Clinical Chemistry Department Research Articles

The Incretin Response After Successful Islet Transplantation

The Incretin Response After Successful Islet Transplantation

Urinary Retinol Binding Protein Is a Marker of the Extent of Interstitial Kidney Fibrosis

Currently, a non-invasive method to estimate the degree of interstitial fibrosis (IF) in chronic kidney disease is not available in routine. The aim of our study was to evaluate the diagnostic performance of the measurement of urinary low molecular weight (LMW) protein concentrations as a method to determine the extent of IF. The urines specimen from 162 consecutive patients who underwent renal biopsy were used in the analysis. Numerical quantification software based on the colorimetric analysis of fibrous areas was used to assess the percentage IF. Total proteinuria, albuminuria, and the urinary levels of retinol binding protein (RBP), alpha1-microglobulin (α1MG), beta 2-microglobulin (β2MG), transferrin, and IgG immunoglobulins were measured. There was a significant correlation between the degree of IF and the RBP/creatinine (creat) ratio (R2: 0.11, p<0.0001). IF was associated to a lesser extent with urinary β2MG and α1MG; however, there was no association with total proteinuria or high molecular weight (HMW) proteinuria. The correlation between IF and RBP/creat remained significant after adjustment to the estimated glomerular filtration rate, age, body mass index, α1MG, and β2MG. The specificity of the test for diagnosing a fibrosis score of >25% of the parenchyma was 95% when using a threshold of 20 mg/g creat. In conclusion, RBP appears to be a quantitative and non-invasive marker for the independent prediction of the extent of kidney IF. Because methods for the measurement of urinary RBP are available in most clinical chemistry departments, RBP measurement is appealing for implementation in the routine care of patients with chronic kidney disease.

Ulf-Håkan Stenman

Finland lies huddled in the Baltic Sea—a sliver of forests and lakes caught between Norway, Sweden, and the looming stretch of land that is Russia. It is no wonder that the Finns have maintained a sizable and well-trained army at all times. Ulf-Hakan Stenman did his military service in 1960, at the age of 19. When he wasn't performing drills and maneuvers with his brigade, he liked to draw. “His big dream when he was young was to be a car designer. He was making sketches of different cars all the time,” said Borje Thorstrom, who served with him and would become one of his closest friends. Occasionally, they would have contests to see who could take a machine pistol to pieces and put it back together again. Stenman, known to his friends as Uffen, was by far the fastest—a testament to his physical agility but also to his habit of thinking from first principles. “His logical thinking is extraordinary,” said Thorstrom, an engineer. Those deductive powers were evident years later when Thorstrom's wife Eva fell and injured her knee. Stenman, who is professor emeritus in the department of clinical chemistry at the University of Helsinki, called a few days later. Thorstrom told him about Eva's accident and that the local hospital had given her a plaster cast and sent her home. “Uffen said, ‘May I speak to her?’ … On the telephone, he made a completely different diagnosis. He sent her to Helsinki to a special hospital to get operated on,” Thorstrom said. It turned out she had torn three tendons. “Her doctor was there seeing her and couldn't make the right diagnosis but Uffen could do it over the phone. It's amazing—he has such a special grip on everything,” Thorstrom said. Stenman—who gave up his dream of designing …

Ektacytometry and genetic testing in hereditary red cell membrane disorders

International Journal of Laboratory HematologyVolume 36, Issue 1 p. 107-107 Letter to the Editor Ektacytometry and genetic testing in hereditary red cell membrane disorders A. Huisman, A. Huisman [email protected] Department of Clinical Chemistry and Haematology, University Medical Center Utrecht, Utrecht, The NetherlandsSearch for more papers by this authorR. van Wijk, R. van Wijk Department of Clinical Chemistry and Haematology, University Medical Center Utrecht, Utrecht, The NetherlandsSearch for more papers by this author A. Huisman, A. Huisman [email protected] Department of Clinical Chemistry and Haematology, University Medical Center Utrecht, Utrecht, The NetherlandsSearch for more papers by this authorR. van Wijk, R. van Wijk Department of Clinical Chemistry and Haematology, University Medical Center Utrecht, Utrecht, The NetherlandsSearch for more papers by this author First published: 06 September 2013 https://doi.org/10.1111/ijlh.12140Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat No abstract is available for this article. Volume36, Issue1February 2014Pages 107-107 RelatedInformation

