical validation suggest that concentration of Hep in human serum could be reliably
measured using chemiluminiscent immunoassay, which therefore might be used as diagnostic tool in patients with
anemia due to iron disorders.
Unauthenticated
Download Date | 10/25/17 2:57 PM
S1503
Poster Abstracts – IFCC WorldLab 2017 – Durban, South Africa, 22-25 October 2017 • DOI 10.1515/cclm-2017-7062
Clin Chem Lab Med 2017; 55, Special Suppl, pp S1483 – S1505, October 2017 • Copyright © by Walter de Gruyter • Berlin • Boston
T048
Metabolomics and biomarkers
SERUM BILIRUBIN LEVELS AND PROMOTER VARIATIONS IN HMOX1 AND UGT1A1 GENES IN PATIENTS WITH
FABRY DISEASE
A. Jiraskova 1, G. Bortolussi 1, G. Dostalova 1, L. Eremiasova 1, L. Golan 1, V. Danzig 1, A. Linhart 1, L. Vitek 1
1
Charles University, Prague
BACKGROUND-AIM
The aim of our study was to assess the possible relationships among heme oxygenase (HMOX), bilirubin
UDP-glucuronosyl transferase (UGT1A1) promoter gene variations, serum bilirubin levels, and Fabry disease (FD) (a
rare lysosomal storage disease characterized by impaired metabolism of neutral glycosphingolipids).
METHODS
The study included 56 patients with FD (M:F ratio = 0.65) and 185 healthy individuals. Complete standard laboratory and
clinical work-up was performed on all subjects, together with the determination of total peroxyl radical scavenging
capacity. The (GT)n and (TA)n dinucleotide variations in the HMOX1 and UGT1A1 gene promoters, respectively, were
determined by DNA fragment analysis.
RESULTS
Compared to controls, patients with FD had substantially lower serum bilirubin levels (12.0 vs. 8.85 umol/L, p=0.003),
and also total peroxyl radical scavenging capacity (p<0.05), which showed a close positive relationship with serum
bilirubin levels (p=0.067) as well as with the use of enzyme replacement therapy (p=0.036). The presence of the L allele
in the HMOX1 gene promoter, responsible for lower enzyme activity, was not associated with the manifestation of FD
(OR=1.69; 95%CI=0.54-5.24, p=0.46). In contrast, the presence of the TA7 allele UGT1A1 gene promoter, responsible
for higher systemic bilirubin levels, was associated with a two-fold lower risk of the manifestation of FD (OR=0.51,
95%CI=0.27-0.97, p=0.038).
CONCLUSIONS
FD is associated with markedly lower serum bilirubin levels, most likely due to bilirubin consumption during increased
oxidative stress accompanying FD. However, genetic factors affecting defense against increased oxidative stress
(UGT1A1 promoter gene polymorphism) seem to modify the manifestation of FD as well.
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Download Date | 10/25/17 2:57 PM
S1504
Poster Abstracts – IFCC WorldLab 2017 – Durban, South Africa, 22-25 October 2017 • DOI 10.1515/cclm-2017-7062
Clin Chem Lab Med 2017; 55, Special Suppl, pp S1483 – S1505, October 2017 • Copyright © by Walter de Gruyter • Berlin • Boston
T049
Metabolomics and biomarkers
ROLE OF PARVOVIRUS B19 IN ANEMIC SYNDROME DURING ABNORMAL PREGNANCY
S. Voleva 3, V. Manolov 2, S. Ivanova 3, B. Marinov 5, S. Angelova 3, V. Vasilev 1, S. Shishkov 4
1
Clinical laboratory and clinical pharmacology, University “Aleksandrovska” hospital
2
Department of Clinical Laboratory and Clinical Immunology, Medical University
3
Department of Virology, National Centre of Infectious and Parasitic Diseases (NCIPD)
4
Laboratory of Virology, Faculty of Biology, St. Kl. Ohridski Sofia University
5
University Obstetrics and Gynecology Hospital “Maichin Dom”
BACKGROUND-AIM
Viral infections during pregnancy are a leading cause of severe complications and mortality of mother and fetus. This
study aims to analyze the participation of parvovirus B19 as a etiologic agent in the development of anemic syndrome,
support a proper differential diagnosis and improve prognostic and treatment of women with abnormal pregnancies and
newborns in Bulgaria.
METHODS
We examined 57 serum samples of pregnant women with abnormal pregnancy, treated in SBALAG “Maichin Dom” Sofia. Patients were divided into four groups: pregnant women with anemia (n=21; 36.8%), non-immune hydrops fetalis
(n=7; 12.3%), fetal ascites (n=6; 7.0%) and women suffered a miscarriage (n=25; 43.9%). Serological (indirect ELISA
test) and molecular (B19V-PCR test) methods were used. CLIA, ELISA and NEPH methods were included to determined
indicators of iron homeostasis among patients with anemia.
RESULTS
A total of 21/57 (36.8%) patients were confirmed with primary reactive B19V-IgM antibodies - the rate of detected
B19V-IgM antibodies was as follows: pregnant women with anemia (33.3%), non-immune hydrops fetalis (19.0%), fetal
ascites (4.8%) and miscarriage (42.9%). Protective B19-IgG antibodies were found in 35/57 (61.4%) samples. Anemia
was determined as iron-deficient according to the low serum levels of hepcidin 2.3 ± 0.3 g/L, compared to pregnant
women with no anemia 18.4 ± 2.8 g/L; P<0.001.
CONCLUSIONS
Molecular-diagnostic approach for analyzing B19V showed the highest rate of engagement of that virus in the
development of abnormal pregnancy in women with anemic syndrome. Determination of serum hepcidin in pregnant
women with parvovirus B19 infection would help to clarified the etiological impact of the anemic syndrome and prevents
improper supplementation with iron during pregnancy.
We kindly appreciate financial support from Medical University – Sofia, as this project is part of Grant 2017 – Contract
Д-124/2017.
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Download Date | 10/25/17 2:57 PM
S1505
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