bluuebunny/crossref_metadata_2025 huggingface dataset:
Phenotypic methods for speciating clinical Aeromonas isolates.
Authors: Wilcox M H , Cook A M , Thickett K J , Eley A , Spencer R C
Published: June 2007

AIMS: To establish the suitability of currently available phenotypic methods for speciation of clinical Aeromonas isolates in diagnostic microbiology laboratories. METHODS: Using 62 Aeromonas spp, three schemes based on biochemical reactions were compared: a series of conventional tests; a system based on the suicide phenomenon, comprising two tubes in total; and a commercially available test, API 20 NE, augmented with a plate assay for beta haemolysin production. The whole cell and outer membrane protein (OMP) profiles of strains were examined by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS PAGE), according to the results of the above schemes, to determine the intra-species homogeneity. RESULTS: Ninety per cent of strains were identified satisfactorily according to conventional criteria. For these strains, agreement was obtained using the suicide phenomenon and API schemes in 93% and 88% of cases, respectively. The three schemes concurred for 82% of strains. Whole cell protein profiles were unsuitable for comparing strains within a species. However, OMP patterns were similar for 89% of A caviae and 63% of A hydrophila. CONCLUSION: Phenospeciation of clinical Aeromonas isolates by the scheme based on the suicide phenomenon is simple to perform and accurate, and suitable for use in the diagnostic laboratory. OMP profiles are potentially useful for confirming the identity of A caviae and most A hydrophila, but not A sobria.
Audit of bone marrow trephines.
Authors: Bishop P W , McNally K , Harris M
Published: June 2007

AIMS: To establish criteria of adequacy for bone marrow trephine biopsy specimens and to audit the quality of trephines performed at the Christie Hospital, Manchester. METHODS: Trephines (n = 767) performed over 12 months were reviewed. Their lengths, and the lengths of their constituent parts (soft tissue, cortex, crushed marrow and interpretable marrow) were measured. The mean performance of each operator was calculated. Criteria of adequacy were established by a review of the published findings and an analysis of the relation between trephine length and the rate of infiltration by tumour. RESULTS: Before processing, the average trephine was 1.59 cm long. Trephines shrunk by 25% during processing. In histological sections the average length was 1.15 cm, consisting of 0.09 cm of soft tissue, 0.04 cm of cortex, 0.26 cm of disrupted marrow and 0.74 cm of interpretable marrow. A large number of operators were taking trephine biopsy specimens and their performance varied considerably. Review of the published findings suggested that the minimum adequate length is in the range 1.5 cm to 2.0 cm. The analysis of the relation between length of trephine and the rate of positivity for neoplasia yielded a minimum adequate length of 1.2 cm in section (1.6 cm before processing). Fifty eight per cent of the trephines were inadequate by this criterion. There was a tendency for the Jamshidi needle to produce a longer trephine than the Islam needle. CONCLUSION: According to objective criteria, at the Christie Hospital, many operators are producing a high proportion of inadequate bone marrow trephines.
Measurement of placental alkaline phosphatase activity in benign and malignant pleural effusions.
Authors: Fergusson R J , Fisken J , McIntyre M A , Roulston J E , Leonard R C
Published: June 2007

The usefulness of placental alkaline phosphatase (PLAP) as a diagnostic marker of malignancy was assessed in pleural fluid from 60 patients with effusions. Pleural fluid PLAP activities were measured by an enzyme linked immunoassay (ELISA) using the two monoclonal antibodies H17E2 and H317. Similar values were found in groups of patients with primary bronchial tumours (n = 12), secondary malignancies (n = 23), and "benign" conditions (n = 25). The highest values were found in a small subgroup of patients with metastatic ovarian carcinoma. However, the production of this enzyme by normal lung makes the measurement of PLAP in pleural fluid unhelpful as a diagnostic aid to distinguish "benign" from malignant effusions.
Eosinophilic bodies in pyloric and Brunner's gland cells.
Authors: Rubio C A , Hirota T , Itabashi M , Jacobsson B , Lignelid H
Published: June 2007

A previously unreported cell phenotype occurred in the pyloric and Brunner glands in two gastrectomy specimens. The cells were characterised by homogeneous, eosinophilic material in the cytoplasm. The eosinophilic material had an abnormally strong reactivity for Cystatin C, a protein found recently in the normal secretion of pyloric and Brunner's gland cells. The reason for the apparent cytoplasmic accumulation of cystatin C in the two patients described remains unclear.
Reassessment of the rate of fixative diffusion.
Authors: Start R D , Layton C M , Cross S S , Smith J H
Published: June 2007

The diffusion of fixatives is slow. Early work using plasma gels and animal tissues showed the distance penetrated by a fixative to be a simple function of the fixation time but this relation has not been established in human tissues. The rates of diffusion into whole human spleens were measured for three primary fixatives over periods ranging from one to 25 days. A positive correlation was demonstrated between penetration distance (mm) and fixation time (hours). The diffusion rates were slower than those in previous studies. These results have possible implications for the handling of surgical specimens.
Quality control in immunocytochemistry: experiences with the oestrogen receptor assay.
Authors: Bosman F T , de Goeij A F , Rousch M
Published: June 2007

