Low prevalence of Epstein-Barr virus in incident gastric adenocarcinomas from the United Kingdom.

Epstein-Barr virus has been associated with a proportion of typical gastric adenocarcinomas. Here we report that the prevalence of Epstein-Barr virus in gastric adenocarcinomas from the United Kingdom is one of the lowest in the World. Gastric adenocarcinoma is another tumour whose association with Epstein-Barr virus varies with the population studied.

The long term results of endoscopic surveillance of premalignant gastric lesions.

BACKGROUND: A large proportion of patients attending open access endoscopy have histological and gross pathological findings that are potentially premalignant. The proportion of these patients who go on to develop malignancies and the timescale over which this occurs are uncertain. AIMS: This study aims to discover the incidence of gastric cancers in this "high risk" group and to examine the potential for their early diagnosis and treatment. PATIENTS: A total of 1753 patients attended open access endoscopy. From these, 166 patients with dysplasia, intestinal metaplasia, atrophic gastritis, foveolar hyperplasia, regenerative changes, polyps, or ulcers who agreed to undergo annual surveillance endoscopy were studied. METHODS: Patients were endoscoped annually. Additionally, patients with ulcers were re-examined at two monthly intervals until ulcer healing. Cancers detected were treated by gastrectomy. RESULTS: Twenty two of 1753 patients attending open access endoscopy had gastric cancer (1.3%). In the study population, 14 cancers were detected over 10 years (8.4 %). These were of an earlier stage than those detected at open access (stage I and II 67% v 23%; p<0.05) and five year survival was significantly higher (50% v 10%; p=0.006). In atrophic gastritis and intestinal metaplasia the risk of malignancy was 11%. CONCLUSIONS: In patients with atrophic gastritis or intestinal metaplasia, annual surveillance can detect most new tumours at an early stage with a major improvement in survival. Potential benefits of such a surveillance programme are large and warrant further investigation in a multicentre randomised controlled trial.

The diagnosis of invasive lobular breast carcinoma. Does MRI have a role?

Invasive lobular breast carcinoma accounts for approximately 15% of all breast cancers and is difficult to detect using conventional breast imaging techniques. We report a comparison between clinical, ultrasound scan (USS), mammographic and magnetic resonance imaging (MRI) of 22 patients with invasive lobular breast carcinomas. Actual tumour size was ascertained by histopathology. MRI detected 21 of the 22 invasive lobular cancers whilst mammography and USS detected 16 and 20 respectively. 19 tumours were clinically palpable. MRI was more accurate at assessing tumour size than USS and clinical examination, both of which underestimated tumour size.

Spontaneous regression of metastatic malignant melanoma of the parotid gland and neck lymph nodes: a case report and a review of the literature.

We report the case history of a patient with complete spontaneous regression of metastatic cutaneous melanoma with parotid and neck lymph node metastases. Complete spontaneous regression of metastatic melanoma is very rare, with an estimated incidence of 0.22%-0.27%. We review the literature on this subject. Elucidation of the process of spontaneous regression may offer the possibility of improved methods of treating and preventing cancer.

Schwannoma (neurilemmoma) of the tonsil.

Schwannoma of the tonsil is an extremely uncommon clinical entity with only one reported case in an adult in the medical literature to date. We report, to our knowledge, the first known case in a child.

Redefining surgery for gastric cancer.

BACKGROUND: Despite encouraging retrospective and non-randomized trials, two large prospective, randomized trials of D1 vs D2 resections show double the mortality in the D2 group, with no increase in long-term survival. However, the D2 resection still offers the only hope of cure when N2 nodes are involved. We propose a reclassification of the International Union Against Cancer TNM "N" staging to a system with an anatomical basis that is useful in defining the surgery performed. Junctional nodes lying between the N1 and N2 tiers will act as a guide to surgery. Where these nodes are uninvolved, the probability of gastric bed (N2) involvement is low and the radical D2 dissection with its higher mortality and morbidity can be avoided.CONCLUSION: Such "stage-appropriate" surgery will reduce the number of D2 resections while ensuring that patients with N2 disease are not denied curative surgery. A prospective, randomized, controlled trial of targeted surgery is required.

