Assessment of β-amyloid (Aβ) plaque load in Alzheimer's disease by MRI would provide an important biomarker to monitor disease progression or treatment response. Alterations in tissue structure caused by the presence of Aβ may cause localised changes that can be detected by quantitative T₁ and T₂ relaxation time measurements averaged over larger areas of tissue than that of individual plaques. We constructed depth profiles of the T₁ and T₂ relaxation times of the cerebral cortex with subjacent white matter and hippocampus in six 5xFAD transgenic and six control mice at 11 months of age. We registered these profiles with corresponding profiles of three immunohistochemical markers: β-amyloid; neuron-specific nuclear protein (NeuN), a marker of neuronal cell load; and myelin basic protein (MBP), a marker of myelin load. We found lower T₁ in the 5xFAD transgenic mice compared to wild type control mice at all depths, with maximum sensitivity for detection at specific layers. T₁ negatively correlated with Aβ staining intensity in the 5xFAD mice which had no changes in NeuN and MBP staining compared to wild type mice. We postulate that these relaxation time changes are due to the presence of β-amyloid in the transgenic mice. It may be clinically feasible to develop a similar layered analysis protocol as a biomarker for Alzheimer's disease in humans.

OBJECTIVES:Cerebral small vessel disease (SVD) is common in aged brains and causes lacunar stroke, diffuse white matter lesions (leukoaraiosis), and vascular cognitive impairment. The pathogenesis is unknown. Endothelial dysfunction is a possible causal factor, and circulating markers of endothelial activation (intercellular adhesion molecule-1, thrombomodulin) and inflammation (interleukin [IL]-6) are elevated in patients with SVD. In this case-control study, we tested whether brain endothelial ICAM1, thrombomodulin, and IL-6 are altered in SVD. METHODS:We examined small penetrating cerebral arteries of pathologically diagnosed SVD cases, aged controls without SVD, young control cases with no brain pathology, and cases with early-onset hereditary SVD (cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy [CADASIL]). All tissues had minimal cerebral amyloid angiopathy or other Alzheimer pathology. RESULTS:Thrombomodulin immunoreactivity was present in all aged SVD, aged control, and CADASIL cases, primarily in small artery endothelium. Thrombomodulin was augmented in aged SVD cases compared with aged controls (p = 0.012) and in vessels with higher sclerotic index (an indicator of SVD severity; p < 0.01). Thrombomodulin was sparse/absent in young controls. Endothelial ICAM1 and IL-6 were rarely seen, and were not related to SVD. CONCLUSIONS:Our data suggest that cerebral endothelial activation in deep penetrating arteries is not associated with SVD. Endothelial thrombomodulin increased with SVD severity, and CADASIL data suggest that this may be a cerebral response to SVD. Elevated thrombomodulin may be a protective agent in SVD. Our data confirm endothelial involvement in SVD.

Aim: AL amyloidosis (ALA) is a form of plasma cell (PC) dyscrasia characterised by deposition of insoluble fibrillar deposits in various organs causing organ damage. This is believed to be the first study to evaluate the expression of CD79a, CD20, CD56, cyclin D1 and epithelial membrane antigen (EMA) in neoplastic PCs of ALA.Methods and Results: The study included 36 well-documented cases of ALA. In all cases presence of amyloid deposits had been documented by Congo Red stain in the bone marrow trephine biopsy (BMTB) and/or in other tissues. BMTBs showed varying degrees of PC infiltration (median 12%). In eight of the 36 cases with associated myeloma, the mean (2xSE) percentage of PCs (PC%) was 34 (18)%, while in the remaining, PC% was 15 (4)%. Expression of CD20, CD79a, CD56, cyclin D1 and EMA was noted in 42%, 86%, 50%, 53% and 83% of cases, respectively. Aberrant antigen expression in the form of CD56 and/or cyclin D1 expression was seen in 79% of cases. Nine of 10 cases with small lymphoid-like PCs were positive for CD20 and all the 10 cases were positive for cyclin D1. On the other hand, among cases without small lymphoid-like morphology, CD20 and cyclin D1 expression was seen in only 6 of 26 and 8 of 26 cases respectively (p = 0.001 and 0.002 respectively). CD20 expression correlated with cyclin D1 expression (p = 0.045). Cytological atypia/pleomorphism was predictive of associated myeloma (p = < 0.001).Conclusion: Marrow involvement by neoplastic PCs in ALA can be identified by their aberrant antigen expression apart from light chain restriction, and rituximab as a possible treatment option may be explored in CD20-positive ALA.

