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Bone marrow biopsy depends on tissue morphology, immunohistochemical staining, and moleculardetection. Tissue pretreatment is required for bone marrow samples, from clinical specimen acquisition to pathological reporting, but during the process, proteins and nucleic acids are often altered because of the acid in fixation and decalcification solutions. In our study, we present an easy and effective pretreatment protocol and compared this novel pretreatment protocol (Set 2) with an existing traditional pretreatment process (Set 1) using tissue morphology, IHC staining, and molecular pathological analyses. Granulocytic IHC markers showed more intensive staining in samples of Set 2 than in those of Set 1. The Set 2 protocol provided a higher DNA yield and less fragmentation; moreover, samples processed with the Set 2 protocol could be subsequently used in FISH and DNA sequencing assays. Our optimized novel pretreatment protocol could better protect proteins and DNA molecules while maintaining good cell morphology compared to traditional pretreatment The novel pretreatment reagents could role as a reference by more laboratories for pretreating bone marrow biopsy samples and scientific research. [ABSTRACT FROM AUTHOR]

We evaluated the impact of bone marrow sample characteristics on the detection of persistent cytogenetic abnormalities (PCA) following induction chemotherapy for acute myeloid leukemia (AML). PCA’s were identified in 20.4% of patients and were more common with complete remission without count recovery (CRi) vs. those with count recovery (CR, 45.8 vs. 13.5%,p = .001), with  >2% blasts vs.  ≤2% blasts (42 vs. 12%,p =  .001) and with hypocellular trephine biopsies relative to those with normo/hypercellular biopsies (42.1 vs. 17.3%,p  = .03), although in a multivariate analysis only CRi and blast count >2% were independently associated with a PCA. PCA’s were not observed in patients with favorable risk karyotype. Amongst patients with intermediate and unfavorable risk karyotypes PCA were not associated with differences in overall or, amongst non-transplanted patients, relapse free survival. Thus, although PCAs are common post-induction it is unclear whether they provide any independent prognostic information beyond the diagnostic karyotype. [ABSTRACT FROM AUTHOR]

Objective: To determine the histological outcome of pancytopenia cases on bone marrow trephine biopsy and to see the frequency of various causes of pancytopenia in our population. Study Design: Descriptive study. Place and Duration of Study: Pathology department, Combined Military Hospital (CMH), Kharian (Pakistan). One year (Jan 2015-Dec 2015). Material and Methods: Two hundred bone marrow trephine biopsies were done in one year (2015), out of which 40 were done for evaluation of pancytopenia. The criteria for diagnosis of pancytopenia were; haemoglobin less than 10 g/dl, total leukocyte count (TLC) less than 4.0 x 109/l and platelet count less than 100,000 x 109/l. Patients with pancytopenia secondary to drugs, chemotherapy and radiotherapy were excluded from the study. Trephine biopsies showing marked crushing and having inadequate material were also excluded from the study. Biopsies were processed, slides made and examined under light microscope by haematologist and histopathologist. Frequencies of various causes of pancytopenia diagnosed on histopathology were calculated. The findings were analyzed by using SPSS version 10.0. Result: Out of 40 cases of pancytopenia, male to female ratio was 3:2. The age range was between 1 year to 75 years. Histopathological analysis of bone marrow trephine biopsies revealed megaloblastic anaemia as the most common cause of pancytopenia (30%), followed by aplastic anaemia (25%) and hypersplenism (15%). Conclusion: Megaloblastic anaemia is the most common cause of pancytopenia in our population as compared to aplastic anaemia mentioned in most of the international studies. This indicates prevalence of nutritional deficiency in our population and megaloblastic anaemia must be kept at top of list while evaluating pancytopenia cases. Early diagnosis and treatment of megaloblastic anaemia will prevent any further complication of this disease. [ABSTRACT FROM AUTHOR]

