Anne Schlabrakowski

ChAT and NOS in human myenteric neurons: co-existence and co-absence

Most myenteric neurons contain one of the two generating enzymes for major excitatory and inhibitory neurotransmitters: choline acetyltransferase (ChAT) or neuronal nitric oxide synthase (NOS). Two minor groups of myenteric neurons contain either both enzymes or neither. Our study had two aims: (1) to compare the proportions of neurons stained for ChAT and/or NOS in human small and large intestinal whole-mounts by co-staining with an antibody against the human neuronal protein Hu C/D (HU); (2) to characterize these neurons morphologically by co-staining with a neurofilament (NF) antibody. In small intestinal whole-mounts co-stained with HU, we counted more ChAT-positive (ChAT+) than NOS+ neurons (52% vs. 38%), whereas the large intestine exhibited fewer ChAT+ than NOS+ neurons (38% vs. 50%). Neurons co-reactive for both ChAT and NOS accounted for about 3% in both regions, whereas neurons negative for both enzymes accounted for 7% in the small intestine and 8% in the large intestine. Co-staining with NF revealed that, in both small and large intestine, ChAT+/NOS+ neurons were either spiny (type I) neurons or displayed smaller perikarya that were weakly or not NF-stained. Of all spiny neurons, almost one third was co-reactive for ChAT and NOS, whereas nearly two thirds were positive only for NOS. Neurons negative for both ChAT and NOS were heterogeneous in size and NF reactivity. Thus, neither the co-existence nor the co-absence of ChAT and NOS in human myenteric neurons is indicative for particular neuron types, with several qualitative and quantitative parameters showing a wide range of interindividual variability.

One Step Nucleic Acid Amplification (OSNA) - a new method for lymph node staging in colorectal carcinomas

BackgroundAccurate histopathological evaluation of resected lymph nodes (LN) is essential for the reliable staging of colorectal carcinomas (CRC). With conventional sectioning and staining techniques usually only parts of the LN are examined which might lead to incorrect tumor staging. A molecular method called OSNA (One Step Nucleic Acid Amplification) may be suitable to determine the metastatic status of the complete LN and therefore improve staging.MethodsOSNA is based on a short homogenisation step and subsequent automated amplification of cytokeratin 19 (CK19) mRNA directly from the sample lysate, with result available in 30-40 minutes. In this study 184 frozen LN from 184 patients with CRC were investigated by both OSNA and histology (Haematoxylin & Eosin staining and CK19 immunohistochemistry), with half of the LN used for each method. Samples with discordant results were further analysed by RT-PCR for CK19 and carcinoembryonic antigen (CEA).ResultsThe concordance rate between histology and OSNA was 95.7%. Three LN were histology+/OSNA- and 5 LN histology-/OSNA+. RT-PCR supported the OSNA result in 3 discordant cases, suggesting that metastases were exclusively located in either the tissue analysed by OSNA or the tissue used for histology. If these samples were excluded the concordance was 97.2%, the sensitivity 94.9%, and the specificity 97.9%. Three patients (3%) staged as UICC I or II by routine histopathology were upstaged as LN positive by OSNA. One of these patients developed distant metastases (DMS) during follow up.ConclusionOSNA is a new and reliable method for molecular staging of lymphatic metastases in CRC and enables the examination of whole LN. It can be applied as a rapid diagnostic tool to estimate tumour involvement in LN during the staging of CRC.

Microsomal triglyceride transfer protein polymorphism (−493G/T) is associated with hepatic steatosis in patients with chronic hepatitis C

Background: Hepatic steatosis may promote progression of chronic hepatitis C (CHC). Microsomal triglyceride transfer protein (MTP) is required for assembly and secretion of ApoB lipoprotein and is implicated in hepatitis C virus (HCV)‐related steatosis. The MTP −493G/T polymorphism may promote liver fat accumulation, but its role in HCV‐related steatosis is still unclear.

