Effects of thymulin 5cH in granuloma evolution and B1 cell differentiation: an experimental model to understand its biological mechanisms

2021 ◽  
Vol 10 (36) ◽  
pp. 194-195
Author(s):  
Cesar Sato ◽  
Thayná Neves Cardoso ◽  
Lika Osugui ◽  
Ana Flavia Popi ◽  
Leoni Villano Bonamin
Keyword(s):  
Flow Cytometry ◽  
Cell Differentiation ◽  
Experimental Model ◽  
Broiler Chickens ◽  
Blue Staining ◽  
Thymic Hormone ◽  
Macrophage Activity ◽  
Group A ◽  
Staining Methods ◽  
Group B

In previous studies, we found that thymulin (a thymic hormone), when prepared in homeopathic 5cH potency, had the property to improve the productive performance of broiler chickens infected with reovirus, as well as modulate the development of Ehrlich tumor and granuloma inflammatory lesions in mice by immune-mediated mechanisms. The aim of the present work was to study the immunomodulatory mechanisms of thymulin 5cH in a granuloma experimental model, by subcutaneous inoculation of BCG in mice, focusing the B-1 cells and zinc involvement in this process. Three groups of male Balb/c SPF mice (group A treated with thymulin 5cH, group B treated with thymulin 5cH incubated in Chelex ® - a zinc chelant - and group C, control, treated with vehicle) were inoculated with BCG in the left footpad and subcutaneous granuloma and spleen were harvested for histomorphometry analysis, after 7, 14 and 21 days. Ziehl-Neelsen, HE and Prussia Blue staining methods were used. Flow cytometry was also used in the same times to characterize and quantify peritoneal cells. Positive cells for CD11b (activated phagocytes, B-1 cells), CD19 (B-1 and B-2 cells), CD23 (negative B-1 cells, positive B2 cells) and CD5 (B-1a cells) were analyzed in a FACS Calibur (BD) device. Statistical analysis was performed using Kruskal - Wallis / Dunn for nonparametric evaluations and ANOVA / Tuckey-Krammer for the parametric ones. The X² method was used to evaluate the cell count in flow cytometry. P values ≤ 0.05 were considered statistically significant. Mice treated with thymulin 5cH presented higher macrophage activity and increase in the follicular area were seen in spleen after 7 days. Increase in gross lesion diameter and decrease in local BCG infection were seen after 21 days. At this time, the flow cytometry demonstrated the increase in peritoneal phagocytes derived from B-1 cells in thymulin 5cH treated mice, independently of Chelex ® incubation. The incubation of thymulin 5cH with Chelex ® blocked its effects only upon the number of B2 cells in the peritoneum and reduces Mn levels in the medicine solution. We conclude that thymulin 5CH modulates the BCG-induced granuloma through more than one mechanism, especially by peritoneal B1 cell differentiation into phagocytes.

scholarly journals Chest Mobility Exercise with Staked Breathing Versus Chest Mobility Exercises with Incentive Spirometery On Chest Expansion with Pleural Effusion Patient: A Comparative Study

2021 ◽  
Vol 9 (4) ◽  
pp. 3949-3953
Author(s):  
Minhaj Tahir ◽  
◽  
Tahzeeb Fatima ◽  
Devendra Trivedi ◽  
Manjit Kumar ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Pleural Effusion ◽  
P Value ◽  
Chest Expansion ◽  
Before And After ◽  
Group A ◽  
Group B ◽  
Pleura Effusion ◽  
Respiratory Conditions ◽  
Incentive Spirometry

