bluuebunny/crossref_metadata_2025 huggingface dataset:
Changes in cytomorphology of childhood lymphoblastic leukaemia at the time of disease relapse. Childhood Leukaemia Working Party of the United Kingdom Medical Research Council.
Authors: Lilleyman J S , Stevens R F , Hann I M , Gibson B E , Britton J A , Darbyshire P J , Oakhill A
Published: June 2007

AIMS--Children in a United Kingdom national trial for relapsed non-B lymphoblastic leukaemia (ALL) had their diagnostic and relapse marrow cytomorphology compared to see what changes occur during the evolution of the disease. METHODS--Each relapse slide was assessed blindly for French American British (FAB) type and other morphological features by a panel of three independent microscopists without reference to each other or any diagnostic material. Diagnostic slides had been assessed by the same panel on an earlier occasion. RESULTS--A total of 134 consecutive children was studied. Six (5%) were classified as FAB type L2 at diagnosis, compared with 18 (13%) at relapse (a difference of 9%). Twenty two (16%) changed their FAB type, 17 (13%) from L1 to L2 and five (4%) from L2 to L1. The FAB score fell at relapse in 34 children and rose in 14, a difference of 14%. Cell size was the commonest feature to change (increasing in 22 and diminishing in nine) followed by prominent nucleoli (appearing in 21 and disappearing in six). Forty four (33%) children had vacuolated blasts at diagnosis, compared with 48 (36%) at relapse. Twenty five changed their vacuole score substantially, 14 gaining > 10% and 11 falling < 10%. CONCLUSIONS--These findings reflect the variability of lymphoblast cytomorphology, but also show a trend for cells to have more prominent nucleoli and greater size at relapse. Factors controlling these features of the FAB type are unknown, but they may simply be related to the growth fraction of a particular disease and not to any lineage specific biological feature.
Pseudoangiosarcomatous carcinoma of the genitourinary tract.
Authors: Pitt M A , Morphopoulos G , Wells S , Bisset D L
Published: June 2007

Two cases of pseudoangiosarcomatous carcinoma of the genitourinary tract, arising in the vulva in one and the bladder in the other, are presented. In case 1, an 84 year old woman, the vulvectomy specimen contained an irregular ulcerated tumour, infiltrating the left labia and extending into the clitoris. In case 2, a 59 year old woman, the excised bladder showed diffuse thickening of its wall by infiltrating haemorrhagic tumour. Both tumours showed focal keratinisation. This, in association with the presence of atypical squamous epithelium, immunohistochemistry and ultrastructural analysis, led to a diagnosis of pseudosarcomatous carcinoma in both cases. Pseudoangiosarcomatous carcinoma should be considered in the differential diagnosis of malignant angiomatoid tumours, particularly those that arise at sites, like the genitourinary tract, where angiosarcoma is rare.
Detection of Chlamydia trachomatis antigen by enzyme immunoassay: importance of confirmatory testing.
Authors: Fonseca K , Megran D W , Anand C M
Published: June 2007

AIM--To determine when a fluorescence assay for Chlamydia trachomatis elementary bodies in the specimen buffer is of most value as a verification test for genital specimens reactive on screening enzyme immunoassay (EIA). METHOD--Genital swabs from high and medium prevalence populations were tested using EIA. Samples with absorbance values greater than the positive threshold and those within the range of 30% below this value were verified by the MicroTrak direct fluorescence assay (DFA) test. Quotients derived from the threshold value and specimen absorbances were used to establish confidence limits for the EIA. RESULTS--Of 13,283 swabs tested, 474 from the high risk group and 236 from the medium risk group were reactive on EIA and confirmed by DFA. Thirty six (5.9%) patients with confirmed reactive samples would have been missed if the kit criteria alone were followed. When confidence limits were applied to the calculated quotients, only those samples with an EIA quotient of > or = 4.0 were universally confirmed by the DFA. CONCLUSION--A scheme of testing which uses the DFA to verify EIA reactive specimens over a specified range was found to improve the sensitivity and specificity of the EIA screening test.
Iron metabolism and fungal infections in patients with haematological malignancies.
Authors: Iglesias-Osma C , Gonzalez-Villaron L , San Miguel J F , Caballero M D , Vazquez L , de Castro S
Published: June 2007

AIM--To determine whether iron metabolism influences the incidence of systemic fungal infection in patients with haematological malignancies. METHODS--The study population comprised 74 patients who had undergone myeloablative chemotherapy. Systemic fungal infections were classified as confirmed (histological confirmation or characteristic septate hyphae) or possible (antibiotic resistant fever which resolved following administration of intravenous amphotericin B, together with either typical radiographic lesions or massive oropharyngeal candidiasis). Parameters of iron metabolism included serum iron concentrations, total iron binding capacity, serum transferrin, and ferritin concentrations and transferrin saturation values. RESULTS--Patients who developed a fungal infection had substantially increased transferrin saturation values and ferritin concentrations at diagnosis together with low serum transferrin and high serum iron concentrations. This profile was present in patients with a fungal infection regardless of the underlying haematological disorder. CONCLUSION--Increased transferrin saturation values and high ferritin concentrations may be additional risk factors for the development of systemic fungal infection in patients with haematological malignancies.
p53 expression in hepatocellular carcinoma in a population in Singapore with endemic hepatitis B virus infection.
Authors: Wee A , Teh M , Raju G C
Published: June 2007

