bluuebunny/crossref_metadata_2025 huggingface dataset:
Expression of colonic antigens by goblet and columnar epithelial cells in ileal pouch mucosa: their association with inflammatory change and faecal stasis.
Authors: Campbell A P , Merrett M N , Kettlewell M , Mortensen N J , Jewell D P
Published: June 2007

AIMS--To investigate colonic metaplasia of goblet and columnar epithelial cells in ileal pouch mucosa; to correlate this with the degree of morphological and inflammatory change; and to assess whether such changes are related to the presence of faecal stasis. METHODS--Biopsy specimens of ileal pouch mucosa were taken from 31 patients (30 with ulcerative colitis, one with familial adenomatous polyposis) either before (eight patients) or after (23 patients) ileostomy closure. A simple morphological technique was used to assess changes in villous height. Inflammatory change was estimated using an established scoring system for pouchitis, and acquisition of colonic antigens was determined by immunohistochemistry using three monoclonal antibodies which recognise components of the two major epithelial cell types in the colorectum. The degree of staining with the monoclonal antibodies was graded and the grades correlated with an index of villous atrophy and with the inflammatory scores. RESULTS--Five of eight (63%) pre-closure and 15 of 23 (65%) post-closure biopsy specimens showed increased staining with an antibody against components of columnar epithelial cells. One of eight (12%) pre-closure and 15 of 23 (65%) post-closure biopsy specimens stained with an antibody for colonic mucin. Although both types of staining showed a positive correlation with the pouchitis score, they also occurred in the absence of inflammation. CONCLUSIONS--Both goblet and columnar cells acquire colonic characteristics which are incomplete, but may represent a true adaptive response as they can develop in the absence of inflammation. As the change in goblet cells occurs after ileostomy closure, faecal stasis is likely to be a major contributory factor. Changes in columnar cells may occur before ileostomy closure in the absence of faecal stasis.
Xanthoderma: an unusual presentation of hypothyroidism.
Authors: al-Jubouri M A , Coombes E J , Young R M , McLaughlin N P
Published: June 2007

A young woman presented with progressive yellowing of her skin over a period of six months. Liver function tests were requested by her general practitioner and the results prompted the Chemical Pathology Department to instigate further tests to reach the final diagnosis. Hypercarotenaemia had caused her yellow skin, and various other biochemical abnormalities pointed towards primary hypothyroidism as an underlying cause. Thyroxine replacement treatment successfully corrected all the biochemical abnormalities including hypercarotenaemia. As far as is known, yellow skin as a sole presenting feature of hypothyroidism is extremely rare.
Paget's disease of the breast in a man without underlying breast carcinoma.
Authors: O'Sullivan S T , McGreal G T , Lyons A , Burke L , Geoghegan J G , Brady M P
Published: June 2007

A case of histologically confirmed Paget's disease of the breast in a 72 year old man, without underlying breast carcinoma, is reported. This report raises questions about the pathogenesis of this condition and suggests that Paget's disease is an independent, intraepidermal carcinoma rather than a direct extension of intraductal carcinoma of the breast to the nipple and areola.
Use of a monoclonal antibody and a DNA probe for diagnosing acute toxoplasmosis in ambiguous cases.
Authors: Garberi J C , Angel S O , Paulin P , Pszenny V , Romano L
Published: June 2007

An unusual case of cerebral toxoplasmosis in an HIV negative 11 year old child is reported. Central nervous system disease was assessed immunohistochemically in a brain biopsy specimen with TgP8--a specific monoclonal antibody against Toxoplasma gondii antigens--thus confirming IgG and IgM serology, technetium scan findings, and clinical data. In addition, an active parasitaemia was confirmed by DNA in situ hybridisation assay in white cells using an ABGTg4 probe. The child recovered after specific T gondii treatment. Follow up six months later showed that he was immunodeficient.
Quantification of HIV by PCR in monocytes and lymphocytes in patients receiving antiviral treatment and low dose recombinant human granulocyte macrophage colony stimulating factor.
Authors: Davison F D , Kaczmarski R S , Pozniak A , Mufti G J , Sutherland S
Published: June 2007

Neutropenia induced by antiviral treatment, in particular AZT, can be improved with recombinant human granulocyte macrophage colony stimulating factor (RHGMCSF) in HIV positive patients. However, there has been concern that this may increase the HIV load in the mononuclear cells of such patients. Five patients receiving AZT plus low dose RHGMCSF are reported. There was no consistent change in the levels of HIV DNA in the monocytes and lymphocytes. Additionally, all patients had stable disease with no opportunistic infection during the study period. It is concluded, therefore, that low dose RHGMCSF does not significantly change the viral DNA load in patients receiving AZT.
In situ hybridisation for the identification of Helicobacter pylori in paraffin wax embedded tissue.
Authors: Bashir M S , Lewis F A , Quirke P , Lee A , Dixon M F
Published: June 2007

A method for identifying Helicobacter pylori using a non-isotopic in situ hybridisation technique is described. A probe generated by polymerase chain reaction (PCR) with primers directed against parts of the Helicobacter pylori 16SrRNA sequence was used. Paraffin wax embedded gastric biopsy specimens from patients with and without gastritis were hybridised with the probe, and the method was shown to be sensitive and specific for H pylori.
Tissue diagnosis of intestinal microsporidiosis using a fluorescent stain with Uvitex 2B.
Authors: Franzen C , Müller A , Salzberger B , Fätkenheuer G , Eidt S , Mahrle G , Diehl V , Schrappe M
Published: June 2007

