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Marrow Aspirates (marrow + aspirate)
Kinds of Marrow Aspirates Selected AbstractsA novel mutation in the last exon of ATRX in a patient with , -thalassemia myelodysplastic syndromeEUROPEAN JOURNAL OF HAEMATOLOGY, Issue 5 2006Daniel B. Costa Abstract:, We describe a patient with acquired alpha-thalassemia myelodysplastic syndrome (ATMDS). A previously healthy 66-year-old man presented with hemoglobin of 9.3 g/dL, mean corpuscular volume 59 fL, and a bone marrow aspirate with increased erythroid precursors and hypolobulated megakaryocytes. Hemoglobin H inclusions were seen in most red cells after 1% brilliant cresyl blue supravital stain of the peripheral blood. At the molecular level, we identified of a novel mutation in the most 3, exon of the ATRX gene (CGA,TGA substitution in codon 2407) resulting in a premature termination codon (p.R2407X). This case provides further evidence for a link between ATRX mutations and ATMDS, and suggests a possible role for the conserved Q-box element in ATRX function. [source] Possible automatic cell classification of bone marrow aspirate using the CELL-DYN 4000® automatic blood cell analyzerJOURNAL OF CLINICAL LABORATORY ANALYSIS, Issue 2 2002Ryousuke Yamamura Abstract In clinical hematology, the demand for bone marrow aspiration testing is increasing. However, conventional automatic blood cell analyzers cannot completely analyze erythroblasts, and evaluation has mainly been performed by visual examination (the microscopic method). Using the CELL-DYN 4000® automatic blood cell analyzer (CD4000) (Abbott Laboratories, North Chicago, IL), specific recognition and classification of erythroblasts by DNA staining is possible. In the present study, using bone marrow blood collected from normal subjects and patients with hematological malignancy, we classified cells by the microscopic method and with the CD4000, and compared the results. Good correlations were found for total nucleated cell count (TNCC), neutrophils, lymphocytes, erythroblasts, and the myeloid series to erythroid series (M/E) ratio. It is possible to detect blasts that emerge in patients with hematological malignancy using the blast flag system installed on the CD4000. Since all of the items can be analyzed in about 80 sec with the CD4000, cells in bone marrow aspirates can be classified faster with this apparatus than by the microscopic method. Therefore, analysis of bone marrow aspirates with this apparatus appears to be very useful not only for laboratory testing but also for clinical screening. J. Clin. Lab. Anal. 16:86,90, 2002. © 2002 Wiley-Liss, Inc. [source] Intended management of children with acute idiopathic thrombocytopenic purpura: A national surveyJOURNAL OF PAEDIATRICS AND CHILD HEALTH, Issue 1-2 2005MK Marks Objective: In Australia acute idiopathic thrombocytopenic purpura (ITP) is mainly treated by paediatricians (either general paediatricians or paediatric haematologists/oncologists). A survey was conducted to gauge the current practice of treating children with acute ITP in Australia. Methods: All practising Australian paediatricians registered by the Royal Australasian College of Physicians were surveyed regarding their intended management of children with acute ITP. The questionnaire, adapted from a study of paediatric haematologists/oncologists in North America, presented four clinical scenarios of children with acute ITP with a platelet count of 3000 × 109/L, with and without mucosal bleeding (wet and dry purpura, respectively). Questionnaires were returned by mail or filled in online at a dedicated webpage. Results: Five hundred and sixty-three of 1097 (51%) paediatricians responded to the survey. Data from 140 who had treated at least one child with ITP in the previous 12 months were analysed. Respondents indicated that children with acute ITP are usually or always hospitalised (58,92%) and that 48% would be given active treatment, even with dry purpura. Various regimens of i.v. immunoglobulin or corticosteroids are used when treatment is administered. In comparing Australian and North American management of acute ITP there were many similarities, although Australian paediatricians were less likely to arrange a bone marrow aspirate if corticosteroids were prescribed. Conclusions: There is great variation in the intended management of children with acute ITP in Australia. Previously published management recommendations regarding investigation and treatment have had little impact on intended practice. Prospective studies are required to evaluate hypotheses so as to produce evidence-based recommendations for treatment of patients with acute ITP. [source] Canine leishmaniasis with nephrotic syndrome and aortic and caudal vena cava thromboembolismJOURNAL OF VETERINARY EMERGENCY AND CRITICAL CARE, Issue 5 2008Nuno Félix DVM Abstract Objective , To describe a case of leishmaniasis associated with nephrotic syndrome and aortic and caudal vena cava thrombosis in a dog. Case Summary , A 3-year-old male Boxer was referred to the Faculty of Veterinary Medicine, Lisbon, with vomiting, polyuria, polydipsia, lethargy, anorexia, and weight loss. On admission, the dog was thin, quiet, and dehydrated. Initial laboratory abnormalities were compatible with a diagnosis of leishmaniasis (confirmed by serology and bone marrow aspirate), and nephrotic syndrome. Three days later, the animal developed lumbar pain, paraparesis, and absent femoral pulses. Coagulation tests showed a marked reduction in antithrombin (AT) and a mild increase in serum fibrinogen concentration. A diagnosis of thromboembolism was made. In spite of treatment aimed at controlling the primary condition and decreasing further thrombus formation, necrosis developed in the distal right pelvic limb and the nail beds of the left pelvic limb. Against medical advice, medication was stopped and, 15 days later, the dog returned to the hospital, showing extensive necrosis of both pelvic limb extremities. Euthanasia was performed at the owner's request. Necropsy showed a thrombus localized at the distal aorta and extending into the right