bluuebunny/crossref_metadata_2025 huggingface dataset:
Clinical importance of squamous metaplasia in invasive transitional cell carcinoma of the bladder.
Authors: Martin J E , Jenkins B J , Zuk R J , Blandy J P , Baithun S I
Published: June 2007

One hundred cases of transitional cell carcinoma of the bladder were studied to determine whether squamous metaplasia and other histological features within the bladder can be of value in predicting outcome of treatment with radiotherapy. Sixty cases showed the changes of squamous metaplasia, and of this group 46 (78%) failed to respond to radiotherapy. A significant response rate of 90% was seen in the 40 tumours without squamous metaplasia. It is concluded that transitional cell carcinomas of the bladder showing squamous metaplasia are mainly resistant to radiotherapy and alternative treatment methods should be sought.
Clinical importance of DNA content in rectal cancer measured by flow cytometry.
Authors: Jass J R , Mukawa K , Goh H S , Love S B , Capellaro D
Published: June 2007

The DNA content of 369 rectal cancers was measured by flow cytometry. One hundred and four (28%) were diploid, 252 (68%) were aneuploid, and 13 (3.5%) were tetraploid. Diploid cancers were associated with an improved 5 year survival (p less than 0.001) and were more likely to present at an early stage. DNA content, however, did not confer independent prognostic information in a Cox model based on four discrete pathological variables. Patients were classified by a new system of prognostic grouping and those with a very good or a very poor outlook were removed leaving 137 prognostic group III patients. No further substratification of this group by DNA content or by four additional pathological variables could be achieved. As the new prognostic system is not improved by the addition of ploidy, routine adoption of flow cytometry in the assessment of rectal cancer cannot be recommended.
Morphometric analysis of suprabasal cells in oral white lesions.
Authors: Shabana A H , el-Labban N G , Lee K W , Kramer I R
Published: June 2007

Surgical specimens from the cheek mucosa of 73 patients with white lesions were studied to determine various morphometric parameters that would help differentiate between the various types of oral mucosal white lesions that carry a risk of malignant change. Four cell types were represented: traumatic keratosis, leucoplakia, candidal leucoplakia and lichen planus, in addition to a control group of normal mucosa. The shape and size of the epithelial cells in two cell compartments, parabasal and spinous, were investigated by an interactive image analysis system (IBAS-1). The results showed an increase in the cell size in the parabasal cell compartment of all the white lesions compared with the normal mucosa. In the spinous cell compartment there was an increase in the cell size in lichen planus and traumatic keratosis; leucoplakia and candidal leucoplakia showed a slight decrease in cell size compared with the normal mucosa. Attempts to discriminate between the four groups of white lesions showed that these parameters can provide a high level of separation between lichen planus and the three other groups, but not between leucoplakia, candidal leucoplakia, and traumatic keratosis.
Jejunal enteropathy associated with human immunodeficiency virus infection: quantitative histology.
Authors: Batman P A , Miller A R , Forster S M , Harris J R , Pinching A J , Griffin G E
Published: June 2007

Jejunal biopsy specimens from 20 human immunodeficiency virus (HIV) positive male homosexual patients were analysed and compared with those of a control group to determine whether the abnormalities were caused by the virus or by opportunistic infection. The degree of villous atrophy was estimated with a Weibel eyepiece graticule, and this correlated strongly with the degree of crypt hyperplasia, which was assessed by deriving the mean number of enterocytes in the crypts. The density of villous intraepithelial lymphocytes fell largely within the normal range, either when expressed in relation to the number of villous enterocytes or in relation to the length of muscularis mucosae. Villous enterocytes showed mild non-specific abnormalities. Pathogens were sought in biopsy sections and in faeces. Crypt hyperplastic villous atrophy occurred at all clinical stages of HIV disease and in the absence of detectable enteropathogens. An analogy was drawn between HIV enteropathy and the small bowel changes seen in experimental graft-versus-host disease. It is suggested that the pathogenesis of villous atrophy is similar in the two states, the damage to the jejunal mucosa in HIV enteropathy being inflicted by an immune reaction mounted in the lamina propria against cells infected with HIV.
Histopathological assessment of bleeding from polyps of the colon and rectum.
Authors: Waldock A , Ellis I O , Armitage N C , Turner D R , Hardcastle J D
Published: June 2007

One hundred and twenty seven colorectal polyps were examined to assess histopathological evidence of recent and old haemorrhage to test the usefulness of faecal occult blood tests in detecting colorectal neoplasia, in particular premalignant adenomas. Evidence of haemorrhage was consistently found in adenomas but was rare in non-neoplastic polyps. Haemorrhage within adenomas was predominantly stromal and associated with dilated, congested vessels. Factors associated with more severe haemorrhage were size, pedunculation, and villous growth; the degree of epithelial dysplasia and the age and sex of the patient were not associated factors. Proximal polyps showed more old haemorrhage than rectal polyps, but there was no such difference for recent haemorrhage.
Histopathology of breast cancer in young women in relation to use of oral contraceptives.
Authors: Miller N , McPherson K , Jones L , Vessey M
Published: June 2007

