Medical Laboratory Bremen

Bremen, Germany

Medical Laboratory Bremen

Bremen, Germany
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Heitland P.,Medical Laboratory Bremen | Blohm M.,University of Hamburg | Breuer C.,Kinderarzte an der Rathausallee | Brinkert F.,University of Hamburg | And 3 more authors.
Journal of Trace Elements in Medicine and Biology | Year: 2017

ICP-MS and HPLC-ICP–MS were applied for diagnosis and therapeutic monitoring in a severe intoxication with a liquid containing hexavalent chromium (Cr(VI)) and inorganic arsenic (iAs). In this rare case a liver transplantation of was considered as the only chance of survival. We developed and applied methods for the determination of Cr(VI) in erythrocytes and total chromium (Cr) and arsenic (As) in blood, plasma, urine and liver tissue by ICP-MS. Exposure to iAs was diagnosed by determination of iAs species and their metabolites in urine by anion exchange HPLC-ICP–MS. Three days after ingestion of the liquid the total Cr concentrations were 2180 and 1070 μg/L in whole blood and plasma, respectively, and 4540 μg/L Cr(VI) in erythrocytes. The arsenic concentration in blood was 206 μg/L. The urinary As species concentrations were <0.5, 109, 115, 154 and 126 μg/L for arsenobetaine, As(III), As(V), methylarsonate (V) and dimethylarsinate (V), respectively. Total Cr and As concentrations in the explanted liver were 11.7 and 0.9 mg/kg, respectively. Further analytical results of this case study are tabulated and provide valuable data for physicians and toxicologists. © 2017

Wolff D.,Medical Laboratory Bremen | Gerritzen A.,Medical Laboratory Bremen
Clinical Laboratory | Year: 2017

Background: A rapid molecular test for identifying cases of infectious diarrhea might be beneficial for patients. Methods: We established a LightMix modular multiplex PCR assay for rapid detection of Campylobacter, Salmonella., Yersinia, and Shigella species from fecal samples. Results: Unlike Campylobacter sp., direct detection of Salmonella from feces by multiplex PCR was significantly less sensitive than culture. Only 11 out of 21 (52%) Salmonella culture positive specimens were positive for Salmonella in the direct multiplex PCR assay (Campylobacter: 52/53 samples (98%]). In contrast, an overnight selenite enrichment step prior to multiplex PCR increased the Salmonella assay sensitivity significantly: 18 out of 18 Salmonella culture positive samples were then also detected by PCR. Conclusions: An overnight enrichment step is necessary for reliable PCR detection of Salmonella from fecal samples.

Rakers F.,Jena University Hospital | Bischoff S.,Jena University Hospital | Schiffner R.,Jena University Hospital | Haase M.,Jena University Hospital | And 8 more authors.
American Journal of Obstetrics and Gynecology | Year: 2015

Objective We sought to evaluate whether in addition to cortisol, catecholamines also transfer psychosocial stress indirectly to the fetus by decreasing uterine blood flow (UBF) and increasing fetal anaerobic metabolism and stress hormones. Study Design Seven pregnant sheep chronically instrumented with uterine ultrasound flow probes and catheters at 0.77 gestation underwent 2 hours of psychosocial stress by isolation. We used adrenergic blockade with labetalol to examine whether decreased UBF is catecholamine mediated and to determine to what extent stress transfer from mother to fetus is catecholamine dependent. Results Stress induced transient increases in maternal cortisol and norepinephrine (NE). Maximum fetal plasma cortisol concentrations were 8.1 ± 2.1% of those in the mother suggesting its maternal origin. In parallel to the maternal NE increase, UBF decreased by maximum 22% for 30 minutes (P <.05). Fetal NE remained elevated for >2 hours accompanied by a prolonged blood pressure increase (P <.05). Fetuses developed a delayed and prolonged shift toward anaerobic metabolism in the presence of an unaltered oxygen supply. Adrenergic blockade prevented the stress-induced UBF decrease and, consequently, the fetal NE and blood pressure increase and the shift toward anaerobic metabolism. Conclusion We conclude that catecholamine-induced decrease of UBF is a mechanism of maternal-fetal stress transfer. It may explain the influence of maternal stress on fetal development and on programming of adverse health outcomes in later life especially during early pregnancy when fetal glucocorticoid receptor expression is limited. © 2015 Elsevier Inc.

