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The theory of universal emotions suggests that certain emotions such as fear, anger, disgust, sadness, surprise and happiness can be encountered cross-culturally. These emotions are expressed using specific facial movements that enable human communication. More recently, theoretical and empirical models have been used to propose that universal emotions could be expressed via discretely different facial movements in different cultures due to the non-convergent social evolution that takes place in different geographical areas. This has prompted the consideration that own-culture emotional faces have distinct evolutionary important sociobiological value and can be processed automatically, and without conscious awareness. In this paper, we tested this hypothesis using backward masking. We showed, in two different experiments per country of origin, to participants in Britain, Chile, New Zealand and Singapore, backward masked own and other-culture emotional faces. We assessed detection and recognition performance, and self-reports for emotionality and familiarity. We presented thorough cross-cultural experimental evidence that when using Bayesian assessment of non-parametric receiver operating characteristics and hit-versus-miss detection and recognition response analyses, masked faces showing own cultural dialects of emotion were rated higher for emotionality and familiarity compared to other-culture emotional faces and that this effect involved conscious awareness.

Commercial aluminum foils were coated by graphene oxide, and its functionalized derivatives and the corrosion performance of the coated specimens were examined in acidic conditions (lithium perchlorate and sulfuric acid). Electrochemical experiments have shown that all graphene oxide-coated specimens provided up to 96% corrosion inhibition efficiency with a corresponding lower corrosion rate compared to the bare aluminum foil. Our results clearly show that graphene-related materials offer viable alternatives for the protection of aluminum, and this opens up a number of possibilities for its use in a number of commercial applications.

Modern and contemporary art materials are generally prone to irreversible colour changes upon exposure to light and oxidizing agents. Graphene can be produced in thin large sheets, blocks ultraviolet light, and is impermeable to oxygen, moisture and corrosive agents; therefore, it has the potential to be used as a transparent layer for the protection of art objects in museums, during storage and transportation. Here we show that a single-layer or multilayer graphene veil, produced by chemical vapour deposition, can be deposited over artworks to protect them efficiently against colour fading, with a protection factor of up to 70%. We also show that this process is reversible since the graphene protective layer can be removed using a soft rubber eraser without causing any damage to the artwork. We have also explored a complementary contactless graphene-based route for colour protection that is based on the deposition of graphene on picture framing glass for use when the direct application of graphene is not feasible due to surface roughness or artwork fragility. Overall, the present results are a proof of concept of the potential use of graphene as an effective and removable protective advanced material to prevent colour fading in artworks.

Graphene and other two-dimensional materials (2DMs) have been shown to be promising candidates for the development of flexible and highly-sensitive strain sensors. However, the successful implementation of 2DMs in practical applications is slowed down by complex processing and still low sensitivity. Here, we report on a novel development of strain sensors based on Marangoni self-assemblies of graphene and of its hybrids with other 2DMs that can both withstand very large deformation and exhibit highly sensitive piezoresistive behaviour. By exploiting the Marangoni effect, reference films of self-assembled reduced graphene oxide (RGO) are first optimized, and the electromechanical behaviour has been assessed after deposition onto different elastomers demonstrating the potential of producing strain sensors suitable for different fields of application. Hybrid networks have been then prepared by adding hexagonal boron nitride (hBN) and fluorinated graphene (FGr) to the RGO dispersion. The hybrid integration of 2D materials is demonstrated to become a potential solution to increase substantially the sensitivity of the produced resistive strain sensors without compromising the mechanical integrity of the film. In fact, for large quasi-static deformations, a range of gauge factor values up to 2000 were demonstrated, while retaining a stable performance under cyclic deformations.

