scholarly journals Hemoglobin Variants Influence Plasmodium Falciparum Sexual Differentiation

Blood ◽  
2021 ◽  
Vol 138 (Supplement 1) ◽  
pp. 965-965
Author(s):  
Bethany Flage ◽  
Matthew Dent ◽  
Jesús Tejero ◽  
Solomon Fiifi Ofori-Acquah
Keyword(s):  
Plasmodium Falciparum ◽  
Sexual Differentiation ◽  
Conflicts Of Interest ◽  
Causative Factor ◽  
Hemoglobin A ◽  
Conversion Rates ◽  
Significant Difference ◽  
Hb S ◽  
Hemoglobin Variants ◽  
Digestion Efficiency

Abstract It has long been recognized that individuals who express variations of the hemoglobin-A (HbA) protein experience less severe malaria disease. As malaria remains to be one of the most significant infectious diseases in history, this human adaptation has led to the persistence of HbA variants (HbVARs) in the population. The intricate lifecycle of the parasite which causes the most cases of clinical malaria, Plasmodium falciparum, relies on both asexual and sexual reproductive cycles, with host to vector transmission reliant on sexual stage gametocyte formation. Multiple epidemiological studies have shown that HbVARs may influence gametocyte production during P. falciparum infection, with greater gametocyte numbers reported in individuals with hemoglobin variant containing erythrocytes (Hb VAR-Ery) when compared to hemoglobin A containing erythrocytes (Hb A-Ery). Here we provide experimental support for these studies by showing significantly higher sexual differentiation rates among parasites grown in Hb S containing erythrocytes (Hb S-Ery) obtained from sickle cell patients than those differentiated in Hb A-Ery (p=0.038). Because the digestion of hemoglobin is such an integral part of the intraerythrocytic cycle, we then sought to determine whether there was a difference between the hydrolysis efficiencies of HbA and other hemoglobin variants (HbVAR). By using a prominent recombinant P. falciparum hemoglobinase we found the hydrolysis efficiency of HbA to be significantly (p=0.0058) more efficient after 24 hours compared to a HbVAR sample containing mixed amounts of HbA, HbF, and HbS. To further determine whether there is a link between hemoglobin digestion efficiency and sexual differentiation, we therapeutically inhibited the hemoglobin digestion and hemozoin formation process in a culture of P. falciparum using sub-optimal doses of chloroquine diphosphate. We found a significant difference (p<0.001) among gametocyte conversion rates between treated and non-treated cultures, as well as a moderate negative correlation between hemozoin formation and gametocyte conversion rate (Pearson r=0.72, p=0.008). Gene expression analysis also revealed patterns of expression that were consistent with increased gametocytogenesis. We conclude that hemoglobin type plays a significant role in the process of sexual conversion in P. falciparum. Though further studies should be completed in order to confirm these results, these findings may suggest hemoglobin digestion efficiency as a causative factor for sexual differentiation. As individuals with hemoglobinopathies make up approximately 7% of the global population, and malaria infection rates have been shown to differ depending on these genetic dynamics, these findings may support the creation of targeted initiatives to reduce transmission specifically in areas where there is a high percentage of hemoglobinopathy carriage. Disclosures No relevant conflicts of interest to declare.

Physiologic differences in hemoglobin variants

1965 ◽  
Vol 208 (1) ◽  
pp. 198-202 ◽  
Author(s):  
Robert B. Thompson ◽  
Richard L. Warrington ◽  
Warren N. Bell
Keyword(s):  
Cord Blood ◽  
Whole Blood ◽  
Fold Increase ◽  
Twofold Increase ◽  
Hemoglobin A ◽  
Hb E ◽  
Hb S ◽  
Hemoglobin Variants

