Hemopoietic Disorders Research Articles

Analyzing the characteristics and prognostic outcomes for patients with myelodysplastic syndrome with Q-gene deletion.

e18526 Background: Myelodysplastic syndromes (MDS) are a group of hemopoietic disorders in which the bone marrow is deficient in producing healthy mature blood cells. MDS associated with a deletion in the long arm of chromosome 5Q is a rare occurrence, but considered a good prognosis in comparison to other subgroups of MDS. There has been limited focus directed towards this variant of the disease, especially concerning outcomes among different demographics. Methods: The National Cancer Institute’s Surveillance, Epidemiology, and End-Result (SEER) registry research database 17 (2000 -2020) was used to assess & compare the outcomes for this variant of MDS in adult & AYA pts. Univariate & multivariate analyses was performed and cox proportional hazards regression was used to assess prognostic outcomes. A two-sided pvalue ≤0.05 was considered significant. Results: Analysing (table) 2,323 pts identified, 98.92% (n= 2298) were adults ≥40yrs, and 1.08% (n= 25) were AYAs. AYAs comprised 32% Hispanic, 4% Non-Hispanic Asian or Pacific Islander (NHAPI), 16% Non-Hispanic Black (NHB), and 48% Non-Hispanic White (NHW). Compared to adults, AYAs had higher proportions of Hispanics and NHBs but fewer NHWs. AYAs showed longer average survival (57.7 vs 33 months) than older adults. Females generally had longer survival periods than males in both AYA (64.4 vs 49.1 months) & adult (37.2 vs 26.4 months) groups. Among adult racial groups, Non-Hispanic Black females had the poorest prognosis (29.6 months), while Hispanic males had the worst prognosis overall (21 months). Among AYAs, Hispanic females had the worst prognosis (4 months), followed by Hispanic males (24.2 months). Conclusions: Our study shows AYAs are less likely to die from MDS compared to adults ≥40. Hispanic females appear to have a worse outcome than males in both age groups & is independent of other analysed prognostic factors. More research is needed to examine other prognostic factors with the aim of improving survival outcomes. [Table: see text]

Upregulation of Platelet Transcripts As Biomarkers of Fibrotic Progression in Myeloproliferative Neoplasms

Myeloproliferative neoplasms (MPN) are a group of clonal hemopoietic disorders that have a proliferative phase (including essential thrombocythemia (ET) and polycythemia vera (PV)), and which can progress to advanced disease such as myelofibrosis (MF) or acute leukemia. This is thought to be due in part to progressively dysfunctional megakaryocytes driving reticulin deposition in the bone marrow. In previous work, we demonstrated that the platelet transcriptome in MPN differs from normal, and the difference is most marked with increased reticulin (Guo et al., 2020). A putative 3-gene signature ( CCND1, H2AFX and CEP55) could discriminate between patients with and without increased reticulin fibrosis. To progress this, we have profiled the transcriptome of platelets from a further 173 MPN patients were analyzed using next generation sequencing. CCND1, H2AFX and CEP55 were upregulated in platelets from patients with MF compared to ET (4.6-fold, padj<0.001, 9.0-fold, padj<0.001 and 10.5-fold, padj<0.001, respectively) and to PV (3.9-fold, padj<0.001, 9.2-fold, padj<0.001, 12.0-fold, padj<0.001). Application of the established 3-gene signature had a positive predictive value of 78%, a negative prediction value of 90% and accuracy of 88% for MF. To assess the temporal changes in these gene transcripts in platelets and the potential utility to predict fibrotic progression, we then performed longitudinal analyses in two MPN mouse models. Jak2V617F knock-in mice ( n=25) with stable serially transplantable disease, which resembles PV (Mullally et al., 2010), were sacrificed at 4, 8, 12, 16 and 20 weeks post-transplant, and controls ( n=5) at 20 weeks. Blood counts were performed, and spleen and sternum harvested and stained with hematoxylin and eosin, and for reticulin. By 20-weeks, the platelet and leucocyte counts and hemoglobin had increased as expected. There was splenomegaly with extramedullary hematopoiesis and a mild increase in marrow reticulin. There was no significant up-regulation of Ccnd1, H2afx or Cep55 in platelet transcripts at any timepoint compared to the control mice. This data showed stability of the mouse PV phenotype and platelet transcripts, in keeping with the chronic phase of MPN. Mice with MPLW515L ( n=8) (Pikman et al., 2006) were sacrificed at 3 and 6 weeks post-transplant, and control mice ( n=4) at 6 weeks. At 3-weeks post-transplant Cep55 (131.6-fold, padj<0.001) and H2afx (1.7-fold, padj<0.01) had increased in the MPLW515L mice and there was no increase in reticulin in the bone marrow. At 6-weeks platelet and leucocyte counts had increased, there was splenomegaly with extramedullary hematopoiesis including marked megakaryocytosis and marrow reticulin. On platelet transcript analysis Cep55 (154.6-fold, padj<0.001) and H2afx (2.4-fold, padj<0.001) were significantly increased; there was no significant change for Ccnd1. Upregulation of Cep55 and H2afx in the MPLW515L mouse occurred prior to the accumulation of bone marrow reticulin and development of MF phenotype. This suggests that up-regulation of Cep55 and H2afx in platelets may be a predictor of disease progression from ET and PV to MF. For MPN patients, a deviation from steady-state platelet transcript levels, specifically for H2AFX and CEP55, may infer a biological transition towards the fibrotic phenotype. This novel approach utilizing platelet transcript analysis may therefore have potential for serially monitoring MPN patients and the early identification of genomic changes that may herald disease transformation to MF.