AIMS--To detect intestinal microsporidiosis in paraffin wax embedded biopsy specimens using a fluorescence technique incorporating optical brighteners. METHODS--Eight HIV infected patients with confirmed intestinal microsporidiosis (six with Enterocytozoon bieneusi, one with Encephalitozoon intestinalis and one with Encephalitozoon cuniculi infection) and 10 without infection were studied. Tissue sections of paraffin wax embedded duodenal biopsy specimens were stained with 1% Uvitex 2B, coded and analysed independently by two investigators. RESULTS--In all eight cases with confirmed intestinal microsporidian infection, spores could be detected easily in tissue sections using the fluorescence technique. Spores or other elements consistent with microsporidiosis were not found in the 10 patients without infection. CONCLUSION--Staining of tissue sections from paraffin wax embedded intestinal biopsy specimens with stains incorporating Uvitex 2B is a rapid and easy technique for the diagnosis of intestinal microsporidiosis.
The polymerase chain reaction in the demonstration of monoclonality in T cell lymphomas.
Authors: Diss T C , Watts M , Pan L X , Burke M , Linch D , Isaacson P G
Published: June 2007

AIMS--To evaluate polymerase chain reaction (PCR) amplification of T cell receptor (TCR) beta and gamma chain genes as a means of demonstrating monoclonality in T cell lymphomas using histological samples; to compare the performance of PCR with Southern blot analysis. METHODS--TCR-beta, TCR-gamma and immunoglobulin heavy chain (IGH) genes were analysed using PCR in 55 cases of T cell lymphoma (28 frozen tissue and 27 paraffin wax embedded samples), diagnosed using morphological and immunohistochemical criteria. The 28 frozen samples were subjected to Southern blot analysis using TCR-beta, TCR-gamma and IGH gene probes. Twenty five B cell lymphomas and 21 non-neoplastic lymphoid tissue samples were used as controls. RESULTS--Using TCR-beta PCR, monoclonality was detected in 24 (44%) of 55 T cell lymphomas compared with 43 (78%) of 55 using TCR-gamma PCR and in 82% with both techniques. Five (9%) of 55 T cell lymphomas were IGH PCR positive. None of the non-neoplastic lymphoid control samples were PCR positive. All B cell lymphomas showed a polyclonal pattern with TCR-beta PCR while a single B cell lymphoma was positive using TCR-gamma primers. With TCR-beta PCR, a monoclonal result was seen in 12 (43%) of 28 frozen samples of T cell lymphoma, compared with 23 (82%) of 28 using Southern blot analysis. With TCR-gamma PCR, 19 (68%) of 28 frozen tissue samples were positive, compared with 26 (93%) of 28 using Southern blot analysis. A single case showed IGH rearrangement by Southern blot analysis. CONCLUSION--TCR-gamma PCR should be the method of choice for analysis of clonality in paraffin wax embedded sections of lymphoproliferative lesions, as TCR-beta PCR has a high false negative rate. Southern blot analysis remains the most successful technique when sufficient fresh tissue samples and resources are available.
Elevated serum alpha-fetoprotein in a patient with undifferentiated carcinoma of the gall bladder.
Authors: Ng W K , Ng W F
Published: June 2007

An uncommon case of undifferentiated carcinoma of the gall bladder in a 65 year old Chinese man, who presented with an increased serum alpha-fetoprotein concentration, is reported. Histologically, the tumour had a primitive appearance and was composed of a pavement-like array of poorly differentiated columnar/polygonal cells. Alpha-fetoprotein was demonstrated in some of the tumour cells using an immunoperoxidase technique. Alpha-fetoprotein secretion in this instance may have occurred because the gall bladder and the liver are of similar embryological origin. Alpha-fetoprotein may also be related to the resurgent expression of oncofetal antigens. This tumour may represent another rare cause of increased serum alpha-fetoprotein concentrations.
Avidity of specific IgG antibodies elicited by immunisation against Haemophilus influenzae type b.
Authors: Agbarakwe A E , Griffiths H , Begg N , Chapel H M
Published: June 2007

AIM--To investigate the avidity of specific IgG polyribosyl ribitol phosphate (PRP) antibodies induced by three Haemophilus influenzae type b (Hib) conjugate vaccines: PRP-meningococcal outer membrane protein complex (PRP-OMP), PRP-non-toxic mutant diphtheria toxin, CRM197 (HbOC) and PRP-tetanus toxoid (PRP-T). METHODS--One hundred and ten infants were immunised with one of the vaccines at two, three and four months of age. Blood samples were taken after each vaccination and serum stored at -20 degrees C. Serum samples collected after the third course (that is, at five months of age) were submitted to antibody avidity assessment, using a urea enzyme linked immunosorbent assay (ELISA). RESULTS--All three conjugate vaccines elicited IgG PRP antibodies of high median avidity. The resultant antibody populations were heterogeneous with regard to avidity, which in turn was independent of PRP antibody concentration. CONCLUSIONS--With the recent findings of a correlation between bactericidal killing and affinity, our data highlight the need for a protective level to be expressed qualitatively as well as quantitatively.
Use of the Pastorex aspergillus antigen latex agglutination test for the diagnosis of invasive aspergillosis.
Authors: Hopwood V , Johnson E M , Cornish J M , Foot A B , Evans E G , Warnock D W
Published: June 2007