Pavlova A.,University of Bonn | Preisler B.,University of Bonn | Driesen J.,University of Bonn | de Moerloose P.,University of Geneva | And 5 more authors.
Haemophilia | Year: 2015

Combined coagulation factor VII (FVII) and factor X (FX) deficiency (combined FVII/FX deficiency) belongs to the group of bleeding disorders in which both factors show reduced plasma activity. It may arise from coincidental inheritance of separate coagulation factor deficiencies or a common cause as large deletions comprising both gene loci. The F7 and F10 genes are located on the long arm of chromosome 13. Here, we describe 10 cases with combined FVII/FX deficiency representing both genetic mechanisms of occurrence. Genetic analyses included direct sequencing of the F7 and F10 genes and MLPA (multiplex ligation-dependent probe amplification) for detection of heterozygous large deletions. In four patients, the combined deficiency was due to a large deletion within the terminal end of chromosome 13. In the remaining six cases the deficiency resulted from coincidental inheritance of different genetic alterations affecting both genes independently. In most cases, the genetic defects were heterozygous, presenting with prolonged PT, normal aPTT and mild or no bleeding symptoms. Only in one case compound heterozygous mutations were detected in the F10, resulting in prolonged aPTT and a more severe bleeding phenotype. To avoid a misdiagnosis of combined FVII/FX deficiency, analyses of single factor activities have to be performed in all cases with prolonged PT even if aPTT is normal. Genetic analyses are substantial for correct prediction of an inheritance pattern and a proper genetic counselling. © 2014 John Wiley & Sons Ltd.

Roehl A.B.,RWTH Aachen | Zoremba N.,RWTH Aachen | Kipp M.,RWTH Aachen | Schiefer J.,RWTH Aachen | And 6 more authors.
BMC Neurology | Year: 2012

Backround: Neuroprotective strategies after cardiopulmonary resuscitation are currently the focus of experimental and clinical research. Levosimendan has been proposed as a promising drug candidate because of its cardioprotective properties, improved haemodynamic effects in vivo and reduced traumatic brain injury in vitro. The effects of levosimendan on brain metabolism during and after ischaemia/hypoxia are unknown.Methods: Transient cerebral ischaemia/hypoxia was induced in 30 male Wistar rats by bilateral common carotid artery clamping for 15 min and concomitant ventilation with 6% O2 during general anaesthesia with urethane. After 10 min of global ischaemia/hypoxia, the rats were treated with an i.v. bolus of 24 μg kg-1 levosimendan followed by a continuous infusion of 0.2 μg kg-1 min-1. The changes in the energy-related metabolites lactate, the lactate/pyruvate ratio, glucose and glutamate were monitored by microdialysis. In addition, the effects on global haemodynamics, cerebral perfusion and autoregulation, oedema and expression of proinflammatory genes in the neocortex were assessed.Results: Levosimendan reduced blood pressure during initial reperfusion (72 ± 14 vs. 109 ± 2 mmHg, p = 0.03) and delayed flow maximum by 5 minutes (p = 0.002). Whereas no effects on time course of lactate, glucose, pyruvate and glutamate concentrations in the dialysate could be observed, the lactate/pyruvate ratio during initial reperfusion (144 ± 31 vs. 77 ± 8, p = 0.017) and the glutamate release during 90 minutes of reperfusion (75 ± 19 vs. 24 ± 28 μmol·L-1) were higher in the levosimendan group. The increased expression of IL-6, IL-1ß TNFα and ICAM-1, extend of cerebral edema and cerebral autoregulation was not influenced by levosimendan.Conclusion: Although levosimendan has neuroprotective actions in vitro and on the spinal cord in vivo and has been shown to cross the blood-brain barrier, the present results showed that levosimendan did not reduce the initial neuronal injury after transient ischaemia/hypoxia. © 2012 Roehl et al.; licensee BioMed Central Ltd.

