bluuebunny/crossref_metadata_2025 huggingface dataset:
Human papillomavirus DNA in glandular lesions of the uterine cervix.
Authors: Samaratunga H , Cox N , Wright R G
Published: June 2007

AIMS--To assess the role of human papillomavirus in the pathogenesis of adenocarcinoma in situ, endocervical glandular dysplasia (a presumed precursor of adenocarcinoma) and endocervical glandular epithelial giant cell change. METHODS--Viral detection was carried out using an in situ hybridisation technique on paraffin wax sections. Biotinylated probes for human papillomavirus types 6/11, 16/18, 31/33/35 were used with a colorimetric detection system. RESULTS--Nine out of 21 (43%) cases of adenocarcinoma in situ contained human papillomavirus types 16/18, one of which was also positive for 31/33/35. Ten cases of glandular dysplasia and four cases of glandular epithelial multinucleation did not react with the probes used. CONCLUSIONS--These results indicate that while adenocarcinoma in situ is strongly associated with human papillomavirus infection, endocervical glandular dysplasia and glandular epithelial multinucleation are probably not associated with the virus.
Biochemical and muscle studies in patients with acute onset post-viral fatigue syndrome.
Authors: Preedy V R , Smith D G , Salisbury J R , Peters T J
Published: June 2007

AIMS--To investigate in detail various biochemical and pathophysiological indices of muscle pathology in acute onset post-viral fatigue syndrome (PVFS). METHODS--Twenty three patients with PVFS (of mean duration 4.6 years) were subjected to needle biopsy for histomorphometry and total RNA contents. Plasma analysis included serology and creatine kinase activities. Indices of whole body mass were also measured--namely, whole body potassium content and plasma carnosinase activities. RESULTS--About 80% of the patients had serology indicative of persistent enteroviral infection as determined by VP1 antigen assay. Only about 10% of that same group of patients had serological indications of current enterovirus infection by IgM assay; a separate subset of 10% showed antibody changes suggestive of reactivation of Epstein-Barr virus. Quantitative morphometric analysis of skeletal muscle fibres indicated that the quadriceps muscle was normal or displayed only minor abnormalities in 22 patients. The Quetelet's Index (body mass index) and whole-body potassium values (index of lean body mass) were not affected in PVFS. The mean plasma carnosinase and creatinine kinase activities were also generally normal in these patients. The mean muscle RNA composition--mg RNA/mg DNA--was significantly reduced in acute onset PVFS by about 15%. The protein:DNA ratio was not significantly affected. CONCLUSIONS--Patients with acute onset PVFS, therefore, lose muscle protein synthetic potential, but not muscle bulk. Histopathology is consistent with these observations. These perturbations may contribute to the apparent feature of perceived muscle weakness associated with the persistent viral infection in the muscle themselves.
Acute myelomonocytic leukaemia following atypical congenital rubella.
Authors: Kelly S J , Gibbs T , Cheetham C H
Published: June 2007

The child of a woman immunised against rubella presented at 5 months with developmental delay and recurrent infection; she was shown to have congenital rubella. At 15 months she developed acute myelomonocytic leukaemia (AMML). Rubella is difficult to diagnose after immunisation. AMML has not been previously described in association with congenital rubella, as far as is known.
Diagnosis of childhood BK virus cystitis by electron microscopy and PCR.
Authors: Saitoh K , Sugae N , Koike N , Akiyama Y , Iwamura Y , Kimura H
Published: June 2007

A case of BK virus cystitis in a 5 year old boy is reported. This patient, who was not immunocompromised, had had acute cystitis for two weeks. Many intracytoplasmic inclusions were observed in urinary sediment smears stained by the Papanicolaou method. Electron microscopic examination showed virus particles, presumed to be human polyomavirus, in the nuclei of the degenerated urothelial cells. A DNA sequence of the BK virus was detected in 200-300 urothelial cells in Papanicolaou stained smears by the polymerase chain reaction. BK virus is an unusual cause of symptomatic cystitis in a healthy child.
Effect of embolisation of meningioma on Ki-67 proliferation index.
Authors: Paulus W , Meixensberger J , Hofmann E , Roggendorf W
Published: June 2007

The proliferation indices (PIs) were determined in 33 meningiomas using two monoclonal antibodies to the Ki-67 antigen (Ki-67 and MIB1). PIs obtained using Ki-67 on frozen material were intermediate between the lowest and highest MIB1 PIs determined in paraffin wax sections. Preoperative embolisation did not influence Ki67 PI, but four of the 15 embolised tumours showed substantially increased MIB1 PIs around embolisation necroses. Proliferation can be reliably assessed in routinely processed meningioma tissues using MIB1; and increased perinecrotic PIs may occur in embolised meningiomas without being an indication of malignancy.
Rapid diagnosis of cytomegalovirus encephalitis in patients with AIDS using in situ hybridisation.
Authors: Musiani M , Zerbini M , Venturoli S , Gentilomi G , Borghi V , Pietrosemoli P , Pecorari M , La Placa M
Published: June 2007

