Clinica Chimica Acta -...

Gastrointestinal diseases, including hepatic and pancreatic diseases

T374

Morphological liver structure (light and microscopic observation)and immunoglobulin concentration in serum during disturbedlymph flow from the liver in rats

B. BeckScientific and Diagnostic Medical Center BB-Med, Ruda Slaska, Poland

Background-aim

Liver fibrosis plays a pivotal role in liver function impairment.Hepatic fibrosis is a complex process that involves changes in theamounts of extracellular matrix components, activation of cellscapable of producing matrix materials, cytokin release, and tissueremodeling. The majority of patients have an increase in IgGconcentration, and in cases of inflammatory-necrotic exacerbationsalso IgM. Hepatic lymph flow - example of regulating the lymphaticmicroenvironment on the cellular and functional relationship withthe nervous and endocrine system, is still interesting and unex-plained the object of studies. Little is know about the alteration ofmetabolic pathways of the liver tissue during disturbed lymph flow.

Methods

Male albino Wistar rats (162 animals) weighing between 200 and230 g were selected for the experiment. The animals were kept instable condition and were fed a standard diet with no fluidrestriction. The animals were divided into 9 experimental groups.In each experimental group were 3 subgroups: Group K was thecontrol group – sham operated rats. Group B consisted of rats withdisturbed lymph flow from a liver. Group 0 – animals not subjectedto any surgery, but were placed under pentobarbital anaesthesia.Each subgroup consisted of 6 rats. Lymphostasis was induced ingroup B rats by tying two silk ligatures passed behind the hepatictrunk just distal to the juncture of the hepatic ducts.

The rats were sacrificed for experiment in 1, 3, 7, 14, 21, 28, 35,56 and 103 day after operation to obtain blood samples directly fromthe heart. Serum concentration of immunoglobulins was measured(radial immunodiffusion, Binding Site). Differences between groupswere assessed by ANOVA and RIR Tukey test, and were consideredsignificant when pδ0.05.

Liver tissue was studied for light and electron microscopy.

Results

Our results showed appearance of collagenous fibres in heat-ocytes. We observed an increase of IgG concentration in serum. Theconcentration of IgM not changed.

Conclusions

Disturbed lymph flow from a liver contributes to liverfibrogenesis and probably to hepatocirrhosis. During disturbedlymph flow from the liver immune mechanisms associated with Blymphocytes are hindered and there is disorders in the primaryimmune response.

doi:10.1016/j.cca.2019.03.762

^T375

Laboratory assessment of total bilirubin assay in hospitalizedadult patients and incidence of Gilbert's syndrome in Saudi Arabia

W. Tamimi, N. Alotabi, A. Alsughair, T. Bautista, A. AlhamadKing Abdulaziz Medical city, KIng Saud Bin Abdulaziz University forHealth Sciences, King Abdullah International Research Center, Riyadh,Saudi Arabia

Background-aim

Bilirubin is produced from the breakdown of hemoglobin inbone marrow, the spleen, and other organs. It is carried out to theliver in the blood where it conjugated and excreted into the bile.High levels of bilirubin in the blood may produce jaundice andindication for other liver disease. Gilbert's syndrome is aninherited disorder. It is estimated that between three to 7% of alladults have Gilbert's syndrome. It is much more common in males,and usually first appears when people are in their teens or earlyadulthood. In this study we tried to assess the total bilirubin assayin calculating the incidence of Gilbert's syndrome in Saudipopulation.

Methods

A total of 2276 adults patients were visiting our primary healthclinics in Riyadh, Saudi Arabia during January 2018. These patientshave been selected as they had no serious liver diseases. The bloodwas drawn and collected in plain tube from Becton Dickenson.Samples were transported to the lab and centrifuged for 10 min at3000g

^. The samples were transported to the analyzers to measure

bilirubin, ALT and AST. Bilirubin was measured by diazo reagent toform the colored compound azobilirubin using the chemistryanalyzer Architect 16,000 from Abbott diagnostic. ALT and AST wereperformed on an enzymatic methods on the same analyzer. Thestatistical date were analyzed using Microsoft excel sheet and thediagnostic sensitivity and specificity were calculated.

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Clinica Chimica Acta 493 (2019) S355–S378

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Results

The incidence of observed Gilbert's syndrome in Saudi populationwas found to be 4.1% The sensitivity and specificity values forbilirubin compared to other liver function tests were found to be 28%and 95% respectively. The positive (PPV) and negative (NPV)predictive values were found to be 47% and 90% respectively. Theoverall diagnostic accuracy for total bilirubin was found to be 86.8%.

Conclusions

The colorimetric total bilirubin assay has lower sensitivity butbetter specificity compared to other enzymatic assay of ALT and ASTin evaluating patients for Gilbert's syndrome in Saudi population.Further studies with other sensitive bilirubin methods such as HPLCmay be needed.

doi:10.1016/j.cca.2019.03.763

T376

Evaluation of the implementation of a stool antigen assay fordetecting helicobacter pylori infection in our health area

E.J. Laserna-Mendietaa, C. Andrés-Fernándeza, M. Esteso-Peronaa, M.Montealegre-BarrejónbaClinical Laboratory, Hospital General de Villarrobledo, Villarrobledo,SpainbDepartment of Gastroenterology, Hospital General de Villarrobledo,Villarrobledo, Spain

Background-aim

Helicobacter pylori (HP) is the most prevalent chronic bacterialinfection and is associated with peptic ulcer disease, chronic gastritis,gastric adenocarcinoma, and gastric mucosa associated lymphoidtissue lymphoma. One of the non-invasive tests employed in clinicallaboratories to detect HP infection is stool antigen assay. Thepresence of HP antigen in stool indicates an ongoing infection andthus it can be used both to establish the initial diagnosis and toconfirm its eradication after treatment.

Our aim is to perform a descriptive study of the stool HP antigenassay results in our health area (composed by 65,000 patients) sinceits implementation at the beginning of 2017.

Methods

Demographical characteristics from all the stool HP antigen assayrequests to our laboratory in 2017 and 2018 were obtained. HPantigen in stool samples was determined using a rapid lateral flowimmunochromatographic assay. Contingency tables were generatedfor the following variables: year of petition, gender, age andpetitioner service. GraphPad v5.0 was used to perform statisticalanalysis using chi-square test.

Results

The overall results were 3037 petitions, 1494 in 2017 and 1543 in2018. Positive results were 22.1% and negative results were 70.8%,while 7.1% of them were not processed (mostly due to not providinga stool sample). A significant higher percentage of positive resultswere observed in 2018 (26.2% vs 17.9%, p b .001). Female genderrepresented 65.0% of all petitions, with a similar percentage of

positive results to those in males (23.1% vs 20.3%). Likewise, nodifferences in results were found among four ranges of ages (δ18,19–50, 51–65 and ε66) and the higher percentage of requests wereobserved for the 19–50 year old group (46.7%). The petitionerservices that demanded more frequently the test were generalpractitioners (25.3%), gastroenterology consultants (17.3%) andalergology specialists (8.4%). We found statistical differences amongthem (p b .001), as the petitions from gastroenterologists showed thehighest ratio of negative results (80.8%) and the lowest percentage ofnot processed samples (2.5%). This could be explained by the factthat gastroenterologists are in charge of establishing HP eradicationtreatment.

Conclusions

The stool HP antigen assay has become a highly demanded test inour area, with a mean of 1518 determinations per year. This test ismore requested for adult women (between 19 and 50 years old)attended by general practitioners and gastroenterology consultants.The percentage of positive results was not influenced by gender orage, and it was significantly higher in the second year ofimplementation.

doi:10.1016/j.cca.2019.03.764

T377

Laboratory prevalence data of helicobater pilory infection

P. Argente Del Castillo Rodríguez, D. Morell-García, M.A. BallesterosVizoso, L. Valiña Amado, J.A. Delgado Rodríguez, A. Rubio Alaejos, M.B. Badal Cogul, J.M. Bauça, A. García RajaDepartment of Laboratory Medicine, Hospital Universitari Son Espases,Palma de Mallorca, Spain

Background-aim

Helicobacter pylori (Hp) plays a key role in the development ofdigestive diseases. It affects 50% of the general population (25–40% inEurope). The incidence has been decreasing in association withhealth improvements and eradication's methods. The aim is toevaluate the percentage of positive results in 13C-labeled urea breathtest for Hp detection (UBT).

Methods

A retrospective observational study was performed in a tertiaryhospital between January 2015 and December 2018, analyzing thetotal requests for UBT. It measures by infrared spectrophotometry inPOCone (Otsuka, Japan) considering a positive result when 13C (100mg) enrichment in exhaled air is ε 2,5%, expressed an absolutedifference of 13CO2 [%].

We stratify the requests according to sex and age groups (b30,30–50, N50-year-old). Results were compared using the Chi-squaretest and bivariate analysis, type ⟨ error: 5%.

Results

Total made requests have increased in the last four years by 16%[range:(−)14–39%], percentage of positive results has decreased by2% [range:(−)9–2%] (p= .047).

Total positive percentages were stratified by sex and notsignificant differences were found (34.6% male vs 33.6% female).

Abstracts / Clinica Chimica Acta 493 (2019) S355–S378

b30 years group accounted for 31.95% of positives, 30–50 yearsgroup 35.55% and N 50 years group 33.02% (p = .002).

Total request was adjusted by sex and age, finding significantdifferences in women and highest percentage was observed at 30–50 years group (36.01%) [p = .001].

Women groups were analyzed by Odds ratio and 0.79 [95%CI:0.68–0.96](p= .018) was found at b30 years group.

Conclusions

During the last four years has been an increase in the totallymade requests to the gastroenterology laboratory, highlighting thedecrease in the number of positives.

Moreover, like previous authors has described, being a womanyounger than 30 years old seems to be a protective factor for apositive test. It is probably due to protective effects of estradiol andprogesterone on gastric mucosal response to Hp infection. However,highest incidence was observed in 30–50 years women group.

Slow decrease in the incidence and prevalence of Hp could be dueto the increase in antibiotic resistance, representing an obstacle to itseradication. Follow-up the prevalence's evolution of Hp is necessaryfrom the Laboratory to improve the infection control strategies.

doi:10.1016/j.cca.2019.03.765

T378

Review of transaminase biological reference intervals at ourmedical laboratory

D. González Benito, S. García Castañón, C. Sopeña Sánchez, F.J. CepedaPiorno, C. Alberdi García Del Castillo, V. García Moreira, A. LlorenteTorres, E. Fernández RodríguezClinical Analysis Department, University Hospital of Cabueñes, Gijón,Spain

Background-aim

Liver transaminases ALT and AST are useful biomarkers of somedegree of injury in liver function. At our laboratory, these parametersare measured with an automated analyzer ADVIA 2400 ChemistrySystem (Siemens Healthineers). The medical technology companyrecommendations on reference intervals (RIs) are: 10-40 U/L formales and 7-35 U/L for females for ALT; and for AST are 8-20 U/L inboth genders.

The aim of this study is to define the RIs for these enzymes inadult population at our health area.

Methods

The samples were selected from apparently healthy peopleduring the last two years from January 2016 to December 2017.The average age of this population was 48 years with an age rangebetween 18 and 91 years.

Statistical analysis was performed with MedCalc v11.2. Variabledistribution was studied through graphs and normality was testedwith Kolmogorov-Smirnov. Consequently, the RIs and 95% confi-dence intervals were calculated using nonparametric method(2.5th–97.5th percentile). We used t-Student test to examinedifferences between men and women. A p-valueb.05 was consid-ered significant.

Results

The results were as follows:ALT value for men (U/L): n= 846; Minimum:9; Maximum:66;

Median(CI90%):23(23–24); P2.5(CI90%):12(11−13); P97.5(CI90%):52(49–56).

ALT value for women (U/L): n= 1232; Minimum:4; Maxi-mum:41; Median(CI90%):16(16–17); P2.5(CI90%):9(8–9); P97.5(CI90%):33(32–35).

AST value for men (U/L): n= 375; Minimum:11; Maximum:56;Median(CI90%):24(23–25); P2.5(CI90%):16(14–17); P97.5(CI90%):47(42–52).

AST value for women (U/L): n = 509; Minimum:10; Maxi-mum:35; Median(CI90%):20(19–20); P2.5(CI90%):14(13–14); P97.5(CI90%):30(29–31).

We observed higher concentrations for men than for women,with an average difference of 8.5 U/L for ALT (p b .0001) and 5.9 U/Lfor AST (p b .0001).Therefore, RIs must be different for each gender.

ALT men:12-50 U/L; ALT women: 9-33 U/L; AST men 16-47 U/Land AST women: 14-30 U/L.

Conclusions

We have established our own RIs for ALT and AST through ourown analytical method and population.

As we can comprobe, these RIs are different between men andwomen, and are slightly different from those described by themedical technology company, especially for the AST. Therefore, ourlaboratory will proceed to its replacement.

doi:10.1016/j.cca.2019.03.766

T379

Alanine aminotransferase: Upper limit of normal must bequestioned

M. Guemra, P. Gabriel, Y. Villena, A. Blanco, A. Rando, R. LópezMartínez, F. Rodríguez FríasHospital Vall d'Hebron, Barcelona, Spain

Background-aim

The upper limit of normal (ULN) of alanine aminotransferase(ALT) is used as a discriminating value between liver injury andhealthy status. Significant differences in ULN among laboratorieshave been reported and experts opinion suggest that ALT ULN isoverestimated.The aim of this study is to verify if the discriminativevalue of ALT is lower than the ULN used in our laboratory (50 U/Land 35 for adult man and woman respectively), by comparing ALTvalues obtained in a cohort of hepatitis C virus (HCV) infectedpatients after reaching a sustained virological response (SVR) withdirect-acting antivirals (DAA).

Methods

HCV RNA (Cobas 6800, Roche), platelet count (XN, Sysmex) andALT and aspartate aminotransferase (AST) activities (AU5800Beckman-Coulter) were obtained from 137 men and 130 women(14 to 77 years of age) before and after reaching SVR. Patients withliver fibrosis were excluded according to Fibrosis Index (FIB-4) b1,45.


Kolmogorov-Smirnov (double-sided 95% interval) test was used toassess ALT normal distribution and 97,5th percentile (ULN) wascalculated in both groups. U-Mann-Whitney test was used todetermine differences in ALT ULN between men and women afterSVR.Wilcoxon test was performed on naïve-treatment patients withpositive HCV RNA (N15 IU/mL) and normal ALT, to compare ALTvalues before and after treatment.

Results

Normal distribution was rejected in both groups (p b 0,05). AfterSRV, ALT 97,5th percentile was 30 U/L for men and 25 U/L for women(U = 10,215; p b 0,0001). Using the ULN provided by the manufac-turer, 58,4% men and 41,5% women had normal ALT at pretreatment.In this subgroup, ALT 97,5th percentile went from 47 U/L (men) and35 U/L (women) to 35 U/L and 21 U/L respectively, after SVR (Z =5,57; p b 0,0001).