AIMS--To study the expression and clinical significance (if any) of p53 protein in hepatocellular carcinomas (HCC) arising in a population with endemic hepatitis B virus (HBV) infection. METHODS--Immunohistochemical staining was performed on formalin fixed, paraffin was embedded histological sections of 46 HCC cases using an antihuman p53 monoclonal antibody; serial sections were also stained for hepatitis B surface antigen (HBsAg), hepatitis B core antigen (HBcAg) and alpha fetoprotein (AFP). Nuclear p53 staining was assessed according to intensity (absent, weak or strong) and extent (< 5%, 6-25%, 26-50%, and > 50%) of positive cells. Tissue HBsAg, HBcAg and AFP were recorded as absent or present. RESULTS--The p53 protein was expressed in 35% (16 of 46) of HCCs; the positive rate in grade III/IV tumours (13 of 31; 42%) was higher than in grade I/II tumours (three of 15; 20%) but this was not statistically significant. HBsAg positive tumours showed almost the same proportion of p53 staining (11 of 29; 38%) as HBsAg negative ones (five of 17; 29%). CONCLUSIONS--The p53 protein was expressed in 35% of HCC cases. There was no statistically significant correlation between HBV infection and p53 protein expression. Similarly, there was no definite correlation between p53 positivity and tumour size, histological grade or vascular invasion.
Dieulafoy's disease associated with early gastric cancer.
Authors: Leone O , Zanelli M , Santini D , Minni F , Marrano D
Published: June 2007

In the past different terms have been used to define the vascular malformations of Dieulafoy's disease--for example, calibre persistent artery of the stomach, cirsoid aneurysm and gastric atherosclerosis. A case of Dieulafoy's disease is described in a 41 year old man, who presented with symptoms of anaemia and melaena, with particular attention paid to the morphological characterisation of the vascular histological lesions. Intimal hyperplasia with a non-concentric proliferation of myointimal cells, areas of muscular degeneration, aspects of vascular neoformation of the arterial wall, and other findings are reported. An association between an early diffuse adenocarcinoma and parietal anomalies of Dieulafoy's disease is illustrated.
Degenerative changes in myometrium simulating diffuse leiomyomatosis after treatment with gonadotrophin releasing hormone analogue.
Authors: Jeffers M , Cowan C , Lunan C B
Published: June 2007

Degenerative changes are encountered relatively frequently in uterine leiomyomas. Morphologic changes within leiomyomas, particularly necrosis and alterations in cellularity, have been described following treatment with gonadotrophin releasing hormone analogue, but the effects of this form of treatment on the morphology of the normal myometrium are less well documented. A case is reported of a 42 year old woman with a history of menorrhagia in whom a combination of degenerative and iatrogenic changes resulted in a histological appearance resembling diffuse leiomyomatosis.
Circulating endothelin-1 concentrations in patients with chronic hypoxia.
Authors: Ferri C , Bellini C , De Angelis C , De Siati L , Perrone A , Properzi G , Santucci A
Published: June 2007

AIMS--To evaluate the behavior of plasma endothelin-1 in patients with chronic hypoxia. METHODS--Fifteen male patients (mean age 52.1 +/- 3.1 years) with mild chronic obstructive pulmonary disease (COPD) were studied. Twelve healthy men (mean age 48.3 +/- 5.4 years) served as controls. Both patients and controls underwent standard pulmonary function tests, echocardiographic evaluation, and arterial blood gas evaluation. Blood samples for endothelin-1 assay were taken from a previously incannulated antecubital vein after 60 minutes of rest in the supine position. Endothelin-1 was measured by radioimmunoassay after extraction from plasma. RESULTS--Patients with chronic hypoxia had lower PaO2 values (66.1 +/- 6.2 mmHg) than controls (83.8 +/- 2.7 mmHg) but PaCO2 values were similar (38.1 +/- 2.5 v 36.7 +/- 3.1 mmHg, respectively). Arterial pulmonary pressure, therefore, was higher in patients (18.1 +/- 3.7 mmHg) than in controls (10.4 +/- 2.7 mmHg) as were circulating endothelin-1 concentrations (1.22 +/- 0.36 v 0.57 +/- 0.1 pg/ml). Furthermore, plasma endothelin-1 concentrations were negatively correlated with PaO2 and directly correlated with pulmonary pressure levels. No significant correlations were found in controls. CONCLUSIONS--These results show a clear relation between chronic hypoxia and circulating endothelin-1 concentrations. Therefore, chronic hypoxia may be regarded as an important stimulus for endothelin-1 release and as one of the main contributors to increased vasoconstriction in the vascular pulmonary bed which often accompanies lung disease.
Activated phenotype in neutrophils and monocytes from patients with primary proliferative polycythaemia.
Authors: Westwood N B , Copson E R , Page L A , Mire-Sluis A R , Brown K A , Pearson T C
Published: June 2007

