Donor-derived tuberculosis via orthotopic liver transplantation.

We present a case of donor-derived tuberculosis after liver transplantation, in which the donor origin of the Mycobacterium tuberculosis isolate was made most likely by DNA fingerprinting. Screening for latent tuberculosis of transplant donors originating from high endemic areas with an ex-vivo interferon-gamma release assay should be considered.

Organ Donation After Euthanasia in the Netherlands: A Case Report.

In 2014, there was still a shortage of available organs for transplantation, and 1044 patients were waiting for an organ in the Netherlands. Maximizing the pool of organ donors is part of the solution. In 2001, the Dutch Termination of Life on Request and Assisted Suicide Act was adopted, legalizing euthanasia under strict conditions. In 2010, 3136 reports were made of euthanasia and assisted suicide; in 2014, 5306 reports were made. Among them were patients with a desire to donate their organs after their deaths. Although a potential source of donor organs, only a few cases of organ donation after active euthanasia have been described. Since 2012, 16 combinations of these procedures have been performed in the Netherlands. The literature mentions 16 Belgian cases between 2005 and 2013. This limited number can be the result of lack of knowledge about this subject among healthcare professionals or because of practical, ethical, and/or legal considerations. Performing this combination has possible advantages, both in number as well as in transplantation outcomes. By describing a recent case in our center, we will try to outline the state of the art in the Netherlands and disseminate knowledge about the possibilities and limitations of organ donation after active euthanasia.

Quality of care and analysis of surgical complications.

BACKGROUND: During the past years evaluation of quality of care has become an important aspect of transparency of care, and complications is one of these parameters. Therefore, we analyzed the complication rate in an academic hospital over a 6-year period. METHODS: During the period 2004-2009, all adult surgical patients admitted to and discharged from the Department of Surgery were selected for this time trend study. The Dutch national surgical complication registry was used in the analysis, which registers according to a three-tiered matrix-like classification system. Yearly changes in complication rates were analyzed statistically using the χ(2) for trend test. Subsequently, multivariable regression analysis was used to find significant independent predictors for sustaining a complication. RESULTS: The mean complication rate per admission rose significantly from 0.18 in 2004 to 0.30 in 2009 (p < 0.001). The largest increase was observed by the following variables: less severe complications, complex surgical procedures, and ASA classification. Delirium, gastoparesis, and ileus were complications showing the largest increase. Age, male gender, ASA, and surgical complexity were found as independent predictors. CONCLUSIONS: This study showed a significant increase of complications. The increase was mainly due to less severe complications, in particular delirium, ileus, and gastroparesis.

Addressing patients' information needs: a first evaluation of a question prompt sheet in the pretreatment consultation for patients with esophageal cancer.

This study aims to develop and pilot a question prompt sheet to assist esophageal cancer patients to obtain desired information in the consultation in which potentially curative esophagectomy is discussed. Whether a prompt sheet affected patients' question asking, the number and scope of topics discussed, the length of the consultation, and patients' satisfaction is investigated. Patients (n= 30) were randomized either to receive care as usual (control group) or to receive a prompt sheet (intervention group). All patients completed a baseline questionnaire, their consultations were audio-recorded and content-coded, and they received a structured telephone interview 2 days after the consultation to assess satisfaction. Patients provided with the prompt sheet marked a median of 19 questions. They asked significantly more questions as compared with patients in the control group (median of 12 vs. 8 questions). Questions mainly addressed treatment options and procedures. No differences were found with regard to consultation length and patient satisfaction. Our results suggest that providing patients with a simple, easy-to-implement tool such as a question prompt is appreciated and helps patients to ask more questions during the consultation without increasing the length of the consultation.

Intrathoracic manifestations of cervical anastomotic leaks after transhiatal and transthoracic oesophagectomy.