AIMS: To evaluate the feasibility of an interlaboratory quality control programme in immunohistochemistry. METHODS: Several pathology laboratories were asked to carry out immunohistochemical oestrogen receptor staining on a set of freeze dried cryostat sections of breast cancer tissue. The sections and protocols for staining and semi-quantitative scoring were mailed to the participating laboratories in two trials. The oestrogen receptor content of the breast cancer samples was determined by radioligand binding assay on the tumour cytosol. RESULTS: In the first trial 11 laboratories (response rate 60%) participated. Eight (73%) of the participants scored within a 95% confidence interval and all but one correctly classified the tumour as receptor positive. In the second trial all 20 participating laboratories (response rate 55%) correctly scored one tumour sample as negative and 18 of them (90% of respondents) correctly classified the two other tumour samples as receptor positive. In a quantitative evaluation a histochemical score within 95% confidence interval limits was provided by eight (40%) and 12 (60%) of the participants. CONCLUSIONS: Semiquantitative scoring of immunocytochemical staining is valuable for performing correlative inter-laboratory studies, although this scoring protocol may not be required for diagnosis or prognosis. Significant inter-laboratory variability exists, leading to qualitatively correct receptor classification in 100% of receptor negative and 80% of receptor positive cases, and quantitative agreement in only about half of the cases. The perceived variability is not caused by systematic differences in the choice of the immunocytochemical technique, or the mailing of freeze dried sections. Quality control programmes should be included in the standard procedures of each diagnostic immunohistochemistry laboratory.
Use of monoclonal antibody E48 in diagnosing transitional cell carcinoma of urinary bladder.
Authors: Torenbeek R , Blomjous C E , Quak J J , Ybema S , Meijer C J
Published: June 2007

AIMS: To investigate the localisation of the E48 epitope and to determine the use of monoclonal antibody E48 for the identification of transitional cell carcinomas (TCC); and to determine if antigenic expression was affected by different standard fixation methods. METHODS: Biopsy specimens were labelled with E48 for immunoelectron microscopy. One hundred and nineteen tissue samples from 47 bladder carcinomas were tested for reactivity with E48, using fresh frozen, sublimate formalin, and formalin fixed tissue. Thirteen undifferentiated bladder tumours and 10 undifferentiated prostatic carcinomas were incubated with E48 and prostate specific antigen. RESULTS: Reactivity to E48 was found in all grade 1 and 2 carcinomas and most (83%) grade 3 tumours. At the ultrastructural level, expression was mainly associated with desmosomes and the cytoplasmic membrane. The reactivity of E48 was generally strong in fresh frozen tissue samples and remained preserved in fixed tissue samples. Ten of the 13 bladder carcinomas expressed E48; all prostatic tumours were totally negative. CONCLUSIONS: E48 is a sensitive marker for transitional cell carcinoma and suitable for differentiation between urothelial and prostatic undifferentiated carcinoma. It can be used in routinely processed, formalin fixed, biopsy specimens.
Differential effect of chronic hepatitis D virus infection on intrahepatic expression of hepatitis B viral antigen.
Authors: Lau J Y , Portmann B C , Alexander G J , Williams R
Published: June 2007

AIMS: To determine how chronic hepatitis D virus (HDV) infection affects intrahepatic hepatitis B virus (HBV) antigen expression. METHODS: Ninety eight liver biopsy specimens from 68 patients seropositive for total antibody to HDV were studied by immunohistochemistry, and the amount of HBV antigens was also quantified by radioimmunoassay in 12 patients and compared with 30 patients with chronic HBV infection. RESULTS: Forty nine of the 68 patients were positive for intrahepatic HDV antigen and only five were positive for HBV core antigen (HBcAg). HBV surface antigen (HBsAg) was present in 55 (80.9%) patients and was always cytoplasmic in distribution. Hepatic pre-S1 and pre-S2 expressions paralleled that of HBsAg, and were detected in 53 (77.9%) and 54 (79.4%) patients, respectively. There was no relation between the intrahepatic expression of HDV antigen and HBsAg/pre-S1/pre-S2. Follow up biopsy specimens in 25 patients showed either static or deteriorating histology while intrahepatic HDV antigen remained the same or fell. The patients with intrahepatic expression of HBcAg had either absent or noticeably decreased expression of HBcAg in their follow up biopsy specimens (median two years). In contrast, HBsAg/pre-S1/pre-S2 were the same or increased (p less than 0.001). Quantification of intrahepatic HBsAg in patients with chronic HDV infection (0.61 pg/hepatocyte, range: 0.05-1.08, n = 12) showed no difference with patients with chronic HBV infection alone (0.64 pg/hepatocyte, range: 0.02-1.02, n = 30, p = NS). CONCLUSION: These data indicate that chronic HDV infection suppresses intrahepatic expression of HBcAg but not HbsAg and pre-S antigens, suggesting a differential effect of chronic HDV infection on HBV gene expression.
Oncogene expression in primary myelodysplasia: correlation with haematological, karyotypic, and clinical progression.
Authors: Hutchinson R M , Pringle J H , Knight S C , Lauder I , Potter A , Jagger C
Published: June 2007