Graphene-related materials (GRMs) are subject to intensive investigations and considerable progress has been made in recent years in terms of safety assessment. However, limited information is available concerning the hazard potential of GRM-containing products such as graphene-reinforced composites. In the present study, we conducted a comprehensive investigation of the potential biological effects of particles released through an abrasion process from reduced graphene oxide (rGO)-reinforced composites of polyamide 6 (PA6), a widely used engineered thermoplastic polymer, in comparison to as-produced rGO. First, a panel of well-established in vitro models, representative of the immune system and possible target organs such as the lungs, the gut, and the skin, was applied. Limited responses to PA6-rGO exposure were found in the different in vitro models. Only as-produced rGO induced substantial adverse effects, in particular in macrophages. Since inhalation of airborne materials is a key occupational concern, we then sought to test whether the in vitro responses noted for these materials would translate into adverse effects in vivo. To this end, the response at 1, 7 and 28 days after a single pulmonary exposure was evaluated in mice. In agreement with the in vitro data, PA6-rGO induced a modest and transient pulmonary inflammation, resolved by day 28. In contrast, rGO induced a longer-lasting, albeit moderate inflammation that did not lead to tissue remodeling within 28 days. Taken together, the present study suggests a negligible impact on human health under acute exposure conditions of GRM fillers such as rGO when released from composites at doses expected at the workplace., This work was supported by the European Union (EU) 8th Framework Program for Research and Technological Development, Graphene Flagship project (H2020-FET- GrapheneCore2 - #785219 and H2020-FET- GrapheneCore3 - #881603). Part of this work was supported by the University of Trieste, the Agence Nationale de la Recherche (ANR) through the LabEx project Chemistry of Complex Systems (ANR-10-LABX- 0026_CSC), the Maria de Maeztu Units of Excellence Program from the Spanish State Research Agency – Grant No. MDM-2017-0720 and the Swiss National Science Foundation (Grant 310030_169207).

The theory of universal emotions suggests that certain emotions such as fear, anger, disgust, sadness, surprise and happiness can be encountered cross-culturally. These emotions are expressed using specific facial movements that enable human communication. More recently, theoretical and empirical models have been used to propose that universal emotions could be expressed via discretely different facial movements in different cultures due to the non-convergent social evolution that takes place in different geographical areas. This has prompted the consideration that own-culture emotional faces have distinct evolutionary important sociobiological value and can be processed automatically, and without conscious awareness. In this paper, we tested this hypothesis using backward masking. We showed, in two different experiments per country of origin, to participants in Britain, Chile, New Zealand and Singapore, backward masked own and other-culture emotional faces. We assessed detection and recognition performance, and self-reports for emotionality and familiarity. We presented thorough cross-cultural experimental evidence that when using Bayesian assessment of non-parametric receiver operating characteristics and hit-versus-miss detection and recognition response analyses, masked faces showing own cultural dialects of emotion were rated higher for emotionality and familiarity compared to other-culture emotional faces and that this effect involved conscious awareness.

Aerogels have attracted significant attention recently due to their ultra-light weight porous structure, mechanical robustness, high electrical conductivity, facile scalability and their use as gas and oil absorbers. Herein, we examine the multi-functional properties of hybrid aerogels consisting of reduced graphene oxide (rGO) integrated with hexagonal boron nitride (hBN) platelets. Using a freeze-drying approach, hybrid aerogels are fabricated by simple mixing with various volume fractions of hBN and rGO up to 0.5/0.5 ratio. The fabrication method is simple, cost effective, scalable and can be extended to other 2D materials combinations. The hybrid rGO/hBN aerogels (HAs) are mechanically robust and highly compressible with mechanical properties similar to those of the pure rGO aerogel. We show that the presence of hBN in the HAs enhances the gas absorption capacities of formaldehyde and water vapour up to ~ 7 and > 8 times, respectively, as compared to pure rGO aerogel. Moreover, the samples show good recoverability, making them highly efficient materials for gas absorption applications and for the protection of artefacts such as paintings in storage facilities. Finally, even in the presence of large quantity of insulating hBN, the HAs are electrically conductive, extending the potential application spectrum of the proposed hybrids to the field of electro-thermal actuators. The work proposed here paves the way for the design and production of novel 2D materials combinations with tailored multi-functionalities suited for a large variety of modern applications.

Introduction In B-acute lymphoblastic leukemia (B-ALL), the identification of cytogenetic prognostic factors is important for stratifying patients into risk groups and tailoring treatment accordingly. The purpose of this study was to propose flow cytometric (FCM) scoring systems (SSs) for predicting t(12;21)(p13;q22), t(9;22)(q34;q11), t(11q23), and t(1;19)(q23;p13.3) translocations. Methods We analyzed retrospectively the FCM immunophenotype of 377 patients with B-ALL with regard to the major cytogenetic findings revealed by interphase fluorescence in situ hybridization (i-FISH). Comparing descriptive data on the expression of each antigen and performing receiver operating characteristic (ROC) analysis, we identified the most reliable predictive markers for each translocation and sought to establish a specific SS for each translocation, based on specific antibody panels. Results CD27, CD9, CD66c, CD10, CD25, and CD34 were employed for the prediction of t(12;21), CD25, CD38, CD34, and CD66c for t(9;22), NG2, CD10, CD15, CD34, and CD20 for t(11q23), and CD34, cμ, CD123, and CD66c for t(1;19). The sensitivity and specificity, respectively, of each predictive score were 89.29% and 96.15% for t(12;21), 75.00% and 88.19% for t(9;22), 84.21% and 99.04% for t(11q23), and 85.71% and 92.71% for t(1;19). Conclusion Four highly specific and significantly sensitive FCM-obtained SSs are proposed for the prediction of the four major translocations observed in patients with B-ALL. Prospective evaluation of the proposed SSs could lead to a better targeted cytogenetic investigation and therefore to more cost-effective laboratory practice.