AIMS--To evaluate the presence of cytomegalovirus (CMV) DNA in the cerebrospinal fluid of patients with AIDS and suspected viral encephalitis using an in situ hybridisation assay with digoxigenin labelled CMV DNA probes. METHODS--The presence of CMV DNA was evaluated in cerebrospinal fluid cells of 10 patients with AIDS using in situ hybridisation. The positivity of CMV DNA was confirmed by the presence of CMV induced antigens in the same specimens. The presence of CMV DNA and CMV induced antigens was also analysed in peripheral blood leucocytes. The time required to perform the in situ hybridisation assay was about eight hours. RESULTS--The in situ hybridisation assay was sensitive, specific, and provided good resolution. Six patients proved positive for the presence of CMV DNA in CSF cells and all six also proved positive for CMV DNA in blood leucocytes. Of the six CMV positive patients, five were treated with specific antiviral drugs: of these, one died during the treatment while four clinically recovered after one month of treatment. CONCLUSIONS--The in situ hybridisation assay using digoxigenin labelled CMV DNA probes can be used as a valuable diagnostic test for the detection of CMV DNA in the cerebrospinal fluid cells of patients with suspected CMV encephalitis and can therefore prompt adequate antiviral therapeutic intervention.
Type 1 procollagen as a marker of severity of scarring after sternotomy: effects of topical corticosteroids.
Authors: Riaz Y , Cook H T , Wangoo A , Glenville B , Shaw R J
Published: June 2007

AIMS--To determine whether the abundance of newly formed collagen in healing surgical wounds correlated with scar severity, and whether topical application of steroid cream reduced new collagen formation in patients who have undergone median sternotomy. METHODS--Thirty three patients six weeks after sternotomy, and 12 controls were studied. Scars were photographed, and biopsy specimens from scars at sites treated or untreated with topical corticosteroids (clobetasol proprionate 0.5%) were examined using immunohistochemical staining for type 1 procollagen (PCP 1) and transforming growth factor beta (TGF-beta), and in situ hybridisation for type 1 procollagen messenger RNA (mRNA). RESULTS--The degree of hypertrophy of the scar and the abundance of PCP 1 immunostaining were ranked independently, blind, and a correlation between these two variables was observed (r = 0.604, p < 0.001). The PCP 1 immunostaining was accompanied by a great abundance of PCP 1 mRNA and only a slight increase in TGF-beta immunostaining, when compared with normal skin or mature scars. Following the application of topical corticosteroids, for either 48 hours or twice daily for seven days, there was no reduction in PCP 1 immunostaining nor the abundance of PCP 1 mRNA. CONCLUSIONS--These data suggest that the extent of new collagen formation as assessed by PCP 1 immunohistochemistry may be a useful marker of the exuberance of the scarring process following sternotomy, and that topical corticosteroids are ineffective in reducing this component of the fibrotic response.
Pancreatic enzymes in the epithelium of intrahepatic large bile ducts and in hepatic bile in patients with extrahepatic bile duct obstruction.
Authors: Terada T , Morita T , Hoso M , Nakanuma Y
Published: June 2007

AIM--To determine whether pancreatic enzymes are present in hepatic bile and in intrahepatic bile duct epithelium. METHODS--The activity and proteins of pancreatic enzymes (pancreatic alpha-amylase, lipase, trypsin/trypsinogen) in hepatic bile were investigated using biochemical and western blot analyses in 25 patients with extrahepatic bile duct obstruction. Immunolocalization of enzyme proteins was evaluated by immunohistochemistry in 20 necropsy livers with extrahepatic bile duct obstruction. RESULTS--Western blot analysis showed proteins of pancreatic alpha-amylase, lipase, and trypsin in 19 of 25 (76%), 10 of 25 (40%), and 14 of 25 (56%) patients, respectively. Pancreatic alpha-amylase and lipase activities was present in every bile specimen. Radioimmunoassay showed that trypsin was present in every bile sample. Immunohistochemically, the immunoreactivity of the three enzymes was present in epithelia and in the lumina of intrahepatic large bile ducts, septal bile ducts, and peribiliary glands in all cases. CONCLUSIONS--These results strongly suggest that biliary epithelia of larger intrahepatic ducts produce pancreatic alpha-amylase, lipase, and trypsin, and that these enzymes are secreted into the lumina of intrahepatic bile ducts.
Timed sequential analysis of creatine kinase in the diagnosis of myocardial infarction in patients over 65 years of age.
Authors: Chattington P , Clarke D , Neithercut W D
Published: June 2007

AIM--To assess the value of timed sequential analysis of creatine kinase (CK) activity for the early diagnosis of acute myocardial infarction (AMI) in patients over 65 years of age. METHOD--Samples were collected on admission and eight to 12 hours later from 156 patients over 65 years of age. Routine cardiac enzyme activities were determined and serial electrocardiograms (ECGs) recorded. The predictive value of timed samples for CK activity, standard cardiac enzyme activities, and ECGs was compared with the final diagnosis on discharge. RESULTS--Forty one patients had a discharge diagnosis of AMI, 83 of angina pectoris, and the remaining 32 patients had other diagnoses. Electrocardiograms had a sensitivity of 55% and a specificity of 96%, giving a predictive value of 86% for a negative and 84% for a positive ECG. Standard cardiac enzymes had a predictive value of 99% for a negative result but only 68% for a positive result. The logarithm of the rate of change of CK activity had a predictive value of 97% for a negative result and 95% for a positive result. CONCLUSION--This study has shown that slope analysis of CK activity can be used for the early diagnosis of AMI in patients over 65 years of age, and that this was not affected by the presence of possible confounding diagnoses.
Lymph node hyalinisation in rheumatoid arthritis and systemic sclerosis.
Authors: McCluggage W G , Bharucha H
Published: June 2007