Rapid peroperative assessment of axillary lymph node status using imprint cytology.

Sentinel lymph node biopsy for invasive breast cancer is a new technique that has been shown to be accurate in staging the axilla. Patients with a positive sentinel node will potentially need a second operation to clear the axilla. A reliable technique for assessing lymph nodes intraoperatively could potentially avoid this second procedure. This study aimed to assess the accuracy of peroperative imprint cytology of axillary lymph nodes in patients with invasive breast carcinoma. A total of 157 nodes were studied. One-hundred and nineteen nodes were both negative on imprint cytology and paraffin section. Thirty-eight nodes were positive on imprint cytology. Of these, 37 were positive on histology. Imprint cytology is a rapid technique that is inexpensive and has been shown to be reliable in this and other studies. It may prove to be of value in patients undergoing sentinel lymph-node biopsy for breast carcinoma.

Simple method for pretreatment of tissue sections for the detection of apoptosis by in situ end-labelling and in situ nick translation.

Aims-To overcome the problems associated with proteolytic pretreatment of tissue sections for the detection of apoptosis.Methods-Formalin fixed, paraffin wax embedded tissue sections of reactive lymph nodes and biopsy specimens of Burkitt lymphoma were pretreated by pressure cooking for the detection of apoptosis using the in situ end-labelling and in situ nick translation methods.Results-The results achieved with the in situ end-labelling and nick translations methods were compared with those obtained using a novel anti-apoptosis specific protein (ASP) antibody. The staining patterns generated using the three methods were similar and consistent, although the ASP antibody seemed to be more sensitive and detected higher numbers of apoptotic cells within sections.Conclusions-Pressure cooking is advocated as an alternative method to proteolytic enzyme digestion for pretreating paraffin wax sections. It is reliable, inexpensive, reduces the need to optimise pretreatment variables for different tissues, and permits double immunostaining of sections.

Detection of the multidrug resistance marker P-glycoprotein by immunohistochemistry in malignant lung tumours.

BACKGROUND: The multidrug resistance marker P-glycoprotein (P-gp) was studied immunohistochemically in 78 primary malignant lung tumours. P-gp is a 170 kD transmembrane ATP dependent drug efflux pump which has been shown to be important in the resistance of some tumours to chemotherapy. Certain normal tissues express P-gp and tumours derived from these tissues are often insensitive to cytotoxic agents, showing raised P-gp levels innately or following chemotherapy or radiotherapy. METHODS: Samples from 78 patients undergoing surgery for primary malignant lung tumours were snap frozen and stained immunohistochemically using the monoclonal antibody C219 which reacts with a P-gp epitope. None of the study group had received chemotherapy or radiotherapy before surgery was performed. RESULTS: Twenty seven of the 78 lung tumours (34.6%) showed immunohistochemically detectable levels of P-gp which varied with tumour type; 17 of 54 squamous cell carcinomas (31.5%), seven of 15 adenocarcinomas (46.7%), and neither of two small cell carcinomas showing positive staining. In six of seven cases normal respiratory epithelium present showed the presence of P-gp. CONCLUSIONS: P-gp is immunohistochemically detectable in frozen tissue from a proportion of malignant lung tumours before exposure to radiotherapy or drugs associated with multidrug resistance. It may have a role in tumour resistance to cytotoxic drugs, but further clinical studies will be required to evaluate any correlation between P-gp levels and response to treatment.

Identification of the antigen recognized by the monoclonal antibody BU31 as lamins A and C.

