The Comparative Immunotropic Activity of Carrageenan, Chitosan and Their Complexes

The immunotropic activity of polyelectrolyte complexes (PEC) of κ-carrageenan (κ-CGN) and chitosan (CH) of various compositions was assessed in comparison with the initial polysaccharides in comparable doses. For this, two soluble forms of PEC, with an excess of CH (CH:CGN mass ratios of 10:1) and with an excess of CGN (CH: CGN mass ratios of 1:10) were prepared. The ability of PEC to scavenge NO depended on the content of the κ-CGN in the PEC. The ability of the PEC to induce the synthesis of pro-inflammatory (tumor necrosis factor-α (TNF-α)) and anti-inflammatory (interleukine-10 (IL-10)) cytokines in peripheral blood mononuclear cell was determined by the activity of the initial κ-CGN, regardless of their composition. The anti-inflammatory activity of PEC and the initial compounds was studied using test of histamine-, concanavalin A-, and sheep erythrocyte immunization-induced inflammation in mice. The highest activity of PEC, as well as the initial polysaccharides κ-CGN and CH, was observed in a histamine-induced exudative inflammation, directly related to the activation of phagocytic cells, i.e., macrophages and neutrophils.

Characterization of sheep erythrocyte glycosphingolipids recognized by human anti-Forssman antibodies

The FORS histo-blood group system is the most recently discovered carbohydrate-based human blood group system. FORS is a rare blood group system, and most individuals have naturally occurring anti-FORS1 antibodies in plasma. Screening for anti-FORS1 antibodies is often done by hemagglutination assays using FORS1-expressing sheep erythrocytes, since FORS1-positive human erythrocytes are most often not available. Here, we have characterized the non-acid glycosphingolipids from sheep erythrocytes and isolated subfractions, with mass spectrometry, binding of antibodies and lectins, and by enzymatic hydrolysis. This demonstrated the presence of Forssman and Galili pentaosylceramides, and a Galili heptaosylceramide. Two complex glycosphingolipids recognized by human anti-FORS1 antibodies were characterized as a Forssman neolacto hybrid hexaosylceramide (GalNAcα3GalNAcβ3Galβ4GlcNAcβ3Galβ4Glcβ1Cer) and a Forssman Galili hybrid heptaosylceramide (GalNAcα3GalNAcβ3Galα3Galβ4GlcNAcβ3Galβ4Glcβ1Cer). These are novel glycosphingolipid structures, and to our knowledge, the first case of an elongated Galili antigen. Thus, the anti-Forssman antibodies in human serum bind not only to the classical Forssman pentaosylceramide (GalNAcα3GalNAcβ3Galα4Galβ4Glcβ1Cer), but also when the GalNAcα3GalNAcβ3 sequence is presented on a neolacto core chain and even on a Galili carbohydrate sequence.

THE EFFECTS OF DIFFERENT STORING TEMPERATURES ON THE ACTIVITY OF SHEEP BLOOD AND PLASMA GLUTATHIONE PEROXIDASE

The aim of the study was to determine the stability of the activity ofglutathione peroxidases 1 and 3 during storage at +40C and -180C. Blood sampleswere taken from eight sheep and the activity of the enzyme was determined in theplasma (GPx3) and erythrocytes (GPx1) on the first, third, fifth and seventh day insamples stored at +40C and after one and three months in samples stored at -180C.GPx3 activity decreased significantly during storage at both temperatures, whileGPx1 remained steady even after three months of storage at -180C. Obtained resultsindicate that GPx3 activity has to be determined in fresh sheep plasma samples,while the activity of sheep erythrocyte GPx1 can be determined even after 3 monthsof storage at -180C.

In VitroProtective Effect of Phikud Navakot Extraction on Erythrocyte

Phikud Navakot (PN), Thai herbal remedy in National List of Essential Medicines, has been claimed to reduce many cardiovascular symptoms especially dizziness and fainting. Apart from blood supply, erythrocyte morphology, in both shape and size, is one of the main consideration factors in cardiovascular diseases and may be affected by vascular oxidative stress. However, little is known about antioxidative property of PN on erythrocyte to preserve red blood cell integrity. In this study, 1,000 μM hydrogen peroxide-induced oxidative stress was conducted on sheep erythrocyte. Three doses of PN (1, 0.5, and 0.25 mg/mL) and 10 μM of ascorbic acid were compared. The released hemoglobin absorbance was measured to demonstrate hemolysis. Electron microscopic and immunohistochemical studies were also performed to characterize dysmorphic erythrocyte and osmotic ability in relation to aquaporin- (AQP-) 1 expression, respectively. The results revealed that all doses of PN and ascorbic acid decreased the severity of dysmorphic erythrocyte, particularly echinocyte, acanthocyte, knizocyte, codocyte, clumping, and other malformations. However, the most effective was 0.5 mg/mL PN dosage. In addition, hydrostatic pressure may be increased in dysmorphic erythrocyte in association with AQP-1 upregulation. Our results demonstrated that PN composes antioxidative effect to maintain the integrity and osmotic ability on sheep erythrocyte.

