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BACKGROUND: Myelodysplastic syndromes (MDS) represent the most common type of acquired bone marrow failure in adults and is characterized by ineffective maturation of myeloid precursor cells and peripheral cytopenias associated with higher rates of infection, bleeding and transfusion dependence. In higher-risk patients with MDS who relapse or do not respond after standard hypomethylating agent (HMA) therapy, the 2-year survival rate is 15%. METHODS: Here the authors report the feasibility and safety of a novel experimental T-cell therapy called personalized adoptive cell therapy, which selects, immunizes and expands T cells against MDS-specific mutations and is targeted to patient-specific tumor cell neoantigens. Somatic mutations serve as the pathogenic drivers of cancer, including MDS, as these transformative genetic mutations may generate novel immunogenic proteins (i.e., neopeptides and possible neoantigens) that may be targeted therapeutically. RESULTS: The authors demonstrate that the adaptive immune system can be trained ex vivo to recognize neopeptides as neoantigens and that the infusion of culture-expanded, neoantigen-immunized autologous T cells has been feasible and safe in the three patients treated to date. DISCUSSION: The authors report on early results from their first-in-human phase 1 clinical trial that aims to assess the safety and tolerability of this novel form of adoptive T-cell immunotherapy for HMA-refractory patients with higher-risk MDS.
Assuntos
Síndromes Mielodisplásicas , Recidiva Local de Neoplasia , Idoso , Terapia Baseada em Transplante de Células e Tecidos , Humanos , Imunoterapia Adotiva , Síndromes Mielodisplásicas/terapia , Linfócitos TRESUMO
Therapies that utilize immune checkpoint inhibition work by leveraging mutation-derived neoantigens and have shown greater clinical efficacy in tumors with higher mutational burden. Whether tumors with a low mutational burden are susceptible to neoantigen-targeted therapy has not been fully addressed. To examine the feasibility of neoantigen-specific adoptive T-cell therapy, the authors studied the T-cell response against somatic variants in five patients with myelodysplastic syndrome (MDS), a malignancy with a very low tumor mutational burden. DNA and RNA from tumor (CD34+) and normal (CD3+) cells isolated from the patients' blood were sequenced to predict patient-specific MDS neopeptides. Neopeptides representing the somatic variants were used to induce and expand autologous T cells ex vivo, and these were systematically tested in killing assays to determine the proportion of neopeptides yielding neoantigen-specific T cells. The authors identified a total of 32 somatic variants (four to eight per patient) and found that 21 (66%) induced a peptide-specific T-cell response and 19 (59%) induced a T-cell response capable of killing autologous tumor cells. Of the 32 somatic variants, 11 (34%) induced a CD4+ response and 11 (34%) induced a CD8+ response that killed the tumor. These results indicate that in vitro induction of neoantigen-specific T cells is feasible for tumors with very low mutational burden and that this approach warrants investigation as a therapeutic option for such patients.
Assuntos
Síndromes Mielodisplásicas , Neoplasias , Antígenos de Neoplasias/genética , Humanos , Mutação , Síndromes Mielodisplásicas/genética , Síndromes Mielodisplásicas/terapia , Linfócitos TRESUMO
The thermal properties of coarse-grained knotted copolymer rings fluctuating in a highly screening solution are investigated on a simple cubic lattice using the Wang-Landau Monte Carlo algorithm. The rings contain two kinds of monomers A and B with opposite charges that are subjected to short-range interactions. In view of possible applications in medicine and the construction of intelligent materials, it is shown that the behavior of copolymer rings can be tuned by changing both their monomer configuration and topology. We find several phase transitions depending on the monomer distribution. They include the expansion and collapse of the knotted polymer as well as rearrangements leading to metastable states. The temperatures at which these phase transitions are occurring and other features can be tuned by changing the topology of the system. The processes underlying the observed transitions are identified. In knots formed by diblock copolymers, two different classes of behaviors are detected depending on whether there is an excess of monomers of one kind or not. Moreover, we find that the most stable compact states are formed by copolymers in which units of two A monomers are alternated by units of two B monomers. Remarkably, these compact states are in a lamellar phase. The transition from the lamellar to the expanded state produces in the specific heat capacity a narrow and high peak that is centered at temperatures that are much higher than those of the peaks observed in all other monomer distributions.