Background: Pleural effusion is one of the commonly seen respiratory conditions in India with approximately 1 million people being diagnosed each year. Twenty to forty percent of hospitalized patients with bacterial pneumonia develop pleural effusion. In India unlike western countries, tuberculosis pleura effusion is common. The pleural cavity is involved in approximately 5% of all patients with tuberculosis. Since there was no literature regarding the effectiveness chest mobility exercise with staked breathing or chest mobility exercises with incentive spirometery in pleural effusion. There was a need to find out as to which approach are the best ones to implement. Objective: To compare the efficacy of chest mobility exercise with stacked breathing versus chest mobility exercise with incentive spirometery on chest expansion in patients with pleural effusion. Materials and Method: 20 patients with pleural effusion were selected by easy sampling and randomly assigned into two groups (10 patients each groups). Group A received chest mobility exercises and intensive spirometery and group B received chest mobility exercises and stacked breathing. Both groups were instructed to perform the intervention 3 time per day, 8 to 10 time per session for one week. Chest expansion was measured by thoracic flow cytometry before and after one week of intervention. Result: In group A chest expansion increase from 2.68 to 2.87 which was statistically significant (P value < 0.0023). In Group B the chest expansion increases from 2.94 to 3.09 which was not statistically significant (P value < 0.216). Conclusion: It was concluded from the result that both chest mobility exercises with intensive spirometery and chest mobility exercise with stacked breathing are equally effective in improving the chest expansion in subject with pleural effusion. KEY WORDS: Pleural effusion, Chest mobility exercises, Incentive Spirometry, Stacked breathing, Thoracic flow cytometry.

scholarly journals A substitute to xylene in deparaffinization and clearing prior to coverslipping in histopathology

2019 ◽  
Vol 11 (02) ◽  
pp. 118-122
Author(s):  
Nasar Yousuf Alwahaibi ◽  
Sirin Hamed Aldughaishi
Keyword(s):  
Cost Benefit ◽  
Cost Effective ◽  
Nuclear Staining ◽  
Laboratory Cost ◽  
Cytoplasmic Staining ◽  
Group A ◽  
Staining Methods ◽  
Laboratory Workers ◽  
Group B ◽  
Group D

Abstract INTRODUCTION: Deparaffinization and clearing prior to coverslipping are important steps in all staining methods in histopathology. Xylene is the most commonly used agent worldwide. However, xylene is toxic. We evaluated safer alternative dewaxing and clearing agents prior to coverslipping in a histopathology laboratory. MATERIALS AND METHODS: Thirteen different fresh surgical tissues were cut into two halves. One half processed using xylene and the other half processed using UltraClear™. Five groups were designed. For each Group of A, B, C, and D, 100 slides were cut from xylene-processed blocks. For Group E, 100 slides were cut from UltraClear™-processed blocks. Group A is the standard method. Group B evaluates UltraClear™ as a dewaxing agent only. Group C evaluates UltraClear™ as a clearing agent prior to coverslipping only. Group D evaluates UltraClear™ as both dewaxing and clearing agents prior to coverslipping. Group E evaluates UltraClear™ as both dewaxing and clearing agents prior to coverslipping. Six parameters were evaluated: nuclear staining, cytoplasmic staining, cell morphology, clarity of staining, uniformity of staining, and cost. RESULTS: Groups B, C, and D showed 79% (P = 0.054), 83% (P = 0.221), and 80% (P = 0.079) adequacy when compared with Group A (89%), respectively. However, Group E showed only 76% (P = 0.016) adequacy. UltraClear™ is more expensive than xylene. CONCLUSION: UltraClear™ is a promising dewaxing agent. It is also a good clearing agent for use prior to coverslipping in histopathology laboratory. Cost-benefit balance between safety of laboratory workers, good quality staining, and cost-effective strategy needs to be further studied.

scholarly journals Laser Doppler Flow Imaging of Open Lower Leg Fractures in an Animal Experimental Model

2002 ◽  
Vol 10 (2) ◽  
pp. 114-119 ◽  
Author(s):  
L Herzog ◽  
FX Huber ◽  
PJ Meeder ◽  
G Muhr ◽  
J Buchholz
Keyword(s):  
Experimental Model ◽  
Laser Doppler Flowmetry ◽  
Tibial Fracture ◽  
Muscle Flap ◽  
Lower Leg ◽  
Laser Doppler ◽  
Doppler Flowmetry ◽  
Group A ◽  
Group B ◽  
Animal Experimental Model

