scholarly journals Implementation of Flow Cytometric Monocyte Repartition Assay for Assessment of Chronic Myelomonocytic Leukaemia (CMML) in Bone Marrow

Blood ◽  
2020 ◽  
Vol 136 (Supplement 1) ◽  
pp. 3-4
Author(s):  
Sharon A Allen ◽  
Eugene Ng ◽  
Suzanne Edwards ◽  
Uwe H Hahn ◽  
Tatjana Banovic ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Myeloid Leukaemia ◽  
Peripheral Blood ◽  
Linear Regression Analysis ◽  
Myeloproliferative Neoplasm ◽  
P Value ◽  
Monocyte Subsets ◽  
Roc Curve Analysis ◽  
Flow Cytometric ◽  
Significant Difference

Background: CMML is a myelodysplastic/myeloproliferative overlap syndrome associated with persistent monocytosis. Previous studies have shown a characteristic phenotype of peripheral blood (PB) monocytes in CMML that may have diagnostic utility. There is expansion of the classical monocyte (Mo1) population (CD14+CD16-) with concomitant reduction in the intermediate (Mo2; CD14+16+) and non-classical (Mo3; CD14-16+) populations. It is unclear whether an increased Mo1 population or a reduced Mo2/Mo3 population is a better discriminator in detecting CMML by flow cytometry. Diagnostic thresholds for these populations have most often been established by comparison with normal donors using PB samples. The objective of this study was to determine the optimal cut-off limits for flow cytometric monocyte repartition (FMR) in bone marrow (BM) samples and to assess the diagnostic accuracy of this method in patients undergoing BM evaluation for other causes of monocytosis using a novel gating strategy. Patients/Methods: 71 BM aspirates were analysed comprising 35 confirmed CMML (21 proliferative CMML , 14 dysplastic CMML), 13 other myeloid neoplasm (MN) disease controls and 23 normal bone marrow aspirates. The MN cohort was identified through local samples with a known peripheral blood monocytosis of ≥1.0x109 and peripheral blast count of <20% not fulfilling criteria for CMML. The MN cohort comprised of 4 cases of chronic myeloid leukaemia (CML), 2 cases of myelodysplasia/myeloproliferative neoplasm unspecified (MDS/MPN-U), two cases of atypical CML (aCML), 2 cases of primary myelofibrosis, 1 case of juvenile myelomonocytic leukaemia (JMML), 1 case of chronic neutrophilic leukaemia (CNL) and 1 case of therapy-related acute myeloid leukaemia (T-AML). The median age of CMML patients within our cohort was 74 years (range 44- 89 years) and 77% were male. Stored flow cytometric data were reanalysed to assess monocyte subsets. After initial removal of doublets and non-cellular events, sequential gating selected a CD45+CD33bright population with low side scatter to select monocytes and exclude granulocytes. The lymphocyte gate as determined on the CD45 vs CD33 plot was used to establish a threshold for CD16 positivity. Immature double negative monocytes (CD14-CD16-) were excluded as in published gating strategies. Results: Linear regression analysis of the percentage of both the Mo1 and Mo2 populations demonstrated a significant difference between the CMML, MN, and normal cohorts (global P<0.0001). Post hoc analysis demonstrated a significant increase in the Mo1% fraction (mean 92.5%; 95% Confidence Interval [CI]: 88.1%, 96.9%) in CMML cases when compared to the normal BM cohort (mean 63.5%; 95% CI: 58.1%, 69.0%; comparison P <0.0001) and the MN cohort (mean 79.9%; 95% CI: 72.7%,87.2%; comparison P = 0.004). Conversely, the Mo2% was significantly decreased (mean 6.8% 95% CI: 2.7%, 10.9%) in CMML cases when compared to the normal cohort (mean 33.0%; 95% CI: 27.9%; 38.1%; comparison P value <0.0001) and the MN cohort (mean 17.6%; 95% CI 10.8%; 24.3%; comparison P value 0.008). Receiver Operator Curve (ROC) curve analysis of the Mo1 population demonstrated an area under the curve (AUC) of 0.8738 (95% CI: 0.7897, 0.9579; P<0.0001) with a marginally lower AUC for Mo2% (Figure 1A). Priority was given to selecting a cut-off value with higher specificity, considering that the BM examination is not a screening test. A cut-off value for Mo1% of >88.5% (Figure 1B) gave a sensitivity of 77.1% (95% CI: 60.98%, 87.93%) and specificity of 88.9% (95% CI: 74.69%, 95.59%) (Figure 1A). A Mo2% cut-off value of <12.25% (Figure 1C) gave a sensitivity of 78.8% (95% CI: 62.25%, 89.32%) with a specificity of 80.6% (95% CI: 64.97%, 90.25%) (Figure 1A). Eight patients with CMML were misclassified by the Mo1/Mo2 criteria. Conclusion: Our study confirms that analysis of monocyte subsets on BM samples provides good discrimination of CMML cases when compared to normal controls. There was significant overlap with other MN samples. FMR may be a useful adjunct to BM morphology in cases with subtle dysplasia. Further study is required to understand the overlap in monocyte phenotype between CMML and other myeloid neoplasms. Disclosures Allen: Amgen: Other: Travel Support; Roche: Other: Travel Support; Novartis: Other: Travel Support. Hahn:Roche: Honoraria; Astra Zeneca: Honoraria.