AIM--To investigate whether monocytes and neutrophils from patients with primary proliferative polycythaemia (PPP) exhibit increased expression of markers of cell activation and, if so, whether they are associated with the phagocytic activity of these cells and concentrations of circulating cytokines. METHODS--Expression of CD11b, CD14, CD18, and CD64 on monocytes and neutrophils was assessed by flow cytometry. Phagocytosis was analysed using immunoglobulin opsonised Escherichia coli. Serum concentrations of granulocyte colony stimulating factor (G-CSF), granulocyte-macrophage CSF (GM-CSF) and macrophage CSF (M-CSF) were determined by bioassays, and interferon-gamma (IFN-gamma) by enzyme linked immunosorbent assay (ELISA). RESULTS--Patients with PPP (n = 18), when compared with normal subjects (n = 10), had increased percentages of CD64+ monocytes (52% v 36%) and neutrophils (42% v 11%) and of CD14+ neutrophils (36% v 18%). Monocytes from patients with PPP exhibited increased expression of CD64 (47 v 26) and of CD11b (65 v 36). These abnormalities were not found in patients with secondary (n = 8) or apparent (n = 13) polycythaemia. The percentage of neutrophils undergoing phagocytosis was higher in patients with PPP (mean 64%; n = 6) than in normal subjects (mean 42%; n = 5). G-CSF, GM-CSF and IFN-gamma concentrations in patients' serum samples were comparable with normal; M-CSF was not detected in any of the samples. There was no correlation between cytokine concentrations and the expression of CD11b, CD14, CD18, and CD64 on patients' phagocytes. CONCLUSIONS--Increased expression of CD11b and CD64 by monocytes, increased percentages of CD14+ and CD64+ neutrophils and the high phagocytic activity of neutrophils suggests that these cells are activated in vivo in patients with PPP. The phenotypic changes of PPP phagocytes were not associated with increased concentrations of circulating cytokines and probably reflect intrinsic abnormalities within the neoplastic PPP clone.
p53 protein accumulation in oesophageal squamous cell carcinomas and precancerous lesions.
Authors: Volant A , Nousbaum J B , Giroux M A , Roué-Quintin I , Metges J P , Férec C , Gouérou H , Robaszkiewicz M
Published: June 2007

AIMS--To investigate the immunohistochemical expression of p53 protein in oesophageal squamous cell carcinomas and in dysplastic areas of the oesophageal mucosa surrounding the tumours. METHODS--Biopsy samples were obtained from 20 patients with an oesophageal squamous cell carcinoma. Blocks of the tumours and of the surrounding mucosa were immunostained with the monoclonal antibody DO-7. RESULTS--Fourteen of the 20 carcinomas were positive for p53 (70%). The frequency of p53 overexpression increased with the differentiation of the tumour. Nine out of 13 dysplastic specimens were positive for p53 (69%): eight cases with severe dysplasia and one case with moderate dysplasia. No p53 immunostaining was detected in normal oesophageal epithelium. All p53 positive dysplastic specimens were taken from the mucosa adjacent to tumours that were also immunostained. In moderate dysplastic mucosa the p53 positive cells were located in the proliferative basal zone, whereas in severe dysplasia the immunostained cells increased in number and spread to upper cell layers of the epithelium. CONCLUSION--This study supports the hypothesis that TP53 gene is frequently involved in the development of oesophageal squamous cell carcinoma and that p53 protein accumulation is an early event in human oesophageal carcinogenesis.
Expression of HLA-ABC, HLA-DR and intercellular adhesion molecule-1 in oesophageal carcinoma.
Authors: Rockett J C , Darnton S J , Crocker J , Matthews H R , Morris A G
Published: June 2007

AIM--To examine the expression of HLA-ABC and HLA-DR major histocompatibility (MHC) antigens and intercellular adhesion molecule (ICAM)-1 in normal, inflamed, metaplastic, and neoplastic oesophageal tissue and in freshly disaggregated tumours. METHODS--Sequential sections of frozen tissue and cytospins of freshly disaggregated tumour were stained using the ABC peroxidase system and monoclonal antibodies specific for HLA-ABC, HLA-DR and ICAM-1. RESULTS--Normal oesophageal tissue showed positive staining for HLA-ABC in the basal layers of the oesophageal squamous epithelium and on the epithelial cells of the submucosal oesophageal glands. HLA-DR and ICAM-1 were not detected in either of these cell types. In 20 of 37 (54%) carcinomas HLA-ABC was expressed weakly, with heterogeneous expression in nine (24%). Two tumours showed strong expression of HLA-ABC, but 15 of 37 (41%) were negative. HLA-DR and ICAM-1 were expressed weakly in six of 37 (16%) carcinomas without correlation with each other or with the expression of HLA-ABC. CONCLUSIONS--HLA-ABC is absent from a high proportion of oesophageal carcinomas (41%) and is otherwise variably and weakly expressed with strong expression in only a small fraction (3%). In other carcinomas there is a higher level of HLA-ABC expression. This discrepancy may partly explain the aggressive nature of oesophageal carcinomas. HLA-DR and ICAM-1 are not normally expressed on those cells from which oesophageal carcinomas are thought to arise. The limited expression found here could suggest a partial or inhibited immune response against oesophageal carcinoma. In vivo repressive factors may be involved.
Commercially available versus home produced egg media for isolation of mycobacteria.
Authors: Harris G , Rayner A , Watt B
Published: June 2007

AIMS--To compare the growth of clinical isolates of mycobacteria on in-house and commercial egg media. METHODS--Fresh test media were inoculated with dilutions of the test organisms and growth, colonial morphology and microscopic appearance were compared blindly by two observers. The process was repeated after the test media had been stored for three months. The user friendliness of each of the test media was also assessed. RESULTS--There was no difference in the microscopic appearance of any given mycobacterial species between different media. All of the test media grew the test species, although Mycobacterium bovis required four weeks on BioMerieux media, compared with two weeks on the other media. There was little obvious effect of storage on any of the media, except with M kansasii. Individual species gave characteristic colonial appearances on inhouse media; all of the commercial media gave non-specific colonial appearances that made presumptive identification very difficult. There were clear differences in the user friendliness of different media. CONCLUSIONS--Although commercially available egg media in general gave good growth of mycobacteria, only in-house media combined good growth with useful colonial features.
Histological features of the thyroid gland in a patient with lithium induced thyrotoxicosis.
Authors: Mizukami Y , Michigishi T , Nonomura A , Nakamura S , Noguchi M , Takazakura E
Published: June 2007

