bluuebunny/crossref_metadata_2025 huggingface dataset:
Lamina propria eosinophils and mast cells in ulcerative colitis: comparison between Asians and Caucasians.
Authors: Benfield G F , Bryan R , Crocker J
Published: June 2007

To investigate the theory of hypersensitivity in the colonic mucosa of Asian patients with ulcerative colitis the rectal biopsy specimens of Asian and Caucasian patients presenting with colitis were selectively stained for both eosinophils and mast cells. Comparisons between ethnic groups were made as well as the correlation to the blood eosinophil count and two variables of active ulcerative colitis--the blood leucocyte count and serum orosomucoid concentration. No correlation was found in either group between circulating and tissue eosinophils and no ethnic difference was identified in the lamina propria differential cell densities. The leucocyte count and serum orosomucoid concentrations, however, were significantly higher in the Caucasians. In the Asian group a strong negative association was found between the lamina propria eosinophil density and serum orosomucoids; in the Caucasian group there was a positive association between all blood variables. It is concluded that different mechanisms occur in the two ethnic groups and in Asians these might not be reflected by the more orthodox indices of colonic inflammation.
Changes in alveolar macrophage, monocyte, and neutrophil cell profiles after smoke inhalation injury.
Authors: Riyami B M , Tree R , Kinsella J , Clark C J , Reid W H , Campbell D , Gemmell C G
Published: June 2007

Thirty two fire victims with smoke inhalation, with or without burns, and 26 control subjects had bronchoalveolar lavage performed. Cell yields and differential cell counts were assessed. All patients and controls were cigarette smokers. Patients with smoke inhalation (SI) injury generally showed higher total bronchoalveolar lavage (BAL) cell yields, and this was significant on repeat lavage from 12 patients. The increase was almost entirely due to an increase in the proportion of neutrophils in patients with smoke inhalation alone (S) and those with cutaneous burns as well as smoke inhalation (S + B). On sequential lavage of 12 patients with smoke inhalation (SI) the proportion of neutrophils had increased; this was significantly higher than on initial lavage. Using various macrophage markers, the proportions of macrophage subgroups were determined. There was an increase in UCHM1 and RFD9 positive cells in each subgroup: the increase in UCHM1 positive cells was significant in patients with burns as well as smoke inhalation, and the increase in RFD9 positive cells was significant in patients with smoke inhalation alone. Assessment of the role of such cells in the development of acute lung injury (such as adult respiratory distress syndrome) may be important in our understanding of the mechanisms entailed.
Campylobacter pylori in the upper gastrointestinal tract of patients with HIV-1 infection.
Authors: Francis N D , Logan R P , Walker M M , Polson R J , Boylston A W , Pinching A J , Harris J R , Baron J H
Published: June 2007

Fifty one patients with human immuno-deficiency virus (HIV-1) infection who had been consecutively endoscoped for upper gastrointestinal symptoms were biopsied (stomach or duodenum, or both) and compared with 59 age and sex matched controls for the presence of Campylobacter pylori. In 28 (47%) of the control group but in only seven (14%) of the HIV seropositive patients were C pylori seen on histological examination (p less than 0.001, odds ratio 5.6, 95% confidence interval 2.2-14.5). Sixteen patients who were HIV antibody positive had other index diseases for the diagnosis of AIDS in the biopsy material and, when these were excluded, comparison with the control group still showed a significant difference; p less than 0.01, odds ratio 3.6, 95%, confidence interval 1.4-9.6. In this series, therefore, C pylori were far less common in HIV antibody positive patients than in controls. Among the HIV positive patients, a higher proportion of C pylori negative cases had AIDS but this trend was not significant. The findings of this study indicate that whatever abnormalities of cell mediated mucosal immunoregulation are caused by HIV infection, they do not seem to be important in the response to infection by C pylori.
Viral and toxoplasma gondii infections in children after liver transplantation.
Authors: Salt A , Sutehall G , Sargaison M , Woodward C , Barnes N D , Calne R Y , Wreghitt T G
Published: June 2007

The incidence and morbidity of viral and Toxoplasma gondii infections were studied in 40 children who underwent liver transplantation between December 1983 and February 1988. The incidence of primary and reactivated cytomegalovirus (CMV) infection was 19% and 47%, respectively; primary infection caused clinical disease in all five cases affected and was fatal in one. Primary Epstein-Barr virus (EBV) infection occurred in 10 (26%) recipients but caused only mild disease. No reactivated EBV infection was recorded and no lymphoproliferative disorders associated with EBV were found after a maximum of four years' follow up. Adenovirus infection occurred in seven (18%) patients; this was associated in one case with fatal pneumonia and fulminant hepatitis, but otherwise with only mild respiratory disease. Primary T gondii infection was detected in one patient who remained asymptomatic. Other viruses causing infection included herpes simplex, varicella zoster, and respiratory syncytial virus. Surveillance for these infections and the long term sequelae should be included in the follow up of all children who undergo transplantation.
Use of immunoelectron microscopy to show Ebola virus during the 1989 United States epizootic.
Authors: Geisbert T W , Jahrling P B
Published: June 2007