Placental STOX1 Y153H mutation is associated with severe early onset Preeclampsia and low birth weight

s / Placenta 34 (2013) A1–A99 A55 P1.128. PLACENTAL STOX1 Y153H MUTATION IS ASSOCIATED WITH SEVERE EARLY ONSET PREECLAMPSIA AND LOW BIRTH WEIGHT Caroline Dunk , Marie Van Djik , Laura Anderson , Cees Oudejans , Stephen J. Lye 1,3 1 Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, ON, Canada; Department of Clinical Chemistry, VU University Medical Center, Amsterdam, The Netherlands; Departments of Obstetrics and Gynecology and Physiology, The University of Toronto, Toronto, ON, Canada Objective: Controversy exists over the strength of the association of the gain-of-function mutation of the STOX1 transcription factor (Y153H allele) with Preeclampsia (PE). The use of maternal blood may not be appropriate for genotyping studies as this gene is paternally imprinted leading to the maternal allele specific expression of STOX1 in the placenta; thus we hypothesise that placenta genotype is more informative. We have previously shown that the HH STOX placental genotype induces an EVT specific reduction in invasion through the upregulation of a-T catenin supporting our contention of a role for STOX1 in the pathogenesis of PE. Methods: Placental DNAwas extracted from a cohort of Severe early onset PE with an AGA baby (n1⁄415), Severe early onset PE with IUGR (n1⁄417), Late Onset PE with an AGA baby (n1⁄411) and Term controls (n1⁄415). Sequencing of the STOX1 PCR product was performed by ACGT Inc. Toronto. Statistical analyses were performed using R software. Results: Assessment of the Y153H genotypes revealed that there was a 3.58 fold increased risk of severe early onset PE cases both with and without IUGR in comparison to term if the placenta was STOX1 HH. The placental samples from pregnancies with late onset of PE did not show this genotype correlation and resembled the term controls. When a preterm control group matched for gestational age with the severe early onset PE cases was also included in the analysis there was a highly significant association of the STOX1HH genotype with low birth weight, (p<0.002, corrected for ethnicity, fetal sex and PE). A further cohort of 250 PE and 250 control maternal and fetal samples are currently under analysis. Conclusions: These data support our hypothesis that defective EVT invasion associated with increased STOX1 transactivation is a precipitating factor in the development of PE. Funding: This work was funded by the CIHR MOP82811 http://dx.doi.org/10.1016/j.placenta.2013.06.164 P1.129. ASSESSMENT OF CARDIAC PARAMETERS IN LARGE FOR GESTATIONAL AGE INFANTS IN MOTHERS WITH W 2 The Ottawa Hospital, Ontario, Canada; 3 The University of Ottawa, Ontario, Canada Introduction: A major trend in modern education has been the use of online instructional modules. The most notable example is the success of the Khan Academy, an online educational website which has several million viewers. These video modules are often made available free on video or file exchange sites such as youtube. The openness of these forums allows for rapid knowledge translation through the wide dissemination and global exchange of academic tutorials. A drawback is that the lack of confidentiality or control precludes use of direct clinical material. Thus abstract or conceptual fields such as mathematics, which can be taught using simple graphics, drawing and voice, are most amenable to online micro-tutorials as shown by the Khan Academy experience. Aim: The ability to map two-dimensional histological sections of placenta to the three dimensional structure is challenging due to the complexity of the branching structures and the numerous reflecting surfaces. We speculate that the education of pathology and obstetric trainees in this area would benefit from online instructional modules modeled after the Khan Academy tutorials since it is both an abstract and a visualization problem.

ATP release in pancreatic acini and effects on the P2X7 receptor in pancreatic stellate cells