AIMS--To evaluate the Pastorex aspergillus antigen latex agglutination test for the diagnosis of invasive aspergillosis in patients undergoing liver or bone marrow transplantation. METHODS--Serum samples were taken at least twice weekly post-transplant and tested for Aspergillus antigen. Latex agglutination test results were compared with microbiological examination of respiratory, urine and bile specimens. Serum samples from liver transplant patients were also tested for antibodies to Aspergillus fumigatus by counter immunoelectrophoresis. RESULTS--Eight of the 91 patients studied developed invasive aspergillosis. Positive latex agglutination tests were obtained in eight of 187 (4.3%) serum samples from four of these eight patients. The other four patients with invasive aspergillosis gave consistently negative latex agglutination tests. A positive latex agglutination test was the first indication of invasive aspergillosis in two patients; these patients were already on amphotericin B. Positive latex agglutination tests were the only evidence of invasive aspergillosis in one patient who subsequently died of the infection. False positive latex agglutination tests were obtained in five of 83 (6%) patients with no evidence of invasive aspergillosis and misleading positive cultures seen in nine of 83 (10.8%). No antibodies were detected in three of four liver transplant patients with invasive aspergillosis. Conversely, antibodies were detected in 63 of 262 (24%) serum samples from 43 liver transplant patients with no evidence of invasive aspergillosis; one of these patients had an antibody titre of 1:2 on four separate occasions. CONCLUSIONS--The Pastorex aspergillus antigen latex agglutination test, when used alone, lacks sensitivity and specificity for the early diagnosis of invasive aspergillosis. A diagnosis was made in all patients with invasive aspergillosis when both culture and antigen tests were performed although using these criteria a false positive diagnosis would have been made in 13 of 83 (15.6%) patients. Microbiological and serial serological investigations for antigen should both be performed and the results considered in conjunction with radiological and clinical evidence.
Molecular markers for diagnostic cytology of neoplasms in the head region of the pancreas: mutation of K-ras and overexpression of the p53 protein product.
Authors: van Es J M , Polak M M , van den Berg F M , Ramsoekh T B , Craanen M E , Hruban R H , Offerhaus G J
Published: June 2007

AIMS--To determine the potential efficiency of molecular markers specific for neoplastic change--mutations of the K-ras oncogene and the p53 tumour suppressor gene--in diagnosing pancreatic carcinoma. METHODS--Archival cytology samples obtained from 17 patients with established pancreatic carcinoma were assayed for alterations in K-ras and p53. To detect changes in p53 expression, immunocytochemistry with polyclonal antibody CM1 was performed on the archival cytology slides after destaining. Mutations in K-ras codon 12 were then analysed on the scrapings of the same slides using mutant enriched polymerase chain reaction (PCR) amplification and restriction fragment length polymorphism analysis with allele specific oligonucleotide hybridisation for confirmation and characterisation. RESULTS--False negative results were recorded for five of the cytology slides when compared with p53 immunostaining of the surgical resection specimen. These five cases had been stained previously with Giemsa which interacts adversely with the immunostaining in contrast to the Papanicolaou procedure. The K-ras codon 12 mutations followed the well established distribution frequency and spectrum for pancreatic cancer and corresponded with the findings in the resection specimens in all cases. Two scrapings yielded insufficient DNA for PCR. Importantly, for two cases with an inconclusive cytology diagnosis on routine light microscopy, the diagnosis was confirmed by one of the molecular markers. The application of the molecular markers increased the diagnostic accuracy of cytology in this small study from 76 to 89%. CONCLUSIONS--The study indicates that assessment of alterations in the K-ras and p53 genes may be a valuable adjunct to diagnostic cytopathology of the head region of the pancreas, although there are some difficulties which will have to be overcome.
Histomorphometric classification of postmenopausal osteoporosis: implications for the management of osteoporosis.
Authors: Rehman M T , Hoyland J A , Denton J , Freemont A J
Published: June 2007

AIMS--To define and group static and dynamic iliac crest histomorphometric parameters in women with established osteoporosis. METHODS--Iliac crest biopsy specimens from 146 white women were sectioned undecalcified and examined using image analysis. RESULTS--Five distinct groups were defined on the basis of histomorphometric changes in cell function: group 1, decreased osteoblastic and osteoclastic activity; group 2, decreased osteoblastic and increased osteoclastic activity; group 3, increased osteoblastic and osteoclastic activity; group 4, no bone surface cell activity; and group 5, apparently normal osteoblastic and osteoclastic activity. CONCLUSIONS--Five distinct subgroups of patients with postmenopausal osteoporosis can be defined based on changes in bone cell function. Defining cellular dysfunction in this way may be important for tailoring treatment regimens to the needs of individual patients.
Audit of tumour histopathology reviewed by a regional oncology centre.
Authors: Prescott R J , Wells S , Bisset D L , Banerjee S S , Harris M
Published: June 2007

AIMS--To analyse the diagnostic differences in reporting tumour histopathology between a district general hospital and a regional oncology centre. METHODS--Tumour histopathology reports (n = 227) extracted from Bolton General Hospital files between 1988 and 1992 were compared with the corresponding Christie Hospital (oncology centre) reports, the same material having been seen at both hospitals. RESULTS--Diagnostic agreement existed in 77% of all cases. The incidence of major discrepancies was 8.37%. Of the diagnoses, 19 (36%) cases involved major discrepancies and 34 (64%) cases minor discrepancies. Most discrepancies occurred in the lymphoma group and involved subclassification of Hodgkin's and non-Hodgkin's lymphoma. Ki1 anaplastic large cell lymphoma and T cell rich B cell lymphoma were problematic diagnoses. The correct grading of follicle centre cell lymphomas using the Kiel classification was another problem area. In 19 cases certain aspects of immunohistochemistry produced discrepancies. In one case an incorrect diagnosis was made at the oncology centre and in another both centres gave an incorrect diagnosis. CONCLUSIONS--Areas of diagnostic difficulty mainly involve the subclassification of lymphomas. Review of tumour pathology by experts is recommended, at least in certain categories, to ensure correct diagnosis and uniformity in subclassification of tumours.
Improvement of gastric inflammation and resolution of epithelial damage one year after eradication of Helicobacter pylori.
Authors: Witteman E M , Mravunac M , Becx M J , Hopman W P , Verschoor J S , Tytgat G N , de Koning R W
Published: June 2007

