Tissue microarrays from HOPE-fixed specimens allow for enhanced high throughput molecular analyses in paraffin-embedded material

Goldmann, Torsten; Drömann, Daniel; Marzouki, M.-N.; Schimmel, Udo; Debel, Karsten; Branscheid, D.; Zeiser, Tobias; Rupp, Jan; Gerdes, Johannes; Zabel, P.; Vollmer, Ekkehard

doi:10.1016/j.prp.2005.07.006

Cited by 18 publications

(11 citation statements)

References 11 publications

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“…2005). The thickness of the cell wall of mycobacteria increases its resistance to simple disruption procedures and hinders the release of bacterial nucleic acids (Goldmann, Dromann, Marzouki, Schimmel, Debel, Branscheid, Zeiser, Rupp, Gerdes, Zabel & Vollmer 2005). There is also evidence that molecules which inhibit PCR may be introduced during tissue processing (Bascuñana & Belák 1996).…”

Section: Discussionmentioning

confidence: 99%

Detection and identification of aquatic mycobacteria in formalin‐fixed, paraffin‐embedded fish tissues

Pourahmad

Thompson

Adams

et al. 2009

Journal of Fish Diseases

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The isolation of mycobacteria from field samples is problematic, and isolation of the bacterium is sometimes not even attempted. The detection of mycobacteria through traditional histology using formalin-fixed, paraffin-embedded (FFPE) tissues is neither sensitive nor specific. However, detection of mycobacterial DNA from FFPE specimens, suspected of being infected with mammalian mycobacteriosis, is a routine clinical procedure. In the present study, a polymerase chain reaction (PCR)-based method was used to detect and identify mycobacteria in FFPE specimens sampled from fish suspected of being infected with fish mycobacteriosis. A total of 45 fish tissue samples, comprising of 12 tissue samples obtained from experimentally infected fish and the remainder from fish naturally infected with mycobacteria, were analysed using a PCR protocol which amplifies a fragment of the mycobacterial 65 kDa heat-shock protein (hsp65) gene. PCR-restriction enzyme analysis and/or sequencing were employed to further analyse the PCR amplicons. The PCR results were compared with those obtained by histology and culture. Mycobacterial DNA was detected in 34 of the 45 samples examined, of which 16 samples (47%) showed granulomatous reactions on histological examination. Using histology as the gold standard, no false-negative PCR results were obtained. Also, considering the presence or absence of granulomas as a diagnostic criterion, the sensitivity and specificity of PCR in 42 of the FFPE tissues were 16/16 (100%) and 8/26 (approximately 30.8%), respectively. Corresponding microbiological cultures were available for 15 cases, of which 13 were pure Mycobacterium cultures. Of these, 13 were PCR positive (100% sensitivity and 50% specificity). The PCR-based methods used here proved sensitive, specific and rapid for the detection of mycobacteria in routinely processed paraffin wax-embedded and formalin-fixed histological samples, and the results of the study suggest that this method has potential use in retrospective epidemiological studies.

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Section: Discussionmentioning

confidence: 99%

Detection and identification of aquatic mycobacteria in formalin‐fixed, paraffin‐embedded fish tissues

Pourahmad

Thompson

Adams

et al. 2009

Journal of Fish Diseases

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“…Tissue microarrays from HOPE-fixed, paraffin-embedded tissues were produced as described by Goldmann and colleagues (16). HOPE-fixed cells were dehydrated and subsequently paraffin-embedded according to Marwitz and colleagues (17) to produce cell blocks for immunocytochemistry.…”