International audience; The development of three-dimensional (3D) porous graphitic structures is of great interest for electrochemical sensing applications as they can support fast charge transfer and mass transport through their extended, large surface area networks. In this work, we present the facile fabrication of conductive and porous graphitic electrodes by direct laser writing techniques. Irradiation of commercial polyimide sheets (Kapton tape) was performed using a low-cost laser engraving machine with visible excitation wavelength (405 nm) at low power (500 mW), leading to formation of 3D laser-induced graphene (LIG) structures. Systematic correlation between applied laser dwell time per pixel ("dwell time") and morphological/structural properties of fabricated electrodes showed that conductive and highly 3D porous structures with spectral signatures of nanocrystalline graphitic carbon materials were obtained at laser dwell times between 20 and 110 ms/pix, with graphenelike carbon produced at 50 ms/pix dwell time, with comparable properties to LIG obtained with high cost CO 2 lasers. Electrochemical characterization with inner and outer sphere mediators showed fast electron transfer rates, comparable to previously reported 2D/3D graphene-based materials and other graphitic carbon electrodes. This work opens the way to the facile fabrication of low-cost, disposable electrochemical sensor platforms for decentralized assays.

The prognostic significance of the ETV6/RUNX1-fusion and of the accompanying aberrations is disputable; whether co-existing sub-clones are responsible for delayed MRD-clearance and thus, moderate outcome, remains to be clarified. We studied, in a paediatric cohort of 119 B-ALLs, the relation between the ETV6/RUNX1 aberration and the co-existing subclones with (a) presenting clinical/biological features, (b) early response to treatment(MRD) and (c) long-term outcome over a 12-year period. Patients were homogeneously treated according to BFM-based-protocols. 27/119 patients (22.7%) were ETV6/RUNX1-positive; 19/27 (70.4%) harbored additional genetic abnormalities while 9/19 (33.3%) presented with clonal heterogeneity. The most common abnormalities were del12p13 (37%), 3-6×21q22 (22.2%), del9p21 (18.5%) and 2-3xETV6/RUNX1 (18.5%). MRD d15 -positivity (≥10 −3 ) was detected in 44% of the cohort; the corresponding MRD among patients carrying subclones rises to 88.9%. Common features of all relapses were sub-clonal diversity, FCM-MRD d15 -positivity and additional del(9p21) while there were no censored relapses among ETV6/RUNX1-positive patients with sole translocation and absence of additional aberrations, within a median follow-up time of 90 months. In our study, the presence of clonal heterogeneity and impaired FCM-MRD clearance among ETV6/RUNX1-positive patients, ultimately influenced prognosis. Longer follow-up is needed in order to further validate these initial results.

AimsThe causes and diagnosis of ‘double-negative’ (CD3+CD4−CD8−) T-cell lymphocytosis are not well studied. We aimed to define the causes of double-negative T-cell lymphocytosis in children and adults, and to identify simple clinical and laboratory features that would help to differentiate between the underlying conditions.MethodsWe collected clinical and laboratory data on 10 children and 30 adults with significantly increased peripheral-blood double-negative T-cells (>10% of total lymphocytes). We identified conditions associated with double-negative T-lymphocytosis with flow cytometry, peripheral-blood morphology and T-cell receptor-gene rearrangement studies. Patients were assigned to diagnostic categories on the basis of these test results.Results and conclusionsThe causes of double-negative T-cell lymphocytosis in children were autoimmune lymphoproliferative syndrome (ALPS) and reactive γ/δ Τ-lymphocytosis. T-cell large granular lymphocyte (T-LGL) leukaemia, reactive γ/δ T-lymphocytosis and hepatosplenic T-cell lymphoma (HSTL) were the the most common disorders underlying double-negative T-cell lymphocytosis in adults. Less common causes included hypereosinophilic syndrome, peripheral T-cell lymphoma, ALPS and monoclonal, double-negative T-lymphocytosis of uncertain significance. CD5/CD7/Vδ2 expression and absolute double-negative lymphocyte count (9/L) were useful discriminators for distinguishing patients with reactive γ/δ T-lymphocytosis from those with γ/δ lymphoproliferative disorders. Differentiating between γ/δ T-LGL and HSTL can be difficult. Expression of CD57 and cellular morphology (pale cytoplasm with distinct granules) would support a diagnosis of γ/δ T-LGL.