AIMS--To investigate the effect of eradication of Helicobacter pylori infection on gastric epithelial damage and gastritis, scored according to the Sydney system. METHODS--Gastritis scores and epithelial damage were assessed in gastric biopsy specimens before, and five weeks and one year after anti-H pylori therapy in 66 patients with H pylori related gastritis. RESULTS--The mean initial levels of activity, inflammation, atrophy, intestinal metaplasia, and H pylori scores were higher in the antrum than in the corpus or fundus. Eradication of H pylori resulted in an improvement in the mean inflammatory score in antral biopsy specimens from 2.23 before treatment to 1.32 and 1.06, respectively, five weeks and one year after treatment. Corresponding values for fundic biopsy specimens were 1.30, 0.36 and 0.35. Activity scores improved from 1.41 before treatment to 0.13 and zero, respectively, five weeks and one year after treatment in antral biopsy specimens and from 0.60 before treatment to zero in fundic biopsy specimens. Before treatment, epithelial damage was present in 51% of biopsy specimens taken from the antrum and 23% of those from the corpus. Five weeks after eradication of H pylori none of the biopsy specimens revealed evidence of epithelial damage. CONCLUSION--Eradication of H pylori is followed by a rapid, significant improvement in the gastritis score and resolution of epithelial damage in antral and fundic mucosa.
Functional hyposplenism following allogeneic bone marrow transplantation.
Authors: Cuthbert R J , Iqbal A , Gates A , Toghill P J , Russell N H
Published: June 2007

AIMS--To investigate the incidence of functional hyposplenism in a group of patients who had undergone allogeneic bone marrow transplantation (BMT). METHODS--Splenic function was assessed by counting the number of gluteraldehyde fixed red blood cells containing pits or indentations as examined by interference phase microscopy. Normal values are < 2% whereas splenectomy patients have values of 25 to 40%. RESULTS--Twenty eight BMT recipients (17 men, 11 women) were studied at varying periods post-transplant and the results compared with 20 healthy volunteers and 10 patients who had undergone splenectomy or had splenic atrophy because of haematological conditions. Of the 28 BMT recipients, one had undergone a prior splenectomy; of the remaining 27 patients, four (15%) had evidence of functional hyposplenism with between 5.0 and 34.0% pitted cells. Of these four patients, one had active extensive chronic graft versus host disease (GvHD) which has been previously reported to be associated with functional hyposplenism following transplantation. Only one of the four patients had peripheral blood red cell changes typical of hyposplenism. CONCLUSION--These results confirm that extensive chronic GvHD is associated with hyposplenism. Intermediate degrees of functional hyposplenism may also occur following BMT in the absence of chronic GvHD and in the absence of haematological features of hyposplenism on routine blood films. This may be of significance in mediating the susceptibility to infection with encapsulating bacteria seen following allogeneic BMT.
Immunological factors and risk of infection in plateau phase myeloma.
Authors: Hargreaves R M , Lea J R , Griffiths H , Faux J A , Holt J M , Reid C , Bunch C , Lee M , Chapel H M
Published: June 2007

AIMS--A series of patients with myeloma were investigated to assess whether immunological risk factors predisposing to serious infection could be identified. METHODS--Patients (n = 102) with predominantly plateau phase myeloma were monitored prospectively for infections. Immunological parameters including total non-paraprotein immunoglobulins and specific antibody titres were measured in all patients and compared with a control population of healthy individuals of a similar age; response to immunisation with Pneumovax II, tetanus and diphtheria toxoids and IgG subclasses were measured in a subgroup of 41 patients. Other characteristics investigated for any association with infection included age, sex, paraprotein type, disease stage, and chemotherapy. RESULTS--Specific antibody titres to pneumococcal capsular polysaccharides and tetanus and diphtheria toxoids were significantly reduced compared with the control population. Low antipneumococcal and anti Escherichia coli titres correlated with risk of serious infection and low anti-pneumococcal titres with severity of non-paraprotein immunosuppression. In 41 immunised patients responses to Pneumovax II, tetanus and diphtheria toxoids were poor; IgG subclass levels were significantly reduced and a poor IgG response to Pneumovax II immunisation was associated with an increased risk of septicaemia and low IgG2 levels. The overall serious infection rate was 0.92 infections per patient year and was four times higher during periods of active disease (1.90) compared with plateau phase myeloma (0.49). The predominant site of infection was the respiratory tract. Clinical and laboratory parameters showed only male sex and reduced non-paraprotein IgG and IgA levels to be significantly associated with at least one serious infection. CONCLUSIONS--A subgroup of patients with myeloma with poor IgG responses to exogenous antigens, who are at increased risk of serious infection, can be identified and may benefit from replacement immunoglobulin therapy to reduce the risk of infection.
An unusual case of colonic angiodysplasia.
Authors: Trendell-Smith N J , Warren B F , Sheffield E A , Durdey P
Published: June 2007