Studies were carried out on whole blood, concentrated hemoglobin solutions, and isolated hemoglobin fractions as related to oxygen association. In hemoglobins Ao, A1, Dalpha, GPhiladelphia, and C, no difference was found in the oxygen association curve. Hemoglobins A1, B2, and Lepore showed a 2½ fold increased oxygen association over normal Hb-A. Hemoglobin solutions from cord blood showed no difference from Hb-A, whereas cord blood showed a twofold increase. Hb-H and Hb-Barts showed identical oxygen association curves with a 10- to 12-fold increase over Hb-A; no heme-heme interaction and no Bohr was found with Hb-H or Barts. Hb-E and Hb-S showed a slight decrease in oxygen equilibria over hemoglobin A.

scholarly journals Donor Lymphocyte Infusion Prevented Late Post-Transplant Rejection in 30 Patients with Thalassemia Major

Blood ◽  
2018 ◽  
Vol 132 (Supplement 1) ◽  
pp. 2344-2344
Author(s):  
Jianyun Liao ◽  
Xiaoting Liu ◽  
Yuelin He ◽  
Zhiyong Peng ◽  
Huaying Liu ◽  
...  
Keyword(s):  
Conflicts Of Interest ◽  
Immunosuppressive Agents ◽  
Thalassemia Major ◽  
Mixed Chimerism ◽  
Hematopoietic Stem ◽  
Post Transplant ◽  
Conversion Rates ◽  
Significant Difference ◽  
Group 2 ◽  
Group 1

Abstract OBJECTIVE:To investigate the role of donor lymphocyte infusions (DLI) in preventing post-transplant graft rejection (GR) in hematopoietic stem cell transplantation (HSCT) for patients with β-thalassemia major (TM); METHODS: A total of 50 patients with TM who had mixed chimerism (MC) after HSCT from June 2011 to June 2016 was divided into 4 groups. The group 1 included the patients with grade I MC ratio of 90%-95% (n=20) but not DLI; Group 2 patients had same grade I MC ratio as Group 1 and received DLI (n=11); Group 3 had grade II MC ratio of 75%-90% (n=13); Group 4 had grade III MC of <75% (n=6). Group 1 received gradually reduced immunosuppressive agents; Groups 2 to 4 received DLIs of gradually increasing dose after reduction of immunosuppressive agents; RESULTS: (1) Full donor chimerism (FDC) conversion rates in the groups 1 to 4 were 100%, 100%, 85%, and 66.7%, respectively; The incidence of graft versus host disease (GVHD) in groups 1 to 4 was 15%, 27.3%, 46.2%, and 33.3%, respectively; (2) There was no significant differeces as comparing incidence of GVHD in group 1 and in group 2 (15% vs. 27.3%, P = 0.638); (3) A significant difference was found in comparing patients receiving initial DLI at ≤180 and > 180 days and the incidence of GVHD was 50% vs 10% (P=0.049); (4) Comparing the accumulated doses of DLI >1×107/kg with ≤1×107/kg, the incidence of GVHD was 50% vs. 27.3% (P=0.142); CONCLUSION: 1. The optimal choice for post-transplant MC grade 1 patient is only observation and decreasing the immunosuppressive drugs; 2. For grade 2-3 patients with high risk of rejection, the fractional increasing dose DLI can effectively reverse the unstable chimera to FDC; 3. Post-DLI GVHD was associated with DLI timing, and the incidence of GVHD was higher in patients who received DLI at ≤180 days after transplantation. Disclosures No relevant conflicts of interest to declare.

scholarly journals Comparison of leucocyte profiles between healthy children and those with asymptomatic and symptomatic Plasmodium falciparum infections

2020 ◽  
Vol 19 (1) ◽  
Author(s):  
Diana Ahu Prah ◽  
Linda Eva Amoah ◽  
Matthew P. Gibbins ◽  
Yaw Bediako ◽  
Aubrey J. Cunnington ◽  
...  
Keyword(s):  
T Cells ◽  
Plasmodium Falciparum ◽  
Peripheral Blood ◽  
Parasite Load ◽  
Study Groups ◽  
Healthy Children ◽  
Peripheral Cell ◽  
Peripheral Blood Leucocyte ◽  
Cell Counts ◽  
Significant Difference