iliac artery, and an additional thrombus extending from both femoral veins onto the caudal vena cava. New or Unique Information Provided , Thromboembolic disease is rare in dogs with leishmaniasis with nephrotic syndrome. This case suggests that a marked decrease in AT and a mild increase in serum fibrinogen may elicit a hypercoagulable state in these patients. [source] Oral leishmaniasis in a HIV-positive patient.ORAL DISEASES, Issue 1 2002Report of a case involving the palate Leishmaniasis is a parasitic disease caused by a protozoon (Leishmania), with different clinical forms that are endemic in certain countries. The association of this disease in patients who are seropositive to human immunodeficiency virus (HIV) has recently been described. Leishmaniasis can develop in any stage of HIV infection, although the clinical manifestations , and hence the diagnosis , tend to coincide with the periods of maximum immune depression. We present the case of a HIV-positive, ex-intravenous drug abuser (in stage B2 of the CDC, 1992) with concomitant hepatitis C infection who presented with palatinal pain and bleeding for the past 2 months. Exploration revealed a vegetating tumoration of the hard palate. Hematoxylin-eosin and Giemsa staining of the biopsy confirmed the diagnosis of leishmaniasis. The definitive diagnosis was mucocutaneous leishmaniasis (MCL), for a bone marrow aspirate proved negative, and no further lesions could be established. The patient was treated with meglumine antimoniate (Glucantime), followed by improvement of the lesions. [source] More aggressive bone marrow screening in retinoblastoma patients is not indicated: The memorial Sloan-Kettering cancer center experience,PEDIATRIC BLOOD & CANCER, Issue 1 2006Stergios Zacharoulis MD Abstract Background Bone marrow involvement in retinoblastoma patients is rare in the more industrialized nations. The purpose of the current study was to determine the frequency of bone marrow involvement in our series of retinoblastoma patients and to investigate whether the use of four bone marrow aspirates (BMA) and two bone marrow biopsies (BMB) has greater sensitivity for the detection of metastatic disease compared to what has been previously reported. Methods Retrospective analysis of the charts of 54 patients with retinoblastoma was performed. We performed 265 BMA, 134 BMB (4 aspirates and 2 biopsies per evaluation), and 67 lumbar punctures (LPs) in 54 patients with retinoblastoma. Results There were no patients found with bone marrow or cerebrospinal fluid (CSF) involvement at the time of the initial diagnosis. Although no patient died of distant metastases, two patients developed metastatic disease at recurrence, involving the bone marrow and other sites. For these two patients all four aspirates and two biopsies were positive for disease at the time of recurrence. Conclusions Despite the use of four BMA and two BMB (as opposed to one bone marrow aspirate that is routinely performed in other centers), the detection of patients with metastatic disease was similar to what has been previously reported. Based on these data more aggressive evaluation of the bone marrow in retinoblastoma patients with clinically limited disease using four aspirates and two biopsies cannot be supported. © 2005 Wiley-Liss, Inc. [source] Acute lymphoblastic leukemia with the phenotype of a putative B-cell/T-cell bipotential precursorAMERICAN JOURNAL OF HEMATOLOGY, Issue 2 2004Lee Gong Lau Abstract Biphenotypic acute leukemias (BALs) are uncommon. Most are of myeloid-B-cell or myeloid-T-cell lineage. We report herein a 70-year-old man with an unusual acute leukemia where the blasts expressed both B- and T-lymphoid markers. He presented to us with an enlarging cutaneous tumor. The presenting peripheral blood and bone marrow aspirate showed 40% and 90% blasts, respectively, which were negative for the usual cytochemical stains. The flow cytometric analysis revealed that the blasts were positive for CD19, CD20, CD22, cytoplasmic (Cyt) CD79a, CD10, Cyt CD3, CD5, CD7, CD4, HLA-DR, TdT, and were negative for myeloid markers. According to the scoring system from the European Group for the Immunological Characterization of Acute Leukaemias (EGIL), this case was an unequivocal B-cell/T-cell BAL. Conventional cytogenetic analysis revealed 46XY [t(4;11)(q31;q13), add(8)(q24), der(9)del(9)(p21)del(9)(q32q34), ,13, +mar] in all 25 metaphases analyzed. Fluorescence in situ hybridization (FISH) and polymerase chain reaction (PCR) for 11q23 rearrangements as well as t(9;22) were negative. PCR for both TCR- , and IgH gene analyses revealed polyclonal rearrangements. We postulate that this case of BAL might have arisen from the putative common lymphoid progenitor cell. Am. J. Hematol. 77:156,160, 2004. © 2004 Wiley-Liss, Inc. [source] A sporadic case of visceral leishmaniasis from Kocaeli, Turkey,APMIS, Issue 11 2006Case report Visceral and cutaneous leishmaniasis are endemic in the western and southeastern parts of Turkey. We report a sporadic case of visceral leishmaniasis from Kocaeli, which is not an endemic area. The patient, a 10-month-old male infant, had since birth never been outside the city. He was referred to our hospital with a one-month history of fever. Antibiotics were administered but fever persisted. There were Leishman bodies in the bone marrow aspirate, both in macrophages and in clusters among other cells. Immunofluorescence antibody test (IFAT) detected no antibodies in the mother. Liposomal amphotericin B was administered. Visceral leishmaniasis should be considered in the differential diagnosis of patients with persistant fever, hepatosplenomegaly and cytopenia, even in nonendemic areas. [source] Efficacy of Bone Marrow Mononuclear Cells to Promote Bone Regeneration Compared With Isolated CD34+ Cells From the Same Volume of AspirateARTIFICIAL ORGANS, Issue 7 2010Shinji Yasuhara Abstract Autologous bone marrow mononuclear cell (BMMNC) transplantation is currently an emerging clinical treatment in the orthopedic as well as cardiovascular fields. It is believed that the therapeutic effect of the BMMNCs is due to