A filovirus, serologically related to Ebola virus, was detected by "post-embedment" immunoelectron microscopical examination of MA-104 cells. These had been infected by inoculation with serum samples obtained during the 1989 epizootic in cynomolgus monkeys (Macaca fascicularis), imported from the Philippines and maintained at Reston, Virginia, USA, a primate holding facility. The immunoelectron microscopy method, when used in conjunction with standard transmission electron microscopy (TEM) of infected cells, provided consistent results and was simple to perform in this epizootic. It is concluded that immunoelectron microscopy is potentially useful in the direct immunological diagnosis of Ebola and related filoviral infections (such as Marburg) in clinical samples obtained from those with acute infection.
Computer assisted selection and assessment of antibodies in the diagnosis of lymphomas.
Authors: Harkin P J , Kelly S A , Jack A S
Published: June 2007

With increasing numbers of reagents the problem of selecting appropriate antibodies to solve problems in the diagnosis of lymphoma is becoming more complex. One approach is to use a computer program to optimise the selection process. Such a program was devised, incorporating data from an extensive literature search. When presented with a differential diagnosis it selects the most appropriate antibody panel and when given the results evaluates the relative likelihood of each possible diagnosis. In a retrospective study 81% of the tests used had been non-discriminatory, but using the results of the remaining 19% of the tests, the computer was able to select the "correct" diagnosis with a high degree of certainty. The development and use of this system illustrated several problems in the application of computer assisted diagnostic techniques in histopathology. These problems include incomplete data and lack of understanding of the process of histopathological diagnosis.
Cellular expression of lymphocyte function associated antigens and the intercellular adhesion molecule-1 in normal tissue.
Authors: Smith M E , Thomas J A
Published: June 2007

A detailed immunohistological analysis of normal tissues for the distribution of lymphocyte function-associated antigens (LFA) and the intercellular adhesion molecule-1 (ICAM-1) showed several hitherto unrecognised patterns of LFA-3 and ICAM-1 expression. The widespread, but not ubiquitous, distribution of LFA-3 contrasted with the more restricted distribution of ICAM-1. Among epithelial cells, all tissues which were ICAM-1 positive were also LFA-3 positive with the single exception that thymic cortical epithelium, in contrast to previous reports, expressed only ICAM-1. It was striking that LFA-3 molecules were absent in some tissues which are considered to be sites of immunological privilege (such as brain and testis), suggesting an additional mechanism by which these microenvironments maintain immunological autonomy. Furthermore, the unexpected finding that LFA-3 is strongly expressed on intercalated discs of cardiac muscle may possibly be related to a non-immune function, or indicate a structurally similar epitope expressed by an unrelated molecule within this tissue.
Evaluation of a prototype for a reference platelet counter.
Authors: Lewis S M , Rowan R M , Kubota F
Published: June 2007

A semiautomated single channel aperture-impedance particle counter was developed as a prototype for a reference platelet count for assigning values to reference preparations used in automated blood cell counts. The instrument is equipped with sheath flow and an aperture orifice of 50 microns in diameter and 60 microns in length to eliminate non-axial flow and minimise coincidence errors. Use of fixed volume and red blood cell:platelet ratios obviate dilution errors. The counter was assessed in accordance with the International Committee for Standardization in Haematology protocol for evaluation of automated blood cell counters. The counter provided a high level of linearity and precision, accurate coincidence correction, controlled volume, stability and negligible carryover.
Erythroid colony growth from peripheral blood and bone marrow in polycythaemia.
Authors: Masters G S , Baines P , Jacobs A
Published: June 2007

Erythroid colony growth in the presence and absence of erythropoietin was compared in 23 patients with primary proliferative polycythaemia (PPP), nine with idiopathic erythrocytosis, 10 with secondary polycythaemia, 15 with pseudopolycythaemia and in 76 normal subjects. Erythroid colonies growing without erythropoietin stimulation (endogenous erythroid colonies) from peripheral blood (BFU-E) were found in 20 of 22 patients with PPP and in two of seven with idiopathic erythrocytosis. None was found in secondary polycythaemia, pseudopolycythaemia, or in normal subjects. Small numbers of endogenous colony forming units-erythroid (CFU-E) (though not BFU-E) were cultured from the bone marrow of three of 24 normal subjects, suggesting that peripheral blood cultures provide a more specific indicator of clonal erythropoiesis. Peripheral blood endogenous erythroid colony growth is an effective and convenient means of distinguishing patients with clonal erythrocytosis and may be of particular value when iron deficiency obscures the diagnosis of PPP on conventional criteria.
Lectin typing of Campylobacter isolates.
Authors: O'Sullivan N , Benjamin J , Skirrow M B
Published: June 2007

Isolates of Campylobacter jejuni, C coli, C fetus and C laridis were tested for agglutination reactions with a panel of five lectins: Arachis hypogaea, Bauhinia purpurea, Solanum tuberosum, Triticum vulgaris and Wisteria floribunda. Twenty three patterns of agglutination (lectin types) were recorded among 376 isolates. Patterns were consistent and reproducible. Only 4.5% of isolates were untypable because of autoagglutination. Some lectin types were found exclusively or predominantly in a species, but others were shared between species. Forty two per cent of C jejuni and 35% of C coli isolates belonged to lectin type 4. There was no apparent correlation between lectin type and serotype; different lectin types were found among strains of single Penner and Lior serotypes. Lectin typing is a simple and economical procedure suitable for use in non-specialist laboratories, either as an adjunct to serogrouping or, after further development, as a sole typing scheme.
Rheumatoid lymphadenopathy: a morphological and immunohistochemical study.
Authors: Kondratowicz G M , Symmons D P , Bacon P A , Mageed R A , Jones E L
Published: June 2007