A 26 year old woman with lithium induced thyrotoxicosis is reported. The thyrotoxicosis was associated with a non-tender diffuse goitre and a low radioiodine uptake by the gland. The thyrotoxicosis was reversible and remitted on withdrawal of the drug. The histopathological alterations of the thyroid glad were characterised by extensive follicular cell disruption with no lymphocytic infiltration. It is postulated that lithium might directly damage thyroid follicular cells and that subsequent release of thyroglobulin into the circulation might be a cause of transient thyrotoxicosis.
Immunohistochemical staining of plastic embedded bone marrow trephine biopsy specimens after microwave heating.
Authors: McCluggage W G , Roddy S , Whiteside C , Burton J , McBride H , Maxwell P , Bharucha H
Published: June 2007

AIMS--To investigate (1) whether adequate immunohistochemical staining can be achieved on sections cut from plastic embedded bone marrow trephine biopsy specimens after microwave heating in citrate buffer; and (2) whether this immunohistochemical staining is comparable with that achieved on routine sections cut from paraffin wax embedded trephine biopsy specimens after decalcification procedures. METHODS--Sixty five consecutive bone marrow trephine biopsy specimens of more than 1 cm in length were divided transversely into two equal parts. One part was processed in paraffin wax followed by decalcification. The other part was embedded in the epoxyresin Polarbed 812 followed by the cutting of 1 micron sections. Both parts underwent immunohistochemical staining by an identical panel of antibodies. With Polarbed 812 plastic embedded sections, microwave heating in citrate buffer was undertaken before the application of antisera. RESULTS--On sections cut from plastic embedded material, immunohistochemical staining was generally satisfactory, easy to interpret and comparable with that achieved with paraffin wax embedded material. Exceptions were antibodies to neutrophil elastase and CD61 where immunostaining was consistently negative on plastic embedded sections. Immunohistochemical staining for CD20 was consistently more reliable on plastic embedded sections. CONCLUSIONS--The results provide evidence that, with few exceptions, satisfactory immunohistochemical staining is possible on plastic embedded bone marrow trephine biopsy specimens after microwave heating in citrate buffer. This, combined with the advantage of superior cellular morphology with semi-thin (1 micron) sections of plastic embedded material, make such embedding procedures the preferred method for the processing of bone marrow trephine biopsy specimens.
Down-regulation of Epstein-Barr virus nuclear antigen 1 in Reed-Sternberg cells of Hodgkin's disease.
Authors: Khan G , Naase M A
Published: June 2007

AIMS--To demonstrate Epstein-Barr virus (EBV) encoded nuclear antigen 1 (EBNA-1) gene expression in EBV associated disorders using a new monoclonal antibody (1H4-1) on routinely processed tissues. METHODS--The pressure cooker antigen retrieval method was used for the immunohistochemical demonstration of EBNA-1 gene expression in formalin fixed, EBV positive tissues from Hodgkin's disease, infectious mononucleosis, HIV associated non-Hodgkin's lymphomas, post-transplant lymphomas, and undifferentiated nasopharyngeal carcinoma (NPC). EBV encoded EBNA-2, latent membrane protein 1 (LMP-1) and BZLF-1 gene expression was also examined using commercially available monoclonal antibodies. RESULTS--Of the 34 EBER in situ hybridisation positive cases of Hodgkin's disease examined, none expressed EBNA-1 in the Reed-Sternberg cells. These cells were nevertheless strongly LMP-1 positive in all cases. Strong EBNA-1 staining was seen in all cases of EBER positive HIV associated non-Hodgkin's lymphoma (five of five), nasopharyngeal carcinoma (five of five), infectious mononucleosis (three of three), and post-transplant lymphoma (one of one). These cases also expressed LMP-1, EBNA-2 and BZLF-1, but at differing levels. CONCLUSION--The pressure cooker antigen retrieval procedure is a sensitive and reliable adjunct to immunohistochemistry, especially with antibodies which are otherwise ineffective on routinely processed tissues. The EBNA-1 gene is not expressed at detectable levels in the malignant cells of Hodgkin's disease, but is consistently expressed in other EBV associated disorders. This finding has important implications for the role of EBNA-1 in the biology of EBV.
Immunohistochemical analysis of T cell proliferation in normal tonsil and B cell lymphoma.
Authors: Wang J , Isaacson P G , Spencer J
Published: June 2007

A double immunohistochemical technique, incorporating MIB1 and CD3, was used to identify proliferating T cells in paraffin wax sections of normal tonsil and B cell lymphomas. The number of double stained T cells as a percentage of the total T cells was then determined. In normal tonsil and follicular lymphoma the follicle centre and T cell zones were counted independently. In normal tonsil very few T cells in the follicle centre expressed MIB1. Proliferating T cells were concentrated in the T cell zones. The same pattern was observed in follicular lymphoma. In contrast, the percentage of T cells expressing MIB1 was higher in mucosa associated lymphoid tissue type lymphoma and lymphocytic lymphoma, suggesting that T cell activation occurs in these tumours. The highest percentage of MIB1 positive T cells was observed in high grade lymphoma. This suggests that transformation to high grade lymphoma is associated with an increase in T cell activation.
Inverse relation of serum Helicobacter pylori antibody titres and extent of intestinal metaplasia.
Authors: Osawa H , Inoue F , Yoshida Y
Published: June 2007

