Publications by authors named "Andreas Weimann"

25 Publications

  • Page 1 of 1

Diagnostic value of plasma NGAL and intraoperative diuresis for AKI after major gynecological surgery in patients treated within an intraoperative goal-directed hemodynamic algorithm: A substudy of a randomized controlled trial.

Medicine (Baltimore) 2017 Jul;96(28):e7357

Department of Anesthesiology and Operative Intensive Care Medicine (CCM, CVK), Charité - Universitätsmedizin Berlin Labor Berlin-Charité Vivantes Services GmbH Department of Gynaecology, European Competence Center for Ovarian Cancer, Charité- University Medicine Berlin Charité-University Medicine Berlin and SOSTANA GmbH Berlin, Berlin, Germany.

Data on early markers for acute kidney injury (AKI) after noncardiovascular surgery are still limited. This study aimed to determine the diagnostic value of plasma neutrophil-gelatinase-associated lipocalin (pNGAL) and intraoperative diuresis for AKI in patients undergoing major abdominal surgery treated within a goal-directed hemodynamic algorithm.This study is a post-hoc analysis of a randomized controlled pilot trial comparing intravenous solutions within a hemodynamic goal-directed algorithm based on the esophageal Doppler in patients undergoing epithelial ovarian cancer surgery. The diagnostic value of plasma NGAL obtained at ICU admission and intraoperative diuresis was determined with respect to patients already meeting AKI criteria 6 hours after surgery (AKI6h) and to all patients meeting AKI criteria at least once during the postoperative course (AKItotal). AKI was diagnosed by the definition of the Kidney Disease Improving Global Outcome (KDIGO) group creatinine criteria and was screened up to postoperative day 3. Receiver operating characteristic curves including a gray zone approach were performed.A total of 48 patients were analyzed. None of the patients had increased creatinine levels before surgery and 14 patients (29.2%) developed AKI after surgery. Plasma NGAL was predictive for AKI6h (AUCAKI6h 0.832 (95% confidence interval [CI], 0.629-0.976), P = .001) and AKItotal (AUCAKItotal 0.710 (CI 0.511-0.878), P = .023). The gray zones of pNGAL calculated for AKI6h and AKItotal were 210 to 245 and 207 to 274 ng mL, respectively. The lower cutoffs of the gray zone at 207 and 210 ng mL had a negative predictive value (NPV) (i.e., no AKI during the postoperative course) of 96.8% (CI 90-100) and 87.1% (CI 78-97), respectively. Intraoperative diuresis was also predictive for AKI6h (AUCAKI6h 0.742 (CI 0.581-0.871), P = .019) with a gray zone of 0.5 to 2.0 mL kg h. At the lower cutoff of the gray zone at 0.5 mL kg h, corresponding to the oliguric threshold, the NPV was 84.2% (78-92).This study indicates that pNGAL can be used as an early marker to rule out AKI occurring within 3 days after major abdominal surgery. Intraoperative diuresis can be used to rule out AKI occurring up to 6 hours after surgery.

Trial Registration: ISRCTN 53154834.
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http://dx.doi.org/10.1097/MD.0000000000007357DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5515745PMC
July 2017

Delta-He, Ret-He and a New Diagnostic Plot for Differential Diagnosis and Therapy Monitoring of Patients Suffering from Various Disease-Specific Types of Anemia.

Clin Lab 2016 ;62(4):667-77

Background: The present study was aimed to prove the usefulness of a new diagnostic plot (Hema-Plot), illustrating the relationship between the hemoglobin content of reticulocytes (Ret-He) as a marker of functional iron deficiency and the difference between the reticulocyte and erythrocyte hemoglobin content (Delta-He) as a marker of an impaired hemoglobinization of newly formed reticulocytes occurring during inflammatory processes, to differentiate between various disease-specific types of anemia.

Methods: A complete blood and reticulocyte count was performed on routine EDTA blood samples from 345 patients with and without various disease-specific types of anemia using the Sysmex XN-9000 hematology analyzer: blood healthy newborns (n = 23), blood healthy adults (n = 31), patients suffering from anemia of chronic disease (ACD) due to diverse oncological, chronic inflammatory, or autoimmune diseases (total n = 138) with (n = 65) and without therapy (n = 73), patients with thalassemia and/or hemoglobinopathy (n = 18), patients with iron deficiency anemia (IDA) (n = 35), patients with a combination of ACD and IDA (n = 17), as well as patients suffering from sepsis (total n = 83) with (n = 32) and without therapy (n = 51). The results for Ret-He, Delta-He, and C-reactive protein (CRP) were statistically compared (Mann-Whitney U Test) between the particular patient groups and the diagnostic plots were drawn.