AIMS--To review the histological features of lymph nodes excised from seven patients with rheumatoid arthritis and one with systemic sclerosis. METHODS--Lymph nodes excised from seven patients with rheumatoid arthritis and one patient with systemic sclerosis over a 10 year period were examined using the stains haematoxylin and eosin, periodic acid Schiff (PAS), Masson-trichrome, and Congo red for amyloid. RESULTS--Of the seven nodes examined from the cases of rheumatoid arthritis, three showed definite reactive follicular hyperplasia with a prominence of plasma cells in the interfollicular areas, two showed subtotal replacement of the node by numerous sarcoid like granulomata, and one contained a large central area of necrosis with a surrounding palisade of histiocytes. In all six cases, focal areas of PAS positive eosinophilic hyaline material were present, which did not stain with Congo red. In some cases this hyaline material was focally calcified. In the seventh patient with rheumatoid arthritis the excised lymph node was almost totally replaced by similar PAS positive hyaline material which showed extensive areas of calcification. The lymph node removed from the patient with systemic sclerosis similarly showed almost total replacement by PAS positive hyaline material. CONCLUSION--In all cases the nodes contained PAS positive extracellular hyaline material to a greater or lesser degree. The lymph nodes from two of the patients with rheumatoid arthritis contained numerous sarcoid like granulomata, further indicating a possible association between sarcoidosis and rheumatoid arthritis. Pathologists and clinicians should include rheumatoid arthritis and systemic sclerosis in their differential diagnosis of lymph node hyalinisation of unknown aetiology.
Cellular distribution of human leucocyte adhesion molecule ICAM-3.
Authors: Cordell J L , Pulford K , Turley H , Jones M , Micklem K , Doussis I A , Tyler X , Mayne K , Gatter K C , Mason D Y
Published: June 2007

AIMS--To describe the distribution of the recently cloned human leucocyte adhesion molecule ICAM-3 in normal and neoplastic tissues and cell lines. METHODS--A panel of four monoclonal antibodies to ICAM-3 were used to stain cell lines and sections of human lymphoid tissues using the alkaline phosphatase-anti-alkaline phosphatase immunocytochemical method (APAAP). RESULTS--In peripheral blood ICAM-3 was detected on monocytes, granulocytes, and most lymphocytes. In sections of human lymphoid tissue the antigen was also found on most lymphocytes, but many of the proliferating B cells found in the germinal centres of secondary lymphoid follicles were ICAM-3 negative. ICAM-3 was also found on neoplastic white cells (in chronic lymphocytic leukaemia, hairy cell leukaemia, acute and chronic myeloid leukaemia, and multiple myeloma) with the exception of Reed-Sternberg cells in Hodgkin's disease, many of which were negative. ICAM-3 was consistently absent from cells and tissues of non-haemopoietic origin. Endothelium (which expresses ICAM-1) was negative for ICAM-3, with the exception of vessels in some neoplastic lymphoid samples which showed variable staining for ICAM-3. CONCLUSIONS--These findings suggest that ICAM-3 is essentially restricted to the haemopoietic system and is reciprocal in its expression to ICAM-1, in that it is present on resting cells and its level falls as a result of cell activation.
Monitoring the acute phase response to vaso-occlusive crisis in sickle cell disease.
Authors: Stuart J , Stone P C , Akinola N O , Gallimore J R , Pepys M B
Published: June 2007

AIMS--To identify suitable acute phase proteins as objective markers of tissue ischaemia during painful vaso-occlusive crises in sickle cell disease. METHODS--The prodromal and established phases of 14 vaso-occlusive crises were studied longitudinally in 10 patients with sickle cell anaemia. Automated solid phase enzyme immunoassays were used to measure the fast responding acute phase proteins C-reactive protein and serum amyloid A protein. Slower responding glycoproteins (fibrinogen, orosomucoid, sialic acid and concanavalin-A binding) were measured in parallel. RESULTS--C-reactive protein and serum amyloid A protein increased early in crisis, sometimes within the early (prodromal) phase. Crises that resolved within 24 hours in hospital showed a minor and transient rise compared with crises that required treatment for four days or more. In eight crises treated by patients at home the acute phase response ranged from minor to a level consistent with extensive tissue ischaemia. CONCLUSIONS--Sensitive enzyme immunoassays for C-reactive protein and serum amyloid A protein are of potential value for monitoring the onset of tissue ischaemia in sickle cell crisis and for confirming subsequent resolution.
Polymerase chain reaction for screening clinical isolates of corynebacteria for the production of diphtheria toxin.
Authors: Pallen M J , Hay A J , Puckey L H , Efstratiou A
Published: June 2007

AIMS--To assess the performance of the polymerase chain reaction (PCR) when used to screen rapidly large numbers of corynebacteria for toxin production; and to determine the incidence of false positive PCR results with non-toxigenic Corynebacterium diphtheriae isolates. METHODS--Eighty seven recent British isolates of corynebacteria were assayed by PCR. All isolates were assayed from both blood and tellurite agar within a five day period. Thirty three non-toxigenic isolates of C diphtheriae from six countries were also tested by PCR and by the Elek immunodiffusion assay. RESULTS--There was complete concordance between the results of PCR and traditional methods on the recent British isolates, with one exception: an Elek positive "C ulcerans" isolate, which was PCR positive from tellurite but not from blood agar. One of the thirty three (3%) non-toxigenic isolates of C diphtheriae was PCR positive. CONCLUSIONS--These results suggest that PCR compares favourably with traditional methods for the detection of toxigenic corynebacteria and that it represents a powerful new tool in the diagnosis of an old disease.
Peptide based enzyme immunoassays for detecting hepatitis C antibodies in sera of people at high risk.
Authors: Gabriel F G , Teo C G
Published: June 2007