Antibodies to calnexin and mutated calreticulin are common in human sera

Purpose of the studyCalreticulin is an endoplasmic reticulum chaperone protein, which is involved in protein folding and in peptide loading of major histocompatibility complex class I molecules together with its homolog calnexin. Mutated calreticulin is associated with a group of hemopoietic disorders, especially myeloproliferative neoplasms. Currently only the cellular immune response to mutated calreticulin has been described, although preliminary findings have indicated that antibodies to mutated calreticulin are not specific for myeloproliferative disorders. These findings have prompted us to characterize the humoral immune response to mutated calreticulin and its chaperone homologue calnexin. Patients and methodsWe analyzed sera from myeloproliferative neoplasm patients, healthy donors and relapsing-remitting multiple sclerosis patients for the occurrence of autoantibodies to wild type and mutated calreticulin forms and to calnexin by enzyme-linked immunosorbent assay. ResultsAntibodies to mutated calreticulin and calnexin were present at similar levels in serum samples of myeloproliferative neoplasm and multiple sclerosis patients as well as healthy donors. Moreover, a high correlation between antibodies to mutated calreticulin and calnexin was seen for all patient and control groups. Epitope binding studies indicated that cross-reactive antibodies bound to a three-dimensional epitope encompassing a short linear sequence in the C-terminal of mutated calreticulin and calnexin. ConclusionCollectively, these findings indicate that calreticulin mutations may be common and not necessarily lead to onset of myeloproliferative neoplasm, possibly due to elimination of cells with mutations. This, in turn, may suggest that additional molecular changes may be required for development of myeloproliferative neoplasm.

Une alpha thalassémie acquise chez un patient de 86 ans avec un syndrome myélodysplasique

BackgroundAlpha thalassemia-myelodysplastic syndrome (ATMDS) is one of the possible complications related to the genetic instability typical of clonal hemopoietic disorders such as myelodysplastic syndromes (MDS). Hemoglobin H acquisition, which is hemoglobin without alpha chains and with 4 beta chains is the hallmark of this disease. ObservationAn 86-year-old male with chronic, microcytic anemia was referred due to a fall in his hemoglobin level. The blood smear was remarkable for intense anisocytoses and poikilocytosis. Bone marrow analysis was followed by a diagnosis of MDS with a good prognostic score. Peripheral blood coloration with brilliant cresyl blue showed “golf ball-like” erythrocytes. Hemoglobin electrophoresis is notable for the presence of H hemoglobin. The new generation sequencing confirmed the diagnosis of ATMDS showing a non-sense mutation in the gene ATRX. ConclusionThe diagnosis of ATMDS should be considered in the presence of the association of MDS, microcytic anemia and marked blood smear abnormalities such as anisocytosis and poikilocytosis. A little less than 10% of all MDS are complicated by ATMDS.

Study of bone marrow aspirations -5 years review from a single center in Kuwait- An audit

We reviewed the bone marrow examination (BME) studies performed in the last five-year period with the objective to find out the findings and ascertain the role of this procedure in the diagnosis at Al Adan hospital, this public teaching hospital of the state of Kuwait in Middle East. Samples were collected after history taking and clinical examination. Blood samples analyzed for CBC parameters along with biochemical and viral markers. Bone marrow aspiration samples were collected and stained with May Grunwald Giemsa stain, trephine biopsy processed and stained with Hematoxylin & Eosin along with Perl’s stain for iron and Reticulin stain. All samples were analyzed and results pooled by the team of hematopathologists. We reviewed 264 cases for BME and found ages ranging from 07 months to 95 years and 170 males and 94 females were included in the study. The array of conditions included 53% of clonal haemopoietic disorders including leukemias, Myelo Proliferative Neoplasm (MPN) , Myelo Dysplastic Syndrome (MDS), & Plasma cell disorders. 3.5% aplastic anemias, while only 5.4 % with nutritional causes and 28.5% of cases having various reactive causes; Metastatic deposits were found in 1.5% and haemophagocytosis in 3.5% of the subjects. Results were in concordance with studies conducted from developed nations. Our study concludes that bone marrow examination is crucial in diagnosis to elucidate the various etiology of afflictions of bone marrow and helps in proper management in this Middle East country.

The Hunt Is On! In Pursuit of the Ideal Stem Cell Population for Cartilage Regeneration.

