Unlike biochemical tests, DNA analysis permits the unambiguous identification of MEN2A gene carriers.
The mAb HCA2 reacts preferentially with HLA-A locus heavy chains. Its reactivity contrasts with that of HC10, a mAb with more pronounced specificity for HLA-B and -C heavy chains. Both HCA2 and HC10 were raised against free class I heavy chains of HLA-A and -B antigens respectively, to obtain mAbs that would still react with denatured class I antigens, as they occur in Western blotting, conventional light microscopical analysis of formalin-fixed, paraffin-embedded sections, and cryo-immuno-electron microscopy. HCA2 and HC10 indeed retain strong reactivity with free class I heavy chains in Western blots. The HCA2 mAb in particular reacts in a locus-specific manner by biochemical criteria. Conditions are described for use of HCA2 and HC10 in immunohistochemical staining of formalin-fixed, paraffin-embedded sections. HCA2 and HC10 also produce strong reactivity in immuno-electron microscopy. Their use allows the determination of tissue and subcellular distribution of class I antigens.
In human breast cancer, c‐Src activity is elevated compared to normal breast tissue. It is not yet known whether this increase in c‐Src activity is accompanied by an increase in c‐Src protein expression. In this study, c‐Src activity and protein expression were determined in a series of human breast cancers and in normal breast tissue, using immune complex kinase assays and immunoblotting. As the heterogeneity of breast cancer is not taken into account in these biochemical experiments, immunohistochemistry was also used to distinguish between normal and malignant cells. In human breast cancers, the c‐Src activity is increased 4‐ to 30‐fold, compared with normal breast tissue. This enhanced activity is accompanied by an increase in c‐Src protein expression, as shown by both immunoblotting and immunohistochemistry. Immunohistochemistry indicates that the majority of c‐Src appears to be concentrated around the nucleus in malignant cells, whereas in normal cells, it is distributed more evenly in the cytoplasm. These data confirm that c‐Src activity is increased in human breast cancer. In addition, this study provides strong immunohistochemical evidence that the c‐Src protein is also overexpressed, enabling a distinction to be made between normal and malignant cells.
Overexpression of EGFR and c-erbB2 frequently occurs in human breast cancers, correlating with poor prognosis. Here we show that overexpression of EGFR and c-erbB2 in cell lines increases cell migration, an important step in metastasis formation. The effect of EGFR on migration is dependent on the addition of EGF to the cells. In contrast, c-erbB2 seems to act independently of its ligand in these assays. Overexpression of this receptor is sufficient to induce cell migration. In addition, we investigated the involvement of a number of signal transduction pathways known to be activated by the EGFR. We found that inactivation of MAPKK results in a decreased migration, while inactivation of PI3K increases migration.z 1998 Federation of European Biochemical Societies.
We reassessed renal biopsy specimens from 116 patients with systemic lupus erythematosus and clinical manifestations of lupus nephritis to determine the contributions of the World Health Organization classification system, the activity and chronicity indexes of the National Institutes of Health scoring system, and various clinical parameters at the time of biopsy to predicting disease outcome. Multivariate analysis showed that only a chronicity index >3 was predictive for decreased renal survival, while age >31 years at biopsy and a chronicity index >3 were associated with decreased patient survival. Clinical tests of renal function were not reliable in discriminating between active lesions and chronic renal damage.Despite recent impressive improvements in patient survival compared with the presteroid era, renal disease secondary to systemic lupus erythematosus (SLE) is still associated with high morbidity and mortality rates (14). At present, uncertainty still exists about the prognostic value of various parameters in lupus nephritis, but recent cumulative evidence indicates that the presence of the following parameters at biopsy is associated with increased risk for renal failure: young age (<23 years), increased serum creatinine levels, diffuse proliferative lesions (World Health Organization [WHO] class IV), and a high chronicity index on renal histologic analysis (4-10).In this study, we analyzed 116 SLE patients with clinical evidence of lupus nephritis to determine the contribution of various factors to their prognosis and, in particular, the merits of evaluating renal histology in these patients. We also examined possible correlations between clinical and histologic data at the time of renal biopsy to see if the biopsy findings could have been predicted by simple clinical data determined at the time of biopsy.
PATIENTS AND METHODSPatients. In order to obtain an adequate study population, all 8 university hospitals and major teaching hospitals treating patients with SLE in The Netherlands were asked to cooperate in this study. Nearly 20% of those contacted declined cooperation for various reasons (disagreement with study protocol, insufficient time). The remaining 80% of clinicians agreed to provide data and renal biopsy specimens from all their patients with lupus nephritis who had undergone biopsy. We obtained 152 renal biopsies, 10 of which were repeat biopsies and were therefore not included in our analysis; another 26 samples had to be excluded for various reasons (14 due to insufficient material [less than 6 glomeruli], 9 because the diagnosis of SLE did not meet the American Rheumatism Association [ A M ] criteria [ 11.121, and 3 because postbiopsy followup, before an end-point of either death or renal dialysis was reached. was
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