Purpose. Open lower leg fractures are frequently associated with severe soft tissue damage, followed by osteomyelitis. Using an animal experimental model, we investigated the effect of timing of coverage of a tibial fracture with a local muscle flap. Methods. 80 rabbits had a tibial fracture induced in a standardised fashion, which was stabilised by screw osteosynthesis. After 3 (group A; n=40) and 7 days (group B; n=40), respectively, the tissue defect was covered by a local gastrocnemius flap. In increasing intervals from 1 to 2, 4, 8, and 16 weeks, the rabbits from each group were killed and the bone fracture was analysed histomorphologically Cortical microcirculation was measured by 2-channel laser doppler flowmetry. Results. Muscle flaps after 3 days improved perfusion significantly as compared with 7 days (24 Flux [standard error, 5 Flux] versus 10 Flux [3 Flux]; baseline, 1.4 Flux). Group A animals also displayed a lower rate of necrosis (0 versus 38). The incidence of osteomyelitis was higher in group B than in group A (24% versus 0%). Conclusion. Laser doppler flowmetry was proven to be a reliable, minimally invasive means for identifying avital tissue, leading to reduction in the loss of vital bone tissue in experimental settings.

Concurrent Treatment with Denileukin Diftitox (DD) To Deplete T-Regulatory Cells Enhances Rebound Lymphocytosis and Eosinophilia in Patients Treated with High-Dose IL-2 (HDIL-2) for Metastatic Renal Cell Cancer (MRCC).

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 1729-1729
Author(s):  
Adi Gidron ◽  
John Eklund ◽  
Brenda Martone ◽  
Alfred W. Rademaker ◽  
Charles Goolsby ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Immune Response ◽  
Statistical Significance ◽  
Cell Cancer ◽  
Peak Level ◽  
High Dose ◽  
Maximal Increase ◽  
Group A ◽  
Group B ◽  
Group D

Abstract Background: CD4+CD25+hi T cells (Treg) play a suppressive role in immune regulation. DD is an IL-2 receptor specific cytotoxin. We postulated depletion of Treg with DD may enhance immune effector cell populations after HDIL-2 treatment, including rebound lymphocytosis and also eosinophilia which has been reported to be involved in immune response to neoplasm (Mattes J Exp Med 197: 387, 2003). Methods: In this pilot study, 12 pts (8 male, median age 58 yrs) with MRCC were tx with HDIL-2 and DD in different schedules to determine safety and effect on immune response as manifested by changes in Treg, peak lymphocyte, and eosinophil counts. Pts were treated with IL-2 600,000 IU/kg Q8H on days (d) 1–5 and 15–19. Three (group A) and 4 (group B) pts were given 6 and 9ug/kg daily on d8–10 respectively, while 5 (group C) pts received 9ug/kg of DD on d −4 to −2. Nine (group D) pts with metastatic melanoma who received HDIL-2 as above but without DD were included as controls. Flow cytometry was done on days −4, 1,8,10,15,22 for group C and on days 1,8,10,15,22 for groups B, and D. CBC was obtained concurrent or within 24 hours of flow cytometry. Group A pts were evaluated for safety only and were excluded from analysis. Results: Prior to enrollment, all pts had undergone nephrectomy and four patients received interferon-alpha. One pt from group B withdrew from study and was not included in analysis. Administration of DD resulted in a median decline of 25% in Treg number (not significant). DD given before HDIL-2 was associated with a greater increase in Treg post HDIL-2. In Group C there was an increase of rebound median Treg count of 0.88k/ul compared with 0.060k/ul in group B (p=0.025). Absolute lymphocytosis was higher in the combined group getting DD compared to control (median maximal increase of 7.6 vs 4.7 k/ul, respectively) although the difference did not reach statistical significance. However, group C pts had a greater increase in absolute lymphocytosis than did group B pts in which absolute lymphocytosis actually decreased (median increase 10.6 vs. median decrease 0.4 k/ul, p=0.025). A higher peak level of eosinophilia was noted in groups B and C compared with group D (mean increase of 10.5 vs. 4.0 k/ul p=0.2). Group C had a greater peak eosinophilia than group B (11.2 vs 2.2 k/ul p=0.053) Toxicity was manageable and consistent with those seen with HDIL-2. Median HDIL-2 dose given was 21 (range, 14–28). No clinical responses were observed. Of 11 pts included in the analysis 1 pt from group A expired 68 weeks after enrollment. All remaining patients are alive. Survival from enrollment ranges from 11 to 93 weeks. Conclusion: Overall, the combination of DD and HDIL-2 results in a stimulatory effect as manifested by increased rebound lymphocytosis and eosinophilia compared to HDIL-2 alone. Administration of DD in conjunction with HDIL-2 was associated with a rebound in Treg that may be schedule and dose dependent. The results suggest an enhanced immune stimulatory effect as manifested by lymphocytosis and peak eosinophilia in group C. However, this stimulatory effect also extends to Treg that may prove detrimental clinically. Further exploration of these effects in immunotherapy naïve patients would be beneficial.