neovascularization enhanced by the CD34+ cells contained therein, which include endothelial progenitor cells. However, isolation of the CD34+ cell fraction for clinical application has many disadvantages such as cost and invasiveness related to cell mobilization with cytokine. To investigate whether a purification step is in fact necessary for bone regeneration, we separated BMMNCs, CD34+, and CD34 - cells from the same initial volume of rabbit bone marrow aspirates. We then transplanted them back into a femoral bone defect of the same rabbit together with atelocollagen gel and basic fibroblast growth factor (bFGF) and evaluated neovascularization and bone regeneration up to 8 weeks after transplantation. The greatest potential for neovascularization and bone regeneration medicated by cells from the same volume of bone marrow aspirate was found in the BMMNC group. Although purified CD34+ cells might be an ideal cell source, BMMNCs could be a practical and feasible cell source for bone regeneration in present clinical settings with limited cost, availability of materials, and technical issues for transplantation. [source] Development of a New Tissue-Engineered Sheet for Reconstruction of the StomachARTIFICIAL ORGANS, Issue 10 2009Masato Araki Abstract We have developed tissue-engineered digestive tracts composed of collagen scaffold and an inner silicon sheet and successfully used it to repair defects in parts of the esophagus, stomach, and small intestine. However, some improvements were demanded for clinical usage because the silicon sheet presented technical difficulties for suturing and endoscopic removal. New tissue-engineered sheet (New-sheet) was composed of a single-piece and reinforced collagen scaffold with biodegradable copolymer. One beagle dog was used to evaluate whether New-sheet could withstand suturing in comparison with native digestive tracts using a tensile tester. Seven beagle dogs had a 5-cm circular defect created in the stomach. New-sheet soaked with autologous peripheral blood or bone marrow aspirate was sutured to the gastric wall. Endoscopic, histological, and immunohistochemical assessment was performed to evaluate regeneration of the stomach up to 16 weeks. Tensile strength testing showed that the mucosal side of New-sheet had strength almost equivalent to the mucosa of the esophagus (P = 0.61). Endoscopically, regeneration of the mucosa started from the circumference after 4 weeks, but a small linear ulcer was still evident at 16 weeks. The regenerated stomach shrank by 60,80% of its original size and histologically showed villous mucosa and underlying dense connective tissue. Immunohistochemically, the regenerated area expressed ,-smooth-muscle actin but was negative for basic calponin, irrespective of the source of soaked blood. New-sheet shows sufficient strength for suturing, no dehiscence, and better biocompatibility for clinical use, although further examination will be necessary to create a functional digestive tract. [source] Toxic epidermal necrolysis secondary to angioimmunoblastic T-cell lymphomaAUSTRALASIAN JOURNAL OF DERMATOLOGY, Issue 3 2005Brad Jones SUMMARY A 67-year-old man presented with a history of lymphadenopathy, fevers and separate skin eruptions of erythrodermic spongiotic dermatitis initially and subsequent toxic epidermal necrolysis. Initial lymph node biopsies showed non-specific granulomatous changes, and skin biopsies and bone marrow aspirate were not diagnostic. His toxic epidermal necrolysis responded well to 3 days of intravenous immunoglobulin. The patient was discharged from hospital and reviewed regularly as an outpatient. Due to persisting lymphadenopathy, further lymph node biopsy led to the diagnosis of angioimmunoblastic T-cell lymphoma, a rare form of peripheral T-cell lymphoma with a poor prognosis. At the time of diagnosis his condition deteriorated rapidly and he died soon after. [source] Detection of tumor specific gene expression in bone marrow and peripheral blood from patients with small cell lung carcinomaCANCER, Issue 4 2003Masato Shingyoji M.D. Abstract BACKGROUND Small cell lung carcinoma (SCLC) has the propensity to grow rapidly and metastasize extensively. Detection of micro-dissemination of SCLC may have clinical relevance. For its detection, tumor-specific gene expressions were examined in peripheral blood and bone marrow aspirate from patients with SCLC. METHODS Expression of prepro-gastrin-releasing peptide (preproGRP), neuromedin B receptor (NMB-R) and gastrin-releasing peptide receptor (GRP-R) were examined by reverse transcriptase polymerase chain reaction (RT-PCR) in peripheral blood and bone marrow aspirate from 40 untreated patients with SCLC. Control samples consisted of peripheral blood samples from 5 patients with nonsmall cell lung cancer (NSCLC) and 20 healthy volunteers. RESULTS Positive rates of preproGRP, NMB-R, and GRP-R in bone marrow aspirate of patients with SCLC were 23% (9/40), 8% (3/40), and 10% (4/40), respectively. Those rates in peripheral blood were 11% (4/38), 5% (2/38), and 29% (11/38), respectively. Although GRP-R expression was detected in patients with NSCLC and in healthy volunteers, preproGRP and NMB-R expressions were not detected in patients with NSCLC and in healthy volunteers. All three gene expressions in bone marrow were more frequently observed in patients with bone marrow metastasis, accessed by biopsy, than in patients without. PreproGRP gene expression in bone marrow was also more frequent in patients with bone metastasis, accessed by bone scintigram, than in patients without, and was related to poorer survival. CONCLUSIONS Micro-dissemination of SCLC was detectable by RT-PCR of preproGRP and NMB-R, both specific for SCLC. These gene expressions in bone marrow may be related to disease extent and prognosis. Cancer 2003;97:1057,62. © 2003 American Cancer Society. DOI 10.1002/cncr.11108 [source] Persistence of myeloma protein for more than one year after radiotherapy is an adverse prognostic factor in solitary plasmacytoma of boneCANCER, Issue 5 2002Richard B. Wilder M.D. Abstract BACKGROUND Prognostic factors for solitary