bluuebunny/crossref_metadata_2025 huggingface dataset:
Use of PCR in resolving diagnostic difficulties potentially caused by genetic variation of hepatitis B virus.
Authors: van Deursen F J , Hino K , Wyatt D , Molyneaux P , Yates P , Wallace L A , Dow B C , Carman W F
Published: January 2009

AIMS: To assess the relevance of genetic variants of hepatitis B virus (HBV) and to demonstrate the usefulness of the polymerase chain reaction (PCR) in cases of HBV diagnostic difficulty. METHODS: Five serum samples from patients that presented diagnostic difficulty in routine laboratories were sent to a research laboratory for PCR, and if appropriate, S gene sequencing, in vitro expression, and antigenic analysis. RESULTS: The demonstration of HBV in serum by PCR allowed a definitive diagnosis of current infection. One serum sample with poor reactivity in a diagnostic assay had a minor hepatitis B surface antigen (HBsAg) variant and another with very poor reactivity had multiple variants of HBsAg. Transient HBsAg reactivity was observed in a recently vaccinated patient. A hepatitis Be antigen (HBeAg) false positive reaction was noted in a patient from a well defined risk group for HBV. One patient who was strongly HBsAg/HBeAg positive, but anti-hepatitis B core antibody negative, was viraemic. CONCLUSIONS: PCR may become the gold standard for the diagnosis of current HBV infection. HBV variants are responsible for a proportion of diagnostically difficult cases. Modification of commercial assays is necessary to increase the sensitivity of detection of such variants.
Differentiation between actinic reticuloid and cutaneous T cell lymphoma by T cell receptor gamma gene rearrangement analysis and immunophenotyping.
Authors: Bakels V , van Oostveen J W , Preesman A H , Meijer C J , Willemze R
Published: January 2009

AIMS: Differentiation between actinic reticuloid and cutaneous T cell lymphoma can be extremely difficult. Demonstration of clonal T cell receptor (TCR) gene rearrangements has been suggested as a potential diagnostic criterion, but the results obtained thus far have been conflicting. This study investigated whether TCR gamma gene rearrangement analysis, using polymerase chain reaction (PCR) in combination with denaturing gradient gel electrophoresis (DGGE) and immunohistochemistry, can serve as a diagnostic criterion. METHODS: PCR/DGGE was performed on skin, peripheral blood mononuclear cells, and/or lymph nodes of seven patients with actinic reticuloid, 11 patients with Sézary syndrome, and 15 patients with a benign form of erythroderma. The results of PCR/DGGE and Southern blot analysis of TCR beta gene rearrangements were compared. In addition, CD4:CD8 ratios in skin and peripheral blood samples were investigated. RESULTS: Clonal T cell populations were detected in 19 of 21 samples obtained from patients with Sézary syndrome but were not detected in any of the 12 samples from patients with actinic reticuloid. Clonal T cells were detected in the peripheral blood of only one of 15 patients with a benign form of erythroderma. PCR/DGGE and Southern blot analysis gave concordant results in 28 of 29 samples. Immunophenotypic analysis demonstrated increased proportions of CD8+ T cells in skin (seven of seven cases) and peripheral blood (four of seven cases) of patients with actinic reticuloid. CONCLUSION: The results of this study demonstrate that gene rearrangement analysis, in combination with immunohistochemistry, may be an important adjunct in differentiating between actinic reticuloid and cutaneous T cell lymphoma. In patients suspected of having actinic reticuloid, application of both techniques is recommended.
p21WAF1/CIP1 expression in gestational trophoblastic disease: correlation with clinicopathological parameters, and Ki67 and p53 gene expression.
Authors: Cheung A N , Shen D H , Khoo U S , Wong L C , Ngan H Y
Published: January 2009

BACKGROUND: The p21WAF1/CIP1 gene mediates growth arrest by inhibiting G1 cyclin dependent kinases and has been considered as a downstream effector of the tumour suppressor gene p53. AIM: To analyse the role of p21WAF1/CIP1 in gestational trophoblastic disease. METHODS: The immunohistochemical expression of p21WAF1/CIP1 gene was measured in 33 placentas, 28 partial hydatidiform moles, 54 complete hydatidiform moles, and 13 choriocarcinomas in paraffin wax embedded tissue. The results were correlated with p53 (DO7) and Ki67 (MIB1) immunoreactivity as well as clinical progress. RESULTS: p21WAF1/CIP1 immunoreactivity was found predominantly in the nuclei of the syncytiotrophoblasts. p21WAF1/CIP1 protein expression correlated with gestational age in normal placentas (p = 0.0001) but not in hydatidiform moles (p = 0.89). Complete hydatidiform moles and choriocarcinomas had a significantly higher p21WAF1/CIP1 expression compared with normal placentas and partial hydatidiform moles (p < 0.001); there was no difference between placentas and partial hydatidiform moles. No correlation between p21WAF1/CIP1 expression and either the proliferation (Ki67) index (p = 0.34) or p53 protein accumulation (p = 0.68) was demonstrated. There was no significant difference (p > 0.05) in p21WAF1/CIP1 expression between the 17 patients who developed persistent gestational trophoblastic disease and those who did not. CONCLUSIONS: This study suggests that p21WAF1/CIP1 expression in trophoblastic disease may be induced by a p53 independent pathway. The proliferative activity of gestational trophoblastic diseases might not be determined solely by the control of the cell cycle operated by p21WAF1/CIP1. p21WAF1/CIP1 expression is not an accurate prognostic indicator of gestational trophoblastic disease.
How many lymph nodes to stage colorectal carcinoma?
Authors: Mainprize K S , Kulacoglu H , Hewavisinthe J , Savage A , Mortensen N , Warren B F
Published: January 2009