Conclusions

ALT ULN provided by the manufacturer suggested lack of liverinjury in up to 58,4% men and 41,5% women with HCV infection.Lower ALT ULN values obtained after SVR (30 U/L for men and 25 U/Lfor women) evidence liver injury despite “normal” ALT at pretreat-ment. Misclassified patients with “silent” liver injury was reduced to19,7% men and 15,3% women using ALT ULN found in this study.Finding healthy control population is currently the main issue in theintent of defining normality for ALT.

doi:10.1016/j.cca.2019.03.767

T380

Comparison of fecal calprotectin chemiluminiscence test with aELISA test

T. Alvarez Fernandez, L. Prados Lourdes, O. Segastagoia Epelde, A.I.Cillero Sanchez, I.P. Constanso Conde, A. Perez FuertesHospital Xeral Arquitecto Marcide, Ferrol, Spain

Background-aim

Calprotectin is the most abundant cytoplasmic protein ofneutrophils. Its concentration in human stool samples is significantlyhigher in infectious and inflammatory processes, and has become animportant tool for the assessment of inflammatory bowel disease(IBD). Thus, fecal calprotectin is used both as a diagnostic andfollow-up to this disease. This has led to an increased demand forfecal calprotectin in the laboratory and forced the replacement ofolder and slower ELISA assays with newer and automated such aschemiluminescence (CLIA) assays. The aim of the study was to provethe correlation of the fecal calprotectin results between a ELISA assayand a CLIA assay.

Methods

For 2 weeks, the fecal calprotectin samples were processed inparallel by the currently in use assay (ELISA, Bühlmann fCal) and bythe new incorporated assay (CLIA, QUANTA Flash Inova). Theircorresponding extraction kits were used. All samples were dupli-cated by both assays. We studied the correlation of quantitativeresults using a Bland-Altam plot and a Passing-Bablok regression.After that, the results were categorized according to the

manufacture's recomendations. ELISA: negative “-”(b50 mg/Kg),indeterminate “?” (50–200 mg/Kg), positive “+” (N200 mg/Kg).CLIA: negative “-” (b50 mg/Kg), indeterminate “?” (50-120 mg/Kg),positive “+” (N120 mg/Kg). To evaluate the agreement between “-”,“?” and “+” classes we used the Kappa statistic. When applicable, theresults are accompanied by the 95% Confidence intervale inparentheses. Statistical analyses were performed using MEDCALCsoftware.

Results

55 samples were processed, one of which was excluded becauseaberrant results. With the remaining 54 the regression andconcordance study. The Bland-Altam plot showed a proportionalerror of −79% (−3.3 to −154.8) of the CLIA assay versus ELISA assay.The Passing-Bablok regression curve was “y = 0,25x + 9,31” (y =“CLIA”, x =“ ELISA”), m = 0,25(0,20 to 0,34) and b = 9.31(5.48 to11.45). For this regression, Cusum test for linearity showedsignificant deviation from linearity (p b .01). The agreement betweencategories was moderate: Kappa = 0.45 (0.27 to 0.63). Results byclasses (the number before the method mean subjects): 21 ELISA “-”= 21 CLIA “-”; 19 ELISA “?”= 13 CLIA “-”+ 6 CLIA “?”; 14 ELISA“+”= 6 CLIA “?”+ 8 CLIA “+”.

Conclusions

There was no correlation between the quantitative results of bothassays so the results from the new CLIA assay could not be used forthe follow-up of patients with previous ELISA assay results. Theconcordance between assays to classifying patients is moderate, so itis mandatory to include the new reference intervals with clinicalsignificance in the laboratory reports.

doi:10.1016/j.cca.2019.03.768

T381

Does the environmental temperature influence all screeningprograms for colorectal cancer?

J. Wong-Artetab, A. Irurzunb, I. Bilbaoa, I. Idigorasa, I. PortilloaaColorectal Cancer Screening Programme Coordinating Centre, BasqueHealth Service Bizkaia, SpainbDepartment of Biochemistry, Donostia University Hospital, SanSebastián, Spain

Background-aim

In previous studies it has been described that the increase inenvironmental temperature decreases the values of hemoglobin infaeces (f-Hb) increasing the rate of false negatives. Each colorectalcancer (CRC) screening program uses a different model, so it isimportant to know how temperature can affect different models. Theobjective of this study is to see if the environmental temperatureaffects the result of f-Hb in our working model.

Methods

A retrospective study of all f-Hb results from the CRC screeningprogram obtained in two different areas of our population during theyears 2016 and 2017 was made: group 1 mean temperature 14.2 °C(−6.5 to 39 °C), group 2 mean temperature 12.8 °C (−2.9 to 37 °C).The average daily temperatures recorded on the day of the analysis


of the samples in the Laboratory and the average temperature duringthe summer and winter months were correlated with the results of f-Hb and the percentage of positivity of the fecal occult blood test(SOH) (cut-off point 100 μg Hb/L). The analyzer used was OC-SensorDIANA - Palex Medical.

Results

A total of 19,315 results were collected (group 1: 8423, group 2:10892). When correlating the months of highest average temper-ature (June–July, group 1: 22 °C, group 2: 19.8 °C) with the monthsof lowest temperature (November–December, group 1: 5.4 °C,group 2: 8.4 °C) it was observed that the values of f-Hb werehigher in the months of higher temperature (group 1: 31 vs 22 μgHb/L, group 2: 25.2 vs 18.1 μg Hb/L). In addition, when analyzingthe results of f-Hb in different cut-off points of medium temper-ature, it can be seen in group 1 that to higher temperature the f-Hbvalue increases (N 15 vs b15 °C: 26.2 and 25.3 μg Hb/L; N 20 vs b10°C: 29.4 and 22.1 μg Hb/L; N 25 vs b5 °C: 25.9 and 11.6 μg Hb/L);although in group 2 this pattern was not observed. The positive(SOH) test percentages in the different comparison groups did notshow important differences.

Conclusions

The results obtained in our study show that in CRC screeningmodels like ours, where the SOH kits are delivered by theparticipants in their primary care center and from there they aresent to the Laboratory and analyzed on the day of their arrival,increases in ambient temperature do not seem to increase the falsenegative rate. This study presents several biases such as notrecording the temperatures prior to the analysis of the kit in theLaboratory, nor the patient's round number; but it serves as a firststep of recruitment for a prospective study.

doi:10.1016/j.cca.2019.03.769

T382

Estimation of leptin, adiponectin and C peptide levels in patientsof non-alcoholic fatty liver disease (NAFLD)

S. Yadava, M. Aroraa, B. Kabia, R. Chawlab, N. NegibaDepartment of Biochemistry, VMMC & Safdarjung Hospital, New Delhi,IndiabDepartment of Medicine, VMMC & Safdarjung Hospital, New Delhi,India

Background-aim

Non-alcoholic fatty liver disease (NAFLD) is recognized as themost common type of chronic liver disease, is a prevalent conditionassociated with obesity and insulin resistance (IR). In insulinresitance C peptide levels are raised and that may be affected byleptin levels. Leptin, a 16-KD peptide hormone that is releasedpredominantly from adipocytes, has been shown to be involved inperipheral insulin resistance and has a potential dual action onNAFLD experimental models, exerting a possible anti-steatotic, butalso a proinflammatory and profibrogenic action. Adiponectin is themost abundant adipose-specific adipokine. There is evidence thatadiponectin decreases hepatic and systematic insulin resistance, andattenuates liver inflammation and fibrosis.

Methods

A case control study was conducted in 50 patients of non-alcoholic fatty liver disease (NAFLD) in the Deptt of biochemistry,VMMC and Safdarjung Hospital, New Delhi.

Serum Leptin, Adiponectin and C-peptide levels were measuredusing Elisa kit.

50 age and sex matched people were taken as controls.

Results

Serum leptin and c peptide levels were significantly higher incases as compared to controls (p valueb.005). Serum Adiponectinlevels were significantly higher in controls as compared to cases (pvalueb.005).

Conclusions

According to the results of the present study, leptin and c peptidelevels were raised and Adiponectin levels were lowered in casessignificantly as compared to the control group. Obesity has beenrecognized as a risk factor the development of chronic liver diseasecaused by a variety of aetiological factors including NAFLD. Thiscondition has been associated with high serum leptin and c peptidelevels. Adiponectin is an abundant adipocyte-derived hormone withwell-established anti-inflammatory and insulin sensitizing proper-ties. The significance of adiponectin in protecting obesity-relatedNAFLD has been increasingly recognized. Further well-controlledstudies in large number of patients are needed to elucidate whetherleptin, Adiponectin and c peptide have any diagnostic role in NAFLDpatients.

doi:10.1016/j.cca.2019.03.770

T383

Fecal calprotectin levels and degree of inflammation in inflam-matory bowel diseases patients

M. Yordanovac, D. Gerovad, A. Atanasovaa, B. Galunska-KalchevabaDepartment of Anatomy and Cell Biology, Faculty of Medicine, MedicalUniversity of Varna, BulgariabDepartment of Biochemistry, Molecular Medicine and Nutrigenomics,Faculty of Pharmacy, Medical University of Varna, BulgariacDepartment of General Medicine and Clinical Laboratory, Faculty ofMedicine, Medical University of Varna, BulgariadDepartment of General Medicine and Clinical Laboratory, Faculty ofMedicine, Medical University of Varna, Bulgaria

Background-aim

The inflammatory bowel disease (IBD) is a serious health problemwith chronic recurrent course and increasing frequency. IBD afectspredominantly young people with poor quality of life and isconsidered to be due to dietary disorders, stress, multiple medicationuse, poor oral hygiene, smoking and alcohol abuse. The quantitativedetermination of fecal calprotectin (FC) as a noninvasive diagnostictool to monitor the inflammation process is useful to differentiateorganic from functional bowel diseases and may reduce the need forunnecessary invasive procedures.

Aim: To assess the role of FC in evaluating inflammatory boweldisease activity and its correlation with the ESR and CRP.


Methods

Thirty IBD patients were enrolled in the study: seventeen patientswith ulcerative colitis (UC) and thirteen with Crohn's disease, agedfrom 21 to 70. Based on the endoscopic and clinical finding thepatients were divided into two groups. The first group comprised 20patients with severe form of the disease and the second – 10 patientswith moderate form of the disease or in remission. CRP and ESR weredetermined in blood as nonspecific markers of inflammation. FC as amarker of neutrophil flux to the intestine was measured by ELISAmethod.

Results

FC values in all tested patients vary from 134–1800 μg/g. In thefirst group the mean of FC values is 1492 ± 313 μg/g. In the secondgroup the levels of FC are significantly lower (mean 371 ± 168 μg/g; p b .01). The diagnostic sensitivity and specificity of FC is 82% and98% respectively based on the value of 350 ⎧g/g as a cutoff value.CRP values range from 13.9 to 90.4 mg/l in patients with severe formof the disease and were significantly higher when compared to thosein patients with mild forms of the disease or in remission (range 2.2–8.9 mg/l, p b .019). A good correlation is established between FC andCRP levels (r = 0.56) in the first group. In the same group the ESRvalues are increased too but the correlation with FC levels is weaker(r= 0.41). These relationships are less pronounced in the secondgroup.

Conclusions

FC has better diagnostic sensitivity and specificity than traditionalsystemic inflammatory markers and is a good additional diagnostictool in screening patients with IBD.

doi:10.1016/j.cca.2019.03.771

T384

PIIINP and TIMP1, reference values for children and young adults

S. Antonsena, B. Hoffmann-Petersenb, I. Thusingb, H.M. Rasmussenb,P. Wittenhagenb

aDepartment of Clinical Biochemistry, OUH Svendborg Hospital,Svendborg, DenmarkbH.C. Andersen Children's Hospital, Odense University Hospital, OdenseC, Denmark

Background-aim

The amino terminal peptide of type III procollagen (PIIINP) is aproduct of type III collagen turn over and is used clinically as abiomarker of fibrosis of the liver, kidney and other tissues. Tissueinhibitor of metalloproteinase 1 (TIMP1) is a circulating inhibitor ofmatrix metalloproteinases, which increases fibrosis. TIMP1 seems tobe an important player in fibrogenesis, although the exact role of thisinhibitor is still not fully elucidated. Little is known about the normallevels of these markers in childhood, where they are expected toreflect collagen production during normal growth.

Aim of this study was to investigate reference values for PIIINPand TIMP1 for children and young adults.

Methods

PIIINP and TIMP1 were measured by chemiluminescence immu-noassays, using two monoclonal antibodies on an ADVIA Centaur XP(Siemens Healthcare Diagnostics) in serum from 468 healthyindividuals (242 males and 226 females), aged 6–28 years.

Results

PIIINP is higher in young males compared to females. Levels arerelatively high during childhood with a marked increase inadolescence. PIIINP reference intervals: Males: 6–11,9 years: 13–30μg/L (N= 51); 12–14,9 years: 10–60 μg/L (N= 90); 15–16,9 years:10–34 μg/L (N= 28); 17–20,9 years: 7–24 μg/L (N= 39) and 21–27,9 years: 6–13 μg/L (N= 34). Females: 6–9,9 years: 13–25 μg/L (N= 31); 10–14,9 years: 13–50 μg/L (N= 53); 15–19,9 years: 6–22 μg/L (N= 67) and 20–27,9 years: 3–13 μg/L (N= 75). TIM-P1concentrations are higher in males compared to females, andconstant during the age interval investigated, while females show adecrease in concentrations around the age of 20 years: TIMP1reference intervals: Males 6–27.9 years: 153–284 μg/L (N= 242).Females: 6–19,9 years: 147–255 μg/L (N= 151) and 20–27,9 years:70–240 μg/L (N= 75).

Conclusions

PIIINP is relatively high during periods of growth i.e. in childhoodand especially during adolescence. We observed higher levels ofPIIINP in females than in males in the age group 20–27.9 years.TIMP1 is constant in males aged 6–28 years and higher than infemales. Concentrations of TIMP1in females decrease after the age of20.

doi:10.1016/j.cca.2019.03.772

T385

Fibroscan® and ELF score in the study of fibrosis in obese patientswith chronic liver disease: A preliminary study

M.M. Arrebola-Ramireza, M. Rodriguez-Espinosaa, A. Dayaldasani-Khialania, E. Rodriguez-Garcíaa, E. Morcillo-Jimenezb, M. Bocanegra-Viniegrab, J.M. Rodrigo-Lópezb, M. Jiménez-PérezbaClinical Laboratory, Hospital Regional Universitario de Málaga, SpainbDepartment of Gastroenterology, Hospital Regional Universitario deMálaga, Spain

Background-aim

The stage of fibrosis is the most important single predictor ofsignificant morbidity and mortality in chronic liver disease. For years,liver biopsy has been the gold standard to evaluate liver fibrosis.Currently, non-invasive alternatives are available to quantify fibrosis.Transient elastography or FibroScan® quantifies liver rigidity, whichis proportional to the grade of liver fibrosis. Nevertheless, FibroScan®has some limitations, such as liver stiffness measurement failure inpatients with narrow intercostal spaces or high body mass index.Another approach for evaluating liver fibrosis involves the use ofserologic markers (standard clinical chemistry markers and param-eters of matrix metabolism) and scoring systems. The Enhanced Liver


Fibrosis (ELF) score is a commercially available index based on serumconcentrations of hyaluronic acid, aminoterminal propeptide oftype III collagen and tissue inhibitor of metalloproteases-1. ELFscore has been shown to predict disease progression in severalclinical settings: nonalcoholic fatty liver disease, nonalcoholicsteatohepatitis, chronic hepatitis B and C, liver cirrhosis, systemicdiseases and others.