plasmacytoma of bone (SPB), whether measured before or after radiotherapy (RT), have not been established. The authors analyzed multiple factors for myeloma-free survival (MFS) and cause-specific survival (CSS) in SPB patients treated with RT alone. METHODS Between 1965 and 2000, 60 patients with carefully staged SPB were treated with RT alone at the M. D. Anderson Cancer Center. Patient ages ranged from 29,77 years (median, 54 years), and 75% of patients had a myeloma (M) protein in the blood and/or urine. No patients showed other lesions on skeletal survey or, in recent years, magnetic resonance imaging (MRI) of the spine; marrow aspirate was normal in all patients. Radiotherapy to the solitary lesion was given to a total dose of 30,70 Gy (median, 46 Gy). The authors analyzed the impact of multiple factors on MFS and CSS, including resolution v. persistence of M protein after RT, secretory v. nonsecretory disease at diagnosis, presence v. absence of an associated soft tissue mass on computed tomography or MRI scan, magnitude of serum M protein elevation at diagnosis, age, spinal v. nonspinal location, Karnofsky performance status, total RT dose, and tumor size. RESULTS Median follow-up was 7.8 years (range, 1.0,25.5 years). On multivariate analysis, persistence of M protein more than one year after RT was the only independent adverse prognostic factor for MFS (P = 0.005) and CSS (P = 0.04). Most patients with M protein that persisted for more than one year after RT were diagnosed with multiple myeloma within 2.2 years of treatment. CONCLUSIONS Patients with M protein that persists for more than one year after RT should be monitored frequently and considered for standard chemotherapy followed by intensive consolidation therapy when they either develop symptoms or show an increasing M protein level. Cancer 2002;94:1532,7. © 2002 American Cancer Society. DOI 10.1002/cncr.10366 [source] Aleukaemic leukaemia cutis presenting as a benign-appearing eruptionCLINICAL & EXPERIMENTAL DERMATOLOGY, Issue 2 2003T. P. Millard Summary A 68-year-old Caucasian male presented with a 5-week history of a widespread pruritic papular eruption. Histology from a papule on the left shoulder showed a dense dermal infiltrate of large mononuclear cells which were positive for leucocyte common antigen, KP1 and PGM1, with an MIB-1 proliferating fraction of 40%, diagnostic of acute monocytic (M5) leukaemia cutis. Full blood count revealed pancytopaenia but no blasts. Bone marrow aspirate showed reduced red cell precursors and 10% blasts, consistent with myelodysplastic syndrome (refractory anaemia with excess blasts). The patient was managed with a 3 unit transfusion of packed red cells, after which his skin eruption resolved within 6 weeks and his peripheral blood counts returned to normal. No chemotherapy was administered. In conclusion, leukaemia can present in the skin, the eruption may be nonspecific and it may precede systemic involvement by either myelodysplastic syndrome or acute leukaemia. [source] Flow cytometric differential of leukocyte populations in normal bone marrow: Influence of peripheral blood contamination1,CYTOMETRY, Issue 1 2009R. A. Brooimans Abstract Background: Availability of immunophenotypic reference values for the various leukocyte populations distributed in bone marrow may be helpful to recognize abnormal bone marrow development and, therefore, useful as first screening of individuals with suspected hematological malignancies or other hematopoietic disorders. Methods: A single tube four-color staining panel (CD66abce/CD14/CD45/CD34) together with a predefined gating strategy was utilized to immunologically differentiate the distribution of the major leukocyte populations in bone marrow aspirates of healthy donors. The sample-blood erythrocyte ratio was applied to assess the amount of blood contamination of marrow and account for this in the marrow value estimates. Results: The frequency of the major leukocyte populations in bone marrow of 134 normal donors were for granulocytes: mean, 69.4%; SD, 10.3%; monocytes: mean, 4.7%; SD, 2.3%; lymphocytes: mean, 18.3%; SD, 8.7%. The frequency of the immature cell population that included precursor cells of each of the cell lineages among other cell types were mean 5.0%; SD 2.2%. The mean percentage of CD34 positive cells was 1.5%; SD 0.7%. Our results showed further that the frequency of cell populations, of which the presence is restricted to the bone marrow (e.g., CD34+ progenitor cells), is influenced by the degree of peripheral blood admixture. Between the total immature cells and purity of the bone marrow, there was a significant positive correlation demonstrated, whereas a negative correlation was found between the percentages of both lymphocytes as monocytes and the purity of the bone marrow. Conclusions: With a single tube-staining panel, we obtained reference values for flow cytometric assessment of all relevant leukocyte populations present in bone marrow that can be used as a frame of reference for better recognition of individuals with abnormal hematopoiesis. In addition, we have demonstrated the influence of the degree of peripheral blood admixture in the bone marrow aspirates on those reference values. © 2008 Clinical Cytometry Society [source] C-Kit receptor (CD117) expression on myeloblasts and white blood cell counts in acute myeloid leukemiaCYTOMETRY, Issue 1 2004Jolanta Wo Abstract Background The c-Kit receptor is considered to play a crucial role in hematopoiesis. Induction of mobilization of hematopoietic cells in the bone marrow requires cooperative signaling through c-Kit and c-Kit ligand pathway, and these interactions are important in the retention of stem cells within the bone marrow. Therefore, we analyzed c-Kit density on the leukemic myeloblasts of patients with acute myeloid leukemia (AML) in relation to white blood cell count (WBC) in the peripheral blood. Methods Bone marrow aspirates collected from patients with AML and bone marrow aspirates and leukapheresis products after granulocyte colony-stimulating factor blood mobilization from adult volunteers were studied. To