Most guidelines suggest that only the bone marrow aspirate (BMA) is necessary to assess residual disease following intensive chemotherapy for Acute Myeloid Leukemia (AML) with the bone marrow trephine biopsy (BMTB) recommended in cases of a poor quality BMA. We performed a retrospective study evaluating this in a cohort of patients receiving intensive chemotherapy for AML. Residual disease was assessed by morphological examination of the BMA and BMTB ± immunohistochemistry. Of the 647 marrows 32.6% were interim marrows performed prior to peripheral count recovery, 41.7% were end of induction (EOI) marrows and the remaining were ‘other marrows’. The BMA and BMTB findings were concordant in 92.8% of cases. The BMTB led to a change in diagnosis from ‘no leukemia’ to ‘residual leukemia’ in 5.2% of interim, 3.7% of EOI and 2.4% of ‘other’ marrows. The BMA alone had a sensitivity of 86.8% in detecting residual leukemia and of 82.3%, 82.5% and 94.2% for interim, EOI and ‘other marrows’, respectively. Despite the high concordance between the BMA and the BMTB the poor sensitivity of the BMA in detecting residual leukemia, particularly at EOI, may lead to an overestimation of the complete remission rates which may have therapeutic and clinical trial implications. [ABSTRACT FROM AUTHOR]

This study aimed to document the morphological and immunophenotypic features, and describe the diagnostic features of bone marrow ( BM) involvement in human herpes virus 8 Multicentric Castleman disease ( HHV8- MCD). BM trephine biopsy ( BMTB) specimens from 28 patients were revisited. Samples were evaluated for expression of CD3, CD20, CD138, CD68R, glycophorin C, CD42b, HHV8-latency-associated nuclear antigen ( LANA1), Epstein-Barr virus-encoded small RNA and light chains. Presence of significant numbers of HHV8- LANA1+ lymphoid/plasmacytic cells, noted in 10/28 cases, was indicative of BM involvement and was associated with low CD4 and CD8 counts in peripheral blood. The characteristic morphological appearance of MCD seen in lymph nodes is a rare finding in BMTB. 4/5 cases with lymphoid aggregates were involved by MCD, whereas 6/23 cases without lymphoid aggregates were involved by MCD ( P = 0·023). 9/18 cases with hypercellular marrow were involved by MCD, whilst only 1/8 cases with normo/hypocellular marrow showed involvement by MCD ( P = 0·070). While 9/21 cases with increased marrow reticulin were involved by MCD, none of the cases with no increase in reticulin were involved by MCD ( P = 0·080). Reactive plasmacytosis is a frequent finding. We conclude that bone marrow is involved in a significant proportion of patients with MCD (36%), and involvement can be identified by HHV8- LANA1 immunohistochemistry. [ABSTRACT FROM AUTHOR]

Myelodysplastic syndrome (MDS) represents a heterogeneous group of clonal disorders of haematopoietic bone marrow (BM) stem cells manifesting as ineffective haematopoiesis and morphologically visible dysplasias, leading to peripheral cytopenias involving one or more blood cell lineages. The clinical course of the disease is variable and individual. It may lead either to the onset of fatal cytopenias due to progressive BM failure or, on the contrary, to the development of secondary acute myeloid leukaemia (AML). Diagnosis of the disease requires close correlation of the clinical picture, laboratory parameters and standardized evaluation of BM biopsy. Despite some progress in the treatment of patients with higher risk MDS (IPSS group int-2 and high), the current therapeutic options are limited and usually allogeneic haematopoietic stem cell transplantation, chemotherapy, supportive care and hypomethylating therapy may be considered. From 2008-2014, 76 patients were identified with high-risk MDS in our biopsy register who were treated with chemotherapy, the hypomethylating agent azacytidine or stem cell transplantation and whose bone marrow biopsies were available before and after treatment. The effect of the applied therapy was evaluated with focusing on the histopathological type of MDS before and after therapy and also on any change of the histo-morphological picture, particularly the BM blast count, progression to secondary AML and progression of myelofibrosis. The series consisted of 76 patients (47 men and 29 women); the average age was 62 years (27-82 years). The most common type of high-risk MDS was RAEB 2 (32 patients) and secondary AML (21 patients). 39 patients were treated only with the hypomethylating agent azacytidine; in 12 cases chemotherapy was also administered. 22 patients were treated with chemotherapy alone. In 3 patients, only information about peripheral stem cell transplantation was available, the total number of transplantations was 7. Hypomethylating therapy resulted in: a) blast cell count reduction in 16% of cases, b) disease stabilization without striking changes of the morphological picture in nearly 42% of cases, c) rapid disease progression despite therapy in approx. 35% of cases and d) gradual progression of myelofibrosis during the disease course in 8% of cases. Whether the progression of fibrosis represents an effect of hypomethylating treatment or is part of the natural course of MDS remains unknown, the same is true for the persistence or progression of cytopenia/s. [ABSTRACT FROM AUTHOR]