Gastric cancer patients less than 50 years of age exhibit significant downregulation of E-cadherin and CDX2 compared to older reference populations

PURPOSE There is an increasing need to identify molecular markers, which can be used to prognosticate patient populations in gastric cancer. Whereas a significant number have been identified, very few have been characterized in the context of their ability to discriminate between young and old age groups in which a survival difference clearly exists. MATERIAL/METHODS In this study, using immunohistochemistry, we evaluated three markers with proven involvement in gastric cancer. The p53 tumor suppressor, the cell adhesion glycoprotein epithelial cadherin (CDH1) and the caudal-related homeobox transcription factor (CDX2) all of these have important roles in the aetiopathogenesis and/or progression of gastric cancer. RESULTS After adjustments for TNM stage, tumor grade, histopathological characteristics (Lauren classification), we found significant differences in the expression of these proteins, particularly E-cadherin and CDX2 between young and elderly patients. However, these differences did not amount to a significant difference in survival. CONCLUSIONS This study demonstrates that the protein expression of p53, CDH1 and CDX2 significantly discriminates young patients with gastric cancer who have a better prognostic outlook from older patients, but this difference in expression does not contribute to a survival benefit.

COX-2, TFF1, and Src define better prognosis in young patients with gastric cancer.

Despite its dwindling occurrence, gastric cancer remains a leading cause of cancer related mortality worldwide. Molecular determinants of prognosis that impact survival are being sought out as a means to facilitate rational clinical decision‐making and enhance patient management. In this study, we evaluated three molecules implicated in gastric carcinogenesis and demonstrated that the differential expression of cyclooxygenase‐2 (COX‐2) and the viral oncogene homolog Src proteins could explain the differences in survival observed in patients older and younger than 50 years of age.

Quantitative Estimation and Chemical Coding of Spiny Type I Neurons in Human Intestines

Previous studies have shown that most human myenteric neurons co-staining for vasoactive intestinal peptide (VIP), neuronal nitric oxide synthase (nNOS) and neurofilaments (NF) display the morphology of spiny type I neurons displaying a descending projection pattern. Here, we estimated the proportions of spiny neurons in human intestines, the amount of congruence of VIP/nNOS-immunoreactive with spiny neurons and whether galanin (GAL) is co-localized with VIP. Three sets of colchicine-pretreated and fixed whole mounts of 21 patients or body donors (median age 65 years; 10 female, 11 male) were stained for VIP, nNOS and NF, for VIP, nNOS and the human neuronal protein Hu C/D (HU) as well as for VIP, nNOS and GAL. The majority of VIP/nNOS-co-reactive neurons were spiny neurons (79/80% in small/large intestine, respectively) and the majority of spiny neurons co-stained for VIP and nNOS (82/69%). Neurons co-immunoreactive for VIP/nNOS/HU amounted to 7 and 4%, respectively. GAL/VIP-co-immunoreactivity was demonstrated in 69 and 27% of spiny neurons, respectively. We conclude that the number of neurons displaying co-reactivity for VIP and nNOS is a quantitative indicator of spiny neurons in both small and large intestine and that the proportion of spiny neurons is about 7% in small and 4% in large intestines. Since nerve fibres co-staining for NF/VIP/nNOS were found mainly in the circular muscle layer but not the surrounding perikarya of spiny neurons, we suggest that they may represent inhibitory motor neurons rather than descending interneurons.

Erratum: COX-2, TFF1, and Src define better prognosis in young patients with gastric cancer (Journal of Surgical Oncology (2013) 108:6 DOI: 10.1002/jso.23416))

In the original article, “COX‐2, TFF1, and Src define better prognosis in young patients with gastric cancer,” by Schildberg C, Abbas M, Merkel S, Agaimy A, Dimmler A, Schlabrakowski A, Croner R, Leupolt J, Hohenberger W, Allgayer H, published in the November 2013 issue of the Journal of Surgical Oncology (J Surg Oncol. 2013 Nov; 108:6; doi:10.1002/jso.23416), the author list contained errors. The names should instead read Schildberg CW, Abba M, Merkel S, Agaimy A, Dimmler A, Schlabrakowski A, Croner R, Leupolt JH, Hohenberger W, Allgayer H. We apologize for the error.