Sixteen lymph nodes from 14 patients with rheumatoid arthritis were examined immunohistochemically and morphometrically and compared with 10 control nodes showing follicular hyperplasia from patients without rheumatoid disease. Frozen material was available from nine patients and all controls. The nodes from patients with rheumatoid arthritis seemed to share characteristic features. The most striking of these was follicular hyperplasia in which the germinal centres, in spite of being quite large, showed relatively sparse proliferative activity. The nodes often showed infiltration of germinal centres by CD8 positive T lymphocytes and contained fewer IL2R positive cells in the paracortex than controls. These and other features may have some correlation with disease activity. Lymphadenopathy in rheumatoid arthritis may not just be a manifestation of joint inflammation but an active component of this multisystem disease and may reflect a widespread immunological abnormality.
Oestrogen receptors in colorectal carcinoma.
Authors: Dawson P M , Shousha S , Blair S D , Carter G D , Jones J , Alaghband-Zadeh J , Theodorou N A
Published: June 2007

The oestrogen receptor content of colorectal adenocarcinoma was investigated using an established ligand binding biochemical assay and two more recently introduced techniques using specific monoclonal antibodies (Abbott ER-EIA and ER-ICA assay kits). Twenty nine tumours were investigated by the ligand binding assay. Only one (3.4%) tumour gave a weakly positive result (11 fmol/mg cytosol protein); the rest were all negative. Where sufficient tissue was available, the receptors were also determined by a quantitative immunoassay in 18 patients and an immunohistochemical method in 13 patients. The results were similarly all negative. It is concluded that most colorectal carcinomas, irrespective of sex, are oestrogen receptor negative, and it is thus unlikely that hormonal manipulation would have an influence on the course of the disease.
Radiation and other pathological changes in breast tissue after conservation treatment for carcinoma.
Authors: Girling A C , Hanby A M , Millis R R
Published: June 2007

Histological changes in breast tissue after radiotherapy were evaluated. Changes in tissue from 17 patients previously treated for breast cancer by surgery and radiotherapy were compared with those seen in a control group treated with surgery alone. Vascular and epithelial changes were seen only in tissue from patients who had received radiotherapy and, therefore, seemed to be relatively specific; stromal changes were seen in both groups and seemed, therefore, to be non-specific. Epithelial atypia is of particular importance as it may be severe and be confused with recurrent malignancy. The presence of other changes associated with radiotherapy, particularly those in vessels, should help to avoid such misdiagnosis.
Effect of haemolysate preparation on measurement of red cell folate by a radioisotopic method.
Authors: Gilois C R , Stone J , Lai A P , Wierzbicki A S
Published: June 2007

The effects of varying the method of preparation of haemolysates on the measurement of red cell folate concentration were investigated using the Simultrac kit. The concentration of ascorbic acid did not have any significant effect on the assayed concentrations of folate, but lower concentrations were obtained when the incubation time was increased. Folate was stable for 14 days in cells when they were stored at 4 degrees C and for seven days at -25 degrees C, but instability was increased by storage in ascorbic acid, by the use of stored (4 degrees C) ascorbic acid, and by preparing the haemolysates by freeze-thaw cycling. It is recommended that haemolysates should be diluted in fresh ascorbic acid, at a concentration of 10 g/l, incubated for 60 minutes in the dark and stored at -20 degrees/25 degrees C before being assayed.
Aplastic anaemia associated with organochlorine pesticide: case reports and review of evidence.
Authors: Rugman F P , Cosstick R
Published: June 2007

Three patients with aplastic anaemia had a history of substantial previous exposure to organochlorine pesticides. The temporal association between chemical exposure and the onset of first symptoms of anaemia was strongly supportive. Organochlorines have the property of lipid affinity and accumulation in adipose tissue. Objective evidence of clinically important concentrations of tissue pesticide residues may be a useful confirmation of previous exposure. In the patients studied the presence of Lindane (gamma hexachlorocyclohexane) was shown using gas chromatography/mass spectrometry with selective ion monitoring of fragments obtained from one heavily exposed patient, with concentrations about five times greater than a matched control. The presence of clinically important tissue concentrations of pentachlorophenol was also confirmed in a second patient exposed to this agent. The long term safety of organochlorine pesticides remains doubtful as they were introduced before adequate toxicological screening tests had been developed. The central registration of possible haematological adverse reactions, however, forms an important epidemiological method in the study of environmental chemical hazards and should be complied with whenever possible.
Mast cells in leprosy skin lesions.
Authors: Cree I A , Coghill G , Swanson Beck J
Published: June 2007

The variability of mast cell density within and between leprosy skin lesions was examined as a basis for future studies, and whether the number of mast cells in the lesion was determined by local or systemic factors was evaluated. The mast cell density in the granuloma, skin appendages, and intervening dermis was assessed by counting mast cells in glycol methacylate sections stained with Giemsa stain and relating these counts to area measurements obtained by planimetry. In biopsy specimens taken from the edge of established lesions the density of mast cells within the granulomata was considerably higher than that in the intervening dermis and was comparable with that found in the appendages. No major differences in mast cell density were found between unaffected skin and the centre or edge of individual lesions. Mast cell densities in biopsy specimens from the edge of different lesions on the same patient were also similar, suggesting that the mast cell density within the granulomata is independent of the site of the lesion and is determined systemically.
Ber-EP4: new monoclonal antibody which distinguishes epithelia from mesothelial.
Authors: Latza U , Niedobitek G , Schwarting R , Nekarda H , Stein H
Published: June 2007