This study aimed to establish the number of cassettes that should be filled with lymph nodes to stage a colorectal carcinoma as Dukes's stage C. The records from the Oxford Colorectal Cancer database of all patients diagnosed with Dukes's stage C cancer from late 1988 to early 1993 were reviewed. Each slide of lymph nodes was examined to determine how many slides needed to be looked at to find the first positive lymph node. The resected specimens were not fat cleared but dissected manually in a routine fashion. One hundred and eight slides were retrieved. The mean total lymph node harvest was 8.44 for each patient. Ninety eight patients (90.7%) had positive lymph nodes on the first slide with an average of 3.42 lymph nodes on each slide, of which a mean of 1.82 were positive. For nine patients, two slides were required to make a diagnosis of lymph node involvement, and for one patient the first three slides needed inspection to establish Dukes's stage C. In conclusion, using a routine technique to obtain lymph nodes from colorectal cancer specimens 99% of Dukes's stage C colorectal carcinoma can be found by filling two cassettes with lymph nodes.
Increased expression of proliferating cell nuclear antigen in autoimmune hepatitis in a patient with raised serum concentration of CA19-9.
Authors: Sohda T , Iwata Y , Shijo H , Egashira Y , Egashira K , Okumura M
Published: January 2009

A 52 year old woman had autoimmune hepatitis and an increased concentration of serum carbohydrate antigen 19-9 (CA19-9). The origin of the raised CA19-9 was studied using immunohistochemistry. Liver biopsy section showed chronic active hepatitis with large numbers of proliferated bile ductules. Immunohistochemical analysis revealed that the proliferated bile ductule cells were positive for proliferating cell nuclear antigen (PCNA) and for CA19-9. It is speculated that the raised serum CA19-9 concentration was derived from proliferated bile ductule cells and these cells, which are positive for PCNA, may be able to produce high concentrations of CA19-9.
A case of systemic pseudo-pseudoxanthoma elasticum with diverse symptomatology caused by long-term penicillamine use.
Authors: Coatesworth A P , Darnton S J , Green R M , Cayton R M , Antonakopoulos G N
Published: January 2009

A 47 year old man presented with a two year history of increasing cervical dysphagia, dyspnoea, and cutaneous signs. He had been diagnosed 27 years previously with Wilson's disease and was treated with penicillamine (1.5 g daily). Systemic abnormality of elastic fibres was confirmed by light and electron microscopy following biopsy of skin, lung, oesophageal muscle, gum, pharyngeal tissue, and cervical connective tissue. Dysphagia was relieved by cricopharyngeal myotomy. Substitution of trientene dihydrochloride for penicillamine relieved cutaneous and systemic manifestations. This is possibly the first case demonstrating an association between prolonged penicillamine use and biopsy proved systemic pseudo-pseudoxanthoma elasticum. The presenting symptoms may have resulted from the abnormal numbers and properties of elastic fibres, and the changes were caused by penicillamine use, rather than by idiopathic, inherited pseudoxanthoma elasticum.
Intravascular synovial sarcoma.
Authors: Robertson N J , Halawa M H , Smith M E
Published: January 2009

A case of intravascular biphasic synovial sarcoma arising from the wall of the left femoral vein in a 34 year old woman is described. This is the third case of an intravascular synovial sarcoma known to be reported in the medical literature. The two previous cases arose from the left femoral vein and inferior vena cava in women of 34 and 31 years old, respectively. A characteristic clinical pattern appears to be emerging--that is, location in large veins of the lower extremity and trunk in young adult females. Synovial sarcoma must be considered in the differential diagnosis of intravascular "tumours".
Autografting as first line treatment for chronic myeloid leukaemia.
Authors: Singer I O , Franklin I M
Published: January 2009

Interest in autografting for chronic myeloid leukaemia and its clinical relevance has revived in recent years. This followed observations that with various chemotherapeutic regimens it was possible to achieve, temporarily at least, peripheral blood and bone marrow that were Philadelphia negative. Bone marrow or peripheral blood progenitor cells could then be harvested and reinfused following a high dose procedure, hopefully eliminating any residual disease, and resulting in prolonged disease free survival. This ideal has not yet been successfully achieved with current strategies. Recent results indicate that eliminating residual disease with current chemotherapy is not normally achievable. The use of more sensitive technologies such as polymerase chain reaction has revealed persistent disease in most if not all apparently Philadelphia negative cases. This is confirmed by results where disease relapse occurs following transplant in these cases. Despite this, clinically relevant remissions are obtained and further trials are indicated. In this review present treatment is discussed and future strategies, using novel techniques as an adjunct to current treatment, are proposed that might improve on present results or even lead to the elusive goal of cure.
Aetiology, pathogenesis, and pathology of cervical neoplasia.
Authors: Arends M J , Buckley C H , Wells M
Published: January 2009

