bluuebunny/crossref_metadata_2025 huggingface dataset:
In situ hybridisation with digoxigenin-labelled DNA probes for detection of viral genomes.
Authors: Furuta Y , Shinohara T , Sano K , Meguro M , Nagashima K
Published: June 2007

The applicability of a recently developed non-radioactive DNA labelling and detection method, which uses the digoxigenin (DIG) enzyme linked immunosorbent assay (ELISA) system, for the detection of viral infections in pathology specimens by in situ hybridisation, was examined. Its efficacy was compared with that of biotin and radioisotope labelling methods. Three cases of progressive multifocal leucoencephalopathy, two of verruca vulgaris, and seven cases of laryngeal papilloma were studied. The sensitivity of the DIG labelled probe was almost the same as that of a 35S-labelled probe in the dot-blot hybridisation test. Using in situ hybridisation with 35S-labelled and DIG labelled probes, the levels of the hybridised signals detected were similar. The biotin labelled probe was less sensitive, particularly in the cases of laryngeal papilloma. The DIG labelling and detection method was highly sensitive and applicable to the detection of viral infection by ISH, and is preferable to a radiolabelled probe, especially when in situ hybridisation is done in the pathology laboratory.
Immunocytochemical analysis of lymph node aspirates in patients with human immunodeficiency virus infection.
Authors: Oertel J , Oertel B , Lobeck H , Huhn D
Published: June 2007

Thirty four patients positive for human immunodeficiency virus (HIV) who had lymphadenopathy were investigated using fine needle aspiration. Cytological analysis included immunocytochemical investigation with the alkaline phosphatase-antialkaline phosphatase (APAAP) method. All patients had confirmation of cytological diagnosis by lymph node biopsy. Fifteen aspirates with follicular hyperplasia were evaluated. Eleven patients showed B cell predominance. The B cell population did not show light chain restriction. Ten patients with B cell non-Hodgkin's lymphoma (five with Burkitt's lymphoma and five with B cell immunoblastic lymphoma) were investigated. Nine out of 10 cases were monoclonal with respect to their light chain determinants; only one case with Burkitt's lymphoma with partial lymph node metastasis did not show light chain restriction. The cytological diagnosis included two mycobacterial infections and four cystic lesions. Histological investigation was necessary to diagnose the extent of lymph node disease caused by Kaposi's sarcoma. These findings indicate that the immunocytological investigation of lymph node aspirates is useful for evaluating lymphadenopathy in HIV positive patients.
Use of latex agglutination technique for detecting Legionella pneumophila (serogroup 1) antibodies.
Authors: Holliday M G
Published: June 2007

Following the outbreak of Legionnaires' disease in Stafford in 1985, 500 serum samples were submitted to the indirect immunofluorescence antibody test and a latex agglutination. Latex agglutination using ultrasonically disrupted Legionella pneumophila antigens coupled to latex particles, proved a rapid, simple method for detecting circulating antibodies to L pneumophila in a one minute slide latex agglutination test. There was good correlation with the indirect immunofluorescence antibody test (IFAT), and the specificity and sensitivity with respect to a diagnostic result were 98.3% and 97.6%, respectively, using a series of well characterised sera. The latex agglutination test seems well suited as a screening test for presumptive cases of Legionnaires' disease; the latex reagent is easy to prepare and seems to remain stable at 4 degrees C for up to six months.
Immunoblot technique to visualise serum pepsinogen A isozymogen patterns.
Authors: Zwiers A , Toonstra C , Pals G , Donker A J , Meuwissen S G , ten Kate R W
Published: June 2007

Pepsinogen A (PGA) isozymogen patterns in urine and gastric mucosa can be visualised in non-denatured polyacrylamide gel electrophoresis by showing proteolytic activity after the conversion of pepsinogen into pepsin by acid. This method is not suitable for visualising PGA patterns in serum due to low PGA concentrations. To obtain a more sensitive visualisation method an immunoblotting technique was developed. PGA isozymogen patterns from urine and sonified gastric mucosa specimens obtained by immunoblotting were identical with those obtained by activity staining. The immunostaining method was at least 50 times more sensitive. PGA isozymogen patterns could be visualised in serum. Preliminary results suggest that the PGA patterns in serum and gastric mucosa are identical. As an association has been found between the genetically determined PGA isozymogen patterns in gastric mucosa and gastric malignancies in man, immunoblotting of PGA isozymogens in serum may provide a screening tool for subjects at risk of malignant gastric disease.
Immunohistochemical demonstration of pancreatic secretory trypsin inhibitor in normal and neoplastic colonic mucosa.
Authors: Bohe H , Bohe M , Lindström C , Ohlsson K
Published: June 2007

Specimens of normal and neoplastic colonic mucosa from 52 patients were analysed by immunohistochemistry using a monospecific polyclonal antiserum against human pancreatic secretory trypsin inhibitor (PSTI). In normal colonic mucosa PSTI was found in the goblet cells in the basal parts of the crypts. In adenomas of tubular, villous, and tubulo-villous types PSTI was also found in the upper parts of the polyps, usually occurring in the regeneration zone. There was a more intense staining reaction in polyps with increased atypia. Carcinomas of different types and of various grades of differentiation and of in situ type did not contain PSTI. These findings indicate that PSTI could be a marker for adenomatous rather than carcinomatous epithelium in the colon. Furthermore, the absence of the inhibitor in malignant cells might facilitate tissue invasion by malignant cells because of deficient protease inhibition.
Serum alanine transaminase (ALT) reference ranges estimated from blood donors.
Authors: Goldie D J , McConnell A A
Published: June 2007