Microrna Expression Profile Identifies Distinct Clinically Relevant Sub-Groups in Multiple Myeloma: Novel Prognostic Markers and Potential Targets for Therapy

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 96-96 ◽  
Author(s):  
Sophia Adamia ◽  
Herve AvetLoiseau ◽  
Samirkumar B Amin ◽  
Yu-Tzu Tai ◽  
Steven P. Treon ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Cell Differentiation ◽  
Clinical Outcome ◽  
Signaling Pathways ◽  
Cell Lines ◽  
Plasma Cells ◽  
Normal Plasma ◽  
Group A ◽  
Group B

Abstract MicroRNA, an abundant class of small endogenous RNAs, regulate target genes through inducing translational inhibition and cleavage of targeted transcripts. To date, microRNAs have been implicated in normal biological processes, including development, cell differentiation, apoptosis and proliferation as well as in malignant transformation. However, their role in multiple myeloma (MM) remains unknown. Here we investigated role of microRNAs in myelomagenesis, and their influence on prognosis and clinical outcome. We evaluated profiles of 384 microRNAs in bone marrow derived CD138+ plasma cells (PC) from 79 uniformly treated MM patients, 11 MM cell lines and 9 healthy donors using qRT-PCR based microRNA array. The relative expression was calculated using comparative Ct method, and data was normalized using endogenous controls and analyzed using SDS, RQ manager, R and dChip softwares. MicroRNA expression profiles detected in MM patients were correlated with clinical outcome measures. We observed significant modulate expression of 61 microRNAs in myeloma cells compared to normal plasma cells. When more stringent criteria were used, we identified 24 differentially expressed microRNAs in patient samples. Further, unsupervised hierarchical clustering of filtered microRNAs, based on their DCt values, identified two major groups within the MM population (groups A and group B). Samples of Group A clusters with MM cell lines, indicating more proliferative nature of MM patient cells. Within B group, a second degree node group B2, clusters with normal plasma cells indicating more indolent course, while patients in an additional node B1 represented an assorted pattern. The unsupervised clustering of all MM samples showed consistent changes in miR-30b, -30c, -30d, -142-5p, -24, -191, -181d, -374, -146b, -140, -145, -125a, -151, -223, -155, let7b, indicative of a role of these microRNA in myelomagenesis; while supervised analysis of samples within groups A and B identified modulated expression of different sets of miRNAs. In group A miR-585 and let-7f were upregulated 8–12 fold, while miRs -125a, -126, -155, -223, -146a, -374 -19a, -20a, -26a, -30a -5p, -30b, and -30d were significantly downregulated; in group B, all differentially expressed microRNAs were downregulated (p<0.001) compared to normal plasma cells. These modulated miRNAs target critical signaling pathways including apoptosis, hematopoietic cell differentiation and proliferation, survival and angiogenesis by upregulating function of HOX9, c-myc, VCAM-1, Bcl-2, E2F1, SHP1, SHP2, VEGF, and DUSp6 molecules. We further analyzed the effect of microRNA on clinical outcome. We have observed significantly superior event free and overall survival of patients in group B2 compared to patients in group A (2 yr estimated EFS 79% versus 54% respectively; p=0.05; and 2 yr estimated OS 94% versus 70% respectively; p =0.017). Taken together this data identifies critical microRNAs as modulators of gene expression and signaling pathways and provides potential novel microRNA and gene targets in MM to both understand biological behavior and for therapeutic application.

scholarly journals Experimental model of the morphological and morphometric aspects of tissue repair in skin wounds submitted to beta radiation emitted by Strontium -90 in rats