AIM--To evaluate the performance of three newly introduced enzyme immunoassays (EIAs) for hepatitis C virus (HCV) antibodies, based on synthetic oligopeptides as antigens. METHODS--Referred serum samples (n = 173) from people representing groups at high risk of HCV infection were studied. An EIA based on second generation recombinant polypeptide antigens was used for comparison. EIA reactivities were validated by testing repeatedly reactive samples in two recombinant antigen based immunoblot assays. RESULTS--In samples from patients with liver dysfunction and those with bleeding disorders sensitivity of the three peptide based EIAs, manufactured by Innogenetics NV, Biokit SA, and United Biomedical Inc., were all 93%; specificity and efficiency were all greater than 95%. In samples from blood donors (previously tested as positive by the Ortho and Abbott Second Generation EIA) specificity, sensitivity, and efficiency were 95% or greater in all three peptide assays. Sensitivity, specificity, and efficiency of the recombinant antigen based Ortho Second Generation EIA were 100%, 89%, and 93%, respectively, in sera of patients with liver disease and those with bleeding disorders; and 100%, 43%, and 83%, respectively, in prescreened samples from blood donors. CONCLUSION--The peptide EIAs are more specific but less sensitive than the Ortho EIA. Peptide based EIAs should be useful in validating the specificity of Ortho EIA reactivities.
Quantitative audit of the content of histopathology reports.
Authors: Campbell F , Griffiths D F
Published: June 2007

An audit of histopathology reports presents the problem that the output is textual and difficult to quantify. This makes the definition of an adequate report subjective and susceptible to observer variation. A procedure has been developed which allows the quantitative analysis of reports and facilitates the development of local reporting guidelines. A topic is selected; the auditor then lists the possible details that may be included in the report and notes how many reports from a sample include each detail. The results are discussed at a departmental meeting with the aim of agreeing on reporting guidelines. At a later date another sample of reports can be analysed for compliance with the guidelines and compared with the previous reports. Problems with compliance can be discussed further and at the audit meeting the guidelines may be amended appropriately, thus completing the audit cycle. This method of audit has the advantage that the results are quantitative and that the group discussion and re-examination of the guidelines has educational value.
Electron microscopic findings of parallel tubular granules in a case of chronic neutrophilic leukaemia.
Authors: Takemori N , Hirai K , Onodera R , Saito N , Namiki M
Published: June 2007

A case of chronic neutrophilic leukaemia (CNL) is reported. The diagnosis was based on leucocytosis with mature neutrophils, a raised leucocyte alkaline phosphatase score, negative Philadelphia chromosome, and extensive infiltration of neutrophils in various differentiation stages into the bone marrow. When viewed by light microscopy, these neutrophils were almost normal in appearance, except for the presence of ring shaped nuclei and cytoplasmic vacuoles. Electron microscopy showed that neutrophilic promyelocytes and early myelocytes in the bone marrow frequently possessed unique rounded granules consisting of clustered parallel tubules, of 29-31 nm in diameter, and occasional amorphous electron dense material. These parallel tubules showed a hexagonal array; the granules were termed parallel tubular granules (PTGs). PTGs were positive for electron microscopic myeloperoxidase, and were seen exclusively in neutrophilic promyelocytes and early myelocytes. These findings suggest that PTGs represent certain primary granules peculiar to immature neutrophils. Their presence might also be helpful in diagnosing CNL.
Leiomyosarcoma of soft tissue and pulmonary metastasis, both with osteoclast-like giant cells.
Authors: Matthews T J , Fisher C
Published: June 2007

A case is described of leiomyosarcoma with osteoclast-like giant cells in soft tissues which metastasised to the lung. The secondary tumour also contained abundant osteoclast-like giant cells, which raises the question of whether they were locally induced or whether they metastasised with the tumour.
Immunofluorescence microscopy for the rapid diagnosis of melioidosis.
Authors: Walsh A L , Smith M D , Wuthiekanun V , Suputtamongkol Y , Desakorn V , Chaowagul W , White N J
Published: June 2007

A direct immunofluorescent antibody test (DIF) was developed for the rapid diagnosis of melioidosis, a potentially fatal infection caused by Pseudomonas pseudomallei. In a clinical evaluation of 369 sputum, pus, or urine specimens from 272 patients with suspected melioidosis, the DIF had a sensitivity of 73% and a specificity of 99% compared with culture. Using this DIF, a confident diagnosis of melioidosis can now be made within two hours of admission to hospital, compared with the delay of two to four days required for culture results. Consequent early institution of specific antimicrobial therapy may help to save lives.
Hypercalcaemia in Hodgkin's disease related to prostaglandin synthesis.
Authors: Laforga J B , Vierna J , Aranda F I
Published: June 2007

A case of paraneoplastic hypercalcaemic syndrome is reported in a patient with Hodgkin's disease. This was detected eight months before widespread lymphadenopathy became apparent. Lymphocyte depleted Hodgkin's disease was diagnosed. PTH (parathyroid hormone) activity was suppressed and PTHRP (PTH related protein) was less than 5 pmol/l. 1,25(OH)2D3 was in the normal range. Plasma calcium values returned to normal after the administration of indomethacin. Thus the pathogenesis of the hypercalcaemia in this patient could be associated with the synthesis of prostaglandins.
Massive bone marrow necrosis and postnecrotic myelofibrosis in a patient with primary thrombocythaemia.
Authors: Majumdar G , Phillips J K , Pearson T C
Published: June 2007

A case of non-fatal massive bone marrow necrosis (MBMN) is reported in a patient with primary thrombocythaemia five years after diagnosis. No precipitating cause was identified. The patient developed postnecrotic myelofibrosis six months later. As far as is known, this is the first report of MBMN in a case of primary thrombocythaemia.
Histological features of CIN3 and their value in predicting invasive microinvasive squamous carcinoma.
Authors: al-Nafussi A I , Hughes D E
Published: June 2007

