摘要:

AbstractWe have investigated the use of a novel technique,in situend‐labelling, as a means of the specific identification of apoptotic cells in formalin‐fixed, paraffin‐processed tissue sections. The technique relies on the presence of DNA strand breaks in apoptotic cells, caused by activation of endogenous nuclease activity during the process of cell death. These strands are labelled with a non‐isotopic reporter molecule in the presence of a DNA polymerase, and labelled DNA is identified immunohistochemically. We show thatin situend‐labelling stains cells with the morphological characteristics of apoptosis, and greatly simplifies their identification. Furthermore, in two model systems, the number of labelled cells parallels the number of cells undergoing apoptosis as measured by alternative techniques. The ability of the Klenow fragment of DNA polymerase to label apoptotic nuclei suggests that the characteristic DNA fragmentation seen during this process involves the formation of DNA breaks with a 5′ overhang.in situend‐labelling will be valuable for the identification and quantitation of apoptosis in a range of normal tissues and in a variety of pathological states. However, the technique is not specific for programmed cell death, and results must be interpreted with caution and correlated with morphological criteria

ISSN:0022-3417    

DOI:10.1002/path.1711700102

出版商:John Wiley&Sons, Ltd.    

年代:1993

数据来源: WILEY

摘要:

AbstractMELAS (mitochondrial myopathy, encephalopathy, lactic acidosis, and stroke‐like episodes) is a clinically devastating disease of children and young adults. The cause of the stroke‐like episodes is not known. We have sequenced the mitochondrial DNA (mtDNA) in archival paraffin‐embedded material from two cases. In only one of these did the mitochondrial tRNALeu(UUR) gene contain the nucleotide 3243 A‐to‐G mutation that is most commonly responsible for MELAS. In this case, we determined the relative proportion of mutant:wild‐type mtDNA in sections of the central nervous system and other tissues by PCR amplification,PalI digestion, DNA electrophoresis, and scanning densitometry of the ethidium bromide‐stained gels. The technique allowed the proportion of mitochondria that contain the mutant genome to be compared with the histological findings in immediately adjacent sections of tissue. The mutant mtDNA was detectable in most tissues, the percentage of mtDNA ranging from barely detectable levels to 78 percent. The relative amount of mutant mtDNA correlated poorly with the distribution of histological lesions, both within the central nervous system and in other tissues examined. The proportion was high in tissues such as liver, kidney, adrenal, and pancreas that appeared histologically normal. Relatively low levels were present in some regions of the central nervous system, such as the occipital lobe, which contained many of the characteristic infarct‐like lesions. These observations do not support previous speculation that the distribution of these lesions reflects that of the defective mitochondria. The results emphasize the usefulness of the polymerase chain reaction in correlative histo

ISSN:0022-3417    

DOI:10.1002/path.1711700103

出版商:John Wiley&Sons, Ltd.    

年代:1993

数据来源: WILEY

摘要:

AbstractTGF alpha and beta expression was examined using rabbit polyclonal antibodies and immunohistochemistry on a series of 195 breast carcinomas. TGF alpha immunoreactivity was observed in all but nine of the tumours, with over 50 per cent staining strongly. The polyclonal TGF alpha antibody (CIM1), when compared with a commercially available mouse monoclonal TGF alpha antibody used on the same sections, gave a good correlation (r=0.52,P<0.001). Both TGF alpha antibodies produced a granular cytoplasmic staining pattern, that with CIM 1 being coarser, suggestive of binding to an aggregated protein or organelle. Eighty‐one per cent of tumours stained with the TGF beta antibody, 35 per cent strongly. There was significant co‐expression of TGF alpha and TGF beta (P<0.001). However, they were not found to be useful prognostic indicators, lacking any significant correlation with histological classification, tumour size, nodal status, oestrogen receptor status, S‐phase fraction, or overall survival over a 9–12 year period. The expression of these growth factors in most breast carcinomas suggests that they have important biological roles, but the exact nature of these roles remains unclear at the

ISSN:0022-3417    

DOI:10.1002/path.1711700104

出版商:John Wiley&Sons, Ltd.    