determine the level of c-Kit receptor expression, we applied quantitative (relative fluorescence intensity and antibody binding per cell) cytometric methods. Results Our data showed negative correlation between the level of c-Kit expression intensity on myeloblasts and the number of leukocytes in blood of AML patients. The c-Kit receptor density on myeloblasts in patients with low WBC was significantly stronger than that on myeloblasts in patients with high WBC. In the latter patient group, the density c-Kit receptor on myeloblasts was similar to that on CD34+ cells in mobilized peripheral blood. Conclusions The obtained data suggest an involvement of c-Kit receptor in the regulation of leukemic myeloblasts egress to the peripheral blood. © 2004 Wiley-Liss, Inc. [source] CD87 as a marker for terminal granulocytic maturation: Assessment of its expression during granulopoiesisCYTOMETRY, Issue 1 2003M. Tarek Elghetany Abstract Background Understanding the normal surface maturation pattern of granulocytes is essential for the recognition of abnormal patterns, which in turn may be of diagnostic or pathogenetic significance in disorders such as myelodysplastic syndromes and inherited bone marrow failure disorders. CD87 plays a role in cellular interaction, cell migration, and inflammatory response. Surface expression of this antigen has not been adequately studied on bone marrow granulocytes, and the small number of previous studies has provided conflicting data. Methods Bone marrow aspirates from 11 control subjects were studied by flow cytometry and a lysed whole blood technique to compare surface expression of CD87 on marrow granulocytes with those of CD11b, CD16, CD35, and CD10, which are expressed at the myelocyte, metamyelocyte, band, and segmented stage of neutrophilic development, respectively. Four sorting experiments of CD87+ granulocytes were also performed. Results Our study showed no statistical difference between surface expression of CD35 and CD87 (P > 0.3), whereas significant differences existed between CD87 and the other antibodies (P < 0.004). Sorting experiments showed that more than 80% of CD87+ cells were bands and segmented neutrophils. Dual staining for CD87 and CD35 showed that most CD87+ granulocytes coexpress CD35. Conclusions CD87 is expressed on granulocytes at the band and segmented neutrophil stage of development and can be used to study normal and abnormal granulopoiesis. Cytometry Part B (Clin. Cytometry) 51B:9,13, 2003. © 2002 Wiley-Liss, Inc. [source] Cell microarray platform for anticancer drug development,DRUG DEVELOPMENT RESEARCH, Issue 5 2007Min-Jung Lee Abstract Pharmacodynamic assessment of whether a drug has interacted with and modified its target is an essential component of molecularly targeted clinical trials. Although many trials are written with the intent to assess tumor biopsies, if available, thus far the great majority of early drug trials have used peripheral blood mononuclear cells (PBMC) as a tumor surrogate. Typically, PBMC are studied by low-throughput techniques such as Western blot. We present the use of a cell-based tissue microarray for assessment of anticancer drug activity in vivo. We demonstrate the utility of this technique for analysis of protein hyperacetylation in response to treatment with the histone deacetylase inhibitor, SNDX-275 in PBMC treated in vitro and in PBMC and bone marrow aspirates from patients in Phase I clinical trials with SNDX-275. We demonstrate that the cell microarray can be used to measure drug response in a high-throughput manner, allowing analysis of an entire trial on one or two glass slides. The cell microarray technique brings the advantages of the tissue microarray platform to the pharmacodynamic assessment of single cells, such as those isolated from bone marrow aspirates, fine needle aspirates, or malignant effusions, and to analysis of PBMC, the most commonly studied surrogate in oncology trials. Drug Dev Res 68:226,234, 2007. Published 2007 Wiley-Liss, Inc. [source] BSc2118 is a novel proteasome inhibitor with activity against multiple myelomaEUROPEAN JOURNAL OF HAEMATOLOGY, Issue 2 2010Jan Sterz Abstract Objectives:, The ubiquitin,proteasome system emerged as a new therapeutic target in cancer treatment. The purpose of this study was to elucidate the effects of the novel proteasome inhibitor BSc2118 on t(4;14) positive and negative multiple myeloma (MM) cells and normal peripheral blood mononuclear cells (PBMNC). Methods:, Human MM cell lines OPM-2, RPMI-8226, and U266 and primary MM cells from bone marrow aspirates were exposed to BSc2118. Cytotoxicity levels were evaluated using the MTT-test. BSc2118-induced apoptosis was analyzed by annexin-V assay. Further methods used included proteasomal activity determination, cell cycle analysis, western blot, and transcription factor assays. Results:, In OPM-2, RPMI-8226, U266 cell lines and primary MM cells, BSc2118 caused dose-dependent growth inhibitory effects. After 48 h, dose-dependent apoptosis occurred both in cell lines and primary myeloma cells irrespective of t(4;14). A significant G2-M cell cycle arrest occurred after 24 h. Furthermore, we observed a marked inhibition of intracellular proteasome activity, an increase in intracellular p21 levels, and an inhibition of NF-,B activation. The toxicity against PBMNC remained low, suggesting a broad therapeutic range of this agent. Conclusion:, Taken together, BSc2118 shows significant antimyeloma activity and may be considered as a promising agent in cancer drug development. [source] Detection of bone marrow-disseminated breast cancer cells using an RT-PCR assay of MUC5B mRNAINTERNATIONAL JOURNAL OF CANCER, Issue 4 2003Nora Berois Abstract The evaluation of disseminated epithelial tumor cells in breast cancer patients has generated considerable interest due to its potential association with disease recurrence. Our work was performed to analyze the usefulness of 5 mucin genes expression (MUC2, MUC3, MUC5B, MUC6 and MUC7), using RT-PCR assays, to detect disseminated cancer cells in patients with operable breast cancer. The highest frequencies of positive