It has been suggested that an increase in serum alanine transaminase (ALT) activity in blood donors may identify infection with non-A, non-B hepatitis. To facilitate identification of such donors, the reference range for ALT was measured on a Technicon SMAC 1 Analyser, using serum from 364 blood donors and 567 plasmapheresis donors. The distribution of ALT activities displayed a positive skewness, and so both logarithmic transformation and subsequent calculation of mean and standard deviation as well as non-parametric analysis were used to obtain best estimates of reference ranges for men and women, 5-65 IU/1 and 5-35 IU/1, respectively. ALT activities were found to be higher in plasmapheresis donors than in normal blood donors of both sexes, and it is postulated that this difference may be related to the increased frequency of donation in the former group.
Bone marrow stromal cell changes in haematological malignancies.
Authors: Dilly S A , Jagger C J
Published: June 2007

Stromal cell numbers from subjects with no haematological disease and those with acute myeloid leukaemia (AML), chronic granulocytic leukaemia (CGL), acute lymphatic leukaemia (ALL) and non-Hodgkin's lymphoma (NHL) were compared to determine their role in malignancy. Frozen sections of trephine biopsy specimens from iliac crests were stained for endogenous alkaline phosphatase activity, endogenous acid phosphatase activity, and, using immunocytochemical methods, for endothelial cells (anti-factor-VIII related antigen) and macrophages and related cells (EBM/11). In granulocytic malignancies, whether acute or chronic, alkaline phosphatase positive reticulum cells (AL-RC) and vascular endothelial cells were generally increased. In lymphoid malignancies, the numbers of AL-RC were generally reduced. Numbers of vascular endothelial cells seemed to be normal in ALL but reduced in foci of NHL. Macrophages are numerous in normal marrow, and their numbers seemed to be normal in granulocytic lesions but were more variable and sometimes reduced in ALL and NHL. Lymphoid malignancies, therefore, have a destructive effect on some stromal elements; granulocytic malignancies are associated with normal or increased numbers of stromal cells. A possible consequence of depleted stromal cells might be slower reconstitution of normal haemopoiesis after treatment. The large numbers in granulocytic malignancies raises the possibility of synergistic stimulation between stromal and neoplastic cells.
Non-specific reactions in enzyme linked immunosorbent assays for serum antibody to entamoeba histolytica and Giardia lamblia in non-endemic areas.
Authors: Shetty N P , Raj I S , Macaden R S
Published: June 2007

Serum samples from 20 Indian children with diarrhoea were compared with those from 20 children resident in the United Kingdom who had been diagnosed as having ulcerative colitis, or Crohn's disease, or indeterminate colitis using enzyme linked immunosorbent assays specific for Entamoeba histolytica and Giardia lamblia. More than 50% of the United Kingdom patients had high IgG responses in ELISAs for E histolytica and G lamblia. A confirmatory ELISA showed that the British sera reacted specifically to bovine serum proteins rather than to protozoal antigens. Prior incubation of sera with 5% bovine serum prohibited this reaction. Bovine serum is an integral part of the crude soluble antigen used in most ELISAs for E histolytica and G lamblia and needs to be replaced with purified antigen preparations. The British sera also reacted to other commonly used blocking agents such as bovine serum albumin, casein, and normal sheep serum. These reactions were attributed to uptake of dietary antigens or an enhanced immunological response to these antigens in patients with inflammatory bowel disease.
Abnormal fucosylation of-ileal mucus in cystic fibrosis: II. A histochemical study using monoclonal antibodies to fucosyl oligosaccharides.
Authors: King A , McLeish M , Thiru S
Published: June 2007

Abnormal fucosylation of cystic fibrosis mucin was previously shown using peroxidase conjugated lectins on ileal tissue sections. These abnormally fucosylated glycoproteins were investigated further using monoclonal antibodies to fucosyl oligosaccharides based on type 1 and type 2 blood group precursor chains. The results of this study, using monoclonal antibodies to blood group glycoproteins in cystic fibrosis, were negative, yet abnormal fucosylation had been found using lectin histochemistry. Using monoclonal antibodies, lectins, and appropriate enzymes, such as glycosyl hydrolases, it should be possible to delineate further the abnormality found in glycoproteins in cystic fibrosis on appropriately fixed ileal sections, obtained from infants at term presenting with meconium ileus.
Potential role of IgG avidity for diagnosing toxoplasmosis.
Authors: Joynson D H , Payne R A , Rawal B K
Published: June 2007

Sera from 20 cases of toxoplasmic lymphadenopathy were examined by an enzyme linked immunosorbent assay toxoplasma IgG avidity (ELISA) at two laboratories. The results obtained were largely in agreement and showed that sera from patients with acute infection had low avidity IgG (30% or less), whereas sera from patients with chronic infection had high avidity IgG (40% or more). It is suggested that this type of assay could have a useful complementary role in antenatal testing for toxoplasmosis.
Gastric epithelium in the duodenum: its association with Helicobacter pylori and inflammation.
Authors: Wyatt J I , Rathbone B J , Sobala G M , Shallcross T , Heatley R V , Axon A T , Dixon M F
Published: June 2007