The murine monoclonal antibody BU31 binds to the nuclear membrane of many cell types. The expression of the BU31 antigen has previously been shown to have an inverse correlation with the proliferative index in lung tumours, defined by Ki67 staining. The distribution of BU31-positive cells is now shown to parallel the distribution of non-dividing cells in a range of normal human and rat tissues, although neuroendocrine cells and germ cells in the testis show no reactivity. Cells grown in culture and induced to undergo growth arrest show a higher level of labelling with BU31 than their proliferating counterparts. Confocal laser scanning microscopy reveals that the BU31 antigen is distributed predominantly along the nuclear lamina, with occasional internal foci. This distribution is very similar to that of the nuclear membrane proteins lamin A and lamin C, suggesting that the BU31 antigen and lamins A and C could be one and the same. Immunoblotting using recombinant lamin proteins confirmed this proposal. Moreover, a monoclonal antibody to the non-proliferation-associated antigen, statin, also recognizes lamins A and C. These data indicate that the demonstration of lamins A and C can be used to provide information on the proliferative activity of normal and neoplastic tissues. These data also suggest a role for nuclear lamins A and C during cellular quiescence, possibly through the reorganization and maintenance of nuclear structure, or more directly through interactions with the retinoblastoma gene product or related proteins.

Detection of Ki67 antigen by a new sheep polyclonal antiserum.

This report describes the characterisation of a polyclonal sheep antiserum against the Ki67 antigen. On western blots, this antiserum recognises a pair of bands of high molecular weight identical with those seen with another polyclonal Ki67 antiserum and the MIB 1 monoclonal antibody. The new antiserum showed nuclear staining of a proportion of cells in paraffin wax embedded tissue sections following antigen retrieval using a microwave oven or pressure cooker. This staining pattern was blocked by incubating the serum with the peptide used as immunogen. The proportion and distribution of immunostained nuclei was identical with that seen with the alternative reagents that recognise the Ki67 antigen. The new reagent stained the same proportion of cells when used over a wide range of dilutions. There was no cross-reactivity with unrelated antigens sometimes detected by the monoclonal antibodies.

Measurement of proliferation in renal biopsy specimens: evidence of subclinical tubular damage in the nephrotic syndrome.

BACKGROUND: Proliferating cells can be detected in histological material in various ways. This investigation was to study the feasibility and usefulness of application of proliferation markers to routine renal biopsy specimens. METHODS: One hundred and thirteen renal biopsies fixed in formalin and embedded in paraffin were studied immunohistologically using antibody MIB 1, which recognises the Ki-67 antigen. Twenty-two biopsies were also immunostained with antibody PC10 against proliferating cell nuclear antigen. RESULTS: PC10 stained nuclei in all biopsies, including those that were negative with MIB 1. MIB 1 stained nuclei in endocapillary sites to various extents, and there was an average of less than one stained endocapillary nucleus per glomerulus in biopsies with IgA nephropathy and IgM nephropathy, conventionally regarded as types of proliferative glomerulonephritis. Glomerular extracapillary nuclei were stained by MIB 1 in vasculitic disorders and at the site of tip changes. MIB 1 also stained nuclei in the arterial intima, especially in vascular rejection, and in interstitial tissues, correlating with renal excretory function. Tubular nuclei stained by MIB 1 were common in biopsies from patients with renal impairment and in a group with the nephrotic syndrome, in many of whom renal function was normal. CONCLUSIONS: The main conclusions are that (1) immunohistological markers of proliferation can be applied to routine renal biopsy material; (2) PC10 appears to overestimate proliferation compared with MIB 1; and (3) there is evidence of subclinical tubular damage in the nephrotic syndrome, shown by increased tubular proliferation without clinical renal impairment. This observation seems not to have been made previously.

Down-regulation but not phosphorylation of stathmin is associated with induction of HL60 cell growth arrest and differentiation by physiological agents.

Stathmin is a cytosolic phosphoprotein that has an important but, as yet, undefined role in cell proliferation and differentiation. Induction of growth arrest and differentiation of HL60 cells to monocytes by phorbol 12-myristate 13-acetate is associated with rapid phosphorylation of the protein. Stathmin phosphorylation was not seen when HL60 cells were induced to differentiate to monocytes, by 1 alpha, 25-dihydroxyvitamin D3, and to neutrophils, by all-trans retinoic acid and granulocyte colony stimulating factor. In all the above instances, stathmin expression was down-regulated. Thus, increased stathmin phosphorylation is not required for cell growth arrest or differentiation or down-regulation of stathmin expression.