Abstract Background The immune mechanisms that determine whether a Plasmodium falciparum infection would be symptomatic or asymptomatic are not fully understood. Several studies have been carried out to characterize the associations between disease outcomes and leucocyte numbers. However, the majority of these studies have been conducted in adults with acute uncomplicated malaria, despite children being the most vulnerable group. Methods Peripheral blood leucocyte subpopulations were characterized in children with acute uncomplicated (symptomatic; n = 25) or asymptomatic (n = 67) P. falciparum malaria, as well as malaria-free (uninfected) children (n = 16) from Obom, a sub-district of Accra, Ghana. Leucocyte subpopulations were enumerated by flow cytometry and correlated with two measures of parasite load: (a) plasma levels of P. falciparum histidine-rich protein 2 (PfHRP2) as a proxy for parasite biomass and (b) peripheral blood parasite densities determined by microscopy. Results In children with symptomatic P. falciparum infections, the proportions and absolute cell counts of total (CD3 +) T cells, CD4 + T cells, CD8 + T cells, CD19 + B cells and CD11c + dendritic cells (DCs) were significantly lower as compared to asymptomatic P. falciparum-infected and uninfected children. Notably, CD15 + neutrophil proportions and cell counts were significantly increased in symptomatic children. There was no significant difference in the proportions and absolute counts of CD14 + monocytes amongst the three study groups. As expected, measures of parasite load were significantly higher in symptomatic cases. Remarkably, PfHRP2 levels and parasite densities negatively correlated with both the proportions and absolute numbers of peripheral leucocyte subsets: CD3 + T, CD4 + T, CD8 + T, CD19 + B, CD56 + NK, γδ + T and CD11c + cells. In contrast, both PfHRP2 levels and parasite densities positively correlated with the proportions and absolute numbers of CD15 + cells. Conclusions Symptomatic P. falciparum infection is correlated with an increase in the levels of peripheral blood neutrophils, indicating a role for this cell type in disease pathogenesis. Parasite load is a key determinant of peripheral cell numbers during malaria infections.

Commitment to sexual differentiation in the human malaria parasite, Plasmodium falciparum

Parasitology ◽  
2000 ◽  
Vol 121 (2) ◽  
pp. 127-133 ◽  
Author(s):  
T. G. SMITH ◽  
P. LOURENÇO ◽  
R. CARTER ◽  
D. WALLIKER ◽  
L. C. RANFORD-CARTWRIGHT
Keyword(s):  
Plasmodium Falciparum ◽  
Sexual Differentiation ◽  
Malaria Parasite ◽  
Human Malaria Parasite ◽  
Human Malaria ◽  
Parasite Plasmodium ◽  
Parasite Plasmodium Falciparum

scholarly journals 1241. PfSPZ Vaccine Administered by Direct Venous Inoculation to Prevent Plasmodium falciparum Malaria is as Safe as Normal Saline – a Meta-analysis of 12 Randomized Controlled Clinical Trials

2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S639-S640
Author(s):  
L W Preston Church
Keyword(s):  
Plasmodium Falciparum ◽  
Random Effects ◽  
Normal Saline ◽  
Meta Analysis ◽  
Laboratory Data ◽  
Plasmodium Falciparum Malaria ◽  
Sub Saharan Africa ◽  
Sensitivity Analyses ◽  
Double Blind ◽  
Significant Difference