Results: Delta-Hemoglobin showed a statistically significant difference between blood healthy newborns and blood healthy adults (p ≤ 0.05), while Ret-He and C-reactive protein did not. In addition, of all three biomarkers only Delta-He showed a statistically significant difference (p ≤ 0.05) between the ACD/IDA and IDA cohort. Delta-He, Ret-He, and CRP showed a statistically significant difference between patient cohorts with and without therapy suffering from ACD, ACD/IDA, and sepsis before and after medical therapy (p ≤ 0.05). The Hema-Plot illustrated the dynamic character of Ret-He and Delta-He, notably in inflammation-based types of anemia like ACD or ACD/ IDA.

Conclusions: Delta-He is a new biomarker clearly distinguishing between inflammation-based types of anemia before and after medical therapy, as well as between ACD/IDA and IDA. The new Hema-Plot is a helpful tool for differential diagnosis and disease-monitoring in various types of disease-specific anemia, especially in ACD and ACD/IDA. The Hema-Plot can be used to identify non-adherent patients or an insufficient therapy.
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http://dx.doi.org/10.7754/clin.lab.2015.150830DOI Listing
June 2016

Nucleated red blood cells as marker for an increased risk of unfavorable outcome and mortality in very low birth weight infants.

Early Hum Dev 2015 Oct 25;91(10):559-63. Epub 2015 Jul 25.

Department of Neonatology, Charité-Universitätsmedizin Berlin, Germany.

Background: Nucleated red blood cells (NRBC) are normoblastic cells that failed to extrude their nuclei before exiting from bone marrow or liver. While NRBC are frequently found in umbilical cord blood after fetal distress, NRBC counts drop rapidly after birth.

Aims: To determine the predictive value of the NRBC count during the first 120h after birth as marker for later risk of unfavorable outcome in very preterm infants.

Study Design: This cohort study investigated the association between absolute count of NRBC on admission (day 1), the mean NRBC count between day 2 to 5, and outcome (mortality or the composite outcome of mortality and severe morbidity).

Results: 438 infants with a gestational age<32weeks and a birth weight<1500g were included within a five-year period, of whom 46 patients died and 65 suffered from severe morbidity. Nonsurvivors had significantly higher NRBC counts between day 2 and 5, as compared to survivors. This finding was observed in infants both appropriate and small for gestational age. An increase of 10/nL of the mean NRBC count on postnatal day 2 to 5 had an odds ratio for mortality of 6.95 (95% CI 2.21-21.86) and an odds ratio for the composite outcome mortality and severe morbidity of 3.43 (95% CI 1.43-8.24). The optimal cut-off value for prediction of death was NRBC >2/nL with a sensitivity of 85% and a specificity of 75%.

Conclusions: Elevation of NRBC on postnatal day 2 to 5 is an independent predictor of mortality in preterm infants.
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http://dx.doi.org/10.1016/j.earlhumdev.2015.06.004DOI Listing
October 2015

Detection and quantification of hypo- and hypergranulated neutrophils on the new Sysmex XN hematology analyzer: establishment of an adapted reference interval for the neutrophil-granularity-intensity compared to XE-technology in adult patients.

Clin Lab 2015 ;61(3-4):235-41

Background: Since the recent introduction of Sysmex hematology analyzers of the XN-series it can be expected that the values of individual hematological parameters might differ between the new XN and the well-established XE platform. One such parameter is called Neutrophil-Granularity-Intensity or NEUT-GI on the XN-series and NEUT-X on the XE-series. Both parameters are used by clinicians to calculate the Granularity-Index (GI-Index), an important tool to detect hypo- or hypergranulated neutrophils occurring during myelodysplasia or inflammation. The aims of this study were to determine if previously reported reference intervals for NEUT-X can be used for NEUT-GI as well and if the GI-Indices on both analyzer platforms correlate with each other.

Methods: NEUT-GI and NEUT-X were assessed in a set of 789 blood samples (n = 543 samples from adult intensive care units and n = 246 samples from adult "blood-healthy" control patients) and the corresponding Granularity-Indices were calculated for all samples using data obtained from XE-5000 and XN-1000 hematology analyzers.

Results: NEUT-GI and NEUT-X correlated significantly with each other (r2: 0.6512; p < 0.0001) with statistically significant higher values for NEUT-GI compared to NEUT-X in the control group (p < 0.0001) as well as in the ICU patients (p < 0.0001). This indicated that previously established reference intervals for NEUT-X cannot be used for NEUT-GI. In contrast, the GI-Indices showed no statistically significant difference between the analyzers in both groups. The GI-Indices were higher in the ICU patients compared to the control group on both analyzer platforms (p < 0.0001), as would be expected.

Conclusions: Our study revealed the emphatic need for a new reference interval for NEUT-GI on the XN platform. The resulting 95% reference intervals were 140.91 - 160.46 channels for NEUT-GI and 129.20 - 142.33 channels for NEUT-X. The GI-Indices showed no significant statistical difference between the XN- and XE-series in both cohorts.
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http://dx.doi.org/10.7754/clin.lab.2014.140704DOI Listing
June 2015

Intensive Care Infection Score--A new approach to distinguish between infectious and noninfectious processes in intensive care and medicosurgical patients.

J Int Med Res 2015 Jun 7;43(3):435-51. Epub 2015 Apr 7.

