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Lentiviral vectors (LV) are efficient vehicles for in vivo gene delivery to the liver. LV integration into the chromatin of target cells ensures their transmission upon proliferation, thus allowing potentially life-long gene therapy following a single administration, even to young individuals. The glycoprotein of the vesicular stomatitis virus (VSV.G) is widely used to pseudotype LV, as it confers broad tropism and high stability. The baculovirus-derived GP64 envelope protein has been proposed as an alternative for in vivo liver-directed gene therapy. Here, we perform a detailed comparison of VSV.G- and GP64-pseudotyped LV in vitro and in vivo. We report that VSV.G-LV transduced hepatocytes better than GP64-LV, however the latter showed improved transduction of liver sinusoidal endothelial cells (LSEC). Combining GP64-pseudotyping with the high surface content of the phagocytosis inhibitor CD47 further enhanced LSEC transduction. Coagulation factor VIII (FVIII), the gene mutated in hemophilia A, is naturally expressed by LSEC, thus we exploited GP64-LV to deliver a FVIII transgene under the control of the endogenous FVIII promoter and achieved therapeutic amounts of FVIII and correction of hemophilia A mice. Synopsis: Lentiviral vectors bearing the baculovirus GP64 surface protein and high content of the phagocytosis inhibitor CD47 efficiently transduce liver endothelial cells in vivo and allow stable long-term correction of mice with hemophilia A, both when treated as adults and as newborns. Development of an in vitro phagocytosis assay, exploiting fluorescent virions and advanced imaging techniques, suitable to assess the susceptibility to phagocytosis of different LV versions in different experimental settings. Efficient tropism switch of LV from hepatocytes to liver endothelial cells in vitro and in vivo and shielding from phagocytosis. Reconstitution of FVIII activity in the normal range by a single intravenous administration of GP64-LV equipped with the endogenous endothelial-specific FVIII promoter. Proof-of-concept of stable long-term gene therapy for hemophilia A in both adult and newborn mice. Lentiviral vectors bearing the baculovirus GP64 surface protein and high content of the phagocytosis inhibitor CD47 efficiently transduce liver endothelial cells in vivo and allow stable long-term correction of mice with hemophilia A, both when treated as adults and as newborns. [ABSTRACT FROM AUTHOR]

Common variable immunodeficiency (CVID) is the most frequent primary antibody deficiency whereby follicular helper T (Tfh) cells fail to establish productive responses with B cells in germinal centers. Here, we analyzed the frequency, phenotype, transcriptome, and function of circulating Tfh (cTfh) cells in CVID patients displaying autoimmunity as an additional phenotype. A group of patients showed a high frequency of cTfh1 cells and a prominent expression of PD‐1 and ICOS as well as a cTfh mRNA signature consistent with highly activated, but exhausted, senescent, and apoptotic cells. Plasmatic CXCL13 levels were elevated in this group and positively correlated with cTfh1 cell frequency and PD‐1 levels. Monoallelic variants in RTEL1, a telomere length‐ and DNA repair‐related gene, were identified in four patients belonging to this group. Their blood lymphocytes showed shortened telomeres, while their cTfh were more prone to apoptosis. These data point toward a novel pathogenetic mechanism in CVID, whereby alterations in DNA repair and telomere elongation might predispose to antibody deficiency. A Th1, highly activated but exhausted and apoptotic cTfh phenotype was associated with this form of CVID. [ABSTRACT FROM AUTHOR]