A new monoclonal antibody, Ber-EP4, directed against a partially formol resistant epitope on the protein moiety of two 34 kilodalton and 39 kilodalton glycopolypeptides on human epithelial cells is described. Immunostaining of a wide range of normal and neoplastic human tissues and cell lines showed that all carcinomas and all non-neoplastic epithelial cells, except hepatocytes, parietal cells, and apical cell layers in squamous epithelia, homogeneously expressed Ber-EP4 antigen. As Ber-EP4 does not detect any normal or neoplastic non-epithelial cells, this antibody might prove valuable for the differentiation of the following (i) non-epithelial tumours from undifferentiated carcinomas; (ii) hepatocytes from bile duct cells in certain liver diseases; (iii) mesothelial cells from carcinoma cells in lung biopsy specimens; and (iv) reactive mesothelial cells from carcinoma cells in smears of serous effusions.
Immunohistological study of oestrogen receptors in breast carcinomas that are biochemically receptor negative.
Authors: Shousha S , Stamp T , James K R , Alaghband-Zadeh J
Published: June 2007

The reliability of an immunohistological method, applied to paraffin wax sections, was assessed for determination of oestrogen receptor content of biochemically oestrogen receptor negative breast carcinomata. Sixty consecutive tumours with oestrogen receptor concentrations of less than 10 fmol/mg cytosol protein, as estimated by dextran-coated charcoal biochemical assay, were examined. Paraffin wax sections were treated with DNAse before applying a peroxidase-anti-peroxidase method using ER-ICA monoclonal antibodies. Fifty one cases (85%) were negative, six (10%) weakly positive, and three (5%) were moderately positive. No strongly positive cases were seen. It is suggested that cases with weakly positive staining, especially when localised to a small area, should be regarded as negative. On the other hand, as the three moderately stained cases included two small tubular carcinomas and an invasive ductal carcinoma with high progesterone receptor concentrations, it is more likely that the biochemical assay in these cases represented false negative results due to sampling error or inclusion of fibrous or other non-neoplastic tissue in the assayed samples. It is concluded that the immunohistological method used here is fairly reliable and would be especially valuable for determination of oestrogen receptor content in small, mammographically detected tumours from which no tissue would be available for biochemical assay or frozen section examination.
Rate nephelometric determination of rheumatoid factor: comparison between Kallestad QM-300 and Beckman ICS-II (RF) methods.
Authors: Collins R J , Neil J C , Wilson R J
Published: June 2007

The Kallestad Corporation recently suggested that their new buffer system for the nephelometric detection of rheumatoid factor conferred advantages over existing systems. Two rate nephelometric procedures, the Kallestad QM-300 and the Beckman ICS-II (RF), were therefore compared. Sera (n = 157) were selected on the basis of a previous ICS-II value. The results on the QM-300 of sera with an initial rheumatoid factor value of less than 400 IU identified two groups. Group 1 (n = 109) showed a good correlation with the ICS-II method while group 2 (n = 13) was highly discordant with the QM-300, producing significantly higher values. The values of 35 sera with an initial rheumatoid factor of greater than 400 IU were likewise highly discordant, with the QM-300 producing significantly lower values. Dilution recovery experiments implied that the Beckman buffer was likely to be contributory. As the formulae of the buffers remain proprietary, the reasons for the differences are speculative. The findings could be taken to indicate that the Kallestad value is a more accurate indicator of the quantity of rheumatoid factor than the Beckman value.
Coagulase testing compared with commercial kits for routinely identifying Staphylococcus aureus.
Authors: Rossney A S , English L F , Keane C T
Published: June 2007

Five commercial Staphylococcus aureus identification kits--Staphaurex (Wellcome), Staphylase (Oxoid), Staphyslide (bioMèrieux), Biostaph (Medlabs) and Bacto Latex (Difco)--were evaluated for the routine identification of S aureus from primary plates in the routine microbiology laboratory. Comparison was made with two methods of tube coagulase testing and five slide methods for detecting clumping factor (slide coagulase testing). Performances were assessed for two groups of organisms, staphylococcal species alone and a combined staphylococcal and non-staphylococcal species group. The effects of growth on selective media and storage of isolates at room temperature and 4 degrees C were investigated. Selective media cannot be recommended, nor can storage of isolates before testing. Ranked according to efficiency value with the combined staphylococcal and non-staphylococcal species group, the kits and coagulase methods performed as follows (the figures in parentheses are the efficiency values for the staphylococcal group alone): tube coagulase reference method 100% (100%), tube coagulase SJH method 99% (99%), Staphaurex 94% (97%), Staphylase 93% (96%), slide coagulase method No 4 93% (94%), slide coagulase method No 5 93% (93%), Bacto Latex 92% (95%), Staphyslide 92% (95%), and Biostaph 87% (91%). It is concluded that a commercial S aureus identification kit should not replace tube coagulase testing for the routine identification of the organism from primary plates and that, even the kits with the best performances, have little advantage over a good slide coagulase test method.
Immunohistochemical studies of intrahepatic tumour necrosis factor alpha in chronic liver disease.
Authors: Yoshioka K , Kakumu S , Arao M , Tsutsumi Y , Inoue M , Wakita T , Ishikawa T , Mizokami M
Published: June 2007