An unusual case of a colonic vascular anomaly resembling angiodysplasia associated with right sided diverticular disease is presented. The patient, a 74 year old man, presented with a four day history of rectal bleeding and subsequently underwent hemicolectomy. The resected specimen was flushed out with heparin-saline solution and injected with a barium-gelatine mixture. Preoperative barium enema revealed right sided diverticula, whereas post-resection angioradiography revealed the "coral reef" vascular anomaly consistent with angiodysplasia. Histology confirmed the presence of both diverticular disease and angiodysplasia. This case report highlights the importance of considering a vascular anomaly in patients presenting with rectal bleeding despite the presence of another radiologically demonstrable anatomical lesion.
Evaluation of anticoagulant control in a pharmacist operated anticoagulant clinic.
Authors: Radley A S , Hall J , Farrow M , Carey P J
Published: June 2007

AIMS--To compare the quality of outpatient anticoagulant control before and after the transfer of dosing responsibility to designated trained pharmacists from rotating junior medical staff. METHODS--All International Normalised Ratio (INR) values for an eight month period either side of the staff changeover were assessed for precision of therapeutic control according to described standards. Allowing for patient associated effects, observed and expected frequencies of "successful" control for the two staff groups were compared under the hypothesis of no association. RESULTS--INR results (n = 2219) for 382 patients were analysed. For patients in stable therapeutic control, there was no significant difference in performance between the two staff groups. Patients with an INR result "out" of control limits were more likely to be returned "in" to control at their next visit by the pharmacists than by the doctors. CONCLUSIONS--The quality of anticoagulant control in outpatient clinics benefits from dedicated trained staff using standard protocols.
CD19 and immunophenotype of bone marrow plasma cells in monoclonal gammopathy of undetermined significance.
Authors: Zandecki M , Facon T , Bernardi F , Izydorczyk V , Dupond L , François M , Reade R , Iaru T , Bauters F , Cosson A
Published: June 2007

AIMS--To determine whether a particular phenotype or antigen is preferentially related to monoclonal gammopathies of undetermined significance (MGUS). METHODS--Bone marrow specimens from 56 patients with MGUS were stained immunocytochemically (ABC peroxidase) for CD38, CD56, CD9, CD10, CD19, CD20, CD22, and MB2. Specimens from patients recently diagnosed with multiple myeloma and reactive bone marrow samples were studied in parallel. RESULTS--CD38 was expressed on all plasma cells from all MGUS samples tested, while 36% were positive for CD56, CD9 and MB2 were both expressed strongly; CD20 was moderately expressed, and staining for CD10 and CD22 was uncommon. For these five B cell antigens there was no clear difference between their expression in MGUS and in multiple myeloma. A great difference was found for CD19: in MGUS this antigen was expressed on 2-91% of plasma cells (mean 35%) and 77% patients had > 10% positive plasma cells; in multiple myeloma its expression was low and only 12% patients had > 10% positive plasma cells. When these results were converted to numbers of CD19 positive plasma cells per 100 nucleated bone marrow cells, reactive bone marrow and MGUS specimens had a similar number of positive plasma cells. There was no correlation between expression of any of the antigens tested. CONCLUSIONS--Many of the so-called pre-B, B or activation antigens are present on plasma cells from MGUS specimens, and expression of CD9, CD10, CD20, CD22, MB2, and CD38 in MGUS was very similar to that in multiple myeloma. CD56 was frequently expressed in MGUS. In this series CD19 was highly expressed in MGUS but not in multiple myeloma. Plasma cells bearing this antigen could represent the non-neoplastic process and determination of its expression could be useful for the diagnosis of MGUS.
Internal audit in a microbiology laboratory.
Authors: Mifsud A J , Shafi M S
Published: June 2007

AIM--To set up a programme of internal laboratory audit in a medical microbiology laboratory. METHODS--A model of laboratory based process audit is described. Laboratory activities were examined in turn by specimen type. Standards were set using laboratory standard operating procedures; practice was observed using a purpose designed questionnaire and the data were analysed by computer; performance was assessed at laboratory audit meetings; and the audit circle was closed by re-auditing topics after an interval. RESULTS--Improvements in performance scores (objective measures) and in staff morale (subjective impression) were observed. CONCLUSIONS--This model of process audit could be applied, with amendments to take local practice into account, in any microbiology laboratory.
Detection of cytomegalovirus using PCR in serum from renal transplant recipients.
Authors: Cunningham R , Harris A , Frankton A , Irving W
Published: June 2007

AIMS--To develop a polymerase chain reaction (PCR) assay for the detection of cytomegalovirus (CMV) DNA in serum and leucocytes of renal transplant recipients and compare this assay with CMV culture and serodiagnosis. METHODS--Monthly specimens were obtained from 12 patients starting immediately before transplant. CMV infection was monitored by IgM enzyme linked immunosorbent assay, virus culture and PCR on serum and leucocytes. RESULTS--Two of four IgG positive patients had reactivation of CMV disease confirmed by culture, three of eight seronegative patients had a primary infection, one confirmed by serology and two by culture. PCR was positive earlier than conventional methods in three cases and concurrently in two. No positive PCR reactions occurred in the seven patients who remained negative by culture and serology. CONCLUSIONS--CMV DNA is detectable in serum; serum may be positive before virus is detectable by buffy coat culture; and PCR may be useful as an early indication of potential CMV disease in renal transplant recipients.
Lipoprotein composition and serum Lp(a) lipoprotein in hypobetalipoproteinaemia.
Authors: Crook M , Swaminathan R
Published: June 2007