A detailed review was made of the histopathology of 227 tumours taken from 261 women under 45 years of age with breast cancer. The tumours were classified as follows: whether oral contraceptives had been used at any time; and whether oral contraceptives had been used before first term pregnancy. All analyses were adjusted for the effects of age. Overall, 201 (88.5%) of tumours were infiltrating ductal in type, 19 (8.4%) were infiltrating lobular, and seven (3.1%) were in situ ductal. Of the infiltrating ductal tumours, 28 (14%) were grade I 88 (44%) grade II, and 84 (42%) grade III. Various other tumour characteristics were also examined (per cent of carcinoma in situ, lymphatic permeation, necrosis, lymphoplasmocytic reaction and tumour edge). None of the histopathological features assessed showed any significant association with oral contraceptive use. Some characteristics of the areas of breast tissue adjacent to the tumours were also studied. The degree of cyst formation was considerably less pronounced in those using oral contraceptives before first term pregnancy than in those not doing so.
Bone histology in young adult osteoporosis.
Authors: Hills E , Dunstan C R , Wong S Y , Evans R A
Published: June 2007

Bone histology was quantitated in 10 osteoporotic patients aged between 17 and 51 years and in six healthy subjects aged between 23 and 43 years. The osteoporosis was of varying aetiology and was clinically stable. All patients were given tetracycline before biopsy and double tetracycline labelling was used in seven patients. Bone forming and resorbing surfaces were defined by the presence of osteoblasts and osteoclasts, respectively, which were identified by histochemical techniques. The associations between bone forming and resorbing surfaces were similar in patients and controls, though the range of values was wider in the patients than in the controls. Mineral apposition rate was normal in the osteoporotic patients, but there was a reduction in mineralising (tetracycline) surface, whether related to osteoid surface or to osteoblast surface. This did not indicate osteomalacia as the directly and indirectly measured mineralisation lag times were normal. The osteoid seams were thinner in osteoporotic patients than in controls. The data suggest that osteoclast and osteoblast numbers were normal in this group of osteoporotic patients but that the metabolic activity of osteoblasts was impaired.
KP1: a new monoclonal antibody that detects a monocyte/macrophage associated antigen in routinely processed tissue sections.
Authors: Pulford K A , Rigney E M , Micklem K J , Jones M , Stross W P , Gatter K C , Mason D Y
Published: June 2007

A new monoclonal antibody, KP1, raised against a lysosomal fraction of human lung macrophages, recognises a fixation-resistant epitope in a wide variety of tissue macrophages (such as Kupffer cells germinal centre, splenic, and lamina propria macrophages), and in granulocyte precursors. Its broad reactivity with cells of the mononuclear phagocytic lineage was established by testing on routinely processed samples of normal and reactive lymphoid tissues. Interdigitating reticulum cells were unstained or showed limited cytoplasmic staining while Langerhans' cells and follicular dendritic reticulum cells were unreactive. KP1 recognises a molecule of about 110 kilodaltons in macrophage-rich human tissue when tested by either immunoprecipitation or Western blotting (although the latter procedure also shows two additional components with molecular weights of 70 and 40 kilodaltons). KP1 should be of considerable value for studying disorders of the monocyte/macrophage system, including both reactive and neoplastic states (such as true histiocytic proliferations).
Cell suspensions from collagenase digestion of bone marrow trephine biopsy specimens.
Authors: Ades C J , Ablett G A , Collins R J , Bunce I H
Published: June 2007

A technique for the extraction of cells from bone marrow trephine core biopsy specimens using collagenase digestion was assessed in 39 cases (33 diagnostic and six normal). Diagnostically useful numbers of cells were extracted from all marrows. Morphological assessment of cytocentrifuge preparations of these cells gave a correct diagnosis in 23 (60%) of cases compared with 27 (70%) for the corresponding aspirated marrow smears. Phenotypic analysis using flow cytometry showed persistence of a range of surface membrane antigens following collagenase digestion. Increased autofluorescence was a problem in some cases. Cytochemistry, bone marrow culture, and cytogenetic analysis could also be carried out on these cells. It is concluded that this technique has useful diagnostic applications in cases of dry taps.
Sclerosing cholangitis with hepatic microvesicular steatosis in cystic fibrosis and chronic pancreatitis.
Authors: Benett I , Salh B , Haboubi N Y , Braganza J M
Published: June 2007

The association between asymptomatic primary sclerosing cholangitis and exocrine pancreatic disease was underlined by the findings in a patient with cystic fibrosis and in another with chronic pancreatitis. In each case hepatocytes showed extensive microvesicular steatosis and studies of drug metabolism suggested hepatic enzyme induction: biliary or serum analysis, or both, disclosed raised concentrations of a lipid-based marker of free radical oxidation. These findings suggest that toxic metabolites of oxygen or other chemicals may have a role in the pathogenesis of the bile duct lesion.
Infections in British clinical laboratories, 1986-87.
Authors: Grist N R , Emslie J A
Published: June 2007

During 1986-87 this continuing survey showed 15 specific infections in the staff of 235 laboratories, representing 28,524 person years of exposure. The community was the probable source of four of the five cases of tuberculosis and one of the five cases of salmonellosis. Occupational exposure was the probable cause of four infections by Shigella flexneri, three by Salmonella typhimurium, and one by S typhi, all affecting medical laboratory scientific officers (MLSOs) in microbiology. Occupational exposure was also the probable cause of one case of tuberculosis in a mortuary technician and one of probable non-A, non-B hepatitis in a medical laboratory scientific officer haematology worker. The overall incidence of reported infections was 52.6/100,000 person years (35/100,000 for infections of probable occupational origin). The highest rates of laboratory acquired infections related to MLSO microbiology workers and mortuary technicians. No additional infections were seen as a result of extending the survey to forensic laboratories.
Quantity of nuclear DNA in malignancies and benign lymphadenopathies associated with Epstein-Barr virus.
Authors: Lehtinen T , Lehtinen M , Aine R , Dammert K , Kulomaa P , Alavaikko M , Leinikki P
Published: June 2007