s / Pancreatolog S92 Department of Clinical Chemistry, University of Ulm, Ulm, Germany Department of General Surgery, Ostalb-klinikum Aalen, Aalen, Germany Department of Internal Medicine, University of Ulm, Ulm, Germany Introduction: Factors mediating the effects of pancreatic stellate cells (PSCs) on pancreatic cancer cells (PCCs) have not been clearly identified. Aims: To investigate the role of PSCs in PCCs migration and to identify underlying mechanisms. Materials & methods: Effects of conditioned PSCs supernatant (PSCSN) and exogenous adhesive molecules on the biology of Panc1 and UlaPaCa cells were investigated by modified Boyden chamber assay, adhesion assay and single cell tracking assay, respectively. Integrin expression and focal adhesion kinase (FAK) phosphorylation were assessed by Western blot. Anti-integrin a2/b1 antibodies and FAK inhibitor (PF-573228) were used to demonstrate the involvement of collagen I. Results: PSC-SN dose-dependently induced PCCs trans-migration, mainly by improving adhesion and motility. PSC-SN mediated adhesion was a prerequisite for the stimulation on PCCs migration. As pure chemokines, PSC-SN was not sufficient to stimulate the trans-migration or motility of PCCs. In contrast to poly-L-lysine or fibronectin, collagen I alone showed resembling effects to PSC-SN on PCCs, including polarized morphology, facilitated adhesion, accelerated motility and transmigration. Both PSC-SN and collagen I induced haptokinesis of Panc1 and haptotaxis of UlaPaCa. Anti-integrin a2/b1 antibodies attenuated PSC-SN/collagen I-induced PCCs trans-migration and adhesion. PSC-SN or collagen I constantly enhanced FAK phosphorylation (Tyr397) in PCCs. PF-573228 diminished PSC-SN/collagen I-induced PCCs haptotaxis/ haptokinesis. Conclusion: Collagen I is the major mediator for PSC-SN induced hapto-migration of PCCs. Through collagen I binding to integrin a2b1 on PCCs, FAK signaling pathway is initiated. S-9 Abstract id: 186. ATP release in pancreatic acini and effects on the P2X7 receptor in pancreatic stellate cells Kristian Agmund Haanes, Ivana Novak. Department of Biology, Copenhagen University, Denmark Introduction: ATP is an extracellular signal released from all cells, including pancreatic acini. Released ATP may stimulate the surrounding pancreatic stellate cells (PSC). Aims: Our aim was to characterize whether pancreatitis-associated stimuli induce ATP release from acini and investigate the importance of ATP in regulating proliferation of PSCs. Materials & methods: Mouse acini and AR42J cells were studied and ATP release was detected with luciferase kit. PSCs were isolated from WT and the Pfizer P2X7 KO mouse using a selective attachment method. Proliferation was monitored with BrdU incorporation. Results: The potential pathophysiological stimuli such as mechanical stress, hypotonicity and the bile acid chenodeoxycholate (CDC) all induced ATP release from acini. Alcohol, however, did not. CDC (1 mM) induced significant ATP release up to 20.4 7.4 nM/10^6 cells/ml. Basal ATP release supported PSC proliferation, as it was inhibited by apyrase, an ATP/ADP hydrolytic enzyme. The proliferation rate was lower in P2X7 KO PSCs compared to WT cells. Also the PSC number from P2X7 KO pancreas was 50% lower than from WT ones. Exogenous ATP further stimulated proliferation in a concentration-dependent and100 mM gave max stimulation. ATP in mM concentration was lethal to PSCs, and this effect was absent in P2X7 KO PSC. Basal/stimulated proliferation and cell death could all be inhibited with the P2X7 antagonists. Conclusion: Together, the study shows that ATP release and purinergic signalling should be considered as important factors in pathophysiological processes in pancreas. Importantly, ATP and P2X7 receptors are regulators of PSC proliferation and death, and could be potential targets in pathologies involving pancreatic fibrosis. y 13 (2013) S2–S98

Dr. de Rotte, et al reply

To the Editor: We thank Dr. Ranganathan for her interesting comments1 on our work2. In our recently published prediction model for methotrexate (MTX) non-response in juvenile idiopathic arthritis (JIA)3, ABCB1 rs1045642 was described, indicating the relative importance of this polymorphism to predict nonresponse to MTX in JIA. Also, we were able to reproduce this finding in a prospective cohort of 387 adult patients with rheumatoid arthritis (RA) receiving MTX: the ABCB1 rs1045642 polymorphism showed an association with improved clinical response (lower Disease Activity Score-28; ß = −0.16, p = 0.001). We agree that finding genetic predictors for MTX-induced toxicity and gastrointestinal (GI) adverse events is equally as important as response, because toxicity limits the considerations of a dose increase or continuation of MTX, and … Address correspondence to R. de Jonge, Department of Clinical Chemistry, Erasmus MC, 's-Gravendijkwal 230, 3015 CE, Rotterdam, The Netherlands. E-mail: r.dejonge{at}erasmusmc.nl

1 Screening for prostate cancer: Results of the Rotterdam section of the European Randomized study of Screening for Prostate Cancer (ERSPC)

a Erasmus University Medical Center, Department of Urology, Rotterdam, The Netherlands; Comprehensive Cancer Center, Rotterdam, The Netherlands; c Erasmus University Medical Center, Department of Pathology, Rotterdam, The Netherlands; d Erasmus University Medical Center, Department of Clinical Chemistry, Rotterdam, The Netherlands; e Erasmus University Medical Center, Department of Public Health, Rotterdam, The Netherlands E U RO P E AN URO LOG Y 6 4 ( 2 0 1 3 ) 5 3 0 – 5 3 9