Abstract Background Sanaria’s PfSPZ Vaccine prevents Plasmodium falciparum (Pf) infection transmitted in the field and by controlled human malaria infection. Safety of PfSPZ Vaccine has been demonstrated in 12 randomized, double-blind, placebo-controlled trials (RCT) varying in regimen from 3 to 5 doses over 4 to 20 weeks and in size from 18 to 332 subjects in adults in the US and EU and 5-month to 65-year-olds in 5 countries in sub-Saharan Africa. This study was conducted to analyze solicited adverse event (AE) and laboratory data by random effects meta-analysis. Methods PfSPZ Vaccine is composed of radiation-attenuated, aseptic, purified, cryopreserved Pf sporozoites (SPZ) administered by direct venous inoculation (DVI). Normal saline (NS) is always the placebo. Data from all completed RCTs were included as either age &gt; 18 years (n=598) or age 5 months to 17 years (n=641). Any subject receiving at least one dose was included. A random-effects model was used to study vaccine safety and I2 to evaluate heterogeneity. Analysis was performed for any systemic solicited AE and for the most frequently observed AEs and laboratory abnormalities. Sensitivity analyses were performed by removal of trials with zero events to evaluate potential bias. Results When examined individually, only 1 trial had a significant difference between PfSPZ Vaccine and NS for any AE (myalgias in adults). In the adult meta-analysis, there was no difference in the random effects risk ratios (RR) for having any vaccine-related AEs (1.40, 95% confidence interval (CI) 0.88-2.28), or for fever (0.75, 0.24-2.35), headache (1.23, 0.74-2.02), fatigue (0.72, 0.19-2.54), or myalgia (1.09, 0.26-4.68). In the pediatric meta-analysis there was no difference between the RR for PfSPZ Vaccine and NS for any AE (0.84, 0.59-1.18) or for fever (1.09, 0.44-2.69). No significant differences in the most common grade 2 or higher laboratory abnormalities – declines in hemoglobin, neutrophil or platelet count – were detected. Sensitivity analysis did not change the results. Conclusion There was no difference in risk for AEs or lab abnormalities between PfSPZ Vaccine and NS, indicating that PfSPZ Vaccine administered by DVI was extremely safe and well tolerated in 5-month- to 65-year-olds. Disclosures LW Preston Church, MD, FIDSA, Sanaria Inc. (Employee)

Urinary Growth Hormone Measurements in Children with Renal Insufficiency

1993 ◽  
Vol 30 (6) ◽  
pp. 540-544
Author(s):  
G Turner ◽  
A Skinner ◽  
J S Woodhead
Keyword(s):  
Growth Hormone ◽  
Renal Insufficiency ◽  
Causative Factor ◽  
Plasma Creatinine ◽  
Tubular Dysfunction ◽  
Control Subjects ◽  
Significant Difference ◽  
Proximal Tubular ◽  
Group A ◽  
Group B

It has been reported that intrinsic renal factors could affect urinary growth hormone (UGH) measurements. We compared UGH excretion in 21 children aged 4–16 years, with various degrees of renal insufficiency, with that in 10 control subjects aged 5–13 years. We found 100- to 1000-fold elevations in UGH in children with plasma creatinine concentrations > 120 μmol/L (Group A) compared with patients with plasma creatinine concentrations < 120 μmol/L (Group B) and control subjects. UGH excretion (μU) in the three groups was as follows: group A 804–8556 (median 2649); group B 1·0–85 (median 7·5); and controls 2·6–7·3 (median 4·0). Elevated urinary β2-microglobulin levels (μg) were also observed in group A patients: 875–15 400 (median 11 637) as compared with group B, 1·0–104 (median 32) and controls, 3–18·7 (median 8·0). There was no significant difference in albumin excretion between groups A and B though six patients in group B with nephrotic syndrome (NS) excreted significantly more albumin (P < 0·05) than the other 15 patients investigated. Our data show that abnormalities of renal function have a profound effect on growth hormone excretion and we suggest proximal tubular dysfunction as the causative factor. We conclude that UGH measurements do not provide a reliable means of assessment of hypothalamo-pituitary function in patients with renal insufficiency.

scholarly journals HbA1c levels in individuals heterozygous for hemoglobin variants

