Immunofluorescence of frozen tissue sections (IF-F) is a traditional technique used in renal biopsy. However, IF-F has certain disadvantages, such as a few or even no glomeruli in the section, and limited long-term preservation of the fluorescently labelled samples.
We compared two-step immunohistochemistry (IHC) staining of deparaffinised sections for antigen retrieval with microwave combined high-pressure cooking to IF-F used to detect antigens of IgG, IgA, IgM, C3, C1q, and in patient renal biopsy samples. The number of glomeruli detected, sensitivity and specificity of positive staining, tissue structure, and location staining of the antigens were determined using the two methods in 285 patients diagnosed with different renal diseases.
Concordant observations between IF-F and IHC were 99% for all antigen staining (1969 of 1995 observations) and 100% for IgG, IgA and IgM (all 285 observations). The number of glomeruli in IHC sections was significantly greater compared with IF-F sections (p<0.001). IHC provided clearer images of tissue structure, more precise localisation of positive-staining antigens, and IHC staining allowed simultaneous evaluation of tissue by light microscopy. Correlation between tissue structure and immune deposits are not readily attained by IF-F.
IHC is superior to IF-F for immunopathological diagnosis of renal biopsy tissue and is a reliable replacement for the more traditional IF-F method.
To conduct a methods correlation study of three different assays for the detection of mutations at EGFR gene in human formalin-fixed paraffin-embedded tumour (FFPET) specimens of non-small cell lung carcinomas (NSCLC).
We conducted a 2-site method comparison study of two european conformity (CE) in vitro diagnostic (IVD)-marked assays, the cobas EGFR Mutation Test and the Therascreen EGFR29 Mutation Kit, and 2x bidirectional Sanger sequencing. We blind-tested 124 NSCLC FFPET specimens with all three methods; the cobas test was performed at both sites. Positive (PPA) and negative percent agreements (NPA) were determined for the cobas test versus each of the other two methods. Specimens yielding discordant test results between methods were further tested using quantitative massively parallel pyrosequencing (MPP).
PPA between cobas and Sanger was 98.8%; NPA was 79.3%. Overall there were seven discordant results. MPP confirmed an exon 19 deletion in two cases and L858R mutation in four cases. PPA between cobas and Therascreen was 98.9% and NPA was 100%. There was one discordant result. Reproducibility of the cobas test between the two sites was 99.2%.
The invalid rates for the cobas test and Therascreen were lower than Sanger sequencing. The cobas and Therascreen assays showed a high degree of concordance, and both were more sensitive for the detection of exon 19 deletion and L858R mutations than Sanger. The cobas test was highly reproducible between the two testing sites, used the least amount of DNA input and was the only test with automated results reporting.
Biliary neuroendocrine tumours (NETs) are rare and mostly exist as a component of mixed adenoneuroendocrine carcinomas (MANECs). Although the NET component in biliary MANECs is generally more malignant and clinically more important to the prognosis than the ordinary adenocarcinomatous component, the histogenesis of biliary NET has not been clarified. In this study, the role of the Notch1-Hes1 signalling axis in the histogenesis of biliary NETs was examined.
Immunohistochemistry for Notch1, its ligand Jagged1 and Hes1 was performed using surgical specimens from 11 patients with biliary MANEC. Moreover, after the knock-down of Notch1 mRNA expression in a cholangiocarcinoma cell line, the expression of chromogranin A (a neuroendocrine marker) and Ascl1 (a neuroendocrine-inducing molecule inhibited by activated Hes1) was examined by quantitative PCR.
Histological examination revealed that the adenocarcinomatous components were predominately located at the luminal surface of the MANEC and the majority of stromal invasion involved NET components. Ordinary adenocarcinomas and non-neoplastic biliary epithelium constantly expressed Notch1, Jagged1 and Hes1, but the expression of Notch1 and Hes1 was decreased or absent in NET components, suggesting interference with the Notch1-Hes1 signalling axis in biliary NET. Moreover, in the cholangiocarcinoma cell line in which the expression of Notch1 mRNA was knocked down, the mRNA expression of Ascl1 and chromogranin A was increased.
The Notch1-Hes1 signalling axis suppresses neuroendocrine differentiation and maintains tubular/acinar features in adenocarcinoma and non-neoplastic epithelium in the biliary tree. Moreover, a disruption of this signalling axis may be associated with the tumourigenesis of NETs in biliary MANEC.
Complications after liver transplantation are major factors that determine the prognosis of patients. In this study, we aimed at investigating an important though less frequently occurring complication, post-transplant lymphoproliferative disorders (PTLD), in a single institution after liver transplantation.
15 cases with a diagnosis of PTLD in post-liver transplant patients were retrieved from our archive and the clinicopathological features reviewed.