AIMS: To clarify the relation between the serum titre of anti-Helicobacter pylori (H pylori) antibody and the extent of intestinal metaplasia of the gastric mucosa. METHODS: The serum anti-H pylori IgG titres of 95 asymptomatic individuals (mean age 65 years) undergoing an annual health examination were measured and compared with the extent of intestinal metaplasia (absent, moderate, or extensive), determined by examination of multiple endoscopic mucosal biopsy specimens. Serum pepsinogen I (PGI) levels, as a marker for gastric atrophy, were also measured. RESULTS: The prevalence of seropositivity for H pylori antibody was high (> 80%), regardless of the extent of metaplasia. However, there was a negative association between the extent of metaplasia and the anti-H pylori titre: 75% of the subjects in the group without metaplasia had high (3+) antibody levels, as did 43% with moderate, and 37% with extensive metaplasia (absent v extensive). The inverse relation between the titre and the extent of metaplasia was evident when examined in those with normal PGI (> 30 ng/ml), whereas no such relation was apparent in subjects with low PGI (< or = 30 ng/ml). CONCLUSIONS: The anti-H pylori titre correlates inversely with the extent of intestinal metaplasia, particularly in subjects with less marked gastric atrophy.
Lipoproteins in pregnant women before and during delivery: influence on neonatal haemorheology.
Authors: Uberos-Fernández J , Muñoz-Hoyos A , Molina-Carballo A , Puertas-Prieto A , Valenzuela-Ruiz A , Ruiz-Cosano C , Molina-Font J A
Published: June 2007

AIMS: To investigate whether the lipid profile of pregnant women during parturition differs from the profile at previous stages of pregnancy and to determine the effects of maternal lipid changes on fetal or neonatal haemorheology. METHODS: Sixty pregnant women were studied, divided into two groups. Group 1 contained 30 women of mean age of 27 (SD 3) years and gestational age > 38 weeks in whom delivery had not yet begun; all these pregnancies followed an uncomplicated course and there was no evidence of any fetal pathology from previous obstetric examinations. All the women reached term and birth weight was 3340 (350) g. Group 2 contained women of mean age 26 (4) years, in whom delivery was ongoing, all of whose pregnancies reached term. The following variables were determined in all cases: total cholesterol, triglycerides, high density lipoproteins (HDL), low density lipoproteins (LDL), free fatty acids and phospholipids, and apoprotein A (apo-A) and apoprotein B (apo-B). Serum and plasma viscosity was measured with a capillary viscosimeter. RESULTS: The apo-B/apo-A and HDL/apo-A ratios increased during delivery, indicating that in pregnant women these atherogenic indices are raised during delivery compared with previous gestational stages. Significant correlation coefficients were obtained between maternal lipids (triglycerides, total cholesterol, LDL, total cholesterol/HDL, and LDL/HDL) and plasma viscosity in the neonate. CONCLUSIONS: Plasma atherogenic indices increase progressively until birth. These changes have implications for neonatal haemorheology because they cause an increase in plasma viscosity.
Observer variability in histopathological reporting of non-small cell lung carcinoma on bronchial biopsy specimens.
Authors: Burnett R A , Howatson S R , Lang S , Lee F D , Lessells A M , McLaren K M , Nairn E R , Ogston S , Robertson A J , Simpson J G , Smith G D , Tavadia H B , Walker F
Published: June 2007

AIMS: To evaluate the ability of histopathologists to sub-classify non-small cell lung carcinomas on bronchial biopsy material using the current World Health Organisation (WHO) classification. METHODS: Twelve histopathologists each reviewed 100 randomly selected bronchial biopsy specimens which had originally been reported as showing non-small cell lung carcinoma. For each case, two sections were circulated, one stained by haematoxylin and eosin and the other by a standard method for mucin (alcian blue/periodic acid Schiff). The participants were allowed to indicate their degree of confidence in their classification of each case. A standard proforma was completed and the results were analysed using kappa statistics. RESULTS: Where the participants were confident in their classification, they were actually quite good at sub-classifying the non-small cell carcinoma sections (kappa = 0.71, standard error = 0.058). Overall, however, the results were only fair (kappa = 0.39, standard error = 0.034). CONCLUSIONS: The majority of non-small cell lung carcinomas can be correctly categorised on adequate bronchial biopsy material. Where a confident diagnosis was made, both squamous carcinoma (kappa = 0.73) and adenocarcinoma (kappa = 0.83) were well recognised.
Expression of TIA-1 and TIA-2 in T cell malignancies and T cell lymphocytosis.
Authors: Matutes E , Coelho E , Aguado M J , Morilla R , Crawford A , Owusu-Ankomah K , Catovsky D
Published: June 2007

OBJECTIVE: To investigate the reactivity with TIA-1 and TIA-2, two monoclonal antibodies that recognise, respectively, granular structures in T lymphocytes and the T cell receptor chain in cells from a variety of T cell disorders. METHODS: Cytoplasmic staining with TIA-1 and TIA-2 was carried out by the immunoalkaline phosphatase anti-alkaline phosphatase technique in 67 cases with a T cell disorder: 31 large granular lymphocyte (LGL) leukaemia, nine T-prolymphocytic leukaemia (T-PLL), five Sezary syndrome, four peripheral T cell lymphoma (PTCL), 13 T cell lymphocytosis, and five T-acute lymphoblastic leukaemia (T-ALL). All had over 75% abnormal T cells which were CD2+, CD3+, CD5+, CD7+, and negative with B cell markers. RESULTS: TIA-1 was positive in 77% cases of LGL leukaemia and half of the PTCL and T-ALL, whereas it was negative in all Sezary syndrome and most T-PLL (8/9) and reactive T-lymphocytosis (10/13). In LGL leukaemia, TIA-1 was positive irrespective of the membrane phenotype, whether CD8+, CD4- or CD4+, CD8-, and was more often positive in cases where cells were CD16+, CD56+, or CD57+. TIA-2 was positive in 60% of cases encompassing all diagnostic types of T cell disorder. There was no correlation between TIA-2 expression and that of other T cell markers, activation antigens, and natural killer markers. CONCLUSIONS: The pattern of TIA-1 expression in T cell malignancies may help in the differential diagnosis among LGL leukaemia (high expression), T cell lymphocytosis and other T cell diseases (low expression). As TIA-2 is expressed in over 95% mature T lymphocytes and thymic cells, its assessment may be useful to demonstrate aberrant phenotypes which can be exploited for detecting minimal residual disease.
Sleeping position and upper airways bacterial flora: relevance to cot death.
Authors: Bell S , Crawley B A , Oppenheim B A , Drucker D B , Morris J A
Published: June 2007