PubMed | Medical Laboratory Bremen and German Environment Agency Umweltbundesamt
Type: Journal Article | Journal: International journal of hygiene and environmental health | Year: 2016

The broadband herbicide glyphosate (N-[phosphonomethyl]-glycine) and its main metabolite aminomethylphosphonic acid (AMPA) were analyzed by GC-MS-MS in 24h-urine samples cryo-archived by the German Environmental Specimen Bank (ESB). Samples collected in 2001, 2003, 2005, 2007, 2009, 2011, 2012, 2013, 2014, and 2015 were chosen for this retrospective analysis. All urine samples had been provided by 20 to 29 years old individuals living in Greifswald, a city in north-eastern Germany. Out of the 399 analyzed urine samples, 127 (=31.8%) contained glyphosate concentrations at or above the limit of quantification (LOQ) of 0.1g/L. For AMPA this was the case for 160 (=40.1%) samples. The fraction of glyphosate levels at or above LOQ peaked in 2012 (57.5%) and 2013 (56.4%) after having discontinuously increased from 10.0% in 2001. Quantification rates were lower again in 2014 and 2015 with 32.5% and 40.0%, respectively. The overall trend for quantifiable AMPA levels was similar. Glyphosate and AMPA concentrations in urine were statistically significantly correlated (spearman rank correlation coefficient=0.506, p0.001). Urinary glyphosate and AMPA levels tended to be higher in males. The possible reduction in exposure since 2013 indicated by ESB data may be due to changes in glyphosate application in agricultural practice. The ESB will continue monitoring internal exposures to glyphosate and AMPA for following up the time trend, elucidating inter-individual differences, and contributing to the ongoing debate on the further regulation of glyphosate-based pesticides.

Clajus C.,Hannover Medical School | Kuhn-Velten W.N.,Medical Laboratory Bremen | Schmidt J.J.,Hannover Medical School | Lorenzen J.M.,Hannover Medical School | And 3 more authors.
BMC Pharmacology and Toxicology | Year: 2013

Background: Dosing of antibiotics in critically ill patients is challenging. It becomes even more difficult if renal or hepatic impairment ensue. Modern means of renal replacement therapy are capable of removing antibiotics to a higher rate than decades ago, leaving clinicians with a high degree of uncertainty concerning the dose of antibiotics in this patient population. Cotrimoxazole, a combination of trimethoprim (TMP) and sulfamethoxazole (SMX) is frequently used in the treatment of several infections including Pneumocystis jirovecii pneumonia (PCP).Case presentation: Here we describe a patient with acute kidney injury in which we investigated the TMP and SMX levels during the course of an ICU stay. Cotrimoxazole was administered every six hours i.v. in a dose of TMP/SMX 15/75 mg/kg/day. Extended dialysis was performed with a high-flux dialyzer. Blood samples, as well as pre- and postdialyzer samples and aliquots of the collected spent dialysate were collected.Observed peak concentrations (Cmax) were 7.51 mg/l for TMP and 80.80 mg/l for SMX. Decline of blood levels during extended dialysis (TMP 64%; SMX 84%) was mainly due to removal by the dialysis procedure, illustrated by the high dialyzer clearances (median of 4 extended dialysis sessions: TMP 94.0 / SMX 51.0 ml/min), as well as by the absolute amount of both substances in the collected spent dialysate (median of 6 extended dialysis sessions: TMP 556 mg / SMX 130 mg). Within the limitation of a case report our data from 4 consecutive extended dialysis sessions suggest that this procedure substantially removes both TMP and SMX.Conclusions: Dose reduction, which is usually advocated in patients with acute kidney injury under renal replacement therapy, might lead to significant under-dosing. Pharmacokinetic studies for TMP/SMX dosing in this patient population are necessary to allow adequate dosing. © 2013 Clajus et al.; licensee BioMed Central Ltd.