2017 ◽  
Vol 63 (4) ◽  
pp. 341-346
Author(s):  
Ricardo Silva Tavares ◽  
Fábio Oliveira de Souza ◽  
Isabel Cristina Carvalho Medeiros Francescantonio ◽  
Weslley Carvalho Soares ◽  
Mauro Meira Mesquita
Keyword(s):  
Test Group ◽  
Exchange Chromatography ◽  
Group Method ◽  
Control Group ◽  
Significant Difference ◽  
Hemoglobin Variants ◽  
Different Populations ◽  
And Control ◽  
Two Populations ◽  
Hba1c Levels

Summary Objective: To evaluate the levels of glycated hemoglobin (HbA1c) in patients heterozygous for hemoglobin variants and compare the results of this test with those of a control group. Method: This was an experimental study based on the comparison of HbA1c tests in two different populations, with a test group represented by individuals heterozygous for hemoglobin variants (AS and AC) and a control group consisting of people with electrophoretic profile AA. The two populations were required to meet the following inclusion criteria: Normal levels of fasting glucose, hemoglobin, urea and triglycerides, bilirubin > 20 mg/dL and non-use of acetylsalicylic acid. 50 heterozygous subjects and 50 controls were evaluated between August 2013 and May 2014. The comparison of HbA1c levels between heterozygous individuals and control subjects was performed based on standard deviation, mean and G-Test. Results: The study assessed a test group and a control group, both with 39 adults and 11 children. The mean among heterozygous adults for HbA1c was 5.0%, while the control group showed a rate of 5.74%. Heterozygous children presented mean HbA1c at 5.11%, while the controls were at 5.78%. G-Test yielded p=0.93 for children and p=0.89 for adults. Conclusion: Our study evaluated HbA1c using ion exchange chromatography resins, and the patients heterozygous for hemoglobin variants showed no significant difference from the control group.

scholarly journals Selenium and functional constipation in children

2016 ◽  
Vol 56 (2) ◽  
pp. 111
Author(s):  
Marlina Tanjung ◽  
Supriatmo Supriatmo ◽  
Melda Deliana ◽  
Ade Rachmat Yudiyanto ◽  
Atan Baas Sinuhaji
Keyword(s):  
Abdominal Pain ◽  
Placebo Group ◽  
Side Effects ◽  
Functional Constipation ◽  
Causative Factor ◽  
Gastrointestinal Diseases ◽  
Average Frequency ◽  
Stool Consistency ◽  
Significant Difference ◽  
Selenium Treatment

Background Constipation is a common problem in children, with approximately 90 to 95% of constipation cases having functional constipation. Oxidative stress may be a causative factor in gastrointestinal diseases, alleved by intervention with antioxidants. Selenium is an essential trace element and acts as a cofactor of gluthathione peroxidase, which protects membranes from oxidative damage.Objective To determine the effect of selenium on functional constipation in children.Methods We conducted a single-blind, randomized clinical trial from November to December 2012 at the Al-Kautsar Al-Akbar Islamic Boarding School in Medan, North Sumatra. Subjects were children aged 12 to 17 years with functional constipation, diagnosed according to the ROME III criteria. Patients were randomly allocated into either the selenium group (n=57) or the placebo group (n=57). Subjects were clinically evaluated for frequency of defecation, stool consistency, severity of abdominal pain, and side effects during the 2 weeks of treatment (days 7 and 14) and 1 week after treatment had stopped (day 21).Results A total of 114 subjects were eligible to participate. The average frequency of defecation observed on day 14 was 1.5 (SD 0.75) days per defecation (P=0.0001) in the selenium group and 2.4 (0.84) days per defecation in the placebo group, a statistically significant difference (P=0.0001). There was no significant difference in frequency of defecation on the 7th day of treatment. But after day 7, there were significant differences between the groups at days 14 and 21. Normal stool consistency was found in 45 subjects (78.9%) on day 7 and in 57 subjects (100%) on day 14 of treatment in the selenium group, significantly more than those in the placebo group (P<0.05). In placebo group, normal stool consistency was found in 27 subject (47.4%) with (P=0.001) on day 7 and in 38 subject (66.7%) on day 14 of treatment (P=0.0001). On day 14, the selenium group had significantly more subjects without pain than the placebo group [47 subjects (82.5%) vs. 10 subjects (17.5%), respectively (P=0.0001)]. Severity of abdominal pain after 14th day of treatment is without pain 47 subject (82.5%) and mild pain 10 subject (17.5%) (P=0.0001). We found no side effects of selenium treatment in our subjects.Conclusion Selenium is effective in improving clinically functional constipation, in terms of increased frequency of defecation, normalization of stool consistency, and less severe abdominal pain.