AIMS: To see if the relative expressions of proto-oncogenes that are increased in acute myeloid leukaemia are raised in patients with myelodysplastic syndromes (MDS), and to see if they increase with progression to leukaemia. To note if there is a correlation between morphology, karyotype, and these proto-oncogene expressions and if any one proto-oncogene can predict prognosis. METHOD: Bone marrow from 130 patients was analysed at six monthly intervals over two years for relative mRNA expression of seven oncogenes, karyotype, and morphology. The technique used slot blot hybridisation and densitometric analysis. The results were compared with 14 surgical controls and 30 people with vitamin deficiency anaemia. RESULTS: Six of seven oncogenes showed increased expression which progressed with time, but did not correlate with morphological or karyotypic changes. Expression of four of the seven oncogenes was increased in megaloblastic and iron deficiency anaemia. C-mos showed differences among the five morphological subgroups; it correlated with abnormal location (p = 0.025) and seemed to influence prognosis. CONCLUSION: Increased proto-oncogenes reflect the overall marrow perturbation in MDS. C-mos may reflect persistence of monocyte pathway which confirms marrow stability.
Acquired factor XI inhibitor in chronic lymphocytic leukaemia.
Authors: Goodrick M J , Prentice A G , Copplestone J A , Pamphilon D H , Boon R J
Published: June 2007

A 71 year old man with chronic lymphocytic leukaemia (CLL) experienced excessive bleeding following transurethral resection of the prostate. Investigations showed a prolonged kaolin cephalin clotting time (KCCT) with low concentrations of factor XI. The prolonged KCCT was largely corrected by mixing with normal plasma but this correction was lost on incubation, confirming the presence of an inhibitor. He was treated with pulsed methylprednisolone and chlorambucil which resulted in the resolution of the bleeding problem and the loss of detectable circulating inhibitor.
Solitary ganglioneuroma of the rectum: report of two cases.
Authors: Beer T W
Published: June 2007

Two cases of solitary rectal ganglioneuromas are reported, one in a patient with several previously resected colorectal adenomas, the other in a patient with no known predisposing pathology. No prior reports of cases of solitary rectal ganglioneuroma have been published as far as is known, and the origin of similar lesions which have been reported at other sites in the gastrointestinal tract is a subject for speculation.
Dieulafoy's vascular malformation as a cause of large intestinal bleeding.
Authors: Farrell D J , Bennett M K
Published: June 2007

Four cases of Dieulafoy's vascular malformation of the caecum are reported. Three were associated with massive large bowel haemorrhage, one of which was fatal. The remaining case had a calibre persistent submucosal artery within the caecum that was found incidentally in a resection specimen. This vascular malformation should be considered when dealing with specimens resected for massive lower gastrointestinal bleeding.
Urinary excretion of beta 2-glycoprotein-1 (apolipoprotein H) and other markers of tubular malfunction in "non-tubular" renal disease.
Authors: Flynn F V , Lapsley M , Sansom P A , Cohen S L
Published: June 2007

AIM: To determine whether urinary beta 2-glycoprotein-1 assays can provide improved discrimination between chronic renal diseases which are primarily of tubular or glomerular origin. METHODS: Urinary beta 2-glycoprotein-1, retinol-binding protein, alpha 1-microglobulin, beta 2-microglobulin, N-acetyl-beta-D-glucosa-minidase and albumin were measured in 51 patients with primary glomerular disease, 23 with obstructive nephropathy, and 15 with polycystic kidney disease, and expressed per mmol of creatinine. Plasma beta 2-glycoprotein-1 was assayed in 52 patients and plasma creatinine in all 89. The findings were compared between the diagnostic groups and with previously published data relating to primary tubular disorders. RESULTS: All 31 patients with plasma creatinine greater than 200 mumol/l excreted increased amounts of beta 2-glycoprotein-1, retinol-binding protein, and alpha 1-microglobulin, and 29 had increased N-acetyl-beta-D-glucosaminidase; the quantities were generally similar to those found in comparable patients with primary tubular pathology. Among 58 with plasma creatinine concentrations under 200 mumol/l, increases in beta 2-glycoprotein-1, retinol-binding protein, and alpha 1-microglobulin excretion were less common and much smaller, especially in those with obstructive nephropathy and polycystic disease. The ratios of the excretion of albumin to the other proteins provided the clearest discrimination between the patients with glomerular or tubular malfunction, but an area of overlap was present which embraced those with obstructive nephropathy and polycystic disease. CONCLUSIONS: Increased excretion of beta 2-glycoprotein-1 due to a raised plasma concentration or diminution of tubular reabsorption, or both, is common in all the forms of renal disease investigated, and both plasma creatinine and urinary albumin must be taken into account when interpreting results. Ratios of urinary albumin: beta 2-glycoprotein-1 greater than 1000 are highly suggestive of primary glomerular disease and those less than 40 of primary tubular disease. Used in this way, beta 2-glycoprotein-1 assays provide superior discrimination between glomerular and tubular malfunction when compared with retinol binding protein but the best discrimination is provided by albumin: alpha 1-microglobulin ratios.
Epithelioid haemangioendothelioma of soft tissue after pellet injury
Authors: Sarma N H H Nadha
Published: June 2007

An epithelioid haemangioendothelioma developed at the site of injury sustained from an air gun pellet 20 years previously in an otherwise healthy 30 year old man, suggesting a possible traumatic aetiology in this case.
A pragmatist's approach to pathology costing: the Welsh Datatree project.
Authors: Gozzard D I , Macaulay M E , Nuttall D S , Jones E R
Published: June 2007