BACKGROUND: A possible advantage of cervical oesophagogastrostomy over intrathoracic anastomosis after oesophagectomy is the presumed mild clinical course of cervical anastomotic leakage. The incidence and consequences of intrathoracic manifestations after cervical anastomotic leakage remain unclear, and were investigated in this study. METHODS: Consecutive patients undergoing potentially curative transhiatal oesophagectomy (THO) or transthoracic oesophagectomy (TTO) with cervical oesophagogastrostomy between 1993 and 2007 were included. Intrathoracic manifestations after cervical anastomotic leakage were compared following THO and TTO. Multivariable logistic regression analysis was used to identify potential risk factors for intrathoracic manifestations. RESULTS: Seventy-nine (15.8 per cent) of 501 patients developed anastomotic leakage after THO compared with 50 (15.3 per cent) of 327 after TTO (P = 0.853). Intrathoracic manifestations developed in 21 (27 per cent) and 22 (44 per cent) patients respectively (P = 0.041). A transthoracic approach was the only independent predictor of the development of intrathoracic manifestations in patients with cervical leakage (odds ratio 2.60; P = 0.022). Total hospital stay (P < 0.001), intensive care unit stay (P < 0.001) and in-hospital mortality (P = 0.035) were greater in patients with intrathoracic manifestations than in those without. CONCLUSION: Intrathoracic manifestations of cervical anastomotic leakage are associated with a prolonged hospital stay, carry a higher mortality and occur more frequently after TTO than THO.

The -589C>T polymorphism in the interleukin-4 gene (IL-4) is associated with a reduced risk of myocardial infarction in young individuals.

BACKGROUND: Inflammatory reactions contribute to the development of arterial disease. We investigated the role of interleukin-4 (IL-4) in the development of myocardial infarction (MI) by genotyping patients with MI and control subjects for the -589C>T (rs2243250) single nucleotide polymorphism (SNP), which tags a functional haplotype of IL-4. METHODS AND RESULTS: Study of Myocardial Infarctions Leiden (SMILE) included 560 men with a first MI and 646 control subjects. The Valencia study included 305 patients with MI at T genotype was found [odds ratio (OR) 0.84; 95% CI 0.37-1.95 for -589TT and 0.82; 95% CI 0.62-1.07 for -589CT compared with -589CC]. In patients younger than 50 years, carriership of one or two -589T alleles was associated with a reduced risk of MI (OR 0.57: 95% CI 0.34-0.95). This result was replicated in the Valencia study, where carriers of one or two -589T alleles had a reduced risk of MI (OR 0.67: 95% CI 0.47-0.95), with a strong protective effect of the -598T allele in homozygous -589T (OR 0.33: 95% CI 0.10-1.05). In the control subjects of the Valencia study, the -589T allele was associated with reduced levels of F1+2. CONCLUSION: Our data indicate that the IL-4 haplotype tagged by the -589T allele reduces the risk of MI in young individuals.

A multi-channel simultaneous data acquisition and waveform generator system designed for medical applications.

To meet the needs of our research programmes on auditory prostheses for the totally deaf, a 15-channel data acquisition and waveform generator system with flexible triggering, pacing and linking was developed. It allows both synchronous and asynchronous operation at high speed. Each channel of the system, which is controlled by a simple personal computer, has an on-board microcontroller, a 512 kWord signal memory, a voltage input and both voltage and current outputs. The system includes a master pacer and trigger unit with elaborate hardware and software triggering options. This paper describes the system hardware and the software used to control it. Finally, some of its application are demonstrated. The flexibility of the system makes it widely applicable in the field of biomedical engineering.

Heerlen polymorphism of protein S, an immunologic polymorphism due to dimorphism of residue 460.

We recently developed an enzyme-linked immunosorbent assay (ELISA) for total protein S (PS) antigen using the monoclonal antibody S-12. During the screening of thrombophilic patients we identified a patient, who was using marcoumar, with 0% PS by monoclonal ELISA and 23% PS by polyclonal ELISA. Further analysis of this patient and his family showed that the patient was a compound heterozygote for type 1 PS deficiency and for an abnormal PS molecule (PS-Heerlen) that was not recognized by the S-12 antibody. Similar observations were made in two sisters from an unrelated Dutch family. Subsequent studies showed that PS Heerlen has a slightly lower molecular weight (71,000) than normal PS (73,000), binds normally to C4b-binding protein, and retains full activated protein C cofactor activity. The alteration in the PS Heerlen molecule was identified as a substitution of Ser460 by Pro, which is due to a unique T---C transition in exon 13 of the active PS-alpha gene. The substitution occurs in the consensus sequence for the potential N-linked glycosylation of Asn458. Digestion with N-glycanase showed that normal PS probably contains three N-linked oligosaccharide side chains, while PS Heerlen contains only two (Asn458 not glycosylated?). Segregation analysis in the two original families showed that the presence of the genetic abnormality was always associated with the PS-Heerlen phenotype. The frequency of the PS-Heerlen allele was found to be 0.52% in the general population and 0.67% in a population of patients with unexplained thrombophilia. There is no evidence that the PS Heerlen allele is associated with an increased risk for thrombosis.