Peripheral Blood Stem Cells vs Bone Marrow for Matched Sibling Transplant in AML and ALL in First Remission.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 3321-3321 ◽  
Author(s):  
Mark Kirschbaum ◽  
M. O’Donnell ◽  
R. Spielberger ◽  
R. Bhatia ◽  
V. Pullarkat ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Platelet Count ◽  
Peripheral Blood ◽  
Acute Gvhd ◽  
P Value ◽  
Peripheral Blood Stem Cells ◽  
Significant Difference ◽  
Blood Stem Cells ◽  
Matched Sibling

Abstract We report a retrospective comparison of peripheral blood (PBSC) versus bone marrow derived stem cells for allogeneic transplant in a cohort of similarly treated patients based on eight year data from a single institution. Over the period 1995 to 2003, a total of 151 patients with AML or ALL in first CR underwent allogeneic matched sibling donor transplant, 98 AML patients and 53 with ALL. The source of stem cells was bone marrow in 69, G-CSF mobilized peripheral blood stem cells (PBSC) in 82. Of the AML patients, 40 (58%) received marrow, 58 (71%) received PBSC. Of the ALL patients, 29 (42%) received marrow, 24 (29%) got PBSC. The age of patients receiving marrow was 32.7 ± 13.6, and for those getting PBSC was 36.1 ± 16.1. The conditioning regimen was fully ablative (TBI/VP-16) in all but five patients (Fludarabine/Melphalan in 1 marrow and 4 PBSC pts). GVHD prophylaxis was with cyclosporine/methotrexate in patients receiving ablative conditioning, and CSP/MMF in the flu/mel patients. The time to achieving an ANC >1000 was significantly shorter (P value=.0001) in the PBSC group, 18.2 ±6.6 days vs. 27.2 ±48.7 in the marrow group. Time to achieving a platelet count greater than 25K was significantly shorter (P value=2x10−11) for the PBSC group, 20.9 ± 8.5 vs. 31.8 ± 11.2 for marrow. This was true as well for time to reach platelet counts greater than 100K (P value=10−8), 29.7 ± 28 days for PBSC, 55.8 ±55.6 for marrow. In fact there was a significant increase (P=.0001) in the number of patients, 29 (42%) of marrow recipients vs 11 (13%) of PBSC, who did not at any time achieve a platelet count of >100K. There was no significant difference in survival at 2 years or 4 years between marrow or PBSC-Kaplan-Meier survival probability at 2 and 4 years respectively for AML was.66 and.54 for marrow, and.67 and.54 for PBSC. In ALL, the 2 and 4 year K-M survival probability was.72 and.65 for marrow;.69 and.69 for PBSC recipients. There was no significant difference in acute GVHD, with grade I-II described in 19 (28%) of the marrow pts, 29 (35%) in the PBSC pts, and grade II-IV in 8 (12%) marrow pts and 11 (13%) receiving PBSC (p value =.57), or chronic GVHD (P value=.62). Interestingly, in a subset analysis of patients who were diagnosed with acute GVHD, there was a significant difference in disease free survival at 5 years (see graph), suggesting that there is a qualitative difference between acute GVHD observed as a result of bone marrow versus PBSC as a source of stem cells. Figure Figure

Over-expression of IL-6 coding gene in the peripheral blood of migraine with aura patients

2021 ◽  
pp. 1-5
Author(s):  
Mahdi Ramezani ◽  
Alireza Komaki ◽  
Mohammad Mahdi Eftekharian ◽  
Mehrdokht Mazdeh ◽  
Soudeh Ghafouri-Fard
Keyword(s):  
Migraine With Aura ◽  
Peripheral Blood ◽  
Healthy Subjects ◽  
Migraine Without Aura ◽  
P Value ◽  
Healthy Controls ◽  
Over Expression ◽  
Significant Difference ◽  
Male Patients ◽  
Years Lived With Disability