The Datatree costing project in Wales has provided the Welsh pathology laboratories with a standard costing package that allows pathologists to understand how their own laboratory's test costs are compiled. The software provides answers to the question "what if? ..." and shows instantly the effect of salary or consumable cost alterations. Resource management at a laboratory level is enhanced by a greater knowledge of costs, particularly in relation to volumes of work. Perhaps this is one of the stepping-stones across the river to the "open market." In the United Kingdom NHS any information of this kind must be regarded as invaluable.
Non-isotopic in situ hybridisation and immunophenotyping of infected cells in the investigation of human fetal parvovirus infection.
Authors: Morey A L , Porter H J , Keeling J W , Fleming K A
Published: June 2007

AIMS: To compare the use of biotinylated and digoxigenin labelled probes for diagnosis of human fetal parvovirus B19 infection in formalin fixed, paraffin wax embedded tissues; and to assess the cellular distribution of the virus in positive cases. METHODS: Sections of lung tissue from 23 cases of anatomically normal non-immune fetal hydrops presenting between 1984 and 1989, and from 13 control cases of hydrops due to chromosomal abnormality were probed for B19 DNA by in situ hybridisation using both biotinylated and digoxigenin labelled probes. The distribution of the virus was then investigated in all cases of fetal B19 infection confirmed in this laboratory to date (n = 11) by combining in situ hybridisation for viral DNA (using the digoxigenin system) with immunohistological labelling for a range of cellular antigens. RESULTS: Five unequivocal cases of B19 infection were identified among the 23 fetuses with unexplained hydrops using both probe labels. When combined with data from previous studies of the period 1974-1983, the results indicate that B19 infection was responsible for 27% of cases of anatomically normal non-immune hydrops and 8% of all cases, of non-immune hydrops presenting to this hospital over 15 years. False positive signal was seen in an additional three cases, using biotinylated probes. Digoxigenin labelled probes gave greater specificity and permitted detailed investigation of tissues high in endogenous biotin. Though most cells containing B19 DNA colabelled as erythroid precursors, viral DNA was frequently detected within mononuclear-phagocytic cells. In three cases viral signal was also found within occasional myocardial cells labelled by antibody to desmin. CONCLUSIONS: A relatively high proportion of cases of anatomically normal, non-immune hydrops are caused by B19 infection. Digoxigenin is a more reliable probe label than biotin for in situ hybridisation in archival fetal tissues. Double labelling for cellular antigens and viral nucleic acid is a powerful technique for investigating virus-host cell interactions, and provides evidence that cell types other than those of erythroid lineage may have a role in human fetal parvovirus infection.
Glutathione S-transferases in neonatal liver disease.
Authors: Mathew J , Cattan A R , Hall A G , Hines J E , Nelson R , Eastham E , Burt A D
Published: June 2007

AIMS: To investigate the distribution of alpha and pi class glutathione S-transferases (GST) in normal fetal, neonatal, and adult liver; and to examine changes in GST expression in neonatal liver disease. METHODS: alpha and pi class GST were immunolocalised in sections of formalin fixed liver tissue obtained from human fetuses (n = 21), neonates (n = 8), young children (n = 9) and adults (n = 10), and from neonates with extrahepatic biliary atresia (n = 15) and neonatal hepatitis (n = 12). Monospecific rabbit polyclonal antibodies were used with a peroxidase-antiperoxidase method. RESULTS: Expression of pi GST was localised predominantly within biliary epithelial cells of developing and mature bile ducts of all sizes from 16 weeks' gestation until term and in neonatal and adult liver. Coexpression of pi and alpha GST was seen in hepatocytes of developing fetal liver between 16 and 34 weeks' gestation. Although pi GST was seen in occasional hepatocytes up to six months of life, this isoenzyme was not expressed by hepatocytes in adult liver. By contrast, alpha GST continued to be expressed by hepatocytes in adult liver; this isoenzyme was also seen in some epithelial cells of large bile ducts in adult liver. No change was observed in the distribution of alpha GST in either neonatal hepatitis or extrahepatic biliary atresia. However, aberrant expression of pi GST was identified in hepatocytes of all but one case of extrahepatic biliary atresia but in only two cases of neonatal hepatitis. CONCLUSIONS: The phenotypic alterations noted in extrahepatic biliary atresia may result from the effect of cholate stasis. Evaluation of the pattern of pi and alpha GST distribution by immunohistochemical staining may provide valuable information in distinguishing between these two forms of neonatal liver disease.
Collagen remodelling in myocardia of patients with diabetes.
Authors: Shimizu M , Umeda K , Sugihara N , Yoshio H , Ino H , Takeda R , Okada Y , Nakanishi I
Published: June 2007

AIMS: To investigate collagen remodelling in the interstitium of the heart in patients with diabetes. METHODS: Immunohistochemical study of the biopsied myocardium using type specific anticollagen antibodies (I, III, IV, V, VI) was performed in 12 patients with non-insulin dependent diabetes mellitus and six non-diabetic patients. There was no history of hypertension or coronary artery stenosis in any of the patients. RESULTS: Noticeable accumulations of collagen types I, III, and VI in the myocardial interstitium were recognised in both groups, but little accumulation of types IV or V was found. Types I and III mainly stained in the perimysium and perivascular region, while type VI predominantly stained in the endomysium. There was no disease specific accumulation of collagen in diabetes mellitus. The percentage of total interstitial fibrosis in the myocardium was significantly higher in the diabetic group than in the control group (p < 0.05). Although the percentages of collagen types I and VI did not differ between the two groups, the percentage type of III was significantly higher in the diabetic group than in the controls (p < 0.01). CONCLUSIONS: Collagen remodelling mainly as a result of an increase in collagen type III in the perimysium and perivascular region, occurs in the hearts of patients with diabetes.
Intramural platelet deposition in cerebral vasculopathy of systemic lupus erythematosus.
Authors: Ellison D , Gatter K , Heryet A , Esiri M
Published: June 2007

