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Sökning: WFRF:(Wester Kenneth)

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71.
  • Wester, Kenneth, et al. (författare)
  • HER-2 : A possible target for therapy of metastatic urinary bladder carcinoma
  • 2002
  • Ingår i: Acta Oncologica. - : Informa UK Limited. - 0284-186X .- 1651-226X. ; 41:3, s. 282-288
  • Tidskriftsartikel (refereegranskat)abstract
    • Human epidermal growth factor receptor 2, HER-2, is overexpressed in various tumours, e.g. breast- and bladder tumours. The aim of this study was to predict the potential use of HER-2 receptors as targets in systemic treatment of disseminated bladder tumours. HER-2 expression was assessed in bladder carcinoma metastases and the corresponding primary tumours, and subsequently compared with the EGFR expression. HER-2 and EGFR expression was analysed by immunohistochemistry in formalin-fixed, paraffin-embedded tissues from 21 patients with metastatic bladder carcinoma. HER-2 was overexpressed in 81% of the primary tumours and in 67% of the metastases. All HER-2-positive metastases were from HER-2-positive primary tumours. The results for EG FR were 71% of both primary and metastases-positive tumours. In 90% of the primary tumours and 86% of the metastases, at least one of the receptors was overexpressed. These results suggest that HER-2 targeted therapy can be considered as an alternative or a complement to other modalities in the treatment of metastatic urinary bladder carcinoma.
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72.
  • Wester, Kenneth (författare)
  • Quantitative immunochemistry in tissue sections : With special reference to urinary bladder carcinoma
  • 1999
  • Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
    • Urinary bladder carcinoma affects about 2 000 people in Sweden every year. It is a heterogeneous disease and for adequate treatment decisions, prognostic tools are needed. Many of these are based on immunohistochemistry (MC) and hampered by poorly reproducible methodologies and subjective evaluations.This investigation aimed to evaluate and define the conditions for an objective and reproducible quantification of(MC) applied to routinely processed (formalin-fixed and paraffin-embedded) tissue material. To establish theseconditions, proliferation-related proteins, tumour-suppressor proteins and endothelial cell markers were studiedThree computerised colour-based image analysis methods, aiming to reduce user interaction, were developed and evaluated for variability and reproducibilty in quantification of IHC applied to histological sections. Themethods proved stable to variations in focus- and light-settings but revealed a sensitivity to poor IHC-qualityand choice of chromogen. Implementation of an external control system, used as base for computing theclassifier, further increased the stability.Using these methods, effects of varying tissue-handling and immunostaining conditions were analysed. Storageof both paraffin-blocks and -sections was deleterious to immunoreactivity but could be compensated for bytayloring of the heat induced epitope retrieval (HIER) protocol.A control system for MC based on cultured cells was developed to reflect variations in tissue-handling and IHC-conditions. Control and biopsy material showed similar sensitivity to section storage, MC- and HIER-protocolvariations. Furthermore, controls reflected variations in fixation time and proved stable to variations in sectionthickness.
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73.
  • Wester, Kenneth, et al. (författare)
  • Zinc-based fixative improves preservation of genomic DNA and proteins in histoprocessing of human tissues.
  • 2003
  • Ingår i: Lab Invest. - 0023-6837. ; 83:6, s. 889-99
  • Tidskriftsartikel (refereegranskat)abstract
    • Advantageous preservation of histology and detailed cellular morphology has rendered neutral buffered formalin (NBF) the most widely used fixative in clinical pathology. Despite excellent morphology for routine diagnostics, a major drawback of NBF fixation is its detrimental effect on DNA and RNA quality. In addition to complicating analysis of genes and transcripts in complex tissues, NBF denatures proteins and thereby hampers immunohistochemical visualization of certain antigens. In the present study, we evaluated a zinc-based fixative (ZBF) regarding its effects on tissue morphology, quality of genomic DNA, and preservation of protein immunoreactivity in a broad spectrum of tissues. Four different modes of fixation were analyzed: ZBF-paraffin embedding, NBF-paraffin embedding, ZBF-fixation prior to snap-freezing, and immediate snap-freezing. Laser-assisted microdissection, allowing retrieval of a defined number of cells for PCR, was used to study DNA quality. Genomic DNA was analyzed using primers for beta2-microglobulin and the transferrin receptor. Immunohistochemistry was performed using nine antibodies. Tissue microarray blocks were used for analysis of morphology and immunoreactivity. Only slight impairment of morphologic qualities was found after ZBF-paraffin embedding, whereas ZBF prior to freezing resulted in a more crisp morphology compared with routine cryosections. A significantly higher DNA yield was observed in samples isolated from ZBF-paraffin-embedded tissues compared with NBF-paraffin-embedded tissues. Both yield and quality of DNA was comparable in frozen tissues irrespective to prior ZBF fixation. Immunoreactivity in paraffin-embedded tissue was superior in ZBF-fixated tissue compared with NBF-fixated for a majority of tested antibodies. Furthermore, for seven out of nine antibodies, antigen retrieval pretreatment proved unnecessary in ZBF-fixated tissue. Thus, despite a slight impairment of morphology, ZBF preserves protein structures well. We conclude that ZBF is superior to NBF for analysis of DNA and protein expression. Fixation of tissues in ZBF may also be an alternative strategy to freeze storage of tissue specimens, eg, in future bio-banks.
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74.
  • Älgenäs, Cajsa, et al. (författare)
  • Antibody performance in western blot applications is context- dependent
  • 2014
  • Ingår i: Biotechnology Journal. - : Wiley. - 1860-6768 .- 1860-7314. ; 9:3, s. 435-445
  • Tidskriftsartikel (refereegranskat)abstract
    • An important concern for the use of antibodies in various applications, such as western blot (WB) or immunohistochemistry (IHC), is specificity. This calls for systematic validations using well-designed conditions. Here, we have analyzed 13000 antibodies using western blot with lysates from human cell lines, tissues, and plasma. Standardized stratification showed that 45% of the antibodies yielded supportive staining, and the rest either no staining (12%) or protein bands of wrong size (43%). A comparative study of WB and IHC showed that the performance of antibodies is application-specific, although a correlation between no WB staining and weak IHC staining could be seen. To investigate the influence of protein abundance on the apparent specificity of the antibody, new WB analyses were performed for 1369 genes that gave unsupportive WBs in the initial screening using cell lysates with overexpressed full-length proteins. Then, more than 82% of the antibodies yielded a specific band corresponding to the full-length protein. Hence, the vast majority of the antibodies (90%) used in this study specifically recognize the target protein when present at sufficiently high levels. This demonstrates the context- and application-dependence of antibody validation and emphasizes that caution is needed when annotating binding reagents as specific or cross-reactive. WB is one of the most commonly used methods for validation of antibodies. Our data implicate that solely using one platform for antibody validation might give misleading information and therefore at least one additional method should be used to verify the achieved data.
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