T- Cell Depleted Peripheral Blood Stem Cell (TCD-PBSC) Transplants Secure Consistent Engraftment with Low Risk of Acute or Chronic Gvhd and Favorable Disease Free Survival (DFS) and Overall Survival (OS) for Pediatric Patients (<21 years) with AML in CR1 or CR2 or MDS Including tMDS/AML

Higher cGVHD, TRM and treatment failure following sibling donor unmanipulated allogeneic PBSCT compared to BMT in children and adolescents has been reported by the IBMTR (Eapen M et al, J Clin Oncol, 2004). To determine the effect of T cell depletion (TCD) of PBSC on outcome for pediatric patients with AML or MDS, we reviewed our results with 49 patients (pts) with AML-1CR (n=18), AML-CR2 (n=11), MDS (n=10) and tAML/MDS (n=8/2) following allogeneic TCD-PBSCT transplanted between June 2003 and January 2015. Median age in this cohort was 13.9yrs (range 1.9 - 20.4) with 27 males and 19pts CMV seropositive at time of PBSCT. Donors were related HLA identical (14siblings, 1uncle), unrelated HLA 10/10 matched (n=14) or unrelated HLA mismatched (n=20: 9/10 n=12: 8/10 n=8). Cytoreduction consisted of a HFTBI containing regimen (n=14) with thiotepa and cyclophosphamide (n=12) or thiotepa and fludarabine (n=2). Alternatively, 35 pts received chemotherapy alone: BU/MEL/FLU (n=31) or other (n=4). ATG provided graft rejection prophylaxis for 43 pts either pre (n=37) or post (n=6) infusion. Six young pts with sibling donors did not receive ATG. No pt received additional GVHD prophylaxis. PBSC were CD34+ selected using ISOLEX 300i Magnetic Cell Selection System, followed by sheep erythrocyte rosetting (n=25) or CD34+ selected with Miltenyi CliniMACS device (n=24). The median CD34+ cells/Kg was 10.5x106 (range 3.3 - 33.4x106) and CD3+ cells/kg was 2.4x103 (range 0 - 29x103). All pts engrafted neutrophils and platelets at a median of 11 days (range 9 - 14 days) and 26 days (range 13 - 182 days), respectively. 42pts had platelet recovery by day 40. One patient who had late graft failure (GF) at 3.5 mos engrafted with a subsequent unmanipulated PBSCT. Seven pts developed Grade II or III aGVHD of skin or GI. No pt had Gr IV aGVHD. Six pts responded to treatment. Only 1pt developed limited cGVHD of the skin. No pt who was CMV seronegative developed CMV viremia. 7 of 19 CMV seropositive pts developed CMV viremia. Nine pts received rituximab for EBV viremia. No pt died of CMV disease or EBV-LPD. Five pts received defibrotide for treatment of VOD. Table 1.N1o GF2o GFaGVHD (Gr 2-3)cGVHD (limited)CMV-CTLEBV-CTLRelapseAlive in CR p2nd SCT for GF/rel3 yrs DFS3 yr OSEntire Cohort4901(2%)6(12%)1 (2%)1(2%)1(2%)11(22%)5(4rel,1GF)72.3%87.7%AML-CR11800210043(3rel)75.6%94.4AML-CR2110020114056.0%66.7MDS1001000022(1GF,1rel)78.8%100%tAML/MDS100020001080.0%88.9% With a median follow-up of 36.3 mos the DFS and OS at for the entire cohort at 3 yrs is 72.3% and 87.7%, respectively. Although the DFS for pts with AML-CR1 is 75.6%, 3 of 4 pts are in CR2 post a second BMT for OS of 94.4%. The 4 pts with AML-CR2 who relapsed died of disease. In this series of 49 pts, 6pts died of disease, 1 pt died of aGVHD and 1 pt (Down Syndrome) died of MOF (VOD). Thus, in young pts (< 21 yrs) TCD-PBSCT is associated with low risk of acute and chronic disease, no increment in relapse and a highly favorable DFS and OS. The data support the inclusion of pediatric pts in BMT CTN 1301 trial (NCT02345850): a Randomized, Multi-Center, Phase III Trial of Calcineurin Inhibitor-Free Interventions for Prevention of Graft-versus-Host Disease. Disclosures Kernan: Gentium S.p.A.: Research Funding. Hasan:Atara Biotherapeutics: Research Funding. O'Reilly:Atara Biotherapeutics: Research Funding.