Early epidemiological studies of cervical neoplasia suggested a causal relation with sexual activity and human papillomaviruses (HPVs) have emerged as prime suspects as venerally transmitted carcinogens. HPVs fall into two broad camps: low risk types, associated with cervical condylomas and CIN 1; and high risk types (mostly 16 and 18), found in 50-80% of CIN 2 and CIN 3 lesions, and 90% of cancers. This association with cancer is very strong, with odds ratios of > 15 (often much higher) in case-control studies that are methodologically sound. An infrequently detected third group of intermediate risk type HPVs is associated with all grades of CIN and occasionally with cancers. HPVs have also been detected in a wide range of asymptomatic controls, indicating that other events are required for development of neoplasia such as viral persistence and/or altered expression of viral genes, often following integration of the viral genome. This leaves the two major viral oncogenes, E6 and E7, directly coupled to viral enhancers and promoters, allowing their continued expression after integration. High risk HPV E7 proteins bind and inactivate the Rb protein, whereas E6 proteins bind p53 and direct its rapid degradation. A range of putative cofactors has been implicated in progression: HLA type, immunosuppression, sex steroid hormones, and smoking; most of these cofactors appear to influence progression to CIN 3. The natural history includes progression to CIN 3 in 10% of CIN 1 and 20% of CIN 2 cases, whereas at least 12% of CIN 3 cases progress to invasive carcinoma. Cervical glandular intraepithelial neoplasia (CGIN) often coexists with squamous CIN, and the premalignant potential of high grade CGIN is not in doubt, but the natural history of low grade CGIN remains uncertain. A high proportion of CGIN lesions and adenocarcinomas are HPV positive, and HPV18 has been implicated more in glandular than in squamous lesions. A strong clinical case for the application of HPV typing of cells recovered from cervical scrapes can be made; however, a rigorous cost-benefit analysis of introducing HPV typing into the cervical screening programme is required. Prophylactic and therapeutic HPV vaccines are under development. This article reviews the aetiology, pathogenesis, and pathology of cervical neoplasia, emphasising the role of HPVs.
ACP Broadsheet No 152: March 1998. Clinical implications of plasma homocysteine measurement in cardiovascular disease.
Authors: Still R A , McDowell I F
Published: January 2009

Plasma homocysteine is emerging as an important risk factor for cardiovascular disease. Measurement in clinical laboratories is feasible by several techniques, including HPLC. Of particular importance is the potential for treatment by dietary modification and vitamin supplements, particularly folic acid. The outcome of vitamin intervention on cardiovascular events is awaited with interest. If the role of homocysteine is confirmed then it may rival cholesterol in importance as a cardiovascular risk factor.
Pseudoangiomatous hyperplasia of mammary stroma associated with gynaecomastia.
Authors: Milanezi M F , Saggioro F P , Zanati S G , Bazan R , Schmitt F C
Published: January 2009

AIMS: To evaluate the prevalence of pseudoangiomatous hyperplasia of mammary stroma in gynaecomastia and its immunohistochemical profile in this setting. METHODS: Eighty eight cases of gynaecomastia recovered from the files of the department of pathology, Botucatu School of Medicine from 1976 to 1996 were studied. In the cases associated with pseudoangiomatous hyperplasia of mammary stroma, immunoreactivity for cytokeratins (CAM 5.2), vimentin, CD34, factor VIII related antigen, and the oestrogen and progesterone receptors were studied. RESULTS: Pseudoangiomatous hyperplasia of mammary stroma was found in 21 of 88 cases of gynaecomastia (23.8%). In all cases, the cells lining the spaces were positive for vimentin, whereas CAM 5.2 and factor VIII related antigen were consistently negative. Nineteen of the 21 cases showed immunoreactivity for CD34. Ductal epithelial cells were positive for both the oestrogen receptor and the progesterone receptor, whereas stromal cells were negative. CONCLUSIONS: Pseudoangiomatous hyperplasia of mammary stroma was present in approximately one quarter of the cases of gynaecomastia. This immunohistochemical study confirms the mesenchymal origin of the stromal cells that line the pseudovascular spaces, as has been found in female cases of pseudoangiomatous hyperplasia of mammary stroma.
Quantitative histopathology can aid diagnosis in painful bladder syndrome.
Authors: Thilagarajah R , Witherow R O , Walker M M
Published: January 2009

AIMS: To define the pathology of painful bladder syndrome using a morphometric method. METHODS: Bladder biopsy specimens from 31 patients with painful bladder syndrome and 11 controls were stained and examined at x260 magnification with the aid of a 100 square counting grid. Random counts of the different tissues and inflammatory components were made to ascertain whether constant differences occurred that could be used to define the pathology of this uncommon condition. RESULTS: In the lamina propria of painful bladder syndrome specimens, a significant increase was seen in the concentration of lymphocytes, T cells, and blood vessels; a decrease was seen in the number of fibroblasts, and no change was seen in the number of mast cells and macrophages. B cells were sporadic. The basement membrane in these specimens showed significant discontinuity and there was increased collagen deposition in the underlying muscle when compared with controls. CONCLUSION: Painful bladder syndrome exhibits constant histological features that may be used to aid diagnosis in this uncommon condition. Simple numerical cell/tissue measurement of this kind is also useful when treatment trials are considered, because objective statistical analysis (pretreatment and post-treatment) is possible without the need for expensive and complicated equipment.
Evaluation of the efficacy of 99mTc-Infecton, a novel agent for detecting sites of infection.
Authors: Hall A V , Solanki K K , Vinjamuri S , Britton K E , Das S S
Published: January 2009