The overall incidence of PTLD was 2.3% (n=15/658), and the incidence was much higher in the paediatric than the adult age groups, being 11.1% (9/81) and 1% (6/577), respectively. The median time of presentation was 16 months after transplantation (occurrence time ranging from 2 to 87 months after transplantation). Lymph nodes, gastrointestinal tract and graft liver were the commonest sites of involvement. 11 cases were classified as monomorphic PTLD according to WHO classification and the majority (n=10/11) of them were of B cell differentiation. 12 of the total 15 PTLD cases showed a positive result for Epstein-Barr virus-encoded RNAs with in situ hybridisation. Eight patients were alive at the time of review, and two of them suffered from recurrence of the PTLD. Among the seven patients who died, six succumbed within 1 year from the diagnosis of PTLD.
Despite its relative rarity as a complication for liver transplantation, PTLD imposes significant effects on the morbidity, mortality and treatment implications in postliver transplant patients. The clinicopathological data would hopefully provide better insight into the surveillance and management for susceptible patients.
of this study is to identify clinical, endoscopic and/or histological features associated with response to treatment with swallowed fluticasone propionate.
In the last 12 years 34 children (M/F 25/9) with EO were treated with fluticasone propionate spray 250 μg/puff by inhaler without spacer, three puffs three times a day for 6 weeks, and returned for a follow-up endoscopy. At histology 25 of them were found to be responders to therapy (73.5%) and 9 were non-responders. Anthropometric characteristics, symptoms at presentation, endoscopic and histological data at baseline between responders and non-responders were compared.
Age, sex, height, duration and type of main symptom at presentation, type of allergy and number of allergens, peripheral eosinophil counts an serum IgE were similar in responders and non-responders. At baseline histology findings responders had a more severe inflammation: median peak eosinophils/high power field was higher (76 vs 44 in non responders p=0.04), eosinophilic microabscesses were present in a significantly higher number of responders (p=0.04) and peak mast cells/ high power field was significantly higher (p=0.001).
Clinical characteristics of children with EO at baseline were similar in responders and non-responders, but a more severe inflammation in oesophageal mucosa was associated with a higher response rate to fluticasone treatment.
Guaiac faecal occult blood tests are being replaced by faecal immunochemical tests (FIT). We investigated whether faecal haemoglobin concentration (f-Hb) was related to stage in progression of colorectal neoplasia, studying cancer and adenoma characteristics in an evaluation of quantitative FIT as a first-line screening test.
We invited 66 225 individuals aged 50–74 years to provide one sample of faeces. f-Hb was measured on samples from 38 720 responders. Colonoscopy findings and pathology data were collected on the 943 with f-Hb≥400 ng Hb/ml (80 µg Hb/g faeces).
Of the 814 participants with outcome data (median age: 63 years, range 50–75, 56.4% male), 39 had cancer, 190 high-risk adenoma (HRA, defined as ≥3 or any ≥10 mm) and 119 low-risk adenoma (LRA). 74.4% of those with cancer had f-Hb>1000 ng Hb/ml compared with 58.4% with HRA, and 44.1% with no pathology. Median f-Hb concentration was higher in those with cancer than those with no (p<0.002) or non-neoplastic (p<0.002) pathology, and those with LRA (p=0.0001). Polyp cancers had lower concentrations than more advanced stage cancers (p<0.04). Higher f-Hb was also found in those with HRA than with LRA (p<0.006), large (>10 mm) compared with small adenoma (p<0.0001), and also an adenoma displaying high-grade dysplasia compared with low-grade dysplasia (p<0.009).
f-Hb is related to severity of colorectal neoplastic disease. This has ramifications for the selection of the appropriate cut-off concentration adopted for bowel screening programmes.
Bone marrow involvement confers a poor prognosis in patients with diffuse, large, B-cell lymphoma (DLBCL). However, the prognostic significance of concordant and discordant bone marrow involvement in these cases differs. We analysed this further in patients treated with R-CHOP (rituximab plus cyclophosphamide, doxorubicin, vincristine and prednisone) at a single institute.
The cytomorphology of bone marrow involvement was evaluated in 632 patients who were diagnosed with DLBCL in primary tissues and had received R-CHOP therapy. Bone marrow trephine biopsies and clot sections were analysed, along with the immunohistochemical analysis of CD20, CD79a and CD3.
Bone marrow involvement was identified in 80 of our DLBCL patient subjects (12.7%). Of these, 32 (40%) showed discordant bone marrow involvement, and 48 (60%) showed concordant involvement. Kaplan–Meier survival analysis showed that progression-free survival and overall survival was poorer in the concordant group (p<0.001). Multivariate analysis, adjusted for the International Prognostic Index score, showed that concordant involvement was an independent predictor of progression-free survival (p<0.001) and overall survival (p=0.011). Discordant involvement was not a negative prognostic factor independent of the International Prognostic Index.
Prognostication based on bone marrow involvement cytomorphology is a useful indicator of progression-free survival and overall survival, independent of the International Prognostic Index score, in DLBCL patients. Accurate staging based on morphology should thus be included in bone marrow examinations of such cases.