To determine the intrahepatic production of tumour necrosis factor alpha (TNF alpha) in chronic liver disease three monoclonal antibodies were used against TNF alpha in immunohistochemical studies of liver tissue sections from patients with chronic liver disease. All three monoclonal antibodies stained infiltrating mononuclear cells. Monoclonal antibody II 7C2 also stained the cytoplasm or nucleus, or both, of a varied number of hepatocytes from nine patients with chronic hepatitis B virus infection, suggesting that the antigenic epitope related to hepatitis B core antigen (HBcAg) crossreacted with II7C2. The other two monoclonal antibodies, III2F3 and IV3E5, stained significantly larger numbers of mononuclear cells in cases of chronic active hepatitis B than in chronic persistent hepatitis B, or hepatitis B related liver cirrhosis. III2F3 stained significantly larger numbers of mononuclear cells in non-A, non-B chronic active hepatitis than in chronic persistent hepatitis B or hepatitis B related liver cirrhosis. These results indicate that TNF alpha is produced and secreted by infiltrating mononuclear cells in focal inflammatory areas of the liver, and suggest that TNF alpha may have a role in the inflammatory activity of chronic liver disease.
Increased expression of laminin/collagen receptor (VLA-1) on epithelium of inflamed human intestine.
Authors: MacDonald T T , Horton M A , Choy M Y , Richman P I
Published: June 2007

Immunohistochemical techniques were used to investigate the epithelial expression of VLA-1 in inflammatory bowel disease in six patients with Crohn's disease, in four patients with ulcerative colitis, and in one patient with indeterminate colitis, and compared with that in the small intestine and colons of 10 normal controls. In normal small bowel VLA-1 was expressed on crypt epithelial cells and only weakly or not at all on surface epithelium. VLA-1 was again expressed weakly in normal colon, except in one case, a 1 year old child with diarrhoea but no histological abnormalities. In small and large intestine affected with Crohn's disease, ulcerative colitis, or indeterminate colitis, there was increased expression of VLA-1 on the basolateral aspects of crypt cells and de novo expression on surface epithelium. It is suggested that this is an adaptive response to prevent epithelial cell loss as a result of inflammation in the underlying lamina propria.
Evaluation of semiautomated procedure for lymphocyte subset analysis.
Authors: Bryson G J , Keegan R P , Quinn C P , Collins R J
Published: June 2007

The findings of recent surveys indicate a need for standardisation in lymphocyte subset analysis by flow cytometry. Major areas of concern are the methods used for labelling subsets and the choice of appropriate monoclonal antibodies. A standard dual colour manual whole blood lysis technique for flow cytometry was compared with the recently available Coulter Q-Prep EPICS technique. Overall, there was no significant difference (Student's t test) between the use of anticoagulated blood treated with heparin or EDTA. When normal subjects were examined there was a decrease in the absolute number of CD3+ and Leu-7+/CD8- cells and an increase in CD19+ and CD20+ cells. When human immunodeficiency virus (HIV) antibody positive subjects were examined there was a significant decrease in the absolute number of CD2+, CD3+, CD4+ and Leu 7+/CD8- cells and an absolute increase in CD19+ and CD20+ cells. CD8+ cells were decreased only with the Cyto-Stat reagents. Occasionally, the Q-Prep did not lyse the red cells efficiently. While the Q-Prep EPICS system has the potential to standardise and automate the labelling procedures used in lymphocyte subset analysis, further refinement, such as the choice of monoclonal antibodies or alternative preparative reagents, may be required to resolve the cause of the discordant findings between the two approaches.
Measurement of glomerular filtration rate in homozygous sickle cell disease: a comparison of 51Cr-EDTA clearance, creatinine clearance, serum creatinine and beta 2 microglobulin.
Authors: Aparicio S A , Mojiminiyi S , Kay J D , Shepstone B J , de Ceulaer K , Serjeant G R
Published: June 2007

Glomerular filtration rates (GFR) were measured with 51Cr-EDTA in 38 patients (aged 40-75 years) with homozygous sickle cell disease and compared with serum beta 2 microglobulin concentrations in 38 patients and with creatinine clearance in 21 patients. GFR estimated with 51Cr-EDTA was closely correlated with single serum creatinine measurements and the inverse of serum beta 2 microglobulin. Creatinine clearance was also found to be correlated, but values were, on average, 32% below those obtained by the 51Cr-EDTA method, and this difference was significant. It is concluded that measurements of beta 2 microglobulin, single serum creatinine, and creatinine clearance are valuable indicators of GFR in homozygous sickle cell disease. Measurement of beta 2 microglobulin was a useful and reliable method of estimating GFR from single plasma measurements and is therefore a useful means of screening the population.
Detection of a mucin marker for the adenoma-carcinoma sequence inhuman colonic mucosa by monoclonal antibody AM-3.
Authors: Hanski C , Bornhoeft G , Topf N , Hermann U , Stein H , Riecken E O
Published: June 2007