The quantity of nuclear DNA in 90 tumours with a strong probability of being associated with Epstein-Barr virus (EBV) (11 cases with nasopharyngeal carcinomas, seven cases of endemic and 26 of non-endemic Burkitt's lymphoma, and 46 cases of Hodgkin's disease) were analysed by flow cytometry. Twenty three cases with benign lymphadenopathies were analysed in a similar way. Except for endemic Burkitt's lymphoma most of the tumours were diploid. Near-diploid aneuploidy (with a DNA index ranging from 1.06 to 1.26) was also found in endemic Burkitt's lymphoma as well as in six non-endemic Burkitt's lymphomas and in eight cases of Hodgkin's disease but was absent in nasopharyngeal carcinomas. Tetraploid aneuploidy was seen in three cases of nasopharyngeal carcinoma. Five of the 23 cases of lymphadenopathy also showed near-diploid DNA, one of which subsequently developed into a non-Hodgkin's lymphoma. It is concluded that near-diploid DNA content seems to be associated with the lymphatic origins of the tumours rather than with EBV.
Abnormal platelet aggregation in patients with Raynaud's phenomenon.
Authors: Biondi M L , Marasini B
Published: June 2007

Platelet aggregation in vitro to several aggregating agents (serotonin (5-HT), adenosine diphosphate, adrenaline and collagen) was studied in 16 patients with primary and secondary Raynaud's phenomenon and compared with that in 13 normal volunteers. Platelets from patients with Raynaud's phenomenon had significantly greater responses to all the 5-HT concentrations tested (p less than 0.001 for 10 microM; p less than 0.01 for 1 microM; p less than 0.05 for 0.1 microM; p less than 0.02 for 0.025 microM) and to low doses of adenosine diphosphate (p less than 0.01 for 1 microM; p less than 0.02 for 0.5 microM) but normal responses to collagen, adrenaline, and high doses of adenosine diphosphate. Patients with secondary Raynaud's phenomenon were significantly more hypersensitive to 0.5 microM adenosine diphosphate than patients with primary Raynaud's phenomenon. In patients with secondary Raynaud's phenomenon there was a significant correlation between the extent of 5-HT aggregation and the duration of the disease. The finding that platelets from patients with Raynaud's phenomenon have enhanced responses to 5-HT and adenosine diphosphate, but normal responses to adrenaline and collagen, is consistent with a role for 5-HT in this disease.
Efficient selection of tests for bacteriological typing schemes.
Authors: Gaston M A , Hunter P R
Published: June 2007

To simplify the selection of tests for bacteriological typing methods, such as bacteriophage, bacteriocin, and biotyping, a computerised method was assessed. This uses a numerical index of discrimination (D) to facilitate the selection of an efficient typing set. The computer programs take the most discriminatory test as the initial test in the partial typing set, and then select the next test by combining each of the remaining candidates with the partial set and choosing the test which maximises D. This cycle is repeated until the remaining candidates do not increase the discriminatory power of the typing set. Options are provided for the investigator to pre-select certain tests for inclusion or exclusion from the typing set. It is concluded that the numerical index D is a simple means of test selection, but it must be emphasised that it is important to combine its use with data on the incidence of reaction in each test, on reproducibility, and on the similarity among tests.
In vitro amplification of hepatitis B virus sequences from liver tumour DNA and from paraffin wax embedded tissues using the polymerase chain reaction.
Authors: Lo Y M , Mehal W Z , Fleming K A
Published: June 2007

A 185 base pair fragment from the core-polymerase overlap region of the hepatitis B virus (HBV) genome was amplified using the polymerase chain reaction (PCR). The results were compared with those of Southern blotting on extracted DNA from eight hepatocellular carcinomata. The data agreed with those of Southern blotting in six cases (two positive, four negative) but in two other positive cases PCR failed to amplify HBV sequences. This suggests deletion or mutation, or both, of this viral region in these cases. PCR was also used to amplify HBV sequences from formalin fixed, paraffin wax embedded tissue. Tissue inhibition of PCR occurred which increased with the number of tissue sections. It was present in tissues from different organs and species and fixed by different procedures, thus highlighting the need for a positive control during amplification. Use of formalin fixed Alexander cells, however, showed a sensitivity of one viral copy per 5000 cells. Confirmation of the identity of the PCR products was carried out using PCR-generated biotinylated probes, and suggested the insertion of extra nucleotide sequences or infection with an HBV variant in one case.
New method for detecting slime production by coagulase negative staphylococci.
Authors: Freeman D J , Falkiner F R , Keane C T
Published: June 2007