AIMS: To determine the sensitivity and specificity of 99mTc-Infecton (Infecton), a novel ciprofloxacin based imaging agent, in detecting sites of infection. METHODS: Ninety patients thought to be suffering from a variety of infections were administered 300-400 MBq of Infecton intravenously. Whole body images were taken one and four hours later. Appropriate specimens were taken for microbiological investigations. Statistical analysis was performed using a computer statistical package. RESULTS: Ninety eight Infecton images were produced. Forty one of these were positive, including three false positives, where the patients had non-infective conditions. Fifty seven negative images were obtained, of which 41 were true negatives and 16 were false negatives, having definite evidence of infection. Thus, Infecton imaging has a sensitivity of 70.3% and a specificity of 93.1% for detecting infective foci. The positive and negative predictive values were 92.6% and 71.9%, respectively. CONCLUSION: Infecton imaging is a new diagnostic tool that is specific for detecting sites of bacterial infection in the body. The high positive predictive value displayed by the technique is clinically important because a positive image strongly supports a diagnosis of bacterial infection. A negative result does not rule out an infection, and may be a result of previous or current antibiotic treatment and/or infection with organisms that do not take up Infecton. Infecton imaging has major advantages over well established imaging techniques, including radiolabelled leucocytes, and may prove to be a superior method for localising bacterial infections.
Omeprazole may exert both a bacteriostatic and a bacteriocidal effect on the growth of Helicobacter pylori (NCTC 11637) in vitro by inhibiting bacterial urease activity.
Authors: Mirshahi F , Fowler G , Patel A , Shaw G
Published: January 2009

AIMS: To assess the potential antibacterial effect of omeprazole, a benzimidazole proton pump inhibitor, on the growth of Helicobacter pylori in vitro and to evaluate the effect of this compound on bacterial urease activity. METHODS: The growth of H pylori was observed in liquid culture in the presence and absence of omeprazole (0.8 mg/ml). Urease activity was evaluated in aliquots removed from two hour cultures by monitoring the initial change in absorbency at 560 nm in the presence of 0.02% phenol red. RESULTS: The minimum inhibitory concentration of omeprazole against H pylori was 0.8 mg/ml. The concentration of omeprazole required to inhibit growth was dependent on inoculum density: omeprazole (0.8 mg/ml) prevented growth from a 1 x 10(6) cfu/ml inoculum, but not from the higher inocula of 10(7) or 10(8) cfu/ml. This is the first study to demonstrate that omeprazole exerts a bacteriocidal effect against low bacterial densities and a bacteriostatic effect when bacterial density is high. When used at the onset of growth, this concentration of omeprazole has a bacteriocidal effect after four hours, although it exerts a bacteriostatic effect when added to cultures after the exponential phase. Bacterial urease activity is competitively inhibited by omeprazole in a dose dependent manner. CONCLUSION: The results suggest that omeprazole exerts both a bacteriocidal and a bacteriostatic effect against H pylori and competitively inhibits bacterial extracellular urease activity.
Effect of kidney function and disease status on urinary tissue factor measurements.
Authors: Lwaleed B A , Bass P S , Rogerson M E , Francis J L , Chisholm M
Published: January 2009

AIM: To investigate factors that influence urinary tissue factor (uTF) measurements: glomerular permeability and filtration, tubular function, haematuria, and urine bacterial growth. METHODS: uTF, protein creatinine index, glomerular filtration rate, retinol binding protein, N-acetyl-beta-D-glucosaminidase (NAG) and urinary haemoglobin (uHb) were measured in patients with hypertension, diabetes mellitus and nephrotic syndrome (n = 342), tubulo-interstitial disease (n = 50), and haematuria of uncertain cause (n = 50); measurements were also made in urine samples from healthy subjects for "simulated" haematuria (n = 6) and bacterial growth (n = 4) studies. RESULTS: There was a weak correlation of uTF with glomerular permeability and filtration (protein creatinine index and glomerular filtration rate) and with markers of tubular function (retinol binding protein and NAG). uTF concentrations were not affected by the presence of blood or bacteria in the urine sample. CONCLUSION: uTF concentrations are relatively stable. This is an important finding if the assay is to be used in clinical practice.
Antimony in blood and urine of infants.
Authors: Cullen A , Kiberd B , Matthews T , Mayne P , Delves H T , O'Regan M
Published: January 2009

AIM: To establish a reference range for antimony in the serum and urine of infants in the first year of life. METHODS: 100 infants were selected randomly from the population. Each infant had a single blood and urine sample taken. Antimony was assayed using inductively coupled plasma mass spectrometry. RESULTS: The reference range for antimony in the serum of infants in the first year of life was established as 0.09-0.25 microgram/l. The upper 95% centile for urinary antimony, corrected for creatinine, in the same population was 2.6 ng/mg creatinine. There was a very weak correlation between the serum and urine concentrations. CONCLUSIONS: This study confirms the presence of low concentrations of antimony in the serum and urine of healthy infants.
A pseudo-cryptococcal artefact derived from leucocytes in wet India ink mounts of centrifuged cerebrospinal fluid.
Authors: Thiruchelvan N , Wuu K Y , Arseculeratne S N , Ashraful-Haq J
Published: January 2009

