Subcellular Localization of Proteins Responding to Mitoxantrone-Induced DNA Damage in Leukaemic Cells.

The aim of the present study was to investigate the subcellular localization of proteins participating in the double-strand break response pathway - p53, Mdm2, p21 and Chk2. MOLT-4 cells were pre-treated with mitoxantrone in concentrations 1 nmol/l and 5 nmol/l. The trypan blue technique was used to determine cell viability and proliferation. Western blotting was used to evaluate changes in p53, Mdm2 and Chk2 protein expression and sandwich ELISA was used to evaluate changes in the p21 protein amount. After 1 nmol/l mitoxantrone cells did not die, but their ability to proliferate was decreased. The p53 protein was activated and phosphorylated at serines 15 and 392 and accumulated in the nucleus after 24 and 48 h. The Mdm2 protein was present in the cytoplasm with its maximal level after 8 and 16 h. The p21 protein was detected in the nucleus after 24 and 48 h. Increased levels of phosphorylated Chk2 at threonine 68 were observed in the cytoplasmic fraction after 24 and 48 h of mitoxantrone treatment. We used mitoxantrone as an inducer of double-strand breaks to bring new data about the subcellular distribution of proteins responding to DNA damage. In MOLT-4 cells, the p53 protein was activated. p53 was phosphorylated at serines 15 and 392 and accumulated in the nucleus. The Mdm2 protein was activated in advance to p53 and occurred in the cytoplasm. The p21 protein was present in the nucleus. Chk2 kinase was activated by the phosphorylation at threonine 68 and we observed increased levels of this protein in the cytoplasmic fraction.

Experts and performance in histopathology--a study in breast pathology.

This study was undertaken to determine if it was possible to identify expertise within Histopathologists (trainees, district general pathologists and pathologists with a special interest in breast disease) using an objective measure of performance. The method of assessment of performance is based on the CWS (Cochran-Weiss-Shanteau) ratio formed by the individual's ability to discriminate between a spectrum of disease categories and their level of inconsistency when assessed at intervals. The slides circulated represented the spectrum of breast disease seen in routine practice. The results demonstrated the average CWS ratio to be lowest in trainees and highest in pathologists with a special interest in breast pathology although there was no statistical difference in the CWS scores obtained between the district general pathologists and pathologists with a special interest. Differences in inconsistency rather than discriminatory ability mainly accounted for the difference in the CWS ratio observed between the groups studied. The study shows that the CWS ratio is potentially a very useful tool in the assessment of pathologists with regard to assessing their progress through training.

Immunocytochemical quantitation of inflammatory cells associated with Helicobacter pylori infection.

Inflammatory cells in Helicobacter pylori-associated gastritis (HAG) were quantified. The results were compared with the gastric biopsy samples from patients with non-Helicobacter pylori gastritis (non-HAG). In order to detect immunocytochemically sensitive antigens, the biopsy samples were acetone-fixed at -20 degrees C and processed into glycol methacrylate at 4 degrees C. Parallel routine biopsy samples were formalin-fixed and paraffin-embedded. A larger, more selective retrospective study was also established examining routinely processed gastric biopsy samples. The results of both studies were expressed as numbers of individual cell types/mm2 of gastric mucosa, and showed that neutrophil granulocytes were significantly increased in HAG whereas T-lymphocytes were not. Heli. pylori colonisation was associated with a significant increase of CD8-positive lymphocytes in the epithelium of the crypts.

Thrombotic thrombocytopenic purpura mimicking acute small bowel Crohn's disease.

A 39 year old woman presented with a short history of bloody diarrhoea. She subsequently developed microangiopathic haemolysis, platelet consumption, and renal impairment. Initial investigations suggested underlying Crohn's disease of the terminal ileum complicated by sepsis and disseminated intravascular coagulation. However, after resection of a perforated caecum and terminal ileum, the diagnosis of thrombotic thrombocytopenic purpura was made. There was weak serological evidence of yersinia infection, this may have caused the early localisation of the lesions to the terminal ileum. This is believed to be the first report of thrombotic thrombocytopenic purpura affecting the small bowel alone at presentation.

Liver lesions in BALB/C mice induced by an anabolic androgen (Decadurabolin), with and without pretreatment with diethylnitrosamine.

Male BALB/C mice were treated with the anabolic androgen Decadurabolin (17 beta-hydroxy-19-nor-androst-4-ene-17 beta-n-decanoate) (DecaD) with and without pretreatment with diethylnitrosamine (DEN). Treatment with DEN alone caused development of severe hepatocytic dysplasia in all animals; 11/11 had cystadenomas and 3/11 haemangiomas, both of which were benign. Mice treated with DecaD alone developed only moderate to severe hepatocytic dysplasia. Treatment with DecaD after DEN induced neoplasms in the livers; 15/15 had severe dysplasia; 14/15 had cystadenomas, 2/15 had haemangiomas, and 4/15 developed nodular proliferation. Single hepatomas were seen in 4/15 livers, two of which also had multiple hepatomas. Control animals had normal liver histology. We conclude that DecaD enhances the hepatocarcinogenicity of DEN, especially with respect to neoplastic lesions in hepatocytes.