Migraine is a common disorder which is placed among the top ten reasons of years lived with disability. Cytokines are among the molecules that contribute in the pathophysiology of migraine. In the current study, we evaluated expression levels of IL-6 coding gene in the peripheral blood of 120 migraine patients (54 migraine without aura and 66 migraine with aura patients) and 40 healthy subjects. No significant difference was detected in expression of IL-6 between total migraine patients and healthy controls (Posterior beta = 0.253, P value = 0.199). The interaction effect between gender and group was significant (Posterior beta =-1.274, P value = 0.011), therefore, we conducted subgroup analysis within gender group. Such analysis revealed that while expression of this gene is not different between male patients and male controls (Posterior beta =-0.371, P value > 0.999), it was significantly over-expressed in female patients compared with female controls (Posterior beta = 0.86, P= 0.002). Expression of IL-6 was significantly higher in patients with aura compared with controls (Posterior beta = 0.63, adjusted P value = 0.019). However, expression of this cytokine coding gene was not different between patients without aura and healthy subjects (Posterior beta = 0.193, adjusted P value = 0.281). Therefore, IL-6 might be involved in the pathophysiology of migraine among females and migraine with aura among both sexes.

scholarly journals Clinical observations of bone marrow transfusion for promoting bone marrow reconstruction after chemotherapy for AIDS-related lymphoma

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Yixuan Liu ◽  
Suhong Xie ◽  
Lei Li ◽  
Yanhui Si ◽  
Weiwei Zhang ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Immune System ◽  
T Lymphocytes ◽  
Peripheral Blood ◽  
Autologous Bone Marrow ◽  
Control Group ◽  
Peripheral Vein ◽  
Significant Difference ◽  
Autologous Bone ◽  
Aids Related Lymphoma

Abstract Background This study investigates the effect of autologous bone marrow transfusion (BMT) on the reconstruction of both bone marrow and the immune system in patients with AIDS-related lymphoma (ARL). Methods A total of 32 patients with ARL participated in this study. Among them, 16 participants were treated with conventional surgery and chemotherapy (control group) and the remaining 16 patients were treated with chemotherapy followed by autologous bone marrow transfusion via a mesenteric vein (8 patients, ABM-MVI group) or a peripheral vein (8 patients, ABM-PI group). Subsequently, peripheral blood and lymphocyte data subsets were detected and documented in all patients. Results Before chemotherapy, no significant difference in indicators was observed between three groups of ARL patients. Unexpectedly, 2 weeks after the end of 6 courses of chemotherapy, the ABM-MVI group, and the ABM-PI group yielded an increased level of CD8+T lymphocytes, white blood cells (WBC), and platelet (PLT) in peripheral blood in comparison to the control group. Notably, the number of CD4+T lymphocytes in the ABM-PI group was significantly higher than that in the other two groups. Additionally, no significant difference in haemoglobin levels was observed before and after chemotherapy in both the ABM-MVI and ABM-PI groups, while haemoglobin levels in the control group decreased significantly following chemotherapy. Conclusions Autologous bone marrow transfusion after chemotherapy can promote the reconstruction of both bone marrow and the immune system. There was no significant difference in bone marrow recovery and reconstruction between the mesenteric vein transfusion group and the peripheral vein transfusion group.

scholarly journals FDG-PET vs. chemical shift MR imaging in differentiating intertrabecular metastasis from hematopoietic bone marrow hyperplasia

2021 ◽  
Author(s):  
Nozomi Oki ◽  
Yohei Ikebe ◽  
Hirofumi Koike ◽  
Reiko Ideguchi ◽  
Daisuke Niino ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Chemical Shift ◽  
Fdg Pet ◽  
Standardized Uptake Value ◽  
Roc Curve Analysis ◽  
Cutoff Value ◽  
Significant Difference ◽  
Histopathological Studies ◽  
Fdg Pet Ct ◽  
Hematopoietic Bone Marrow