An alternative method for detecting the production of slime by coagulase negative staphylococci was compared with the routinely used Christensen method on 124 isolates of coagulase negative staphylococci from carriage sites, blood cultures, and infected peritoneal dialysis fluids. The alternative method requires the use of a specially prepared solid medium--brain heart infusion broth, supplemented with 5% sucrose, and Congo red stain. Of the 124 tests, there was complete agreement between methods in 107 and only one strain was clearly negative by Christensen's method while positive on Congo red agar. The Congo red method is rapid, sensitive, and reproducible and has the advantage that colonies remain viable on the medium. It is also not subject ot interbatch variation of media which sometimes affects the reproducibility of the Christensen method.
Improved method for immunoperoxidase detection of membrane antigens in frozen sections.
Authors: Green M A , Sviland L , Malcolm A J , Pearson A D
Published: June 2007

The visualisation of membrane antigens in frozen sections using metal-enhanced 3,3'diaminobenzidine (DAB) was investigated. Particular attention was paid to the degree of reaction enhancement, the disruptive effect on morphological detail, and the ease of the techniques. The best result was obtained using nickel-modified DAB at pH 6.0 to develop the peroxidase enzyme, with further enhancement in cobalt chloride at neutral pH. Silver methenamine enhancement is also possible but can give rise to non-specific or background staining.
Postmortem audit in a paediatric cardiology unit.
Authors: Russell G A , Berry P J
Published: June 2007

Postmortem examinations performed on 76 children with a clinical diagnosis of congenital heart disease were reviewed retrospectively and compared with the findings before death. Both operated and unoperated cases were studied over a three year period. Despite intensive investigation during life, there was a high rate of unsuspected abnormalities at necropsy (80%): 29 cases had undiagnosed additional cardiac anomalies or surgical flaws, which contributed to death in 13 cases. Defects in surgery were uncommon but permitted modification in surgical technique to avoid recurrence. Myocardial necrosis and pulmonary foreign body embolism were common findings, the importance of which is uncertain and requires further study for their prevention. Even in the most thoroughly investigated cases postmortem examination has a high yield of clinically important pathology which is undetected during life.
Gastric mucosa in patients with portal hypertension: prevalence of capillary dilatation and Campylobacter pylori.
Authors: Foster P N , Wyatt J I , Bullimore D W , Losowsky M S
Published: June 2007

To determine whether the "congestive" gastropathy associated with portal hypertension showed distinctive histological features independent of inflammatory gastritis, endoscopic biopsy specimens of gastric mucosa from 23 patients with portal hypertension and 25 patients with non-ulcer dyspepsia were examined. Active chronic gastritis associated with Campylobacter pylori was found in three patients with portal hypertension compared with 13 patients with non-ulcer dyspepsia. The changes of reflux gastritis were seen in nine patients with portal hypertension compared with three patients with non-ulcer dyspepsia. Mucosal capillary dilatation, assessed on sections stained for factor VIII related antigen, a specific marker for endothelial cells, was significantly greater in biopsy specimens from patients with portal hypertension but this difference was not apparent on sections stained conventionally. The degree of capillary dilatation was unrelated to the presence of histological gastritis. These observations support the view that portal hypertension is associated with a distinctive gastropathy characterised by prominence and dilatation of mucosal capillaries.
Expression of laminin in benign and malignant sclerosing lesions of extrahepatic bile ducts.
Authors: Haglund C , Roberts P J , Nordling S
Published: June 2007

The distribution of laminin, a basement membrane glycoprotein, was studied by an immunohistochemical technique in six samples of benign sclerosing lesions of extrahepatic bile ducts and in 11 sclerosing cholangiocarcinomas. The expression of laminin showed that benign glandular structures were surrounded by a mainly intact basement membrane. In sclerosing cholangiocarcinomas laminin was irregularly distributed, and in large areas totally absent. The findings suggest that deposition of basement membrane material is defective. Sclerosing cholangitis may be extremely difficult to distinguish from well differentiated sclerosing cholangiocarcinoma. This sample was small, but the diseases studied were relatively rare. Staining for laminin might be of additional use to surgical pathologists in the differential diagnosis of benign sclerosing lesions and sclerosing cholangiocarcinomas.
Angiolymphoid hyperplasia with eosinophilia: possible aetiological role for immunisation.
Authors: Hallam L A , Mackinlay G A , Wright A M
Published: June 2007

Five young children (mean age 26.4 months) with angiolymphoid hyperplasia with eosinophilia (Kimura's disease) from either the upper arm or buttock were identified over 18 months. The unusual distribution of the lesions and the young age of the patients suggested a possible association with immunisation. The clinical and histopathological features in these cases were accordingly reviewed. The biopsy specimens showed the usual histological appearances of a prominent inflammatory component, fibrosis, and vascular proliferation associated with aggregates of eosinophils. The features were those of a reactive rather than neoplastic process. Immunohistochemical preparations showed positive staining of variable numbers of plasma cells with antibodies to IgG, IgM, IgA and IgE and a reticular staining of germinal centres with IgM and IgE antibodies. Immunisation histories obtained from the patients' general practitioners showed a remarkable correlation between the distribution of the lesions and the sites of injections and an aetiological role for immunisation in these cases seems likely.
Leukaemic phase of mantle zone (intermediate) lymphoma: its characterisation in 11 cases.
Authors: De Oliveira M S , Jaffe E S , Catovsky D
Published: June 2007