Section: Production Of Tissue Microarray and Cellblocksmentioning

confidence: 99%

Downregulation of the TGFβ Pseudoreceptor BAMBI in Non–Small Cell Lung Cancer Enhances TGFβ Signaling and Invasion

et al. 2016

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Non-small cell lung cancer (NSCLC) is characterized by early metastasis and has the highest mortality rate among all solid tumors, with the majority of patients diagnosed at an advanced stage where curative therapeutic options are lacking. In this study, we identify a targetable mechanism involving TGFb elevation that orchestrates tumor progression in this disease. Substantial activation of this pathway was detected in human lung cancer tissues with concomitant downregulation of BAMBI, a negative regulator of the TGFb signaling pathway. Alterations of epithelialto-mesenchymal transition (EMT) marker expression were observed in lung cancer samples compared with tumor-free tissues. Distinct alterations in the DNA methylation of the gene regions encoding TGFb pathway components were detected in NSCLC samples compared with tumor-free lung tissues. In particular, epigenetic silencing of BAMBI was identified as a hallmark of NSCLC. Reconstitution of BAMBI expression in NSCLC cells resulted in a marked reduction of TGFb-induced EMT, migration, and invasion in vitro, along with reduced tumor burden and tumor growth in vivo. In conclusion, our results demonstrate how BAMBI downregulation drives the invasiveness of NSCLC, highlighting TGFb signaling as a candidate therapeutic target in this setting. Cancer Res; 76(13); 3785-801. Ó2016 AACR.

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“…After generation of numerous hybridomas, the corresponding monoclonal antibodies were subjected to primary screening using cell cultures and human tissues. One of the clones directed against a cytoplasmic fraction of AECII showed reactivity with AECII, alveolar macrophages and adenocarcinomas of the lungs, which was further verified using tissue microarrays (TMAs) from NSCLC tissues treated with the HOPE technique (Srinivasan et al , 2002; Goldmann et al , 2003, 2005). This clone was designated MAdL (marker for adenocarcinomas of the lung).…”

mentioning

confidence: 73%

“…For a preliminary study of the immune reactivity of MAdL, TMAs were produced from 35 HOPE-fixed, paraffin-embedded NSCLC tissues as previously described (Goldmann et al , 2005). HOPE-fixed specimens allow immunohistochemistry (IHC) without antigen retrieval (AR), which could create artefacts in staining or morphology (Goldmann et al , 2003).…”

Section: Methodsmentioning

confidence: 99%

Generation and evaluation of a monoclonal antibody, designated MAdL, as a new specific marker for adenocarcinomas of the lung

et al. 2011

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Background:Different therapy regimens in non-small-cell lung cancer (NSCLC) are of rising clinical importance, and therefore a clear-cut subdifferentiation is mandatory. The common immunohistochemical markers available today are well applicable for subdifferentiation, but a fraction of indistinct cases still remains, demanding upgrades of the panel by new markers.Methods:We report here the generation and evaluation of a new monoclonal antibody carrying the MAdL designation, which was raised against primary isolated human alveolar epithelial cells type 2.Results:Upon screening, one clone (MAdL) was identified as a marker for alveolar epithelial cell type II, alveolar macrophages and adenocarcinomas of the lung. In a large-scale study, this antibody, with an optimised staining procedure for formalin-fixed tissues, was then evaluated together with the established markers thyroid transcription factor-1, surfactant protein-A, pro-surfactant protein-B and napsin A in a series of 362 lung cancer specimens. The MAdL displays a high specificity (>99%) for adenocarcinomas of the lung, together with a sensitivity of 76.5%, and is capable of delivering independent additional diagnostic information to the established markers.Conclusion:We conclude that MAdL is a new specific marker for adenocarcinomas of the lung, which helps to clarify subdifferentiation in a considerable portion of NSCLCs.

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Tissue microarrays from HOPE-fixed specimens allow for enhanced high throughput molecular analyses in paraffin-embedded material

Cited by 18 publications

References 11 publications

Detection and identification of aquatic mycobacteria in formalin‐fixed, paraffin‐embedded fish tissues

Detection and identification of aquatic mycobacteria in formalin‐fixed, paraffin‐embedded fish tissues

Downregulation of the TGFβ Pseudoreceptor BAMBI in Non–Small Cell Lung Cancer Enhances TGFβ Signaling and Invasion

Generation and evaluation of a monoclonal antibody, designated MAdL, as a new specific marker for adenocarcinomas of the lung

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