年代:1993

数据来源: WILEY

摘要:

AbstractThe effect of the differentiating agent all‐trans‐retinoic acid on the expression of components of the milk fat globule membrane and HLA‐DR by breast cancer cell lines has been examined. Effects on proliferation were also considered, to determine whether any cell surface changes were related to or independent of proliferation effects. No significant differences were observed in the expression of components detected by the milk fat globule membrane antibodies HMFG1 and HMFG2 over 8 days culture with 10‐7‐10‐9M retinoic acid for the cell lines MCF‐7, T‐47 D, ZR‐75, MDA‐MB‐231, and BT‐20. In contrast, there was enhanced expression of HLA‐DR by two oestrogen receptor‐positive cell lines T‐47 D and ZR‐75 and the oestrogen receptor‐negative line MDA‐MB‐231, with differing sensitivities. These effects were independent of inhibition of proliferation, which was only observed for oestrogen receptor‐positive cell lines and for different durations of exposure. The finding of enhanced HLA‐DR expression after retinoic acid treatment has not previously been reported and is of interest regarding clinical poten

ISSN:0022-3417    

DOI:10.1002/path.1711700105

出版商:John Wiley&Sons, Ltd.    

年代:1993

数据来源: WILEY

摘要:

AbstractAndrogen actions and androgen receptors (ARs) have been described in human breast cancer cells bothin vivoandin vitro. With the use of a new monoclonal anti‐AR antibody, AR was immunohistochemically demonstrated in 76 primary breast cancers. Positive immunostaining was found in 79 per cent of tumours. Benign ductal epithelium was often AR‐positive whereas the tumour stroma lacked AR immunoreactivity. At the subcellular level, nuclear localization was evident using either cross‐linking (Zamboni's fluid) or precipitating (acetone) fixatives on frozen sections. The use of archival paraffin‐embedded tissue yielded negative results. A significant association was found between expression of AR and oestrogen receptor (ER) (P=0.0006) determined immunohistochemically on adjacent sections. Most progesterone receptor (PR)‐negative cases were also AR‐negative (P=0.02), but significant proportion (38 per cent) of AR‐positive tumours did not contain PR. Unlike ER, AR was not associated with aneuploidy orerb‐B2 oncogene overexpression, and was only marginally associated with tumour proliferation rate (S‐phase fraction by DNA flow cytometry). In conclusion, the close association of AR with ER and PR suggests that immunohistochemical determination of androgen receptors may have value as a prognostic factor and/or predictor of response to

ISSN:0022-3417    

DOI:10.1002/path.1711700106

出版商:John Wiley&Sons, Ltd.    

年代:1993

数据来源: WILEY

摘要:

AbstractThis study was aimed at assessing the effects of therapeutic long‐term administration of androgens on normal human female breast. Sections from mastectomy specimens of 29 female‐to‐male transsexuals who had prolonged androgen administration prior to surgery were examined using routine light microscopy and immunohistochemical techniques. For comparison, sections from ten ‘normal’ female breast reduction mammoplasty specimens and ten cases of gynaeco‐mastia were similarly examined. Haematoxylin and eosin‐stained sections were assessed for the prevalence of elements of the normal breast and benign breast lesions. Immunoperoxidase techniques were performed to study the distribution of a variety of breast‐associated antigens and receptors. The results were assessed semi‐quantitatively.The prevalence of normal acini and ducts, fibrosis, cysts, and apocrine metaplasia in transsexual specimens was not statistically different from that seen in normal controls. However, transsexual specimens had a significantly higher prevalence of microcalcification than normals. The majority of transsexual specimens were positive for gross cystic disease fluid protein‐15, lactoferrin, and progesterone and oestrogen receptors, and negative for B72.3 and pS2. These findings were not significantly different from those in normal controls.All ten gynaecomastia specimens were positive for oestrogen and progesterone receptors. The prevalence of oestrogen receptors was significantly higher than that seen in transsexuals and normal controls, but the prevalence of progesterone receptors was only significantly higher than that seen in transsexuals.It is concluded that long‐term androgen administration does not appear to have any significant lasting effect on the normal human female breast, as demonstrated by a wide range of histological and immuno

ISSN:0022-3417    

DOI:10.1002/path.1711700107

出版商:John Wiley&Sons, Ltd.    