AIMS--To determine the histological features in CIN3 associated with or predictive of subsequent microinvasion. METHODS--The histological appearances of CIN3 accompanying 120 cases of microinvasive carcinoma of the uterine cervix were retrospectively studied. Major features were defined as those present in greater than 80% of cases of microinvasive carcinoma (MICA) and less than 10% of control cases of CIN3. One hundred cases of CIN3, 36 showing all, and 64 lacking all, of the major features associated with microinvasion, as defined in the retrospective study, were prospectively studied. Deeper levels were cut to exclude the presence of microinvasion in the original biopsy specimen and negative cases were followed up for a period of up to 18 months in order to assess rates of recurrence or progression. RESULTS--The major features identified in CIN3 associated with microinvasive carcinoma were extensive involvement of surface epithelium and deep endocervical crypts by expansile CIN3, luminal necrosis, and intraepithelial squamous maturation. Other features more commonly present in MICA associated CIN3 than in controls included frequent mitosis and apoptosis, pericryptal concentric fibroplasia, pericryptal inflammatory infiltrate, pronounced cellular pleomorphism, nuclear changes (distinct nucleoli and chromatin clearing), and the emergence of streams of darkly stained spindle cells oriented at right angles to the basement membrane. In the prospective study 83% of cases illustrating the major MICA associated features revealed evidence of MICA or frank invasion either on serial sections of the original biopsy or on subsequent biopsy. None of the 64 cases of CIN3 that lacked these features showed evidence of invasion on serial sections or on further follow up over 18 months. CONCLUSIONS--The data strongly suggest that cases of CIN3 which have a higher probability of association with or rapid progression to invasive disease can be identified. When these features are present in a biopsy specimen of CIN3, serial sections should be performed to exclude the presence of microinvasion. Closer clinical follow up of these patients may be needed.
C-erbB-2 immunostaining: problems with interpretation.
Authors: Kay E W , Walsh C J , Cassidy M , Curran B , Leader M
Published: June 2007

AIMS--To assess the consistency and reproducibility of assessment of c-erbB-2 immunostaining, and to examine some of the problems relating to inter- and intraobserver variability in the documentation of positive staining; to profile the spectrum of cytoplasmic and membranous staining in a wide range of tumour types. METHODS--A total of 283 neoplasms were examined for immunohistochemical expression of the c-erbB-2 oncoprotein. Three independent observers were required to assess intensity both of membrane and cytoplasmic staining on a three point and then a four point scale. Extent of positive staining was also assessed on a two point scale. A minimum of two weeks elapsed between assessments using the differing scales. RESULTS--Positive membrane staining was documented by one or more observers in 16.6% of tumours examined. This positivity was largely restricted to bladder, renal, and breast tumours. The overall level of disagreement as to the presence or absence of membranous staining was 11.3%. Cytoplasmic staining was identified in 55.5% of tumours studied. The level of disagreement as to the presence or absence of cytoplasmic staining was 26.5%. CONCLUSIONS--Intraobserver variability was minimal, indicating that each pathologist was adhering to internal reproducible standards. Interobserver variability was greater, indicating that the interpretation of c-erbB-2 immunostaining may require set guidelines. It is suggested that assessment should be referenced to a standard positive control, that a three tier system for grading of intensity and a two tier system for grading of extent should be adopted, and that the evaluation should be agreed by at least two pathologists. The presence of cytoplasmic staining should continue to be routinely recorded until its biological role and clinical implications are fully understood.
In situ end-labelling, light microscopic assessment and ultrastructure of apoptosis in lung carcinoma.
Authors: Gaffney E F , O'Neill A J , Staunton M J
Published: June 2007

AIMS--To compare in situ end-labelling (ISEL) of apoptosis in lung carcinoma with quantitative and semiquantitative light microscopic assessment and ultrastructural observations. METHODS--ISEL of apoptosis was evaluated in 42 lung carcinomas (24 squamous cell carcinomas, 12 adenocarcinomas and six small cell carcinomas). Results were correlated semiquantitatively with the extent of apoptosis in haematoxylin and eosin stained sections, with apoptotic indices and with ultrastructural observations (nine cases). RESULTS--In each tumour type the extent of apoptosis identified by ISEL correlated with that observed on light and electron microscopy. Tumour cells undergoing apoptosis showed either uniform nuclear staining with a surrounding "halo" or peripheral nuclear membrane staining. The latter pattern was more prominent in small cell carcinoma and correlated ultrastructurally with early apoptosis. A variable proportion of apoptotic cells and apoptotic bodies were unlabelled. Necrotic tumour cells were weakly stained but were distinguishable from apoptotic cells. CONCLUSIONS--ISEL, if used in conjunction with standard methods for investigating apoptosis, is a useful adjunct to the investigation of apoptosis in human tumour tissue.
Lymphoepithelial carcinoma of the salivary gland: in situ detection of Epstein-Barr virus.
Authors: Leung S Y , Chung L P , Yuen S T , Ho C M , Wong M P , Chan S Y
Published: June 2007