Sixteen patients presented with B cell leukaemia (white cell count 26-269 x 10(9)/l) which could not be classified as chronic lymphocytic (CLL), prolymphocytic leukaemia, or follicular lymphoma in leukaemic phase. Eleven patients (10 men, one woman) corresponded histologically to intermediate (INT) or mantle zone lymphoma, and five, with less well defined features, were designated small lymphocytic lymphoma with cleaved cells. The blood films showed a pleomorphic picture with lymphoid cells of predominantly medium size with nuclear irregularities and clefts. The membrane phenotype of the circulating cells showed strong immunoglobulin staining and reactivity with CD5 and FMC7 in all cases tested; CD10 was positive in six out of nine cases. The membrane phenotype of two of the five cases of small lymphocytic lymphoma was close to those of B-CLL and three resembled INT lymphoma. Bone marrow trephine biopsy specimens showed a diffuse pattern of infiltration in INT lymphoma. The median survival of these patients was less than two years, suggesting that a leukaemic presentation is associated with poor prognosis. By combining data from histology, membrane markers, and peripheral blood morphology, the leukaemic phase of typical INT lymphoma can be defined in most cases.
Antibodies to Escherichia coli O157 in patients with haemorrhagic colitis and haemolytic uraemic syndrome.
Authors: Chart H , Scotland S M , Smith H R , Rowe B
Published: June 2007

Twenty four sera from patients with haemolytic uraemic syndrome or haemorrhagic colitis and healthy controls were examined for antibodies to the lipopolysaccharide (LPS) of Escherichia coli O157. Faecal specimens from these patients were also examined for Vero cytotoxin producing E coli (VTEC) by DNA probes, and for faecal Vero cytotoxin. Eight patients with faecal E coli O157:H7 gave a strong antibody response to O157 LPS, shown by immunoblotting and an enzyme linked immunosorbent assay. Six symptomatic patients without evidence of faecal VTEC also gave a strong antibody response to O157 LPS. Sera from the remaining five patients and five healthy controls did not contain antibodies to E coli O157. The results suggest that the testing of sera from patients with haemorrhagic colitis or haemolytic uraemic syndrome by ELISA or immunoblot would prove valuable in addition to the established procedures for detecting VTEC, using DNA probes and testing for faecal Vero cytotoxin.
Chronic carotid glomitis and the glomus pulmonale.
Authors: Khan Q , Heath D
Published: June 2007

A histological study was made of the carotid bodies and glomus pulmonale in 50 consecutive subjects over the age of 50 years who came to necropsy to determine if chronic glomitis is confined to the carotid bodies or whether it also occurs in other glomera. Lymphoid aggregates and plasma cell activity were found in the glomus pulmonale just as they were in the carotid bodies. Chronic pulmonary glomitis sometimes occurred in association with chronic carotid glomitis and sometimes independently of this. The mean age of the affected subjects was 76 years in isolated pulmonary glomitis and 79 years in chronic carotid glomitis. Chronic carotid glomitis affected seven (14%) subjects and predominated in women (six to one). In the seven cases of isolated pulmonary glomitis women predominated five to two. Chronic glomitis seems to be a disease process which may affect at least two members of the non-chromaffin paraganglionic system, but it is not clear why focal chronic inflammation affects the glomus pulmonale of one person but not of another.
Demonstration of multiple HPV types in normal cervix and in cervical squamous cell carcinoma using the polymerase chain reaction on paraffin wax embedded material.
Authors: Griffin N R , Bevan I S , Lewis F A , Wells M , Young L S
Published: June 2007

The prevalence of human papilloma virus (HPV) types 6, 11, 16 and 18 was investigated using the polymerase chain reaction on formalin fixed, paraffin wax embedded material in 19 cases of cervical squamous cell carcinoma and in 10 normal cervices. HPV DNA was detected in 16 of 19 carcinomas, with multiple types present in 11 of these. HPV 16 or 18, or both, were present in all cases in which HPV was shown. Six of 10 cases of normal cervix contained HPV; five of these contained two or more HPV types, including HPV 16 or 18, or both. This study shows the feasibility of using the PCR on paraffin wax embedded material and indicates a high rate of carriage of multiple HPV types in both normal and neoplastic cervix.
Simple technique for detecting K1 antigen of Escherichia coli.
Authors: Devine D A , Roberts A P , Rowe B
Published: June 2007

The K1 antigen is a poor immunogen, its detection by serological means is difficult, and previously described methods using K1 specific bacteriophages require standardised suspensions of five different bacteriophages. A simple technique was developed which uses an unstandardised suspension of a single K1 specific bacteriophage applied with a 1 mm wire loop to bacteria streaked on to cystine lactose electrolyte deficient (CLED) medium. The technique correctly identified all 99 known K1 strains tested, including 14 strains negative with the serological method. Among 71 clinical isolates from urinary tract infections, the single bacteriophage method distinguished 30 K1 strains from 41 strains without this antigen. Suspensions of the bacteriophage were shown to remain fully active for at least two years when stored at 4 degrees C. It is concluded that this technique required so little in materials, time, or equipment that it could be routinely used in most laboratories.
Immunocytochemical examination of bone marrow in disseminated neuroblastoma.
Authors: Carey P J , Thomas L , Buckle G , Reid M M
Published: June 2007