The effects of long-term administration of methyltestosterone on the development of liver lesions in BALB/c mice.

Male and female BALB/c mice were treated orally with 5 mg/kg body weight of methyltestosterone for 10 months. Age-matched control animals received no steroid treatment. The livers of all animals were examined by standard histopathological methods. Hepatocellular dysplasia was found in livers of all treated mice, but not in control animals. The extent and severity of this dysplasia showed a sex difference. Most of the male mice (55/78) developed severe dysplasia and in 23/78 the dysplasia was only moderate; the corresponding numbers for female mice were: mild, 15/71; moderate, 48/71 and severe dysplasia, 8/71. Microscopic hepatocytic nodules were found in 6/78 males and 7/71 females. Hepatocellular carcinoma was seen in one male mouse. These findings indicate that methyltestosterone appears to act as a weak total hepatocarcinogen under the experimental conditions used.

Familial dermal cylindroma with involvement of the parotid gland.

The association of turban tumour with other dermal cylindromas in the parotid gland is very rare. Another case is reported in which the hereditary nature of the disease is discussed and certain aspects of the treatment and management of these cases are considered.

Nodular regenerative hyperplasia of the liver with membranous glomerulonephritis.

Nodular regenerative hyperplasia of the liver is an uncommon disorder, the main consequence of which is portal hypertension, hepatocellular decompensation being uncommon. A case associated with membranous glomerulonephritis is reported, in which death was caused by fulminant hepatic failure.

Intrahepatocytic copper and copper-binding protein in primary biliary cirrhosis. Discrimination from chronic active hepatitis by means of orcein and rubeanic acid techniques.

Liver tissue samples from 111 patients with various hepatic disorders including 37 with primary biliary cirrhosis (PBC) and 22 with chronic active hepatitis (CAH) were studied. Of the PBC patients, 33 out of 37 (89%) showed orcein-positive and rubeanic acid-positive intrahepatocytic copper granules. In the 4 negative cases conventional staining methods revealed characteristic bile duct lesions and granuloma formations; all 4 were so-called presymptomatic patients. Amongst the liver biopsy samples from CAH patients, 12 out of 22 (55%) contained orcein-positive granules, but none showed stainable copper. We conclude that (1) combined orcein and rubeanic acid positivity in liver samples helps to distinguish between PBC and CAH; (2) in presymptomatic PBC the negativity of both methods is adequately compensated by a high frequency of characteristic bile duct lesions and granuloma formation: (3) the absence of orceinophilic intrahepatocytic granules, in our hands, does not accurately predict the response to steroid therapy.

Histological demonstration and frequency of intrahepatocytic copper in patients suffering from alcoholic liver disease.

This study was carried out to determine whether stainable copper accumulates in the hepatocytes of patients with alcoholic liver disease. Liver sections from 44 cases of alcoholic cirrhosis and 32 cases of non-cirrhotic alcoholic liver disease were stained by the rubeanic acid, rhodanine and orcein methods. Intrahepatocytic copper granules were found in 13 cases of cirrhosis (30%), but in none of the non-cirrhotic livers. The abundance of granules did not appear to be related either to the activity of the cirrhotic process or to the presence of cholestasis. It may well be that alcoholic cirrhosis is the most common disease associated with excess of intrahepatocytic copper.

Miliary tuberculosis in association with chronic cutaneous tuberculosis.

A case of miliary tuberculosis associated with two different forms of cutaneous tuberculosis and a negative Heaf tuberculin test is reported. The value of skin biopsy and culture in helping to establish an early diagnosis is discussed. The unusual finding of a negative Heaf tuberculin test is commented upon.

Endoscopic appearances and histological changes in ulcer-associated duodenitis.

A simple system of grading the endoscopic appearances of duodenitis is described. There was a good association between the degree of duodenitis recorded using this method and that obtained from a histological grading system in a group of duodenal ulcer patients and a group of dyspeptic patients with no endoscopic evidence of duodenitis. Endoscopic assessment is recommended as a reliable guide to the presence of duodenitis. After 1 month of treatment with cimetidine there was a significant improvement in the endoscopic duodenitis scores of a group of 17 duodenal ulcer patients. In 16 of these patients there was complete ulcer healing. Duodenitis can be a manifestation of chronic duodenal ulcer disease. In the absence of ulceration, the endoscopic diagnosis of duodenitis is of value in the recognition of patients with this condition.