scholarly journals Inhibition of Plasmin Generation in Plasma By Heparin, Low Molecular Weight Heparin and a Covalent Antithrombin-Heparin Complex (ATH)

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 4217-4217
Author(s):  
Gabriela Chang ◽  
Helen M. Atkinson ◽  
Leslie R. Berry ◽  
Anthony K.C. Chan
Keyword(s):  
Molecular Weight ◽  
Plasminogen Activator ◽  
Low Molecular Weight Heparin ◽  
Conflicts Of Interest ◽  
Area Under The Curve ◽  
Low Molecular Weight ◽  
Plasminogen Activator Inhibitor 1 ◽  
Significant Difference

Abstract Introduction: Unfractionated heparin (UFH) and low molecular weight heparin (LMWH) are widely used anticoagulants for thrombosis treatment. However, these anticoagulants have limitations such as increased bleeding, variable dose response, required frequent monitoring, and, in the case of LMWH, inability to inhibit thrombin. This has led to the development of a covalent complex of antithrombin and heparin (ATH), which has been shown to overcome many of these shortcomings. ATH has faster rates of inhibition of many coagulation factors, is able to inhibit clot-bound thrombin, and is a more effective inhibitor of both venous and arterial thrombosis in animal models. Moreover, in a rabbit thrombosis model, ATH has been shown to decrease clot mass and fibrin accretion, while the contrary was observed for UFH. From these observations, it was suggested that ATH may enhance fibrin breakdown and thus led to investigations into the effects of UFH and ATH on fibrinolysis. In vitro studies have shown that UFH enhances antithrombin inhibition of plasmin. In addition, ATH displays a slightly greater inhibition of plasmin generation and activity. Such studies were conducted in purified systems, in the absence of other plasmin inhibitors naturally present in plasma. Therefore, the aim of the present study was to compare the effects of UFH, LMWH, and ATH on plasmin generation in plasma. Methods: At 37°C tissue plasminogen activator (tPA) and soluble fibrin fragments (fib) were added to normal adult pooled platelet poor plasma supplemented with 0.35, 0.7, 1.4, or 2.1 U anti-Xa/ml UFH, LMWH, or ATH, to initiate plasmin generation (8.93nM tPA and 300µg/ml fib). At various time points, subsamples were mixed with excess plasminogen activator inhibitor 1 (PAI-1) (55.12nM) to stop further plasmin generation. The plasmin concentration at each time point was determined using a plasmin-specific chromogenic substrate and a standard curve produced from purified plasmin. Results: Comparisons of mean area under the curve (AUC) for plasmin generation displayed a significant decrease in plasmin generation in the presence of all three anticoagulants at all doses tested (p<0.05). Comparing the anticoagulants at similar doses, plasmin generation was significantly decreased in the presence of ATH (15384.66±1930.23nM/min) compared to LMWH (23892.28±3090.54nM/min) at 0.7 U/ml (p<0.05). At a dose of 1.4 U/ml, there was significantly less plasmin generated, over time, in the presence of UFH (20089.49±3022.1623nM/min) and ATH (19273.86±1805.7323nM/min) when compared to LMWH (24743.18±1265.1023nM/min) (p<0.05). There was no significant difference in plasmin inhibition between UFH and ATH at any of the doses tested. Conclusion: The present study supports previous findings that UFH and ATH can facilitate antithrombin inhibition of plasmin. It is also observed that LMWH catalyzes the inhibition of plasmin by antithrombin but possibly to a lesser extent. These findings suggest that ATH has a similar inhibitory effect on plasmin generation and activity in plasma compared to UFH, despite its overall superior anticoagulant properties. Therefore, previous in vivo observations displaying decrease in clot mass with administration of ATH was due to its enhanced anticoagulant abilities and not fibrinolysis enhancement. These findings add to our understanding of ATH mechanisms of action and aid in its development for clinical use. Disclosures No relevant conflicts of interest to declare.