Cartilage injury and degeneration are hallmarks of osteoarthritis (OA), the most common joint disease. OA is a major contributor to pain, loss of function, and reduced quality of life. Over the last decade, considerable research efforts have focused on cell-based therapies, including several stem cell-derived approaches to reverse the cartilage alterations associated with OA. Although several tissue sources for deriving cell-based therapies have been identified, none of the resident stem cell populations have adequately fulfilled the promise of curing OA. Indeed, many cell products do not contain true stem cells. As well, issues with aggressive marketing efforts, combined with a lack of evidence regarding efficacy, lead the several national regulatory bodies to discontinue the use of stem cell therapy for OA until more robust evidence becomes available. A review of the evidence is timely to address the status of cell-based cartilage regeneration. The promise of stem cell therapy is not new and has been used successfully to treat non-arthritic diseases, such as hematopoietic and muscle disorders. These fields of regenerative therapy have the advantage of a considerable foundation of knowledge in the area of stem cell repair mechanisms, the role of the stem cell niche, and niche-supporting cells. This foundation is lacking in the field of cartilage repair. So, where should we look for the ideal stem cell to regenerate cartilage? It has recently been discovered that cartilage itself may contain a population of SC-like progenitors. Other potential tissues include stem cell-rich dental pulp and the adolescent growth plate, the latter of which contains chondrocyte progenitors essential for producing the cartilage scaffold needed for bone growth. In this article, we review the progress on stem cell therapies for arthritic disorders, focusing on the various stem cell populations previously used for cartilage regeneration, successful cases of stem cell therapies in muscle and hemopoietic disorders, some of the reasons why these other fields have been successful (i.e., “lessons learned” to be applied to OA stem cell therapy), and finally, novel potential sources of stem cells for regenerating damaged cartilage in vivo.

Effect of Planting Season on the Residue of Organophosphate in Chili (Capsicum annuum L.)

Background: Pesticide residues harm human health and are associated with hemopoietic disorders and many organ cancers. This study analyzes the pesticide residue content in chili cultivated by farmers in rainy and dry seasons, especially for organophosphates pesticides. The residual difference in chili was affected by planting seasons caused by farmers’ adoption of crop cultivation technologies, including pesticide application. Methods: The analysis was carried out using gas chromatography on 26 samples from 13 chili cultivation locations in Aceh province, Indonesia. Sixteen samples were harvested during the rainy season and the remaining ten were harvested during the dry season. Result: The results showed that pesticide residues were found in two red chili samples planted in the rainy season, while no residue was found from the dry season. In the rainy season’s harvested samples, the Organophosphate pesticide residues were chlorpyrifos 0.52 mg kg-1 and 0.16 mg kg-1 at Mutiara Tiga sub-district and Meurebo sub-district, respectively. From Meurebo sub-district profenofos 0.74 mg kg-1 was found. All detected pesticide residues samples are still below the maximum residue limit (MRL) set by the Government of Indonesia and CODEX Alimentarius.

Outcomes with venetoclax in myelodysplastic syndromes: A systematic review.

e19035 Background: Myelodysplastic syndromes (MDS) are clonal hemopoietic disorders causing cytopenia(s) and increased risk of transformation to acute myeloid leukemia (AML). Venetoclax is a B-cell lymphoma-2 (BCL-2) inhibitor that induces apoptosis in malignant cells. The FDA has approved venetoclax for the treatment of newly diagnosed adult AML patients unfit for intensive chemotherapy and its utility in MDS is being explored. We present a systematic review aimed to evaluate outcomes with venetoclax in MDS patients. Methods: We performed a literature search on 3 databases (Pubmed, Cochrane, and Clinicaltrials.gov) following the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) guidelines. We screened 62 studies using the Mesh terms ‘myelodysplastic syndrome’ and ‘venetoclax’. After excluding review, duplicate and non-relevant articles, 8 studies were included in the analysis. Quality evaluation was done using the NIH quality assessment tool. Pooled analysis was done using the ‘meta’ package (Schwarzer et al, R programming language) and proportions with 95% confidence intervals (CI) were computed. The Inter-study heterogeneity among the studies was assessed using the Q statistic proposed by Cochrane and the I2 index introduced by Higgins and Thompson. Results: We identified 313 patients from 8 studies (1 prospective, 3 retrospective, 2 ex-vivo, 2 clinical trials). Venetoclax was used in combination with hypomethylating agents (HMA) in 6 studies and with other agents in 2 studies. Most patients were pre-treated. Complete response (CR) with or without hematological recovery (CRi) was reported in 45% of patients (95% CI 0.26-0.66, I2= 90%, n = 292). Stable disease (SD) was seen in 18% patients (95% CI 0.08-0.37, I2= 85%, n = 223). Overall response rate (ORR) was 51% (95% CI 0.31-0.70, I2= 89%, n = 122). Hematopoietic stem cell transplant (HCT) was performed in 22% patients (95% CI 0.09-0.44, I2= 75%, n = 122). Twenty percent patients (95% CI 0.15-0.28, I2= 0%, n = 149) died. Common adverse events seen with venetoclax were cytopenias, gastrointestinal side effects and infections, often requiring drug interruption or dose. Conclusions: Despite heterogeneity of available literature, venetoclax seems to be a promising agent in treatment of MDS patients. Prospective clinical trial data is needed to ascertain safety and efficacy of venetoclax in MDS and impact on overall survival, in particular, in patients proceeding to HCT.[Table: see text]