A lipidomics approach to identify disorders in phospholipid metabolism; MEGDEL syndrome unraveled

Frederic M. Vaz (1), Saskia B. Wortmann (2,3), Antoine H.C. van Kampen (4), Eva Morava (2,3), Arjan P.M. de Brouwer (6), Ron A. Wevers (2,5), Willem Kulik (1) and Ronald J.A. Wanders (1) (1) Laboratory Genetic Metabolic Disease, Departments of Clinical Chemistry and Pediatrics, Academic Medical Center, Amsterdam, The Netherlands (2) Department of Pediatrics (3) Institute of Genetic and Metabolic Disease (IGMD), Radboud University Nijmegen Medical Centre (RUNMC), Nijmegen, The Netherlands. (4) Bioinformatics Laboratory, Clinical Epidemiology, Biostatistics and Bioinformatics (KEBB), Academic Medical Center, Amsterdam, the Netherlands (5) Laboratory of Genetic, Endocrine and Metabolic Diseases (LGEM), Department of Laboratory Medicine (6) Department of Human Genetics, Radboud University Nijmegen Medical Centre (RUNMC), Nijmegen, The Netherlands

Phenotyping a new mitochondrial fatty acid oxidation deficient mouse model

s ingediend voor het Amsterdam Kindersymposium 2013 33 Phenotyping a new mitochondrial fatty acid oxidation deficient mouse model E.F. Diekman (1,2), M.Weeghel (2), R.J. Wanders (2), G. Visser (1), S.M. Houten (2) (1) Department of Metabolic diseases, Wilhelmina Children’s Hospital, UMC Utrecht, the Netherlands (2) Laboratory Genetic Metabolic Diseases, Departments of Clinical Chemistry and Pediatrics, Emma Children’s Hospital, Academic Medical Center, University of Amsterdam, the Netherlands INTRODUCTION Very long chain acyl-CoA dehydrogenase defi ciency (VLCADD) is an inherited disorder of mitochondrial long-chain fatty acid (FAO) beta-oxidation. Patients present with hypoglycaemia, hepatomegaly, cardiomyopathy and myopathy. Although the phenotype of the LCAD KO mouse closely resembles the phenotype in humans, this mouse model does not have myopathy. As mice have two enzymes for degradation of long-chain fatty acids (LCAD and VLCAD), we hypothesised that a model in which one VLCAD allele is deleted in the LCAD KO mouse might have myopathy. LCAD/VLCAD double KO mice are not viable [Cox 2001]. Our objective was therefore to phenotype the LCAD-/-; VLCAD+/mouse model. METHODS We cross-bred LCAD+/-; VLCAD+/mice with LCAD-/-; VLCAD+/+ and characterized the resulting 4 genotypes using metabolic cages. RESULTS Enzymatic activity was signifi cantly decreased and C14:1/C2 acylcarnitine ratio was signifi cantly increased in the LCAD-/-; VLCAD+/mice opposed to all other genotypes. Heat production was decreased in LCAD-/-; VLCAD+/+ and LCAD-/-; VLCAD+/mice, but the RER was not signifi cantly changed. Locomotor activity was reduced in the LCAD-/-; VLCAD+/opposed to all other genotypes. Creatine kinase (CK) levels –a marker for rhabdomyolysiswere measured in plasma of mice of all genotypes after fasting. CK levels were within the normal range of 0-100 U/L. CONCLUSION Our preliminary data (n=4 per genotype) suggest that although LCAD-/-; VLCAD+/mice have lower FAO fl ux compared to LCAD-/-; VLCAD+/+ mice, they do not have myopathy.

Endoplasmic reticulum stress signaling in patients with X-linked adrenoleukodystrophy