The aim of this study was to estimate the concordance betweenELF score and FibroScan® in obese patients with chronic liver diseasedue to HCV and without liver biopsy, and the agreement betweenthe ELF score with indirect indices (aspartate aminotransferase-to-platelet ratio index (APRI) and Forns index).

Methods

Thirty patients suffering chronic hepatitis C whose body massindex was N35 were studied. All of them underwent a FibroScan®,and direct (for ELF score) and indirect serological markers (platelets,cholesterol, GGT and AST) of the hepatic fibrosis process weredetermined. Indirect markers were used to calculate the APRI andForns indices. The cutoff points of the ELF values to establish thedifferent degrees of fibrosis (F0-F4) were taken from recent scientificpublications. The kappa coefficient was calculated to evaluate theagreement between ELF score and Fibroscan®, and between ELFscore and the different indices calculated.

Results

Three patients (10%) obtained an ELF® score under 7.7 (equiv-alent to fibrosis stage F0), but had no Fibroscan® results due to afailure in the measurement. Of the 11 patients (36.7%) who obtainedan ELF® score between 7.7 and 9.8 (equivalent to fibrosis stage F1-F2), eight had a Fibroscan® result δ F2, and three a Fibroscan® resultN F2. An ELF® score between 9.8 and 11.3 (equivalent to fibrosisstage F2-F3) was obtained in ten patients (33.3%), six of whom had aFibroscan® result ε F2 but b F4. The Fibroscan® results were notavailable in the other four. Six patients (20%) obtained an ELF® scoreN11.3 (equivalent to cirrhosis), of whom four had a Fibroscan® resultε F4, and the other two had no results.

Regarding the Kappa index between parameters, we observedgood agreement which was statistically significant between the ELF®score and the APRI index (K = 0.71, 95% CI = 0.32 to 1.1). Theagreement between the ELF® score and the Fibroscan result® wasmoderate but not statistically significant (K = 0.41, 95% CI =−0.47to 0.871), and between the ELF® score and Forns Index it was poorand also not statistically significant (K = −0.06, 95% CI = −0.6 to0.5).

Conclusions

Although the number of patients studied is still too low to drawdefinitive conclusions, it is evident that direct serological markerscan be useful in obese patients in whom a Fibroscan cannot beperformed, (in our study 30% of the patients), regardless of thepossible degree of liver fibrosis, as the results obtained by bothmethods coincided to a great extent.

Regarding the indices calculated with indirect markers, the onewith the greatest concordance with ELF is the APRI index.

doi:10.1016/j.cca.2019.03.773

T386

Macroamylasemia: Frequency of appearance and MOST affectedranges of age and sex

L. De La Hoz Gil, V. O'valle Aisa, N. Hernández Martínez, V. SeijasMartínez-EchevarríaBiochemistry Department, University Hospital Severo Ochoa, Leganés,Spain

Background-aim

This study aims to determine the frequency of appearance ofmacroamylasemia in our sanitary area from January 2017 toNovember 2018. Altered biochemical characters (amylase, lipaseand macroamylase) had been also analyzed, as well as the reason forthe request and most affected ranges of age and sex.

Methods

A cross-sectional study was carried out, making a research in theOMNIUN database. The technique used in this study had been theprecipitation with polyethylenglycol 6000 at 24%. A value higherthan 60% is considered pathological.

Results

There were 67 requests, of which 33 weren't determinedbecause 24 of them (72,8%) had amylase and lipase levels withinthe reference range, 7 (21,2%) showed an increase in amylase andlipase levels (acute pancreatitis) and 2 (6%) had previouslybeen diagnosed with macroamylasemia. The remaining 34 testswere determined, concluding that 10 (29,4%) of them werepathological.

Out of the total of 67 tests, 33 belonged to women (49,3%) and34 to men (50,7%). In adittion, the age ranges were: 4 (5,8%)patients between 0 and 15 years old, 6 (8,9%) between 15 and 30years old, 35 (52,9%) between 30 and 65 years old and 22 (32,4%)over 65 years old. Out of the 12 pathological macroamylasemias, 10belonged to women (83,33%) and 2 to men (16,67%). Likewise, theage ranges in this case were: 1 (8,33%) patients between 0 and 15years old, 6 (50%) between 30 and 65 years old and 5 (41,67%) over65 years old.

Regarding the reason for the request, 8 (66,8%) of the patholog-ical tests were cases of persistent amylase increase without anyassociated disease. In the rest of the 4 tests, the amylase increase wasassociated to Chron disease, B12 vitamine defficiency, celiac diseaseor ictus (in proportions of 8,3% respectively).

Conclusions

In conclusion, this study shows that the frequency ofmacroamylasemia in female population is 4 times higher than inmale population, especially in ages between 30 and 65 years old.Moreover, the amylase increase is the most common reason for therequest. At last, it should be clarified that the diagnostic criteriaaren't yet clear because 33 of the 67 tests weren't determined.

doi:10.1016/j.cca.2019.03.774


T387

Evaluation of human pancreatic elastase-1 quantification infaeces extracted with a conventional method (weighing) andwith the CALEX® Cap using the ScheBo® ELISA-KIT

A. Pomazanovsb, A. Röselc, H. Hartunga, C. Niederbergera, R. MauererdaBÜHLMANN Laboratories AG, Schönenbuch, SwitzerlandbDivison of Immunology I, Medical Laboratory Synlab, Weiden, GermanycHead of Division of Immunology I, Medical Laboratory Synlab, Weiden,GermanydMedical Director, Medical Laboratory Synlab, Weiden, Germany

Background-aim

Human Pancreatic Elastase-1 (E1) is an important markerreflecting exocrine pancreatic function. E1 has to be extracted fromfecal specimens with a complex and time-intensive extraction, theautomation of extraction is needed. In our laboratory we haveevaluated and compared the automatable CALEX® Cap extractiondevice to the conventional extraction method. 176 stool sampleswere collected and extracted with both methods for quantitative E1-determination and comparison. The correlation was good: r= 0,846,and the concordance of diagnostic value among all patients was 90%.In this work we showed that both extraction methods may be usedinterchangeably. This makes the full-automated E1 quantification instool using the CALEX® Cap extraction possible.

Methods

176 stool samples were collected with a stool specimen collector.Samples was homogenized, portioned and extracted with bothmethods for comparison. The extracts from both methods weremeasured with the ScheBo® Pancreatic Elastase 1™ Testkit. For aPassing-Bablok regression and Bland-Altman-Plot we have selectedonly the values within the measuring range (15–500 μg/g) resultingin a total of n = 58.

Results

The correlation of the two extraction methods (Fig. 1., Passing-Bablok regression, n = 58). The fit was good: r = 0,846 despite somescatter due to manual sample preparation. The concordance ofdiagnostic value among all patients shown in Fig. 2 was 90% (158/176). E1 b 100 μg/g indicates an advanced, E1 100–200 μg/g indicatesa mild pancreatic insufficiency and the E1 N 200 indicates theabsence of pancreatic insufficiency.

The 95% limits of agreement (−146,0; 139,4; Bland-Altman-Plot,Fig. 3) contain 95% (55/58) of the difference scores. The meandifference (bias) of the measurements (−3,28) betweenCALEX® Cap with ScheBo® ELISA and weighing with ScheBo® ELISAdo not exceed the maximum allowed difference between themethods. Both methods are in agreement and may be usedinterchangeably.

Conclusions

In this work we showed that both extraction methods (CALEX®Cap extraction and conventional method) are in agreement and maybe used interchangeably. This makes the full-automated E1 quanti-fication in stool using the CALEX® Cap extraction possible. Simpleand reliable sample preparation of stool specimens significantlylowers the hands-on time for fecal samples. The CALEX® Cap

extraction is not only suitable for the E1 determination, but for thefecal calprotectin as well.

doi:10.1016/j.cca.2019.03.775

^T388

Assessment of hepatic steatosis algorithms in non-alcoholic fattyliver disease

N. Eremic KojicClinical Centre of Vojvodina, Centre for Laboratory Medicine, Faculty ofMedicine, University of Novi Sad, Serbia

Background-aim

In order to optimize identification of persons with non-alcoholicfatty liver disease (NAFLD) several algorithms for hepatic steatosisweredeveloped. These available algorithms as well as an algorithm derivedusing biochemical and anthropometric data of our participants.

Methods

There were 77 participants with abdominal obesity: 43 withNAFLD and 33 without NAFLD. Body mass index (BMI), waistcircumference (WC) and hip circumference (HC), systolic anddiastolic blood pressure were assessed. Fibrinogen, high sensitiveC-reactive protein (hsCRP), aspartate aminotransferase (AST), ala-nine transaminase (ALT), gamma-glutamyl transferase (GGT), uricacid, ferritin, glucose, insulin, homocysteine, lipid status parameters,apolipoprotein A-I, apolipoprotein B and Lp(a)-lipoprotein weremeasured. Fatty liver was assessed by ultrasound with the presenceor absence of hepatic steatosis. Discovering the most significantfactor on the presence of NAFLD is assessed through logisticregression modeling. The predictor variables were chosen accordingto an algorithm derived from conducted factor analysis and otheravailable algorithms for hepatic steatosis.

Results

Participants with NAFLD had significantly higher BMI (34.38 ±9.73 vs 28.05 ± 4.79 kg/m2, p= .001), WC (108.05 ± 11.47 vs 96.15± 14.27 cm, p = .001), HC (114.93 ± 11.01 vs 108.21 ± 9.82 cm, p= .050), systolic (128.98 ± 8.67 vs 122.42 ± 10.62 mmHg, p= .010)and diastolic blood pressure (83.64 ± 5.94 vs 78.33 ± 7.57 mmHg, p= .001), AST (23.93 ± 6.91 vs 21.70 ± 5.21 U/L, p= .014), ALT(30.50 ± 13.70 vs 23.00 ± 11.75 U/L, p= .007), hsCRP (4.34. ± 5.56vs 2.98 ± 2.34 mg/l, p = .004) and uric acid (358.02 ± 83.29 vs296.78 ± 84.54 μmol/l, p = .001), in comparison non NAFLD. Logisticregression model with algorithm derived from factor analysisshowed the best performance. From other available algorithms, onlyfatty liver index (FLI) and hepatic steatosis index (HSI) hadstatistically significant discriminatory power.

Conclusions

Elevation of WC, HC, BMI, DBP, SBP, Fbg, hsCRP, glucose and uricacid, incorporated in our hepatic steatosis prediction model, had thebest predictive power among all assessed algorithms.

doi:10.1016/j.cca.2019.03.776


T389

Impact of vitamin B6 deficiency on aminotransferase assays andclinical implications

J. Pekara, A. Reguemea, G. Lassaillyb, F. Loiseleura, I. Kimc, M. JoncquelChevalier Curtc, P. Maboudoua, G. Grzychc

aUniversity of Lille, CHU, Biochemistry Emergency Department, Labora-tory of Proteins Biochemistry, Lille, FrancebUniversity of Lille, CHU, Department of Digestive System Diseases, Lille,FrancecUniversity of Lille, CHU, Laboratory of Endocrinology, Metabolism-Nutrition, Oncology, Lille, France

Background-aim

Plasma transaminases activity can be measured by methods usingor not pyridoxal-5′-phosphate (PLP), active form of vitamin B6 andessential cofactor of these enzymes. The recommended methodrequires PLP addition but numerous laboratories use assay withoutPLP. The aim of this study are to evaluate the analytical relevance ofthe addition of PLP in the reaction medium according to vitamin B6levels and evaluate the clinical relevance of the lack of PLP to assaytransaminases activity by using Actitest®.

Methods

The impact of PLP addition on transaminases activity wasevaluated on 1086 plasma samples with known vitamin B6 plasmalevel from hospitalized patients (Lille hospital, Lille, France). Weevaluated the relative difference of catalytic activity of transaminasesaccording to methods measurement and plasma vitamin B6 level.Influence of on Actitest® scores was evaluated by same methodologyon 20 tests.

Results

Results confirms transaminases activity is higher for method withPLP (p-value b.0001) and a correlation between vitamin B6 levelsand the bias of the increase of catalytic activity of transaminases isdemonstrated when adding PLP in reaction medium (p-value b.0001for AST and p-value b.018 for ALT). Absence of PLP when measuringALT leads to decrease Actitest® score and is responsible of changesetgrade for 10 on 20 patients.

Conclusions

For aminotransferase measurements, the method recommendedby IFCC, with PLP, should be systematically used in biochemistrylaboratories to overcome possible vitamin B6 deficiencies andprevent physiological variations.

doi:10.1016/j.cca.2019.03.777

T390

Early prevention program for colon and rectal cancer. Dataanalysis

R. Jáñez Carrera, N.L. Hernandez Martinez, V. O'valle Aisa, L. De LaHoz GilBiochemistry Department, University Hospital Severo Ochoa, Leganés,Spain

Background-aim

Colorectal cancer (CRC) is the second leading cause of cancerdeath in our country and the most frequent malignant tumor whenboth sexes are considered together. Most CRCs originate fromcolorectal polyps, which are usually sporadic and correspond toadenomas.

The CRC meets the criteria to justify its early detection as it isserious, frequent, preventable and there is a non-invasive screeningtest consisting of the detection of the concentration of humanhemoglobin in faeces.

Since 2015, our area has implemented a programme of earlyprevention of colon and rectal cancer aimed at citizens between 50and 69 years of age, excluding those with high risk criteria. SinceJune 2017 our Hospital has joined this programme.

To carry out the analysis of the results obtained from thetest of hidden haemorrhages in immunological faeces, from ourincorporation in the early prevention programme of colon and rectalcancer.

Methods

Based on the results stored in the laboratory's computer system(Infinity de Roche) and its stadistic programme (Omnium), thenumber of samples and the number of positive tests have beenanalyzed for one year, with a concentration of human hemoglobin infaeces N100 ng/mL being considered a positive result.

The samples were processed in the Biogen OC-Sensor IOequipment, through an immunochemical latex agglutination test,which has the advantage of being specific for human hemoglobinwithout the need to previously restrict the diet.

Results

In our laboratory, 7434 samples were processed, of which 3374(45%) belonged to men with an average age of 63.5 and 4060 (55%)belonged to women with an average age of 63.2 years. Of thesesamples, 630 (8.5%) had a positive result, 378 (60%) belonged to menwith an average age of 65.1 years and 252 (40%) belonged to womenwith an average age of 64.3 years. Participants had an age range of 50to 70 years (fashion: 69 years).

Conclusions

According to our results, the percentage of positives (8.5%) ishigher than expected by the early prevention program for colon andrectal cancer, which was 7%. This we believe may be due to the highage of the participants (fashion: 69 years).