Duodenal biopsy specimens from 471 adults and 47 children were examined to determine the prevalence and distribution of gastric epithelium in the duodenal bulb in relation to age, gender, gastroduodenal inflammation, smoking, alcohol and consumption of nonsteroidal anti-inflammatory drugs (NSAID). Gastric metaplasia was present in the anterior wall duodenal biopsy specimen in 31%, was significantly less common in patients under 17 than in adults, and was more common in males than females. In sixty two adults who underwent multiple radial duodenal biopsy gastric metaplasia was randomly distributed around the duodenal circumference; sixty three per cent of the patients with gastric metaplasia found on multiple biopsy were detected by just the anterior biopsy. Gastric metaplasia was not obviously associated with alcohol, cigarette, or NSAID consumption. While the presence of gastric metaplasia was associated with adulthood, male sex, and low fasting gastric juice pH, its extent was associated with active duodenitis and Helicobacter-associated gastritis. On logistic regression, gastric metaplasia in the duodenum and gastric Helicobacter pylori were independent predictors of active duodenitis, but were not significantly associated with inactive duodenal inflammation. H pylori was observed in duodenal biopsy specimens from 32 patients, all with active duodenitis; bacteria were present only on foci of gastric metaplasia, and were more likely to be seen when the metaplasia was extensive. It is proposed that inflammatory injury to the duodenal mucosa by H pylori may stimulate the development of further gastric metaplasia, and that the area of duodenum susceptible to colonisation with H pylori may therefore increase progressively until mucosal integrity is compromised and ulceration supervenes.
Labelling of cells of the mononuclear phagocyte system in routinely processed archival biopsy specimens with monoclonal antibody EBM/11.
Authors: Greywoode G I , McCarthy S P , McGee J O
Published: June 2007

When first reported, EBM/11 reacted with mononuclear phagocyte system cells only in fresh frozen sections, but it has been shown to have similar cellular specificity in routine formalin fixed, paraffin wax embedded tissue. This was achieved by limited proteolysis with protease XIV before immunocytochemical staining. In archival biopsy specimens EBM/11 produced granular cytoplasmic staining of alveolar macrophages, Kupffer cells, tingible body macrophages and sinus histiocytes, cells of splenic cords, cortical and medullary macrophages of thymus; blood monocytes, peritoneal and mesothelial macrophages; bone marrow mononuclear cells, megakaryocytes and osteoclasts; lamina propria macrophages in the gastrointestinal tract, and connective tissue cells (presumptive macrophages) of thyroid, gall bladder, skin, pancreas, ovary, myometrium, endometrium, cervix, kidney, prostate, placenta, myocardium and breast. Unlike other anti-macrophage antibodies, EBM/11 did not react with granulocytes, lymphocytes, plasma cells, platelets, endothelial and epithelial cells in paraffin wax sections. It did not label skin Langerhans' cells, microglial cells, and interdigitating reticulum cells (as in frozen sections). This study opens a new area for the specific identification by EBM/11 of mononuclear phagocyte system cells in archival biopsy specimens. It also raises the possibility that some monoclonal antibodies, believed to be reactive only in frozen sections, may react in archival tissue after limited proteolysis with an appropriate enzyme.
Monitoring rejection after heart transplantation: cytoimmunological monitoring on blood cells and quantitative birefringence measurements on endomyocardial biopsy specimens.
Authors: Wijngaard P L , Gimpel J A , Schuurman H J , van der Meulen A , Gmelig Meyling F H , Jambroes G
Published: June 2007

Cytoimmunological monitoring and quantitative birefringence measurements were used as potential aids in diagnosing acute rejection after heart transplantation instead of histopathological assessment of the endomyocardial biopsy specimen alone. Cytoimmunological monitoring was based on morphological inspection and quantitation of mononuclear cells, particularly activated lymphoid cells. Quantitative birefringence measurements comprise a variable for myocyte contractile function. Its read out is the ratio of the degree of birefringence before contraction to that after. Cytoimmunological monitoring indicated significantly higher concentrations of activated lymphocytes in moderate or severe acute rejection, and quantitative birefringence measurements indicated decreased myocyte function during severe and resolved or resolving rejection. Cytoimmunological monitoring and quantitative birefringence measurements were diagnostically most useful in terms of sensitivity, specificity, and predictive value, when only data gathered before the first episode of acute rejection were considered. For cytoimmunological monitoring, diagnostic relevance was optimal when the data were expressed as relative proportions of activated lymphocytes. The quantitative birefringence measurements correlated best with analysis of the endomyocardial biopsy specimen when a cut off value of 1.25 was used. When both methods for diagnosing acute rejection were analysed together, no improvement in sensitivity (value 0.44) was found, but the specificity increased to 0.98 and the predictive value to about 0.80. It is concluded that cytoimmunological monitoring is a useful, non-invasive additional method for diagnosing the first period of acute rejection after heart transplantation and that quantitative birefringence measurements give valuable information on the extent of myocyte damage.
Nucleolar organiser regions in kidney tumours and xanthogranulomatous pyelonephritis.
Authors: Bryan R L , Crocker J , Farr A
Published: June 2007