Graphene wetting by polymers is a critical issue to both the success of polymer-aided transfer of large size sheets onto specific substrates and to the development of well performing nanocomposites. Here we show for the first time that high temperature contact angle measurements can be performed to investigate the wettability of CVD graphene by molten polymers. In particular, poly(methyl methacrylate), a widely used polymer support for CVD graphene transfer, has been adopted herein for this proof-of-concept study and the values of contact angle and work of adhesion have been provided in the temperature range 170–200 °C.

The association between mature-B phenotype and MLL abnormalities in acute lymphoblastic leukemia (ALL) is a very unusual finding; only 14 pediatric cases have been reported so far. We describe the clinical and biological characteristics and outcome of five pediatric cases of newly diagnosed B lineage ALL with MLL abnormalities and mature immunophenotype based on light chain restriction and surface Ig expression. Blasts showed variable expression of CD10/CD34/TdT. MLL abnormalities with no MYC involvement were detected in all patients by G-banding, FISH, and/or RT-PCR. Three patients were treated according to Interfant protocol, one to ALLIC-09, and one received B-NHL-BFM-2004. All patients achieved complete remission and three of them relapsed. Despite the small cohort size, it could be postulated that B lineage ALL with MLL abnormalities and mature phenotype is a distinct entity that differs both from the typical Pro B ALL observed in infants and mature B-ALL with high MYC expression.

The work investigates some of the differences and similarities between fully-premixed and stratified lean propane flames stabilized in an axisymmetric bluff-body swirl burner configuration. In this arrangement a double-cavity propane–air premixer, formed along three concentric disks, supplies the afterbody disk recirculation region with a regulated radial equivalence ratio gradient. Flame stabilization can be established at a range of co-flow swirl intensities under, either uniformly premixed or stratified inlet mixture conditions. Measurements of turbulent velocities, temperatures, chemiluminescence imaging of OH* and CH* and gas analysis provided information for lean and close to blow-off flame configurations. Supporting Large Eddy Simulations were undertaken with the Thickened Flame Model and a nine-step mechanism for propane combustion to complement the experiments. The combined methodology helped to elucidate some of the parametric characteristics of the diverse flame topologies obtained in the above burner. The performance of the adopted turbulent chemistry closure and the reduced chemical scheme in addressing the effect of mixture heterogeneities on the flame anchoring and structure of the studied configurations is also evaluated. The interaction of the toroidal flame fronts with the recirculation stabilization region, the impact of heat release on the development of the axisymmetric wakes and the flame front disposition at ultra-lean and close to blow-off operation are presented and discussed for the different fuel–air inlet mixture profiles.

The innate immune system is the first line of defense and plays a key role in thyroid cancer development. The role of the tumor-infiltrating natural killer (NK) cells is becoming increasingly important in research and potential cancer therapies. NK cell subpopulations, CD3(-)CD16(+)CD56(dim) and CD3(-)CD16(-)CD56(bright), demonstrate a significant role in the tumor immuno-surveillance process.We investigated the distribution of CD3(-)CD16(+)CD56(dim) and CD3(-)CD16(-)CD56(bright) NK subpopulations in tissue and blood samples from patients with papillary thyroid cancer (PTC) and nodular goiter (NG). Twenty-eight patients with PTC, 13 patients with NG, and 50 healthy donors were included in the study. Tissue and blood samples from all patients and blood samples from healthy donors were analyzed for CD3(-)CD16(+)CD56(dim) and CD3(-)CD16(-)CD56(bright) NK cells by flow cytometry.A significant predominance of CD3(-)CD16(+)CD56(dim) cells compared to CD3(-)CD16(-)CD56(bright) NK cells was found in blood samples in all groups (p0.0001 in PTC, NG, and healthy donors). Increased infiltration by CD3(-)CD16(-)CD56(bright) NK cells was observed in thyroid tissue of patients with PTC, as compared to CD3(-)CD16(+)CD56(dim) NK cells (p=0.046), while CD3(-)CD16(+)CD56(dim) NK cells demonstrated a higher infiltration of NG tissues. CD3(-)CD16(+)CD56(dim) NK cell tissue infiltration positively correlated with advanced stages of PTC. In contrast, the CD3(-)CD16(-)CD56(bright) NK cell population was negatively associated with tumor stage in patients with PTC.CD3(-)CD16(-)CD56(bright) NK cell infiltration seems to be associated with PTC progression. These findings contribute to a better understanding of the immune response in PTC and may lead to novel immunotherapeutic approaches in these patients.