Although the percentage of participation has been higher inwomen, the percentage of positives has been higher in men.

doi:10.1016/j.cca.2019.03.778

T391

The alcohol dehydrogenase (ADH) and its isoenzymes as candi-dates for liver fibrosis in patients with hepatitis C

W. Jelskia, K. Orywala, T. Lapinskib, B. Mroczkob,caDepartment of Biochemical Diagnostics, Medical University, Bialystok,PolandbDepartment of Infectious Diseases and Hepatology, Medical University,Bialystok, Poland


cDepartment of Neurodegeneration Diagnostics, Medical University,Bialystok, Poland

Background-aim

The best known causes of liver fibrosis are chronic HCV, HBVinfections and alcohol abuse. Currently severity of liver diseasesare diagnosed through different methods e.g. blood tests such as:bilirubin, albumin, alanine aminotransferase. The “gold standard”for the diagnosis and assessment of liver disease is still liverbiopsy, but this exam has a number of limitations and disadvan-tages. Nowadays, noninvasive tests hold a promise for improveddiagnosis of liver fibrosis. In our previous study, we observed thatthe total alcohol dehydrogenase (ADH) activity was elevated inthe sera of patients with HCV infections. The increase in total ADHactivity was correlated with ADH class I and II and appears to bedue to the release of these isozymes from inflammatory cells ofthe liver.

Methods

Serum samples were taken from 74 patients (46 males, 26females, 30–74 years) suffering from viral hepatitis C and from 80persons of control. Class I and II ADH isoenzymes were measured byflurometric method using the specific substrates (4-metoxy-1-naphtaldehyde and 6-metoxy-2-naphtaldehyde). The activities ofclass III and IV were measured by photometric method with specificsubstrates. Total ADH activity was estimated by the photometricmethod.

Results

The total activity of ADH was significantly higher in patients withhepatitis C than in healthy (about 53%). The total activity of ADH was1165 mU/l in patients, and 540 mU/l in controls. The comparison ofADH isoenzymes activities showed that the high difference wasexhibited by class I and II ADH. The activity of these classesisoenzymes in the hepatitis C group increased respectively about57% (4.15 vs 1.76 mU/l) and 46% (25.26 vs 13.72 mU/l) in thecomparison to the control. The diagnostic sensitivity for ADH I was72.5%, specificity 74.1%, predictive values of positive and negativeresults were 79.4 and 75.6% respectively. The sensitivity for ADH IIwas 66.4%, specificity 69.5%, predictive values of positive andnegative results were 71,2 and 68% respectively. The diagnosticcriteria for total ADH was lower than for ADH I and ADH II. Areaunder ROC curve for ADH I and ADH II were 0.745 and 0,660respectively.

Conclusions

These results suggest a potential role for ADH (especiallyADH I and ADH II) as markers of liver fibrosis in patients withhepatitis C.

doi:10.1016/j.cca.2019.03.779

T392

Shortening lactose tolerance test: audacity or efficiency?

M. Munoz Calero, E. Salas Herrero, C. Montilla LopezHospital San Juan de Dios del Aljarafe, Bormujos, Sevilla, Spain

Background-aim

Despite the fact that the most sensible test for diagnosis of lactoseintolerance is the Lactose breath hydrogen Test, still many hospitalsperform Lactose Tolerance Test (LTT) that consists in oral adminis-tration of Lactose and monitoring glucose (GLU) at 0, 30 and 60 min(LTT 60). Failure of blood GLU to rise by ε20 mg/dL above the basallevel is diagnostic of abnormal LTT.

Nowadays LTT is a high demanded test in our health area andimplies a challenge due to its high demand. Recently some authorsobserved that duration of LTT could be shortened without affectingaccuracy in only two points (Basal and 30 min, LTT 30). Simplify thetest could help to reduce the waiting list we currently have becausetime expended for the test is lessened.

Methods

A retrospective observational study of LTT realized in laboratoryfrom 2013 to 2016 is performed. Patients were subjected to anoverload of 50 g of oral lactose. Blood GLU was quantified in fastingstate (Basal), at 30 and 60 min. GLU was determined in a DimensionVista 1500 (Siemens Healthcare). Increases of GLU lower than 20mg/dL were reported as abnormal LTT.

The objective of the study was to evaluate the closeness ofagreement between traditional LTT (LTT 60, 3 points) and ashortened test (LTT 30, 2 points). For this purpose the Kappa Indexwas performed (SPSS statistical program).

Results

The study involves 1545 patients who were submitted to a LTT.With classical LTT, 815 patients (53%) had an abnormal LTT whileLTT 30 had 829 (53%) abnormal results. Shortened LTT modified theinterpretation in 14 patients (0.8%) where the levels of GLUincreased at 60 min.

The results of Kappa Index (0.982, p b .001) demonstrated a goodconcordance between LTT 60 and LTT 30.

Conclusions

Abbreviate LTT does not affect to diagnostic accuracy of LTT andsupposes advantages. It involves improvement in patient comfortsince a third extraction is not necessary and less time is lost for thedevelopment of the test. Shortening the LTT also carries cost savingsbecause a third sample processing is not necessary and saves time ofphlebotomist staff in the context of optimize resources.

doi:10.1016/j.cca.2019.03.780

T393

Glycogenic hepatopathy: Infradiagnostic hepatic complication indiabetes mellitus type 1. Case report

M. Ruiz-Ginesd, J. Ruiz-Ginesc, D. Rodriguez-Gonzalezd, P. Maria-Piedadd, B. Moreno-Torresa, M. Agudo-Macazagad, J. Sastre-Marcosb,L. Maria-CarmendaDepartment of Anatomic Pathology, Hospital Virgen de la Salud, Toledo,SpainbDepartment of Endocrinology, Hospital Virgen de la Salud, Toledo,SpaincDepartment of Neurology, Hospital Virgen de la Salud, Toledo, Spain


dDepartment of Pathology and Laboratory Medicine, Hospital Virgen dela Salud, Toledo, Spain

Background-aim

Glycogenic hepatopathy (GH) is a rare complication of the poorlycontrolled type 1 diabetes mellitus (DM) characterized by thetransient liver dysfunction with elevated enzymes, hepatomegaly(without inflammation or fibrosis) and abdominal pain, caused by thereversible accumulation of excess glycogen in the hepatocytes. It israrely seen in patients with type 2 DM. An essential element in thepathophysiology of GH is the wide fluctuation in both glucose andinsulin levels, promoting the hepatic accumulation of glycogen bystimulation of glycogen phosphorylase and synthetase enzymes,resulting in hepatomegaly and elevated liver enzymes (histologicalfindings suggestive of apoptotic-necrosis: membrane damage). Theexact mechanism is unknown. The pancreatic enzymatic changesdepend on vasodilation of small hydrogen sulfide-dependent bloodvessels, which interferes with enzymatic resorption. Differential-diagnosis: hepatic steatosis (HS) and glycogen-storage-diseases(GSD). GH andHS are clinically indistinguishable. HS ismore prevalentin type 2 DM and can progress to advanced liver disease and cirrhosis.Dual gradient-echo-abdominal MRI can distinguish GH vs HS.

Methods

29-year-old woman with uncontrolled type 1 DM and recurrentdiabetic ketoacidosis (DKA) and pancreatitis (cholecystectomizedand empirical dilation of the sphincter of Oddi), assessed byepigastric abdominal pain, nausea, and vomiting.

Results

Laboratory evaluation showed metabolic acidosis, ketonuria, AST145 mU/mL, ALT 150 mU/mL, amylase 437 mU/mL and lipase 1112mU/mL. Extensive evaluation for causes of hepatitis (toxic, autoim-mune, genetic, and infectious diseases): negative. The radiologicstudy (ultrasonography/CT/dual gradient-echo MRI) revealed anenlarged liver with normal morphology. Liver biopsy: withoutreticulin fibrosis or inflammatory infiltrates. Swollen hepatocyteswith glycogen storage (PAS+). Diagnosis: GH.

Conclusions

Adequate glycemic control can result in complete remission ofclinical, laboratory and histological abnormalities. Awareness of thisentity, including hepatologists, is low. Clinicians should consider GHin patients with uncontrolled DM after ruling out other commoncauses. Although liver biopsy remains a gold standard, evaluationwith non-invasive tests may be considered as an alternative.

doi:10.1016/j.cca.2019.03.781

T394

Investigation of serum malondialdehyde in patients with chronicviral hepatitis

D. Terzievc, D. Terzievab, V. Andonovc, N. MatevaaaDepartment of Biostatistics and Medical Informatics, Faculty of PublicHealth, Medical University, Plovdiv, BulgariabDepartment of Clinical Laboratory, Faculty of Pharmacology, MedicalUniversity, Plovdiv, Bulgaria

cSecond Department of Internal Medicine, Gastroenterology Section,Faculty of Medicine, Medical University, Plovdiv, Bulgaria

Background-aim

Oxidative stress is one of the pathophysiological mechanisms inchronic liver diseases. Free radicals are highly reactive, causing lipidperoxidation and production of reactive aldehyde metabolites suchas malondialdehyde (MDA). Many patients with chronic viralhepatitis also have insulin resistance, which results in lipidmetabolism changes. The present study aims to investigate therelationship between serum MDA levels and some biochemicalfindings in patients with chronic viral hepatitis.

Methods

A total of 55 subjects were divided in two groups consisted of 26patients with chronic viral hepatitis and 29 healthy controls. SerumMDA levels were measured using Human malondialdehyde ELISA kit(MyBiosource, USA) and Sirio S microplate reader (SEAC, Italy).Relationship between MDA and fasting serum glucose, total choles-terol, HDL- cholesterol, LDL-cholesterol, triglycerides, insulin andHOMA-IR was investigated.

Results

Serum concentration of MDA was significantly increased inpatients with chronic viral hepatitis compared to controls (157.88± 10.31 vs 65.85 ± 3.22 nmol/ml, P b .0001). The correlation analy-sis showed a negative and nonsignificant correlation between MDAand total cholesterol (r =−0.053, P= .797), HDL- cholesterol (r =−0.243, P= .231), LDL- cholesterol (r=−0.088, P = .668), triglyc-erides (rho =− 0.037, P = .857), fasting glucose (rho = −0.093, P= .650) and HOMA-IR (rho =−0.155, P = .449) in chronic viralhepatitis patients. There is a positive and nonsignificant correlationbetween MDA and insulin (rho = 0.161, P = .404).

Conclusions

Increased MDA levels as a marker of lipid peroxidation suggest theexistence of oxidative stress in patientswith chronic viral hepatitis. Nocorrelation was found between MDA and studied biochemicalparameters. It is suitable to assess the correlation between MDA andother specific for insulin resistance metabolic parameters.

doi:10.1016/j.cca.2019.03.782

T395

Serum levels of glutathione peroxidase and superoxide dismutasein patients with chronic viral hepatitis

D. Terzievc, D. Terzievab, V. Andonovc, N. MatevaaaDepartment of Biostatistics and Medical Informatics, Faculty of PublicHealth, Medical University, Plovdiv, BulgariabDepartment of Clinical laboratory, Faculty of Pharmacology, MedicalUniversity, Plovdiv, BulgariacSecond Department of Internal Medicine, Gastroenterology Section,Faculty of Medicine, Medical University, Plovdiv, Bulgaria

Background-aim

Oxidative stress is defined as an imbalance between reactivespecies formation and reduced antioxidant defences. Many


antioxidants counterbalance the effect of oxidants, including super-oxide dismutase (SOD), glutathione peroxidase (GPx), catalase, etc.Oxidative stress contributes to diabetes, atherosclerosis, chronic liverdiseases. Recent evidence suggests that insulin resistance has a keyrole in the development of liver steatosis. The aim of our study is toevaluate the serum levels of SOD and GPx in patients whit chronicviral hepatitis.

Methods

The study included 26 patients with chronic viral hepatitis and 29healthy controls. Serum concentrations of SOD (Human Cu/ZnODPlatinum ELISA, Bender MedSystems, Austria) and GPx-1 (HumanGlutathione Peroxidase 1 ELISA, BioVendor, Czech Republic) weremeasured with a Sirio S microplate reader, SEAC, Italy. The fastingglucose, triglycerides, total cholesterol and HDL-cholesterol concen-trations were analyzed (Mindray BS 200e, China). Enzyme immuno-assay was used to measure serum insulin levels (NovaTec, Germany).Insulin resistance was calculated using HOMA method. Comparisonof quantitative variables between groups was performed by inde-pendent samples t-test or Mann-Whitney U test. P value b.05 wasconsidered significant.

Results

The patients with chronic viral hepatitis were significantly olderthan healthy controls (49.46 ± 3.07 vs 35.62 ± 2.42 years, P= .001).Patients have significantly lower serum HDL- cholesterol (P b .0001)and significantly higher triglycerides (P= .029), insulin (P = .001)and HOMA-IR (P = .001) compared to controls. There were nodifferences in glucose, total cholesterol and LDL-fraction levelsbetween two groups (P N .05). Our results show significant decreasedlevel of SOD (mean ± SEM) in chronic viral hepatitis patients ascompared to controls (25.46 ± 1.72 ng/ml vs 128.86 ± 22.84 ng/ml,P b .0001). There was no significant difference in the GPx-1 betweengroups (2.19 ± 0.58 ng/ml vs 1.38 ± 0.43 ng/ml, P= .344).

Conclusions

Our results indicate an induction of SOD in chronic viral hepatitispatients. This represents an oxidative stress existence, which plays arole in chronic viral hepatitis pathogenesis.

doi:10.1016/j.cca.2019.03.783

T396

Serum alt levels – A study in a local Singaporean population

C.P. Yeo, G. Goh, W.T.P. Cheong, S.Y.J. Wong, C.H.C. TanDepartment of Clinical Pathology, Singapore General Hospital, Singapore

Background-aim

Serum Alanine Transaminase (ALT) is commonly used for initialscreening or assessment of hepatocellular diseases. A 2017 practiceguideline by the American College of Gastroenterology (ACG)recommended that ALT upper reference limits (URLs) of 29–33 IU/L(males) and 19–25 IU/L (females) be used to prompt further clinicalevaluation. Clinical laboratorians have since expressed concerns overthe recommendations in view of differences in population andmeasurement methodologies. This paper aims to examine ALT levelsin our local population in relation to the ACG recommendations.

Methods

140 subjects (40 males, 100 females, 21 to 70 years old) from alocal population were studied. These subjects were trimmed from aninitial volunteer cohort with a lifestyle and medical historyquestionnaire and screening laboratory tests. Serum samples wereanalyzed on Beckman-Coulter AU5800 and Roche Cobas c702analysers, using reagents with and without pyridoxal-5-phosphate(P-5′-P). Data analysis was performed on the Analyse-IT software.

Results

ALT levels (2.5th to 97.5th percentile) from the Beckman-Coulteranalyzer were 14–65 U/L (males) and 7–49 U/L (females) whenmeasured with reagents using P-5′-P; and 12–55 U/L (males) and 6–41 U/L (females) when measured with reagents without P-5′-P. Onthe Roche analyzer, ALT levels were 14–63 U/L and 10–46 U/L formales and females respectively with reagents using P-5′-P; and 12–53 U/L and 7–37 U/L for males and females respectively withreagents without P-5′-P. Although the two analytical platformscompared well, Passing-Bablok ALT with P-5′-P Beckman-Coulter= 0.93 Roche +2.03; and ALT without P-5′-P Beckman-Coulter =0.97 (Roche) +0.62, paired t-test showed significant differences (p b

.0001) between results obtained with and without P-5′-P activationon both the Beckman Coulter and Roche analyzers.