RT-PCR tests in breast tumor extracts were observed for MUC5B (7/15) and MUC7 (5/12). The best specificity, negative results on all peripheral blood mononuclear (PBMN) cell samples from healthy donors, were shown for MUC2, MUC5B and MUC6 RT-PCR assays. Thus, we selected MUC5B as a target gene for further evaluation. Using a nested RT-PCR, MUC5B mRNA transcripts were detected in 16/31 primary breast tumors (but not in 36 samples of normal PBMN cells) and in the human MCF-7 breast cancer cell line but not in BT20, MDA, T47D and ZR-75 breast cancer cell lines, indicating that MUC5B mRNA is expressed in a population of breast cancer cells. Using this method, 9/46 patients (19.5%) who underwent curative surgery showed positive MUC5B mRNA in bone marrow aspirates obtained prior to surgery, including 5/24 patients (20.8%) with stage I or II breast cancer, without histopathologic lymph node involvement. These results indicate that MUC5B mRNA could be a specific marker applicable to the molecular diagnosis of breast cancer cell dissemination. A comparative evaluation between MUC5B mRNA, cytokeratin 19 (CK19) mRNA and carcinoembryonic antigen (CEA) mRNA in all bone marrow aspirates suggests a putative complementation for molecular detection of disseminated carcinoma cells. Considering that breast cancer is characterized by a great phenotypic heterogeneity, the use of multimarker approach could contribute to tumor cell detection in bone marrow and blood. © 2002 Wiley-Liss, Inc. [source] Image analysis systems for the detection of disseminated breast cancer cells on bone-marrow cytospinsJOURNAL OF CLINICAL LABORATORY ANALYSIS, Issue 3 2005Sven Becker Abstract We assessed the accuracy of automated cell imaging systems when compared to manual evaluation of cytospin slides in determining the presence of cytokeratin-positive, disseminated breast cancer cells in bone marrow aspirates. A total of 298 cytospin slides of bone marrow aspirates were first evaluated by individual screening by one expert immunocytologist. Subsequently, all 298 slides were evaluated by the Automated Cell Imaging System (ACIS) by ChromaVisionÔ. Two separate analysis cycles were performed using ACIS. The results of the two ACIS analysis cycles were almost identical: in 293 out of 298 samples (98.3%), identical numbers of disseminated breast cancer cells were detected. In the remaining five samples (1.7%), the result of the two ACIS analysis cycles differed by only one tumor cell. By using the manual technique, 120 cytospin samples were found to be positive. ACIS was able to detect additional tumor cells in 64 cases. Not once did ACIS miss tumor cells when compared to the manual technique. Using ACIS, we were able to determine the bone marrow status of patients with nonmetastatic breast cancer faster, with greater accuracy, and with greater reproducibility than with the manual technique. J. Clin. Lab. Anal. 19:115,119, 2005. © 2005 Wiley-Liss, Inc. [source] Hematologic aspects of myeloablative therapy and bone marrow transplantationJOURNAL OF CLINICAL LABORATORY ANALYSIS, Issue 2 2005Roger S. Riley Abstract The transplantation of bone marrow cells or isolated hematopoietic stem cells from the bone marrow or peripheral blood is a widely utilized form of therapy for patients with incurable diseases of the hematopoietic and immune systems. Successful engraftment of the transplanted stem cells in an adequately prepared recipient normally leads to bone marrow reconstitution over a period of several weeks, accompanied by more gradual reconstitution of the immune system. Since the recipient is profoundly ill during the initial treatment period, laboratory data is critical for monitoring engraftment, detecting residual/recurrent disease, and identifying problems that may delay bone marrow reconstitution or lead to other medical complications. Accurate blood cell counts are imperative, and most bone marrow transplantation patients undergo periodic monitoring with bone marrow aspirates and biopsies with cytogenetic, molecular, and multiparametric flow cytometric studies. The potential complications of bone marrow transplantation include engraftment failure and delayed engraftment, infection, residual bone marrow disease, acute and chronic graft versus host disease, myelofibrosis, therapy-related acute leukemia, post-transplant lympho-proliferative disorders, and toxic myelopathy. J. Clin. Lab. Anal. 19:47,79, 2005. © 2005 Wiley-Liss, Inc. [source] Possible automatic cell classification of bone marrow aspirate using the CELL-DYN 4000® automatic blood cell analyzerJOURNAL OF CLINICAL LABORATORY ANALYSIS, Issue 2 2002Ryousuke Yamamura Abstract In clinical hematology, the demand for bone marrow aspiration testing is increasing. However, conventional automatic blood cell analyzers cannot completely analyze erythroblasts, and evaluation has mainly been performed by visual examination (the microscopic method). Using the CELL-DYN 4000® automatic blood cell analyzer (CD4000) (Abbott Laboratories, North Chicago, IL), specific recognition and classification of erythroblasts by DNA staining is possible. In the present study, using bone marrow blood collected from normal subjects and patients with hematological malignancy, we classified cells by the microscopic method and with the CD4000, and compared the results. Good correlations were found for total nucleated cell count (TNCC), neutrophils, lymphocytes, erythroblasts, and the myeloid series to erythroid series (M/E) ratio. It is possible to detect blasts that emerge in patients with hematological malignancy using the blast flag system installed on the CD4000. Since all of the items can be analyzed in about 80 sec with the CD4000, cells in bone marrow aspirates can be classified faster with this apparatus than by the microscopic method. Therefore, analysis of bone marrow aspirates with this apparatus appears to be very useful not only for laboratory testing but also for clinical screening. J. Clin. Lab. Anal. 16:86,90, 2002. © 2002 Wiley-Liss, Inc. [source] Rotating three-dimensional dynamic culture of adult human bone marrow-derived cells for tissue