s ingediend voor het Amsterdam Kindersymposium 2013 91 Endoplasmic reticulum stress signaling in patients with X-linked adrenoleukodystrophy Rob Ofman, Malu-Clair vd Beek, Inge Dijkstra, Ronald Wanders and Stephan Kemp Laboratory Genetic Metabolic Diseases, Department of Clinical Chemistry and Pediatrics, Emma Children’s Hospital, Academic Medical Center, University of Amsterdam INTRODUCTION X-linked adrenoleukodystrophy (X-ALD) is the most common peroxisomal disorder with a prevalence of approximately 1 in 17.000 live births. Mutations in the ABCD1 gene that encodes ALDP, an ATP-bindingcassette (ABC) transporter located in the peroxisomal membrane, results in impaired peroxisomal beta-oxidation and the subsequent accumulation of very-long-chain fatty acids (VLCFA; ≥ 22 or more carbon atoms) in plasma and tissues. Of all male patients, 35% will develop childhood cerebral adrenoleukodystrophy (CCALD), a rapidly progressive demyelinating disease of the central nervous system between 3 and 10 years of age. Most X-ALD patients develop adrenomyeloneuropathy (AMN) between the third and fourth decade of life. AMN is characterized by slow progressive stiff ness and weakness of the legs, impaired vibration sense in the lower limbs, spincter disturbance and impotence, commonly associated wth primary adrenocortical and/or testicular insuffi ciency. At present, more than 500 mutations in the ABCD1 gene have been identifi ed but no genotype-fenotype correlation is observed. We tested if VLCFA induce endoplasmic reticulum stress signaling in cultured cell lines from X-ALD patients. METHODS Fibroblasts derived from patients with X-ALD were cultured in the presence of VLCFA and the eff ect on cell viability and levels of ER stress markers was measured. RESULTS A signifi cant increase in the level of some ER stress markers was observed when fi broblasts from X-ALD patients were cultured in the presence of VLCFA but not in controls. CONCLUSION Our results indicate that increased levels of VLCFA can induce ER stress in patients with X-ALD.

P-026 Frequency of thrombophilia in females with stroke in association with oral contraceptive use

s / Thrombosis Research 131, Suppl. 1 (2013) S71–S103 S83 P-025 Adrenomedullin is associated with vonWillebrand factor activity in non-pregnant but not in pregnant women M. Poyhonen-Alho1, R. Lassila 2, L. Joutsi-Korhonen2, G. Nicholls 3, R. Kaaja 4 1Department of Obstetrics and Gynaecology, Helsinki University Central Hospital/Jorvi Hospital, Helsinki, Finland; 2Coagulation Unit, Departments of Hematology and Clinical Chemistry (HUSLAB Laboratory Services), Helsinki University Central Hospital, Helsinki, Finland; 3Christchurch Hospital, University of Otago, Christchurch, New Zealand; 4Turku University and Satakunta Central Hospital, Pori, Finland Introduction: Adrenomedullin has many cardiovascular beneficial effects, such as vasodilation, angiogenesis and tolerance of cells to oxidative stress and hypoxic injury. In addition, adrenomedullin has shown to augment the release and production of tissue factor pathway inhibitor in human aortic endothelial cells suggesting a role in anticoagulation on endothelial surface. Objective: We aimed to study the nocturnal variability of coagulation and plasma adrenomedullin in gestational diabetes, normal pregnancy and in non-pregnant women. Methods:We studied 41 women with gestational diabetes, 22 healthy pregnant and 14 non-pregnant controls. We assayed serial nocturnal changes of plasma adrenomedullin and coagulation variables, including factor VIII activity (FVIII:C), von Willebrand factor ristocetin cofactor activity (vWF:RCo), antigen (vWF:Ag) and collagen binding activity (vWF:CB) at 12 pm, 4 am and 7 am. Results: Adrenomedullin was higher in pregnant compared to non-pregnant women with no difference between the pregnant groups. In gestational diabetes, FVIII:C was lower than in normal pregnancy. AM correlated strongly with vWF:Ag (ρ=0.83, P=0.000), vWF:RCo (ρ=0.80, P=0.001) and vWF:CB (ρ=0.68, P=0.010) in non-pregnant controls. In contrast, these correlations disappeared in both pregnant groups. No correlation of adrenomedullin to FVIII:C existed within the study groups. Conclusions: These findings strengthen the view of adrenomedullin as a relevant factor in human coagulation. Distinct correlations of adrenomedullin with vWF and FVIII, usually working in tandem, could suggest an unravelled mechanism which is involved in the coupling of vWF with FVIII. This finding needs more insight from future studies. P-026 Frequency of thrombophilia in females with stroke in association with oral contraceptive use P. Dulicek1, M. Kost’a 1, P. Sadilek1, M. Beranek2 1IVth Department of Internal – Hematology; 2Department of Clinical Biochemistry, University Hospital and Medical Faculty Hradec Kralove, Czech

Fluconazole-induced fixed drug eruption imitating herpes labialis with erythema multiforme

ejd.2012.1806 Auteur(s) : Zandra Nymand Jensen1 zjensen@health.sdu.dk, Anette Bygum2, Per Damkier1 1 Department of Clinical Chemistry and Pharmacology, 2 Department of Dermatology, Odense University Hospital, Winslowparken 19, 2, 5000 Odense C, Denmark A 38-year old woman was referred with suspected labial herpes accompanied by peculiar dusky red skin lesions on her legs. For 10 months, she had experienced 8-10 attacks of prickly nummular oedematous red skin lesions that had developed into persistent hyperpigmented [...]