Introduction: Health professions are heavily engaged facing the current threat of SARS‐CoV‐2 (COVID‐19). Although there are many diagnostic tools, an accurate and rapid laboratory procedure for diagnosing COVID‐19 is recommended. We focused on platelet parameters as the additional biomarkers for clinical diagnosis in patients presenting to the emergency department (ED). Materials and Methods: Five hundred and sixty‐one patients from February to April 2020 have been recruited. Patients were divided into three groups: (N = 50) COVID‐19 positive and (N = 21) COVID‐19 negative with molecular testing, (N = 490) as reference population without molecular testing. A Multiplex rRT‐PCR from samples collected by nasopharyngeal swabs was performed and the hematological data collected. Results: We detected a mild anemia in COVID‐19 group and lymphopenia against reference population: hemoglobin (g/dL) 13.0 (11.5‐14.8) versus 13.9 (12.8‐15.0) (P =.0135); lymphocytes (109/L) 1.24 (0.94‐1.73) versus 1.99 (1.49‐2.64) (P <.0001). In addition, abnormal platelet parameters as follows (COVID group vs reference population): PLT (×109/L) 209 (160‐258) vs 236 (193‐279) (P =.0239). IPF (%) 4.05 (2.5‐5.9) versus 3.4 (2.2‐4.9) (P =.0576); H‐IPF (%) 1.25 (0.8‐2.2) versus 0.95 (0.6‐1.5) (P =.0171) were identified. In particular, COVID positive group had a high H‐IPF/IPF Ratio compared to reference population [0.32 (0.29‐0.36) versus 0.29 (0.26‐0.32), respectively, (P =.0003)]. Finally, a PLT difference of nearly 50 × 109/L between pre/postCOVID‐19 sampling for each patient was found (N = 42) (P =.0194). Conclusions: COVID‐19 group results highlighted higher IPF and H‐IPF values, with increased H‐IPF/IPF Ratio, associated to PLT count reduction. These findings shall be adopted for a timely diagnosis of patients upon hospital admission. [ABSTRACT FROM AUTHOR]

The world of lubricants is driven by the constant pursuit of improved performance in response of the requests of new engine generations. Engine oils play a critical role as lubricants in mitigating wear, reducing friction and ensuring optimal engine operation under diverse conditions. Modern commercial engine oils are complex formulations, comprising of a base oil, generally coming from petroleum sources, formulated with specific, important additives able to optimize the viscosity, thickening and shear stress in the operating temperature range. Such additives are produced in the thousand tons per year scale range. The most important class of additives for modern lubrication is made of organic polymers with variable architectures and topologies, generally referred as “viscosity modifiers” (VMs): they act as “moderators” of viscosity at different working temperatures. The tremendous advances in polymer science have been reflected in the realm of VMs, allowing the commercialization of products obtained by controlled polymerization techniques, and the experimentation of a broad variety of different macromolecular architectures and topologies as VMs. In this review we introduce the reader, together with the basic principles of viscosity modification and thermal‐dependent rheological response, to the fascinating chemistry towards the improvement of VMs, through optimization of macromolecular design and architecture. [ABSTRACT FROM AUTHOR]

Allogeneic transplantation is the only potentially curative strategy for myelofibrosis, even in the era of new drugs that so far only mitigate symptoms. The choice to proceed to allogeneic transplantation is based on several variables including age, disease phase, degree of splenomegaly, donor availability, comorbidities and iron overload. These factors, along with conditioning regimen and time to transplantation, may influence the outcome of ASCT. We report 14 patients affected by myelofibrosis with a median age of 57 years (range, 41-76) receiving a treosulfan-fludarabine based reduced toxicity conditioning. Patients (pts) received a stem cell transplantation from an HLA identical (n = 10) or matched unrelated donor (n = 4). All pts had a complete myeloablation followed by engraftment and in 12 out of 13 evaluated pts donor chimerism was 100% at 1 month. In most cases a reduction of splenomegaly and a reduction (or resolution) of bone marrow fibrosis was observed. After a median follow-up of 39 months (range, 3-106), the 3-year probability of overall survival and disease free survival was 54 +/- 14% and 46 +/- 14%, respectively. The cumulative incidence of non-relapse mortality at 2 years was 39 +/- 15%. Causes of non-relapse mortality were: infection (n = 2), GvHD (n = 2) and haemorrhage (n = 1). We can conclude that a treosulfan and fludarabine based conditioning has a potent myeloablative and anti-disease activity although non-relapse mortality remains high in this challenging clinical setting. Copyright © 2014 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Relapse represents the most significant cause of failure of allogeneic hematopoietic stem cell transplantation ( HSCT) for FLT3- ITD-positive acute myeloid leukemia ( AML), and available therapies are largely unsatisfactory. In this study, we retrospectively collected data on the off-label use of the tyrosine kinase inhibitor sorafenib, either alone or in association with hypomethylating agents and adoptive immunotherapy, in 13 patients with post-transplantation FLT3- ITD-positive AML relapses. Hematological response was documented in 12 of 13 patients (92%), and five of 13 (38%) achieved complete bone marrow remission. Treatment was overall manageable in the outpatient setting, although all patients experienced significant adverse events, especially severe cytopenias (requiring a donor stem cell boost in five patients) and typical hand-foot syndrome. None of the patients developed graft-vs.-host disease following sorafenib alone, whereas this was frequently observed when this was given in association with donor T-cell infusions. Six patients are alive and in remission at the last follow-up, and four could be bridged to a second allogeneic HSCT, configuring a 65 ± 14% overall survival at 100 d from relapse. Taken together, our data suggest that sorafenib might represent a valid treatment option for patients with FLT3- ITD-positive post-transplantation relapses, manageable also in combination with other therapeutic strategies. [ABSTRACT FROM AUTHOR]