AIMS: To test the hypothesis that fragments of platelet thrombi and vascular endothelium are incorporated into the walls of small cerebral vessels in systemic lupus erythematosus (SLE). METHODS: Six varied necropsy cases of central nervous system (CNS) SLE and 15 controls were studied. The controls were selected to represent a wide range of diseases in which the cerebral circulation is compromised. Tissue sections were stained by standard histochemical and immunocytochemical methods, the latter using antibodies to platelet membrane glycoprotein IIIa (CD61), and vascular endothelium (CD31). RESULTS: In four of six cases of CNS SLE characterised by small vessel hyalinization and thickening, fragments of platelet membrane were found in the walls of small cortical and meningeal vessels. Similar findings were not evident in two other SLE cases that were characterised by relatively short clinical histories and an acute vasculitis. One control case of severe polyarteritis nodosa showed platelet fragment deposition in arteries larger than the vessels so affected in SLE. CONCLUSIONS: Previous studies have suggested that neuropsychiatric symptoms in SLE may be related to repeated episodes of vasculitis in small cerebral vessels that are triggered by antiphospholipid antibodies. Concurrent thrombus formation might facilitate the incorporation of platelet fragments into small vessel walls. This process contributes to the thickening and irregularity of small vessels, a major feature of longstanding cases of CNS SLE.
Immunological and ultrastructural studies in acute biphenotypic leukaemia.
Authors: Shetty V , Chitale A , Matutes E , Buccheri V , Morilla R , Catovsky D
Published: June 2007

AIMS--To compare the sensitivity of the ultrastructural method to detect myeloperoxidase (MPO) with light microscopy and immunocytochemistry using an anti-MPO antibody; to examine the expression of lymphoid antigens in relation to MPO activity in blast cells from cases of biphenotypic leukaemia. METHODS--Blast cells from 14 cases of biphenotypic acute leukaemia were analysed. Immunological markers were performed by single or double immunofluorescence staining on a flow cytometer. The presence of MPO was determined by light microscopy, electron microscopy on fixed and unfixed cells, and by immunoalkaline phosphatase with an anti-MPO antibody. The immunogold method was applied at the ultrastructural level to assess the expression of lymphoid and myeloid antigens at the same time as the MPO activity. RESULTS--Six of the 14 cases were initially classified as acute lymphoblastic leukaemia (ALL) and eight as acute myeloid leukaemia (AML). MPO activity was shown at the ultrastructural level in 4-99% blasts from all cases. Six of the 14 were MPO negative by light microscopy and three of these were negative with the antibody anti-MPO. Coexpression of lymphoid antigens (CD19, CD10, or CD2) and MPO was shown by the immunogold method in four out of 11 cases; in seven cases the blasts coexpressed myeloid antigens (CD13, CD33) and MPO. CONCLUSIONS--Electron microscopy is more sensitive for showing MPO than light microscopy and immunocytochemistry; the immunogold method combined with MPO used at the ultrastructural level can help to define the cell lineage involved in biphenotypic leukaemia by highlighting the myeloid component defined by MPO.
Evaluation of sensitivity of 10 diagnostic assays for Chlamydia trachomatis by use of a simple laboratory procedure.
Authors: Thomas B J , MacLeod E J , Taylor-Robinson D
Published: June 2007

AIMS--To determine the sensitivity of commercially available diagnostic assays for Chlamydia trachomatis using a simple method. METHODS--Nine commercial assays and an "in-house" polymerase chain reaction (PCR) were evaluated using serial dilutions of a laboratory grown H serovar--four of them using a laboratory grown E serovar. Seven of the assays were further tested using dilutions of several cervical samples known to contain chlamydiae. RESULTS--The most sensitive assays were the MicroTrak direct fluorescent antibody (DFA) test (Syva) and the PCR which detected C trachomatis at a 10(-8) dilution of the H serovar, while the two least sensitive, Clearview (Unipath) and TestPack (Abbott), were positive only at 10(-4) and 10(-3) dilutions, respectively. A range of enzyme immunoassays (EIAs) and a nucleic acid hybridisation test were of intermediate sensitivity. The results with serovar E were consistent with these. When clinical samples were examined, the DFA test detected C trachomatis in dilutions at least 10-fold greater than any other assay. CONCLUSIONS--The range of sensitivity of diagnostic assays determined by the laboratory dilution procedure is very wide. Sensitivity assessed in this way, however, reflects the ability of the assays to detect C trachomatis in large scale clinical trials. The dilution procedure, which is simple to undertake, could therefore be applied by any laboratory before a new diagnostic method is considered for routine use.
Haemopoietic regrowth after chemotherapy for acute leukaemia: an immunohistochemical study of bone marrow trephine biopsy specimens.
Authors: Wilkins B S , Bostanci A G , Ryan M F , Jones D B
Published: June 2007