Anticancer Potential of L-Asparaginase: An Overview

Asparaginase, derived from microbial origin hydrolyses L-asparagine to L-aspartic acid. The enzyme finds principal use in the treatment of Acute Lymphoblastic Leukemia during childhood that primarily occurs between two to ten years of age. L-Asparaginase finds its use in management of haemopoietic disorders especially in pediatrics that is caused due to proliferation and enlargement of lymphoblast in bone marrow and in blood as well as other part of the body. L- Asparaginase from bacterial sources exhibit quaternary and tertiary structural forms. However for using it in therapeutic and clinical application it should not generate any fatal allergic reaction to the patient. Such effects can occur due to the enzyme associated L-Glutaminase activity and also due to the endotoxins from bacteria in enzyme preparations. Therefore, with the recent development in biotechnology with respect to production and purification techniques it is possible to get pure L- asparaginase from microbial origin. The present article provides an insight into the mechanism of action of L-Asparaginase as an anticancer agent and its industrial applications.

Serum Lipidome and Glucose Metabolism Uncover Vulnerabilities in Myelodysplasia and Secondary Acute Myelogenous Leukemia

Background Secondary acute myelogenous leukemia (sAML) is highly aggressive and molecularly heterogenous. Previous reports have demonstrated differential serum lipid expression among hemopoietic malignancies (i.e AML, myelodysplastic syndrome (MDS) and acute lymphoblastic leukemia (ALL). Such a distinct serum lipidome among hemopoietic disorders suggest "unique oncogenic metabolic addiction" of leukemia initiating cells (LICs). MDS to AML transition is characterized by clonal evolution [i.e severe aneuploidy, high P53 frequency etc.] and chemorefractoriness. However, mechanism(s) for transformation are not entirely understood. In this study, we investigate serum lipidome and "proxy" of glucose [HbA1C] metabolism among: (1) low risk, (2) high risk MDS and (3) sAML patients (pts) to identify features that may suggest disease specific vulnerabilities. Methods After IRB approval, 214 pts were selected for analysis. ANOVA was used to detect differential expression of total cholesterol (TC), low density lipoprotein (LDL), high-density lipoprotein (HDL), and triglycerides (TAG) among low risk, high risk and sAML pts. Multivariate linear regression model was performed to evaluate the independent effect of confounders on relevant lipid modifications observed among subtypes of MDS and sAML pts. SAS software was used for data processing. Results 69/214 (32.2%), 26/214 (12.1%), 35/214 (16.3%), 35/214 (16.3%), 28/214 (13.08%), 21/214 (9.8%) were sAML, very low (VLR), low (LR), intermediate (I), high (H) and very high risk (VHR) MDS. Median age (range) was 68 y (42-91), 68 y (22-87), 70 y (55-91), 76 (53-88), 72 y (60-85) and 74 y (54-84), for sAML, VLR, LR, I, H and VHR, p=0.0001. In sAML, VLR, LR, I, HR and VHR MDS TC was 177 mg/dL, 150 mg/dL, 126 mg/dL, 134 mg/dL, 125 mg/dL and 122 mg/dL, p=0.0001; LDL was 113.4 mg/dL, 81.3 mg/dL, 67.8 mg/dL, 72 mg/dL, 63.2 mg/dL and 70.8 mg/dL, p=0.0001; HDL 35.8 mg/dL, 39.2 mg/dL, 36.1 mg/dL, 40 mg/dL, 33.7 mg/dL, 32.05 mg/dL, p=0.40; TAG 134 mg/dL, 140 mg/dL, 109 mg/dL, 120 , mg/dL 134 mg/dL, 98 mg/dL, p=0.1. Given "similarities" among MDS subgroups, "MDS lipid data" was compared vs sAML. Median TC, LDL, HDL and TAG among sAML vs MDS pts was 179.1 mg/dL vs 130.4 mg/dL, p=0.0001; 113 mg/dL vs 70.1 mg/dL, p=0.0001; 38.6 mg/dL vs 36.7 mg/dL, p=0.5; 142 mg/dL vs 116.6 mg/dL, p=0.056, respectively. 20/89 (22.7%) vs 69/89 (77.53%) sAML vs MDS pts, respectively were diabetic, p=0.002, OR =2.62; 95% CI 1.40-4.90. However, data extracted from 82 pts (16 sAML and 66 MDS, each) demonstrated that hemoglobin A1C (HbA1C) was 7.84% vs 6.34%, in sAML vs MDS, respectively, p=0.009. Multivariate linear regression accounting for age, blast count, BMI, DM status and HbA1C showed that higher TC, LDL and HbA1C were independent predictors of sAML. Conclusions While this study is restricted to demonstrate distinct lipid and metabolic profile between sAML and MDS, our results suggest blastic phase dependency on higher serum cholesterol and possibly higher advanced glycosylation. As compared to MDS, our data highlights the possibility that sAML induces serum Lipidome modifications that may "hijack endogenous lipogenesis" to favor TC, LDL and possibly TAG production. Additionally, despite larger number of patients is needed to confirm our results, higher HbA1C potentially initiates advanced glycosylation facilitating blastic conversion in MDS pts. Disclosures Rivero: Incyte: Membership on an entity's Board of Directors or advisory committees; agios: Membership on an entity's Board of Directors or advisory committees; celgene: Membership on an entity's Board of Directors or advisory committees.