Department of Laboratory Medicine, Clinical Chemistry and Pathobiochemistry, Charité Campus Virchow-Klinikum, Charité - Universitätsmedizin Berlin, Berlin, Germany Labor Berlin - Charité Vivantes Services, Berlin, Germany.

Objectives: Clinicians regularly encounter substantial time delays in diagnosing sepsis and administering appropriate antibiotic treatment. This study investigated the ability of the Intensive Care Infection Score (ICIS) to distinguish between infectious and noninfectious processes, and to assess the justified commencement of antibiotic therapy retrospectively, in line with hospital actual best practice and applied laboratory parameters.

Methods: Intensive-care unit (ICU) patients were enrolled in this retrospective, observational study. Clinical data and laboratory parameters were determined daily. The cohort was divided into infected and noninfected patient groups.

Results: Out of 172 ICU patients, including 72 postoperative patients, the predictive value for infection throughout the first 5 days in 'all patients' and the 'postoperative patient' group was highest for ICIS. An ICIS cut-off value of three could predict infection in postoperative patients with 82.9% sensitivity and 75.1% specificity. ICIS showed the lowest rate of potentially 'falsely encouraged' and 'discouraged' antibiotic therapies for noninfected and for septic postoperative patients, respectively, compared with C-reactive protein, procalcitonin and white blood cell levels.

Conclusions: In the ICU, particularly for postoperative patients, ICIS is a reliable marker for the timely identification of infection. ICIS may qualify as a new decision support tool for antibiotic therapy, when interpreted within the clinical context.
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http://dx.doi.org/10.1177/0300060514557711DOI Listing
June 2015

Soluble CEACAM8 interacts with CEACAM1 inhibiting TLR2-triggered immune responses.

PLoS One 2014 17;9(4):e94106. Epub 2014 Apr 17.

ZIK Septomics, University Hospital Jena, Jena, Germany.

Lower respiratory tract bacterial infections are characterized by neutrophilic inflammation in the airways. The carcinoembryonic antigen-related cell adhesion molecule (CEACAM) 8 is expressed in and released by human granulocytes. Our study demonstrates that human granulocytes release CEACAM8 in response to bacterial DNA in a TLR9-dependent manner. Individuals with a high percentage of bronchial lavage fluid (BALF) granulocytes were more likely to have detectable levels of released CEACAM8 in the BALF than those with a normal granulocyte count. Soluble, recombinant CEACAM8-Fc binds to CEACAM1 expressed on human airway epithelium. Application of CEACAM8-Fc to CEACAM1-positive human pulmonary epithelial cells resulted in reduced TLR2-dependent inflammatory responses. These inhibitory effects were accompanied by tyrosine phosphorylation of the immunoreceptor tyrosine-based inhibitory motif (ITIM) of CEACAM1 and by recruitment of the phosphatase SHP-1, which could negatively regulate Toll-like receptor 2-dependent activation of the phosphatidylinositol 3-OH kinase-Akt kinase pathway. Our results suggest a new mechanism by which granulocytes reduce pro-inflammatory immune responses in human airways via secretion of CEACAM8 in neutrophil-driven bacterial infections.
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http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0094106PLOS
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3990526PMC
January 2015

Polymorphisms of the toll-like receptor 2 and 4 genes are associated with faster progression and a more severe course of sepsis in critically ill patients.

J Int Med Res 2014 Feb 23;42(1):93-110. Epub 2013 Dec 23.

Department of Anaesthesiology and Intensive Care Medicine, Charité - Universitätsmedizin Berlin, Campus Charité Mitte and Campus Virchow-Klinikum, Berlin, Germany.

Objective: To determine whether the Arg753Gln polymorphism of the toll-like receptor 2 (TLR2) gene and the Asp299Gly polymorphism of the TLR4 gene in critically ill patients affect their clinical outcomes.

Methods: Medical and surgical patients in three intensive care units (ICU) were enrolled in this prospective study. TLR2 and TLR4 gene polymorphisms were determined using restriction fragment length polymorphism analysis.

Results: A total of 145 patients were included in this study: 28 patients carried heterozygous mutations (10 in the TLR2 gene, 19 in the TLR4 gene, and one combined) and 117 patients were wild type. Severe sepsis was observed in 33% of wild types (n = 38), 60% of the TLR2 group (n = 6), and 63% of the TLR4 group (n = 12); the difference was significant between the TLR4 and wild type groups. Both TLR groups demonstrated a shorter time-to-onset of severe sepsis or septic shock. Only the TLR4 group demonstrated significant progression towards septic shock compared with the wild type group. Length of ICU stay was significantly prolonged in the TLR4 group compared with the wild type group, but not in the TLR2 group.

Conclusions: Two common SNPs of the TLR2 and TLR4 genes--Arg753Gln and Asp299Gly--were associated with a shorter time-to-onset of severe sepsis or septic shock in patients admitted to the ICU.
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http://dx.doi.org/10.1177/0300060513504358DOI Listing
February 2014

Profiling of Endo H-released serum N-glycans using CE-LIF and MALDI-TOF-MS--application to rheumatoid arthritis.