The monoclonal antibody AM-3 was raised against mucins extracted from human colorectal carcinomas. It reacted strongly with sections of paraffin wax embedded colorectal carcinoma. In colonic adenoma tissue the percentage of cells expressing the epitope detected by AM-3 correlated with the degree of dysplasia. In contrast to immunohistochemical staining, which did not show the presence of the antigen in histologically normal mucosa, the more sensitive enzyme linked immunosorbent assay (ELISA) and immunoblot assays showed that it was weakly expressed in this tissue. AM-3 reacted with variable frequency with several normal and malignant human tissues, indicating that the detected epitope is not restricted to colonic tissue. In colonic carcinomas it is present on a sialomucin of apparent relative molecular mass of more than 440,000. These data suggest that the antigen detectable with AM-3 may be useful in the assessment of premalignant changes in colonic adenomas.
Expression of epidermal growth factor receptor in breast carcinoma.
Authors: Lewis S , Locker A , Todd J H , Bell J A , Nicholson R , Elston C W , Blamey R W , Ellis I O
Published: June 2007

A series of 90 patients with primary operable breast cancer and with up to 36 months of follow up were investigated for expression of epidermal growth factor receptor (EGFR) in their tumours by immunocytochemical staining with the monoclonal antibody EGFR 1. Tumour samples were snap frozen in liquid nitrogen immediately after resection and subsequently stained using a standard indirect immunocytochemical method. Tumour staining was assessed by two observers and scored on a four point scale (0-3). Thirteen (14%) tumours showed positive immunoreactivity. A strong correlation between distinct EGFR expression and short disease free interval was observed. Significant correlations were also shown with oestrogen and progesterone receptor expression and tumour nuclear size. No significant association was found with tumour size, lymph node stage, and histological grade. The association with disease free interval remained significant in multivariate analysis.
In situ hybridisation in herpetic lesions using a biotinylated DNA probe.
Authors: Dictor M , Renfjärd E , Brun A
Published: June 2007

In situ hybridisation was performed with a biotinylated DNA probe for herpes simplex virus (HSV) using high temperature denaturation on formalin fixed, paraffin wax sections of lung, brain, ganglion and keratinising and non-keratinising squamous epithelia. Eosinophilic viral nuclear inclusions or characteristically moulded multiple nuclei with altered chromatin, which were present in two cases of HSV encephalitis and one case of viral pneumonitis, all showed complete hybridisation visualised by an alkaline phosphatase/nitroblue tetrazolium detector system. HSV encephalitis and trigeminal ganglionitis, which were confirmed serologically or clinicopathologically but lacked nuclear changes, also gave positive dense nuclear signal in neurons, glias and satellite cells. No staining was present in the ganglion cells in trigeminal zoster, the glia in progressive multifocal leucoencephalopathy, or in a variety of cells in a lung coinfected with cytomegalovirus. In 10 herpetic blisters of squamous epithelia, infected cells hybridised strongly, while morphologically similar herpes zoster lesions remained negative. In neural tissues non-hybridisation staining was most obtrusive in corpora amylacea and seemed to reflect nonspecific probe adherence. In squamous epithelium, major non-hybridisation staining was caused by probe and antibody possibly adhering to intracellular keratin. The HSV probe permits specific detection of virus in the absence of characteristic nuclear changes and allows varicella zoster virus to be differentiated from HSV, provided that the aforementioned problems with non-hybridisation staining are borne in mind.
Detection of messenger RNA using a digoxigenin end labelled oligodeoxynucleotide probe.
Authors: Farquharson M , Harvie R , McNicol A M
Published: June 2007

A synthetic oligodeoxynucleotide sequence complementary to the mRNA for the adrenocorticotrophin (ACTH) precursor pro-opiomelanocortin (POMC) was end labelled using digoxigenin. The probe was used to detect POMC mRNA both on nitrocellulose filters and by non-isotopic in situ hybridisation (NISH) in tissue sections. Digoxigenin was identified using anti-digoxigenin alkaline phosphatase. The model system examined was the rat pituitary gland. Removal of both adrenal glands and dexamethasone administration were used to change the concentrations of POMC mRNA in the rat anterior lobe. The labelled probe reacted with a single band of appropriate molecular weight in Northern blot analysis. The distribution of signal in tissue sections and the changes induced by experimental manipulation were as predicted. The results indicate that this method of NISH will prove useful in the detection of specific messenger RNAs in tissue sections of buffered, formalin fixed, paraffin wax embedded material.
Factor VIII inhibitor assay using skin puncture blood samples.
Authors: Hinchliffe R F , Bellamy G J , Lilleyman J S
Published: June 2007

A method was developed for the measurement of factor VIII inhibitors by the Bethesda technique using small volumes of skin puncture blood. Human and porcine inhibitor concentrations on paired venous and skin puncture samples measured on 10 occasions were highly correlated, with no significant difference between them. Assays on four inhibitor negative haemophiliac patients were also negative by the skin puncture method. Sample dilution at the collection stage results in loss of sensitivity to inhibitor concentrations below 1 unit/ml, but the technique will be of value in monitoring higher concentrations when venous access is difficult and must be saved for parenteral treatment.
Epithelial markers in pancreatic carcinoma: immunoperoxidase localisation of DD9, CEA, EMA and CAM 5.2.
Authors: Heyderman E , Larkin S E , O'Donnell P J , Haines A M , Warren P J , Northeast A , Grant A G
Published: June 2007