Excess in Atherosclerotic and Inflammametabolic Diseases Are Differentially Expressed in Myelodysplasia and Are Highly Dependent on Age, R-IPSS and Ethnicity

Background Myelodysplastic syndrome (MDS) is an inflammatory heterogeneous group of myeloid disorders with variable risk for acute myeloid leukemia (AML) conversion. Recent reports highlight that mutations [i.e. TET2, DNMT3A, AXSL1] configurate not only risk for malignant conversion in “healthy elderly individuals” but also ‘directly’ participate in coronary artery disease [CAD] development. TET2 clonal hematopoiesis [TET2 CHIP] accelerates atherosclerosis in mice via monocytic inflammatory amplification. Indeed, germ-line DNMT3A human disease phenocopy obesity and neurocognitive abnormalities suggesting that a more complex, but still poorly characterized cardiometabolic risk, probably originates from patient predisposition and somatic mutation acquisition. We seek to originate a cross-sectional “proof of concept” for quantifying the association between myelodysplasia [a clonal disorder initiated by CHIP mutations] and several systemic inflammatory conditions [i.e. CAD, peripheral vascular disease (PVD), obesity, depression and COPD].Methods: A cross-sectional analysis of a nationally-representative sample of inpatient hospitalizations in the United States between 2007 and 2015 [750,249 MDS-related hospitalizations, or 83,361 hospitalizations per year] was performed to identify co-occurring inflammatory diseases during hospitalizations in patients older than 35 years, in which MDS was documented. Given the inflammatory nature of the disease, subgroups were defined to investigate the prevalence of individual inflammatory domains (per 1,000 MDS-related hospitalizations) according to Revised-International Prognostic Score System [R-IPSS]. Subgroup analysis was performed for patients based on age (above and below 65 years), and race/ethnicity. Prevalence and confidence intervals [CI] were obtained for each inflammatory condition. “Relevant” inflammatory domains were considered statistically significant based on non-overlapping 95% CIs.Results: In examined admissions, prevalence for CAD in (Low-risk) LR- MDS age <> 65 was 12.2 vs 24.7, PVD 21.8 vs 60.2, [Fig 1 A and B] obesity 88.1 vs 51.7, major depression 14.6 vs 5.3, COPD 42.1 vs 70.7, p=0.0001 for all comparisons [Fig 1 B and C]. Conversely, prevalence for CAD in (High-risk) HR MDS age <> 65 was 9.1 vs 17.3, p=0.04; PVD 14.5 vs 43.2, p=0001; obesity 73.7 vs 42.2, p=0.003 major depression 5.1 vs 6.9, p=0.04 and COPD 23.0 vs 34.6, p=0.05. Differential prevalence expression for CAD was observed between non-Hispanic Whites (W) and Hispanics (H) [22 vs 28, p=0.02]. PVD was more prevalent, in both W vs H [64.6 and 52.2, p= 0.0002] and NH-Blacks/African Americans (AA) vs H [63.1 vs 52.2, p= 0.009]. Obesity was more prevalent among AA and H [64.1 and 62.9, respectively] than in W [51.9, p=0.009 for W vs AA and p=0.0002 for W vs H]. Similar expression for diabetes was observed between H vs W [404 vs 282, p<0.0001], H vs AA [404 vs 364, p=0.0001], and AA vs W [364 vs 282, p=<0.0001]. COPD was dominant among W and AA [73.8 and 78.4] as compared with H [50.4, p=0.0001 for H vs W and H vs AA]. Idiopathic pulmonary fibrosis was dominant in W and H [1.8 and 1.9] in comparison to AA [0.9, p=0.04 AA vs H and p=0.02 AA vs W]. Major depression prevalence was observed in AA, W and H in 5.4 vs 9.5 vs 14.3, respectively, p=0.0001 H vs AA and H vs W].Conclusions. Prevalence of systemic inflammatory diseases in MDS are associated with age, disease biology [Low vs High risk] and ethnicity. Cardiopulmonary complications, such as PVD+ COPD are dominant in W and AA. Lower prevalence for PVD and COPD, but high rates for CAD, depression, diabetes and IPF in H vs W and AA suggest that ethnicity could differentially drive cardiometabolic and inflammametabolic phenotypes when co-occurring with clonal hemopoietic disorders such as MDS. DisclosuresNo relevant conflicts of interest to declare.