The AgNOR argyrophil technique was applied to clear cut examples of renal adenoma (n = 7), clear cell carcinoma (n = 9), and xanthogranulomatous pyelonephritis (n = 5). There was no significant difference between mean AgNOR counts per cell in adenomas and carcinomas, supporting the idea that there is a continuous spectrum of kidney tumours. Xanthogranulomatous pyelonephritis and clear cell carcinoma showed significant differences in mean AgNOR counts per cell, suggesting that the technique may be of use in distinguishing between these conditions, although further cases may need to be examined to confirm this.
Prognostic value of nucleolar morphometric variables in cytological breast cancer specimens.
Authors: van Diest P J , Mouriquand J , Schipper N W , Baak J P
Published: June 2007

In a retrospective study on cytological specimens from 86 patients with histologically confirmed invasive breast cancer, the prognostic value of nucleolar morphometric variables was studied and compared with nuclear variables. One hundred nuclei and their nucleoli on each slide were measured with a graphic tablet system at a total magnification of 2800 times using a stratified selection method. The number of nucleoli per 100 nuclei was also noted. Analysis of Kaplan-Meier univariate recurrence free survival curves showed significant differences for eight nuclear features, nine nucleolar features, and three combined nuclear and nucleolar variables. The total number of nucleoli per 100 nuclei was the best single prognostic variable. Multivariate survival analysis (Cox regression model) showed that no other features provided additional prognostic information beyond that given by the total number of nucleoli. It is concluded that nucleolar morphometric variables assessed in cytological preparations have prognostic value in breast cancer, and the results of this study suggest that their prognostic value may exceed that of nuclear variables.
Computer assisted diagnosis of fine needle aspirate of the breast.
Authors: Heathfield H A , Kirkham N , Ellis I O , Winstanley G
Published: June 2007

The development of the National Breast Screening Programme has created a demand for the widespread availability of fine needle aspiration cytology services. To meet this demand there must be a rapid increase in the number of pathologists and laboratories able to offer this service. In turn there is a need for improved training methods. The technique of fine needle aspiration cytology is not inherently complicated. The number of possible conclusions is essentially limited to four: unsatisfactory, benign, suspicious and malignant. A computer based expert system, designed to assist pathologists in the diagnosis of fine needle aspirates of the breast, has been developed. The system prompts pathologists to categorize a number of variables in the aspirate including nuclear and cytoplasmic features, and the degree of cellular cohesion, and uses these data to reason about possible conclusions. The final diagnosis is displayed with a detailed explanation listing the factors supporting it. Initial trials with this system have been encouraging and it is envisaged that this system will be of value both in training and as an aid to routine diagnosis.
Idiopathic portal hypertension associated with cytotoxic drugs.
Authors: Shepherd P , Harrison D J
Published: June 2007

Four patients developed clinically important portal hypertension with histological features of idiopathic portal hypertension while they were receiving cytotoxic drugs for chronic myeloid leukaemia and Hodgkin's disease. Mild sclerosis of some small portal triads was the only abnormality seen at light microscopical examination in three of the four cases. In the remaining case light microscopical findings seemed to be normal. Two cases examined by electron microscopy showed perisinusoidal fibrosis; in one case this was the only abnormality detected. There is an association between idiopathic portal hypertension and the use of chemotherapeutic agents, particularly thioguanine. Adequate histological examination of liver tissue, including electron microscopic studies, is recommended for patients who develop hepatic problems while receiving cytotoxic treatment to elucidate this problem.
Methylation of DNA in megaloblastic anaemia.
Authors: Perry J , Chanarin I , Deacon R , Lumb M
Published: June 2007

Methylation of cytosine residues in DNA samples, collected before and serially after cobalamin treatment from patients with cobalamin deficiency, was studied using restriction endonucleases Hpa II and Msp I and an epsilon globin gene probe. There was no evidence of hypomethylation in any of the samples. It was concluded that although hypomethylation of metabolites such as choline occurs, that of DNA is preserved in megaloblastic anaemia.
Demonstration of Epstein-Barr virus in primary brain lymphoma by in situ DNA hybridisation in paraffin wax embedded tissue.
Authors: Murphy J K , Young L S , Bevan I S , Lewis F A , Dockey D , Ironside J W , O'Brien C J , Wells M
Published: June 2007

Tumour tissue from 29 patients with primary brain lymphoma was reviewed to determine if there was an aetiological association between Epstein-Barr virus and polyclonal and monoclonal lymphoproliferations. The morphology and immunophenotype in 24 patients for whom paraffin wax embedded tissue was available were studied. A high grade pleomorphic tumour morphology with plasmacytoid features was seen in 13 tumours. Because of the large number of pleomorphic lymphomas, all tumours were examined for the presence of the Epstein-Barr virus genome using in situ DNA hybridisation. A panel of three biotinylated probes to different sequences in the Epstein-Barr virus genome was used. Positive hybridisation with one or more probes was shown in tumours from 11 patients. The remaining tumours gave no hybridisation signal. There was no correlation between positive hybridisation and morphological subtype or clinical outcome.
Lead poisoning: clinical, biochemical, and haematological aspects of a recent outbreak.
Authors: Pagliuca A , Mufti G J , Baldwin D , Lestas A N , Wallis R M , Bellingham A J
Published: June 2007