BACKGROUND In allogeneic hematopoietic stem cell (HSC) transplantation, the collection of an appropriate number of HSCs while maintaining a high level of safety for healthy donors is fundamental. Inadequate HSC mobilization can be seen with the standard use of granulocyte-colony-stimulating (G-CSF). Plerixafor (PL) is a chemokine receptor CXC Type 4-stromal-derived factor 1 inhibitor; its HSC-mobilizing properties are synergistic with G-CSF in poor mobilizing patients. The use of PL as adjuvant or alternative to G-CSF in healthy donors has shown a good safety profile but is so far off-label. STUDY DESIGN AND METHODS We report 10 healthy HSC donors treated with PL because of insufficient response to G-CSF alone or contraindication to G-CSF. Eight donors did not mobilize enough CD34+ cells with G-CSF alone because poor mobilizers or because insufficient HSCs were harvested according to the clinical need of the patient; in two cases G-CSF administration and marrow harvest were unfeasible or contraindicated in the donor. RESULTS The use of PL for mobilization increased the number of circulating CD34+ cells by 2.8-fold and the CD34+/kg collection by 3.0-fold. Only mild adverse events were reported (bone pain or discomfort) and not univocally attributable to PL. Rate of engraftment and graft-versus-host disease were similar to those seen in recipients of grafts from G-CSF only-mobilized donors. CONCLUSION We exposed 10 allogeneic donors to mobilization with PL. PL was well tolerated in all cases and ensured procurement of an adequate graft for transplantation resulting in a normal hematopoietic engraftment. [ABSTRACT FROM AUTHOR]

Neuromyelitis optica is a rare neurological autoimmune disorder characterized by a poor prognosis. Immunosuppression can halt disease progression, but some patients are refractory to multiple treatments, experiencing frequent relapses with accumulating disability. Here we report on durable clinical remissions after allogeneic hematopoietic stem cell transplantation in 2 patients suffering from severe forms of the disease. Immunological data evidenced disappearance of the pathogenic antibodies and regeneration of a naive immune system of donor origin. These findings correlated with evident clinical and radiological improvement in both patients, warranting extended clinical trials to investigate this promising therapeutic option. ANN NEUROL 2014;75:447-453 [ABSTRACT FROM AUTHOR]

Despite a high proportion of patients with primary CNS lymphoma (PCNSL) experiences failure after/during first-line treatment, a few studies focused on salvage therapy are available, often with disappointing results. Herein, we report feasibility and activity of a combination of rituximab, ifosfamide and etoposide (R-IE regimen) in a multicentre series of patients with PCNSL relapsed or refractory to high-dose methotrexate-based chemotherapy. We considered consecutive HIV-negative patients ≤75 years old with failed PCNSL treated with R-IE regimen (rituximab 375 mg/m2, day 0; ifosfamide 2 g/m2/day, days1-3; etoposide 250 mg/m2, day 1; four courses). Twenty-two patients (median age 60 years; range 39-72; male/female ratio: 1:4) received R-IE as second-line ( n = 18) or third-line ( n = 4) treatment. Eleven patients had refractory PCNSL, and 11 had relapsing disease. Twelve patients had been previously irradiated. Sixty (68%) of the 88 planned courses were actually delivered; only one patient interrupted R-IE because of toxicity. Grade 4 hematological toxicity was manageable; a single case of grade 4 non-hematological toxicity (transient hepatotoxicity) was recorded. Response was complete in six patients and partial in three (overall response rate = 41%; 95%CI: 21-61%). Seven patients were successfully referred to autologous peripheral blood stem cell collection; four responders were consolidated with high-dose chemotherapy supported by autologous stem cell transplant. At a median follow-up of 24 months, eight responders did not experience relapse, two of them died of neurological impairment while in remission. Six patients are alive, with a 2-year survival after relapse of 25 ± 9%. We concluded that R-IE is a feasible and active combination for patients with relapsed/refractory PCNSL. This regimen allows stem cell collection and successful consolidation with high-dose chemotherapy and autologous transplant. Copyright © 2012 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