Stathmin is expressed by the proliferating hepatocytes during liver regeneration.

Aim-To determine the liver cell populations that express the phylogenetically conserved cytosolic protein stathmin during liver regeneration.Methods-Double immunostaining for stathmin and the Ki67 antigen was performed on sections of formaldehyde fixed, paraffin wax embedded tissues from 31 liver specimens. These included a variety of disease conditions characterised by some degree of hepatocyte regeneration. Quantitative western blot analysis was performed on 22 of these specimens.Results-Variable amounts of stathmin protein were detected by western blotting in all of the specimens examined. Stathmin was not detected in three cases of histologically normal liver. On immunostaining, stathmin was demonstrated in a proportion of hepatocytes as well as lymphoid inflammatory cells and other tissue elements. In all cases most of these stathmin positive cells showed nuclear positivity for the Ki67 antigen.Conclusions-Stathmin is expressed by proliferating hepatocytes but not by resting hepatocytes. Thus, it is likely that the protein has a function important to cell proliferation as opposed to cell differentiation.

Stathmin expression is a feature of proliferating cells of most, if not all, cell lineages.

BACKGROUND: Stathmin is a phylogenetically conserved protein which was identified initially as a prominent cytosolic protein in hemopoietic cells, endocrine cells, brain, and testis. In these tissues, it has been suggested that the level of stathmin expression is important in development and cell proliferation. Furthermore, stathmin phosphorylation appears to be involved in the regulation of cell growth arrest, terminal differentiation, and hormone secretion. Elevated levels of expression of stathmin have been described in leukemia and lymphoma cells. The aim of this study was to characterize the distribution of cells that express the stathmin protein in a wide variety of normal human and rodent tissues. EXPERIMENTAL DESIGN: First, antisera against a synthetic stathmin peptide have been raised in rabbits and the specificity of these antisera confirmed by their reactivity with stathmin on 1-dimensional and 2-dimensional Western blots. Second, the most appropriate means of fixing tissues in order to stain stathmin has been investigated. Finally, using the optimized conditions of tissue fixation, the antisera have been used to immunostain sections taken from a wide variety of tissues. RESULTS: Immunopositivity was found in cells of all the lineages studied, with the stained cells present within the proliferating compartment of tissues. Conversely, most nonproliferating mature cells did not stain with the antisera to stathmin. The only nonproliferating cells that appeared to express stathmin were a subpopulation of glial cells, neurons, and anterior pituitary cells. CONCLUSIONS: It is proposed that stathmin is necessary for cell proliferation in most, or all, cell lineages and that its primary function relates to some aspect of cell division.

Persistent growth of BALB/C mouse plasmacytoma and human myeloma cell lines in the presence of phorbol myristate acetate is associated with continued expression of Lap18 (stathmin).

Lap18 is a highly conserved cytosolic protein that is expressed in dividing cells. Data from a number of studies show that a range of cell lines and mitogen-stimulated normal cells cultured in PMA phosphorylate and subsequently down-regulate Lap18. This has been found to be associated with growth arrest, although it is not clear that these events are causally related. In the present study we confirm that the HL60 promyelocytic leukemia and K562 erythroleukemia cell lines, when cultured with PMA, behave in this manner. This was not the case for any of five mouse plasmacytoma cell lines and six lines derived from patients with multiple myeloma or plasma cell leukemia. All of these lines contain Lap18, although the level of this protein in the mouse but not the human plasmacytoma cell-line cells is relatively low. All the neoplastic plasma cell-line cells phosphorylate Lap18 on culture with PMA, but this does not induce growth arrest nor result in down-regulation of Lap18 expression. Further experiments are required to test whether there is a mechanistic relationship between the continued growth of plasmacytoma cell lines and their failure to down-regulate Lap18 on culture in PMA.