The clinical, biochemical, and haematological aspects of a recent outbreak of lead poisoning, in which exposure was related to the oxyacetylene cutting of red lead painted ironwork, were investigated. Initial suspicion was raised when a blood film showed punctate basophilia which remains a simple and useful method of picking up lead toxicity. Estimations of blood lead concentration and conventional laboratory data confirmed the diagnosis. Although there was prominent punctate basophilia, spectrophotometric analysis showed only negligible accumulation of pyrimidine-5'-nucleotides despite severe suppression of pyrimidine-5'-nucleotidase activity. The pattern of the red cell glycolytic intermediates, investigated for the first time, suggested that lead may also affect glycolysis at the hexokinase step. Once the diagnosis was made intravenous chelation treatment was begun with a rapid improvement in symptoms. Long term follow up is required to assess any sequelae of intoxication. These cases emphasise the classic features of lead poisoning, and despite the currently available diagnostic tests, lead intoxication may still go unrecognised unless a thorough occupational history is taken.
Reflux gastritis in the intact stomach.
Authors: Sobala G M , King R F , Axon A T , Dixon M F
Published: June 2007

Gastric biopsy specimens from patients who have undergone gastric surgery frequently exhibit foveolar hyperplasia, oedema, vasodilatation and congestion, and a paucity of inflammatory cells as consequences of entero-gastric reflux. Similar, albeit generally milder, changes were found in 47 of 316 (15%) non-surgical patients undergoing endoscopy for dyspeptic symptoms. To relate these changes to bile reflux or other potential gastric irritants the total bile acid concentration was measured in samples of fasting gastric juice, and the use of a symptom questionnaire ascertained the patients' cigarette consumption, use of non-steroidal anti-inflammatory drugs (NSAIDs), and alcohol intake. When patients with reflux gastritis were compared with normal controls (n = 91), significant increases in associated peptic ulceration and NSAID use were found in the group with reflux, but no increases in bile acid concentrations. Indeed, only one patient had evidence of duodenogastric reflux. It is concluded that most cases of "reflux gastritis" in the intact stomach are not due to reflux of bile. Our findings indicate an important pathogenic role for long term NSAID use, in what might be usefully termed type C or "chemical" gastritis.
Serum bactericidal resistance of faecal Escherichia coli and bactericidal competence of serum from patients with ulcerative colitis.
Authors: Burke D A , Clayden S A , Axon A T
Published: June 2007

A microtitre method was developed to screen Escherichia coli from 48 patients with ulcerative colitis and 25 controls for serum resistance. Bactericidal resistance was indicated by a change in colour of indicator due to acid production by viable organisms and quantitated by a change in absorbance. The method clearly differentiated between organisms confirmed as resistant or sensitive by conventional techniques. Twenty four (50%) disease and 14 (56%) control E coli specimens showed serum resistance. Bactericidal competence of sera from patients with ulcerative colitis was assessed by incubating sensitive E coli with sera from 10 patients with ulcerative colitis and pooled normal serum. All sera effectively reduced viable counts to less than 6% of original inoculum. This study shows that serum samples from patients with ulcerative colitis are bactericidally competent and that there is no increase in the number of serum resistant E coli in patients with ulcerative colitis.
Proliferation in non-Hodgkin's lymphoma: a comparison of Ki-67 staining on fine needle aspiration and cryostat sections.
Authors: Brown D C , Gatter K C , Mason D Y
Published: June 2007

Assessment of the growth fraction of non-Hodgkin's lymphomas may provide useful additional prognostic information to that obtained with conventional histological criteria. The monoclonal antibody Ki-67 has been reported to provide such information immunocytochemically in tissue biopsy specimens from lymphoma as well as other tumours. This study was undertaken to assess whether this approach could be extended to fine needle aspiration (FNA) biopsy specimens which are becoming increasingly important in the diagnosis of lymphoma. In 21 cases of non-Hodgkin's lymphoma the rate of tumour proliferation estimated by Ki-67 immunostaining of FNA material, obtained from surgically removed specimens, was compared with that obtained on tissue biopsy. The correlation between both preparations was excellent, indicating that FNA biopsy material is suitable for the immunocytochemical assessment of the growth fraction of non-Hodgkin's lymphoma.
Immunohistochemical detection of neuroblastoma in frozen sections of bone marrow trephine biopsy specimens.
Authors: Reid M M , Malcolm A J , McGuckin A G
Published: June 2007

Frozen sections from bone marrow trephine biopsy specimens from children with disseminated neuroblastoma were stained using the monoclonal antibody UJ13A. An immunoalkaline phosphatase technique was the preferred staining method. Acceptable histological detail was obtained from this material and deposits of tumour cells detected. Some apparently fibrous tissue was also stained by this antibody. The results show that this immunohistological approach is feasible and provide encouragement for its addition to the range of investigations currently available for assessing the marrows of children with this disease.
Evaluation and comparison of neutrophil bipolar shape formation with a migration assay.
Authors: Lord R A , Roath S
Published: June 2007