Two ELISA's for measurement of protein S, and their use in the laboratory diagnosis of protein S deficiency.

Protein S is a vitamin K-dependent coagulation factor involved in the regulation of the anticoagulant activity of activated protein C (APC). Heterozygosity for hereditary protein S deficiency is considered as a risk factor for the development of thrombotic disease. To measure the plasma concentration of protein S, two enzyme-linked immunosorbent assays (ELISA) have been developed: the ELISA-p (using polyclonal antibodies both as catching and tagging antibodies) and the ELISA-m (using polyclonal antibodies as catching antibodies and a monoclonal antibody as tagging antibody). The performance of these assays has been compared with that of an immunoradiometric assay (IRMA) for total protein S. Results obtained with both ELISA's in plasma's of healthy volunteers, patients with hereditary protein S deficiency type I and patients using oral anticoagulant treatment correlate very well with those of the IRMA: r = 0.974 (n = 57) for ELISA-p and r = 0.966 (n = 57) for ELISA-m. The intra- and inter-assay variation (n = 6) were calculated to be 5% and 11% for the ELISA-p and 4% and 10% for the ELISA-m. Both ELISA's can be used for the measurement of total protein S, but also for the measurement of free protein S and C4b-BP-PS complexes, after PEG precipitation. The ELISA-m was further used to measure free protein S in healthy volunteers (n = 40), patients with a protein S deficiency type I (n = 20) and patients with unexplained familial thrombosis (n = 76). In the first two groups measured and calculated free protein S levels were very similar. In the third group 7 patients were identified with total protein S in the normal range, but with measured free protein S and the ratio between the measured free protein S and calculated free protein S below the lower limits of the normal range.

A second plasma inhibitor of activated protein C: alpha 1-antitrypsin.

Inactivation of activated protein C (APC) in normal human plasma was studied in the absence and presence of heparin. In the absence of heparin APC inactivation followed pseudo-first order kinetics. In the presence of heparin the neutralization of APC was found to be biphasic. Up to 500 nM APC could be readily inactivated in normal plasma, indicating that the concentration of the APC inhibitor must be higher than previously assumed. Plasma deficient in the protein C inhibitor (PCI-I, as described by Suzuki and coworkers) and deficient in beta 2-glycoprotein I still possessed APC neutralizing capacity, presumably through the formation of complexes of APC with another plasma protein as was demonstrated by immunoblotting with anti-protein C antibodies. Together these data made us to conclude that a second inhibitor of APC (PCI-II) must be present in normal human plasma. This second inhibitor should be heparin independent, have a relatively high plasma concentration and form complexes with APC. Subsequently, we purified this PCI-II by isolating APC-PCI-II complexes from plasma deficient of vitamin K dependent proteins, PCI-I and beta 2-glycoprotein-I, to which purified human APC had been added. Purified PCI-II has a molecular weight of 50,000 daltons and aminoacid analysis revealed that PCI-II is identical with alpha 1-antitrypsin (alpha 1-AT). The second order rate constant for the reaction between purified alpha 1-AT and APC was found to be 269 M-1 min-1 in the absence of calcium and 602 M-1 min-1 in the presence of calcium.(ABSTRACT TRUNCATED AT 250 WORDS)

Immunoradiometric assays for the Ca(II)-dependent and Ca(II)-independent conformation of human protein C.

From a rabbit antiserum against human protein C, two subpopulations of antibodies recognizing Ca(II)-dependent and NonCa(II)-dependent epitopes of human protein C respectively were affinity purified for use in solid-phase immunoradiometric assays. The two assays were specific for PC:Ca(II)Ag and PC:NonCa(II)Ag and highly sensitive with a lower limit of detection of 2 ng/ml. The concentration of PC:Ca(II)Ag and PC:NonCa(II)Ag and their ratio in healthy volunteers was 0.91 +/- 0.16 U/ml, 0.95 +/- 0.14 U/ml and 0.97 +/- 0.18. In a group of patients treated with oral anticoagulants the ratio PC:Ca(II)Ag/PC:-NonCa(II)Ag was significantly decreased (to about 0.6) as compared with normal controls suggesting the presence of sub- and noncarboxylated protein C molecules (PIVKA-PC) in plasma of patients on oral anticoagulants. Analysis of plasmas of patients with a hereditary (n = 8) or isolated (n = 4) type II protein C deficiency and a history of thrombo-embolic disease did not reveal patients with a PC:Ca(II)Ag level and a PC:Ca(II)Ag/PC:NonCa(II)Ag ratio below the lower limit of the normal range suggesting that the frequency of genetic protein C variants with defects in the Ca(II)-dependent conformation is low.