OXA-23-producing Acinetobacter species from horses: a public health hazard?

Department of Pathology, Bacteriology and Avian Diseases, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium; Department of Bacteriology and Immunology, CODA-CERVA-VAR, Groeselenberg 99, 1180 Brussels, Belgium; Department of Surgery and Anaesthesiology of Domestic Animals, Faculty of Veterinary Medicine, Ghent University, Salisburylaan 133, 9820 Merelbeke, Belgium; Laboratory of Bacterial Genetics, National Institute of Public Health, Srobarova 48, 100 42 Prague, Czech Republic; Department of Clinical Chemistry, Microbiology and Immunology, Faculty of Medicine & Health Sciences, Ghent University, De Pintenlaan 185, Ghent, Belgium

Hypothermic cardiopulmonary bypass surgery in a 7-year-old boy with a cold agglutinin

Dear Editor, Cold agglutinins (CA) are predominately IgM auto-antibodies that optimally react with surface antigens on red blood cells (RBCs) at low temperatures. Subsequently, agglutination of RBCs can occur followed by complement fixation and haemolysis. CAs seldom cause significant intraand extravascular haemolysis under normothermic conditions. However, they could be of relevance in patients undergoing cardiac surgery when hypothermic cardiopulmonary bypass (CPB) and cardioplegia are used [1]. There are conflicting opinions in the literature about screening patients for CAs prior to hypothermic CPB. CAs are rare and low levels can be found in healthy individuals. The incidence of CAs in screened cardiac surgery patients is approximately 0.8–4 % [1–4]. Although serious complications (haemolysis, micro-vascular occlusion and organ failure) have been described, it has also been demonstrated that patients with low titre, low thermal amplitude CAs can safely undergo CPB [1–4]. Therefore, and following Dutch guidelines recommendations (CBO consensus Bloedtransfusie, http://www.cbo.nl/thema/Richtlijnen/ Overzicht-richtlijnen/Bloedtransfusie/), we do not screen for CAs in adults. However, as described also by others recently, we do perform screening for CAs in children undergoing cardiac surgery for which deep hypothermia is indicated [5]. A 7-year-old boy was referred for serological testing prior to hypothermic cardiac surgery. The patient’s blood group was O positive and antibody screening tests were negative. The cold agglutinin assay (saline, 16 °C) revealed the presence of a cold autoantibody with IH specificity. CAs are often directed against the Ii blood group collection [1]. Anti-IH antibodies have been implicated as the cause of a haemolytic transfusion reaction in a couple of cases [6–8]. CAs are generally produced in response to infections or secondary to a lymphoproliferative disorder [9]. Our patient had recently been treated with antibiotics for an infection. The importance of a CA is determined by the thermal amplitude (highest temperature at which the antibody binds to RBCs) and titre. The IH-antibody in our patient displayed wide thermal amplitude (28 °C; titre 1:2). Because of the associated risk of intravascular haemolysis during CPB, and the fact that postponing the elective surgery was safe for the patient, the procedure was cancelled. The presence of CAs is often transient and therefore the thermal amplitude of the antibody was determined periodically. The thermal amplitude decreased slowly to 24 °C after 3 months (titre 1:1) and 22 °C after 5 months (Fig. 1). After 7 months, the thermal amplitude of the antibody remained stable and we decided to perform the procedure under less extensive hypothermic conditions. The patient was cooled to 24.9 °C and surgery was performed successfully. The antibody did not show any reactivity at this temperature and accordingly, the patient did not show signs of hemolysis. The patient received 2 units of RBC, as a consequence of the CPB procedure, and had stable post-surgery haemoglobin of 11.3 g/dL, without any signs of hemolysis. In conclusion, CAs can cause haemolysis in patients undergoing hypothermic cardiac surgery. Although screening for CAs in all patients is not recommended it can be useful in specific patient groups (e.g. children undergoing I. Hubeek :W. W. van Solinge :K. M. K. de Vooght Department of Clinical Chemistry and Haematology, University Medical Center Utrecht, Utrecht, The Netherlands

Venipuncture Versus Peripheral Catheter: Do Infusions Alter Laboratory Results?