The neutrophil shape change response to a chemotactic formylpeptide was assessed. Neutrophil bipolar shape formation (BSF) was also simultaneously assessed with a Boyden chamber-based neutrophil migration assay. Both assays were precise and relatively reproducible; the average coefficient of variation for the BSF assay was 9.6% and 9.2% for the migration assay. In a blind study the BSF assay showed 100% sensitivity at detecting subjects with known abnormal neutrophil migration. Unlike the migration assay, the BSF assay does not require isolated neutrophils, reducing possible cell activation and monitoring the cell response under more physiological conditions. Small blood samples--1 ml or less compared with 20-40 ml for the migration assay--are used, and the method is technically simple. Results are available within 40 minutes, and routine (EDTA) blood samples are used. It is concluded that the BSF assay is a suitable motility screening test for both the clinical and pharmacological examination of the movement of polymorphonuclear leucocytes.
Nucleolar organiser regions: new prognostic variable in breast carcinomas.
Authors: Sivridis E , Sims B
Published: June 2007

Nucleolar organiser regions (NORs), which are important for regulating protein synthesis, were identified in 20 breast carcinomas by means of a silver (Ag) staining technique. Infiltrating neoplasms with metastases in four or more axillary lymph nodes possessed, on average, a greater number of AgNORs per cell nucleus compared with neoplasms without nodal disease, or with one to three positive lymph nodes. The size, morphology, and distribution of AgNORs within the nucleus were also different in the two study groups. Overall, these findings suggest that breast carcinomas with multiple, irregular, and widely dispersed AgNORs tend to be of high grade malignancy.
Blind comparison of Abbott and Dupont HIV antigen ELISA tests for detecting antigenaemia in asymptomatic human immunodeficiency virus antibody positive homosexual men.
Authors: Shanson D C , Brown K E , Goonesekera S , Gazzard B , Coates A R
Published: June 2007

Three hundred and ninety eight serum samples from 270 human immunodeficiency virus (HIV) antibody positive asymptomatic homosexual men were tested in the Abbott and Dupont HIV antigen ELISA tests. In the Abbott test 62 (16%) of the sera were positive, according to the manufacturer's instructions, compared with 55 (14%) in the Dupont test. Twenty six sera were positive with the Abbott test but negative with the Dupont test and 19 sera were positive only by the Dupont test. Only 36 (9%) of the sera were positive in both tests. The Abbott confirmatory neutralisation test gave excellent agreement with the initial Abbott HIV antigen ELISA test; the Dupont confirmatory test was only in agreement with the initial positive Dupont antigen ELISA test in one third of the sera tested. Although the overall sensitivity of each of the two commercial assays tested was similar, the Abbott method may be preferable for clinical purposes if confirmation of an initial ELISA positive test result by neutralisation assay is required.
Proliferation rate of colonic mucosa in normal subjects and patients with colonic neoplasms: a refined immunohistochemical method.
Authors: Welberg J W , de Vries E G , Hardonk M J , Mulder N H , Harms G , Grond J , Zwart N , Koudstaal J , de Ley L , Kleibeuker J H
Published: June 2007

An increased colonic epithelial proliferation rate and an increase of the cryptal proliferative zone are probable markers of increased susceptibility to colonic cancer. In this study an immunohistochemical method using 5-bromo-deoxyuridine (BrdUrd) to measure the proliferation rate of colonic mucosa in vitro was used. Fresh endoscopic colonic biopsy specimens were incubated with BrdUrd and then processed for immunohistochemistry using a monoclonal antibody. Essential procedures with respect to the equal distribution of nuclei stained with BrdUrd in the biopsy specimens proved to be the cutting of the specimens before incubation and the use of a microwave oven at the beginning of incubation. The use of the procedure of the running average showed that 12 length cut crypts are sufficient to determine reliably the proliferation rate, expressed as the labelling index (LI). This was determined in the biopsy specimens of 10 subjects without organic colonic disease, eight patients with adenomatous colonic polyps, and in six patients with (recent) colonic carcinoma. Mean LI in the controls was significantly lower than in patients with colonic polyps and in those with colon cancer. It is concluded that this method is promising for screening persons at risk for colon cancer and will be of great potential in performing dietary intervention studies in these subjects.
Preferential localisation of human lymphocytes bearing gamma delta T cell receptors to the red pulp of the spleen.
Authors: Bordessoule D , Gaulard P , Mason D Y
Published: June 2007

About 4% of human T cells carry antigen receptor composed of gamma and delta chains (rather than alpha and beta chains). Double immunoenzymatic staining of frozen sections of 14 samples of human spleen showed that gamma delta bearing T cells were preferentially localised in the red pulp of this organ where on average they accounted for 17% of all T cells. There was no correlation between the number of gamma delta T cells and the diagnosis, with the exception of a case of malaria in which an unusually high number (40%) of T cells were of this type. The gamma delta bearing T cells were scattered randomly through the red pulp, and double staining combined with a marker of splenic sinusoids (CD36) showed that almost all lie outside the sinusoids within the cords of the red pulp. It is suggested that the double immunoenzymatic technique could be used for further studies of the prevalence of gamma delta bearing T cells in lymphocytic infiltrates.
Bone marrow biopsy in monoclonal gammopathies: correlations between pathological findings and clinical data. The Cooperative Group for Study and Treatment of Multiple Myeloma.
Authors: Riccardi A , Ucci G , Luoni R , Castello A , Coci A , Magrini U , Ascari E
Published: June 2007

