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The aim of the present study was to evaluate the circulating T regulatory cells (Tregs) in patients undergoing extracorporeal photopheresis (ECP) for the prevention of chronic graft-versus-host disease (GvHD) and to search for any correlation between Tregs counts and chronic GvHD occurrence. Among n = 12 patients with complete longitudinal data, the median cumulative values of absolute peripheral Tregs counts were 21.64 and 63.49 cells/µL for patients who developed chronic GvHD and those who did not develop it, respectively (p = 0.05). The analysis of the median absolute counts of peripheral HLA-DR + Tregs provided similar results, showing that 20% (1 out of 5) and 100% (7 out of 7) of patients with HLA-DR + Tregs values of > 5 cells/µL were in the GvHD and non-GvHD groups, respectively (p = 0.01). In conclusion, the present results support the involvement of Tregs in the prevention of chronic GvHD in patients receiving ECP and suggest Tregs count as a potential biomarker of ECP effectiveness. Future strategies are needed to enhance Tregs expansion and/or activity in conjunction with ECP for an effective chronic GvHD prevention.

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The production of bleached cellulose pulps from elephant grass (Miscanthus sinensis) via a two-stage soda pulping process and a TCF bleaching sequence is evaluated in this work. The impregnation rapid steam pulping process (IRSP) involves impregnating of the lignocellulosic material with the pulping liquor, withdrawing the excess liquor and rapidly steaming the impregnated material at 180–200°C for a short time. In this paper the process variables and their effect on the kappa number, yield and viscosity of the unbleached pulps are discussed. Bleaching by an ozone-based TCF sequence was tested, and the papermaking properties of the bleached pulp were determined. A kappa number of 19 was obtained by impregnating at an alkali charge of 30 + 0.1% anthraquinone carboxylic acid (AQCA) and pulping at 180°C for only 15 min. Kappa was reduced to 16 by extending pulping time to 26 min. The alkali consumption during impregnation and pulping was 10.2 g NaOH/100 g of dry Miscanthus. Screened pulp yield, visc...

This paper explores the production of carboxymethylcellulose (CMC) fromseveral bleached cellulose pulps obtained from non-wood species. The chemicalcomposition (α-cellulose, hemicellulose and lignin content), the degreeofswelling, viscosity, solubility in concentrated NaOH and crystallinity ofsoda/AQ cellulose pulps from abaca, jute, sisal, linen and Miscanthussinensis were determined. The pulps were carboxymethylated by one andtwo successive reaction steps in aqueous medium under identical conditions. Thedegree of substitution (DS) of CMC was found to be dependent upon the source ofthe cellulose pulp, but generally it was close to 1 with one etherificationtreatment and around 2 after the second. The molar mass of CMC was found to bedependent on the initial intrinsic viscosity of the cellulose pulp. The weightaverage molar mass of our CMCs ranged from 1.5 × 105 to2.8 × 105. Increasing the DS up to 2 improved the CMC solubility, butviscosity slightly decreased due to a slight degradation of the polymer.

This paper concerns the rheological behavior of carboxymethylcelulloses(CMC) derived after one and two successive steps from different non-woodbleached cellulose pulps. CMC rheological characterization was achieved in0.1M NaCl solution, as a function of polymer concentration. Theevidence of a critical concentration (C* < 1 g/L) is discussedfromsteady shear and dynamic experiments. Rheological properties of the CMC werefound to depend on the cellulose source reactivity and on their degree ofsubstitution (DS). Higher molecular weight of initial cellulose was accompaniedby higher apparent intrinsic viscosity of the CMC produced. Depending on theCMCconcentration and on the degree of etherification, the system behaves as asolution or as a gel. In the case of abaca CMC sample, it is shown that afteronly one step of chemical modification and above a polymer concentration of20 g/L, the system behaves as a gel. The gel behavior was studied asafunction of temperature. In the temperature range from 25 to 45°C, the rheological behavior was found to remain almostconstant due to the existence of dispersed swollen aggregates. This unusualcharacteristic represents an advantage for applications such as oil recovery inthe petroleum industry, where viscosity of the recovered fluid should not diminishwith temperature.

An independent clinical assessment was compared with flow cytometry (FCM) and cytomorphology results obtained on 227 cerebrospinal fluids investigated for hematologic malignancy, in a retrospective longitudinal study with a median observation time of 11 months. A combined method assessment (CMA), defining "positive" a sample if at least one method gave "positive" results, was also tested. Eleven out of 55 screening samples and 53 out of 166 follow-up samples resulted positive at clinical evaluation. FCM and CM were concordant with positive clinical assessment in 68.5% and 51.5% of cases, respectively. According to CMA, 10.5% of samples (resulting false negative by either FCM or cytomorphology) were rescued as true positive. FCM retained significantly higher accuracy than cytomorphology (p = 0.0065) and 100% sensitivity when at least 220 leukocytes were acquired. CMA accuracy was higher than FCM accuracy and significantly higher than cytomorphology accuracy in the analysis of all samples (p < 0.0001), samples from mature B/T cell neoplasms (p = 0.0021), and samples drawn after intrathecal treatment (p = 0.0001). When acquiring ≥220 leukocytes, FCM accuracy was poor, and combining cytomorphology added statistically significant diagnostic advantage (p = 0.0043). Although FCM is the best diagnostic tool for evaluating CSF, morphology seems helpful especially when clinically positive follow-up samples are nearly acellular.

Flow cytometry and cytomorphology results on 92 body cavity fluids [61 effusions and 31 bronchoalveolar lavage fluids (BALF)] from hematologic malignancy were compared with retrospective clinical outcome. We observed double true positive/negative results in 67 cases (73%), and double false negative results in 2 cases (2%). Immunophenotyping accounted for true positive/negative results in 22 out of 23 mismatched cases (25%), and retained significantly higher accuracy than that of cytomorphology especially in effusions and differentiated lymphoma. In BALF analysis, immunophenotyping and cytomorphology sensitivity was 75% and 0%, respectively. Flow cytometry retains the highest accuracy in detecting neoplastic cells in body cavity fluids.