Clinical Understanding of Inflammation, Inducing Thrombogenesis, Respiratory Impairment, and Cardiorenal Syndrome in Patients With Pulmonary and Coincident Cardiac Dyspnea

SESSION TITLE: Pulmonary Vascular Disease I SESSION TYPE: Original Investigation Poster PRESENTED ON: Wednesday, October 26, 2016 at 01:30 PM - 02:30 PM PURPOSE: The study aims to investigate understanding of clinical pathophysiology of primary inflammation in pulmonary and coincident cardiovascular dyspnea patients(Pts). METHODS: 111 Pts, 67 males and 40 females,mean age 66 years and 40, matched control,normal individuals underwent: 1.Biomarkers serum evaluation (CRP and pro-inflammatory cytokines,D-dimers, NT-ProBNP, Tr-I, EPO, MMP-2,PCT, 2. Physical examination, ECG, Chest X-ray and CT 3. Echocardiography 4. PFTs and ABGs 5. Blood tests. RESULTS: Resulted abnormal values of serum Biomarkers,echocardiographic and Doppler indices,PFTs,ABGs and Blood tests presented significant correlations as follows: CRP with A) D-dimers B) PCO2 C) NT-ProBNP D) EF E) LVID. Furthermore, A) D-dimers with A1) RVSP,correlated to LAD and LVID, A2) RVSP correlated with P(A-a)O2 and -FVC and -TLC, A3) PCO2. B) PCO2 with NT-ProBNP, FEV1 and Na, correlated with left ventricular E-Wave . C) NT-ProBNP with EF, PCO2 and C1) Tr-I, C2) ASH, C3) Hb, C4) Urea and Creatinine, C5) Pleural NT-ProBNP. However, C1) Tr-I correlated with left ventricular E/A ratio, MMP-2, EPO, - FVC and -TLC, - Na and Creatinine. C2) ASH with EF, Urea and Creatinine. C3) Hb with Pleural NT-ProBNP, TLC and PCT,correlated with MMP-2, Tr-I, EPO and PCO2. D) EF with FVC and Tr-I. E) LVID with LAD, RVSP, RVID, Creatinine and Urea. CONCLUSIONS: Reported abnormal results and significant correlations arise originally from an inflammatory process,expressed by CRP levels, introducing various pathways of clinical pathophysiology, correlated with D-dimers, PCO2 , NT-ProBNP, EF and LVID. 2. All indices lead either to thrombogenesis and/or pulmonary, myocardial and renal dysfunction, followed by disarrangement of haemopoiesis and infective complications. 3. Inflammatory thrombogenesis also seems developing on the grounds of cardiovascular apoptosis and remodelling. 4. The reported molecular and interstitial disarray phenomena, combined with pulmonary and renal impairment ,further worsen haemopoietic disorders and late infection, all possibly creating a dynamic early-late inflammation-infection pathophysiology, carrying on thrombogenesis, pulmonary and cardio-renal dyspnea. CLINICAL IMPLICATIONS: Further studies should be carried out to confirm the reported abnormal data with their significant correlations as well as clarify understanding of early inflammation and late infection, both probably promoting thrombogenesis, respiratory impairment and cardio-renal syndrome. DISCLOSURE: The following authors have nothing to disclose: Ioannis Angomachalelis, Stavros Tryfon, Nestor Angomachalelis No Product/Research Disclosure Information

Haemato-biochemical disorders associated with nodavirus like-agent in adult leaping mullet Liza saliens (Risso, 1810) in the Caspian Sea.

Betanoda virus is an emerging problem in several marine fish species in various geographic areas all over the world. In recent years, mullets stock of the Caspian Sea decreased dramatically and a betanoda virus like-agent was introduced as the cause of mullet's mortality. The main objective of the present study is to compare hemato-biochemical parameters in healthy and infected sharpnose mullets (Liza saliens) to betanoda virus like-agent. The adult sharpnose mullets (34 clinically affected fish+34 apparently healthy fish) were captured from the southeast the Caspian Sea from 2012 to 2013. All of the captured fish were 4 or 5years old. The main clinical sings of infected fish were lethargy, severe abdominal distention, abnormal swimming and hyperinflation of swim bladder. The results showed that the weight and total length of infected fish were significantly lower than the healthy fish. The results of histopatological evaluation and indirect florescent antibody test were confirmed the presence of a betanoda virus-like agent in infected fish. The diseased fish were severely anemic (hypochromic macrocytic anemia) and had a serious haemopoietic disorders. The anemia associated with a sever leukopenia, a significant rise of neutrophils and immature neutrophils and a significant decries of lymphocytes percentage. Total protein, albumin and total immunoglobulin levels were significantly reduced in the serum of infected fish, while the activities of aspartate aminotransferase and alanine aminotransferase significantly increased when compared to the healthy fish. These results suggested that the feeding of the infected fish to betanoda virus like-agent were disturbed and it could be cause haemato-biochemical disorder and mortality of the fish in Iranian water of the Caspian Sea.