AIM--To examine the role of Epstein-Barr virus (EBV) in lymphoepithelial carcinoma of the salivary gland in Hong Kong Chinese. METHODS--Ten cases of lymphoepithelial carcinoma of the salivary gland (eight parotid and two submandibular) were examined. In situ hybridisation was used to localise EBER RNA, immunohistochemical methods to detect expression of latent membrane protein 1 (LMP-1) in EBV positive tumours, and Southern blot analysis to examine the clonality of EBV in the two cases where frozen tissue was available. RESULTS--None of the cases had a history of Sjögren's syndrome or histological evidence of a benign lymphoepithelial lesion. The IgA antibody titre against EBV viral capsid antigen was elevated in four cases. All cases were EBV positive by in situ hybridisation, with a strong uniform positive signal in the epithelial cells, and all cases expressed LMP-1. Southern blot analysis revealed that the clonal episomal form of the virus was present. Two of the three female patients in this series also developed carcinoma of cervix. One of these carcinomas had histological features of a lymphoepithelioma-like carcinoma but was EBV negative. CONCLUSIONS--A consistent association between EBV and lymphoepithelial carcinoma of the salivary gland was found. The presence of the virus in a clonal episomal form, and the expression of LMP-1 viral oncoprotein is further evidence of the role of EBV in the oncogenesis of this tumour.
Screening criteria for beta thalassaemia trait in pregnant women.
Authors: Rogers M , Phelan L , Bain B
Published: June 2007

AIMS--To establish suitable screening criteria for beta thalassaemia trait during pregnancy using an automated blood counter incorporating light scattering technology. METHODS--Pregnant women (n = 857) at a London antenatal clinic were investigated for beta thalassaemia trait if the Technicon H.2 full blood count showed either a mean corpuscular volume (MCV) < 85 fl or a mean corpuscular haemoglobin (MCH) < 27 pg. Results were then analysed to establish which of these variables was more suitable for screening and to determine suitable cut off points for calculating the haemoglobin A2 percentage. RESULTS--The MCH was superior to the MCV for thalassaemia screening as it was a more stable measurement and fewer unnecessary tests were performed. A MCH less than 27 pg is a suitable cut off point for screening. This screening criterion was equally applicable to a Coulter impedance counter. CONCLUSIONS--Pregnant women presenting at an antenatal clinic with a MCH < 27 pg should be investigated further to confirm or exclude a diagnosis of beta thalassaemia trait.
Ceroid granulomas in the female genital system.
Authors: Ooi K , Riley C , Billson V , Ostör A G
Published: June 2007

Three cases of ceroid granulomas of the female genital system are presented, involving the cervix in two and lesions in the ovaries and bowel serosa in the other. Ceroid granulomas are unusual and interesting lesions formed when suitable substrates accumulate within macrophages to such an extent that a relative lack of biological antioxidants results and auto-oxidation and conversion to ceroid is favoured. This may occur in the setting of haemorrhage and necrosis, whether from tumour necrosis or associated with endometriosis. Other sources of lipids and lipoproteins include bile, meconium and vernix caseosa.
Immunohistochemical detection of p53 and Bcl-2 proteins in Hashimoto's thyroiditis and primary thyroid lymphomas.
Authors: Chetty R , O'Leary J J , Biddolph S C , Gatter K C
Published: June 2007

AIMS--To investigate whether immunohistochemical staining using p53 and/or bcl-2 distinguishes between florid Hashimoto's thyroiditis and low grade mucosa associated lymphoid tissue (MALT) lymphoma of the thyroid. METHODS--Ten cases of Hashimoto's thyroiditis and eight of primary thyroid lymphoma were stained with monoclonal antibodies directed against p53 and bcl-2. RESULTS--In Hashimoto's thyroiditis most small lymphoid cells in mantle zones, within the thyroid parenchyma and in lymphoepithelial lesions expressed bcl-2 protein. Very occasional centroblasts in reactive germinal centres were positive for p53, but all other lymphoid cells from cases of Hashimoto's disease were negative for p53. In diffuse, low grade lymphomas bcl-2 protein was uniformly expressed by most tumour cells. However, low grade lymphomas with a follicular pattern did not express bcl-2. The diffuse, low grade lymphomas were negative for p53, while occasional larger cells in the follicular subtype were positive. Both high grade lymphomas were bcl-2 negative but strongly p53 positive. CONCLUSIONS--This study indicates that there is an inverse correlation between p53 and bcl-2 immunostaining in thyroid lymphomas (low grade lymphomas: bcl-2 positive, p53 negative; high grade lymphomas: bcl-2 negative, p53 positive). Furthermore, immunohistochemical staining for bcl-2 and p53 proteins does not distinguish florid Hashimoto's thyroiditis from diffuse, low grade thyroid lymphoma.
B cell signet-ring cell lymphoma of bone marrow.
Authors: McCluggage W G , Bharucha H , el-Agnaf M , Toner P G
Published: June 2007

A case of signet-ring cell lymphoma affecting the bone marrow and diagnosed by bone marrow trephine biopsy is reported. Normal marrow was replaced totally by cells with large central vacuoles, many of which displaced the nucleus to the periphery of the cell, imparting a signet-ring appearance. Initially, the favoured morphological diagnosis was metastatic signet-ring adenocarcinoma, but on immunocytochemistry the tumour cells were strongly positive for CD45 (leucocyte common antigen) and the B cell marker CD20 (L26). Electron microscopy revealed electron-lucent vacuoles with no discernable internal structure. The tumour was classified as a high grade centroblastic lymphoma using the upgraded Kiel classification. Despite chemotherapeutic treatment, the patient died during an episode of septicaemic shock within two months of presentation.
Desmin expression in rhabdomyosarcoma: influence of the desmin clone and immunohistochemical method.
Authors: Pollock L , Rampling D , Greenwald S E , Malone M
Published: June 2007