Background: The aim of this study was to test an easy-to-perform flow cytometric (FCM) assay for the routine investigation for diagnosis of paroxysmal nocturnal hemoglobinuria (PNH), through the simultaneous detection of PNH clones on immature reticulocytes (i-RET) and granulocytes. Methods: During the last 5 years, eight patients were diagnosed with PNH in our laboratory, among 90 patients prospectively studied for PNH. The determination of glycosylphosphatidylinositol (GPI) deficient cells on the erythroid lineage was made with a two-color FCM assay of CD71 and CD59, evaluating the PNH clone on i-RET. Three color combinations based on CD66b/CD16/CD45 and CD59/CD24/CD45 were used for the determination of GPI-deficient granulocytes. Results: In all the patients with PNH, the PNH clones determined with CD71(+)CD59(−) red blood cells (RBC) were nearly identical to the respective clones determined with CD16(dim/−)/CD66b(−) and CD59(−)/CD24(−) granulocytes, in contrast to the clones determined with CD59-deficient erythrocytes only, which were significantly lower. Conclusions: Our results indicate that the simultaneous assessment of the PNH clone on CD71(+)/CD59(−)i-RET and CD16(dim/−)/CD66b(−) granulocytes, could offer a reliable method of two series PNH screening, at low cost and with ease of application. © 2012 International Clinical Cytometry Society

Current clinical protocols used for isolation and purification of mesenchymal stem cells (MSC) are based on long-term cultures starting with bone marrow (BM) mononuclear cells. Using a commercially available immunoselection kit for enrichment of MSC, we investigated whether culture of enriched BM-CD105(+) cells could provide an adequate number of pure MSC in a short time for clinical use in the context of graft versus host disease and graft failure/rejection. We isolated a mean of 5.4 × 10(5) ± 0.9 × 10(5) CD105(+) cells from 10 small volume (10-25 ml) BM samples achieving an enrichment >100-fold in MSC. Seeding 2 × 10(3) immunoselected cells/cm(2) we were able to produce 2.5 × 10(8) ± 0.7 × 10(8) MSC from cultures with autologous serum enriched medium within 3 weeks. Neither haematopoietic nor endothelial cells were detectable even in the primary culture cell product. Expanded cells fulfilled both phenotypic and functional current criteria for MSC; they were CD29(+), CD90(+), CD73(+), CD105(+), CD45(-); they suppressed allogeneic T-cell reaction in mixed lymphocyte cultures and retained in vitro differentiation potential. Moreover, comparative genomic hybridization analysis revealed chromosomal stability of the cultured MSC. Our data indicate that adequate numbers of pure MSC suitable for clinical applications can be generated within a short time using enriched BM-CD105(+) cells.

The mechanical integrity of composite materials depends primarily on the interface strength and the defect density of the reinforcement which is the provider of enhanced strength and stiffness. In the case of graphene/ polymer nanocomposites which are characterized by an extremely large interface region, any defects in the inclusion (such as folds, cracks, holes etc.) will have a detrimental effect to the internal strain distribution and the resulting mechanical performance. This conventional wisdom, however, can be challenged if the defect size is reduced beyond the critical size for crack formation to the level of atomic vacancies. In that case, there should be no practical effect on crack propagation and depending on the nature of the vacancies the interface strength may be in fact increase. In this work we employed argon ion (Ar+) bombardment and subsequent exposure to hydrogen (H2) to induce (as revealed by X-ray & Ultraviolet photoelectron spectroscopy (XPS/UPS) and Raman spectroscopy) passivated atomic single vacancies to CVD graphene. The modified graphene was subsequently transferred to PMMA bars and the morphology, wettability and the interface adhesion of the CVD graphene/PMMA system were investigated with Atomic Force Microscopy technique and Raman analysis. The results obtained showed clearly an overall improved mechanical behavior of graphene/polymer interface, since an increase as well a more uniform shift distribution with strain is observed. This paves the way for interface engineering in graphene/polymer systems which, in pristine condition, suffer from premature graphene slippage and subsequent failure.