A family with hypobetalipoproteinaemia was studied to examine the Lp(a) lipoprotein, lipoprotein cholesterol, and triglyceride composition of the serum lipids. Lp(a) lipoprotein was measured by immunoassay. Serum lipoproteins were separated by ultracentrifugation. Cholesterol and triglycerides were measured using standard enzymatic assays. Serum apolipoprotein B was low and Lp(a) undetectable in the index patient and in her father and son. Separation of the lipoproteins by ultracentrifugation showed a low cholesterol content of serum low density lipoprotein in the affected family members and also a low triglyceride content of high density lipoprotein particles in two affected members. It is concluded that serum lipoprotein cholesterol is altered in hypobetalipoproteinaemia, and family members of index cases have undetectable serum Lp(a) lipoprotein concentrations.
Malignant mesothelioma of the pleura: interobserver variability.
Authors: Andrion A , Magnani C , Betta P G , Donna A , Mollo F , Scelsi M , Bernardi P , Botta M , Terracini B
Published: June 2007

AIMS--To assess the consistency of histopathological diagnosis of pleural malignant mesothelioma by estimating interobserver agreement between five pathologists. METHODS--Eighty eight histological sets pertaining to original diagnoses of pleural malignant mesothelioma were reviewed separately by each pathologist. Diagnostic likelihood was graded as definite (A), probable (B), possible (C), improbable (D), and definitely not (E) malignant mesothelioma. The following indexes were estimated: observed proportion of agreement (Po), kappa statistics and proportion of agreement for "positive" (Ppos) and "negative" (Pneg) diagnoses. RESULTS--Sixty cases (68.2%) were rated by at least three reviewers as A or B and by none of the others as D or E. Five (5.7%) were rated by at least two reviewers as D or E and by none of the others as A or B. Nine (10.2%) showed a serious disagreement, rated A or B and D or E. Agreement for sets obtained at necropsy/surgery (median kappa w = 0.57) was similar to that for endoscopic material (median kappa w = 0.54). Agreement was poor on material obtained by needle biopsy (median kappa w = 0.21). The median value of Ppos varied between 0.94 (necropsy/surgery) and 0.67 (needle biopsy) and that of Pneg between 0.78 (necropsy/surgery) and 0.34 (unspecified biopsy). Interobserver agreement on histological typing was good overall (median kappa = 0.59). CONCLUSIONS--Of the original histological diagnoses, 70% were consistently reproduced through panel review. Most indexes of agreement between pathologists ranged from poor (needle biopsy) to moderate (necropsy/surgery). Agreement in confirming malignant mesothelioma was greater than that regarding exclusion of this diagnosis. Of the cases finally considered to have malignant mesothelioma, the reproducibility of histological typing was relatively satisfactory.
A "quickscore" method for immunohistochemical semiquantitation: validation for oestrogen receptor in breast carcinomas.
Authors: Detre S , Saclani Jotti G , Dowsett M
Published: June 2007

Immunohistochemistry is increasingly used in the assessment of markers for breast cancer prognosis. Semiquantitation is frequently desirable but, other than by the use of image analysis, the approaches currently in use are cumbersome. The most common method used is the H-score which takes into consideration the staining intensity in conjunction with the percentage of cells staining positively in breast carcinoma tissue. A "quickscore" has been developed which dispenses with the need to count individual cells. The quantitative biochemical Abbott enzyme immunoassay (EIA) and the Dako immunohistochemical assay (IHA) incorporating a semiquantitative H-score, have been used as standards against which the IHA quickscore for the semiquantitation of oestrogen receptor expression was tested. A good correlation was found between the quickscore and the EIA, which was as good as that between the H-score and EIA. The quickscore is a valid approach and there is no advantage in using the more rigorous H-score. A positive cut off quickscore of > or = 3 has been suggested.
Hepatitis C virus replication in hepatocellular carcinoma.
Authors: Niu J , Kumar U , Monjardino J , Goldin R , Rosin D , Thomas H C
Published: June 2007

Hepatitis C virus (HCV) replication is reported in both tumour and non-tumour tissue in a case of hepatocellular carcinoma. Viral replication was established by showing the presence of minus strand HCV RNA by PCR amplification, after excluding residual reverse transcriptase activity of Taq polymerase. No minus strand was found in serum derived virion RNA. PCR amplified products from both tumour and non-tumour parenchyma were sequenced in the 5' non-coding region and shown to be identical. The genotype of this Indonesian patient was found to be 1b (or II), the most prevalent type in the Far East.
Value of quantitative pathological variables as prognostic factors in advanced ovarian carcinoma.
Authors: Brinkhuis M , Baak J P , Meijer G A , van Diest P J , Mogensen O , Bichel P , Neijt J P
Published: June 2007