The hypothesis that the prone sleeping position is associated with accumulation of upper airways secretions and increased bacterial growth was investigated in adults. Ten subjects with upper respiratory tract infection lay prone for one hour and then supine for one hour. Nasal swabs after the prone period yielded higher bacterial counts than swabs obtained after the supine period. This result could be relevant to sudden infant death syndrome (SIDS), as infants who sleep in the prone position are at increased risk of SIDS and one theory is that death is caused by toxins produced by bacterial overgrowth in the upper respiratory tract following a viral infection.
A rapid and highly specific technique to detect hepatitis C RNA in frozen sections of liver.
Authors: Sergi C , Goeser T , Otto G , Otto H F , Hofmann W J
Published: June 2007

AIMS: To develop a polymerase chain reaction (PCR) based technique that would permit the rapid and highly specific detection of hepatitis C virus (HCV) RNA extracted from frozen liver tissue. METHODS: Samples of liver tissue from 18 patients undergoing orthotopic liver transplantation were studied. Nine patients were HCV positive. Total RNA was extracted from between one and 10 sections, 10 microns thick, from each tissue sample. HCV RNA was amplified by (1) conventional, multistep reverse transcription PCR (RT-PCR) and by (2) combined, single step RT-PCR using coupled oligonucleotide primers based on the sequence of the 5' untranslated region of the viral genome. Positive results were confirmed by dot blot analysis using a digoxigenin labelled oligoprobe (Alx 89). RESULTS: HCV RNA was detected in the nine HCV positive patients by both conventional and combined RT-PCR. HCV RNA was not detected in the HCV negative patients. As little as 500 ng total RNA was needed as the template to yield detectable amounts of amplified cDNA. The digoxigenin labelled oligoprobe hybridised with HCV RNA positive specimens only. CONCLUSIONS: The combined, single step RT-PCR is a rapid and sensitive technique for detecting HCV RNA in frozen liver tissue.
Histology of chronic gastritis with and without duodenitis in patients with Helicobacter pylori infection.
Authors: Phull P S , Price A B , Stephens J , Rathbone B J , Jacyna M R
Published: June 2007

AIM: To compare the histological characteristics of Helicobacter pylori positive chronic gastritis in patients with and without associated duodenitis. METHODS: Gastric mucosal biopsy specimens were obtained from patients undergoing endoscopy for dyspepsia. Severity of gastritis and density of H pylori infection were graded according to the Sydney system. RESULTS: Of the 69 patients studied, 15 had normal histology, 22 had chronic gastritis only (77.3% H pylori positive), 21 had duodenitis (90.5% H pylori positive), and 11 had other diagnoses. In the H pylori positive patients, the median gastritis score was higher in the duodenitis group (6, range 3-9) than in the chronic gastritis only group (5, range 2-8), because of greater neutrophil activity scores in patients with duodenitis (median score 2 v 1). There were no differences in the density of H pylori infection, inflammation, atrophy, or intestinal metaplasia between patients with chronic gastritis only and those with duodenitis. CONCLUSIONS: These results suggest that H pylori positive patients with duodenitis have a more severe form of gastritis than those without associated duodenal inflammation. This is because of increased neutrophil activity, which seems to be independent of the density of H pylori infection.
Immunohistochemical analysis of bcl-2 expression in transitional cell carcinoma of the bladder.
Authors: Shiina H , Igawa M , Urakami S , Honda S , Shirakawa H , Ishibe T
Published: June 2007

AIMS: To evaluate the expression of bcl-2 in transitional cell carcinoma (TCC) of the bladder; to compare bcl-2 expression with clinicopathological findings, p53 immunoreactivity, proliferating cell nuclear antigen (PCNA) expression, 2c deviation index (2cDI), 5c exceeding rate (5cER), and the mean nuclear area (MNA). METHODS: Cystectomy specimens from 77 patients with untreated, non-metastatic TCC of the bladder were studied. Expression of bcl-2, p53 and PCNA was detected immunohistochemically using the following monoclonal antibodies: bcl-2/124, DO-7 and PC10, respectively. Nuclear DNA content was analysed using static cytometry. RESULTS: Bcl-2 was expressed in 19 (24.7%) of 77 TCCs and in 74 (96.1%) of 77 normal samples of transitional epithelium (taken from normal tissue adjacent to the tumour in each case). In all cases, bcl-2 immunoreactivity was more intense in normal transitional epithelium than in TCC. In normal transitional epitehlium and superficial TCC bcl-2 immunoreactivity was observed at the basal layer, and not at the invasive front. Bcl-2 immunoreactivity was invesely correlated with histological grade and p53 immunoreactivity, and was not correlated with the pT category, disease progression, PCNA expression, 2cDI, 5cER, and the MNA. No significant correlation was found between bcl-2 expression and overall survival. CONCLUSIONS: Bcl-2 expression in TCC of the bladder seems to be associated with a less aggressive phenotype and does not play an important role in tumour progression.
Necropsy study of the association between sudden cardiac death, cardiac isoenzymes and contraction band necrosis.
Authors: Hopster D J , Milroy C M , Burns J , Roberts N B
Published: June 2007