Introduction: Flow cytometry is well known to detect malignant cells in peripheral blood and bone marrow of patients with hematologic malignancies. However, its role in evaluating non-hematic body fluid contamination by tumor cells is largely unexplored. Patients and Methods: Data detected by flow cytometry in non-hematic body fluid samples drawn between 2002 and 2007 from patients with hematologic neoplasms were retrospectively compared with morphological findings obtained from cytospin slides. Immunophenotyping was carried out by using disease-specific multicolor panels of quadruple monoclonal antibodies, conjugated with the fluorochromes FITC, PE, PerCP, and APC, respectively. Acquisition of information on 1x104 to 1x105 stained cells depending on the whole sample cellularity was assessed on a dual-laser FACSCalibur flow cytometer using the CellQUEST software (Becton Dickinson, San José, CA, USA). Results: Fourty-five samples (bronchoalveolar fluid, n=5; ascites, n=2; hydrocele, n=2; pleural effusion, n=8; and cerebrospinal fluid, n=28) from 32 patients were available for comparison. Diagnoses were as follows: chronic myelomonocitic leukemia (n=1), acute promyelocytic leukaemia (n=2), acute myelomonocytic leukaemia (n=1), B-chronic lymphocitic leukemia (n=4), follicular lymphoma (n=1), acute lymphoblastic leukaemia (n=6), lymphoblastic lymphoma (n=1), high grade non-Hodgkin’s lymphoma (NHL), Burkitt-like (n=3), diffuse large B-cell NHL (n=6), peripheral T-cell NHL (n=3), and NHL, unspecified (n=4). Flow cytometry detected neoplastic cells in 24 cases. Of these cases, only 17 were positive also by morphology. In 7 cases, in which tumor cells were detected by flow cytometry but not by morphology, clinical data confirmed the presence of the disease. Flow cytometry did not show neoplastic cells in 21 cases. Of these cases, only 18 were negative also by morphology. In the remaining 3, the suggestion of diffuse large B-cell NHL contamination by morphology was not confirmed by flow cytometry, demonstrating T-reactive lymphocytes that were clearly negative for disease-specific markers. Conclusions: Our data suggest that flow cytometry is a useful tool complementary to morphology for the screening of non-hematic body fluid contamination in patients with hematologic neoplasms.

Abstract 5018 Flow cytometry (FCM) and cytomorphology (CM) evaluation of hematologic malignancy in hypercellular body cavity fluids (BCF) needs clinical validation and standardized approaches for definitive sample assessment. A total of 1,011 hypercellular BCF analyzed by FCM between 2002 and 2009 were retrospectively examined. As a prerequisite for inclusion in the study, each of the following was needed: suspected or known hematologic malignancy at withdrawal, CM performed on the same sample, and availability of follow-up data for reliable retrospective clinical outcome (RCO). Ninety-two BCF [61 effusions, 31 bronchoalveolar lavage fluids (BALF)] were selected for the analysis. Results obtained by the two methods were matched with RCO. A combined method assessment (CMA) was also tested, accounting as negative those samples not assessed as frankly positive by at least one method. Thirty-six percent of BCF resulted RCO-positive. By applying each method to RCO, concordant results between FCM and CM were observed in 69 cases (75%). Fifty-six cases (61%) were double true negative (TN), 11 cases (12%) were double true positive (TP), and 2 cases (2%) were double false negative (FN). The latter were BALF samples from patients with primary pulmonary lymphomatoid granulomathosis in which FCM revealed a prominent inflammatory background of CD3+ T-cells; clonality could not be assessed due to the small number of B-cells. Mismatched results between FCM and CM were observed in 23 cases (25%). FCM gave TP results in 19 cases (20.5%) with FN or uncertain CM, TN results in 3 cases (3%) with uncertain CM, and FN results in 1 case (1%) with TP CM. This was a pleural effusion in which, in the presence of anaplastic cell features, primary effusion lymphoma was definitively diagnosed by immunocytochemical staining for ORF73/latent nuclear antigen-1 of HHV-8 on B-cell marker negative neoplastic cells. By applying the CMA to RCO, 59 TN results (64%), 31 TP results (33.5%) and 2 (double) FN results (2%) were obtained. Overall, 100% specificity and positive predictive value were detected for each method. FCM/CMA sensitivity was significantly higher than CM sensitivity (p In conclusion, clinical follow-up demonstrates FCM to retain significantly higher accuracy than CM in detecting hematologic malignancy contamination in both effusions and BALF, especially in the subset of differentiated lymphomas. Combining methods improves diagnostic performance almost completely by means of FCM. Disclosures No relevant conflicts of interest to declare.

Cellulose pulps were prepared from ground branch wood from pine and poplar and ground wheat straw by a fast soda/anthraquinone pulping. This process used high pulping temperatures and short residence times. Various combinations of temperature and time were used so that the H factor for pulps from pine and poplar was a constant 2700 while for pulps from wheat straw it was 270. The chemical composition of the resulting cellulose pulps was characterized and the possibility of synthesizing carboxymethyl cellulose from them was evaluated. The degree of substitution of our CMCs was found to be dependent upon the source of the lignocellullosic material, but independent of the pulping conditions for a specific substrate, (0.75 for pine, 0.82 for poplar and 0.89 for wheat straw). The rheological properties, the intrinsic viscosity and the molecular weight of the CMCs were found to be dependent on the pulping conditions for each lignocellulosic tested. The molecular weights of our CMC samples were 219,000 ...