Conclusions

ALT levels demonstrated in our study were higher than thoserecommended by ACG. Moreover, ALT levels obtained by reagentswith and without P-5′-P were significantly different. Our studyhighlight that considerations based on local populations and specificassay methodologies are important in elucidating clinically signifi-cant ALT levels.

doi:10.1016/j.cca.2019.03.784

T397

Helicobacter pylori infection in patients with hyperemesisgravidarum

M. Grozdovska Naumoskaa, V. Antovskab, J. LavcanskaaaPHI UC for Clinical Biochemistry, United StatesbPHI UC for Ginecology and Obstetrics, United States

Background-aim

One of the many problems during pregnancy is consideredhyperemesis gravidarum (HEG) whose aetiology is currently un-known. Helicobacter pylori infection is considered a possible riskfactor. For this purpose, we studied the incidence of Helicobacterpylori infection in patients with hyperemesis gravidarum (HEG).

Methods

The study included pregnant women (n = 121) aged 25–30 yearsin the first trimester of their pregnancy. The control group (n= 58)consisted of patients without hyperemesis gravidarum (HEG).Experimental group (n = 63) was composed of patients withhyperemesis gravidarum (HEG). Patients in the experimental groupwere asked about the exact time of the onset of symptoms, both ofwhich were monitored during pregnancy for symptoms and


outcome. The serum titer of IgG antibodies to Helicobacter pylori wasdetermined by ELISA technique.

Results

According to the findings of our study in the control group, 14pregnant women (24.14%) were positive for Helicobacter pylori. Inthe experimental group with hyperemesis gravidarum (HEG), 51pregnant women (80.95%) were positive for Helicobacter pylori,which compared with the control group showed a significantlysignificant difference (p b .001). Also, the mean serum IgG antibodytiter level for Helicobacter pylori is significantly higher in theexperimental group relative to the control (p b .001).

Conclusions

Because of the significantly significant difference between thetwo groups of subjects, the Helicobacter pylori infection may beconsidered a risk factor for hyperemesis gravidarum (HEG). Regard-ing the better management of Helicobacter pylori infections, werecommend that pregnant women who are pregnant before beingtested for Helicobacter pylori should be under control beforepregnancy.

doi:10.1016/j.cca.2019.03.785

T398

Biochemical non-invasive evaluation of nonalcoholic fatty liverdisease in a sample population

A. Guerra-Ruizb, A. Maiztegib, N. Fañanasb, S. Perezb, H. Sentissib, P.Iruzubietaa, J.C. GarridobaHepatology Unit, University Hospital Marques de Valdecilla, Santander,SpainbLaboratory Medicine, University Hospital Marques de Valdecilla,Santander, Spain

Background-aim

The incidence and prevalence of nonalcoholic fatty liver disease(NAFLD) are increasing and identification of people at risk of diseaseprogression is extremely important. Its importance rises as over-weight, obesity and diabetes arise and become pandemic. Nonalco-holic fatty liver disease (NAFLD) is a spectrum comprised of isolatedsteatosis, nonalcoholic steatohepatitis (NASH), advanced fibrosis,and cirrhosis. The risk for liver-related adverse outcomes (cirrhosisand mortality) increases significantly between those entities. Gener-ally, the diseases present silence, without clinical symptoms, whichmade it difficult to diagnose it at early stage, when interventionswould probably avoid main morbidities. Univocal diagnosis relies onliver biopsy which is very invasive and not extent of complications.Non-invasive diagnosis based on image and biochemical markers arebeing developed and currently is a dynamic field of research in orderto improve medical care of susceptible population.

We aim to evaluate the performance of different biochemicalNAFLD scores and to estimate the prevalence of this disease in ourpopulation.

Methods

We collected laboratory data from adult patients whose bloodsample arrive our Hospital Laboratory for routine analysis in the last

year. In those patients we estimate the risk and/or grade of NAFLD,NASH or fibrosis using NAFLD Fibrosis Score (NFS), FIB4, Fatty LiverIndex (FLI) and Hepamet Score. We exclude patients with diagnosedchronic or acute liver disease different from NAFLD, as well as thosewere some of the analytical or clinical parameters needed tocalculate the scores were not available. We evaluate the performanceof biochemical, non-invasive methods against transient elastographyusing controlled attenuated parameters (CAP), clinical diagnosis anddata from liver biopsy when available. Diagnostic accuracy wasassessed according to AUROC, sensitivity, specificity, positive andnegative predictive values, correct classification and grey zone.Prevalence of NAFLD was estimate using the biochemical score withbetter performance in this sample.

Results

We calculated NAFLD scores for 450 patients. Presence ofsignificant fatty liver disease (fibrosis N F1) vary between 15.6%and 67.1% according to the score used. Highest prevalence wasobserved when using NFS outcome. 62 of these subjects (14%) hadclinical records indicative of a diagnostic study in the specialist careservice, with 21 (5%) of them having underwent a diagnostic liverbiopsy. Diagnostic accuracy of Hepament Score was superior to NFS,FIB4 and FLI either using CAP (AUROC: 0.70, 95% CI 0.66–0.75) orLiver biopsy (AUROC: 0.75, 95% CI 0.69–0.80) as gold standard (p b

.05). There was not significant difference between NFS and FIB4scores and both were superior to FLI score (p b .05) estimating thepresence of significant fibrosis.

Prevalence of NAFLD in the population, using the Hepamet scorewas estimated at 26.4%. Around half of susceptible subjects (60 cases,51%) haven't had a record of specialist care service consultationregarding liver disease.

Conclusions

Non-invasive biochemical scores can be a very useful tools inscreening of fatty liver disease in general population, or at least in anopportunistic approach. Hepamet score showed superior perfor-mance than other NAFLD scores, improving diagnostic accuracy andcorrect classification. The prevalence of NAFLD is high, in line withcurrent reports, and it is important to start a methodic search forearly diagnosis in the susceptible population.

doi:10.1016/j.cca.2019.03.786

T399

Accuracy of new biomarkers for assessing liver fibrosis in chronicliver disease (CLD)

R. Loveroa, A. Schirinzia, A. Todeschinib, R. Rutaa, S. Marsicoa, A.Calamitaa, R. Gencoa, L. Varrasoa, A. Di Leob, F. Di SerioaaClinical Pathology Unit, University-Hospital Policlinico, Bari, ItalybGastroenterology Unit, University-Hospital Policlinico, Bari, Italy

Background-aim

Assessment and monitoring of liver fibrosis degree is importantfor CLD patient management. Transient elastography (TE), measuringliver elasticity, is considered a highly reproducible and reliableoption for grading liver fibrosis, except in some cases such as obesity.Although well-validated, TE is costly, limited to referral liver centers,and has a 5% to 10% failure rate. Blood biomarkers are therefore of


particular interest, as they can be applied in most clinical settings.The Enhanced Liver Fibrosis (ELF) test is a validated algorithmcombining 3 serum markers of extracellular matrix remodeling andfibrogenesis: hyaluronic acid, N-terminal pro-peptide of collagen III,and tissue inhibitor of metalloproteinase-1. Soluble isoform ofsuppression of tumorigenicity 2 (sST2) and Galectin-3 (Gal-3) haveemerged as biomarkers playing a prognostic role in cardiacremodeling and fibrosis. This study aimed to evaluate in CLD patientsthe usefulness of these biomarkers for assessing liver fibrosis statusand according to TE fibrosis grade.

Methods

We recruited serum from 42 CLD patients (28 males and 14females), with fibrosis-4 index assigned using FibroScan device(Echosens), and 40 samples from apparently healthy donors. ELFtest, Gal-3 were measured using chemiluminescent assay (AdviaCentaur XP and Architect i1000-Abbott respectively), and sST2 byELISA assay (DSX-Technogenetics). Diagnostic accuracy was testedusing ROC curves.

Results

ROC curves. ELF: AUC = 0.89 (95%CI: 0.78–0.95), p b .0001; cutoffN9.9 (sensitivity = 88%, specificity = 80%). Gal-3: AUC= 0.75 (95%CI: 0.62–0.85); p b .0001; cutoff N17.1 ng/ml (sensitivity = 72%,specificity = 80%). sST2: AUC = 0.82 (95%CI: 0.70–0.90) p b .0001;cutoff N59.9 ng/ml (sensitivity = 89%, specificity = 65%). ROC curvescomparison showed that association between ELF and Gal-3increases accuracy. Difference between areas: ELF-Gal-3: 0.139, p b

.005; ELF-sST2: 0.07, p= .2; Gal-3-sST2: 0.06, p = .3.

Conclusions

These preliminary data showed that ELF, Gal-3 and sST2 increasesignificantly in liver fibrosis. Our findings suggest that thesebiomarkers may support TE evaluation or represent a validalternative when this diagnostic system is not available.

doi:10.1016/j.cca.2019.03.787

T400

Serological diagnosis of celiac disease in pediatric patients: A stepforward usefulness of free light chain (FCLS)

V. O'valle Aísa, J. Jimenez Jimenez, N. Hernández Martínez, L. De LaHoz GilBiochemistry Department, University Hospital Severo Ochoa, Leganés,Spain

Background-aim

Celiac Disease (CD) is an immune-mediated systemic disorderelicited by gluten and related prolamines in genetically susceptibleindividuals. It's an enteropathy characterized by the presence of avariable combination of gluten-dependent clinical manifestations,CD-specific autoantibodies against tissue transglutaminase (tTGA),endomysial antibodies (EMA) and HLA haplotypes. The prevalence ofCD approaches 1% of the general population worldwide.

We aim to evaluate diagnosis of celiac disease in pediatricpatients, assessing serological markers and the usefulness of freelight chain (FCLs).

Methods

In this retrospective 5 years-term study, a total of 102 pediatricpatients, diagnosed with CD, showing serum anti-tTGA (N10 timesnormal values) and confirmed by positive EMA measurement, wereincluded for the analysis (44 boys and 58 girls: age range 1–16years). Genetic study for genotyping HLA and intestinal biopsy wasused to exclude CD or to stablish the diagnosis. Free light chain(FLCs) levels were assessed as marker of intestinal mucosa recovery.We used the Mann-Whitney U test for statistical analysis.

Results

All the individuals were IgA-competent. 32 (31,4%) bowelbiopsies were performed. Histological findings showed 78,2% villousatrophy Marsh stage 3 and in the remaining 21,8% Marsh stage 1,which were confirmed by genetic. Genetic testing was studied in 70patients: DQ2.5 (n = 63), DQ2.2 (n = 2), DQ2.5/DQ8 (n = 2), HLA-DQA1*05 (n = 3). At the time of diagnosis patients had low ferritinlevels (median 32,6 ng/ml) and FLCs (| + ⌊) was determined(median 42,2 mg/l); 26 patients were followed-up measuring FCLs.We observed a FCLs percentage decrease of 50,6% when tTGA levelswere normalizated (median FCLs 22,12 mg/l; p b 0,0001).

Conclusions

Decreasing trend in the incidence of biopsy-verified diagnoses isdue to new recommendations of ESGHAN guidelines, which celiacdisease diagnosis can be established based on serological and geneticmarkers. Normalization of free light chain levels as a potentialmarker of intestinal mucosa recovery can be useful to support thediagnosis and as predictive response biomarker.

doi:10.1016/j.cca.2019.03.788

T401

New cholinesterase assay for Thermo Scientific Indiko andKonelab clinical chemistry analyzers

N. Kivi, M. Kurki, M. Holopainen, M. Karppelin, S. Riistama-Laari, L.OtamaThermo Fisher Scientific, Vantaa, Finland

Background-aim

Acetylcholinesterase is an enzyme hydrolyzing acetylcholine.Cholinesterase levels in serum are used for the detection of patientswith atypical forms of the enzyme or as a test of liver function.Cholinesterase assay can be also used to detect possible insecticidepoisoning. A significant drop in serum enzyme activity is observedalready before the first symptoms and before neuromuscular effectsbecome apparent.

Methods

Cholinesterase assay is a two-reagent liquid test applied onThermo Scientific™ Indiko™ and Konelab™ clinical chemistryanalyzers from Thermo Fisher Scientific. Cholinesterase catalyzesthe hydrolysis of butyrylthiocholine to thiocholine and butyrate.Thiocholine reduces yellow potassium hexacyanoferrate (III) tocolorless potassium hexacyanoferrate (II). The decrease of absor-bance is measured at 405 nm.


Results

The assay measuring range is 400–16,000 U/l (Calibrator lot-dependent on Indiko™ platform) extended with automatic dilutionto 100–90,000 U/l. The repeatability (within-run precision) is 0.6–1.7% (CV; n = 80). Total precision (within analyzer) is 1.0–2.6% (CV;n = 80). Open on-board stability and calibration interval are 30 days.A comparison study was performed using commercially availablebutyrylthiocholine method as reference. Linear regression was y =1.076×–268.5 and r= 0.992 (n = 94).

Conclusions

This ready-to-use system reagent for Cholinesterase analysis onIndiko and Konelab analyzers is quick and accurate with excellentopen on-board stability.

doi:10.1016/j.cca.2019.03.789

T402

Stability of human hemoglobin in fecal samples at differentstorage conditions in fecal sampling device designed for colorec-tal cancer screening programs

C. Paparella, A. Barazzutti, A. Cugini, R. LuciniSentinel CH., Milan, Italy

Background-aim

Colorectal cancer screening programs have been established inmany countries in order to reduce incidence and mortality amongthe population. Collection of samples is generally done by theattendees at home and samples are returned to the laboratories forthe analysis. The use of a fecal sampling device able to stabilizehuman hemoglobin is therefore fundamental in various temperatureconditions.

Methods

The FOB Gold Tube Screen is a device optimized for the collectionand preservation of fecal samples. The buffer in this device is able toreduce the hemoglobin degradation at several temperature ranges. Inthis study, pooled human fecal samples have been used. The fecalsamples were collected using the collection device and stored atdifferent temperature ranges: -15/−25 °C, 2/8 °C, 18/23 °C, 24/27 °C,28/32 °C, 36/38 °C. The samples have been tested at regular intervalsup to 35 days at −15/−25 °C, 32 days at 2/8 °C, 17 days at 18/23 °C,14 days at 24/27 °C, 14 days at 28/32 °C, 7 days at 36/38 °C inorder to evaluate the recovery (% bias) versus the concentration atTime 0.

Results

After 35 days at −15/−25 °C, the recovery (% bias) vs Time 0concentration of hemoglobin in the pooled human fecal sampleswas 98.6% ± 9.7%. After 32 days at 2/8 °C, the recovery (% bias) vsTime 0 was 103.6% ± 11.4%. After 17 days at 18/23 °C, the recovery(% bias) vs Time 0 was 100.2% ± 20.1%. After 14 days at 24/27 °C,the recovery (% bias) vs Time 0 was 95.8% ± 17.4%. After 14 days at28/32 °C, the recovery (% bias) vs Time 0 was 92.1% ± 15.8%. After7 days at 36/38 °C, the recovery (% bias) vs Time 0 was 86.6% ±18.6%.