2004 ◽  
Vol 19 (suppl 1) ◽  
pp. 51-58
Author(s):  
Gerson Vilhena Pereira Filho ◽  
José Mario Camelo-Nunes ◽  
Fabrício Yui
Keyword(s):  
Experimental Model ◽  
Optical Microscope ◽  
Beta Radiation ◽  
Daily Group ◽  
Morphometrical Analysis ◽  
Strontium 90 ◽  
Group A ◽  
Group B ◽  
And Control ◽  
Made In

The purperose of this work is tell the use of the experimental model to investigation of the effects of Beta radiation of estroncium-90 for repairing the tissue of wounds made in rats. 48 animals of the lineage EPM-1 Wistar were used, distributed in two groups that received radiation in alternate days (group A) and daily (group B). Each group was divided in four groups of six rats to be analysed in the 3rd, 7th, 14th and 21st day after operation. Two incisions were made in the back of the animal and sutured immediately afterwards; the upper part was irradiated and the lower part used as control. On the dates established the irradiated and control wounds were macroscopically examined and withdrawn for preparation of the histological comparative study in the optical microscope. Following, an morphometrical analysis was performed to count leucocyts, fiberblast and colagen fibers which were submitted to statistical study.

scholarly journals Physicochemical Assessment of Broiler Chickens Fed Diets Supplemented with a Mixture of Ginger, Garlic and Cinnamon

2020 ◽  
Vol 24 (5) ◽  
pp. 809-814
Author(s):  
A.S. Idoko ◽  
A. Zaharaddeen ◽  
N.U. Imam ◽  
S. Nura ◽  
B. Abdulazeez ◽  
...  
Keyword(s):  
Weight Gain ◽  
Broiler Chickens ◽  
Carcass Characteristics ◽  
Broiler Chicks ◽  
Physicochemical Changes ◽  
Cholesterol Levels ◽  
Phase Group ◽  
Group A ◽  
Group B ◽  
Haematological Indices

Some physicochemical changes in broiler chickens fed diets supplemented with a mixture of ginger, garlic and cinnamon was evaluated. During starter phase, 150 broiler chicks were divided into groups A and B of 75 birds each, and were randomly fed diets 1 and 2 respectively for 3 weeks. The average weekly weight gain by the chickens maintained on the mixed spices supplemented diet (235.15±15.55g) was significantly lower (p<0.05)compared with the weight gain by the control (274.26±20.23g) in the Starter phase. In the Finisher phase, group A was sub-divided into groups C and D while group B was subdivided into groups E and F and fed for additional 3 weeks. The average weekly weight gain by groups C (242.31±4.97g), D (282.46±15.04g), E (260.14±81.11g) and F (236.67±24.29g) did not vary significantly. The evaluated carcass characteristics and haematological indices when compared with the control did not vary significantly. The serum cholesterol levels (mg/dl) in groups D (101.40±4.13), E (106.60±4.59) and F (100.20±9.83) were significantly lower (p<0.05) in comparison with the level in group C (127.00±8.83). Groups E (46.96±8.62mg/dl) and F (47.44±6.35mg/dl) had significantly (p<0.05) lower LDL-C compared with the control (70.56±8.75mg/dl). Therefore, mixture of ginger, garlic and cinnamon may have no effects on the overall weight gain, carcass traits and haematological profiles but could decrease the risk of atherogenesis and CVD in broiler chickens. Keywords: Cinnamon; ginger; garlic; spices; physicochemical

scholarly journals Immunophenotype Drift of Residual Plasma Cells Indicates Therapeutic Response and Prognosis in Multiple Myeloma

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 5491-5491
Author(s):  
Chenxing Du ◽  
An Gang ◽  
Yan Xu ◽  
Xuehan Mao ◽  
Yuting Yan ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Flow Cytometry ◽  
Therapeutic Response ◽  
Conflicts Of Interest ◽  
Early Stage ◽  
Therapy Group ◽  
Newly Diagnosed ◽  
Normal Phenotype ◽  
Group A ◽  
Group B