AIM--To analyse haemopoietic regrowth and residual disease in bone marrow trephine biopsy specimens after treatment for acute leukaemia, using immunohistochemical staining. METHODS--Biopsy specimens before and after treatment were studied from patients diagnosed as having acute myeloid or lymphoblastic leukaemia. Specimens after treatment encompassed periods from two to 56 weeks from the start of treatment. Routine haematoxylin and eosin and Giemsa stained sections were evaluated in association with immunostained preparations. A panel of antibodies was used, which reacts with epitopes showing restricted expression dependent on the lineage or maturation stage of cells. Results were evaluated in the light of clinical, peripheral blood, and marrow aspirate findings. RESULTS--The speed and sequence of regrowth of haemopoietic cells were more variable than expected. Immunostaining highlighted features of dysplasia after treatment and in some cases assisted detection of residual or relapsed leukaemia. Peripheral blood and aspirate cell counts reflected accurately the amount of regrowth, but not the dysplasia, seen in biopsy samples. Delayed regrowth was associated with complex individual factors. CONCLUSIONS--Morphological and immunohistochemical study of trephine biopsy specimens from patients treated for acute leukaemia provides information complementary to that obtained from peripheral blood and aspirated marrow. Variation in the timing and sequence of regrowth is highlighted. Immunostaining can aid in the detection of relapse or minimal residual leukaemia. The clinical relevance of dysplastic changes in biopsy specimens after treatment is uncertain, but such changes may persist for long periods.
Expression of MUC2 epithelial mucin in breast carcinoma.
Authors: Walsh M D , McGuckin M A , Devine P L , Hohn B G , Wright R G
Published: June 2007

AIMS--To examine the expression of the MUC2 epithelial mucin in breast carcinoma; to relate this to patient survival. METHODS--Sections from 210 breast carcinomas were stained with the anti-MUC2 core protein monoclonal antibody, 4F1, using an immunoperoxidase technique. The proportion of tumour cells positively stained and the localisation and intensity of any staining were recorded. Expression of MUC2 was compared with histological type and grade, tumour size, presence of nodal metastases, presence of oestrogen receptors, and menopausal status. The prognostic value of MUC2 expression was examined using Kaplan-Meier survival analysis. RESULTS--MUC2 mucin was detected in 19% of cases of invasive carcinoma, in 11% of cases of carcinoma in situ, where present, but very rarely in adjacent normal breast epithelium. Presence of MUC2 was significantly associated with a shorter disease free interval (p < 0.05), although the observed difference in duration of overall survival was not significant. CONCLUSIONS--The MUC2 detected in breast carcinoma may be underglycosylated or staining may represent detection of the protein core before the completion of glycosylation. The virtual absence of 4F1 reactivity in normal breast epithelium suggests that, unlike the MUC1 mucin, the MUC2 mucin is not highly expressed by these cells. The mechanism by which expression of MUC2 affects the biology of breast tumours is unclear, although expression may be a reflection of general derepression of genes during tumour progression.
Epstein-Barr virus in normal, pre-malignant, and malignant lesions of the uterine cervix.
Authors: Landers R J , O'Leary J J , Crowley M , Healy I , Annis P , Burke L , O'Brien D , Hogan J , Kealy W F , Lewis F A
Published: June 2007

AIM--To detect the presence or absence of Epstein-Barr virus (EBV) in cervical lesions ranging from normality to invasive malignancy. METHODS--Eighteen randomly selected cases of invasive squamous cell carcinomas of the uterine cervix were examined as well as 25 cases each of normal cervices and those showing cervical intra-epithelial neoplasia (CIN) I, II, and III. DNA-DNA in situ hybridisation, using a biotinylated probe to the Bam H1 "W" fragment of EBV, was carried out in addition to the polymerase chain reaction using specific primer sequences that flank a 153 base pair segment of the Bam H1 "W" region of the EBV genome and which do not cross-amplify other DNA herpes viruses. Positive control material included paraffin wax embedded P3 HR1 lymphoblastoid cells (containing high copy numbers of EBV) and two nasopharyngeal carcinomas positive for EBV. RESULTS--Neither normal nor CIN I tissue was positive. Eight per cent of CIN II tissue was positive; 8% of CIN III, and 43% of carcinomas were positive for EBV. CONCLUSION--The study shows that the virus is present in some cases of cervical carcinoma and to a lesser degree in some premalignant lesions of the cervix, but the exact association between it and cervical oncogenesis, be it causative or incidental, remains to be determined.
Detection of immunoglobulin heavy chain gene rearrangements in Hodgkin's disease using PCR.
Authors: Angel C A , Pringle J H , Primrose L , Lauder I
Published: June 2007

AIM--To detect clonal rearrangements of the immunoglobulin (Ig) heavy chain gene in Hodgkin's disease tissue using the polymerase chain reaction (PCR). METHODS--DNA extracted from 36 samples of Hodgkin's disease was analysed using PCR and primers from conserved sequences in the variable (VH) and joining (JH) regions. RESULTS--Clonal rearrangement was detected only in one case. Evidence of clonal immunoglobulin gene rearrangement had been detected previously in this case using conventional Southern blot analysis. CONCLUSIONS--The sensitivity of the two techniques is equivalent and clonal Ig heavy chain gene rearrangements are rare in Hodgkin's disease.
Nucleolar organiser regions and survival in patients with non-Hodgkin's lymphomas classified by the working formulation.
Authors: Jakić-Razumović J , Tentor D , Petrovecki M , Radman I
Published: June 2007