Conclusions

The use of FOB Gold Tube Screen allows to reduce the hemoglobindegradation in the fecal sample. Using this device, it's possible topreserve hemoglobin in the fecal sample by degradation for N30 dayswhen refrigerated, for 14 days when stored at temperature up to 30°C, and for 7 days when stored at temperature up to 38 °C. Theseresults confirm the ability of FOB Gold Tube Screen to reduce thehemoglobin degradation at various temperatures, making this devicefully suitable with the real condition that can be found in screeningprograms.

doi:10.1016/j.cca.2019.03.790

T403

Urinary orosomucoid: A new approach for the assessment ofCrohn's disease activity

B. Szirmayc, A. Tárnokd, P. Sarlósa, N. Szigetib, A. Ludányc, P. Kustánc,Z. Horváth-Szalaic, T. Kőszegica1st Department of Internal Medicine, University of Pécs Medical School,Hungaryb2nd Department of Internal Medicine and Nephrology Centre, Univer-sity of Pécs Medical School, HungarycDepartment of Laboratory Medicine, University of Pécs Medical School,HungarydDepartment of Pediatrics, University of Pécs Medical School, Hungary

Background-aim

Crohn's disease (CD), as one form of chronic inflammatory boweldiseases is characterized by alternation of relapse and remissionepisodes. To determine accurately the inflammatory activity of CD isstill challenging for gastroenterologists. Besides the generally usedclinical indices and endoscopic examination, laboratory markers alsohave a growing role in this process. The diagnostic benefit of urinaryproteins has not been well explored yet in relation to themanagement of CD. Our aim was to investigate whether urinaryconcentrations of orosomucoid can indicate the inflammatoryactivity of CD and to compare it with clinical scores and conventionallaboratory parameters.

Methods

Adult (n= 55) and pediatric (n= 31) patients with CD wererecruited and 68 healthy individuals (38 adults and 30 children)served as controls. Patients were classified by their activity indices(Harvey-Bradshaw Index (HBI) or Pediatric Crohn's Disease ActivityIndex (PCDAI)). Blood and urine samples of each participant wereanalyzed. Urinary orosomucoid (u-ORM) was determined by auto-mated immune turbidimetric assay and values were presented asurinary orosomucoid/urinary creatinine (u-ORM/u-CREAT) ratios inmg/mmol.

Results

U-ORM/u-CREAT ratios showed significant elevation both inchildren (0.50 vs. 0.07 mg/mmol, p b .001) and in adults with activeCD (0.32 vs. 0.14 mg/mmol, p= .01) compared with patients withinactive disease. Good correlation was observed between u-ORM/u-CREAT and the conventional inflammatory markers (hs-CRP, serumORM; p b .01) and clinical indices (HBI, p= .018; PCDAI, p b .001). U-


ORM/u-CREAT could discriminate between active and inactive CDwith similar performance as hs-CRP and serum ORM among pediatricpatients.

Conclusions

Based on our findings u-ORM/u-CREAT seems to be a valuableadditional marker in the assessment of CD activity, especially inchildren. The non-invasive simple sampling is a further advantage atthe outpatient service.

doi:10.1016/j.cca.2019.03.791

T404

Anti-kelch-like 12 autoantibodies in serum of patients withprimary biliary cholangitis

A. Bauera, T. Rawab, A. HabiorbaDepartment of Biochemistry and Molecular Biology, Centre of Post-graduate Medical Education, Warsaw, PolandbDepartment of Gastroenterology, Hepatology and Clinical Oncology,Centre of Postgraduate Medical Education, Warsaw, Poland

Background-aim

Primary biliary cholangitis (PBC) is a slowly progressing chole-static, autoimmune liver disease, characterized by the presence ofantimitochondrial (AMA) and antinuclear antibodies (ANA) in thepatients serum. PBC-specific ANA can be used to confirm thediagnosis of PBC, especially in AMA-negative cases. The newpotential autoantigen is the kelch-like 12 (KLHL12) protein. Theaim of the present study was to initially determine the autoanti-bodies reactive against this novel antigen - KLHL12 protein, in a wellcharacterized cohort of polish PBC patients. We evaluated also thecorrelation between level of anti-KLHL12, other specific antinuclearautoantibodies: anti-Sp100 and anti-gp210 and biochemical data.

Methods

Study population - 80 PBC patients, 30 pathological controls – 20primary sclerosing cholangitis (PSC) and 10 autoimmune hepatitis(AIH) patients, 15 healthy blood donors. AMA, anti-Sp100 and anti-gp210 were detected by commercially available kits (IMTEC-Human,Euroimmun; Germany and Inova Diagnostics; USA). The presence ofanti- KLHL12 antibody was determined by an” in-house” ELISAtechnique using a full-length recombinant human KLHL12 protein.

Results

Anti-KLHL12 antibodies were detected in 31% (25/80) of the totalcohort of PBC patients, including 25% (3/12) of AMA-negative PBC.They were found more frequently in PBC compared with non-PBCdisease controls (P b .001). We determined anti-KLHL12 in only oneof the 10 (10%) AIH patients. None of the sera of PSC patients orhealthy controls contained anti-KLHL antibodies. Specificity andpositive predictive values of this test for PBC were 98% and 96%,respectively. PBC sera contain antibodies which recognize variousnuclear protein. We found anti-KLHL12 together with anti-Sp100and anti-gp210 in 9 (36%) cases. Presence of anti-KLHL12 wasassociated with higher concentration of bilirubin.

Conclusions

Antibodies against KLHL12 protein are highly specific for PBC. Thecombination of anti-KLHL12 with other markers (AMA, gp210 andsp100) can increase the diagnostic sensitivity on PBC. They can aid inthe serologic diagnosis, also in cases in which AMA are notdetectable. Coexistence of different antibodies suggests an autoim-mune reaction against multiple nuclear components in some of PBCpatients.

doi:10.1016/j.cca.2019.03.792

T405

Extended range of the enzyme panel assays on the alinity Csystem using flex technology

M. Berman, L. RuvunaAbbott Diagnostics Division, Abbott Laboratories, Chicago, USA

Background-aim

Assessment of liver enzyme function is critical for monitoringdrug induced liver damage and detecting liver diseases such ashepatitis or cirrhosis. The presence in blood of liver enzymes such asAlanine Aminotransferase, is a useful indicator of enzymes leakingfrom damaged cells. This study established the assay specificcalibration factors for the enzyme assays from the Liver Panel usingknown traceable standards. The analytical performance of the LiverPanel was demonstrated on the Alinity c system using photometricand FLEX technology to establish an extended measuring interval forthe enzyme analytes.

Methods

Assay specific enzyme factors were established using increasingconcentrations of commercially available standards across thedynamic range on the Alinity c system. The FLEX methodology hasthe capability to extend the dynamic range of the assay without theneed for sample dilution and was demonstrated through keyperformance testing including linearity and method comparison.These studies were executed per Clinical and Laboratory StandardsInstitute (CLSI) protocols. The assay measuring interval wasestablished by the range for which acceptable performance for bias,imprecision, and linearity was met.

Results

The method comparison slopes for Amylase, Alanine Aminotrans-ferase, Aspartate Aminotransferase, Alkaline Phosphatase, Gamma-glutamyl transferase and Lactate Dehydrogenase ranged from 0.96 to1.03 with a correlation of 1.00. Known standards had slopes of 0.95to 1.01 with a bias b5% demonstrating strong correlation betweenARCHITECT and Alinity c systems. Performance studies demonstratedprecision b5% and samples were linear across the dynamic range ofthe assay. A reduction of retesting of high concentration samples dueto FLEX technology was observed for all assays. Significant reduc-tions of 24–73% were observed for the Amylase, Alkaline Phospha-tase, Alanine Aminotransferase and Aspartate AminotransferaseAlinity c assays.


Conclusions

Representative Liver Panel enzyme assays utilizing photometrictechnology on the Alinity c system demonstrated acceptableperformance with the on-market ARCHITECT clinical chemistryenzyme assays. The FLEX system seamlessly read high concentrationsamples in the upper dynamic range of the measuring interval afterinitial testing reducing the potential for delays in sample reportingdue to retesting or sample dilution.

doi:10.1016/j.cca.2019.03.793

T406

Distribution of lesions detected in a colorectal cancer screeningprogram

E. De Rafael Gonzalez, M.P. Picazo Perea, L. Rodelgo Jimenez, M.C.Lorenzo Lozano, M.S. Diaz Merino, A. Cabezas Martinez, M.A. RuizGinesComplejo Hospitalario de Toledo, Spain

Background-aim

In recent years many studies have demonstrated differences inclinicopathologic and prognosis features between tumors located inproximal colon (PC), distal colon (DC) and rectum (Re). This allowsto consider these three cancers as separate entities.

Proximal colon cancers are associated with subtle systemicsymptoms such as anaemia and asthenia and worse survival,whereas tumors located in distal colon or rectum present local signslike obstruction and present higher overall metastasis or recurrencerates after surgery. Location is relevant for therapeutic decision-making too.

The aim of this descriptive retrospective study is to evaluate thedistribution of lesions like colorectal cancer (CRC) and adenomatouspolyps, considered precancerous lesions, in an asymptomaticpopulation.

Methods

This study includes 1960 healthy patients, without any colorectaldisease, between 50 and 69 years old, who participate for a year in avoluntary colorectal cancer screening program with biannual fecaloccult blood tests (FOBT) (OC-Sensor Biogen Diagnostica®) followedby colonoscopy when results were higher than cut-point (100 μg/lhemoglobin) and polypectomy.

Results

187 patients (9.54%) had a positive result for FOBT in thescreening program.

17 CRC were detected: 9 (52.94%) were situated in DC, 6 (35.29%)in Re and 2 (11.76%) in PC.

Adenomatous polyps were biopsied in 112 patients. 64 patients(57.14%) had them in only one region: 36 were located in DC, 19 inPC and 9 in Re. While 48 patients (42.85%) had them in differentlocations: 32 patients in DC and PC, 11 in DC and Re, 3 in PC and Reand 2 patients had it across all the colon and rectum.

Conclusions

Attending the results, in this group of asymptomatic patients,adenomatous polyps and CRC have been found across all the colonand rectum, being distal colon the location most frequent, so to carryout a proper screening program, all the colon must be reviewedduring the colonoscopy.

Taking into account the differences in clinicopathologic andprognosis founded in the literature in colorectal cancer dependingon it's location, more studies about this distribution could allowfurther comparisons between symptomatic and asymptomaticpatients and improve colorectal screening programs.

doi:10.1016/j.cca.2019.03.794

T407

Role of an increase dietary fructose concentration in early processof cholelithiasis

R. Del Pozo, L. Mardones, M. Villagran, K. Muñoz, C. Cabezas, L.Troncoso, M. Mellado, M. MuñozDepartment of Biological Science, Faculty of Medicine, UniversidadCatolica de la Santisima Concepcion, Concepcion, Chile

Background-aim

Dietary fructose stimulates lipogenesis causing hepatic steatosis,but few information is available about its effect on biliary lipids.Biliary cholesterol is transported mainly by vesicles and micelles. Thefirst stage in the formation of gallstones corresponds to biliarycholesterol crystallization, derived from the vesicular transporters.The aim of this study was to investigate the influence of consuminghigh-fructose diet on serum lipids, and determine its effect ingallstones formation.

Methods

The experimental design was quantitative, and we had 2 groupsof BALB/c mice: one control (n= 20), and the other (n= 15) treatedwith increasing fructose concentrations (10%, 30%, 50% and 70%fructose in drinking water). After 1 or 2 months, the animals weresacrificed, and blood and bile samples were obtained. We deter-mined serum glucose and the corresponding lipid profiles. In bilesamples, cholesterol and phospholipids levels were analyzed, andcholesterol transporters (vesicles and micelles) were separated bygel filtration chromatography.

Results

Treated (2 months) animals showed: 1) no change in body weight(control: 31.4 ± 1.4 g vs. treated: 32.5 ± 2.6 g); 2) increase inglycemia (control: 90 ± 8 mg/dL vs. treated: 135 ± 18 mg/dL; p b

.001); 3) increase in serum triglycerides (control: 70 ± 36 mg/dL vs.treated: 362 ± 77 mg/dL; p b .05); 4) no change in serum totalcholesterol (control: 81 ± 11 mg/dL vs. treated: 91 ± 26 mg/dL); 5)no change in HDL-cholesterol and LDL-cholesterol; 6) slight increasein biliary cholesterol (cholesterol control: 3.6 mM, cholesterol treated:5.6 mM); 7) no change in vesicular and micellar phospholipids.


Conclusions

A high-fructose diet increase serum triglycerides and glycemia.But we did not observe significant changes in either biliary lipidconcentrations or biliary cholesterol transporters. We conclude thatfructose apparently does not alter the gallstone formation process inour experimental model.

doi:10.1016/j.cca.2019.03.795

T408

Effect of tacrolimus on serum low-density lipoprotein cholesterollevels in liver transplant patients

J. Djordjevicd, M. Culafica,b,c, S. Stankovicd, M. Kovacevica, S. VezmarKovacevica, B. Oluice, M. Stulicb,c, B. Miljkovica, D. Culaficb,caDepartment of Pharmacokinetics and Clinical Pharmacy, Faculty ofPharmacy, University of Belgrade, SerbiabClinic for Gastroenterology and Hepatology, Clinical Center of Serbia,Belgrade, SerbiacSchool of Medicine, University of Belgrade, SerbiadDepartment of Biochemistry, Clinical Center of Serbia, Belgrade, SerbiaeEmergency Center, Clinical Center of Serbia, Belgrade, Serbia

Background-aim

Lipid abnormalities have been described frequently in livertransplant patients. Variety of immunosuppressive medications actas a major factor influencing posttransplant lipidemic profiles.Tacrolimus (TAC) has been accepted as first-line treatment afterliver transplantation.

We aimed to evaluate the effect of TAC on low-density lipoprotein(LDL) cholesterol concentration in liver transplant recipients.

Methods

Patients regularly monitored at Clinic for Gastroenterology andHepatology following liver transplantation were prospectively en-rolled during the study period from January 2016 to July 2018.

Results

We included twenty-nine liver transplant recipients (male: 21),mean age 39.6 ± 13.7 on TAC with Mycophenolate mofetil andcorticosteroid therapy, in the study. Linear regression analysisrevealed following equation: Concentration (LDL) = 0.228 x Dose(TAC) + 2.966. Beta coefficient was estimated to 0.228 (95% CI0.120–0.337). Based on this calculation, a dose increase of 1 mg TACled to an elevation in LDL cholesterol concentration of 0.23 mmol/L.The regression model was statistically significant (p b .05) and 36.6%(R2) of variability was described by TAC dose. There was statisticallysignificant difference (p b .001) in LDL cholesterol depending onposttransplant period: the highest LDL cholesterol levels wererecorded in the first three months after transplant, the lowest after12 months of transplant (4.42 ± 0.54 mmol/L, 2.39 ± 0.44 mmol/L,respectively), but no significant decline was noted during 6–9months posttransplant.