Our aim was to evaluate the equivalence between analytic parameters from blood samples obtained from a saline solution lock device used for the infusion of drugs and those from venipuncture. In our emergency department, patients bearing a saline solution lock device have blood extracted by venipuncture to avoid possible contamination of the sample. Adults from the emergency department with a saline solution lock device who required laboratory tests were selected as candidates for this cross-sectional observational study. Infusions were halted and flushed with 0.9% saline solution; 2 minutes later, 2 mL of blood was drawn and discarded, and the corresponding laboratory tubes were filled. Immediately after, another sample was withdrawn from the opposite extremity by venipuncture. Both samples were analyzed for hematology, biochemistry, venous blood gases, and coagulation parameters. Concordance was evaluated by use of the intraclass correlation coefficient with its 95% confidence intervals; Bland-Altman plots were used to illustrate the percentage of samples with differences exceeding 2 SDs. The mean differences were also checked to detect those exceeding the laboratory's systematic error. An intraclass correlation coefficient of over 0.9 was achieved for all parameters except for pH, partial pressure of carbon dioxide, and partial pressure of oxygen. Differences of over 2 SDs were found in fewer than 10% of all parameters. None of them exceeded 3 SDs, except for pH and venous blood gases. All parameters showed differences below the laboratory's accepted systematic error except for pH and venous blood gases. Blood samples extracted from a peripheral catheter with or without drug infusions are valid for the analysis of hematology, biochemistry, and coagulation parameters but not for venous blood gases. Nurses should know the benefits of using an existing peripheral catheter for drawing blood samples for laboratory analysis even when infusing commonly used drugs. Emergency nurses should consider collecting blood specimens from a venous access device regardless of the type of drug infusions administered, because it is a safe, simple, and fast technique, which is time efficient when treating patients with limited venous access sites. This procedure reduces patient discomfort and the risk of complications related to venipunctures.

Flow chamber-based assays to measure thrombus formation in vitro: requirements for standardization.

M. ROEST ,* A . RE IN INGER , J . J . Z W A GI NG A , M. R . K ING ,§ J . W. M. H EEMSKERK– and THE B IORHEOLOGY SUBCOMMITTEE OF THE SSC OF THE ISTH *Department of Clinical Chemistry and Haematology, University Medical Center Utrecht, Utrecht, the Netherlands; Department of Transfusion Medicine and Hemostaseology, University Clinic Munich, Munich, Germany; Department of Immunohematology and Bloodtransfusion, Leiden University Medical Center, Leiden, the Netherlands; §Department of Biomedical Engineering, Cornell University, Ithaca, NY, USA; and –Department of Biochemistry, Maastricht University, Cardiovascular Research Institute Maastricht, Maastricht, the Netherlands

Early detection of severe sepsis in the emergency room: Diagnostic value of plasma C-reactive protein, procalcitonin, and interleukin-6

To determine the diagnostic values of plasma C-reactive protein (CRP), procalcitonin (PCT), and interleukin-6 (IL-6) using an electrochemiluminescence immunoassay (ECLIA) method (Roche Diagnostics GmbH, Mannheim, Germany) to identify severe sepsis in an emergency room (ER) setting. This was a single-centre prospective follow-up study of 539 consecutive adult patients admitted to the ER with suspected infection. Blood samples were taken concurrently with blood cultures at admission. Patients were divided into 5 groups on the basis of systemic inflammatory response syndrome (SIRS) criteria, documentation of bacterial infection, and organ dysfunction. Fifty-nine patients with no SIRS or bacterial infection, 68 patients with bacterial infection but no SIRS, 54 patients with SIRS but no bacterial infection, 309 patients with sepsis (SIRS and bacterial infection), and 49 patients with severe sepsis (sepsis and organ failure) were evaluated. In a logistic regression model, the odds ratio (OR) for PCT was 1.58 (95% confidence interval (CI) 1.37-1.82, p < 0.0001), for IL-6 was 1.54 (95% CI 1.32-1.80, p < 0.0001), and for CRP was 1.33 (95% CI 1.01-1.75, p = 0.045). The area under the curve (AUC) was 0.77 (95% CI 0.71-0.84) for PCT, 0.72 (95% CI 0.64-0.80) for IL-6, and 0.60 (95% CI 0.51-0.69) for CRP. PCT emerged as the best marker for severe sepsis, but the difference in AUC was not significant between PCT and IL-6. In multivariate logistic regression analysis, after adjusting for confounders, PCT and IL-6 remained significant independent predictors of severe sepsis. PCT and IL-6 proved superior to CRP in detecting patients with severe sepsis. The findings thus support the use of either PCT or IL-6 as an early tool to diagnose severe sepsis. The automatic ECLIA method allows even night-shift measurements.