年代:1993

数据来源: WILEY

摘要:

AbstractThis is a combined study usingin situhybridization and the polymerase chain reaction to investigate the presence of Coxsackie virus RNA in formalin‐fixed tissue from cases of childhood myocarditis. Of the ten cases studied, two were positive by both methods. The virus RNA was predominantly located in areas showing an inflammatory cell infiltrate and myofibre necrosis. These findings suggest that direct lytic infection of myocytes by virus is responsible for myocaiditis in these cases, rather than an autoimmune process, which has been suggested previously. The findings in one case, where the virus showed a marked sub‐endocardial distribution, may have implications for the aetiology of endocardial fibroelastosis by confirming a viral tropism for this location. The techniques used in this study are easily repeatable and can be directly applied to look for viruses in a number of other diseases where a viral aetiology is suspec

ISSN:0022-3417    

DOI:10.1002/path.1711700108

出版商:John Wiley&Sons, Ltd.    

年代:1993

数据来源: WILEY

摘要:

AbstractWe have recently demonstrated that killer cells expressing a cytolytic factor, perforin, infiltrate the hearts of mice with acute viral myocarditis and may play an important role in the mechanism of myocardial damage. To clarify the mechanism ofin vivocardiac myocyte injury mediated by perforin, we investigated the release of perforin molecules from killer cells by immunoelectron microscopy and examined the circular lesions formed by perforin on the membrane of cardiac myocytes. We found that there was massive release of perforin molecules from the killer cells directly onto the surface of the cardiac myocytes. Furthermore, electron microscopy of ultrathin ventricular sections treated with trypsin revealed numerous circular lesions with the characteristics of perforin pores, in the membranes of cardiac myocytes. These findings provide the first direct evidence that killer cells injure cardiac myocytes by releasing perforin and may play a critical role in the myocardial damage occurring in acute viral myocarditis.

ISSN:0022-3417    

DOI:10.1002/path.1711700109

出版商:John Wiley&Sons, Ltd.    

年代:1993

数据来源: WILEY

摘要:

AbstractThe cellular distribution of heart‐type fatty acid‐binding protein (H‐FABP) immunoreactivity was examined in normal human tissues using a polyclonal antibody against human H‐FABP. Immunoreactivity was detected in cardio‐myocytes of both ventricles and atria as well as in all striated muscles investigated. In addition, staining was frequently observed in parietal cells of the stomach, renal epithelial cells, acinar and ductal cells of the breast, ductal cells of the salivary gland, corpus luteum, and Leydig cells of the testis. Adipocytes and vascular endothelial cells were positive but other tissues and cells examined were negative. Old infarcts of the heart replaced by fibrous connective tissues were not labelled. Necrotic cardiomyocytes and morphologically normal cardiomyocytes in acute ischaemic lesions 1 h after onset showed reduced or no H‐FABP immunoreactivity. Thus, decreased immunoreactivity for H‐FABP may be a good histological marker of damaged c

ISSN:0022-3417    

DOI:10.1002/path.1711700110

出版商:John Wiley&Sons, Ltd.    

年代:1993

数据来源: WILEY

摘要:

AbstractIn this study we investigated immunohistochemically the expression of p53 and c‐erbB‐2 proteins in 30 gall bladder adenocarcinomas, one carcinomain situ, eight gall bladder epithelial dysplasias, and four cases of chronic cholecystitis. p53 expression could be found in 14 (47 per cent) adenocarcinomas and in two out of eight epithelial dysplasias. There were significantly more p53‐positive grade II‐III tumours than grade I tumours (P=0.032 according to Fisher's exact probability test). c‐erbB‐2 expression was found in three (10 per cent) adenocarcinomas, but all dysplasias were c‐erbB‐2‐negative. All three c‐erbB‐2‐positive cases were also p53‐positive. The results indicate that p53 mutations and c‐erbB‐2 gene alterations play a role in the neoplastic transformation of gall bladder epithelial cells. Co‐expression of p53 and c‐erbB‐2 suggests that alterations of these genes might act in concert in the neoplastic transformation. The occurrence of p53 expression in gall bladder dysplasias suggests that p53 mutations could be an early event in the evolution of some gall bladder carcinomas, as has been suggested for some other types of tumours, su

ISSN:0022-3417    

DOI:10.1002/path.1711700111

出版商:John Wiley&Sons, Ltd.    

年代:1993

数据来源: WILEY