Conclusions

Closer monitoring of lipid profile, LDL cholesterol concentrationsin particular, should be exercised with each increase of TAC dose,especially during the early posttransplant period.

doi:10.1016/j.cca.2019.03.796

T409

New amylase assay for thermo scientific indiko and konelabclinical chemistry analyzers

M. Kurki, N. Kivi, L. Virtanen, L. OtamaThermo Fisher Scientific, Vantaa, Finland

Background-aim

⟨-Amylase present in normal serum and urine is predominantly ofpancreatic and salivary gland origin. The enzyme catalyzes thehydrolysis of 1,4-⟨-glucosidic linkages in polyglucan chain to producesmaller units like maltose. ⟨-Amylase in serum and urine is primarilymeasured in the diagnosis of diseases of the pancreas. An increase inserum amylase activity may also be due to some other causes thanpancreatitis, like other pancreatic diseases, other acute intra-abdominal disorders, genitourinary diseases (e.g. renal insufficiency)or many miscellaneous causes (e.g. salivary gland lesions, diabeticketoacidosis and macroamylasemia).

Methods

Amylase assay is a two-reagent liquid test applied on ThermoScientific™ Indiko™ and Konelab™ clinical chemistry analyzers fromThermo Fisher Scientific. This reagent uses 4,6-ethylidene-(G7)-p-nitrophenyl-(G1)-⟨-D-maltoheptaoside (EPS-G7) as the substrateand is based on the recommendation of the IFCC. Once the substratehas been cleaved by ⟨-amylase, the produced smaller fragments canbe hydrolyzed by ⟨-glucosidase, which causes the release of thechromophore. The rate of increase in absorbance at 405 nm as PNP isproduced represents the total (pancreatic and salivary) ⟨-amylaseactivity in the sample.

Results

The assay measuring range is 10–1500 U/l extended withautomatic dilution to 10–7500 U/l. The repeatability (within-runprecision) is for plasma 0.5–1.3% (CV; n= 80), and for urine 0.6–1.0% (CV; n = 80). The within device (total) precision is forplasma 1.9–2.8% (CV; n = 80) and for urine 1.5–3.7% (CV; n =80). Open on-board stability and calibration interval are 30 days.A comparison study was performed using previous Amylasemethod (1-reagent liquid) as reference. Linear regression withIndiko was for plasma y (new) = 1.003 x (previous) –1.93, r =0.9995 (n= 89), and for urine y (new) = 1.033× (previous) –0.05, r= 0.998 (n= 63). Linear regression with Konelab was forplasma y (new) = 1.016 x (previous) –4.55, r = 0.9993 (n = 87),and for urine y (new) = 1.032× (previous) +0.22, r = 0.999 (n= 63).


Conclusions

With this ready-to-use system reagent, amylase analysis onIndiko and Konelab analyzers is quick and accurate with excellentopen on-board stability.

doi:10.1016/j.cca.2019.03.797

T410

The diagnostic accuracy of hematological laboratory parametersin the diagnosis of acute appendicitis in the adult emergencydepartment population

V. Radišić Biljaka, B. Bakulab, I. Soldob, Ž. Rašićb, A. ŠimundićaaDepartment of Medical Laboratory Diagnostics, University Hospital„Sveti Duh“, Zagreb, CroatiabSurgery Clinic, University Hospital „Sveti Duh“, Zagreb, Croatia

Background-aim

The aim of this study was to assess the diagnostic accuracy ofsome hematological laboratory parameters: platelets (Plt), meanplatelet volume (MPV), red cell distribution width (RDW), neutro-phil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio(PLR), in the diagnosis of acute appendicitis (AA) in the adultemergency department population.

Methods

The study was performed according to the 2015 Standards forReporting Diagnostic accuracy studies Guidelines. We consecutivelyincluded 78 patients admitted to the Emergency Department (ED)with suspected acute appendicitis. Parameters of the complete bloodcount were measured on Advia 2120i automated hematologyanalyzer (Siemens, Enlargen, Germany). The difference betweentwo groups was tested with Mann-Whitney test. Logistic regressionanalysis was performed to identify the significant predictors of AA,among the proposed laboratory parameters. Receiver operatingcharacteristic (ROC) curve analysis was used to determine theoptimal cut-off value of the identified significant predictors.Statistical analyses were performed using MedCalc Statistical Soft-ware version 16.2.0 (MedCalc Software bvba, Ostend, Belgium). P b

.05 was defined as the threshold of significance.

Results

In total 55/78 patients, in whom AA diagnosis was excluded, weredischarged, while 23/78 patients underwent a surgery for AA. In 18/23 patients who underwent the surgery, AA diagnosis was confirmedby histological analysis. There was no statistically significantdifference in Plt, MPV and RDW between groups. Although NLR andPLR were significantly higher in the confirmed AA group comparedto the discharged patients group (8.5 vs 2.65, and 25.55 vs 10.35, P b

.001, respectively), multivariate analysis identified only NRL as asignificant contributor to AA diagnosis (P b .001). NRL ROC analysisrevealed AUC = 0.874 (95% CI = 0.779–0.938) at a cut-off N4.4 (P b

.001) with a sensitivity and specificity of 89% (95% CI = 65.3–98.6)and 78% (95% CI = 65.8–87.9), respectively.

Conclusions

Plt, MPV, RDW and PLR were not useful parameters fordiagnosing acute appendicitis in the adult ED population. NLR abovethe proposed cut-off N4.4 can be useful aid in the differentiationbetween the patients with AA and symptomatic individuals withother diagnoses.

doi:10.1016/j.cca.2019.03.798

T411

CRP levels as predictor of admission to ICU in patients withpancreatitis acute

C. Rodríguez-Rojasa, P. Pérez-Cañadasa, V. Ramos-Arenasa, R.Cárdenas-Gámeza, A. García Rosc, S. Moran Sánchezb, M.D.Albaladejo-Otóna, L. García De Guadiana-RomualdoaaDepartment of Clinical Biochemistry, Hospital Universitario SantaLucia, Cartagena, SpainbDepartment of Gastroenterology and Hepatology, HospitalUniversitario Santa Lucia, Cartagena, SpaincDepartment of General and Communitarian Medicine, HospitalUniversitario Santa Lucia, Cartagena, Spain

Background-aim

Biomarkers can be useful tools for predicting the prognosis inpatients with acute pancreatitis (AP). Guidelines recommend a CRPlevel on admission ε 15 mg/dL as an analytical pancreatic alarm signindicating that patients may fail to recover satisfactorily. In this studywe have evaluated the value of CRP levels on admission toemergency department (ED) to predict Intensive Critical Unit (ICU)admission.

Methods

We performed a retrospective study including adult patientsadmitted to the ED with a diagnosis of AP. In all of them, CRP wasmeasured on presentation and at 48 h, by a nephelometric orturbidimetric immunoassay. Predictive value of CRP for outcomeswas evaluated by ROC AUC analysis and sensitivity, specificity andpredictive values for the above cutoff (15 mg/dL) were calculated.SPPS vs. 21 and Med Calc vs. 11 were used for statistical analysis.

Results

During the study period, 203 patients with a final diagnosis of APwere admitted to the ED. 8 patients were excluded because CRP wasnot measured in blood sample on admission. Finally, populationstudy included 195 patients (mean age (SD): 64 years (19; male: 99(50.8%). 35 (17.9) patients requiring admission to ICU. CRP levelswere significantly higher in patients requiring ICU management(mean CRP (SD): 8.1 mg/dL (11.1) vs. 2.6 (4.2); p b .001). AUC ROCfor CRP was 0.604 (95% Confidence interval (CI): 0.484–0.723; p b

.055). For a CRP level ε 15 mg/dL, sensitivity, specificity, predictivevalue positive and negative were 25.7 (95%CI: 9.8–41.6), specificity:96.3 (95%CI: 93.0–99.5), 60.0 (95% CI: 31.9–88.1) and 85.6 (95% CI:80.1–91.0).


Conclusions

Accuracy of CRP, measured on admission to ED, for predicting therequirement of ICU management, was low. Its performance do notsupport the use of this biomarker as criteria to rule-out or rule-in forthe defined outcome, with predictive and negative predictive valuebelow 95%.

doi:10.1016/j.cca.2019.03.799

T412

Hyperlipasemia without pancreatitis, is it possible?

R. Wijngaard, N. Rico, J.L. BediniCORE Laboratory, CDB, Hospital Clinic Barcelona, Spain

Background-aim

Hyperlipasemia (HL) is a common finding in pancreatic disorders.However, in our hospital we have observed high serum lipase withnormal amylase levels in patients without clinical signs andsymptoms of acute pancreatitis. These findings take clinicians to docomplementary tests which don't provide additional diagnostic valueand, in many cases, the results are perceived by mistake as analyticalerrors.

The aim of our study was to review the incidence of HL notrelated to pancreatic diseases in our hospital and identify the specificcause in each case through a literature research.

Methods

We reviewed, retrospectively, 264 patients from April 2017 toJune 2018 with lipase level more than three times the upper limit ofnormal (ULN) associated with amylases less than two times the ULN.Patients were excluded if they presented pancreatic diseases, biliarytract diseases, high amylase levels or in case of incomplete data.Patients were classified by the most probable cause of HL asdescribed in the literature. Other recorded data were: patient'sdemographics, blood test results, abdominal imaging findings andfollow-up laboratory test results.

Results

100 patients were included, mostly males (66%) with an averageage of 57.3 years. The average lipase level was up to 1680 +/−595.4 U/L (reference range b 393 U/L) and an amylase level 149 +/−40.4 U/L (reference range 20–104 U/L). In 71 patients there was afollow-up until normalization and in 63 patients an imaging test wasperformed, with no abnormal results. The most common diseasesidentified with lipase elevation were: renal failure (17%), neurosur-gical diseases (14%), gastrointestinal diseases (12%), drug-inducedHL (11%) and septic shock (7%). However, in most patients (18%) weobserved various factors that could cause the HL. In two patients noexplanation was found.

Conclusions

HL can be found in patients without pancreatic disorders. Themost prevalent diseases in our hospital were renal failure, neuro-surgical diseases and gastrointestinal diseases. Laboratory specialistsand clinicians need to be aware of these cases to make a more correctdifferential diagnosis and discard pancreatic disorders, especially

acute pancreatitis, if patients don't present other specific diagnosticcriteria.

doi:10.1016/j.cca.2019.03.800

T413

Validation of fatty liver index in a healthy Korean population andits comparison with triglyceride glucose index and its relatedparameters

J. LimDepartment of Laboratory Medicine, Chungnam National UniversityHospital, Daejeon, South Korea

Background-aim

NAFLD (nonalcoholic fatty liver disease) has become an emergingpublic health concern because of its high prevalence in both Westernand Asian countries. Many parameters were suggested for easydetection of NAFLD, but fatty liver index (FLI) is the most validated indifferent ethnic groups until now. Recently, however, triglycerideglucose (TyG) index and related parameters which combine obesityindices such as body mass index (BMI) and waist circumference(WC) were suggested as possible indices for NAFLD detection.Therefore, in this study, we sought to validate FLI in a healthyKorean population and to compare it with TyG index and TyG-related parameters, namely, TyG-BMI, and TyG-WC.

Methods

Data were collected from subjects who had undergone clinical,biochemical, and anthropometric assessment for annual health check-up at Chungnam National University Hospital, from 2014 to 2017. Atotal of 7162 subjects (4090 men and 3072 women) were included.Ultrasonography was used to diagnose fatty liver. Logistic regressionanalysis was performed after adjusting for confounding factors, tocompare and identify the associations of four parameters (FLI, TyGindex, TyG-BMI, and TyG-WC) with NAFLD. For each parameter, theodds ratios (ORs) and 95% confidence intervals (CIs) of quartiles 2–4were calculated and compared with those of quartile 1, as a reference.A receiver operating characteristic (ROC) curve analysis was con-ducted to evaluate the capability of each parameter to predict insulinresistance. Optimal cutoff points in the diagnosis of NAFLD weredetermined based on the maximum values of Youden's index.

Results

The adjusted ORs (95% CIs) for NAFLD were 64.3 (56.7–81.7) forFLI, 16.1 (13.4–19.2) for TyG index, 39.2 (31.6–48.6) for TyG-BMI,and 64.7 (50.9–82.1) for TyG-WC. The areas under the ROC curve foreach parameter were 0.843 (0.835–0.852) for FLI, 0.786 (0.777 to0.796) for TyG index, 0.837 (0.828 to 0.846) for TyG-BMI, and 0.841(0.832 to 0.849) for TyG-WC. Optimal cutoffs for NAFLD were 20 inFLI, 0.852 for TyG index, 201.46 for TyG-BMI, and 697.48 for TyG-WC.

Conclusions

FLI could be used to detect NAFLD accurately in Koreans but witha lower cutoff. In addition, TyG-WC was found to be comparable toFLI in NAFLD detection; thus, it can also be used for NAFLD detection.

doi:10.1016/j.cca.2019.03.801


T414

Role of the medicine laboratory in the development of newtherapeutics: A non-clinical study on the use of erythropoietin ina model of inflammatory bowel disease

R. Pintoa,b, V. Mateusa, A. Centenoc, C. Cardosoc, C. Domingosc, S.Pintoc, J. PecegueirocaiMED, ULisboa, Faculdade de Farmácia Universidade de LisboabJCS, Dr. Joaquim Chaves, Laboratório de Análises Clínicas, Miraflores –AlgéscJCS, Dr. Joaquim Chaves, Laboratório de Análises Clínicas, Miraflores –Algés

Background-aim

Erythropoietin in addition to being the main regulator oferythropoiesisis able to inhibit the activation of Nuclear FactorKappa B (NF-kB), due to his pleiotropic properties, therefore beingassociated with anti-inflammatory effects. Inflammatory boweldisease is a chronic disease whose quality of life is compromisedand where current therapy merely induces or keeps the patient inremission. For this reason we must continue to promote researchassociated with the treatment of this disease through the study ofnew pharmacological approaches.

Methods

The main goal of this study was to evaluate the effect oferythropoietin in a induced colitis model with 2,4,6-Trinitrobenzenesulfonic acid (TNBS) in mice with normal intestinalflora.

Mice with TNBS-induced colitis were treated daily with 500 UI/Kg/day (G500) or 1000 UI/Kg/day (G1000) of erythropoietin (closeto those used in clinical practice) administered intraperitoneally for4 days. The control group (GC) consisted of mice where only wasadministered TNBS (induction of colitis without erythropoietintreatment).