Excessively acidic urine is the dominant factor in uric acid stone formation. Recent evidence implicating insulin resistance has revived interest in its causation. We reviewed data on uric acid stone formers attending a general stones clinic to find out whether this supports and adds to current concepts.
A retrospective database study of 1504 stone formers investigated at the Southampton renal stones clinic from 1990 to March 2007. Uric acid stone formers and idiopathic calcium stone formers were compared using non-parametric tests.
Fifty-nine patients (3.9%; 43 men) had uric acid stones. In men the commonest associated conditions were diabetes (20%), gout (20%) and an ileostomy (15%); in women, diabetes (33%), urinary infections (27%) and hyperparathyroidism (20%). Most patients with diabetes (85% of men, 75% of women), however, produced calcium stones. Risk factors did not differ significantly between calcium and uric acid stone formers with diabetes, gout or ileostomies. The median urine pH of men with idiopathic calcium stones was 6.20, idiopathic uric acid stones 5.47, diabetes 5.68, gout 6.05, diabetes and gout 5.20 and ileostomy 5.10. Plasma urate was higher with gout and idiopathic uric acid stones. Urate excretion was increased in gout. Oxalate excretion was lower with idiopathic uric acid stones (new finding). Urine volume decreased and oxalate concentration increased with ileostomy.
Uric acid stones are increased in diabetes, but most patients with diabetes make calcium stones. Different mechanisms may explain low pH with diabetes, gout and idiopathic stones. Low oxalate excretion with idiopathic urate stones needs confirmation.
The UK National External Quality Assessment Scheme for General Haematology (UK NEQAS (H)) and Sysmex UK, in co-operation with Sysmex Europe and University College London Hospital (UCLH), have investigated mean cell volume (MCV) values generated by analysers in the Sysmex X-Class grouping for UK NEQAS (H) full blood count (FBC) surveys, connected to RPU-2100 diluent production units. The RPU-2100 unit uses a demineralised water supply to dilute concentrated CELLPACK reagent to 25 times its volume and then directly supplies up to four connected XE series analysers. This gives the user the benefits of not having to stop the analysers to change the...]]>
We report a case with a history of occupational asbestos exposure in which malignant pleural mesothelioma (MPM) was suspected clinically and diagnosed postmortem as pleural involvement of extrapulmonary small cell carcinoma (SCC). An 85-year-old man with a 65 pack-year history of smoking was referred to our hospital in June 2011. The patient had been exposed to asbestos in the iron production industry over the course of 30 years, and an irregular thickening of the right pleura was observed on chest CT at a medical check-up. The patient had a history of chronic hepatitis C and had been undergone transurethral resection for urothelial bladder cancer five times since 2006. Chest CT revealed neoplastic thickening of the right pleura, which had grown over 6 months (
We present a case of a malignant tumour of the parotid gland, originally thought to represent metastatic melanoma (of unknown primary), later reclassified as malignant melanoma of soft part/clear cell sarcoma (MMSP/CCS) on the basis of the presence of a EWSR1–ATF1 gene fusion. Further immunophenotypic studies demonstrated vascular differentiation in support of a diagnosis of angiosarcoma. Although extraordinarily rare, angiosarcomas of the parotid gland have been described; while MMSP/CCS of the parotid has not been documented, EWSR1 rearrangements have never been demonstrated in angiosarcoma. This case therefore provides an example of the way in which morphological findings must be integrated to provide the appropriate pathological diagnosis.
A 72-year-old otherwise healthy woman presented with a 6-week history of a rapidly enlarging mass of her right jaw, with associated facial nerve paralysis. A head and neck CT scan showed an enhancing 3.5x2.6x4.0 cm mass with ill-defined margins and a somewhat...]]>
PH Tan, AA Thike, WJ Tan et al. Predicting clinical behaviour of breast phyllodes tumours: a nomogram based on histological criteria and surgical margins. J Clin Pathol 2012;65:69–76. There are errors in Tables 1, 8 and 9 of this paper, the following information has been corrected. In Table 1 under ‘Clinicopathological features', Size (mm) (mean 52, median 40, range 3-250) has been changed to: Size (mm) (mean 52, median 40, range 8-250). In Table 8 under ‘Factor', the second entry of Grade has been deleted. The following text has been removed from the footnote: ‘It should be noted that the nomogram has a score range between 0 and 100, but the total histological score has a more limited range between 5 and 13, accounting for the apparently higher HR.'The table footnote now reads: HR (hazard ratio) refers to recurrence risk relative to reference. hpf, high-power fields; NR,...]]>
Volume 1 is dedicated to non-neoplastic lung entities. There is a superb opening chapter on anatomy, histology, embryology and development that lays the foundation for anyone wishing to embark on a career in lung pathology. The diagrams facilitate the understanding of the subject matter. In fact, the illustrations and tables that supplement the text in this book are excellent.
I liked the compendium of consecutive images in Chapter 4, which allow for ‘wall paper matching’ of images and slides.
Volume 2 is ostensibly dedicated to tumours. Here, histological images are supplemented by relevant cytology. The discussion of every tumour and...]]>