Objectives Bone marrow (BM) aspiration and trephine biopsy is one of the most valuable procedures in the evaluation of hematological disorders. There is a shortage of published literature regarding the indications, procedure, and outcome of bone marrow examination (BME) in neonates and infants. The aim of the present study is to analyze the common indications of performing BME and to assess the spectrum of disorders diagnosed from BM of neonates and infants. Methods A retrospective analysis of BMEs performed in infants over a period of 5 years, between 2009 and 2013 was done. Results and discussion A total of 297 BME were performed on 285 infants, which constitutes 10.3% of pediatric BME procedures during the same period. In our institute, BME is routinely performed by trained pathologists from posterior superior iliac spine in children including infants and neonates with an overall sample adequacy of 97%. Evaluation of cytopenias and suspicion of storage disorder were the most common indications for BME procedure, while acute leukemias and storage disorders were the most common diagnoses offered in infant BM. Conclusions Posterior superior iliac spine is a good site of BME in neonates and infants. BM trephine biopsy is a difficult procedure in this age group, however remains indispensable in situations where an infiltrative pathology is suspected. BME not only helps to make specific diagnoses but should also be used as an extremely valuable, quick, and economically viable procedure to exclude major hematological disorders including certain forms of storage disorder and hematological malignancy in this age group. [ABSTRACT FROM AUTHOR]

Background: The bone marrow is frequently involved in variety of cases presenting with hematological and non-hematological disorders, which are diagnosed by two separate but interrelated techniques such as bone marrow aspiration (BMA) and bone marrow biopsy (BMB). Aim: This study was aimed to assess the diagnostic value of the BMA and BMB and role of both the procedures to reach final diagnosis when done simultaneously. Settings and Design: It was a prospective study. The findings of BMA smears were correlated with BMB sections and data obtained was analysed. Materials and Methods: BMA and BMB were performed on 50 patients. Criteria of inclusion included the main indications for performing this procedure, the availability of full medical records and patient consent. The patients had a male to female sex ratio of 1.6:1 and a wide age range from 4 years to 74 years. Results: In the present study, the main indications for bone marrow examination were categorized. Out of 50 cases studied, in 23 cases, a strong positive correlation between BMA and BMB was noted. However, it was found that in the cases of aplastic anaemia, different phases of myeloproliferative neoplasm (MPN), multiple myeloma, tubercular granulomas and hemato-lymphoid neoplasm, involvement of the marrow was detected better in bone marrow biopsies. Conclusion: The study concludes that preparations of aspirate and trephine biopsy are easy, rapid and complementary to each other in majority of the lesions. The advantage of both the procedures done together enabled us to study the cytomorphology of the cells along with the pattern of distribution of the cells depending on the cases, hence help in making the diagnosis accurately. [ABSTRACT FROM AUTHOR]

Abstract: Evaluation of remission in adult acute lymphoblastic leukemia (ALL) normally relies on cytologic evaluation and flow-cytometric analysis. The diagnostic impact of bone marrow trephine biopsies has not been studied so far. We investigated 26 biopsies of 16 patients with T-ALL. Double fluorescent immunostaining with TdT and CD3 was performed. Corresponding cytologic evaluation and flow-cytometric data were available. In 17 of 26 investigations, the results were concordant (either positive or negative). In one examination, residual blasts were not recognized by trephine biopsy, but by other methods. By contrast, in eight investigations, the leukemic blasts were found only in the bone marrow trephine biopsies. Regarding the evaluation of remission of T-ALL, in our retrospective study, bone marrow trephine biopsies with double immunostaining were found to be sensitive and specific for the detection of residual blasts. This method may add to the conventional cytology and flow-cytometric analysis. [Copyright &y& Elsevier]

Accurate staging of Hodgkin’s lymphoma (HL) is necessary in selecting appropriate treatment. Bone marrow trephine biopsy (BMB) is the standard procedure for depicting bone marrow involvement. BMB is invasive and explores a limited part of the bone marrow. 18F-FDG PET/CT is now widely used for assessing response to therapy in HL and a baseline study is obtained to improve accuracy. The aim of this retrospective analysis was to assess whether routine BMB remains necessary with concomitant 18F-FDG PET/CT. Data from 83 patients (newly diagnosed HL) were reviewed. All patients had received contrast-enhanced CT, BMB and 18F-FDG PET/CT. Results of BMB were not available at the time of 18F-FDG PET/CT imaging. Seven patients had lymphomatous involvement on BMB. Four patients had bone involvement on conventional CT (two with negative BMB). All patients with bone marrow and/or bone lesions at conventional staging were also diagnosed on 18F-FDG PET/CT scan. PET/CT depicted FDG-avid bone/bone marrow foci in nine additional patients. Four of them had only one or two foci, while the other had multiple foci. However, the iliac crest, site of the BMB, was not involved on 18F-FDG PET/CT. Osteolytic/sclerotic lesions matching FDG-avid foci were visible on the CT part of PET/CT in three patients. MRI ordered in three other patients suggested bone marrow involvement. Interim and/or end-therapy 18F-FDG PET/CT documented response of FDG-avid bone/bone marrow foci to chemotherapy in every patient. 18F-FDG PET/CT highly improves sensitivity for diagnosis of bone/bone marrow lesions in HL compared to conventional staging. [ABSTRACT FROM AUTHOR]