AIMS: To assess whether a quantitative analysis of myocardial contraction bands could aid the postmortem identification of early myocardial infarction, especially if used in conjunction with cardiac isoenzyme activities. METHODS: Sixty four coroner's necropsies were grouped by gross and histological findings into 26 cases of definite non-cardiac death, 12 cases of definite myocardial infarction and 26 cases in which there was occlusive coronary artery atheroma, but no demonstrable evidence of infarction. Using multiple sections of left ventricular myocardium stained with Heidenhain's iron haematoxylin, the number of myocardial cells containing contraction bands per unit area was quantified. The results were analysed statistically using logistic regression, and were then compared and combined with results from the statistical analysis of postmortem cardiac isoenzymes that had recently been undertaken on the same cases. RESULTS: The number of cells containing contraction bands per unit area was higher in cases of definite myocardial infarction compared with those of non-cardiac deaths. In addition, cases of occlusive coronary artery atheroma only could be identified, indicating the presence of early myocardial infarction. The accuracy of this identification could be improved by combining these results with the results from the statistical analysis of postmortem cardiac isoenzymes. CONCLUSION: The quantitative assessment of myocardial contraction band necrosis can provide useful additional information in cases of sudden death, where myocardial infarction is suspected but not identified on routine histological staining. The value of the information obtained is increased when used in conjunction with the postmortem measurement of cardiac isoenzyme activities.
Multicentre evaluation of the Thrombotest International Sensitivity Index used with a steel ball coagulometer.
Authors: van den Besselaar A M , Peters R H
Published: June 2007

AIM: To compare the International Sensitivity Index (ISI) of the Thrombotest reagent used with a steel ball coagulometer (KC) to the ISI of the same reagent used with the manual (tilt tube) technique. METHODS: The study was carried out by eight laboratories using their own KC instrument and method of testing. All laboratories used the same batch of Thrombotest to determine the clotting times of fresh blood samples from 20 local healthy volunteers and 60 patients on long term oral anticoagulant therapy. KC clotting times were plotted against manual clotting times on double logarithmic scales. Orthogonal regression lines were calculated to assess the ISI. RESULTS: In two laboratories the ISI of the KC method was lower than that of the manual method; these differences, however, were 2% or less. In the other laboratories no clinically important differences were observed between ISI values obtained. However, the clotting times determined with the KC methods were shorter than the manual values. CONCLUSIONS: The ISI of Thrombotest determined with the KC methods was very similar to the manual value. Therefore, use of the ISI value supplied by the manufacturer without adjustment is justified. The mean normal prothrombin time, however, must be determined locally.
Quantification of HIV DNA in the brain by PCR: differences between fresh frozen and formalin fixed tissue.
Authors: Davison F , An S F , Scaravilli F
Published: June 2007

HIV-1 DNA extracted from frozen and formalin fixed brain tissue can be detected using PCR. This work has been extended by amplifying, using semiquantitative PCR, HIV DNA extracted from frontal lobe tissue of 16 patients with AIDS (eight positive and eight negative for p24 antigen). DNA was amplified using HIV-1 pol gene digoxigenin labelled primers and detected by chemiluminescence and densitometry. Cloned standards were amplified in parallel for quantification. HIV DNA levels detected in frozen tissue showed a correlation with p24 positivity and the severity of the histological diagnosis. This correlation was less clear in the formalin fixed material.
Topoisomerase II alpha mRNA and tumour cell proliferation in non-Hodgkin's lymphoma.
Authors: Lohri A , Reuter J , Gudat F , Herrmann R
Published: June 2007

AIMS: To elucidate potential mechanisms of drug resistance, levels of topoisomerase II alpha mRNA, a target for cytostatic drugs, were measured in cryopreserved tumour tissue from 36 patients with non-Hodgkin's lymphoma. To evaluate the potential association between topoisomerase II alpha and cell proliferation, Ki-67 immunostaining was also assessed. METHODS: The study population comprised 13 patients with low grade and 20 with high grade non-Hodgkin's lymphoma. Three patients had recurrent disease. Topoisomerase II alpha mRNA was quantitated by using reverse transcription polymerase chain reaction (RT-PCR) and the PCR product measured by using HPLC. The MIB-1 monoclonal antibody was used for Ki-67 immunostaining. RESULTS: Levels of topoisomerase II alpha mRNA correlated strongly with the Ki-67 labelling index and were higher in high grade than in low grade lymphomas. Patients in complete clinical remission of high grade lymphoma had a higher Ki-67 labelling index and tended to have higher topoisomerase II alpha mRNA levels. CONCLUSIONS: Although topoisomerase II alpha mRNA levels may be indicative of sensitivity to drugs, it is more likely that they reflect the proliferation status of the cell, which in turn involves a large number of additional molecular systems that influence response to treatment.
Oestrogen and progesterone receptor assessment in core biopsy specimens of breast carcinoma.
Authors: Zidan A , Christie Brown J S , Peston D , Shousha S
Published: June 2007