Between January 1987 and October 1989, 561 consecutive untreated patients with monoclonal gammopathy of undetermined clinical importance (MGUS) (n = 295) or with multiple myeloma (n = 266) were evaluated in a multicentre trial. Both bone marrow biopsy and aspiration (performed at different anatomical sites) were required at presentation. Bone marrow biopsy data indicated that changes in bone marrow composition from MGUS to early multiple myeloma and to advanced multiple myeloma followed a precise pattern, including an increased percentage of bone marrow plasma cells (BMPC%), a shift from plasmocytic to plasmoblastic cytology, an increase in bone marrow cellularity and fibrosis, a change in bone marrow infiltration (becoming diffuse rather than interstitial), a decrease in residual haemopoiesis and an increase in osteoclasts. In multiple myeloma the BMPC% of biopsy specimens and aspirate were closely related, although in 5% of cases the difference between the two values was greater than 20%. Some histological features were remarkably associated with each other. For example, BMPC% was higher in cases with plasmoblastic cytology, heavy fibrosis, or reduced residual haemopoiesis. Anaemia was the clinical characteristic most influenced by bone marrow histology. The BMPC% was the only histological variable which affected the greatest number of clinical and laboratory characteristics, including, besides haemoglobin concentration, erythrocyte sedimentation rate, radiographic skeletal bone disease, and serum concentrations of monoclonal component, calcium, beta 2-microglobulin and thymidine kinase activity. These data indicate that comparative bone marrow histology in monoclonal gammopathies has clinical importance.
Bacteriostatic and bactericidal actions of boric acid against bacteria and fungi commonly found in urine.
Authors: Meers P D , Chow C K
Published: June 2007

Boric acid has been used for over 20 years to preserve urine while in transit for bacteriological examination. It has been suggested that it may be toxic for some urinary pathogens. To investigate this several strains of bacteria and fungi commonly found in urine were exposed to different concentrations of boric acid in nutrient broth. Viable counts were made at the outset and at intervals for up to 24 hours at room temperature to detect bacteriostatic or bactericidal effects. At concentrations between 10 and 20 g/l boric acid was bacteriostatic or fungistatic for very nearly all the common urinary pathogens. At 10 g/l boric acid was weakly bactericidal for some strains of Acinetobacter calcoaceticus and Pseudomonas aeruginosa, though higher concentrations were bacteriostatic only. Group B streptococci varied in their response to boric acid, but for most of them 10 or 20 g/l was satisfactorily bacteriostatic. It is concluded that boric acid is rarely toxic, and when it is, the effect is usually sufficiently delayed to be of only theoretical importance.
Assessment of the one hour adrenocorticotrophic hormone test in the diagnosis of attenuated 21-hydroxylase deficiency.
Authors: Innanen V T , Vale J M
Published: June 2007

Attenuated or partial 21-hydroxylase deficiency is one of several biochemical defects in steroid metabolism that can lead to hirsutism in young women after puberty. The diagnosis is made through the exaggerated response of 17 alpha-hydroxyprogesterone (17-OH) to adrenocorticotrophic hormone (ACTH). To provide reference data 72 mild to moderately hirsute patients aged 18 to 35 were studied (over two years) with the ACTH test. Four patients with an exaggerated response were found. The mean (+/- 1SD) for zero time was 2.4 (0.96) nmol/l and for the 60 minute time was 7.2 (2.04) nmol/l. A subpopulation was found with a significantly higher baseline at 6.2 (1.3) nmol/l but a blunted 60 minute response at 8.7 (2.5) nmol/l. This is important because of the potential confusion arising from the known variability in baseline values in previously reported patients with partial 21-hydroxylase deficiency. Extending the test to 90 minutes did not show further increase in the 17-OH response to ACTH, thus confirming the validity of the 60 minutes ACTH test. The cortisol response to ACTH was also studied. One patient with presumptive partial 21-hydroxylase deficiency overlapped in cortisol response with eight of the reference population. Theoretically, a blunted cortisol response would be expected because of the postulated enzyme block, and these results suggest that other steroid enzyme defects should also be considered when an exaggerated 17-OH response to ACTH is seen.
Comparison of ELISA antigen preparations alone or in combination for serodiagnosing Helicobacter pylori infections.
Authors: Hirschl A M , Rathbone B J , Wyatt J I , Berger J , Rotter M L
Published: June 2007

The immunoglobulin G antibody response to Helicobacter pylori was assessed in 78 patients with non-ulcer dyspepsia using five different antigen preparations. All patients were endoscoped and biopsied. The H pylori state was determined histologically on at least two endoscopic biopsy specimens using a modified Giemsa stain. The ultracentrifuged cell sonicate, acid glycine extract, and 120 kilodalton protein antigens were specific in diagnosing infection (95-98%), but had only moderate sensitivity (70-84%). By mixing either of the two complex antigens with the 120 kilodalton protein, the sensitivity of the test was increased to 97% without affecting the high specificity. The combination of ultracentrifuged sonicate or acid glycine extract with the 120 kilodalton protein therefore seems to be superior to the individual antigen preparations and is particularly suitable for the serodiagnosis of H pylori infection.
Immunoalkaline phosphatase technique applied to paraffin wax embedded tissues in diagnostic renal pathology.
Authors: Jackson R , Holme E R , Phimister G M , Kennedy A , McLay A L
Published: June 2007