Objectives: To better realize the features of peripheral blood (PB), bone marrow (BM) aspirate and especially BM trephine biopsy in atypical chronic myeloid leukemia (aCML). Methods: We studied PB, BM smears in 35 cases of aCML and compared with 84 cases of chronic granulocytic leukemia chronic phase (CGL-CP), 39 cases of chronic myelomonocytic leukemia (CMML). In addition, we evaluated characteristics of BM trephine biopsies in 21 cases of aCML and compared with 68 cases of CGL-CP, 20 cases of CMML. Results: All aCML patients presented with leukocytosis (median WBC 17.3 × 109/L), 48% had moderate anemia, and 85% had thrombocytopenia. Values of monocytes, eosinophils, basophils, percentage of immature granulocytes and monocytes (0.63 ± 0.41 × 109/L, 0.18 ± 0.16 × 109/L, 0.09 ±0.08 × 109/L, 6.27 ± 3.09%, and 2.46 ± 1.75%, respectively) were useful in distinguishing aCML from CGL-CP and CMML groups. The BM smears showed that striking dysgranulopoieis (100%), dyserythropoiesis (48.6%), percentage of blasts, nucleated erythrocytes, monocytes, eosinophils, and basophils (2.45 ± 2.06%, 7.76 ± 2.89%, 1.30 ± 1.21%, 1.47 ± 1.60%, and 1.15 ± 1.08%, respectively) were all important parameters for a diagnosis of aCML. On BM trephine sections, aCML was characterized as hypercellularity, a moderate degree of reticulin fibrosis (71.4%), lymphocytopenia (76.2%), plasmacytopenia (90.5%), abnormal localization of immature precursors (28.5%), and absence of eosinophilia, basophilia, monocytosis. Furthermore, BM imprints, immunohistochemical, and cytochemical staining findings provided important morphological reference to BM trephine sections and made the identification of nucleated cells more convenient. Conclusions: Besides the findings observed in PB and BM aspirate, features of BM trephine biopsy (including BM trephine section, BM imprint, immunohistochemical, and cytochemical staining) can also aid in the diagnosis of aCML. [ABSTRACT FROM AUTHOR]

Abstract: We identified different phenotypic subsets among 62 cases of chronic myeloid leukemia (CML) in blast crisis (BC) (26% B-lymphoblastic, and 74% various myeloblastic subsets) on bone marrow trephines and correlated the blast-phenotype with cytogenetics. Five of myeloid-BC had an associated 3q26 abnormality and two of these showed a megakaryoblastic-phenotype. While myeloid-BC was associated with additional copies of Philadelphia (Ph) (29%) (p =0.08), numerical abnormalities (51%) (p =0.007), trisomy-8 (29%) (p =0.08) and 17p-loss (22%), none of lymphoid-BC showed these abnormalities. Among myeloid-BC, CD34-negative cases were more often associated with trisomy-8, 17p-loss and numerical abnormalities, and the CD117-negative subset with additional copies of Ph (p <0.05). [Copyright &y& Elsevier]

Abstract: Atypical chronic myeloid leukemia (aCML) as defined by the WHO classification is a rare hematopoietic stem cell disorder, which shows both myeloproliferative as well as myelodysplastic features. Because of the presence of neutrophilic leukocytosis, aCML may resemble chronic myelogenous leukemia. However, in contrast with the latter, aCML lacks a Philadelphia chromosome or the BCR/ABL fusion gene. The molecular pathogenesis of aCML and its relationship to other myeloproliferative neoplasms is unknown. To clarify these points, the presence of JAK2 V617F was examined by a retrospective analysis of archival specimens obtained from two large medical institutions. Paraffin-embedded bone marrow (BM) trephines and clot sections were examined by an allele-specific TaqMan PCR suitable for use with decalcified tissue. Fifty-nine cases of Philadelphia (Ph) chromosome negative chronic myeloproliferative neoplasms (CMPN) and normal bone marrows (BM) served as controls. None of the nine amplifiable cases of aCML and none of the normal BM controls showed a JAK2 V617F mutation, in contrast to 45/59 (76%) of the Ph chromosome negative CMPN cases. Atypical CML should therefore be considered as a JAK2 negative chronic myeloid neoplasm that remains properly categorized, alongside chronic myelomonocytic leukemia and juvenile myelomonocytic leukemia, within the WHO group of myelodysplastic/myeloproliferative neoplasms. [Copyright &y& Elsevier]