scholarly journals TLR, RAGE, and HMGB1 in the Inflammatory Response in Ph(-) Myeloproliferative Neoplasm

Blood ◽  
2020 ◽  
Vol 136 (Supplement 1) ◽  
pp. 31-32
Author(s):  
Guanfang Shi ◽  
Kiron Nair ◽  
Preethi Ramachandran ◽  
Chi Chen ◽  
Ching Wong ◽  
...  
Keyword(s):  
Inflammatory Response ◽  
Real Time ◽  
Inflammatory Cytokines ◽  
Internal Control ◽  
Real Time Pcr ◽  
Mononuclear Cells ◽  
Conflicts Of Interest ◽  
Philadelphia Chromosome ◽  
Myeloproliferative Neoplasm ◽  
Significant Difference

Recent evidence of increased constitutional symptoms and inflammatory cytokines in Philadelphia chromosome negative (Ph (-)) MPN suggests that an inflammatory response is important in the pathogenesis of Ph (-) MPN. Toll-like receptors (TLR), Receptor for Advanced Glycation End products (RAGE) and High mobility group protein B1 (HMGB1) are the important pathways for the inflammatory response. All these three important pathway proteins were studied in MPN diseases in the current studies. Materials and Methods: TLR assay. TLR 2,3, 4, 7, 9 quantification was performed by immuno-staining of 1×106 mononuclear cells (peripheral blood) which were incubated with fluorescence-conjugated anti-TLR-2,3, 4, 7, 9 antibodies and assayed by flow cytometry. HMGB1assay:HMGB1 ELISA kit from Immuno-Biological Laboratories, Inc. (IBL-America) were used. The plasma samples were diluted four times with the provided sample dilution buffer, and assayed in duplicate according to the manufacturer's suggestion. RAGE (RT-PCR) Assay: Total RNA was extracted from normal control or patient mononuclear cells. Predesigned primers for RAGE, and internal control genes were ordered from Qiagen (Germantown, MD). Real-time PCR was performed using SsoAdvanced™ Universal SYBR® Green Supermix (Bio-Rad, Hercules, CA) on Bio-Rad iQ5 Multicolor Real-Time PCR Detection System. At least three house-keeping genes (ribosomal protein L4, TATA box binding protein, and tubulin-α 1b) were used as normalization controls. The expression of RAGE were compared with each internal control. Average of three was used to calculate the ratio of final patient to normal Results: Total of 97 patients with MPN were studied 1) TLR: TLR 3,7,9 was not significantly different from controls. But TLR 2 was significantly increased in both PV, as well as in the MPN group when PV, ET and MF were grouped together as MPN (Fig A). TLR 4 was not significantly increased in PV, ET, MF individually but was found to be significantly increased than the controls, when they are grouped together as MPN (Fig B). 2) RAGE: No significant difference was found between ET, PV, MF individually or when they were grouped together as MPN than the controls (Fig C). 3) HMGB1: No significant difference was seen between ET, PV, MF or when they were grouped as MPN (Fig D). Conclusion: Current study suggests that TLR pathway especially TLR2, and to a lesser extent TLR4 are the important pathways for inflammatory response with increased inflammatory cytokines in MPN, while HMGB1 and RAGE pathways were not different from controls. Figure Disclosures No relevant conflicts of interest to declare.

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