Combined overexpression of WT1 and BAALC genes May Predict AML evolution in MDS Patients

Background and aimsMyelodisplastic syndromes (MDS) are a group of hemopoietic disorder characterized by an impaired blood cell production, morphologic dysplasia and peripheral cytopenias; they are the most common hematologic neoplastic disorder and its diagnosis relays on morphologic evaluation, associated to a karyotypic assay.In order to predict the outcome of patients affected from these disorders, knowing that the order of survival can be extremely variable, several prognostic index were developed such as International Prognostic Score Sistem (IPSS) or the most usefull WHO-Prognostic Score Sistem (WPSS).On the contrary of acute leukemia, these disorders have not a biomolecular profile evalutation of intrinsic markers able to stratify patients in different prognostic risk groups.The aim of our study is to assess the risk of leukemic evolution, in MDS patients, on the basis of the levels expression of WT1 and BAALC at disease diagnosis, and to evaluate the leukemia free survival (LFS) at 6-12-24-36 months of follow up, among the different risk category according to IPSS and irrespectively of treatment. Materials and methodIn five years we analized 102 patients with a diagnosis of MDS divided according to the WHO classification such as follows: 38 AR, 1 AR with del(q5), 21 aREB-1, 23 AREB-2, 3 chronic myelomonocitic leukemia, 1 RARS, 1 MDS, 11 RCMD, 1 5q- syndrome, 2 suspected MDS.According to IPSS 58 belonged to the low risk category, 21 to the intermediate-1, 23 to the intermediate-2/high risk.Cytogenetic assay showed 20 people with an abnormal karyotype, 8 of them fallen into the high risk class and 12 into the intemediate risk.Low risk and intermediate-1 patients were treated only with supportive care; high risk patients were treated with hypomethylating agents. Iron chelation were used when necessary. Lenalidomide was used in the only case of 5 q- syndromeSamples of bone marrow were analized with Real-Time quantitave PCR and levels of WT1 and BAALC expression were determinated.Molecular datas were analized with X-square Test and a significant association was recorded between overexpression of the genes evaluated (WT1 higher than 100 copy numbers and BAALC higher than 1000 copy numbers) and the probability of develop acute myeloid leukemia ( AML ). ResultsNine out of 102 patients showed an isolated WT1 hyperexpression (3 of them developed an AML ), in 15 cases we reported an isolated BAALC overexpression (3 of them developed an AML ), while 13 out 18 patients ( 72% ) with combined WT1 and BAALC overexpression developed AML within an average time of 6 months; instead only 5% of patients, which expressed low levels of WT1 and BAALC, developed AML within the interval of observation.In particulary a combined high expression of WT1 and BAALC were strongly associated with an high risk to develop leukemia and a short LFS, especially in INT-1 subset.After that we calculated the LFS, divided for the risk category at 6-12-24-36 months of follow up.Patients with combined overexpression of WT1 and BAALC showed a LFS of 40% at 6 months of follow up and 0% at 24 months. ConclusionMDS have a great variable survival, and the current approach to these diseases relays on morphological evaluation, karyotypic assay and need of transfusional support; gene expression could be a promising system to predict the prognosis in these patients.Analysis of gene expression, which belong to AML evaluation, allows to divide patients in several risk groups; furthermore is not the single gene evaluation that is more predictable but a combined assay.With this method, which seems to be more realiable than IPSS, we could find that a great percentage of patients with levels of WT1>100 and BAALC >1000, indipendently from karyotypic status and treatment, developed AML and have a shorter LFS than the population with WT1 <100 and BAALC < 1000. [Display omitted] Disclosures:No relevant conflicts of interest to declare.

Spontaneous morphological and cytogenetic remission of MDS: A case report and review of the literature

Introduction: Myelodysplastic syndromes (MDS) are a group of haemopoietic stem cell disorders characterised by cytopenias and dysplasia in one or more myeloid cell lines with a risk of developing acute leukaemia. MDS can occur as a primary event or following exposure to chemotherapy or radiotherapy. Spontaneous remission can occur but is rare. Case Report : In this case report, a 39­year­old female developed MDS with a cytogenetic abnormality t (1;7) during treatment with cyclophosphamide for Wegener’s granulomatosis. A spontaneous morphological and cytogenetic remission occurred six years after initial diagnosis. Conclusion: Case reports of spontaneous remission in patients with primary MDS occasionally appear in the literature however there are very few reports concerning remission of therapy related MDS. This case is unusual as the patient achieved complete haematological and cytogenetic remission of treatment related MDS on a reduction of cyclophosphamide.

MicroRNA-146a and hemopoietic disorders

MicroRNAs (MiRNAs) are a class of small non-coding regulatory RNAs that repress protein expression at the posttranscriptional level and play important roles in hematopoiesis processes. MiR-146a is a miRNA that is thought to regulate physiological and pathophysiological pathways in hematopoietic cells. In this review, we focus on recent progress in analyzing the functional roles of miR-146a in normal hematopoiesis and hematopoietic disease. We suggest that manipulation of miR-146a expression may represent a potential new therapy for several hematopoietic diseases, and may further serve as a biomarker for diagnosis, prevention, and treatment of such disease.