Electrophoresis 2011 Dec;32(24):3510-5

Glycodesign and Glycoanalytics, Institute of Laboratory Medicine, Clinical Chemistry and Pathobiochemistry, Charité Medical University, Berlin, Germany.

High-mannose and hybrid-type N-glycans are present in human serum glycoproteins in low abundance but have recently been described to play an important role in immune responses. It is therefore important to find a strategy to selectively analyze their structures in the context of health and disease in order to understand their impact on disease mechanisms. We report here the characterization of high-mannose and hybrid-type N-glycans in total human serum. To this end, N-glycans were released using Endo-β-N-acetylglucosaminidase H (Endo H) and analyzed by CE-LIF and MALDI-TOF-MS. We found that the high-mannose structures Man(5-9)GlcNAc(1) represented the majority of the pool. The monoglucosylated structure Glc(1)Man(9)GlcNAc(1) as well as four hybrid structures could be identified. Then, we compared the Endo H-released serum glycome of patients suffering from rheumatoid arthritis with healthy controls as mannose-binding lectin deficiency (MBL) and modulation of α-mannosidase activity were previously associated with this disease. Interestingly, we observed that both high-mannose and hybrid structures were fairly constant, suggesting that circulating MBL and α-mannosidase may not affect significantly the levels of serum glycoproteins carrying these glycans.
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http://dx.doi.org/10.1002/elps.201100250DOI Listing
December 2011

Immature platelet count: a simple parameter for distinguishing thrombocytopenia in pediatric acute lymphocytic leukemia from immune thrombocytopenia.

Pediatr Blood Cancer 2011 Oct 22;57(4):641-7. Epub 2010 Dec 22.

Department of Pediatrics, Charité - Universitätsmedizin Berlin, Berlin, Germany.

Background: Platelet counts below normal values define thrombocytopenia. However, platelet counts alone do not reveal the underlying pathomechanism. New blood cell counters provide additional information on platelet size and volume, and enable the distinction of sub-populations. In this preliminary study, we evaluate whether one of these markers can be used for diagnosis of isolated thrombocytopenia in children.

Procedure: We provide normal values for mean platelet volume (MPV), platelet distribution width (PDW), plateletcrit (PCT), platelet large cell ratio (P-LCR), platelet mean-frequent volume (P-MFV), relative immature platelet fraction (IPF%), and absolute IPF (IPF#) for 100 healthy children and analyzed 87 children with thrombocytopenia.

Results: In children with platelet production defects, IPF% was low, while in acute immune thrombocytopenia (ITP), IPF% was markedly increased (median 25.2%, P < 0.01), representing accelerated platelet turnover. Interestingly, children diagnosed with acute lymphocytic leukemia (ALL) also had elevated IPF% (median 10%, P < 0.01), suggesting that thrombopoiesis is stimulated despite virtual absence of bone marrow progenitors. Low IPF# was only found in patients with acute ITP.

Conclusions: IPF% is a marker for thrombocytopenia due to defective platelet production while IPF#, representing the immature platelet count, might become a practical parameter to distinguish acute ITP from thrombocytopenia in children with newly diagnosed ALL (P < 0.01).
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http://dx.doi.org/10.1002/pbc.22907DOI Listing
October 2011

Granularity Index of the SYSMEX XE-5000 hematology analyzer as a replacement for manual microscopy of toxic granulation neutrophils in patients with inflammatory diseases.

Clin Chem Lab Med 2011 Jul 17;49(7):1193-8. Epub 2011 May 17.

Institute of Laboratory Medicine and Pathobiochemistry, Charité Universitätsmedizin Berlin, Berlin, Germany.

Background: When certain inflammatory processes occur, toxic granulation neutrophils (TGNs) appear in the blood showing prominent cytoplasmic granules. Currently, the granularity of TGNs is analyzed by manual microscopy of blood smears. The SYSMEX XE-5000 is an automated hematology analyzer, which can measure toxic granulation of TGNs by calculating the Granularity (GI) Index. In this study we investigated if the GI-Index is suitable as a parameter for the TGN granularity in inflammatory diseases.

Methods: An evaluation of the toxic granulation neutrophil (TGN) granularity by manual microscopy, the GI-Index and the C-reactive protein (CRP) concentrations of 158 patients were determined. Blood samples from 40 healthy individuals were incubated with lipopolysaccharide (LPS) for in vitro kinetic measurements of the GI-Index. Furthermore, time course measurements of the GI-Index and CRP concentrations of 100 intensive care unit patients were performed.

Results: The GI-Index correlated with the microscopic rating of TGNs (n=158; r(s)=0.839; p<0.0001). When incubating the blood samples with LPS, the neutrophils displayed hypogranulation 30 min after incubation and a hypergranulation after 90 min. In vivo, the GI-Index indicated changes of the bacterial infection status 1 day earlier than the CRP concentration. The correlation of CRP and GI-Index varied between the patient cohorts (n=158; r(s)=0.836) (n=100; r=0.177), depending on the cause and extent of inflammation.