The authors report a case of intraspinal mass associated with recurrence of B-precursor acute lymphoblastic leukemia in an adolescent male who presented with numb chin syndrome at initial diagnosis of the leukemia. The patient developed sensory changes, later on motor weakness, and eventually paraplegia. An emergent MRI scan showed an intraspinal mass at the level of T9 vertebra. Biopsy obtained during laminectomy revealed a mass composed of lymphoblasts immunophenotypically identical to the patient's known leukemia. Surgical decompression and dexamethasone were ineffective in restoration of the neurological deficits. Intraspinal extramedullary relapses should be considered in the differential diagnosis of leukemic patients with neurological symptoms.

Summary. Shape changes of abnormally deformed red cells in aperture impedance haematology analysers are known to affect MCV, MCHC and haematocrit estimation. However, different counters vary in the manifestation of this effect. We performed a comparative study among five analysers. Three of them are based on impedance without hydrodynamic focusing (Coulter STKR, Cell-Dyn3000 Abbott and K-1000 Sysmex). The other two use hydrodynamic focusing, either with impedance (NE-8000 Sysmex) or two angle laser scatter (H*1 Bayer). A novel method of analysis was applied. Two hundred and three specimens with abnormal red cells and 50 normal specimens (according to ICSH criteria) were assayed. In all samples the PCV was estimated by the reference method without correction for trapped plasma. A true MCHC value was estimated from the mean haemoglobin value and the PCV. The shape effect was assessed by three linear regressions: 1) haematocrit deviations from PCV (corrected for any calibration bias) versus true MCHC; 2) analyser MCHC vs. true MCHC; 3) MCV vs. MCH. The regressions for the analysers with hydrodynamic focusing indicated no significant shape effect. Aperture impedance analysers without focusing varied in their behaviour. The Coulter STKR and the Cell-Dyn3000 both showed strong correlation of haematocrit deviations with true MCHC, poor MCHC correlations and linear MCV-MCH regressions. The K-1000 showed minor indications of such an effect. We conclude that comparative studies are needed to quantitate red cell shape effect errors among various impedance analysers.

Lymphocyte subpopulation imbalances, bone marrow hematopoiesis and histopathology in rituximab-treated lymphoma patients with late-onset neutropenia

A reciprocal t(X;12)(p11;p13) was found as the sole clonal abnormality in biphenotypic leukemia with myeloid and B-lymphoid differentiation. With fluorescence in situ hybridization analysis, the ETV6 gene (previously TEL) was found to be translocated intact to the derivative X chromosome; no MLL and BCR/ABL rearrangements were found. The patient achieved complete remission after induction chemotherapy. To our knowledge, this cytogenetic aberration has not been reported previously as a sole abnormality in hematological malignancies. Its presence may suggest an important role in the pathogenesis of biphenotypic leukemia.

Central issues in intracoronary infusion (ICI) of bone marrow (BM)-cells to damaged myocardium for improving cardiac function are the cell number that is feasible and safe to be administrated as well as the retention of cells in the target area. Our study addressed these issues in eight patients with chronic ischemic cardiomyopathy undergoing ICI of selected BM-progenitors. We could immunomagnetically isolate 0.8 +/- 0.32 x 10(7) CD133(+) cells and 0.75 +/- 0.24 x 10(7) CD133(-)CD34(+) cells from 310 +/- 40 ml BM. After labeling these cells with (99m)Tc-hexamethylpropylenamineoxime, they were infused into the infarct-related artery without any complication. Scintigraphic images 1 (eight patients) and 24 hours (four patients) after ICI revealed an uptake of 9.2% +/- 3.6 and 6.8% +/- 2.4 of the total infused radioactivity in the infarcted area of the heart, respectively; the remaining activity was distributed mainly to liver and spleen. We conclude that through ICI of CD133(+) and CD133(-)CD34(+) BM-progenitors a significant number of them are preferentially attracted to and retained in the chronic ischemic myocardium.

Transferrin receptor (TfR)-1 and 2 mRNA and CD71 (TfR1) expression was analyzed in 118 CLL patients. Ninety-five out of 109 analyzed cases expressed CD71, mostly at a high level; 60% of CD71 (+) cases were IGH-mutated. All samples were TfR1 mRNA (+); TfR2-alpha/beta mRNA was detected in, respectively, 52/102 and 100/109 cases. Competitive RT-PCR showed widely divergent levels of TfR1 mRNA in cases with high CD71 expression, alluding to postrtanscriptional control of TfR1 expression in CLL. The almost uniformly high CD71 expression in CLL is in keeping with the activated status of neoplastic cells, regardless of IGH mutational load.