A new family of ethylene-based copolymers with controlled amounts of a norbornene comonomer (NArOH) bearing a stabilizing antioxidant functionality (2,6-di- tert-butyl phenol) was prepared. Due to unavoidable exo/ endo equilibrium operative in NArOH comonomer, a complete and detailed NMR assignment of the structure of the prepared ethylene/NArOH copolymers was carried out for the determination of the exo/ endo ratio inside the polymer. These novel functionalized comonomers can be considered suitable starting material for preparing ethylene-based copolymers, with tunable comonomer content, as non-releasing macromolecular antioxidant additives for specific application in safe food and/or drug packaging © 2012 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem, 2012 [ABSTRACT FROM AUTHOR]

Central nervous system (CNS) dissemination is a rare (4-5%) but usually fatal complication of aggressive lymphomas. Prophylaxis modalities to prevent CNS dissemination in aggressive lymphomas cannot be widely applied to every lymphoma patient since it is associated with increased risk of neurotoxicity. Therefore, identification of high-risk patients as the best candidates to receive CNS prophylaxis constitutes a major endpoint in the management of these malignancies. Various risk factors and models for CNS recurrence have been described. Parameters reflecting the extent and proliferation of the disease, like elevated serum lactate dehydrogenase levels, involvement of multiple extranodal sites, advanced stage and high age-adjusted International Prognostic Index (IPI) score, as well as the involvement of specific anatomic sites, like testes, orbit, paranasal sinuses, have been identified and confirmed as important to predict CNS dissemination. Management of this complication in aggressive lymphomas with conventional-dose chemotherapy is associated with disappointing results, while some preliminary but encouraging experiences suggest a potential role of high-dose chemotherapy and stem cell transplantation. The analysis of recent clinical studies could lead to advancement in the prognosis of aggressive lymphomas, but several questions regarding the optimum chemotherapy combination, the best conditioning regimen and the role of radiation therapy and intrathecal chemotherapy remain still unanswered. The purposes of the present review are to critically analyse current data on the risk of CNS dissemination in aggressive lymphomas, the clinical presentation of secondary CNS lymphomas and the efficacy of CNS prophylaxis as well as to discuss the available therapeutic options for this devastating event. Copyright © 2009 John Wiley & Sons, Ltd. [ABSTRACT FROM AUTHOR]

Background: Aim of our study is to evaluate the role of T-wave alternans (TWA) to stratify the risk of sudden cardiac death in athletes (Ath) with complex ventricular arrhythmias (VA), and to document a possible correlation between TWA and electrophysiological testing (EPS) results. Methods: We studied 85 Ath with VA (61 M, mean age 32 ± 11 years). In all cases a cardiological evaluation was performed, including TWA and EPS. The patients were evaluated during a follow-up of 30 ± 21 months. The end point was the occurrence of sudden death (SD) or malignant ventricular tachyarrhythmias (VT). Results: TWA was negative in 57 Ath (68%), positive in 15 (18%) and indeterminate in 13 (14%). All subjects with negative TWA did not show induction of VT at EPS, with significant correlation between negative TWA and negative EPS (P < 0.001). All Ath with positive TWA also had VT induced by a EPS, with significant correlation (P < 0.001). By contrast, our data did not show significant correlation between indeterminate TWA and positive or negative EPS. However, there was significant correlation between abnormal TWA test (positive + indeterminate) and inducibility of VT at EPS (P < 0.001). During follow-up we observed a significant difference in end point occurrence (VT or SD) between Ath with negative or abnormal TWA and between Ath with negative or positive EPS. Conclusion: TWA confirm its role as a simple and noninvasive test, and it seems useful for prognostic stratification of Ath with VA. [ABSTRACT FROM AUTHOR]