Conclusions: The GI-Index is suited to quantify the granularity of TGNs. The GI-Index is an automated, standardized parameter available on a 24 h basis. We suggest that it replace the time-consuming, subjective and semiquantitative microscopic procedure.
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http://dx.doi.org/10.1515/CCLM.2011.188DOI Listing
July 2011

In vitro cytotoxicity of the novel antimyeloma agents perifosine, bortezomib and lenalidomide against different cell lines.

Invest New Drugs 2012 Apr 16;30(2):480-9. Epub 2010 Nov 16.

Medizinische Klinik III, Hämatologie, Onkologie und Transfusionsmedizin, Charité Campus Benjamin Franklin, Hindenburgdamm 30, 12200 Berlin, Germany.

The novel AKT inhibitor perifosine, a synthetic alkylphospholipid, is currently being investigated in clinical trials for the treatment of different hematological and oncological malignancies. The in vitro cytotoxicity of perifosine, bortezomib and lenalidomide against 6 cell lines derived from hematological malignancies was investigated using trypan blue staining, flow cytometry-based detection of activated caspases, Annexin V assays, immunohistochemistry studies (KI-67 and caspase-3 staining) and the immature-myeloid-information (IMI) technique. Perifosine and bortezomib induced concentration- and time-dependent cytotoxicity in all cell lines tested. Perifosine together with bortezomib largely exerted additive or synergistic effects with combination indices ranging from 1.13 to 0.22 for combined efficacies of 25% to 75% after 24-hour incubation. Lenalidomide-triggered cytotoxicity was low in all cell lines tested with any assay (less than 10% compared to the negative control). Finally, perifosine, but not bortezomib or lenalidomide, significantly increased the number of cells detected in the IMI channel. Perifosine and bortezomib- but not lenalidomide- trigger substantial cytotoxicity by caspase activation and mainly act additively or synergistically. The IMI technique might be a useful tool for studying cytotoxicity of agents like perifosine that interact mainly with the cellular membrane.
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http://dx.doi.org/10.1007/s10637-010-9576-2DOI Listing
April 2012

Comparison of six immunohistochemical markers for the histologic diagnosis of neoplasia in Barrett's esophagus.

Virchows Arch 2010 Nov 16;457(5):537-45. Epub 2010 Sep 16.

Institute for Laboratory Medicine and Pathobiochemistry, Charité-University Medical Center, Berlin, Germany.

In esophageal neoplasms, the histopathologic differentiation between Barrett's esophagus with or without intraepithelial neoplasia and adenocarcinoma is often challenging. Immunohistochemistry might help to differentiate between these lesions. The expression of CDX2, LI-cadherin, mucin 2 (MUC2), blood group 8 (BG8, Lewis(y)), claudin-2, and villin was investigated in normal gastroesophageal (n = 23) and in Barrett's (n = 17) mucosa, in low-grade (n = 12) and high-grade (n = 9) intraepithelial neoplasia (IEN) as well as in esophageal adenocarcinoma (n = 16), using immunohistochemistry. For CDX2 and LI-cadherin, the immunoreactivity score was highest in IEN while for MUC2, BG8, and villin, it dropped gradually from Barrett's via IEN to adenocarcinoma, and expression of Claudin-2 was only weak and focal in all lesions. The expression of MUC2 and LI-cadherin differed significantly between all examined lesions except between low-grade and high-grade IEN. MUC2 and LI-cadherin are useful immunohistochemical markers for the differentiation between normal glandular mucosa, Barrett's mucosa, IEN, and invasive carcinoma of the esophagus; however, none of the examined markers was helpful for the differentiation between low-grade and high-grade IEN.
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http://dx.doi.org/10.1007/s00428-010-0972-yDOI Listing
November 2010

CDX2 and LI-cadherin expression in esophageal mucosa: use of both markers can facilitate the histologic diagnosis of Barrett's esophagus and carcinoma.

Int J Surg Pathol 2010 Oct 5;18(5):330-7. Epub 2010 May 5.

Charité University Medical Center, Berlin, Germany.

Background: Barrett's mucosa is a risk factor for esophageal adenocarcinoma and should be detected at an early stage. CDX2 and liver-intestine (LI)-cadherin are intestine-specific markers. Aberrant CDX2 expression has been demonstrated in Barrett's metaplasia, esophagitis, and intestinal metaplasia of the stomach.

Methods: The relationship between CDX2 and LI-cadherin expression was investigated in normal gastroesophageal (n = 24) and in Barrett's (n = 20) mucosa, in low-grade (n = 15) and high-grade (n = 13) intraepithelial neoplasia (IEN) as well as in esophageal adenocarcinoma (n = 16), using immunohistochemistry.