We present the unusual case of a 16-year-old girl with T-cell acute lymphoblastic leukemia (ALL) with an early thymocyte immunophenotype without myeloid markers, who after 13 months of complete hematological remission relapsed as acute myelogenous leukemia (AML) with minimal differentiation and died of her disease. Whether the AML represented a relapse with lineage switch of the original immature T-cell clone or a new secondary malignancy, could not be proven due to the absence of molecular or clonal markers. This report suggests that a subset of CD7+ T-cell leukemias without mature T-cell antigens (CD4-, CD8-) are minimally differentiated and can relapse as AML.

Immunoglobulin kappa (IGK) locus rearrangements were analyzed in parallel on cDNA/genomic DNA in 188 kappa- and 103 lambda-chronic lymphocytic leukemia (CLL) cases. IGKV-KDE and IGKJ-C-intron-KDE rearrangements were also analyzed on genomic DNA. In kappa-CLL, only 3 of 188 cases carried double in-frame IGKV-J transcripts: in such cases, the possibility that leukemic cells expressed more than one kappa chain cannot be excluded. Twenty-eight kappa-CLL cases also carried nonexpressed (nontranscribed and/or out-of-frame) IGKV-J rearrangements. Taking IGKV-J, IGKV-KDE, and IGKJ-C-intron-KDE rearrangements together, 38% of kappa-CLL cases carried biallelic IGK locus rearrangements. In lambda-CLL, 69 IGKV-J rearrangements were detected in 64 of 103 cases (62%); 24 rearrangements (38.2%) were in-frame. Four cases carried in-frame IGKV-J transcripts but retained monotypic light-chain expression, suggesting posttranscriptional regulation of allelic exclusion. In all, taking IGKV-J, IGKV-KDE, and IGKJ-C-intron-KDE rearrangements together, 97% of lambda-CLL cases had at least 1 rearranged IGK allele, in keeping with normal cells. IG repertoire comparisons in kappa- versus lambda-CLL revealed that CLL precursor cells tried many rearrangements on the same IGK allele before they became lambda producers. Thirteen of 28 and 26 of 69 non-expressed sequences in, respectively, kappa- or lambda-CLL had100% homology to germline. This finding might be considered as evidence for secondary rearrangements occurring after the onset of somatic hypermutation, at least in some cases. The inactivation of potentially functional IGKV-J joints by secondary rearrangements indicates active receptor editing in CLL and provides further evidence for the role of antigen in CLL immunopathogenesis.

Dysfunction of cell membrane is a recognized consequence of the pathogenetic process underlying the β-thalassemia syndromes and it is reasonable to hypothesize that surface structures crucial for the development of erythroid lineage may also be affected. The study included six adult splenectomized patients with β-thalassemia intermedia. Expression of α4β1 integrin (CD49d/CD29), α5β1 integrin (CD49e/CD29) and transferrin receptor (CD71) on peripheral blood and bone marrow erythroblasts and on erythroid precursors grown in vitro was studied by flow cytometry and immunocytochemistry. Serum soluble transferrin receptor levels (sCD71) were also measured with enzyme-linked immunosorbent assay. In β-thalassemic patients, significant reduction of CD49d, CD29 and CD71 expression was found in peripheral blood nucleated red cells, compared to patients presenting with erythroblasts in the circulation because of other diseases. Marrow erythroblasts were also deficient for the same molecules against the erythroblasts in iron deficiency anemia. All molecules tested were greatly diminished on erythroid precursors developed in vitro from the patients' cells. Serum sCD71 levels were much higher in thalassemic patients in comparison to both patients with iron deficiency anemia and healthy individuals. The loss of certain integrins and CD71 from erythroid precursors in β-thalassemia intermedia could be attributed to a generalized membrane dysfunction, perhaps affecting the integrity of their transmembrane domains. The elevation of serum sCD71 levels may be the result of the increased red cell lineage turnover or, alternatively, may indicate increased shedding from the cells to prevent iron overload. In any case, further molecular study of the membrane components is warranted to provide a better understanding of the pathogenetic process in β-thalassemia syndromes.