AIMS: To assess the suitability of core biopsy specimens for the immunohistological assessment of oestrogen and progesterone receptors in breast carcinoma. METHODS: Thirty consecutive cases of clinically palpable breast carcinoma, from which both core and excision biopsy specimens were available, were examined. Routinely processed paraffin wax sections were stained using the specific monoclonal antibodies 1D5 (Dako) for oestrogen receptor and NCL-PGR (Novocastra) for progesterone receptor, after an antigen retrieval step using a pressure cooker. Staining results were assessed using the H score system with the results being expressed as negative, weakly positive, moderately positive or strongly positive. RESULTS: Twenty six biopsy specimens contained enough tumour tissue for assessment. Absolute agreement between scoring categories was seen in 19 (73%) cases for oestrogen receptors. However, when all positive categories were added together, agreement between core and excision biopsy specimens increased to 93% (24 cases). Disagreement was seen only in two cases which stained positive in the core biopsy specimens and negative in the excision biopsy specimens. For progesterone receptors, the absolute agreement between all scoring categories was seen only in 11 (42%) cases. When all positive categories were considered together, agreement increased to 69% (18 cases). Five cases were progesterone receptor positive in core but not in excisional biopsy specimens, while three cases were negative in core but positive in excisional biopsy specimens. CONCLUSIONS: The results suggest that core biopsy specimens can be reliably used for oestrogen receptor assessment, but are less reliable for progesterone receptor assessment, probably because of a greater heterogeneity of progesterone receptor staining.
Immunocytochemical expression of tissue specific transcription factor-1 in lung carcinoma.
Authors: Di Loreto C , Di Lauro V , Puglisi F , Damante G , Fabbro D , Beltrami C A
Published: June 2007

AIMS: To investigate the immunocytochemical expression of the tissue specific transcription factor-1 (TTF-1) on cytological specimens of small cell lung carcinoma (SCLC) and to establish its value in the cytological diagnosis of lung cancer. METHODS: For each case, the diagnosis was made on cytological specimens and confirmed on subsequent bronchial biopsy specimens. TTF-1 was detected immunocytochemically using the avidinbiotin complex technique with a rabbit antiserum. Expression of TTF-1 was evaluated in 41 cases of SCLC and 17 cases of non-small cell carcinoma (NSCC). The latter were subdivided into eight cases of adenocarcinomas and nine cases of squamous cell carcinomas (SCC). RESULTS: Positive nuclear immunoreactivity to TTF-1 was identified in 38 (92.7%) of the 41 cases of SCLC, in five (62.5%) of eight cases of adenocarcinoma, and one (11%) of nine cases of SCC. A significant difference was observed between the two main groups, SCLC and NSCC. A comparison between SCLC and adenocarcinoma and SCC showed that TTF-1 expression was significantly different. TTF-1 immunoreactivity was not detected in the inflammatory cells of the same cases. CONCLUSIONS: TTF-1 is strictly associated with SCLC; it was weakly expressed in the various subtypes of NSCC. Although TTF-1 is not specific for SCLC, it can be used to highlight neoplastic cells to good effect when a large inflammatory component is present, and to differentiate SCLC from lymphoid infiltrates.
Prognostic value of bcl-2 expression in patients with operable carcinoma of the uterine cervix.
Authors: Tjalma W , Weyler J , Goovaerts G , De Pooter C , Van Marck E , van Dam P
Published: June 2007

AIM: To evaluate the patterns of bcl-2 expression in early stage cervical carcinoma; to compare bcl-2 expression with clinicopathological findings; and to assess its prognostic value. METHODS: Wertheim radical hysterectomy specimens from 76 patients (FIGO stages Ia-IIb) with untreated nonmetastatic invasive cervical carcinoma were studied. Expression of bcl-2 was detected immunohistochemically using a monoclonal antibody. A tumour was regarded as positive when more than 5% of the neoplastic cells exhibited bcl-2 immunoreactivity. RESULTS: Forty eight (63%) cervical carcinomas were scored as bcl-2 positive and 28 (37%) as bcl-2 negative. Most tumours showed heterogeneous cytoplasmic staining. Bc1-2 immunoreactivity did not correlate with tumour histology, tumour stage, presence of lymph node metastases, or involvement of the lymphovascular space. The five year survival rate for patients with bc1-2 negative tumours was 34% and was 71% for patients with bc1-2 positive tumours. On multiple regression analysis (Cox proportional hazards model), bc1-2 expression and vascular permeation were independent predictors of overall survival. CONCLUSIONS: Bcl-2 expression seems to be associated with less aggressive behaviour in early stage cervical carcinoma. The transition to bcl-2 independence may play an important role in tumour progression.
Calculation of System International Sensitivity Index: how many calibrant plasmas are required?
Authors: Dufty J M , Craig S , Stevenson K J , Taberner D A
Published: June 2007

AIMS: To determine the minimum number of calibrant plasmas required to assign an accurate System International Sensitivity Index (ISI). METHODS: Eighteen lyophilised adsorbed and eight frozen coumarin plasmas were despatched to 57 participating laboratories. Each centre determined the prothrombin time of all of the plasmas using their routine system of coagulometer and thromboplastin reagent and provided their system mean normal prothrombin time (MNPT). For each laboratory, the slope of the line relating local system results, including MNPT, to calibrated values obtained with BCT 441, RBT 1303 or RBT 90 was derived by orthogonal regression. The number of adsorbed calibrant plasmas included in the calculation was reduced, two at a time, from 18 to two. The validity of the System ISI was tested using the frozen coumarin plasmas. RESULTS: All systems tested gave higher International Normalised Ratios (INR) than those obtained manually with the reference thromboplastins. All systems therefore required correction. No changes were seen in the System ISI as the number of calibrant plasmas was reduced from 18 to 12. Below 12, inconsistent results were obtained with some systems and were dependent on plasma selection. CONCLUSIONS: Although System ISI could be assigned using as few as two calibrant plasmas, to avoid imprecise calibration, a minimum of 12 calibrant plasmas is recommended.