Results: Nuclear positivity for CDX2 coupled with membranous expression of LI-cadherin was observed in about 70% of the epithelial cells of Barrett's mucosa. The intensity of staining and the percentage of positive cells increased within the sequential steps of low-grade to high-grade IEN, whereas the normal cylindric epithelium lacked the expression of both. In adenocarcinoma, the expression of LI-cadherin and CDX2 was significantly weaker or absent.

Conclusions: CDX2 and LI-cadherin are sensitive markers of intestinal metaplasia with or without dysplasia in the upper gastrointestinal tract. Both can be helpful for the early histologic diagnosis of Barrett's esophagus and its subsequent lesions; however, they do not significantly discern between different grades of dysplasia.
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http://dx.doi.org/10.1177/1066896910364228DOI Listing
October 2010

Cardiac troponin T release and inflammation demonstrated in marathon runners.

Clin Lab 2010 ;56(1-2):51-8

Medizinische Klinik (Kardiologie), Charité--Universitätsmedizin Berlin, Berlin, Germany.

Background: As shown on the basis of highly sensitive assays, cardiac troponin release is now observed after physiological heart stress and in mild heart pathologies: both are [corrected] considered unrelated to the irreversible cardiac alteration that is typically the source of release. Transitory cardiac membrane leakage was suggested as the basis. In our view, mild inflammation may drive this type of cardiac troponin release. To verify this hypothesis, marathon runners who demonstrated post-run inflammation were used as a model to correlate cTnT release and inflammation intensity.

Methods: In 78 male marathon runners who participated in the BERLIN-MARATHON 2006, cardiac troponin T (cTnT) was monitored [corrected] at three time points (pre-race, post-race, and after two weeks of rest). [corrected] Measurements were done with the highly sensitive assay (hs cTnT assay) and the conventional fourth-generation cTnT assay for comparison. Concurrently, [corrected] the inflammation markers (leukocyte and neutrophil counts, CRP, IL-6) were measured.

Results: Pre-race, the fourth-generation assay failed to demonstrate cTnT positivity (> test specific LLD). In contrast, with the [corrected] use of the highly sensitive assay, 28% of the participants were positive for cTnT (> LLD of hs cTnT assay). Post-race, cTnT as measured with the fourth-generation assay was observed to be detectable in 43% of the runners (> LLD = 99(th) percentile cut off), but all runners had detectable cTnT values (> LLD) when measured with the highly sensitive assay. Even in 94% of these cTnT-positive runners, the value exceeded the 99(th) percentile cut off determined for the highly sensitive assay (13 ng/L). cTnT release correlated significantly with inflammation intensity. Faster runners demonstrated significantly stronger cTnT releases and inflammation signs.

Conclusions: As demonstrated after physiological heart stress such as marathon running, transitory inflammation is evidently one of the events contributing to the cardiac troponin release under conditions suggested as unrelated to irreversible cardiac alteration.
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April 2010

Immature platelet values indicate impaired megakaryopoietic activity in neonatal early-onset thrombocytopenia.

Thromb Haemost 2010 May 9;103(5):1016-21. Epub 2010 Mar 9.

Department of Neonatology, Charité - Universitätsmedizin Berlin, Charitéplatz 1, D-10117 Berlin, Germany.

Newly released platelets, referred to as immature platelets, can be reliably quantified based on their RNA content by flow cytometry in an automated blood analyser. The absolute number of immature platelets (IPF#) and the immature platelet fraction (IPF%) reflect megakaryopoietic activity. We aimed to analyse the implication of these parameters in analysing the pathomechanism of early-onset neonatal thrombocytopenia. Platelet counts and IPF were determined at day 1 to 3 (d1 to d3) in 857 neonates admitted to intensive care. In thrombocytopenic patients (platelet counts<150 x 10(9)/l, n=129), IPF# was significantly lower (8.5 +/- 2.7 x 10(9)/l), than in non-thrombocytopenic neonates (9.5 +/- 3.6 x 10(9)/l, n=682, p<0.05). IPF% was significantly higher in thrombocytopenic (9.3 +/- 7.9%) vs. non-thrombocytopenic neonates (4.1 +/- 1.8%, p<0.001). While neonates with early-onset infection (n=134) had lower platelet counts (199 +/- 75 x 10(9)/l) compared to controls (230 +/- 68 x 10(9)/l, n=574, p<0.01), there were no differences in IPF# or IPF%. Likewise, "small for gestational age" infants (SGA, n=149) had lower platelet counts at d1 (199 +/- 75 x 10(9)/l, p<0.001) than controls, but no differences in IPF. A trend towards lower IPF# was detected if SGA infants with platelet counts <100 x 10(9)/l (5.4 +/- 3.9 x 10(9)/l, n=11) and thrombocytopenic neonates with infection (9.9 +/- 7.3 x 10(9)/l, n=10, p=0.11) were compared. The evaluation of IPF# indicates that thrombocytopenia in neonates is likely due to a combination of increased platelet consumption and inadequate megakaryopoietic response by the neonatal bone marrow. Furthermore, SGA neonates with moderate and severe thrombocytopenia might have a pronounced suppression of megakaryopoiesis compared to neonates with infection.
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http://dx.doi.org/10.1160/TH09-03-0148DOI Listing
May 2010

Is there a role for mannan-binding lectin in the diagnosis of inflammatory bowel disease?