Background Chemotherapy resistance remains a significant hurdle in the treatment of multiple myeloma (MM). However, it is difficult to discriminate the potential refractory patients from the very early stage. Flow cytometry is a convenient tool to detect the residual myeloma cell tiding, indicating therapeutic response sensitively. Methods From June, 2014 to December, 2016, 172 sequential patients with newly diagnosed multiple myeloma were enrolled in the BDH2008/02 clinical trial. Patient informed consent was obtained in accordance with the Declaration of Helsinki. 144 patients with at least two flow cytometry detections were analyzed. Bone marrow samples were detected by an eight-color EuroFlow panel. CD20 negative and CD81 positive is defined as normal phenotype. Results We conducted a median of 3-time (2-8) flow cytometry detection on each patient. When newly diagnosed and achieved best response, CD20, CD81 expression rates were 29.9%, 9.7% and 14.9%, 64.4% (P=0.0091, P<0.0001), respectively. According to the status variation of CD20 and CD81, all patients were divided into three groups: both markers were always normal (Group A), either CD20 or CD81 was abnormal at diagnosed and turned normal during therapy (Group B) and markers stayed abnormal (Group C). Patients with undetectable residual tumor cells were also classified as Group A. The overall response rate of the patients in Group C was inferior to Group B (>PR rate: 54.3% vs. 71.4%, P=0.021). And the OS of Group C was significantly worse than Group A and B (47.9 months vs. not reached vs. not reached, P=0.036). Conclusion CD20/CD81 switching to normal phenotype during therapy indicates therapeutic response and an improved outcome than that staying abnormal. The expression tiding of CD20 and CD81 may be a reasonable combination to dynamically stratify MM patients, directing the choice of maintenance therapy. Figure Disclosures No relevant conflicts of interest to declare.

scholarly journals Antiarrhythmic Effect of Ranolazine in Combination with Selective NCX-Inhibition in an Experimental Model of Atrial Fibrillation

2020 ◽  
Vol 13 (10) ◽  
pp. 321
Author(s):  
Julian Wolfes ◽  
Christian Ellermann ◽  
Niklas Broer ◽  
Benjamin Rath ◽  
Kevin Willy ◽  
...  
Keyword(s):  
Atrial Fibrillation ◽  
Experimental Model ◽  
Antiarrhythmic Effect ◽  
Refractory Periods ◽  
New Zealand White Rabbits ◽  
Monophasic Action Potentials ◽  
Cycle Lengths ◽  
Group A ◽  
Group B ◽  
Whole Heart

The aim of this study was to investigate the effects of a combination of ranolazine with different selective inhibitors of the Na+/Ca2+-exchanger (NCX) in an established experimental model of atrial fibrillation (AF). Eighteen hearts of New Zealand white rabbits were retrogradely perfused. Atrial catheters were used to record monophasic action potentials (aPRR). Hearts were paced at three different cycle lengths. Thereby, atrial action potential durations (aAPD90), atrial effective refractory periods (aERP) and atrial post-repolarization refractoriness were obtained. Isoproterenol and acetylcholine were employed to increase the occurrence of AF. Thereafter, the hearts were assigned to two groups (n = 9 each group) and additionally perfused with a combination of 10 µM ranolazine and 1 µM of the selective NCX-inhibitor ORM-10103 (group A: Rano-ORM) or 10 µM ranolazine and 1 µM of another NCX-inhibitor, SEA0400 (group B: Rano-SEA). The infusion of Iso/ACh led to a shortening of aAPD90, aERP, aPRR and the occurrence of AF episodes was significantly increased. Additional perfusion with ranolazine and ORM-10103 (group A) significantly prolonged the refractory periods and aPRR and AF episodes were effectively reduced. In group B, Rano-SEA led to a slight decrease in aAPD90 while aERP and aPRR were prolonged. The occurrence of AF episodes was consecutively reduced. To our knowledge, this is the first study investigating the effect of ranolazine combined with different selective NCX-inhibitors in an isolated whole-heart model of AF. Both combinations prolonged aERP and aPRR and thereby suppressed the induction of AF.