Wet India ink mounts of cerebrospinal fluid (CSF) are useful in the laboratory diagnosis of cryptococcal meningitis. Pseudo-cryptococcal artefacts in such mounts have been attributed to leucocytes in CSF but their mode of formation has not been explained. This report describes the reproduction of such an artefact in cryptococcus free CSF-leucocyte mixtures that had been subjected to high speed centrifugation. The viscosity of DNA that could provide a morphological pseudo-capsule, and the yellow-green fluorescence of the pseudo-capsular material on staining with acridine-orange, suggest that lymphocytic nuclear DNA, which possibly leaked out after damage to the lymphocyte membrane by centrifugation, was responsible for this artefact.
Unacceptably high site variability in postmortem blood alcohol analysis.
Authors: Sylvester P A , Wong N A , Warren B F , Ranson D L
Published: January 2009

Blood alcohol concentration is a frequently requested test in forensic pathology. The variability of this value was studied by measuring the blood alcohol concentration from six sites in nine subjects at necropsy in whom alcohol was the implicated cause of death. There were small consistent differences in the blood alcohol concentrations between the sites in the nine subjects (p < 0.04). Calculation of the mean blood:vitreous humour alcohol concentration ratio (B:V ratio) showed that vitreous humour alcohol concentration most closely reflected the concentration at the femoral vein (B:V ratio = 0.94, r = 0.98), which is considered the optimal site for blood alcohol measurement. The correlation of left heart blood with femoral blood was lower compared with the other sites. There is a potential for an unacceptably large variation in the postmortem measurement of blood alcohol within each subject.
Value of assessing parathyroid hormone-like activity in a case of extreme hypercalcaemia.
Authors: Ranganath L , Jamal H , Jones L , Goddard P F
Published: January 2009

A previously well 70 year old woman was admitted to hospital following a three day history of vomiting and confusion. Her serum calcium was 6.58 mmol/l, phosphate 1.09 mmol/l, and alkaline phosphatase 91 iu/l. The mechanism of this hypercalcaemia was not obvious as there was no evidence of a primary malignancy, lymphadenopathy or hepatosplenomegaly. The calculation of indices of urinary excretion of calcium and phosphate suggested the presence of excessive parathyroid hormone (PTH) activity as the mechanism of hypercalcaemia. Plasma intact PTH, 25-hydroxycholecalciferol, and 1,25-dihydroxycholecalciferol were not raised suggesting the presence of PTH related peptide (rP). This led to a systematic search for a malignancy, which revealed the presence of a high grade B cell non-Hodgkin's lymphoma confined to the bone marrow. Plasma PTH-rP was subsequently shown to be raised confirming the interpretation of the initial urinary and calcium excretion indices. This case highlights the value of standard laboratory measurements such as urinary calcium and phosphate excretion in cases of hypercalcaemia of obscure aetiology, which can complement measurements of PTH and other calcitropic hormones.
Application of molecular typing to the epidemiology of Streptococcus pneumoniae.
Authors: Hall L M
Published: January 2009

The spread of antibiotic resistance and the development of new vaccines have focused attention on the epidemiology of Streptococcus pneumoniae over recent years. While serotyping and the determination of antibiotic resistance remain primary methods for characterising pneumococci, molecular typing can add greater discrimination and complementary information. Methods based on restriction fragment length polymorphism within total DNA or non-specific polymerase chain reaction provide information representative of the whole genome and can be used to recognise closely related isolates from different sources, whether in the investigation of possible cross infection at the local level or in the investigation of national or international spread of antibiotic resistant strains. Fingerprinting of penicillin binding protein genes adds further information in the analysis of penicillin resistant isolates. The use of a combination of typing methods to analyse both the genome as a whole and specific loci has led to the realisation that pneumococci undergo horizontal gene transfer much more often than most other bacterial species. In particular the spread of penicillin resistance has been characterised by a combination of the spread of epidemic strains, transfer of chromosomal resistance genes from such strains into other genetic backgrounds, and transfer of capsule genes resulting in the switch of serotypes within strains. In the future molecular typing will have an important role in discovering whether widespread vaccination leads to genetic modification of the pneumococcal population causing invasive disease.
Genetic polymorphism of N-acetyltransferase-2, glutathione S-transferase-M1, and cytochromes P450IIE1 and P450IID6 in the susceptibility to head and neck cancer.
Authors: González M V , Alvarez V , Pello M F , Menéndez M J , Suárez C , Coto E
Published: January 2009

AIMS: To analyse the allele frequencies of DNA polymorphisms at the genes for cytochromes P450IIE1 and P450IID6, N-acetyltransferase-2, and glutathione S-transferase-M1 in patients with head and neck squamous cell carcinoma, in an attempt to define genetic factors involved in the susceptibility to this cancer, which is strongly associated with tobacco consumption. METHODS: Determination of restriction fragment length polymorphism (RFLP) at cytochromes P450IIE1/P450IID6 and NAT2 genes, and the presence of homozygous deletion of the GSTM1 gene, in 200 controls and 75 head and neck cancer patients. Allelic frequencies between the two groups were compared using a chi 2 test, and odds ratio with 95% confidence intervals were calculated. RESULTS: There was no evidence of an association between alleles of CYP2D6 and CYP2E1 and head and neck cancer in our population. Similarly, frequencies of individuals lacking the GSTM1 gene did not differ between controls and patients. However, individuals with the NAT2-SA phenotype were at higher risk of developing head and neck cancer. The frequencies of the most common SA genotype (homozygous for the NAT2*5 allele) were higher in patients than in controls (27% v 15%, respectively). Slow acetylators homozygous for the NAT2*6 allele, the second most common SA allele, were also more common in patients than in controls (11% v 5%, respectively). CONCLUSIONS: Slow NAT2 activity is a risk factor possibly leading to the development of head and neck cancer in response to tobacco carcinogens.
Evaluation of Helicobacter pylori vacA genotype in Japanese patients with gastric cancer.
Authors: Shimoyama T , Yoshimura T , Mikami T , Fukuda S , Crabtree J E , Munakata A
Published: January 2009