Immunologic effects of high-dose interferon are still unclear. We have evaluated changes in blood lymphocyte subpopulations, immunoglobulins, and multiple interleukin in patients with high-risk cutaneous melanoma on adjuvant treatment with high-dose interferon and compared pretreatment values with normal controls. Samples were obtained before treatment, 1 month after induction treatment and at 3, 6, and 12 months of maintenance treatment from 24 patients with high-risk melanoma. Lymphocyte subpopulations were measured by flow cytometry and interleukin and immunoglobulin levels by radioimmunoassay. A statistically significant reduction in B-lymphocytes (p < 0.001), natural killer (NK) cells (p = 0.0004), and monocytes (p = 0.04), and an elevation in CD4/ CD8 ratio (p < 0.0001) was observed after I month of intravenous interferon. No changes were seen in CD3, CD4, and CD8 lymphocytes. No changes in interleukin (IL)-2, -4, or -5 were observed during 1 year of treatment. IL-2 pretreatment levels were significantly lower than healthy blood donors (p = 0.001), and IL-5 pretreatment levels were significantly higher (p = 0.0056). IL-10 levels significantly dropped after 6 months of treatment (p = 0.01). Immunoglobulins (IgG, IgA, IgM) remained within normal ranges. Three patients had elevated pretreatment levels of IgE. There is a time- and dose-dependent impact of interferon on numbers of circulating B lymphocytes, NK cells, monocytes, and CD4/CD8 ratio. Defects in cellular and humoral immunity are suggested by the low IL-2 and high IL-5 levels, measured in patients with melanoma as compared with healthy controls.

We report two cases with B cell malignancies (case #1: refractory mantle cell lymphoma; case #2: lymphocyte predominant Hodgkin's disease (LPHD)) who developed neutropenia post-Rituximab therapy in a setting of significant infiltration of the peripheral blood (PB) and bone marrow (BM) by T cells with an immunophenotype of large granular lymphocytes. Possible pathogenetic mechanisms are discussed.

Recurrent chromosomal alterations have been repeatedly reported in cultured human embryonic stem cells (hESCs). The effects of these alterations on the capability of pluripotent cells to differentiate and on growth potential of their specific differentiated derivatives remain unclear. Here, we report that the hESC lines HUES-7 and -9 carrying multiple chromosomal alterations produce in vitro mesenchymal stem cells (MSCs) that show progressive growth arrest and enter senescence after 15 and 16 passages, respectively. There was no difference in their proliferative potential when compared with bone marrow-derived MSCs. Array comparative genomic hybridization analysis (aCGH) of hESCs and their mesenchymal derivatives revealed no significant differences in chromosomal alterations, suggesting that genetically altered hESCs are not selected out during differentiation. Our findings indicate that genetically unstable hESCs maintain their capacity to differentiate in vitro into MSCs, which exhibit an in vitro growth pattern of normal MSCs and not that of transformed cells.

Transferrin receptor (TfR, CD71) is an integral membrane glycoprotein that mediates cellular uptake of iron. In most tissues, TfR expression is correlated positively with proliferation and regulated at the post-transcriptional level. The available data regarding the pattern of TfR gene expression in haematological malignancies are very limited. In the present study, we evaluated TfR gene expression at the molecular level in bone marrow (BM) samples of 44 patients with de novo acute myeloid leukaemia (AML) at diagnosis with BM blasts > 85%. TfR mRNA levels were determined by densitometric analysis of quantitative reverse transcription polymerase chain reaction products corresponding to TfR exons 15–17. Each sample was tested in at least two independent experiments. In 13/44 patients, TfR messages were not detected (this is probably an underestimate as some positive results may be attributed to residual normal erythroid cells present in the samples). In 17/44, TfR mRNA levels were low–intermediate, and were high in the remaining patients (14/44). TfR mRNA positivity was significantly associated with older age. No statistically significant correlations were found either with specific French–American–British (FAB) subtypes or attainment of complete remission, incidence of relapse and survival (after adjusting accordingly for age and FAB subtype). The absence of TfR mRNA transcripts in a significant minority of cases suggests that alternative mechanisms of iron uptake may function in AML blast cells.

PURPOSE: To describe a case of herpes simplex virus type 2 (HSV-2) acute retinal necrosis syndrome (ARN) in a 13-year-old immunocompetent girl. METHODS: Polymerase chain reaction (PCR), cultures, flow cytometry, and cytology were performed on the vitreous sample. RESULTS: Both PCR studies and vitreous cultures revealed HSV-2 as the cause of ARN. Flow cytometry showed CD4+, CD8+, and natural killer cells. The visual outcome of the patient was 20/200. CONCLUSION: Successful culture of HSV-2 from the vitreous specimen in a patient with ARN proved HSV-2 to be one of the causes of ARN. The successful culture of HSV-2 has not been previously reported.