A Useful Flow Cytometry Panel To Exclude Myelodysplastic Syndrome.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 4848-4848
Author(s):  
Ling Zhang ◽  
Jean R. Lopatequi ◽  
Sharron E. Kelly ◽  
Tobi Neer ◽  
Stephen Lee
Keyword(s):  
Bone Marrow ◽  
Flow Cytometry ◽  
Test Group ◽  
Bone Marrow Examination ◽  
Normal Group ◽  
P Value ◽  
Aberrant Expression ◽  
Cd10 Expression ◽  
Group A ◽  
Group B

Abstract Background: Myelodysplastic syndromes (MDS) are a heterogeneous group of hematopoietic disorders, which can be difficult to diagnose based only on morphologic bone marrow examination. Karyotyping can be useful in diagnosing borderline cases. However, only 40% of patients with MDS have an abnormal karyotype. Flow cytometry(FCM) approaches have been described but not clearly defined in MDS due to use of different criteria. We devised a 10-parameter FCM panel, mainly including myeloid and erythroid maturation markers, to differentiate MDS marrows from normal marrows. Design: Bone marrow from 91 patients with cytopenia(s) or anemia were included in the study(10/2005–7/2006). Cases were divided into 3 groups: normal, not morphologically suspicious for MDS, 46 cases, equivocal, morphologically suggestive but not diagnostic of MDS, 20 cases, morphologically diagnostic of MDS, 25 cases. 10 FCM paremeters were performed and scored: hypogranularity,aberrant expression of CD56,lack of CD10 expression,decreased CD64 expression,&lack of CD13 or CD33 expression,&abnormal CD13/CD16 or CD16/CD11b pattern,increase of CD 34 expression gating on all cells excluding erythrocytes,decreased expression of CD71/Glycophorin gating on erythroid precursors. Karyotypings were analyzed. Results: and (see Table 1 and 2). Karyotyping were anlayzed in 86 cases. Cytogenetic abnormalities were found in 2.2% (1/44) of normal group, 5.3% (1/19) of equivocal group and 34.8% (8/43) of MDS group. In MDS group 7 of 8 patients (87.5%) who had both morphologic and cytogenetic diagnosis of MDS were scored &gt;=3 of 8 FCM parameters. Conclusions: Study showed the 8-parameter FCM panel was more predictive of MDS than 10-parameters one. Both CD13/CD16 & CD16/CD11b patterns were considered to be non-specific. In 8-parameter panel, zero score tended to rule out MDS, while score &gt;=3 suggested MDS. The most specific FCM parameters were hypogranularity, CD34, aberrant expression of CD56 or lack of CD10 expression by mature granulocytes. CD71/Glycophorin A might be useful in identifying dysplasia in erythroid lineage. Table 1. Comparison of FCM Parameters in Patients with/without MDS Parameters/Group Normal Group(A)(n=46) Equivocal Group(B)(n=20) MDS Group(C)(n=25) CHITEST(P value)(GRoup A vs C) * These two parameters were deleted in the 8-parameter panel due to their high frequency seen in normal group. Hypogranularity 4 (8.7%) 6 (30%) 18 (72%) 0.0001 CD56 0 (0.0%) 1 (5%) 6 (24%) 0.0005 CD10 7 (15.2%) 7 (35%) 15 (60%) 0.0001 CD64 6 (13.1%) 2 (10%) 6 (24%) 0.2396 CD13 0 (0.0%) 0 (0.0%) 2 (8.0%) 0.0516 CD33 0 (0.0%) 2 (10 %) 0 (0%) 0 CD13/CD16* 23 (50.0%) 7 (35%) 21 (84%) 0.0047 CD34 8 (17.4%) 7 (35%) 13 (52%) 0.0026 CD71 2 (4.3%) 5 (25%) 12 (48%) 0.0001 CD16/CD11b* 27 (58.7%) 8 (40%) 22 (88%) 0.0107 Table 2. Scoring with 8 FCM Parameters in Patients with/without MDS Score/Group Normal Group (A)* (n=46) Equivocal Group (B)* (n=20) MDS Group (C)* (n=25) * Fish Exact T Test: Group A vs C: p&lt;0.0001, A vs B: p=0.0006, B vs C: p=&lt;0.0001 Score=0 21 (45.7%) 3 (15%) 0 (0.0%) Score=1–2 25 (543 %) 14 (70%) 8 (32 %) Score&gt;3 0 (0.0%) 3 (15%) 17 (68%) *Mean score +/− SD 0.65+/−1.06 1.55+/−1.54 2.8+/−1.65

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