Immunoradiometric assay for the calcium-stabilized conformation of human protein S.

Rabbit polyclonal anti-protein S serum was fractionated with immobilized human protein S to establish solid-phase immunoradiometric assays recognizing Ca(II)-dependent and NonCa(II)-dependent epitopes of human protein S. The two assays were specific for PS:Ca(II)Ag and PS:NonCa(II)Ag and highly sensitive with a lower limit of detection of about 2.5 ng/ml. PS:Ca(II)Ag and PS:NonCa(II)Ag levels were measured in immunopurified protein S, thrombin-modified protein S and chymotrypsin-cleaved protein S. Only in chymotrypsin-cleaved protein S an important discrepancy between the two antigen levels was observed. Ranges for the concentration of PS:Ca(II)Ag and PS:Non-Ca(II)Ag and their ratio were established in plasma of healthy individuals (0.92 +/- 0.13 U/ml, 0.98 +/- 0.21 U/ml, 0.96 +/- 0.17, respectively). In a group of patients using oral anticoagulant therapy the ratio PS:Ca(II)Ag/PS:NonCa(II)Ag decreased at increasing intensity of anticoagulation suggesting the presence of sub- and noncarboxylated protein S molecules. In plasma of patients with a hereditary type I protein S deficiency PS:Ca(II)Ag and PS:NonCa(II)Ag were reduced to the same extent: mean ratio 1.02 +/- 0.12 in the group not on oral anticoagulant treatment and 0.94 +/- 0.10 in the group on oral anticoagulant therapy. Analysis of patients with a history of unexplained thrombo-embolic disease did not reveal individual patients with a PS:Ca(II)Ag/PS:NonCa(II)Ag ration below the lower limit of the normal range (mean ratio 1.05 +/- 0.17), suggesting that the frequency of genetic protein S variants with defects in the Ca(II)-stabilized conformation is very low.

Thrombomodulin activity in commercial thromboplastin preparations.

Recently, it was reported that endothelial cells contain a membrane protein which serves as a cofactor for the activation of protein C by thrombin (thrombomodulin). Because many commercial thromboplastins are prepared from vessel-rich tissues/organs (lung, brain, placenta), it was hypothesized that some preparations may contain significant amounts of thrombomodulin; theoretically, thrombomodulin contaminations may enhance the prothrombin time. Among 8 different commercial thromboplastins tested two thromboplastins were found to contain significant amounts of thrombomodulin activity: Simplastin (64 U/ml) and Ortho (12.8 U/ml). However, we were unable to demonstrate that the presence of the thrombomodulin activity in these thromboplastins could affect the prothrombin time either by delaying fibrin formation (formation of thrombin-thrombomodulin complexes) or by inhibiting activated factor V (formation of activated protein C).

The cofactor role of protein S in the acceleration of whole blood clot lysis by activated protein C in vitro.

The effect of purified human activated protein C (APC) and protein S on fibrinolysis was studied by using an in vitro blood clot lysis technique. Blood clots were formed from citrated blood (supplemented with 125I-fibrinogen) by adding thrombin and Ca2+-ions; lysis of the clots was achieved by adding tissue-type plasminogen activator. The release of labeled fibrin degradation products from the clots into the supernatant was followed in time. We clearly demonstrated that APC accelerates whole blood clot lysis in vitro. The effect of APC was completely quenched by antiprotein C IgG, pretreatment of APC with diisopropylfluorophosphate, and preincubation of the blood with antiprotein S IgG. This demonstrates that both the active site of APC and the presence of the cofactor, protein S, are essential for the expression of the profibrinolytic properties. At present, the substrate of APC involved in the regulation of fibrinolysis is not yet known. Analysis of the radiolabeled fibrin degradation products demonstrated that APC had no effect on the fibrin cross-linking capacity of factor XIII.