Results

Concerning the symptoms/signs, erythropoietin attenuated thereduction of body weight, reduced diarrhea and anus edemacompared to GCin a dose-dependent manner. The anti-inflamma-tory properties of erythropoietin in TNBS-induced colitis wereconfirmed by suppression (p b .05 vs GC) of pro-inflammatorymediators such as: Tumor Necrosis Factor alpha (TNF-alpha: pg/mL)(GC: 250 ± 40 vs G500: 110 ± 10 vs G1000: 90 ± 5),Interleu-kin-1 beta (IL-1beta: pg/mL))(GC: 260 ± 30 vs G500: 108 ± 9 vsG1000: 80 ± 7) and Myeloperoxidase (MPO: ng/mL))(GC: 42 ± 5vs G500: 22 ± 4 vs G1000: 12 ± 3) as well as through thesignificant increase (p b .05 vs GC) of the anti-inflammatorycytokine, Interleukin-10 (IL-10: pg/mL))(GC: 35 ± 3 vs G500: 65± 5 vs G1000: 145 ± 20). The treated mice also showed areduction (p b .05 vs GC) in the Alkaline phosphatase value(ALP:IU/L))(GC: 72 ± 2 vs G500: 45 ± 5 vs G1000: 39 ± 2), suggesting abeneficial effect of erythropoietin on TNBS-induced enterocyteinjury. The histopathological score (HS) is reduced (p b .05 vs GC)after treatment with erythropoietin, decreasing the severity andextent of colitis)(HS - GC: 17 ± 2 vs G500: 12 ± 2 vs G1000: 11 ±1.5).In addition, no alterations were observed in the renal andhepatic biomarkers (urea, creatinine, aspartate aminotransferase(AST) and Alanine aminotransferase (ALT)), as well as in thehematocrit.

Conclusions

In conclusion, erythropoietin reduced the inflammatory responseassociated with TNBS in a model of colitis in mice and may beexplored in the near future in clinical trials as a new pharmacologicaltherapy for the treatment of Inflammatory Bowel Disease.

doi:10.1016/j.cca.2019.03.802

T415

Cumulative levels of fecal hemoglobin for improving colorectalcancer screening management

J.M. Augea, H. Hurtadoa, A. Garciaa, L. Carotd, J. Graue, A. Buronb, M.Pellisec, C. Hernándezb, I. Torae, X. Castellsb, F. Maciab, A. Castellsc, X.BessadaBiochemistry and Molecular Genetics Department, HospitalClinic,Barcelona, SpainbEpidemiology and Evaluation Department, Hospital del Mar, Barcelona,SpaincGastroenterology Department, Hospital Clínic, Barcelona, SpaindGastroenterology Department, Hospital del Mar, Barcelona, SpainePreventive Medicine and Hospital Epidemiology Department, HospitalClínic, Barcelona, Spain

Background-aim

Fecal Immunochemical Test (FIT) for hemoglobin is used incolorectal cancer screening programmes to select those participantsundergoing to colonoscopy. However excretion of hemoglobin tofaeces (fHb) can be due to difference causes other than neoplasia.Even more, FIT pre analytics have sources of variability and theconcentration of fHb is related with age and gender. And also hasbeen demonstrated that an individual may present a positive resultfollowed by a negative result or vice versa only in a few days. Ourhypothesis is that the cumulative concentration of fHb in consecutivecolorectal cancer screening rounds might be useful to detect thoseindividuals with high risk neoplasia (HRN), even among FIT negativeparticipants.

Methods

We have evaluated 1771 average-risk participants (50–69 yearsold) from the Barcelona colorectal cancer screening programme witha negative FIT (b 20 μg Hb/g faeces) in the first and second round andwith a positive test in the third round. We have established threegroups based on the mean fHb concentration of the two first rounds(b 4 μg/g; 4–9 μg/g; N9 μg/g). This classification, in combination withgender and age, has been correlated to endoscopic findings. Theprincipal outcome has been HRN defined as colorectal cancer and/or5 ε adenoma / serrated lesions and/or lesion size ε20 mm.

Results

A multivariate logistic regression analysis identified men: oddsratio (OR) 1.68; 95% confidence interval (CI) (1.29–2.20), age (60–69 years old) OR 1.43; 95% CI (1.07–1.91) and mean fHb concentra-tion of the first two rounds (N10 μg/g) OR 3.47; 95% CI (2.07–5.81)as independent predictive factors for HRN. Combining these factors,different risk categories have been established. 12.96-fold (95% CI.4.17–40.27) higher risk of HRN was found between the twoextremes categories. The likelihood for HRN ranged from 6.3% to


46.7%. Interestingly, the likelihood for rule out CRC in the lowest riskcategories, ranged from 97.1% to 98.7%.

Conclusions

Cumulative fHb concentration in combination with gender andage in FIT negative participants could be useful to rule in or rule outHRN in subsequent screening rounds and help designing strategiesfor the management of the programmes in order to enhance theirefficiency.

doi:10.1016/j.cca.2019.03.803

T416

Diagnostic accuracy and usefulness of intraepithelial lymphocytesimmunophenotyping in intestinal mucosa for the diagnosis ofceliac disease

X. Gabaldó Barriosa,b, S. Cladellas Núñeza,b, M. Juanpere Aixalàa,b, I.Fort Gallifaa,b, L. Castro Reyesa,b, E. Martínez Sáncheza,b, F.J. MartínezCerezob, G. Castillejo-De Villasanteb, J.M. Simó I. Sisóa,baLaboratori de Referència Sud-Laboratori de Referència de CatalunyabUniversity Hospital Sant Joan de Reus

Background-aim

There is evidence that intraepithelial lymphocytes (IEL) play animportant role in the pathogenesis of celiac disease (CD). Flowcytometry is a powerful analytical tool for the study of smallintestinal immune cells and in particular the IEL. An IEL patterntypical of CD was defined, consisting of both an increase in ©™ + IELand a dramatic decrease in CD3- IEL. The aim of this study was todeterminate the normal range of IEL populations from a universityhospital and its diagnostic usefulness in active CD.

Methods

One hundred and twenty-five consecutive patients (mean age,18.9 years, range 1–63 years) who underwent small intestinal biopsyunder clinical suspicion of CD were prospectively included in theperiod July 2017–November 2018. The diagnosis of CD was based onESPGHAN and AGA criteria. IEL cells are isolated from intestinalbiopsies in Ca 2 +−free RPMI medium supplemented with 10%Fetal Calf Serum and calcium chelant EDTA at ~1 Mm for 60 min withcontinuous rotation at 12 rpm in a vertical shaker at roomtemperature. These cells in suspension are then washed and labeledfor flow-cytometric analysis. The antibodies used to define thedifferent IEL subsets were CD103-FITC, TCR©™-PE, CD45-APC andCD3-PercP(BD Biosciences). Statistical analysis was carried outwith the use of SPSS software (version 23, SPSS). A receiver operatorcharacteristic (ROC) curve analysis was carried out to determinethe optimal cut off (CO) values for the combination of ©™+ andCD3- IELs counts that could be used for the diagnosis of CD. Thediagnostic value of IEL lymphogram was calculated as sensitivity andspecificity.

Results

Among the 125 patients included in the study, 68 were children(b14 years, 50 celiacs) and 47 were adults (16 celiacs). From the ROCcurve analysis performed of IEL lymphogram in active CD diagnosis

showed that the optimal CO values for the percentages of ©™+ andCD3- were: 10.25% (sensitivity = 95% and specificity = 90%, AUC=0.944) and 8.13% (sensitivity = 87% and specificity = 88%, AUC=0.915), respectively.

Conclusions

The cut-off values of the IEL population analyzed has beenestablished in this study. Our results indicate that the evaluation ofIELs subsets could be useful to confirm diagnosis of celiac disease.

doi:10.1016/j.cca.2019.03.804

T417

Retrospective study of the results obtained in a colorectal cancerprevention program

V. O'valle Aísa, R. Jañez Carrera, L. De La Hoz Gil, N. HernándezMartínezBiochemistry Department, University Hospital Severo Ochoa, Leganés,Spain

Background-aim

Colorectal cancer (CRC) is the second leading cause of cancer-related deaths in our country and the most common malignanttumor considering both sexes. The nonivasive quantitative fecalimmunochemical test (FIT) for human hemoglobin is widelyrecommended and used as a biomarker in screening of CRC. CRCscreening program in our region is aimed at asymptomatic average-risk subjects, aged 50 to 69 years. Early detection and removal ofpreneoplastic lesions have consistently demonstrated efficacy inreducing the incidence and mortality.

Correlation between fecal hemoglobin concentration values andcolonoscopy. Assessment of the impact of age and gender to specificcut-off values of FIT in colorectal cancer screening.

Methods

We included all FIT-positive patients (cut-off 117 μg/L [23μghemoglobin/g faeces]), from May to October 2018. We reviewedcolonoscopies and pathologycal anathomy reports conducted for apositive FIT, classifying patients according to the diagnosis in 6categories: cancer colorectal (CRC), noninvasive high grade neoplasia(NHGN), nonivasive moderate grade neoplasia (NMGN), noninvasivelow grade neoplasia (NLGN), hyperplastic polyps (HP) and normal.

Fecal hemoglobin concentration was performed using SEN-TiFIT®270 analyzer (Sysmex).

Results

The study enrolled 6900 patients remitted from CRC preventionprogram in our area, which 430 (6,4%) were FIT positive; 280(65,1%) corresponding to men (mode age 65) and 150 (34,9%) towomen (mode age 67). Hemoglobin concentration (μg/l) medianfor men: CRC (2004,63), NHGN (2128,35), NMGN (1817,36), NLGN(534,72), HP (634,64); normal (500,91); for women: CRC(1239,36), NHGN (1890,99), NMGN (1074,67), NLGN (635,30), HP(844,33); normal (500,91). Using Kruskal–Wallis analyses wasobserved statiscally significantly different median FIT amongcategories (p b 0,0001).


Conclusions

We confirmed significantly higher values of fecal hemoglobincorrelates with the severity of histopathologic findings so theassessment of FIT facilitate grading of CRC-stages before thecolonoscopy. Cancer incidence increases with advancing age, thereare more positive results in patients over 61 years. It was found thatFIT concentration is higher in men in all the categories, thereforeage/sex-specific thresholds have potential value for screening in CRC.

doi:10.1016/j.cca.2019.03.805

T418

MALDI-TOF/MS peptidomic profiling for the diagnosis of inflam-matory bowel diseases

A. Padoanc, R. D'incàd, M.L. Scapellatob, G. Arrigonia, P. Cornoldic, M.Plebanic, D. BassocaDepartment of Biomedical Sciences, University of Padova, Padova, ItalybDepartment of Cardiologic, Thoracic and Vascular Sciences, PreventiveMedicine and Risk Assessment Unit, University Hospital of Padova,Padova, ItalycDepartment of Medicine, DIMED, University Hospital of Padova,Padova, ItalydDivision of Gastroenterology, University Hospital of Padova, Padova,Italy

Background-aim

Crohn's disease (CD) and ulcerative colitis (UC) represent the twomain forms of inflammatory bowel diseases (IBD). Fecal calprotectinis the most useful biomarker for IBD diagnosis currently available,despite it suffers limitations in both sensitivity (80%) and specificity(65%). This study was focused on the identification of new diagnosticMALDI-TOF/MS peptidomic profiles for improving IBD diagnosis,thus reducing the number of unnecessary colonoscopy.

Methods

Stool samples obtained from subjects without (C) or with IBDwere collected from the Department of Occupational Medicine andfrom the Department of Gastroenteroloy of the University-Hospitalof Padova, respectively. Samples were resuspended in water 1000:1(w/v), vortexed and ultracentrifuged for removing residual debris.Supernatants were mixed with acetonitrile 1:1 (v/v), allowing theprecipitation of abundant proteins. After ultracentrifugation, super-natants were evaporated, resuspended in 0.1% TFA and desalted.MALDI-TOF/MS analyses were performed in a m/z ranging from 1000to 4000 Da.

Results

Thirty-three and 133 stool samples from C and IBD, respectively,were collected. After MALDI-TOF/MS analyses, by evaluating all massspectra, a total of 438 features were identified. In C, 67 of the 79identified features were shared with IBD. In IBD, 359 features, insingle or in combination, were present in 111/133 patients. Theoverall analyses of all mass spectra allowed diagnose IBD with 83%sensitivity and 100% specificity. Moreover, 34 and 25 peptides wereclosely correlated with CD and UC respectively, allowing distinctionbetween the two diseases with 80% sensitivity and specificity. A

patent was deposited (Patent Number: 102018000005689, 24/05/2018, Ministero dello Sviluppo Economico - Ufficio Italiano Brevetti eMarchi).

Conclusions

The MALDI-TOF/MS peptidomic profiling not only represents aninexpensive, rapid, high throughput and sensitive method foranalyzing stool samples but it also allows to achieve bettercharacteristic than fecal calprotectin for the diagnosis of IBD andfor distinguishing CD from UC.

doi:10.1016/j.cca.2019.03.806

T419

Calprotectin and calgranulin C serum levels correlate withcomplications of pancreatic surgery

L. Pospíšilováa, E. Bartákovác,e, E. Koblihovád, M. Blahutováa, L.Kubátovád, M. Holubb, M. RyskadaDepartment of Clinical Biochemistry, Military University HospitalPrague, Czech RepublicbDepartment of Infectious Diseases, First Faculty of Medicine, CharlesUniversity and Military University Hospital Prague, Czech RepubliccDepartment of Infectious Diseases, First Faculty of Medicine, CharlesUniversity and Military University Hospital Prague, Czech RepublicdDepartment of Surgery, Second Faculty of Medicine, Charles Universityand Military University Hospital Prague, Czech RepubliceDepartment of Clinical Microbiology, Military University HospitalPrague, Czech Republic

Background-aim

Pancreatic surgery for cancer and chronic benign pancreaticprocesses are associated with significant morbidity and mortality.The sensitivity and specificity of currently used biomarkers indicat-ing postoperative complications is limited. The aim of the study wasto evaluate serum levels of calprotectin and calgranulin C in patientsundergoing pancreatic surgery and to test these proteins as potentialpredictors of postoperative complications.

Methods

Serum levels of calprotectin and calgranulin C were evaluated inadult patients (n= 98) undergoing pancreatic surgery: patients withthe cancer (n= 80), benign pancreatic processes (n = 13) andpreacancerous lesions (n= 5). The baseline serum levels of bothproteins were measured the day before the surgery. After thesurgery, measurements of calprotectin and calgranulin C in serumwere performed on days 1, 3, 5 and 7.

Results

The baseline serum levels of calprotectin and calgranulin C werefound to be significantly higher in patients when compared to healthycontrol (n = 26). After the surgery, the serum levels of both proteinswere significantly up-regulated over whole seven days of the studyperiod when compared to their baseline values. Interestingly,calprotectin and calgranulin C serum levels were significantly higherin patients with surgical complications (n= 37) in comparison topatients with uneventful recovery (n= 61) on days 3, 5 and 7.


Conclusions

Calgranulin and calprotectin C serum levels demonstratedsignificant diagnostic sensitivity and specificity in predicting devel-opment of surgical complications, which was slightly worse than thatof amylases in drain fluid but better than serum C-reactive proteinlevels and white blood cell count. These results suggest thatpancreatic cancer, chronic benign pancreatic processes, surgicalinjury of pancreatic tissue and postoperative complications can up-

regulate calprotectin and calgranulin C serum levels. Therefore, bothproteins should be extensively evaluated as potential biomarkers ofpathologic pancreatic processes.

The study was supported by Ministry of Health of the CzechRepublic, grant nr. 15-386A, and institutional support project SVV260369.

doi:10.1016/j.cca.2019.03.807


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