AIM: To examine the vacA genotypes of Helicobacter pylori strains in Japan and to define whether any specific genotype was associated with gastric cancer. METHODS: The allelic variation of vacA gene was studied using a recently introduced polymerase chain reaction based vacA genotyping system. RESULTS: 80 H pylori strains were isolated from gastric biopsies of 40 patients with gastric cancer and 40 control subjects in a Japanese population. All strains were s1/m1 subtype and 79 of 80 strains were classified as s1a subtype. CONCLUSIONS: The recently proposed vacA genotyping system is applicable to Japanese H pylori strains and most strains have the s1a genotype, associated with increased virulence. While the high frequency of s1a/m1 vacA genotype might play a role in the increased incidence of atrophic gastritis and gastric cancer in Japanese subjects, it precludes its use as a predictor of clinical outcome of H pylori infection in Japan.
Experience of changing between signal and Bactec 9240 blood culture systems in a children's hospital.
Authors: Gray J , Brockwell M , Das I
Published: January 2009

AIM: To compare experience of positive blood cultures in successive years before and after changing from Signal (Unipath) to Bactec 9240 (Becton Dickinson) blood culture systems. METHODS: Analysis of data collected prospectively on 7967 Signal and 7062 Bactec blood culture sets. RESULTS: Significant growths occurred in 5.7% of Signal and 8.9% of Bactec cultures; 33.0% more significant isolates and 24.0% more episodes of bacteraemia were detected in the second year, following introduction of the Bactec system. Inpatient hospital activity increased by 8.2% between the first and second years, although the numbers of blood cultures received actually fell by 11.4%. There were striking increases in numbers of isolates of coagulase negative staphylococci (47.7%) and Enterobacteriaceae (56.8%) from Bactec cultures. Two anaerobic bacteraemias were detected in Signal blood cultures, whereas none was detected by the Bactec system, despite 12.1% of sets including an anaerobic bottle. Of significant positive cultures, 90.2% were detected within one day with the Bactec 9240, compared with only 50.0% of Signal cultures; 20.7% of significant positive Signal blood cultures were detected only on terminal subculture. Microorganisms that were not significant were isolated from 5.1% Signal and 3.8% Bactec cultures. CONCLUSIONS: Compared with the Signal system, the Bactec 9240 offers markedly more rapid and sensitive detection of bacteraemia, together with a lower rate of non-significant isolates. However, using a single PEDS PLUS/F bottle the few episodes of anaerobic bacteraemia that occur in children are likely to be missed.
The inhibitory effect of phosphate on the ligase chain reaction used for detecting Chlamydia trachomatis.
Authors: Notomi T , Ikeda Y , Okadome A , Nagayama A
Published: January 2009

AIMS: To examine the detection limit of the ligase chain reaction kit for Chlamydia trachomatis, to study the inhibitory effect of phosphate on the ligase chain reaction, and to clarify the mechanism of inhibition. METHODS: Three reference serovars of C trachomatis--D/UW-3/Cx, F/UW-6/Cx, and L2/434/Bu--were used to test the sensitivity of the chlamydia ligase chain reaction. Comparison was made of the inhibition by phosphate before and after DNA amplification. Phosphate in up to 2.4 mM concentration was added to specimens of C trachomatis serovar D (1 to 50 inclusion forming units (IFU)/reaction) before DNA amplification to examine the concentration dependency of phosphate inhibition of the ligase chain reaction. RESULTS: The detection limits were 0.6 IFU/reaction for serovar D/UW-3/Cx and F/UW-6/Cx, and 0.4 IFU/reaction for L2/434/Bu. Phosphate inhibited the ligase chain reaction only when it was added before the amplification stage. The specimens containing chlamydia at 1 to 50 IFU/reaction were negative when the concentration of phosphate added at the prethermocycle stage was more than 1.2 mM. CONCLUSIONS: Ligase chain reaction analysis is a reliable method of diagnosing C trachomatis infection because of its high sensitivity. It would be clearly superior to the currently used methods if the problem of inhibitors could be eliminated. The mechanism of inhibition of the ligase chain reaction by phosphate was thought to be blockade of the amplification of the target DNA. The efficacy of the ligase chain reaction could be inhibited by phosphate in the urine, so duplicate dilution analysis of some negative specimens should be useful.
Comparison of throat swabs with sputum specimens for the detection of Chlamydia pneumoniae antigen by direct immunofluorescence.
Authors: Garnett P , Brogan O , Lafong C , Fox C
Published: January 2009