engineering of hyaline cartilageJOURNAL OF ORTHOPAEDIC RESEARCH, Issue 4 2009Shinsuke Sakai Abstract The method of constructing cartilage tissue from bone marrow-derived cells in vitro is considered a valuable technique for hyaline cartilage regenerative medicine. Using a rotating wall vessel (RWV) bioreactor developed in a NASA space experiment, we attempted to efficiently construct hyaline cartilage tissue from human bone marrow-derived cells without using a scaffold. Bone marrow aspirates were obtained from the iliac crest of nine patients during orthopedic operation. After their proliferation in monolayer culture, the adherent cells were cultured in the RWV bioreactor with chondrogenic medium for 2 weeks. Cells from the same source were cultured in pellet culture as controls. Histological and immunohistological evaluations (collagen type I and II) and quantification of glycosaminoglycan were performed on formed tissues and compared. The engineered constructs obtained using the RWV bioreactor showed strong features of hyaline cartilage in terms of their morphology as determined by histological and immunohistological evaluations. The glycosaminoglycan contents per µg DNA of the tissues were 10.01,±,3.49 µg/µg DNA in the case of the RWV bioreactor and 6.27,±,3.41 µg/µg DNA in the case of the pellet culture, and their difference was significant. The RWV bioreactor could provide an excellent environment for three-dimensional cartilage tissue architecture that can promote the chondrogenic differentiation of adult human bone marrow-derived cells. © 2008 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 27: 517,521, 2009 [source] The effect of oxygen tension on the in vitro assay of human osteoblastic connective tissue progenitor cellsJOURNAL OF ORTHOPAEDIC RESEARCH, Issue 10 2008Sandra M. Villarruel Abstract Connective tissue progenitors (CTPs) are defined as the heterogeneous set of stem and progenitor cells that reside in native tissues and are capable of proliferation and differentiation into one or more connective tissue phenotypes. CTPs play important roles in tissue formation, repair, and remodeling. Therefore, in vitro assays of CTP prevalence and biological potential have important scientific and clinical relevance. This study evaluated oxygen tension as an important variable in optimizing in vitro conditions for quantitative assays of human CTPs. Bone marrow aspirates were collected from 20 human subjects and cultured using established medium conditions at ambient oxygen tensions of 1, 5, 10, and 20%. Colony-forming efficiency (CFE), proliferation, and colony density were assessed. CFE and proliferation were greatest at 5% O2. Traditional conditions using atmospheric oxygen tension (20% O2) reduced CFE by as much as 32%. CFE and proliferation at 1% O2 were less than 5% O2 but comparable to that seen at 20% O2, suggesting that CTPs are relatively resilient under hypoxic conditions, a fact that may be relevant to their function in wound repair and their potential use in tissue engineering applications involving transplantation into settings of moderate to severe hypoxia. These data demonstrate that optimization of quantitative assays for CTPs will require control of oxygen tension. © 2008 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 26:1390,1397, 2008 [source] More aggressive bone marrow screening in retinoblastoma patients is not indicated: The memorial Sloan-Kettering cancer center experience,PEDIATRIC BLOOD & CANCER, Issue 1 2006Stergios Zacharoulis MD Abstract Background Bone marrow involvement in retinoblastoma patients is rare in the more industrialized nations. The purpose of the current study was to determine the frequency of bone marrow involvement in our series of retinoblastoma patients and to investigate whether the use of four bone marrow aspirates (BMA) and two bone marrow biopsies (BMB) has greater sensitivity for the detection of metastatic disease compared to what has been previously reported. Methods Retrospective analysis of the charts of 54 patients with retinoblastoma was performed. We performed 265 BMA, 134 BMB (4 aspirates and 2 biopsies per evaluation), and 67 lumbar punctures (LPs) in 54 patients with retinoblastoma. Results There were no patients found with bone marrow or cerebrospinal fluid (CSF) involvement at the time of the initial diagnosis. Although no patient died of distant metastases, two patients developed metastatic disease at recurrence, involving the bone marrow and other sites. For these two patients all four aspirates and two biopsies were positive for disease at the time of recurrence. Conclusions Despite the use of four BMA and two BMB (as opposed to one bone marrow aspirate that is routinely performed in other centers), the detection of patients with metastatic disease was similar to what has been previously reported. Based on these data more aggressive evaluation of the bone marrow in retinoblastoma patients with clinically limited disease using four aspirates and two biopsies cannot be supported. © 2005 Wiley-Liss, Inc. [source] Reduced chondrogenic and adipogenic activity of mesenchymal stem cells from patients with advanced osteoarthritisARTHRITIS & RHEUMATISM, Issue 3 2002J. Mary Murphy Objective Mesenchymal stem cells (MSCs) are resident in the bone marrow throughout normal adult life and have the capacity to differentiate along a number of connective tissue pathways, among them bone, cartilage, and fat. To determine whether functionally normal MSC populations may be isolated from patients with advanced osteoarthritis (OA), we have compared cells from patients undergoing joint replacement with cells from normal donors. Cell populations were compared in terms of yield, proliferation, and capacity to differentiate. Methods MSCs were prepared from bone marrow aspirates obtained from the iliac crest or from the tibia/femur during joint surgery. In vitro chondrogenic activity was measured as glycosaminoglycan and type II collagen deposition in pellet cultures. Adipogenic activity was measured as the accumulation of Nile Red O-positive lipid vacuoles, and osteogenic activity was measured