Background: Testing of saved Ixodes ticks for the presence of Borrelia burgdorferi after biting a patient may be helpful to assess the risk of bacterial transmission and to decide upon preventive antibiotic treatment. A commercially available real-time PCR assay for the identification and quantification of Borrelia burdorferi sensu lato species (LightMix Kit for the detection of Borrelia spp., TIB MOLBIOL) was evaluated. Methods: Clinical and laboratory specimens were analysed for Borrelia DNA, including seven official proficiency panels (INSTAND, Germany, 2004 to 2009)), kit standards and 1121 ticks saved from bitten patients from all over Germany. Results: Borrelia burgdorferi sensu stricto, Borrelia afzelii, Borrelia garinii, Borrelia spielmanii, and Borrelia valaisiana were reliably detected by the real-time PCR. The recently described Borrelia lusitaniae (not pathogenic for humans) was not detected. Melt curve analysis of the PCR products allowed for a reliable Borrelia genospecies differentiation. Dilution series of proficiency panel samples revealed a detection limit of approximately 10 1 genomes / mL. Based on CP values the mean interassay coefficient of variation of the assay was 3.6 %, while the intraassay coefficients of variation were 1.6 % and 0.8 % for a strong positive and a weak positive kit standard, respectively. 20.2 % of the clinical ticks revealed detectable Borrelia burgdorferi DNA, with Borrelia afzelii being the most prominent subspecies. The quantification of Borrelia genomes in individual ticks showed a broad distribution ranging from 0.5 × 10 1 to 2.6 × 10 7 Borrelia equivalents per tick (median 1.6 × 10 4). Conclusions: The LightMix Kit for the detection of Borrelia spp. represents an excellent tool for the identification and quantification of Borrelia species in ticks. It can serve to further elucidate the relevance of preventive antibiotic treatment following tick bites in upcoming clinical studies.

Wolff D.,Medical Laboratory Bremen | Gerritzen A.,Medical Laboratory Bremen
Clinical Laboratory | Year: 2011

Background: The recent identification of murine leukemia virus (MLV)-related viruses in patients with chronic fatigue syndrome (CFS) has aroused much interest, not least among sufferers. However, other studies failed to detect these viruses in CFS patients. Methods: We wanted to establish a MLV-related virus real-time PCR for routine diagnostics. Results: Our study identified false positive MLV-related virus results due to a contamination of Superscript III Platinum One-Step Quantitative RT-PCR System (Invitrogen). Conclusions: This observation may be helpful to elucidate discrepant results for the detection of MLV-related virus like xenotropic MLV-related virus (XMRV) in recently published studies.

Gerritzen A.,Medical Laboratory Bremen | Wittke J.-W.,Medical Laboratory Bremen | Wolff D.,Medical Laboratory Bremen
Clinical Laboratory | Year: 2011

Background: From May until July 2011 a large outbreak of infections with Shiga toxin (Stx)-producing Escherichia coli (STEC) occurred in Germany. More than 800 patients suffered from hemolytic uremic syndrome (HUS), and 49 fatal cases were reported. Obviously, a mandatory requirement for such a clinical situation is the availability of rapid and reliable STEC tests from the investigating laboratory. The standard methods like enzyme immunoassay (EIA), vero cell cytotoxicity assay (VCA), and microbiological culture are, however, hampered by a lack of sensitivity and specificity unless a prior, time consuming broth enrichment step is employed. In order to acelerate the laboratory diagnosis, we evaluated an in-house real-time PCR assay for the detection of the Stx genes (stxl and stx2) directly from stool specimens without the need of broth enrichment procedures. Methods: 754 faecal samples were collected from 481 predominantly hospitalised patients with diarrhea from May 23 to June 10, 2011 at the Medical Laboratory Bremen, Germany. The samples were analysed with a direct stx real-time PCR and compared to EIA, VCA and culturing on enterohemolysin, ESBL, and CPS agar after broth enrichment. In addition, artificial samples (n = 12) from three official EHEC/STEC PCR quality proficiency panels (INSTAND, Germany, September 2006, September 2007, and April 2008) were analysed by real-time PCR only. Results: The real-time PCR produced reliable, distinct melting profiles with characteristic peaks for the stxl and stx2 PCR products. The quality proficiency panels revealed a detection limit of 10 CFU/PCR per reaction. 112, 86, 99, and 122 of 754 clinical samples were positive for culture, EIA, VCA, and real-time PCR, respectively. 121 of 122 PCR samples were positive only for stx2. Compared to culture as the gold standard, sensitivities of EIA, VCA, and real-time PCR were 76.8%, 83.9%, and 96.4% and specificities were 99.4%, 99.2%, and 97.8%, respectively. Conclusions: The direct fecal stx real-time PCR proved superior to enrichment based VCA and EIA and can be recommended as a quick and sensitive tool for the early diagnosis of STEC infection in addition to microbiological culture.

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