Abstract Purpose To evaluate the utility of SUVmax on FDG-PET and chemical shift imaging (CSI) on MRI in the differentiation of intertrabecular metastasis (ITM) from hematopoietic bone marrow hyperplasia (HBMH). Patients and methods We retrospectively evaluated 54 indeterminate focal bone marrow lesions in 44 patients detected on FDG-PET. The lesions were assigned to the metastasis group (M group, 29 lesions of 24 patients) and the non-metastasis group (non-M group, 25 lesions of 20 patients) based on the follow-up or the histopathological studies. The lesions were assessed with the maximum standardized uptake value (SUVmax) on FDG-PET CT images and signal change ratio (SCR) on CSI. Results The median SUVmax were 5.62 and 2.91; the median SCR were − 0.08 and − 34.8 in M and non-M groups respectively, with significant difference (p < 0.001). With ROC curve analysis, the optimal cutoff value of SUVmax was 4.48 with a sensitivity of 72.4%, a specificity of 100%, and AUC of 0.905. The cutoff value of SCR was − 6.15 with a sensitivity of 82.8%, a specificity of 80%, and AUC of 0.818. Conclusion FDG-PET and CSI on MRI are useful in distinguishing ITM from HBMH. Though their sensitivities are similar, the specificity of FDG-PET was higher than that of MRI.

Immunophenotypic Study of Basophils by Multiparameter Flow Cytometry

2008 ◽  
Vol 132 (5) ◽  
pp. 813-819
Author(s):  
Xiaohong Han ◽  
Jeffrey L. Jorgensen ◽  
Archana Brahmandam ◽  
Ellen Schlette ◽  
Yang O. Huh ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Chronic Myelogenous Leukemia ◽  
Peripheral Blood ◽  
Myelogenous Leukemia ◽  
Multiparameter Flow Cytometry ◽  
Color Flow ◽  
Aberrant Expression ◽  
Flow Cytometric ◽  
Hla Dr

Abstract Context.—The immunophenotypic profile of basophils is not yet fully established, and the immunophenotypic changes in chronic myelogenous leukemia are not fully characterized. Objective.—To establish a comprehensive immunophenotypic spectrum of normal basophils and to assess the range of immunophenotypic aberrations of basophils in chronic myelogenous leukemia. Design.—Using 4-color flow cytometry, we compared the immunophenotypic profile of basophils in peripheral blood or bone marrow samples from 20 patients with no evidence of neoplasia to basophils from 15 patients with chronic myelogenous leukemia. Results.—Basophils in control cases were all positive for CD9, CD13, CD22, CD25 (dim), CD33, CD36, CD38 (bright), CD45 (dimmer than lymphocytes and brighter than myeloblasts), and CD123 (bright), and were negative for CD19, CD34, CD64, CD117, and HLA-DR. Basophils in all chronic myelogenous leukemia patients possessed 1 to 5 immunophenotypic aberrancies. The most common aberrancies were underexpression of CD38, followed by aberrant expression of CD64 and underexpression of CD123. CD34 and CD117 were present in cases with basophilic precursors. Myeloblasts showed a distinct immunophenotypic profile, as they typically expressed CD34 and CD117, showed dimmer expression (compared with basophils) of CD38, CD45, and CD123, and lacked expression of CD22. Conclusions.—Flow cytometric immunophenotyping can identify immunophenotypic aberrations of basophils in chronic myelogenous leukemia, and discriminate basophils from myeloblasts.

scholarly journals Clinical and Morphological Profile of Immune Thrombocytopenic Purpura in Children - A Five Year Study in a Paediatric Tertiary Health Care Centre of South India

2020 ◽  
Vol 7 (46) ◽  
pp. 2724-2729
Author(s):  
Ashida M. Krishnan ◽  
Deepthi Raj M.L ◽  
Priya V.S ◽  
Arya R.S
Keyword(s):  
Health Care ◽  
Bone Marrow ◽  
Immune Thrombocytopenic Purpura ◽  
Peripheral Blood ◽  
South India ◽  
P Value ◽  
Severe Thrombocytopenia ◽  
Care Centre ◽  
Health Care Centre ◽  
Thrombocytopenic Purpura

BACKGROUND Immune Thrombocytopenic Purpura (ITP) is one of the most commonly encountered disease in paediatric practice. Thorough clinical and morphological study of peripheral blood and bone marrow is required for confirming ITP. Clinicomorphological aspects of paediatric ITP is a less studied topic especially in developing countries like India. The objective was to study the clinical and morphological profile of paediatric cases of ITP. METHODS This is a 5-year record based retrospective study conducted in a paediatric tertiary health care centre in Kerala, South India. Data of all paediatric cases diagnosed as ITP including clinical presentation, clinical findings, blood counts, peripheral blood morphology, bone marrow morphology, and treatment response was collected and entered in SPSS software version 16.0 and analysed. For assessing correlation, chi-square test was used. RESULTS The age of children ranged from 3 months to 15 years. H/o viral fever was noted in 53 % cases. Cases which had moderate and severe thrombocytopenia were 74 % and 21 % respectively. Isolated thrombocytopenia was the most common peripheral blood picture observed with few cases showing coexisting eosinophilia and anaemia. All cases showed megakaryocyte proliferation in marrow with 9 % cases showing coexisting iron deficiency anaemia. Majority of cases showed rapid response to steroid / IVIG therapy and the response had no correlation with grade of thrombocytopenia (p value < 0.05). CONCLUSIONS Paediatric cases of ITP usually present following viral infections or vaccination, with worrisome bleeding episodes, petechiae, ecchymosis or purpura. KEYWORDS ITP, Paediatrics, Platelet Count, Thrombocytopenia, Vaccination