Paraffin wax embedded, formalin fixed sections of 22 adenocarcinomas of the exocrine pancreas were stained with four mouse monoclonal antibodies: DD9-E7, an antibody raised against a human pancreatic tumour xenograft; carcino-embryonic antigen (CEA); epithelial membrane antigen (EMA); and cytokeratin (CAM 5.2). An indirect immunoperoxidase technique without enzyme pre-digestion and an affinity-purified sheep anti-mouse peroxidase conjugate were used. All of the tumours were positive for DD9-E7, EMA, and CAM 5.2. Twenty out of 22 were focally positive for CEA and the staining was often weak. As all of these adenocarcinomas were DD9-E7 positive, absence of staining for DD9-E7 in a tumour makes the diagnosis of adenocarcinoma of the exocrine pancreas very unlikely, and this is of value in distinction from endocrine carcinomas with a marked acinar pattern. The weak CEA staining distinguished pancreatic carcinomas from colorectal tumours. Because the distribution of staining for EMA and CAM 5.2 was no different from that previously seen in adenocarcinomas from other sites, these markers are likely to be of limited value in the differential diagnosis of abdominal adenocarcinomas of uncertain origin.
Spontaneous mutation rate in retinoblastoma.
Authors: Morris J A
Published: June 2007

A novel approach was used to estimate the in vivo mutation rate of the retinoblastoma gene. A mathematical formula can be used to calculate the probability of neoplasia induced by one or more mutations in a population of dividing cells. This formula can then be applied to epidemiological data on hereditary and sporadic retinoblastoma. The analysis yields an estimate of the in vivo mutation rate of 8 x 10(-8)/gene/cell division (range 5.5 x 10(-8) to 1.3 x 10(-7]. The estimated non-replication association in vivo mutation rate is 2.4 x 10(-8)/gene/year (range 0 to 6.8 x 10(-8]. The formula is an improvement on previous attempts to produce a model of the process of mutation during cell generation. It can be applied to neoplastic disease in both children and adults.
Tissue extraction of DNA and RNA and analysis by the polymerase chain reaction.
Authors: Jackson D P , Lewis F A , Taylor G R , Boylston A W , Quirke P
Published: June 2007

Several DNA extraction techniques were quantitatively and qualitatively compared using both fresh and paraffin wax embedded tissue and their suitability investigated for providing DNA and RNA for the polymerase chain reaction (PCR). A one hour incubation with proteinase K was the most efficient DNA extraction procedure for fresh tissue. For paraffin wax embedded tissue a five day incubation with proteinase K was required to produce good yields of DNA. Incubation with sodium dodecyl sulphate produced very poor yields, while boiling produced 20% as much DNA as long enzyme digestion. DNA extracted by these methods was suitable for the PCR amplification of a single copy gene. Proteinase K digestion also produced considerable amounts of RNA which has previously been shown to be suitable for PCR analysis. A delay before fixation had no effect on the amount of DNA obtained while fixation in Carnoy's reagent results in a much better preservation of DNA than formalin fixation, allowing greater yields to be extracted.
Rapid assessment of haemophilia A carrier state by non-invasive techniques using the polymerase chain reaction.
Authors: Ball J , Warnock L J , Preston F E
Published: June 2007

The polymerase chain reaction (PCR) was used to amplify two polymorphic regions in the factor VIII gene. In vitro synthesis of DNA was achieved using samples obtained from buccal cells, urine, and hair follicles in addition to purified genomic and crude DNA samples prepared from whole blood. Female members of two kindreds affected with haemophilia A were assessed for carrier state using direct restriction fragment length polymorphism analysis of amplified gene products in the Bc1I and XbaI regions. It is concluded that this is a non-invasive, rapid, and inexpensive technique for carrier detection.
Comparison of an immunochemical assay for plasma fibrinogen and a turbidimetric thrombin clotting technique to discriminate hyperlipidaemic patients from healthy controls.
Authors: Knapp M L , Feher M D , Carey H , Mayne P D
Published: June 2007

Plasma samples from patients attending a lipid clinic (n = 14) and healthy control subjects (n = 21) were assayed for fibrinogen using an immunochemical method (radial immunodiffusion) and a turbidimetric assay based on the thrombin clotting technique. The patients had significantly higher plasma fibrinogen concentrations than controls by both methods, but there was significant overlap between the two groups when fibrinogen was assayed by the thrombin clotting technique; there was almost complete separation of the two groups using the immunochemical assay. This difference in overlap could not be attributed to the presence or absence of fibrinogen degradation products. These findings may have important implications for the choice of method for determining plasma fibrinogen when assays are used for the assessment of cardiovascular risk. It is recommended that plasma fibrinogen should be assayed by both an immuno-chemical and a thrombin clotting method.
Enzyme linked immunosorbent assay for detecting adenoviruses in stool specimens: comparison with electron microscopy and isolation.
Authors: Martin A L , Kudesia G
Published: June 2007