We investigated the relationship between plasma renin activity (PRA) and serum ([sNa+]) and urinary ([uNa+]) sodium concentrations in 124 congestive heart failure (CHF) patients (II–IV NYHA class) and 20 healthy subjects. According to PRA (> or <3 ng ml−1 h−1) and [sNa+] (> or <135 mEq l−1), patients were classified as Group A (normal PRA and normal [sNa+], n=39), Group B (increased PRA and normal [sNa+], n=62) and Group C (low [sNa+], n=23). Measurements were performed at rest and, in 26 cases, after extracorporeal ultrafiltration (UF). At rest, [sNa+] and [uNa+], and their difference ([sNa+]−[uNa+]), were linearly correlated with PRA, but the values did not allow differentiation of control subjects from patients or differentiation of patients with from those without renin–angiotensin system (RAS) activation. Conversely, the [sNa+]/[uNa+] ratio showed the best correlation with PRA (r=0.79, P<0.0001). UF-induced PRA changes were linearly correlated with [sNa+]/[uNa+] ratio changes (r=0.67, P=0.002), but not with those of [sNa+], [uNa+] and [sNa+]−[uNa+]. In CHF, the [sNa+]/[uNa+] ratio best correlates with PRA and reflects the basal activity as well as the rapid changes (as those induced by UF) of the RAS. Therefore, it can be considered a strong and easily available marker of PRA. [Copyright &y& Elsevier]

BackgroundWe investigated the lung contribution to circulating noradrenaline (NA) homeostasis. Evaluation of the transpulmonary NA gradient, related to the NA amount entering the lungs, is potentially important, mainly regarding clinical conditions, such as congestive heart failure (CHF), that are associated with excessive circulating NA. Materials and methods15 moderate (group 1) and 15 severe (group 2) CHF patients, and 10 normal individuals had determination of NA transpulmonary gradient in the baseline and during rise (exercise, in normals and group 1) or fall (withdrawal from plasma by ultrafiltration, in group 2) of plasma NA. ResultsNA gradient (pg mL-1) at rest was 30 ± 3 in normals, 21 ± 6 in group 1 and 5 ± 8 in group 2. Increase of NA concentration in the mixed venous blood with exercise was paralleled by depression of the transpulmonary gradient. Pulmonary arteriovenous difference disappeared when NA entering the lungs averaged 1300 pg mL-1. In group 2, ultrafiltration lowered NA in the mixed venous blood from 1225 ± 213 to 718 ± 182, which caused transpulmonary gradient to increase from 5 ± 8 to 22 ± 9. ConclusionsTranspulmonary gradient of NA diminishes when NA entering the lungs increases, and 1300 pg mL-1 in the pulmonary artery is, both in patients and normal subjects, the level at which gradient disappears; which likely reflects cessation of NA uptake or achievement of a balance between lung uptake and production. This may have physiological and pathological implications. [ABSTRACT FROM AUTHOR]

It is still a matter of controversy as to whether the patients paced for atrioventricular block (AVB) have different prognosis and survival rates than those paced for Sick Sinus Syndrome (SSS). We have compared the survival rates of 962 AVB patients (group A) with that of 283 SSS patients (group B) who underwent pacemaker implantation during the period January 2968 to December 2986. The survival rate graphs of the examined groups were calculated using the actuarial method and the differences in the survival rates between the groups were evaluated using the Logrank test. Our results show that SSS patients have a higher survival rate than AV block with a difference on the rate of survival between the two groups reaching the borderline of statistical significance. Multivariate discriminant analysis was then used to assess that of the parameters (i.e., age at the time of implantation, sex, electrophysiological indication to pacing, etiology or pacing mode) could have had the main influence upon mortality and the different pattern of the survival rate graph within the two groups of patients. Our data show that survival is mostly related to age, pacing mode and, although more slightly, to underlying heart disease; the electrophysiological indication to pacing, instead, does not significantly influence it. [ABSTRACT FROM AUTHOR]