scholarly journals Bone Marrow and Peripheral Blood Cells Toxicity of a Single 2.0 Gy Cobalt60 Ionizing Radiation: An Animal Model

1970 ◽  
Vol 29 (2) ◽  
Author(s):  
Shittu Akeem ◽  
Olatunbosun Lukman ◽  
Khalil Eltahir ◽  
Olalere Fatai ◽  
Babatunde Abiola ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Peripheral Blood ◽  
Blood Cells ◽  
Marrow Cell ◽  
P Value ◽  
Peripheral Blood Cells ◽  
P Values ◽  
Cell Parameters ◽  
Post Irradiation ◽  
Total Body

BACKGROUND: Bone marrow is extremely vulnerable to damage caused by radiation therapy. Hence, bone marrow suppression is an important side effect of radiotherapy. Effective use of radiotherapy is therefore compromised by radiation-related injuries.MATERIAL AND METHODS: Six Guinea-pigs were recruited for the study of which three were subjected to total body irradiation with Co60 while the other three served as controls. Bone marrow and peripheral blood samples were collected before and at days 9, 14 and 21, post irradiation. Manual and automated counts were performed for bone marrow nucleated cells and peripheral blood cells respectively.RESULTS: Declining bone marrow cellularity was evident immediately post irradiation. Mean ± SD of marrow cell counted per mm3 were 121,924±281, 87,603±772, 121,367±375 and122,750±1000 pre-irradiation and days 9, 14 and 21, postirradiation (p-values 0.10, 0.27 and 0.29 respectively). Significant drops in counts were noticed on day 9 post-irradiation for all red cell parameters (p-values <0.05), for Total White Blood Cell Count and Neutrophil count (p-values <0.05) and also on days 14 and 21 for Lymphocytes (p-values <0.05) and on day 21 for Eosinophil/Basophil/Monocytes (p-value <0.05). A significant drop in platelets counts was also noticed on day 9 (p-value <0.05) which significantly increased above pre-irradiation value on day 21.CONCLUSION: Total body irrradiation with Co60 significantly affects the bone marrow with maximum reductions in marrow nucleated cells and peripheral blood cells counts on day 9 post irradiation. 

scholarly journals SMOKING GENOTOXICITY IN PERIPHERAL BLOOD LYMPHOCYTES USING THE COMET ASSAY

2013 ◽  
Vol 03 (03) ◽  
pp. 038-041
Author(s):  
Shobha S. Shetty ◽  
Hrishikesh Nachane
Keyword(s):  
Dna Damage ◽  
Comet Assay ◽  
Peripheral Blood ◽  
Peripheral Blood Lymphocytes ◽  
The Body ◽  
P Value ◽  
Tail Moment ◽  
Blood Lymphocytes ◽  
Significant Difference ◽  
Almost All

Abstract Background: Smoking has been shown to have a positive effect on DNA damage in almost all the cells of the body. Quantitative analysis of this damage will help in assessing the etiopathogenesis of various nicotine induced damage to the body. Comet assay has been an emerging tool in this regard and hence was applied by us to estimate the severity of DNA damage in smokers. Aims & Objectives: To evaluate the DNA genotoxicity in peripheral blood lymphocytes in smokers and their comparison with non smokers & assess the quantitative damage. Materials and methods: 30 smokers & 20 non smokers were recruited & their peripheral blood was taken for the comet assay to look for Olive moment & Tail moment to quantitatively assess the DNA damage due to cigarette smoking. Results: In our study there was no significant difference in the analysis of DNA damage (with regard to tail moment & olive moment) in smokers versus non smokers (P value: more than 0.05). Conclusions: Though smoking is known to cause DNA damage, we did not find significant differences between the two groups probably due to other multifactorial etiologies for genotoxicity.

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