AIMS--To correlate the numbers of silver staining nuclear organiser regions (AgNORs) in non-Hodgkin's lymphoma classified by the working formulation with survival, the first complete remission, and the length of remission. METHODS--Sixty one patients were included in the study. Paraffin wax sections were stained using silver solution to visualise nucleolar organiser regions. The AgNORs were counted in 150 nuclei of each specimen. Data were examined using the Kruskal-Wallis test, multivariate discriminant analysis, and Cox's regression test. Curves were calculated by the method of Kaplan and Meier. RESULTS--Most patients who were alive had low grade lymphoma (p < 0.01). The first complete remission was obtained more frequently in the low and intermediate grade groups (p < 0.05). The longest survival was found in the low grade group (p < 0.001). The mean number of AgNORs differed significantly in all three groups (p < 0.001). This was also true for area of nuclei (p < 0.001) and length of remission (p < 0.05). In a multivariate analysis the numbers of AgNORs were highly predictive for survival, remission, and the length of remission. CONCLUSIONS--The numbers of AgNORs correlated with survival, remission, and the length of remission in patients with non-Hodgkin's lymphoma.
Prognostic importance of DNA flow cytometric variables in rhabdomyosarcomas.
Authors: Wijnaendts L C , van der Linden J C , van Diest P , van Unnik A J , Delemarre J F , Voûte P A , Meijer C J
Published: June 2007

AIM--To determine whether DNA ploidy patterns and S phase fraction offer prognostic information in patients with rhabdomyosarcoma (RMS). METHODS--DNA flow cytometry was performed on formalin fixed, paraffin wax embedded samples from primary tumours, and metastatic deposits or recurrences in 70 patients. DNA histogram analysis was done using a semi-automated cell cycle analysis program. RESULTS--Of the 70 primary tumours, 23 were DNA diploid, 32 DNA aneuploid, eight DNA multiploid, and seven DNA tetraploid. The prognosis for DNA aneuploid patterns was favourable, intermediate within the group of DNA tetraploid tumours and poor among patients with DNA diploid and DNA multiploid tumours (p = 0.009). In multivariate analysis (Cox regression model) DNA ploidy was an important independent prognostic factor, along with TNM stage, localisation, and histopathological classification. Ten out of 32 patients with a high S phase fraction (> 15%) with primary RMS achieved long term survival in contrast to 20 out of 29 patients with a low S phase fraction (< or = 15%) (p = 0.008). In 24 cases the DNA ploidy of cases of relapse was analysed. Of the 15 cases, in which stem line changes had occurred, 13 died of disease. No stem line changes were noted in nine cases and in this group four patients died of disease (p = 0.02). CONCLUSIONS--Assessment of DNA ploidy and S phase fraction in primary RMS and evaluation of stem line changes in cases of relapse are important variables in predicting prognosis.
T cell lymphoid aggregates in bone marrow in idiopathic hypereosinophilic syndrome.
Authors: Metz J , McGrath K M , Savoia H F , Begley C G , Chetty R
Published: June 2007

Idiopathic hypereosinophilic syndrome (HES) comprises a heterogeneous group of disorders characterised by prolonged eosinophilia with no obvious cause. A patient with longstanding HES is reported in whom unusual non-neoplastic peritrabecular lymphoid aggregates were present in the bone marrow, a hitherto undescribed association, as far as is known. An eosinophil colony stimulating activity was detected in the serum. The findings in this patient provide further evidence for an important role for eosinophil colony stimulating activity interleukin-5 mediated T lymphocyte control of eosinophil production in the pathogenesis of the HES.
Hypochromic macrocytes: are they reticulocytes?
Authors: Bain B J , Cavill I A
Published: June 2007

Automated full blood counters measuring the size and haemoglobin concentration of individual red cells by high and low angle light scatter can provide estimates of the percentage of cells which are abnormal in either or both of these variables. The hypothesis that an increase in the number of large cells with a reduced haemoglobin content ("hypochromic macrocytes") was indicative of a reticulocytosis was investigated. A correlation was shown between the percentage of hypochromic macrocytes and the reticulocyte count. This correlation was too weak to allow the actual reticulocyte percentage to be predicted from the percentage of hypochromic macrocytes. An increased percentage of hypochromic macrocytes, however, often indicates an increased reticulocyte count and thus serves to chart the haematologist to this possibility.
PCR analysis of the upstream regulatory region of human papillomavirus genomes in cervical intraepithelial neoplasia and cervical carcinoma.
Authors: Donaldson Y K , Arends M J , Duvall E , Bird C C
Published: June 2007

AIMS--To test whether human papillomavirus (HPV) variants with large scale sequence alterations to the upstream regulatory region are present in cervical intraepithelial neoplasias (CIN) and cervical carcinomas. METHODS--New PCR based assays were designed specifically to detect large scale insertion/deletion alterations in the upstream regulatory region of HPV 16 and 18. The assays were applied to 24 cases of CIN and 34 cases of cervical carcinoma previously shown to contain these two high risk HPV types. RESULTS--No large scale sequence alterations were found in any of the HPV containing CIN or carcinomas. CONCLUSIONS--These negative findings suggest that major upstream regulatory region variants of HPV 16 and 18 do not contribute to most cervical neoplasms.
Malignant transformation of osteoblastoma: study using image analysis microdensitometry.
Authors: Grace J , McCarthy S , Stankovic R , Marsden W
Published: June 2007