Immunogenetics 2010 Apr 25;62(4):231-5. Epub 2010 Feb 25.

Institute of Laboratory Medicine and Pathobiochemistry, Charité University Medicine Berlin, Augustenburger Platz 1, 13353, Berlin, Germany.

Mannan-binding lectin (MBL) activates the lectin-complement pathway as part of the innate immune defence by binding to the surface of microorganisms. Therefore, MBL2 presents an interesting candidate gene for the inflammatory bowel diseases, ulcerative colitis (UC) and Crohn's disease (CD). In our study, we evaluated the MBL serum concentrations and genotypes for diagnostic and classification purposes of patients with CD and UC. The MBL serum concentration was analysed in 98 CD patients and in 83 UC patients. In total, 82 patients with inflammatory rheumatic disorders and 189 healthy individuals served as controls. All study subjects were genotyped for the MBL2 polymorphisms G54D, G57E and R52C and the NOD2 (CARD15) mutations R702W, G908R and L1007fsinsC. Neither the median MBL serum concentration nor the MBL2 genotype distribution differed significantly between cohorts. Measurement of MBL serum concentrations offers no benefit for the diagnosis of CD or UC.
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http://dx.doi.org/10.1007/s00251-010-0429-0DOI Listing
April 2010

Choice of plasmid preparation kit influences sequencing results.

Biotechnol Lett 2010 Mar 7;32(3):421-2. Epub 2009 Nov 7.

Institute of Laboratory Medicine and Pathobiochemistry, Charité University Medicine Berlin, Augustenburger Platz 1, 13353, Berlin, Germany.

Commercially available plasmid preparation kits are used in many laboratories to isolate plasmid DNA for subsequent dideoxy sequencing. We tested kits from seven different manufacturers for the purity of obtained plasmid preparations and compared the quality of sequencing results. The amount of RNA carry-over differed significantly (80-323 ng) and the usage of two kits led to protein remainders. Sequence read lengths (0-812 bases) and percentages of reading errors (1.6-100%) were also significantly different. We found the amount of protein contaminants to correlate with the percentage of reading errors (r = 0.63, P = 0.02).
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http://dx.doi.org/10.1007/s10529-009-0157-4DOI Listing
March 2010

Boy with pseudohypoparathyroidism type 1a caused by GNAS gene mutation (deltaN377), Crouzon-like craniosynostosis, and severe trauma-induced bleeding.

Am J Med Genet A 2009 Jul;149A(7):1487-93

Institute of Medical Genetics, Charité Universitätsmedizin Berlin, Berlin, Germany.

We report on a 6-month-old boy with craniosynostosis, pseudohypoparathyroidism type 1a (PHP1A), and a GNAS gene mutation. He had synostoses of the coronal, frontal, and sagittal sutures, brachyturricephaly, and hydrocephaly. He also had congenital hypothyroidism, round face, full cheeks, shortness of limbs, mild developmental delay, and muscular hypotonia. Because of progressive hydrocephaly, the synostosis was corrected surgically but circulatory decompensation led to disseminated intravascular coagulation and cerebral infarctions. Our patient died 8 days later. Postmortem molecular studies of GNAS, the gene for guanine nucleotide-binding protein, alpha-stimulating activity polypeptide (gene for PHP1A), identified a de novo heterozygous 3 bp in frame deletion predicting a deletion of the asparagine residue at position 377 (deltaN377). This is the second report of this mutation. Results of molecular studies of craniosynostosis genes (FGFR2, FGFR3) and of numerous genetic variants predisposing to bleeding disorders were normal. We question whether craniosynostosis and trauma-induced bleeding disorder may be manifestations of PHP1A, or if our patient had two or three different congenital disorders.
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http://dx.doi.org/10.1002/ajmg.a.32889DOI Listing
July 2009

[Laboratory haematological changes in the field of intensive care medicine--the extended differential blood count].

Anasthesiol Intensivmed Notfallmed Schmerzther 2009 Mar 5;44(3):164-70; quiz 173. Epub 2009 Mar 5.

One of the most frequently ordered tests by intensive care medical staff is the complete (CBC) and differential blood count for diagnosing anemia due to blood loss, characterizing thrombopenic conditions or finding proof for sepsis. Classically, the clinician relies on the quantification of single cell classes for substitutive therapy especially in emergency situations. Here, the authors discuss new possibilities offered by markers of the extended blood count from fully automatized haematology analyzers supplying valuable information for monitoring therapies in intensive care units.
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http://dx.doi.org/10.1055/s-0029-1215546DOI Listing
March 2009

Mayo risk score for primary biliary cirrhosis: a useful tool for the prediction of course after liver transplantation?

Ann Transplant 2008 ;13(3):35-42

Department of General, Visceral and Transplantation Surgery, Humboldt University of Berlin, Charité Virchow Clinic, Berlin, Germany.