Aims: Multiple myeloma ( MM) guidelines in the UK do not advocate performing bone marrow trephine biopsy (BMTB) during follow-up. In a recent study, it was found that the plasma cell per cent (PC%) in BMTB performed at the time of autologous stem cell transplant strongly correlated with survival. The current study addresses whether BMTB is superior to bone marrow aspiration (BMA) in documenting presence of disease and its volume at follow-up.Methods: The study involved 106 samples. A conventional 500-cell differential count was performed on the BMAs to document the PC%. The PC% on BMTBs had been estimated on CD138 immunostain. Furthermore, BMTBs had also been immunostained for CD56, cyclin D1 and light chains.Results: The mean (2SEM) PC% values in BMAs and BMTBs were 13.1 (2.6)% and 31.8 (5.8)% respectively. Based on BMA, BMTB and serum/urine paraprotein or light chain estimation, on 92 occasions (89%) there was detectable disease. The positive predictive value of both BMA and BMTB was 100%, and the negative predictive values for BMTB and BMA were 57% and 22% respectively. Among 98 secretory MM cases, the BMTB-PC% showed significant correlation with paraprotein levels, whereas BMA-PC% did not.Conclusions: It is strongly recommended that BMTB is performed and adequately investigated with immunohistochemistry during follow-up of MM.

HistopathologyVolume 50, Issue 6 p. 810-812 Total CD34+ cells per 10 HPF in bone marrow trephines of patients with chronic myeloid leukaemia correlates with probability of complete cytogenetic response following imatinib treatment V Elliot, V Elliot Departments of Histopathology and Haematology, Hammersmith Hospital & Imperial College, London, UKSearch for more papers by this authorD Marin, D Marin Departments of Histopathology and Haematology, Hammersmith Hospital & Imperial College, London, UKSearch for more papers by this authorD Horncastle, D Horncastle Departments of Histopathology and Haematology, Hammersmith Hospital & Imperial College, London, UKSearch for more papers by this authorK Elderfield, K Elderfield Departments of Histopathology and Haematology, Hammersmith Hospital & Imperial College, London, UKSearch for more papers by this authorJ Howard, J Howard Departments of Histopathology and Haematology, Hammersmith Hospital & Imperial College, London, UKSearch for more papers by this authorJ F Apperley, J F Apperley Departments of Histopathology and Haematology, Hammersmith Hospital & Imperial College, London, UKSearch for more papers by this authorI A Lampert, I A Lampert Departments of Histopathology and Haematology, Hammersmith Hospital & Imperial College, London, UKSearch for more papers by this authorK N Naresh, K N Naresh Departments of Histopathology and Haematology, Hammersmith Hospital & Imperial College, London, UKSearch for more papers by this author V Elliot, V Elliot Departments of Histopathology and Haematology, Hammersmith Hospital & Imperial College, London, UKSearch for more papers by this authorD Marin, D Marin Departments of Histopathology and Haematology, Hammersmith Hospital & Imperial College, London, UKSearch for more papers by this authorD Horncastle, D Horncastle Departments of Histopathology and Haematology, Hammersmith Hospital & Imperial College, London, UKSearch for more papers by this authorK Elderfield, K Elderfield Departments of Histopathology and Haematology, Hammersmith Hospital & Imperial College, London, UKSearch for more papers by this authorJ Howard, J Howard Departments of Histopathology and Haematology, Hammersmith Hospital & Imperial College, London, UKSearch for more papers by this authorJ F Apperley, J F Apperley Departments of Histopathology and Haematology, Hammersmith Hospital & Imperial College, London, UKSearch for more papers by this authorI A Lampert, I A Lampert Departments of Histopathology and Haematology, Hammersmith Hospital & Imperial College, London, UKSearch for more papers by this authorK N Naresh, K N Naresh Departments of Histopathology and Haematology, Hammersmith Hospital & Imperial College, London, UKSearch for more papers by this author First published: 21 March 2007 https://doi.org/10.1111/j.1365-2559.2007.02648.xRead the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Volume50, Issue6May 2007Pages 810-812 RelatedInformation