AIM: To compare throat swabs with sputum specimens for Chlamydia pneumoniae antigen detection. METHODS: During a one year period, sputum and throat swabs from 50 patients over 15 years of age with acute or persisting lower respiratory tract infection were examined for C pneumoniae antigen by direct immunofluorescence. RESULTS: C pneumoniae antigen was detected in 18/50 patients (36.0%) from sputum, throat swab, or both. Paired sputum and throat swabs were received from 35/50 patients (70.0%). C pneumoniae antigen was detected in either or both specimens from 14/35 patients (40.0%). Of the 14 positive patients, both specimens were positive in nine (64.3%), throat swab only in four (28.6%), and sputum only in one (7.1%). Of the remaining 15 patients from whom only a single specimen was sent, a further three of eight throat swabs and one of seven sputum specimens were positive. There was no statistically significant difference between the results obtained from the two types of specimen. CONCLUSIONS: Throat swabs may be as good as sputum for the detection of C pneumoniae antigen.
Do IgA, IgE, and IgG avidity tests have any value in the diagnosis of toxoplasma infection in pregnancy?
Authors: Ashburn D , Joss A W , Pennington T H , Ho-Yen D O
Published: January 2009

AIM: To determine the value of tests for specific IgA, IgE, and IgG avidity in diagnosing Toxoplasma gondii infection during pregnancy. METHODS: In a retrospective study, current serological tests (dye test and three IgM assays with different sensitivities) were compared with immunosorbent agglutination assays (ISAGA) for specific IgA and IgE and an IgG avidity enzyme linked immunosorbent assay (ELISA). Patient group 1 comprised six women with definite or probable infection during pregnancy determined by congenital toxoplasmosis or laboratory results. Group 2 comprised seven women infected during or before 11 pregnancies (two consecutive pregnancies in two patients and three in a third). RESULTS: One patient in group 1 seroconverted during pregnancy. IgA ISAGA and avidity confirmed acute infection when confirmatory IgM ELISA remained negative. In five of six patients from group 1, IgA and IgE ISAGA and avidity confirmed acute infection. In group 2, the dye test titre was raised in seven of 11 pregnancies (six of seven patients). Specific IgM and IgA were positive during all 11 pregnancies. IgE ISAGA was positive in only four of 11 pregnancies (three of seven patients), but negative results in the remainder may exclude acute infection. High avidity antibodies indicative of past infection were found in four of 11 pregnancies (two of seven patients). CONCLUSIONS: Each test improved diagnosis or timing of infection but no single test was ideal. The IgA ISAGA was sensitive and detected seroconversion. Positive IgE ISAGA and low avidity both confirmed infection, whereas negative IgE may exclude acute infection. High avidity diagnosed past infection but persistence of low avidity reduced its value to differentiate acute and past infection. Further studies with larger patient groups are needed to determine the optimum diagnostic strategy. These techniques are valuable in complementing existing tests.
The Vitek analyser for routine bacterial identification and susceptibility testing: protocols, problems, and pitfalls.
Authors: Shetty N , Hill G , Ridgway G L
Published: January 2009

Automated and semiautomated technology in microbiology has seen great advances in recent years. The choice of automated equipment for the identification and susceptibility testing of bacteria in a routine diagnostic laboratory depends on speed, accuracy, ease of use, and cost factors. The Vitek analyser (bioMerieux, UK) was installed in a busy diagnostic teaching hospital laboratory in London. This report describes one year's experience. Changes to work practice as a result of incorporating the equipment into the laboratory, and the advantages and disadvantages of automation in key areas are described in detail, together with possible solutions to problems. The Vitek analyser was found to be valuable for the speed and accuracy with which results were available for the common bacterial pathogens. Results of susceptibility testing were standardised according to NCCLS guidelines and used breakpoint MICs to ascertain susceptibility and resistance; they were an improvement on disc testing. This equipment is not a reference facility for difficult to identify organisms and many manual techniques, including some disc susceptibility testing, will have to be retained by the laboratory.
Apolipoprotein E alleles in women with severe pre-eclampsia.
Authors: Nagy B , Rigó J , Fintor L , Karádi I , Tóth T
Published: January 2009

This study investigated the frequency of apolipoprotein E (apoE) alleles among women with severe pre-eclampsia. The presence of the three most common apoE alleles (epsilon 2, epsilon 3, epsilon 4) was determined by polymerase chain reaction-restriction fragment length polymorphism in three groups of white women: non-pregnant healthy (n = 101), pregnant healthy (n = 52), and pregnant with a diagnosis of severe pre-eclampsia (n = 54). The frequency of apo epsilon 2 was highest among women with severe pre-eclampsia (16.6%) followed by non-pregnant women (12.9%), and those experiencing a healthy pregnancy (10.6%). The higher frequency of the apo epsilon 2 allele detected among women with severe pre-eclampsia suggests that apoE may play a role in the development of pre-eclampsia.
The effect on patient management of temporary non-availability of immunofluorescence for renal biopsy reporting.
Authors: Date A , Dakshinamurthy D S , Jacob C K , Shastry J C
Published: January 2009

Delay in reporting the immunofluorescence findings on renal biopsies, owing to an interruption in supply of reagents, made possible a retrospective analysis of the effect of the lack of this information on patient management. Hospital case records of the 39 patients so affected were reviewed to determine what changes in their management took place after the immunofluorescence findings became available. The clinical, laboratory, and light microscopic findings in all except a case of pauci-immune crescentic glomerulonephritis allowed management decisions to be made that were not influenced by immunofluorescence findings. This was owing to correct prediction of the immunofluorescence findings, as in cases of IgA nephropathy presenting with recurrent haematuria; the adequacy of light microscopy in the interpretation of graft biopsies, in classifying lupus nephritis and in most cases of nephrotic syndrome; and the absence of entities identifiable only by immunofluorescence among these patients.
Obstructive uropathy associated with myelomonocytic infiltration of the prostate.
Authors: Hope-Gill B , Goepel J R , Collin R C
Published: January 2009