To assess the usefulness of immunocytochemical analysis of bone marrow in patients with neuroblastoma, marrow smears from 33 staging procedures in 12 patients were examined using an indirect immunoalkaline phosphatase technique with monoclonal antibodies raised against human neural tissue. Marrow aspirate and trephine collagenase digest specimens from individual sites were each tested with the monoclonal antibody UJ13A and with a pool of three related antibodies. The results were compared with morphological assessment of conventionally stained aspirates and trephine specimens taken at the same time. Immunostaining suggested the presence of tumour in seven of 18 staging procedures in which conventional techniques had shown infiltration. Tumour infiltration was also suggested in four of 10 staging procedures with suspicious trephine specimens, but in none of three with relatively innocent histological and cytological features. Immunological investigation provides no additional information about the presence of infiltration if conventional microscopy has shown definite tumour. When histological appearances are suspicious, immunostaining of stored aspirate smears or collagenase digest specimens may provide evidence of infiltration. There are insufficient data to comment on the value of immunostaining when conventional techniques reveal "normal" marrow, but the impression gained from this study is that immunostaining has a limited role in the detection of metastatic neuroblastoma, which yet remains to be defined.
Application of 1 nm gold probes on paraffin wax sections for in situ hybridisation histochemistry.
Authors: Jackson P , Dockey D A , Lewis F A , Wells M
Published: June 2007

An in situ hybridisation technique that uses 1 nm immunogold reagents and silver enhancement was devised to detect biotinylated DNA viral probes in formalin fixed, paraffin wax sections of human cervix. DNA probes labelled with biotin-11-deoxyuridine triphosphate were detected after hybridisation to nucleic acid sequences by an antibiotin antibody, followed by a gold labelled secondary antibody. Silver enhancement then permitted visualisation of the signal at the light microscopic level. The method was reliable and produced less background staining than previously described methods. The signal could be enhanced by epi polarisation microscopy. Furthermore, biotinylated DNA probes may be detected directly by a 1 nm gold labelled goat antibiotin antibody without loss of labelling intensity, and this may be preferable to the longer two layer technique, previously described.
Importance of sampling method in DNA analysis of lung cancer.
Authors: Carey F A , Lamb D , Bird C C
Published: June 2007

Lung carcinomas are characterised by considerable histological variation within the tumour. The possible effects of this morphological heterogeneity on the estimation of tumour ploidy were investigated. Multiple tissue blocks were systematically taken from 20 lung tumours and analysed by flow cytometry. The routine, archival paraffin wax embedded diagnostic blocks from these cases were also analysed. Nineteen (95%) of the tumours were shown to contain aneuploid stemlines by systematic sampling, but if only one of these systematic tissue blocks had been taken from each case the incidence of DNA aneuploidy could have been as low as 45%. Only 15 (75%) tumours were aneuploid when all the routine archival blocks were analysed, but by specifically selecting tumour areas from the archival material the accuracy of this method was increased to 90%. It is concluded that tumour sampling methods are of primary importance in assessing the DNA content of lung tumours. Routine paraffin wax embedded archival tissue provides a suitable source of material for this purpose, provided that "turnover" selection is carried out.
Crypt cell proliferation and HLA-DR expression in pelvic ileal pouches.
Authors: de Silva H J , Gatter K C , Millard P R , Kettlewell M , Mortensen N J , Jewell D P
Published: June 2007

To investigate the nature of the morphological changes that occur in ileal pouches, 26 biopsy specimens from patients with functioning ileo-anal pouches (eight with pouchitis) were studied. Normal ileum (n = 10) was used as a control. Mucosal morphometry (using linear measurements), crypt cell proliferation (CCP) (using the monoclonal antibody Ki67), and epithelial HLA-DR expression (monoclonal antibody CR3/43) were assessed. CCP (expressed as the percentage of Ki67 positive nuclei for each crypt) was significantly higher in pouches with pouchitis, compared with those without, and in pouches without pouchitis compared with normal ileum. CCP values in some pouches without pouchitis approached values found in those with pouchitis. CCP was related inversely to villous height and an index of villous atrophy (VH/TMT), and directly to crypt depth. In the presence of pouchitis there was intense epithelial HLA-DR expression that extended into the crypts. In some pouches with high CCP values, but without clinically important inflammation, surface epithelial HLA-DR expression was weak and patchy. It is concluded that villous atrophy and crypt hyperplasia in ileal pouches are associated with high CCP values. These may be increased even in the absence of active inflammation, and this increase may occur as a response to the new luminal environment.
Cells immunoreactive for neuropeptide in human thymomas.
Authors: Lauriola L , Maggiano N , Larocca L M , Ranelletti F O , Ricci R , Piantelli M , Capelli A
Published: June 2007

The presence of opioid peptides, bombesin, and substance P was investigated by immunohistochemistry in tissue sections from six human thymomas. The number of immunoreactive cells seemed to vary from one case to another. Ultrastructural investigation, showing the presence of desmosomes in labelled cells, allowed these cells to be classified as epithelial lineage cells. The occurrence of cells containing neuropeptide in thymomas suggest that peptide molecules could have modulated the behaviour of this tumour, given the reported influence of these molecules on immune functions and their growth promoting activity on several cell types, including mesenchymal and epithelial cells.
Tissue preparation for simultaneous flow cytometric quantitation of tumour associated antigens and DNA in solid tumours.
Authors: van Dam P A , Watson J V , Lowe D G , Cox H , Curling M , Shepherd J H
Published: June 2007