AIMS:To validate and improve the existing algorithm (proposed by Hans et al.) to classify diffuse large B-cell lymphoma (DLBCL).METHODS AND RESULTS:Tissue microarrays constructed from 81 patients with DLBCL were studied by immunohistochemistry for expression of CD10, Bcl-6, MUM1, Bcl-2, cyclin-D2, FOXP1 and PKC-gamma proteins. Cases were classified as either germinal centre B-like (GCB) or non-GC according to Hans et al. An alternative classification was also employed, in which cases positive for either CD10 or Bcl-6 were considered as a GC subgroup and cases negative for both CD10 and Bcl-6 were considered as a non-GC subgroup. GC was further subdivided into favourable GC (negative for both Bcl-2 and cyclin-D2) and unfavourable GC (positive for either Bcl-2 or cyclin-D2). The 5-year event-free survival (EFS) amongst patients classified as favourable GC versus 'others' was 49.5% and 7.3%, respectively (log rank P < 0.0001). Similarly, the 5-year overall survival (OS) amongst patients classified as favourable GC versus 'others' was 58.6% and 13.7%, respectively (log rank P = 0.0001). The difference in survival was independent of the international prognostic index.CONCLUSIONS:In this group of patients the risk stratification based on the new algorithm was better than that proposed by Hans et al.

Specimens of bone marrow trephine biopsy (BMT) are transported and fixed in acetic acid–zinc–formalin fixative, decalcified in 10% formic acid–5% formaldehyde and processed with other specimens to paraffin-wax embedding. Sections, 1-μm-thick, are cut by experienced histotechnologists and used for haematoxylin and eosin, Giemsa, reticulin silver and other histological stains. Further, all immunohistochemical procedures used in the laboratory, including double immunostaining, can be used on these sections with no or minimal modifications. About 10 000 BMT specimens have been analysed using this procedure since 1997 and diseases involving the bone marrow have been classified successfully. More recently, standardised polymerase chain reaction-based analysis and mRNA in situ hybridisation studies have been conducted. Excellent morphology with good antigen, DNA and RNA preservation is offered by the Hammersmith Protocol.

We studied the expression of CD34 in 169 bone marrow trephines of 53 patients with CML in chronic, accelerated and blastic phases. The bone marrow blast count ranged from 0–82% (median: 4.5%). All were receiving imatinib treatment. We aimed to establish which counting method in bone marrow trephines provided the best correlation with the blast count of the corresponding bone marrow aspirate in all phases of the disease. Paraffin embedded sections were stained with the monoclonal antibody QBEND10.

We studied expression of CD34 positive cells in the bone marrow trephines of two groups of patients with CML treated with imatinib. In the first group (n=17), patients went on to achieve sustained complete cytogenetic response during their disease course. In the second group (n =33) complete cytogenetic response was not achieved and patients later developed either accelerated or blastic phase disease. We studied paraffin embedded bone marrow trephines stained with the monoclonal antibody QBEND10 during an eighteen month period following initiation of imatinib therapy and recorded the highest CD34 positive cell counts and aspirate blast counts in three six-month intervals. CD34 positive cells were evaluated using three counting methods. The first method, established in our department for several years, expressed the percentage of CD34 positive cells vs. all nucleated cells in a minimum 500-cell count (% CD34+/500 cells). The second recorded the total number of CD34 positive cells in ten high power fields (CD34+/10hpf) using x 40 magnification /numerical aperture 0.65 and the third method utilised the highest number of CD34 positive cells expressed in a single high power field (CD34+/hpf), the value being taken from the previous CD34+/10 hpf count. We hypothesised that the overall CD34 positive cell burden and absolute CD34 positive numbers would be more important in predicting the likelihood of complete cytogenetic response and this would be better assessed by volumetric evaluation than percentage calculation relative to other nucleated cells. CD34 expression in endothelial cells was taken as a positive internal control. We aimed to establish which variable: % blast count, % CD34+/500 cells, CD34+/10hpf and CD34+/hpf provided better correlation with subsequent cytogenetic response to treatment. Values for each variable were dichotomised at the following levels; aspirate blast count 5, % CD34+/500 cells 2%, CD34+/10hpf 60 cells and CD34+/hpf 10 cells. Univariate analysis was performed on samples within the first six months, seven to twelve months and thirteen to eighteen months. Only CD34+/10hpf showed significant discriminatory ability throughout the entire eighteen month period (Pearson’s Chi-square 2-sided sig: 0.031, 0.003 and 0.007 in the first six months, seven to twelve months and thirteen to eighteen months samples respectively). By logistic regression, we confirmed that CD34+/10hpf had the best correlation with cytogenetic response during the first year following initiation of imatinib therapy. Larger studies are required to confirm these initial findings and to establish volumetric assessment of CD34 positive cells as a predictor for cytogenetic response.