AIMS: To evaluate correlations among clinical, pathological, morphometric, stereological, and DNA flow cytometric variables and their prognostic value in advanced ovarian cancer. METHODS: Tissue was collected from 180 patients with advanced ovarian cancer. All 180 had undergone debulking surgery and were being treated with cisplatin. Long term follow up was available for all patients. The mitotic activity index (MAI), volume % of epithelium (VPE), mean nuclear area (MNA), standard deviation of the nuclear area (SDNA), estimates of volume weighted mean nuclear volume (nu v), and variables obtained from minimum spanning tree (MST) analysis were assessed in the least differentiated tumour section in each case. DNA flow cytometry was also performed. RESULTS: Quantitative pathological features differed significantly with respect to histological grade. The MAI, MNA, SDNA, and the number of points connected to three neighbours differed significantly among the different DNA ploidy groups. The VPE and number of points connected to two or three neighbours differed significantly between FIGO stages III and IV. Fifty two (29%) patients survived. FIGO stage, residual disease and SDNA had prognostic significance on both univariate and multivariate survival analysis. In patients with FIGO III stage disease and residual tumour nodes < or = 2 cm in diameter (67 patients, 29 (43%) survivors) a prognostic index was established based on SDNA and of the line length of the MST. The median survival time was not reached in a subgroup of patients with favourable prognosis (overall survival 57%). Median survival was 32 months for patients with an unfavourable index score (overall survival 28%). CONCLUSION: Morphometric variables have important additional value in predicting prognosis in patients with advanced ovarian cancer.
Blood ferritin concentrations in newborn infants and the sudden infant death syndrome.
Authors: Raha-Chowdhury R , Moore C A , Bradley D , Henley R , Worwood M
Published: June 2007

Liver iron concentrations have been shown to be higher in victims of SIDS than in postmortem controls suggesting that high levels of tissue iron may be implicated in SIDS. To determine whether infants who subsequently die from SIDS are born with greater iron stores than those who do not, the iron stores in newborn infants were assessed retrospectively by measuring blood ferritin concentration in spots from Guthrie cards (collected from almost all infants born in the UK in the first week of life). A method for extracting and measuring ferritin from stored blood spots is described. Eighteen cases of SIDS were identified in South Glamorgan along with four controls for each case. Ferritin concentrations did not differ in SIDS victims and controls suggesting that victims of SIDS are not born with abnormal concentrations of stored iron. If iron stores are found to be higher in SIDS victims than in healthy live infants of the same age then it is more likely that the iron will have been acquired after birth.
Chromosome 11q in sporadic colorectal carcinoma: patterns of allele loss and their significance for tumorigenesis.
Authors: Tomlinson I P , Bodmer W F
Published: June 2007

AIMS: To analyse the frequency of loss of heterozygosity (allele loss, LOH) in a large sample of colorectal carcinomas using highly informative markers along chromosome 11q. METHODS: One hundred paired samples of colorectal cancer and normal tissue were genotyped at six microsatellite markers on chromosome 11q (cen-D11S1313-D11S901-DRD2/NCAM-D11S29- D11S968-tel). The high levels of heterozygosity at these markers allow allele loss to be determined in about 80% of cases at any one locus. The frequency of replication errors (RERs, microsatellite instability) has also been determined. RESULTS: LOH was found at frequencies of 25% and 29% at the distal D11S968 (11qter) and D11S29 (11q23.3) loci, slightly above the accepted baseline of 0-20%. Allele loss at NCAM, DRD2, D11S901, and D11S1313 was not raised above baseline levels. The probable genetic mechanism of allele loss--chromosomal non-disjunction, mitotic recombination, deletion, or gene conversion--seemed to vary between tumours and no consistent mechanism of mutation was found. Microsatellite instability was found in 23 (23%) tumours. No associations were found between LOH and clinical data (patient sex, age at presentation, tumour site, and Duke's stage). CONCLUSIONS: Although gene(s) on 11q may have a role in the development of a minority of colorectal carcinomas, this study provides evidence against the general importance of allele loss on chromosome 11q in the pathogenesis of colorectal cancer. The results also have implications for the importance of 11q in other cancers: it seems less likely that a single tumour supressor gene at this location promotes the growth of all types of tumour when lost. Rather, one or more genes with tissue specific effects may be involved.
Alpha-fetoprotein production by a hepatoid adenocarcinoma of the uterus.
Authors: Yamamoto R , Ishikura H , Azuma M , Hareyama H , Makinoda S , Koyama Y , Nishi S , Fujimoto S
Published: June 2007

A case of a 62 year old Japanese woman with an endometrial adenocarcinoma producing alpha-fetoprotein (AFP) is described. Microscopically, the tumour was composed of a major medullary portion and a minor tubular adenocarcinoma which had invaded the myometrium, the myometrial lymphatics and blood vessels. Neoplastic cells in the medullary portion were polygonal with glycogen-rich cytoplasm. Vascular permeation by neoplastic cells was prominent. Extensive hepatoma-like features were observed. The tumour cells lacked features suggestive of a diagnosis of embryonal carcinoma or endodermal sinus tumour. The production of AFP by the tumour cells was demonstrated immunohistochemically using the PAP technique. Only two cases of AFP producing endometrial adenocarcinomas have been reported previously.
Differential regulation of leucocyte L-selectin (CD62L) expression in normal lymphoid and inflamed extralymphoid tissues.
Authors: Munro J M , Briscoe D M , Tedder T F
Published: June 2007