A multiparameter flow cytometric assay for the simultaneous study of tumour associated antigens (TAA) and DNA in fresh solid tumours was devised. Cell suspensions were prepared by disaggregating unfixed solid tumour samples mechanically over a stainless steel mesh. Indirect immunofluorescence was used to identify the TAA, and DNA was stained with propidium iodide. Cell morphology was well preserved, cell clumping was negligible, and high quality indirect immunofluorescence quality indirect immunofluorescence and DNA staining were obtained. The technique is simple, rapid, and reproducible. Multiparameter assays can be developed to study prognostic indicators such as membrane oncoproteins, receptors, and multidrug resistance in solid tumours. With a suitable panel of antibodies the technique might become an aid in the differential diagnosis and biochemical diagnosis of some solid tumours.
Production of extra-cellular slime by Staphylococcus epidermidis during stationary phase of growth: its association with adherence to implantable devices.
Authors: Bayston R , Rodgers J
Published: June 2007

A method of optimising slime production produced by Staphylococcus epidermis and its quantitative assay was developed, which gave a preliminary indication of its identity and an assessment of the correlation between slime production and adherence of the organism to implants. After inducing vigorous growth in brain heart infusion broth to stationary phase, all nutrients were removed by washing and the organisms resuspended in sterile deionised water with added magnesium. After further incubation the culture was centrifuged and the supernatant reacted with alcian blue in 50 mM magnesium chloride/sodium acetate solution, and the amount of bound dye was measured spectrophotometrically at 620 nm after its resolubilisation using sodium dodecyl sulphate. Large quantities of slime were produced by some, but not all, strains. Preliminary electrophoresis of the slime showed mobility and staining similar to that of the glycosaminoglycans. Adherence was tested by growing strains in wells of tissue culture plates and aspirating the supernatant after incubation. After fixation and staining of adherent growth the amount of bound stain was determined spectrophotometrically after its elution with ethanol. In this series of organisms there was no correlation between the result of tests for adherence or production of extracellular slime, and no correlation between either of these and the clinical source of the organisms.
Human papillomavirus in oesophageal squamous cell carcinoma.
Authors: Loke S L , Ma L , Wong M , Srivastava G , Lo I , Bird C C
Published: June 2007

Thirty seven cases of oesophageal squamous cell carcinoma were studied by applying DNA slot blot analysis and in situ hybridisation using type specific probes for HPV 6, 11, 16 and 18. Cases of condyloma accuminata, cervical carcinoma, and laryngeal papilloma were used as controls. Blocks including areas of invasive carcinoma, intraepithelial neoplasia, and normal epithelium were studied in each case. No HPV genome was detectable in any of the oesophageal cases. It is concluded that these types of HPV do not have an association with oesophageal squamous cell carcinoma.
Detection and typing of human papillomavirus using the Vira Type "in situ" kit: comparison with a conventional dot blot technique.
Authors: Faulkner-Jones B E , Bellomarino V M , Borg A J , Orzeszko K , Garland S M
Published: June 2007

A new commercial kit (Vira Type "in situ", Life Technologies, Inc., Molecular Diagnostics Division, Guithersburg, Maryland, USA) for the detection of human papillomavirus (HPV) types 6, 11, 16, 18, 31, 33 and 35 in routinely processed human anogenital tissue was compared with a conventional dot blot assay for HPV 6, 11, 16 and 18. Both systems use double-stranded genomic DNA probes for the detection of type specific HPV DNA. The probes used on the dot blots were labelled with 32P and visualised autoradiographically. The Vira Type probes were labelled with biotin and visualised using a streptavidin-alkaline phosphatase conjugate with NBT-BCIP substrate. Biopsy specimens from the cervix, vagina, and vulva of 46 women were processed by both methods and compared. The histological diagnoses ranged from benign changes, to dysplasia, and invasive carcinoma. Overall, 50% of biopsy specimens were positive for HPV DNA by dot blot hybridisation; only 39% were positive by Vira Type in situ hybridisation. Three of the specimens positive by the Vira Type "in situ" kit showed no cross hybridisation and were the same HPV type as the dot blot. A further 13 showed hybridisation, but the showed cross hybridisation, but the to the dot blot results. One biopsy specimen was positive for different HPV types by the two tests and one was positive by Vira Type and negative by dot blot. Six biopsy specimens were negative by Vira Type but positive by dot blot. It is concluded that the Vira Type "in situ" kit has a similar specificity but lower sensitivity than the dot blot hybridisation method for the detection of HPV DNA.
Detection of human papillomavirus type 16 DNA in carcinomas of the palatine tonsil.
Authors: Niedobitek G , Pitteroff S , Herbst H , Shepherd P , Finn T , Anagnostopoulos I , Stein H
Published: June 2007

Twenty eight tonsillar carcinomas of various histological types were investigated for the presence of Epstein-Barr virus (EBV), cytomegalovirus (CMV), and human papillomavirus (HPV) types 6, 11, and 16 by in situ hybridisation using highly stringent procedures. In six cases an autoradiographic signal was obtained in the tumour cell nuclei with the HPV type 16 specific probe. No signal was obtained with any of the other probes. Immunohistochemical investigations with mouse monoclonal antibodies directed against the L1 protein of HPV type and a rabbit antiserum that detects common protein determinants of HPV gave negative results, thus indicating latent infection. Furthermore, a series of tonsils from controls with comparable age distribution was negative by both in situ hybridisation and immunohistology. These results indicate a possible role for HPV 16 in the aetiology of a proportion of tonsillar carcinomas.
Measurement techniques for melanoma: a statistical comparison.
Authors: Calder C J , Campbell A P , Plastow S R
Published: June 2007