AIM--To determine which, if any, of five commercially available desmin clones is most reliable at labelling desmin filaments and whether the enhanced polymer one step (EPOS) method of labelling is of any advantage in the routine diagnostic laboratory. METHODS--Thirty four rhabdomyosarcomas from the files at The Hospital for Sick Children, Great Ormond Street, London, were studied. Four different desmin clones, DE-R-11, D33, DE-U-10, and PDE, were applied to each using the conventional extravidin biotin peroxidase method. The D33 clone was also applied using the EPOS method. RESULTS--The EPOS method incorporating D33 persistently scored more cells as desmin positive and was positive in four cases which were negative on staining with the other clones. CONCLUSIONS--The D33 desmin clone used with the EPOS method is more reliable for identifying desmin filaments in tumours than other desmin antibodies tested. Different desmin clones using a routine technique label different rhabdomyosarcoma cells and therefore it is justifiable to use more than one clone.
Histological findings in gastric mucosa in patients treated with non-steroidal anti-inflammatory drugs.
Authors: Caselli M , LaCorte R , DeCarlo L , Aleotti A , Trevisani L , Ruina M , Trotta F , Alvisi V
Published: June 2007

AIMS--To identify distinguishing and general histological features related to the use of non-steroidal anti-inflammatory drugs (NSAID). METHODS--Slides from gastric antral biopsies of 50 patients with osteoarthritis taking NSAID were compared with slides from antral biopsies of 50 control cases matched for age, sex, and race. Semithin sections stained with toluidine blue were used. RESULTS--Chronic gastritis was seen in 76% of the patients taking NSAID and in 58% of the control cases; active inflammation was detected in 10% of the NSAID treated patients and in 24% of the control cases, and it appeared closely related with Helicobacter pylori infection. Some histological features common to all slides of patients taking NSAID were recognised. These consisted of focal erosions of the gastric epithelium and macroerosions, and they seemed to represent successive steps of a process of "desquamation". CONCLUSIONS--Some distinguishing morphological aspects appeared prominent; it is suggested that these may be related to the pathogenesis of NSAID linked peptic ulceration. On the other hand, epithelial damage due to NSAID appears very different from that due to Helicobacter pylori, another important factor involved in the aetiopathogenesis of peptic disease.
Splenic lymphoma with circulating villous lymphocytes.
Authors: Imbing F , Kumar D , Kumar S , Yuoh G , Gardner F
Published: June 2007

This report describes the occurrence of splenic lymphoma with villous lymphocytes (SLVL) in a 56 year old white female with a family history of chronic lymphocytic leukaemia. Other unusual features included a marked lymphocytosis with counts up to 224 x 10(9)/l and marked clumping of lymphocytes in EDTA anticoagulated blood. The neoplastic cells were CD19+, CD20+, CD22+, CD22+, IgM+, lambda+, kappa-, CD5-, and CD10-. The spleen had nodular infiltrates of B lymphocytes in the region of the white pulp with minimal red pulp involvement. Electron microscopy of peripheral blood lymphocytes revealed cells with polar cytoplasmic processes. This report underlines the need for detailed analysis, including morphology and immunophenotyping, for each patient with a small B cell lymphoproliferative disorder.
Production and characterisation of an antimelanoma monoclonal antibody KBA.62 using a new melanoma cell line reactive on paraffin wax embedded sections.
Authors: Cohen-Knafo E , al Saati T , Aziza J , Ralfkiaer E , Selves J , Gorguet B , Delsol G
Published: June 2007

AIMS--To generate new monoclonal antibodies directed against melanoma associated antigens using a new melanoma cell line, KAL. METHODS--The melanoma cell line was established in culture from a lymph node metastasis of malignant melanoma. Normal Balb/c mice were immunised with KAL cells. Splenocytes were used for fusion experiments using standard techniques. Hybridoma supernatants were tested for antibody binding activity using an indirect immunoperoxidase method on frozen sections from KAL tumour cells xenografted onto nude mice and human tonsils. KBA.62 was selected because of its reactivity with melanocytic proliferations on both frozen and paraffin wax sections. RESULTS--On immunoblotting, KBA.62 reacted with three bands of 140, 135 and 128 kD and two weak bands of 88 and 73 kD. In normal human tissues basal melanocytes in the epidermis did not react with this antibody and only occasional labelling of endothelial cells was noted. Of the human tumours, KBA.62 reacted strongly and uniformly with the majority of benign (21/21) and malignant (75/86) melanocytic proliferations. Staining was localised predominantly to the cell membrane with little or no cytoplasmic reactivity. Negative staining was observed in the majority of human non-melanocytic neoplasms, the exceptions being some carcinomas (11/89), particularly the well differentiated squamous cell type. This, however, was not thought to present a diagnostic problem. CONCLUSIONS--KBA.62 appears to be potentially useful in ascertaining the immunomorphological diagnosis of malignant melanoma in routinely processed paraffin wax sections.
Management of blood donors whose donations are repeatedly falsely positive by the HIV antibody screening test.
Authors: Atrah H I , Parry J V , Gough D , Tosswill J , Ala F A
Published: June 2007

Since 1985, over 1,800,000 donations have been screened by the West Midlands Regional Blood Transfusion Service for antibody to HIV. Twelve regular donors gave three or more donations that were alternatingly positive and negative in the screening test, but not confirmed to be HIV positive by supplementary testing. Extensive investigation of six of these donors, including the polymerase chain reaction (PCR), failed to confirm HIV infection. The donors were reassured but, nevertheless, retired to comply with the guidelines of the National Blood Transfusion Service. These findings indicate that, for UK donors, ambiguous serological findings are unlikely to reflect HIV infection. On the rare occasions where serological results are particularly ambiguous, PCR testing of donors' blood may be helpful.
Combined goblet cell carcinoid and mucinous cystadenoma of the appendix.
Authors: al-Talib R K , Mason C H , Theaker J M
Published: June 2007