Background: Survival after orthotopic liver transplantation (OLT) for primary biliary cirrhosis (PBC) is excellent. In order to define the optimal time point for OLT, the Mayo risk score (MRS) was developed and a score of 7.8 was identified for transplantation. However, in reality most recipients are in a bad condition with a MRS above 7.8. So far it is still unknown if a higher score is associated with more complications after OLT perioperatively and in a long-term follow-up. Therefore, this study was designed to investigate the association of the MRS score with postoperative survival and complications.

Material/methods: Between 1989 and 2006, 115 patients were transplanted for histologically proven PBC at the Charité Campus Virchow Clinic. In 98 of these patients, MRS data was available and retrospectively analyzed. Median age of 87 women and 11 men was 54 years (25 to 66 years).

Results: The median follow-up after liver transplantation was 109 months (0.5-205 months). Actuarial patient survival after 5, 10 and 15 years was 90%, 88%, and 83%. Calculated survival by MRS without transplantation after 1, 5 and 7 years was 20%, 2% and 1% for these patients. Twelve patients (12%) died and histological recurrence of PBC was detected in 14 patients (14%). Seven patients underwent retransplantation (7%) and 58 patients developed an acute rejection episode (59%). Mean MRS was in all patients 9.54+/-1.35 and did not differ between patients with lethal course, retransplantations, PBC recurrence, rejection episodes and duration of hospital stay. Classification of all patients into a low (<8.5), middle (8.5-10) and high MRS score (>10) did not show a significant difference in long-term survival. Univariate analysis for identifying the level of MRS as risk for death, PBC-recurrence, retransplantation, acute rejection episodes and hospital stay only showed a significant increased risk for acute rejection episodes (1 episode = 0.04; 2 episodes = 0.01) for patients with a MRS above 8.5.

Conclusions: The Mayo risk score is an approved mathematical model predicting survival in non-transplanted patients suffering from PBC. However, the score did not predict the course of our liver transplanted patients in a long-term follow-up. We could not demonstrate a reduced patient survival at a median MRS of 9.4 and about 10.0. Therefore, it is, from our point of view, questionable if the optimal time point for OLT is still 7.8.
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January 2009

Lithium--a role in hyperlipidemia?

Metabolism 2007 Apr;56(4):583-5

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http://dx.doi.org/10.1016/j.metabol.2006.11.001DOI Listing
April 2007

Novel two-stage screening procedure leads to the identification of a new class of transfection enhancers.

J Gene Med 2006 Jun;8(6):745-53

Institute of Microbiology and Genetics, Technical University Berlin, Gustav-Meyer-Allee 25, D-13355 Berlin.

Background: Non-viral gene transfer efficiency is low as compared to viral vector systems. Here we describe the discovery of new drugs that are capable of enhancing non-viral gene transfer into mammalian cells using a novel two-stage screening procedure.

Methods: First, potential candidates are preselected from a molecular library at various concentrations by a semi-automated yeast transfection screen (YTS). The maximal transfection efficiency of every positive drug is subsequently determined in independent experiments at the optimal concentration and compared to the inhibitory effect of the drug on cell growth (IC50). In a subsequent mammalian cell transfection screen (MTS), the maximal transfection efficiency and the IC50 are determined for all preselected drugs using a human cell line and a luciferase reporter gene construct.

Results: Employing our novel system we have been able to identify a new class of transfection enhancers, the tricyclic antidepressants (i.e. doxepin, maprotiline, desipramine and amoxapine). All positive drugs enhanced gene transfer in both yeast and human cell lines, but lower concentrations were sufficient for mammalian cells. With a triple combination of doxepin, amoxapine and chloroquine we obtained a transfection efficiency that exceeded that of chloroquine, one of the best-known transfection enhancers of mammalian cells, by nearly one order of magnitude.

Conclusions: Non-viral gene transfer efficiency can be increased significantly using new transfection enhancers that are identified by a novel, semi-automated two-stage screening system employing yeast cells in the first and specific human target cells in the second round.
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http://dx.doi.org/10.1002/jgm.887DOI Listing
June 2006

Desalting minimal amounts of DNA for electroporation in E. coli: a comparison of different physical methods.

Biotechnol Lett 2005 Jul;27(14):1003-5

Institut für Laboratoriumsmedizin und Pathobiochemie, Charité Universitätsmedizin Berlin, Campus Virchow-Klinikum Augustenburger Platz 1, D-13353, Berlin, Germany.

Recovery and purification of ligated plasmid DNA is a critical and limiting step in modern molecular cloning techniques prior to electroporation in E. coli. Four different DNA purification and desalting methods were compared in regard to simple handling, reproducibility, efficiency and cost-effectiveness using picogram and nanogram amounts of DNA. Microcolumn purification of DNA was up to two orders of magnitude more efficient compared to gel filtration, ethanol precipitation or drop dialysis.
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http://dx.doi.org/10.1007/s10529-005-7867-zDOI Listing
July 2005