Inter- and intra-observer variation in measuring the depth of invasion of malignant melanomas was assessed using three different techniques: eye-piece graticule, stage Vernier, and projection image analysis. Significant variation was found for all methods but was least pronounced with the stage Vernier. It is recommended that this should be the preferred technique for routine use.
New technique for analysing conjugated bile acids in gastric juice.
Authors: Gotley D C , Morgan A P , Cooper M J
Published: June 2007

A new technique of high performance liquid chromatography (HPLC) was developed for the analysis of conjugated bile acids in gastric juice. The assay is rapid, sensitive, and highly specific for bile acid conjugates over the range 30-10,000 mumol/l and is not affected by the presence of food. Ten patients with a variety of common upper gastrointestinal disorders underwent continuous gastric aspiration for 16 hours, including a fasting, post-prandial, and nocturnal period, and aliquots of aspirates were analysed every two hours by the HPLC technique for the six most prevalent bile acid conjugates present in human hepatic bile. Intragastric bile acid concentrations were lowest in the post-prandial period and highest in the early hours of the morning. Conjugated bile acid proportions, or profiles, varied considerably from patient to patient, but tended to remain uniform over time in individual patients. It is concluded that HPLC is superior to enzymatic techniques for the analysis of conjugated bile acids in the upper gastrointestinal tract.
Antibody coated bacteria in urine of patients with recent spinal injury.
Authors: Galloway A , Green H T , Menon K K , Gardner B P , Pemberton S , Krishnan K R
Published: June 2007

Twenty patients with an acute spinal injury were prospectively studied to assess the clinical importance of antibody coated bacteria (ACB) in the urine and the association among the different bacterial species with a positive antibody coated bacteria test. Clinical urinary tract infection was associated with a positive ACB test on 45% of occasions. Three hundred and ninety nine urine samples containing 541 bacterial isolates were assessed for the presence of ACB; 13% were found to be positive and 87% negative for ACB; 67% of urines contained a single bacterial isolate. Pseudomonas aeruginosa was most commonly associated with clinical urinary tract infection, found in 25% of episodes, followed by Proteus mirabilis (17.5%), Klebsiella sp (12.5%), and Proteus morganii (10%). Providencia stuartii, however, was most commonly associated with a positive ACB test (found in 17%). Other bacteria associated with a positive ACB test included Klebsiella sp (14%), Acinetobacter sp (12.5%), Pseudomonas aeruginosa (12%), Citrobacter sp (11.5%). A positive ACB test is not to be expected from a patient with spinal injury who has a catheter in place, and the test may provide a useful guide to identify those patients with an invasive infection. It is doubtful that a decision to treat or not treat bacteriuria could rest on the identification of the bacterial species alone.
Type II oestrogen binding sites in human colorectal carcinoma.
Authors: Piantelli M , Ricci R , Larocca L M , Rinelli A , Capelli A , Rizzo S , Scambia G , Ranelletti F O
Published: June 2007

Seven cases of colorectal adenocarcinomas were investigated for the presence of oestrogen receptors and progesterone receptors. The tumours specifically bound oestradiol. This binding almost exclusively resulted from the presence of high numbers of type II oestrogen binding sites. Oestrogen receptors were absent or present at very low concentrations. Immunohistochemical investigation of nuclear oestrogen receptors gave negative results. This indicates that antioestrogen receptor antibodies recognise oestrogen receptors but not type II oestrogen binding sites. The presence of specific type II oestrogen binding sites and progesterone binding offers further evidence for a potential role for these steroids and their receptors in colorectal carcinoma.
Comparative in situ hybridisation study of juvenile laryngeal papillomatosis in Papua New Guinea and Australia.
Authors: Wright R G , Murthy D P , Gupta A C , Cox N , Cooke R A
Published: June 2007

A comparative study of cases of juvenile laryngeal papillomatosis from Papua New Guinea (n = 3) and Brisbane, Australia (n = 9) was carried out. In situ hybridisation reactions for human papillomavirus (HPV) types 6 and 11 occurred in 11 cases. All three cases from Papua New Guinea and eight from Australia gave positive signals. A negative reaction was observed in one Australian case. The intensity of the reaction was strong in seven cases, moderate in one, and weak in three. An equivocal reaction was also noted with probes for types 16 and 18, and types 31, 33, and 35 in two cases from Australia and one from Papua New Guinea. It is concluded that as similar staining patterns and intensities occurred in cases from both areas, the aetiology is the same. The equivocal reactions noted in three cases were probably due to cross hybridisation rather than multiple infection.
Improved silver staining of nucleolar organiser regions in paraffin wax sections using an inverted incubation technique.
Authors: Coghill G , Grant A , Orrell J M , Jankowski J , Evans A T
Published: June 2007

A new simple modification to the silver staining of nucleolar organiser regions (AgNORs) was devised which, by performing the incubation with the slide inverted, results in minimal undesirable background staining, a persistent problem. Inverted incubation is facilitated by the use of a commercially available plastic coverplate. This technique has several additional advantages over other published staining protocols. In particular, the method is straightforward, fast, and maintains a high degree of contrast between the background and the AgNORs.