A 72 year old man was diagnosed with chronic myelomonocytic leukaemia (CMML) according to the FAB group classification. He presented with symptoms of anaemia, urinary frequency, hesitancy, and nocturia. He was later admitted with acute urinary retention and acute renal failure, which resolved with treatment. A transurethral resection of the prostate was performed. Histological examination showed fibromuscular hyperplasia with dense infiltration by myelomonocytes which stained positively with chloroacetate esterase; immunohistochemical staining was positive for lysozyme, CD43, CD45, and CD68. Following treatment with oral etoposide he transformed to acute myeloid leukaemia and eventually died. Myelomonocytic infiltration of the prostate has not been reported before. This case extends the spectrum of disease previously recognised in CMML.
Screening INR deviation of local prothrombin time systems.
Authors: Poller L
Published: January 2009

AIM: To assess the reliability of local international normalised ratios (INR) using a set of three international reference preparation (IRP) certified freeze dried plasmas. METHODS: 55 centres in the United Kingdom and the Republic of Ireland participated; 36 centres employed coagulometers and 19 a manual prothrombin time technique, all with the same batch of routine commercial thromboplastin. The plasmas had certified INR with the manual technique using a thromboplastin IRP, and results were provided graphically to participants for self assessment. An INR deviation of more than +/- 10% from the certified INR with any of the screening plasmas was regarded as unsatisfactory and clinically significant. Sets of 20 freeze dried plasmas were provided for local ISI calibrations and sets of seven freeze dried normals were provided for supplementary exercises where screening results were unsatisfactory. RESULTS: 15 of 38 coagulometers, but only three of the 19 manual prothrombin time test techniques, gave unsatisfactory results. With 10 of the 15 unsatisfactory coagulometer results the problem was resolved by local ISI calibrations with plasma calibrant sets provided. Unsatisfactory results with manual technique in all four instances were corrected by substitution of the mean result with freeze dried normal plasmas provided. CONCLUSIONS: The freeze dried plasma screening set was useful in detecting incorrect INR in a high proportion of coagulometer users and a smaller number of participants using the manual technique.
Frequency of cervical intraepithelial neoplasia following large loop excision of the transformation zone.
Authors: Hulman G , Pickles C J , Gie C A , Dowling F M , Stocks P J , Dixon R
Published: January 2009

AIM: To determine the frequency of cervical intraepithelial neoplasia (CIN) following large loop excision of the transformation zone of the cervix (LLETZ) according to grade and completeness of excision of CIN. METHODS: A retrospective study of 669 patients who had LLETZ biopsies showing CIN 1, 2, or 3. The patients were subdivided according to the grade and completeness of excision of CIN. The follow up period was 1.5 to 3.5 years. Risk of persistent/recurrent CIN was assessed by the frequency of histological diagnosis of CIN during the follow up period. RESULTS: Frequency of persistent/recurrent CIN increased with the grade of CIN reported: 6.7% of patients with CIN 1, 13.4% with CIN 2, and 21.7% with CIN 3 developed persistence or recurrence. The frequency of CIN persistence/recurrence was significantly lower where LLETZ showed complete excision of CIN (8.4%) than where it was incomplete (31.3%) (p < 0.0001) or equivocal (27.8%) (p < 0.0001). CONCLUSIONS: Patients with incomplete or equivocal excision of all grades of CIN merit careful, preferably colposcopic, follow up. Patients with completely excised high grade CIN require careful cervical cytologic surveillance.
Audit of the histopathological diagnosis of non-neoplastic colorectal biopsies: achievable standards for the diagnosis of inflammatory bowel disease.
Authors: Dube A K , Cross S S , Lobo A J
Published: January 2009

AIM: To assess the performance of a histopathology department in diagnosing inflammatory bowel diseases with comparison of reports from other centres. STUDY POPULATION: 1067 sets of endoscopic biopsies received in the department of histopathology, Royal Hallamshire Hospital, 1990-1995. METHODS: The histopathological diagnosis of non-neoplastic endoscopic colorectal biopsies was audited using data from histopathology reports. The biopsy diagnosis by the initial reporting pathologist and final diagnosis after additional investigations (endoscopy, radiology, microbiology) or surgery were used to derive sensitivity, specificity, and positive predictive values for categories of disease. RESULTS: Diagnosis was validated for 1067 biopsy sets (43% of those initially assessed). For all biopsies (with or without active inflammation) reports highly suggestive or suggestive of Crohn's disease had a sensitivity of 50%; for ulcerative colitis the comparable figure was 62%. Sensitivity was the same for both diagnoses (74%) in those biopsies with active inflammation. Positive predictive values for highly suggestive diagnoses of ulcerative colitis or Crohn's disease were 100%. In all biopsies the specificity of a histopathological diagnosis of normality was 96%. CONCLUSIONS: These results compare favourably with the other published audits and present an achievable level of performance for non-specialist hospitals with non-specialist histopathology services.