AIM--To determine if the malignant transformation, as perceived histologically, in a case of osteoblastoma from the right femur, was also expressed as a quantitative change in nuclear DNA during tumour progression over five months. METHODS--Nuclear DNA microdensitometry by computer image analysis was used to acquire relative DNA distribution patterns. Tissue had been removed on four separate occasions from a lesion in the right femur of an 18 year old man. Retrospective DNA analysis was performed on formalin fixed, paraffin wax-embedded tissue. RESULTS--The DNA profile of the initial biopsy specimen, which was histologically diagnosed as osteoblastoma, was euploid with a near diploid (2c) modal DNA. The second biopsy specimen taken one month later also resembled osteoblastoma but showed an aneuploid DNA profile with a diploid modal DNA and some nuclei with ploidy greater than 5c. The third biopsy specimen taken four months later showed histological evidence of osteosarcoma and a near pentaploid (5c) modal DNA with large number of nuclei exceeding 5c. CONCLUSIONS--DNA microdensitometry confirmed the initial and final diagnosis. The technique also seems to be capable of detecting aneuploidy before malignancy is morphologically evident.
Iron overload in multiply transfused patients who are HIV seropositive.
Authors: Goldin R D , Wilkins M , Dourakis S , Parkin J , Lindley R
Published: June 2007

AIMS--To assess histologically the amount of iron deposited in liver biopsy specimens from HIV positive patients; and to perform estimations of liver iron on tissue from patients with an increase in parenchymal stainable iron. To correlate the amount of blood transfused and the degree of iron overload. METHODS--Liver biopsy specimens (n = 120) from 109 HIV positive patients, 74 of whom had AIDS, were examined retrospectively and the amount of iron, as visualised with Perls's stain, was graded. Fibrosis was assessed using connective tissue stains. Estimations of liver iron were performed on tissue retrieved from paraffin wax blocks in cases with histological grade 3 or 4 iron overload. The amount of blood transfused before liver biopsy was determined from the notes for each patient. RESULTS--Fifteen of the 120 liver biopsy specimens had significantly increased amounts of iron in their hepatocytes, as assessed histologically, and this was confirmed in seven cases by measurement of liver iron. There was a close correlation between the amount of blood transfused and the degree of iron overload. In the initial biopsy specimens only one case showed portal tract expansion. Three of the five patients who had repeat biopsies, however, showed progressive fibrosis. CONCLUSION--Multiply transfused HIV positive patients may develop clinically important iron overload and are at risk of developing progressive fibrosis. Superimposed liver disease, especially viral hepatitis, in these high risk patients may exacerbate the effects of the iron overload.
Development of an internal quality assessment scheme in a clinical bacteriology laboratory.
Authors: Constantine C E , Amphlett M , Farrington M , Brown D F , Messer S , Rampling A , Warren R E
Published: June 2007

AIM--To develop an internal quality assessment (IQA) scheme in a clinical bacteriology laboratory. METHODS--Over 24 months, 1230 diagnostic specimens, representing 0.42% of laboratory workload, were anonymised and resubmitted for analysis. Six hundred and twenty one (48.7%) of these gave positive culture results; 44 fecal and upper respiratory specimens were "spiked" (artificially inoculated) to increase the proportion of positive samples. RESULTS--Discrepancies between IQA and clinical sample results occurred in 188 cases (14.8%): 76.6% of these were in culture results, 13.3% in microscopy performance, and 10.1% in clerical recording. The culture discrepancy rate for each positive sample was lowest for wound (17.5%) and urine (18.1%) specimens, and highest for faeces (34.9%) and upper respiratory (37.7%) samples. Discrepancies in several areas responded to staff training and improvement in technical methods. CONCLUSIONS--An IQA programme of this type assesses the reproducibility of tests within a diagnostic laboratory when analysing common specimen types and organisms. It permits blind assessment of many areas of diagnostic work that are not readily amenable to other quality assurance methods, and it raises the awareness of all staff to the importance of quality in every aspect of specimen and data processing.
Antimicrobial activity of cytotoxic drugs may influence isolation of bacteria and fungi from blood cultures.
Authors: Peiris V , Oppenheim B A
Published: June 2007

The potential antimicrobial activity of cyclophosphamide, vincristine, and adriamycin against Gram positive and negative bacteria and Candida albicans was examined. The time taken for different microbial inocula to turn a simulated blood culture positive in the presence of different concentrations of these drugs was measured. Doxorubicin retarded the growth of Staphylococcus aureus, Staphylococcus epidermidis, and Streptococcus sanguis in a concentration dependent manner. Cyclophosphamide and vincristine showed minimal antimicrobial activity. Escherichia coli and Pseudomonas aeruginosa were unaffected by any of the drugs. An inoculum dependent effect was seen with some combinations of microbial inocula and cytotoxic drug concentrations.
Inaccurate haemoglobin estimation in Waldenström's macroglobulinaemia: unusual reaction with monomeric IgM paraprotein.
Authors: Goodrick M J , Boon R J , Bishop R J , Copplestone J A , Prentice A G
Published: June 2007

Automated blood counts from a patient with Waldenström's macroglobulinaemia repeatedly failed critical limit standards set for mean cell haemoglobin concentration and mean cell haemoglobin. Haemoglobin estimation was higher than that suggested by clinical examination, symptoms, and the spun haematocrit. This was found to be due to an interaction between the Coulter lysing agent and monomeric IgM paraprotein in the patient's plasma, creating a precipitate which was optically dense at 525 nm.