A commercial enzyme linked immunosorbent assay (ELISA) for detection of adenoviruses in stool samples was compared with the use of electron microscopy and isolation in Graham 293 cells. Although specific, the ELISA was less sensitive than both electron microscopy and isolation. The ELISA had an overall sensitivity of 78% and a specificity of 100%. The sensitivity was related to the amount of virus particles present in the stool sample, increasing to 90% with about 10(7) viral particles/ml of stool. The ELISA was easy to perform, requiring no instrumentation, and is a useful first line test for detection of adenoviruses in stool samples, especially in laboratories without access to an electron microscope. Wider use of ELISAs should help in evaluating the role of adenoviruses in viral gastroenteritis.
Technique for identifying cutting artefacts in sections of undecalcified bone biopsy specimens.
Authors: Wood D J , Mawhinney W H , Malcolm A J , Stevens J
Published: June 2007

Areas of fragmentation found in trabecular and cortical bone of iliac crest biopsy specimens have been described as bone quality defects and were thought to be the major factor responsible for femoral neck fractures. These appearances have also been regarded as cutting artefacts and to resolve this difference of opinion consecutive sections cut at right angles to each other in biopsy specimens from 15 patients with femoral neck fractures were compared. Sections were assessed by four independent observers; agreement by at least three was required before an area was accepted as a bone quality defect. In all, 270 were identified. Of the 161 found in sections cut parallel to the cortices, there were only 20 (12%) in coincident areas in consecutive sections. This study shows conclusively that areas of fragmentation previously described as bone quality defects are not artefacts which may be created or excluded depending on the plane of section.
Expression of CD15 antigen in urinary bladder transitional cell carcinoma.
Authors: Parham D M , Morton K , Coghill G , Robertson A J , Kerr M A
Published: June 2007

The biopsy specimens of 91 patients between the ages of 38 and 94 with transitional cell carcinoma of the bladder were retrospectively reviewed to determine if the expression of CD15 antigen detected by a monoclonal antibody MC2 was correlated with prognosis. Expression was variable, ranging from strong expression of the antigen by only the superficial cells in well differentiated papillary lesions to weak expression by most cells in solid or invasive tumours. In the invasive component there was a correlation between MC2 expression and tumour type, suggesting that the cell surface carbohydrate detected by MC2 may have a role in cell adhesion. There was no correlation between staining and survival. It is concluded that tumour type, grade, and stage remain the best prognostic indicators of urothelial tumours.
Antibody response of fibrocystic patients to homologous 0-typable and 0-defective isolates of Pseudomonas aeruginosa.
Authors: MacDougall J , Hodson M E , Pitt T L
Published: June 2007

Antibody titres to Pseudomonas aeruginosa of sera from 60 adult fibrocystic patients were determined in an enzyme linked immunosorbent assay (ELISA) with whole cells of homologous isolates which had been classified according to 0-antigen state by their reactivity with 0-typing antisera. Patients who were continuously colonised with Ps aeruginosa gave the highest titres: range 1500-64000 (mean 11000) and 500-48000 (mean 9000) with homologous 0-typable and 0-defective isolates, respectively. Lower titres to both varieties of isolates were obtained with recently colonised patients, and non-colonised patients gave titres with reference laboratory strains marginally above those of healthy controls. Serum titres of patients with sequential isolates were strain dependent and did not correlate with the 0-antigen state of the strain. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblot analysis of these sera and strains showed antibody binding primarily to high molecular weight 0-repeating units of lipopolysaccharide. It is concluded that the 0-antigen of the strain of Ps aeruginosa used in the ELISA test does not influence the titre obtained with fibrocystic sera, and it is recommended that serum titres should be assessed with a panel of homologous isolates from patients.
Evaluation of two rapid methods for identifying and biotyping Haemophilus influenzae.
Authors: Murphy P G , Craig I , Lafong A C , Smyth E T
Published: June 2007

Several rapid method kits (one to four hours) have become available for the identification of Haemophilus and related genera. Two kits (the "Rapid NH" system and the "RIM Haemophilus" system), which include the identification and biotyping of H influenzae, were investigated for the rapid identification and biotyping of 193 isolates of H influenzae and the results compared with those obtained by more standard overnight methods. The kits were convenient to use and gave reliable and rapid speciation of all isolates. Both test systems were unreliable for biotyping: 42 isolates were wrongly biotyped by the RIM kit and 40 isolates wrongly biotyped by the rapid NH kit. It is concluded that the test kits may be useful for the rapid identification of H influenzae but that they are not reliable for the biotyping of this species.
Application of immunoblotting to detect soluble Pneumocystis carinii antigen(s) in bronchoalveolar lavage of patients with Pneumocystis pneumonia and AIDS.
Authors: Sethi K K
Published: June 2007

The technique of immunoblotting for detecting soluble Pneumocystis carinii antigen(s) in bronchoalveolar lavage fluid specimens from patients with AIDS and Pneumocystis pneumonia was evaluated. A soluble 67 kilodalton polypeptide that was immunoreactive with an anti-P carinii monoclonal antibody (2G2) was found in the supernatants of 26 lavage samples from patients with pneumocystosis. Intact organisms in lavage sediments were detected by methenamine silver or immunofluorescence staining procedures. The diagnostic use of this technique was shown in four cases in which lavage sediments proved negative for intact Pneumocystis carinii organisms on first examination; 2G2 reactive soluble antigen, however, was identified in the immunoblots of the supernatants from the same samples. It is concluded that immunoblotting of bronchoalveolar lavage specimens using 2G2 monoclonal antibody as a detection probe may be a useful adjunct to the morphological demonstration of organisms by special staining procedures.