Two cases of combined goblet cell carcinoid and mucinous cystadenoma occurring in the appendix are reported. The histogenesis of the goblet cell carcinoid remains one of its most controversial aspects and the occurrence of both of these relatively uncommon tumours in the same organ may lend support to the unitary stem cell hypothesis on the origin of this tumour. Alternatively, this occurrence may represent an example of the adenoma/carcinoma sequence.
Detection and widespread distribution of Chlamydia pneumoniae in the vascular system and its possible implications.
Authors: Ong G , Thomas B J , Mansfield A O , Davidson B R , Taylor-Robinson D
Published: June 2007

AIMS: To attempt to detect Chlamydia pneumoniae DNA in atheromatous vascular tissue. METHODS: A modification of an existing polymerase chain reaction (PCR) assay and immunofluorescence staining with a monoclonal antibody directed against C pneumoniae were used to detect C pneumoniae. Specimens from 32 patients undergoing abdominal aortic aneurysm repair were examined. Vascular tissue, ostensibly normal, from six liver transplant donors was also examined for comparison. Altogether, 43 vessels from these 38 subjects (age range 36-85 years) were examined. RESULTS: C pneumoniae was detected in 11 (44%) of 25 aortas, five (55%) of nine iliac arteries, two (40%) of five femoral arteries, and one of two iliac veins. Immunofluorescence staining supported positive PCR results in three of 12 cases in which it was used. Overall, C pneumoniae was detected in the arteries of 14 (44%) of the patients undergoing vascular surgery and three (50%) of the donors. CONCLUSIONS: This study is the first in the UK in which C pneumoniae organisms have been found in atherosclerotic vessels and the tendency for the organisms to be present most often in such vessels exhibiting chronic inflammatory changes suggests that a search for them in various forms of arteritis may also be rewarding.
Comparison of the ligase chain reaction with cell culture for the diagnosis of Chlamydia trachomatis infection in women.
Authors: Ridgway G L , Mumtaz G , Robinson A J , Franchini M , Carder C , Burczak J , Lee H
Published: June 2007

OBJECTIVE: To determine the sensitivity and specificity of ligase chain reaction (LCR) analysis of cervical and urine specimens from women compared with cell culture of cervical and urethral specimens for the diagnosis of genitourinary chlamydial infection. METHODS: Women (n = 624) attending the Genitourinary Medicine Clinic at University College London Hospitals, were enrolled. Patients who had received antibiotics within the previous two weeks were excluded. Specimens were obtained from the urethra and cervix for chlamydial culture, and from the cervix for LCR. A specimen of first void urine was also obtained for LCR. Discrepancies were resolved by direct immunofluorescence or a major outer membrane protein targeted LCR, or both. RESULTS: The prevalence of Chlamydia trachomatis in 600 patients, using an expanded standard of a positive cell culture or two confirmed positive non-culture tests, was 13.2% (79/600). Cervical culture detected 68.4% and urethral culture 62% of all positive results compared with 81% detected by cervical LCR and 69% by urine LCR. Cervical and urethral culture combined detected 87.3% whereas cervical and urine LCR combined detected 91.1% of positive cases. Specificity of LCR was 100% in the cervix and 99.8% in urine. CONCLUSION: This study demonstrates that LCR analysis of cervical and urine specimens is a reliable method for the diagnosis of chlamydial genital infection in women. However, the study also demonstrates that no single test will detect all chlamydial infections. Conventional non-culture tests and cell culture may grossly underestimate the prevalence of chlamydial infection. LCR analysis of a cervical specimen was superior to conventional cell culture without blind passage as a single test for diagnosing chlamydial infection in women, followed by LCR of a urine specimen.
Measurement of extent of spread of oesophageal squamous carcinoma by serial sectioning.
Authors: Lam K Y , Ma L T , Wong J
Published: June 2007

OBJECTIVES: (1) To examine the prevalence and extent of intramural metastasis in squamous cell carcinomas of the oesophagus so as to delineate the resection margins for these tumours; (2) to devise an appropriate method for measurement of these lesions which takes into account of the contraction of the specimens after resection. METHODS: Oesophagectomy specimens were prospectively collected from 96 patients (87 males, nine females) with primary oesophageal squamous cell carcinoma over a two year period. The sizes of the tumours were measured in situ, after resection and after application of muscle relaxant (to regain their in situ length). The specimens were then serially sectioned for histological examination. RESULTS: The sizes of the tumours measured after application of muscle relaxant roughly corresponded to those measured in situ. Intramural metastasis was observed in 26% of the cases. Sixty four per cent (16 cases) of these were on the oral side, 72% (18 cases) on the gastric side, and 25% (nine cases) on both sides of the tumours. The most distant extent of intramural metastasis from the primary tumour was from 0.5 cm to 7.7 cm (mean = 3.4 cm) on the oral side, and 0.5 to 9.5 cm (mean 4 cm) on the gastric aspect of the tumour. Intramural metastasis was seen only in patients in whom the primary cancer had deep muscle infiltration. Multiple neoplastic lesions could be detected in 33% of the patients. Both intramural metastasis and multiple neoplastic lesions were associated with extensive lymph node infiltration. However, they had different histological features and extent of infiltration. CONCLUSIONS: Intramural metastasis was frequently observed in oesophageal squamous cell carcinoma. This implies that excision with wide margins should be considered for local control of the disease.