Mutations in the transmembrane and juxtamembrane domains enhance IL27R transforming activity

Cytokines and their receptors regulate haemopoiesis by controlling cellular growth, survival and differentiation. Thus it is not surprising that mutations of cytokine receptors contribute to the formation of haemopoietic disorders, including cancer. We recently identified transforming properties of IL27R, the ligand-binding component of the receptor for interleukin-27. Although wild-type IL27R exhibits transforming properties in haemopoietic cells, in the present study we set out to determine if the transforming activity of IL27R could be enhanced by mutation. We identified three mutations of IL27R that enhance its transforming activity. One of these mutations is a phenylalanine to cysteine mutation at residue 523 (F523C) in the transmembrane domain of the receptor. The two other mutations identified involve deletions of amino acids in the cytoplasmic juxtamembrane region of the receptor. Expression of each of these mutant IL27R proteins led to rapid cytokine-independent transformation in haemopoietic cells. Moreover, the rate of transformation induced by these mutants was significantly greater than that induced by wild-type IL27R. Expression of these IL27R mutants also induced enhanced activation of JAK (Janus kinase)/STAT (signal transducer and activator of transcription) signalling compared with wild-type. An activating deletion mutation of IL27R enhanced homodimerization of the receptor by a mechanism that may involve disulfide bonding. These transforming IL27R mutants displayed equal or greater transforming activity than bona fide haemopoietic oncogenes such as BCR-ABL (breakpoint cluster region-Abelson murine leukaemia viral oncogene homologue) and JAK2-V617F. Since IL27R is expressed on haemopoietic stem cells, lymphoid cells and myeloid cells, including acute myeloid leukaemia blast cells, mutation of this receptor has the potential to contribute to a variety of haemopoietic neoplasms.

The Use of Autologous Peripheral Blood Stem Cells as a Hemopoietic Support during Polychemotherapy of Children with Soft Tissue Sarcomas

Intensification of cycle polychemotherapy in disseminated tumors considerably improves the efficiency of complex treatment. Reinfusion of peripheral blood stem cells as a factor of replacement treatment during hemopoietic suppression or disorders is now becoming more and more promising.

The role of soluble and cell-surface expressed 4-1BB ligand in patients with malignant hemopoietic disorders

The TNFR family member 4-1BB and its ligand 4-1BBL are involved in the costimulation of T-cells and tumor-derived soluble (s)4-1BBL may influence the interaction of malignant cells with the immune system. Here, we report that cell-surface-expressed (c)4-1BBL can be expressed on mononuclear blood cells from patients with acute myeloid leukemia (AML) (n = 35), myelodysplasia (n = 5) or non-Hodgkin lymphoma (n = 11) and can be coexpressed on varying proportions of lymphoid or myeloid malignant cells and on dendritic cells differentiated from AML-blasts. Direct correlations between c- and s4-1BBL were not found in the investigated cases. Up to now expression of 4-1BBL has not been described on primary myeloid malignant cells, but only on malignant cells of lymphoid or solid tumor origin or on tumor cell lines. With our work we further contribute to the understanding of the potential role of c/s4-1BBL in immune reactions and its influence on the interaction of tumor and immunoreactive cells.

Busulfan and Cyclophosphamide for Hemopoietic Stem Cell Transplantation from Related and Unrelated Donors in Patients with Myelodysplastic Syndrome

The myelodysplastic syndromes (MDS) are clonal hemopoietic disorders, characterized by ineffective hemopoiesis resulting in single or multilineage peripheral blood cytopenias, dysplastic morphology in single or multiple lineages, and, in many patients, clonal cytogenetic abnormalities. Hematopoietic stem cell transplantation is currently the only therapeutic modality that is potentially curative. A total of 17 patients (aged 16–40 years; median, 26 years) with MDS were treated with busulfan (BU 3.2 mg/kg/d, −7 d ‘−4 d,) plus cyclophosphamide (CY, 60 mg/kg, −3 d ‘−2 d)and hemopoietic stem cell (HSC) transplantation from related (n =5) or unrelated donors (n =12). At the time of transplantation, 13 patients had less than 5% myeloblasts in the marrow, and 4 patients had more advanced disease. Mycophenolate mofetil combined with cyclosporin A and methotrexate was used for prevention of acute graft-versus-host disease after transplantation. Lipo prostaglandin E1 was used in prophylactic regimen for hepatic venoocclusive disease. All but 2 evaluable patients had engraftment. Neutrophil count began to be higher than 0.5×109/L on 15 days after BMT. Platelet count began to be higher than 20×109/L on 18 days after BMT. The Kaplan-Meier estimates of 3-year relapse-free survival (RFS) were 57%, and only one patient relapsed after six months of transplantation. Non relapse mortality (NRM) at 100 days was 12%. Thus, a targeted BUCY regimen provided effective transplant conditioning for patients with MDS receiving transplants from HLA identical siblings or alternative donors.