A life‐threatening episode of treatment‐resistant haemolysis in a pregnant patient with dyserythropoietic anaemia (CDA) type I

Transfusion MedicineVolume 22, Issue 2 p. 145-147 A life-threatening episode of treatment-resistant haemolysis in a pregnant patient with dyserythropoietic anaemia (CDA) type I I. Bank, Corresponding Author I. Bank Department of Immunohematology and Blood Transfusion, Leiden University Medical Center Sanquin Blood Supply Foundation, Division of Transfusion Medicine, Leiden, The NetherlandsI. Bank, MD, PhD, Sanquin Blood Supply Foundation, Division of Transfusion Medicine, Plesmanlaan 1A, 2333 BZ, Leiden, The Netherlands.Tel.: +31 71 5685075; fax: +31 71 5685191;e-mail: [email protected]Search for more papers by this authorA. A. M. Ermens, A. A. M. Ermens Department of Clinical Chemistry and Haematology, Amphia Hospital, Breda, The NetherlandsSearch for more papers by this authorJ. M. Van Der Linden, J. M. Van Der Linden Department of Clinical Chemistry and Haematology, Amphia Hospital, Breda, The NetherlandsSearch for more papers by this authorA. Brand, A. Brand Department of Immunohematology and Blood Transfusion, Leiden University Medical Center Sanquin Blood Supply Foundation, Division of Transfusion Medicine, Leiden, The NetherlandsSearch for more papers by this author I. Bank, Corresponding Author I. Bank Department of Immunohematology and Blood Transfusion, Leiden University Medical Center Sanquin Blood Supply Foundation, Division of Transfusion Medicine, Leiden, The NetherlandsI. Bank, MD, PhD, Sanquin Blood Supply Foundation, Division of Transfusion Medicine, Plesmanlaan 1A, 2333 BZ, Leiden, The Netherlands.Tel.: +31 71 5685075; fax: +31 71 5685191;e-mail: [email protected]Search for more papers by this authorA. A. M. Ermens, A. A. M. Ermens Department of Clinical Chemistry and Haematology, Amphia Hospital, Breda, The NetherlandsSearch for more papers by this authorJ. M. Van Der Linden, J. M. Van Der Linden Department of Clinical Chemistry and Haematology, Amphia Hospital, Breda, The NetherlandsSearch for more papers by this authorA. Brand, A. Brand Department of Immunohematology and Blood Transfusion, Leiden University Medical Center Sanquin Blood Supply Foundation, Division of Transfusion Medicine, Leiden, The NetherlandsSearch for more papers by this author First published: 16 August 2011 https://doi.org/10.1111/j.1365-3148.2011.01096.xCitations: 6Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat Citing Literature Volume22, Issue2April 2012Pages 145-147 RelatedInformation

Von Willebrand factor predicts major bleeding and mortality during oral anticoagulant treatment

Oral anticoagulation (OAC), predominantly with warfarin, is an effective treatment to prevent thromboembolic events. Serious bleeding is a frequent and feared treatment complication. In this longitudinal cohort study of OAC-treated patients, we aimed to evaluate the relationship between von Willebrand factor (VWF) levels and risk of bleeding complications, cardiovascular mortality and all-cause mortality. A total of 719 patients receiving warfarin treatment were observed for a mean duration of 4.2 years. All bleeding complications causing hospitalization were registered and classified into clinically relevant bleeding (CRB) and major bleeding. Ischaemic stroke, peripheral arterial embolism, myocardial infarction, and death were also recorded. We identified 113 cases of CRB and 73 of major bleeding. In total, 161 deaths occurred during follow-up with cardiovascular disease identified as the cause of death in 110 patients. Patients in the highest tertile of VWF had a significantly increased risk of bleeding complications: hazard ratio (HR) 2.53 (95% CI 1.41-4.56) for major bleeding and HR 2.19 (95% CI 1.38-3.48) for CRB. VWF, expressed either in tertiles or as a continuous variable, showed a significant association with cardiovascular mortality (HR 1.68, 95% CI 1.40-2.01) and all-cause mortality (HR 1.77, 95% CI 1.52-2.05). In multivariate Cox regression analysis, the findings remained significant after adjusting for age, high-sensitivity C-reactive protein and creatinine. Patients with high levels of VWF had an increased risk of bleeding complications, cardiovascular mortality and all-cause mortality during OAC treatment. Our findings imply that the use of VWF as a risk marker for thromboembolic events is complicated by the association of VWF with bleeding complications.