AIMS: To study tissue expression of L-selectin, a leucocyte cell surface molecule that is considered to be involved in adhesion to certain endothelia, particularly in peripheral lymph nodes and during inflammation, and is shed upon leucocyte activation. METHODS: Leucocytes were examined by immunohistochemistry and double immunofluorescence staining in various lymphoid sites and normal and inflamed extralymphoid tissues. RESULTS: L-selectin was present on mantle zone B lymphocytes in different lymphoid sites, including in intestinal lymphoid tissue, but was absent on germinal centre B cells. Splenic white pulp B cells also expressed L-selectin. The proportion of T lymphocytes expressing L-selectin depended on the site under study, being greatest in peripheral lymph nodes (mean 48% of T cells positive), and lower in mucosal lymphoid sites and spleen (9 and 11% positive, respectively). Non-lymphocytic L-selectin staining was observed on follicular dendritic cells in tonsils and on macrophages in thymus. L-selectin positive leucocytes were rare in normal extralymphoid tissues, and relatively few were seen in most inflammatory settings. However, in rejecting renal transplants, a higher proportion (30%) of leucocytes expressed L-selectin. CONCLUSIONS: Overall, the results indicate how the degree of L-selectin expression by leucocytes in particular tissues may reflect a requirement for L-selectin expression for entry into those tissues and the activation state of leucocytes once localised there.
Immunohistological analysis of tumour growth factor beta 1 expression in normal and inflamed salivary glands.
Authors: Kizu Y , Sakurai H , Katagiri S , Shinozaki N , Ono M , Tsubota K , Saito J
Published: June 2007

AIM: To determine whether transforming growth factor-beta 1 (TGF-beta 1) has a pathogenetic role in disease of the salivary glands. METHODS: An indirect immunohistochemical technique was used to analyse TGF-beta 1 expression in six specimens of normal salivary gland and 23 surgical specimens. RESULTS: TGF-beta 1 was strongly expressed in the ductal epithelial cells of normal salivary gland tissues (six of six cases) and in inflammatory conditions (eight of 11 cases). In contrast, TGF-beta 1 was not detectable in ductal epithelial cells expressing HLA-DR around infiltrating CD4+ CD45RO+ activated T cells, in the salivary gland tissue of patients with Sjögren's syndrome. CONCLUSION: Because TGF-beta 1 has an essential role in the mucosal immunity of salivary glands, abnormal expression of this cytokine must be regarded as a candidate in the pathogenesis of Sjögren's syndrome.
The coroner's necropsy in sudden death: an under-used source of epidemiological information.
Authors: O'Sullivan J P
Published: June 2007

AIMS: To determine the number of unsuspected disease processes found in a series of cases of sudden unexpected death occurring outside hospital and to enumerate how many of these were not recorded on the death certificate. METHODS: In a series of 1000 routine coroners' necropsies for sudden unexpected death, major findings that had not been known about in life were recorded. Macroscopic findings were confirmed histologically as appropriate. The deaths occurred either outside hospital or in the Accident and Emergency department before the patient could be examined. Cot deaths and decomposed bodies were excluded. RESULTS: There were 575 major findings in 532 (53.2%) subjects that had been clinically silent in life. Of these 575 findings, 277 (48.2%) were not the cause of death and so did not appear on the death certificate. Eighty per cent of the major alimentary system findings and all of the genitourinary findings were of this type. In addition, however, 30% of the major cardiovascular and 34% of the major respiratory findings were not recorded on the death certificate for this reason. CONCLUSIONS: A large amount of important epidemiological data is being lost in the operation of the coronial system. Some of this information is irrecoverable as the function of the death certificate is to provide a cause of death only. In addition, information may be being lost because the necropsy is not being performed adequately and is not subject to audit.
Use of methyl methacrylate resin for embedding bone marrow trephine biopsy specimens.
Authors: Blythe D , Hand N M , Jackson P , Barrans S L , Bradbury R D , Jack A S
Published: June 2007

AIMS: To evaluate the use of methyl methacrylate resin as an embedding medium for undecalcified bone marrow trephine biopsy specimens. METHODS: About 2500 undecalcified bone marrow trephine biopsy specimens were processed, and embedded in methyl methacrylate resin. Semithin sections (2-3 microns) were stained by routine tinctorial and immunocytochemical staining methods with a wide range of antibodies using a standard streptavidin biotin horseradish peroxidase technique. Different antigen retrieval pretreatments were evaluated. RESULTS: Bone marrow trephine biopsy specimens are embedded routinely in methyl methacrylate at the Haematological Malignancy Diagnostic Service at The Leeds General Infirmary. Over 50 different primary antibodies are in current use; for the majority of these, microwave antigen retrieval or trypsin digestion, or both, is either essential or greatly enhances the results. CONCLUSIONS: Embedding bone marrow trephine biopsy specimens in methyl methacrylate resin retains morphology and permits reliable, high quality immunocytochemistry. This is particularly desirable for the demonstration of neoplastic cells in regenerative marrow after chemotherapy, and in the detection of residual disease after treatment. The use of methyl methacrylate for routine use on bone marrow trephine biopsy specimens is advocated.
Angiotropic lymphoma: report of a case with histiocytic features.
Authors: Snowden J A , Angel C A , Winfield D A , Pringle J H , West K P
Published: June 2007

Angiotropic lymphoma, also known as intravascular lymphomatosis, is characterised by widespread intravascular proliferation of malignant lymphoid cells, usually without evidence of focal disease. A case of a 52 year old man referred for investigation of a two year history of pyrexia of unknown origin, skin rash and multiple organ failure is described. Angiotropic lymphoma was seen in gastric, colonic and skin biopsy specimens, and review of an earlier skin biopsy specimen showed similar morphological features. In contrast to previous cases which showed B or T cell differentiation, immunohistochemical examination was positive for histiocyte markers. Molecular studies showed no evidence of immunoglobulin heavy chain gene or T cell receptor gene rearrangement. The patient responded to combination chemotherapy, comprising cyclophosphamide, doxorubicin, etoposide, and prednisolone. This case highlights the fact that advanced lymphoma may be present without evidence of focal disease and that the diagnosis may be missed easily both clinically and histologically.