Bone marrow (BM) examination is a routine staging procedure in follicular lymphoma (FL). Commonly, both BM histology as well as flow cytometry (FCM) of BM aspirates are performed. In order to compare the diagnostic value of these two techniques, we retrospectively evaluated trephine BM biopsies and listmode data of patients with a confirmed diagnosis of FL, obtained in parallel during a 5-year period. One hundred and thirty nine specimens from 91 patients with FL were eligible for analysis. After joint review, nine cases (6.5%) were reclassified, either in histology (six cases) or FCM (three cases). Seventy nine specimens (57%) showed no infiltration with both methods. Sixty specimens (43%) were scored positive for BM involvement by any of the two techniques. Concordant positive results were obtained in 41 cases (68% of positive BM). False negative results were obtained by FCM in 14 cases (23% of positive BM) and by histology in five cases (8%). Discrepant results between BM histology and FCM are frequent in patients with FL, most likely due to the predominantly paratrabecular infiltration and fibrosis typical for FL. Due to the lower false negative rate, trephine BM biopsy remains crucial for the detection of BM involvement in FL. [ABSTRACT FROM AUTHOR]

Akkaya H, Dogan O, Agan M, Dincol G. The value of tatrate resistant acid phosphatase (TRAP) immunoreactivity in diagnosis of hairy cell leukemia. APMIS 2005;113:162–6.Hairy cell leukemia (HCL) is a rare chronic B-cell lymphoproliferative disorder characterized by splenomegaly, pancytopenia, and circulating atypical lymphocytes with circumferential cytoplasmic projections. We investigated the specificity and the sensitivity of anti-TRAP antibody immunoreactivity in 57 cases of HCL. We found that there is a statistically highly significant difference between TRAP immunoreactivities of the study and the control groups, and HCL can be diagnosed by TRAP immunoreactivity in bone marrow trephine biopsy materials with a specificity of 98.27 % and a sensitivity of 100%. [ABSTRACT FROM AUTHOR]

A survey of morbidity and mortality of bone marrow aspiration and trephine biopsy was carried out for the British Society of Haematology, covering the 12 months of 2002. Fifty-three centres reported 13 506 procedures, comprising 3927 aspiration biopsies and 9579 combined procedures. There were 17 adverse events including nine instances of haemorrhage, four infections and one haemorrhage complicated by infection. Two trephine biopsy needles broke during the procedure. One patient suffered considerable pain for 2 weeks. The adverse event may have contributed to death in two patients and in a third patient was life-threatening. Risk factors for adverse events were identified. [ABSTRACT FROM AUTHOR]

Severe retroperitoneal hemorrhage represents an infrequent and serious complication of bone marrow biopsy. A 53-year-old man, diagnosed with polycythemia vera 12 years earlier, was submitted to a bone marrow biopsy due to the appearance of anemia with clinical and hematological features suggesting myelofibrotic transformation, a diagnosis that was confirmed by the marrow study. At 2 h of a right anterior iliac bone marrow trephine biopsy, the patient suddenly developed severe pain in the area of the biopsy, with antialgic flexion of the right leg. Computed tomographic (CT) scan of the abdomen showed a 5 x 9.5 cm hematoma in the right iliac and psoas muscles. The patient was initially managed with analgesics and transfusional support, but the pain persisted and a continuous fall in the hematocrit was observed in the following days. Angiographic examination of the right external iliac artery showed contrast extravasation arising from the circumflex iliac branch, which was embolized using polivinyl alcohol particles and one coil. Following such procedure, the patient recovered uneventfully and was discharged in good condition a few days later. This case illustrates the effectiveness of an endovascular approach in providing a fast and minimally invasive treatment for this life-threatening complication of bone marrow trephine biopsy. [ABSTRACT FROM AUTHOR]