An indirect immunoalkaline phosphatase (IAP) technique was used to evaluate the glomerular deposition of immunoglobulins, C3, C1q and fibrinogen. In 80 renal biopsy specimens the results obtained using this technique were compared with those obtained by direct immunofluorescence to see if it could be used as a viable alternative. The IAP technique was straightforward to perform, it yielded quick results, and was highly reproducible, provided that a standardised short fixation period of two and a half hours was used. For the detection of immunoglobulin deposits, the IAP results correlated well with those of immunofluorescence. Despite poorer performance in identifying complement components and fibrinogen it could, within certain limits, provide an adequate diagnostic alternative to immunofluorescence. Each technique gave false negative results, those of immunofluorescence being related to its failure to identify mesangial deposition of IgA in two cases where its distribution seemed to be focal, and those of IAP to a failure to detect linear deposition of IgG in all three cases of anti-glomerular basement membrane disease.
Reticulocyte counting using flow cytometry.
Authors: Nobes P R , Carter A B
Published: June 2007

A flow cytometric method for the quantitation of reticulocytes was refined for routine laboratory use. Blood (2 microliters) is added to 2 ml of 0.4 microM thiazole orange in phosphate buffered saline, incubated at room temperature for 90 minutes, and analysed on a Coulter EPICS Profile flow cytometer, with gating for red cells on the basis of forward and right angled light scatter. Blood (2 microliters) is also incubated with phosphate buffered saline alone as an unstained control. The adult reference range (mean +/- 2 SD), established from 30 laboratory personnel, is 19.4-59.2 x 10(9)/l (0.2-1.6%). Comparison of this technique was made on 39 selected patient samples with visual counting of cells stained with brilliant cresyl blue. The correlation between the two methods was 0.99 with slope 0.96 and intercept 0.02. The precision of the automated technique in three subjects with reticulocyte counts of 0.12%, 1.84%, and 14.3% was 33.3%, 7.3%, and 1.4%, respectively (coefficient of variations). In three patients studied serially after intensive chemotherapy, in whom the reticulocyte count quantitated by routine visual methods approached zero (0-0.1%) for eight to 18 days, the automated counts varied between 0 and 0.5%. Flow cytometric reticulocyte counting is thus a simple and highly reliable methodology for the quantitation of normal and raised reticulocyte counts but cannot be reliably used to quantitate a subnormal level.
Evaluation of urinary antigen ELISA for diagnosing Legionella pneumophila serogroup 1 infection.
Authors: Birtles R J , Harrison T G , Samuel D , Taylor A G
Published: June 2007

The enzyme linked immunosorbent assay (ELISA) described was developed to detect a soluble antigen in the urine of patients with Legionnaires' disease caused by Legionella pneumophila serogroup 1 (L.pn 1). The assay was evaluated and showed good specificity (100%) and intra-assay reproducibility. Antigen was detected in the urine of 93 (77%) of 120 patients, overall, and in 86% of patients from whom a specimen obtained within seven days of onset of illness was available. On all but one occasion the first urine sample taken from a patient for whom a positive ELISA result was obtained, was itself positive. In one case antigen was not detected at four days but was present on the fifth day after onset of symptoms. In two patients urinary antigen was detectable as early as two days after onset of symptoms. In another the antigen persisted for at least 60 days. More than half the patients, however, had stopped producing detectable antigen within 14 days of onset of symptoms. It is therefore important that where Legionnaires' disease is suspected urine is collected as early as possible in the course of the disease.
Specimen transport audit.
Authors: Garner P , Masterton R G
Published: June 2007

A specimen transport audit was performed at a routine and reference laboratory. Over the survey period (1986-89) the percentage of specimens received and assessed as hazardous (inadequately packed, misidentified, or contaminated by leakage) fell significantly from 12.0 to 2.8%. Specimen transport audit identified technical and logistical faults associated with sample transmission. It is concluded that no type of hazard should exceed 0.5% of samples, with the total being less than 1% of specimens received. Specimen transport audit is an additional laboratory performance indicator.
Serum pepsinogen I and gastrin concentrations in children positive for Helicobacter pylori.
Authors: Oderda G , Vaira D , Dell'Olio D , Holton J , Forni M , Altare F , Ansaldi N
Published: June 2007

Serum pepsinogen I, serum gastrin concentration, and inflammatory scores were measured in a population of 71 children undergoing upper gastrointestinal endoscopy for investigation of upper abdominal pain. Forty four were initially colonised with Helicobacter pylori. The indices were measured before treatment (in 71 children), one month (in 41 children), and six months (in 21 children) after stopping treatment. Before treatment there was a significant correlation between serum pepsinogen concentration, total inflammatory score, and H pylori state, but no correlation between serum gastrin concentrations and H pylori state. Similarly, the total inflammatory score and serum pepsinogen concentrations were significantly correlated. There was no such correlation in children negative for H pylori. After treatment the inflammatory score improved in those patients in whom H pylori had been eradicated. There was also a significant fall in serum pepsinogen I and serum gastrin concentration in those patients in whom H pylori had been eradicated. These results were similar to those found six months after treatment had been stopped. These findings suggest that the serum pepsinogen I concentration could be considered a useful marker for gastritis and can be used as an index of severity of gastritis in H pylori positive subjects. The measurement of serum gastrin concentrations does not give useful information.