as calcium deposition and by von Kossa staining. Results Patient-derived MSCs formed colonies in primary culture that were characteristically spindle-shaped with normal morphology. The primary cell yield in 36 of 38 cell cultures from OA donors fell within the range found in cultures from normal donors. However, the proliferative capacity of patient-derived MSCs was significantly reduced. There was a significant reduction in in vitro chondrogenic and adipogenic activity in cultures of patient-derived cells compared with that in normal cultures. There was no significant difference in in vitro osteogenic activity. There was no decline in chondrogenic potential with age in cells obtained from individuals with no evidence of OA. Conclusion These results raise the possibility that the increase in bone density and loss of cartilage that are characteristic of OA may result from changes in the differentiation profile of the progenitor cells that contribute to the homeostatic maintenance of these tissues. [source] Efficacy of Bone Marrow Mononuclear Cells to Promote Bone Regeneration Compared With Isolated CD34+ Cells From the Same Volume of AspirateARTIFICIAL ORGANS, Issue 7 2010Shinji Yasuhara Abstract Autologous bone marrow mononuclear cell (BMMNC) transplantation is currently an emerging clinical treatment in the orthopedic as well as cardiovascular fields. It is believed that the therapeutic effect of the BMMNCs is due to neovascularization enhanced by the CD34+ cells contained therein, which include endothelial progenitor cells. However, isolation of the CD34+ cell fraction for clinical application has many disadvantages such as cost and invasiveness related to cell mobilization with cytokine. To investigate whether a purification step is in fact necessary for bone regeneration, we separated BMMNCs, CD34+, and CD34 - cells from the same initial volume of rabbit bone marrow aspirates. We then transplanted them back into a femoral bone defect of the same rabbit together with atelocollagen gel and basic fibroblast growth factor (bFGF) and evaluated neovascularization and bone regeneration up to 8 weeks after transplantation. The greatest potential for neovascularization and bone regeneration medicated by cells from the same volume of bone marrow aspirate was found in the BMMNC group. Although purified CD34+ cells might be an ideal cell source, BMMNCs could be a practical and feasible cell source for bone regeneration in present clinical settings with limited cost, availability of materials, and technical issues for transplantation. [source] Telomerase activity in disseminated prostate cancer cellsBJU INTERNATIONAL, Issue 6 2006JESCO PFITZENMAIER OBJECTIVE To analyse telomerase activity in disseminated prostate cancer cells isolated from bone marrow aspirates taken from men with localized prostate cancer before radical prostatectomy (RP). PATIENTS AND METHODS Disseminated epithelial prostate cancer cells were isolated from bone marrow aspirates from 69 men with localized prostate cancer before RP, by magnetic column-chromatography enrichment, followed by isolation of fluorescently labelled epithelial cells by micropipetting. We used pools of 10 non-epithelial bone marrow cells after tumour cell enrichment as control samples. These pure cell pools were tested for the presence of telomerase activity. RESULTS In all, 49 of the patient samples contained disseminated prostate cancer cells. Homogeneous pools of 10 cells were obtained from 35 of these; 49% of the 35 specimens showed telomerase activity, whereas all five control samples did not. Telomerase activity in the 35 samples was not significantly associated with Gleason score, preoperative prostate-specific antigen level, tumour stage, or surgical margin status. Follow-up is continuing to assess an association with disease recurrence. CONCLUSION This work shows the feasibility of isolating disseminated cancer cells for analysing individual or pooled cells. Compared to tissue staining, where telomerase is detected in 80,90% of samples, we found lower rates of telomerase activity in the disseminated tumour cells (49%). Telomerase-negative cells might provide information about cell dormancy, as telomerase is a marker of cell proliferation in immortal and cancer cells. Telomerase-positive cells might predict early disease recurrence, but a longer follow-up is needed to test this possibility. [source] Post-kala-azar dermal leishmaniasis in the Sudan: clinical presentation and differential diagnosisBRITISH JOURNAL OF DERMATOLOGY, Issue 1 2000E.E. Zijlstra Post-kala-azar dermal leishmaniasis (PKDL) is a common complication following kala-azar (visceral leishmaniasis). In a prospective study in a village in the endemic area for kala-azar in the Sudan, 105 of 183 (57%) kala-azar patients developed PKDL. There was a significantly higher PKDL rate (69%) in those who received inadequate and irregular treatment of kala-azar than in those who were treated with stibogluconate 20 mg kg,1 daily for 15 days (35%). The group of patients who developed PKDL did not differ from those who did not develop PKDL with regard to age and sex distribution, reduction in spleen size, and conversion in the leishmanin skin test (LST). In a clinical study, 416 PKDL patients were analysed and divided according to grade of severity. Severe PKDL was more frequent in younger age groups (P < 0·001); there was an inverse correlation between grade and conversion in the LST (P < 0·01). In 16% of patients tested, parasites were demonstrated in inguinal lymph node or bone marrow aspirates, indicating still visceral disease (para-kala-azar dermal leishmaniasis); there was no correlation between the presence of parasites and grade of severity. Conversion rates in the LST were lower than in those who did not have demonstrable parasites (11% and 37%, respectively; P < 0·01). In the absence of reliable and practical diagnostic tests, PKDL may be diagnosed on clinical grounds and differentiated from other conditions, of which miliaria rubra was the most common. Differentiation from leprosy was most difficult. [source] | ||||||||||||||||||||||||||||