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BACKGROUND: The incidence of infections associated with cardiac implantable electronic devices (CIEDs) and patient outcomes are not fully known. AIM: To provide a contemporary assessment of the risk of CIEDs infection and associated clinical outcomes. METHODS: In Italy, 18 centres enrolled all consecutive patients undergoing a CIED procedure and entered a 12-months follow-up. CIED infections, as well as a composite clinical event of infection or all-cause death were recorded. RESULTS: A total of 2675 patients (64.3% male, age 78 (70-84)) were enrolled. During follow up 28 (1.1%) CIED infections and 132 (5%) deaths, with 152 (5.7%) composite clinical events were observed. At a multivariate analysis, the type of procedure (revision/upgrading/reimplantation) (OR: 4.08, 95% CI: 1.38-12.08) and diabetes (OR: 2.22, 95% CI: 1.02-4.84) were found as main clinical factors associated to CIED infection. Both the PADIT score and the RI-AIAC Infection score were significantly associated with CIED infections, with the RI-AIAC infection score showing the strongest association (OR: 2.38, 95% CI: 1.60-3.55 for each point), with a c-index = 0.64 (0.52-0.75), p = 0.015. Regarding the occurrence of composite clinical events, the Kolek score, the Shariff score and the RI-AIAC Event score all predicted the outcome, with an AUC for the RI-AIAC Event score equal to 0.67 (0.63-0.71) p < 0.001. CONCLUSIONS: In this Italian nationwide cohort of patients, while the incidence of CIED infections was substantially low, the rate of the composite clinical outcome of infection or all-cause death was quite high and associated with several clinical factors depicting a more impaired clinical status.
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The present paper reports on the production of anthocyanins and xanthones in different in vitro systems of Hypericum perforatum var. angustifolium (sin. Fröhlich) Borkh. Undifferentiated calli and regenerated shoots at different developmental stages were analyzed by applying an extractive and an analytical procedure capable of detecting and quantifying anthocyanins. The findings revealed, for the first time, the co-presence of hypericins and anthocyanins in shoots at initial and more developed stages of H. perforatum var. angustifolium L. Moreover, a high production of xanthones was found in the undifferentiated calli.
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Antocianinas/biossíntese , Hypericum/metabolismo , Brotos de Planta/metabolismo , Xantonas/metabolismo , Antocianinas/análise , Hypericum/citologia , Brotos de Planta/citologia , Regeneração/fisiologia , Xantonas/análiseRESUMO
In this work the dynamics of a chain consisting of a set of beads attached to the ends of segments of fixed lengths is investigated. The chain fluctuates at constant temperature in a viscous medium. For simplicity, all interactions among the beads have been switched off and the number of spatial dimensions has been limited to two. In the limit in which the chain becomes a continuous system, its behavior may be described by a path integral, in which the rigid constraints coming from the infinitesimally small segments are imposed by means of a functional delta function. In this way a model of the dynamics of the chain is obtained, which closely resembles a two-dimensional nonlinear sigma model. The partition function of this generalized nonlinear sigma model is computed explicitly for a ring-shaped chain in the semiclassical approximation. The behavior of the chain at both long and short scales of time and distances is investigated. The connection between the generalized nonlinear sigma model presented here and the Rouse model is discussed.
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The participation of reactive oxygen species (ROS) generated by NOX1 and NOX2/NADPH oxidase has been documented during inflammatory pain. However, the molecular mechanism involved in their activation is not fully understood. We reported earlier a key role of Cyclin-dependent kinase 5 (Cdk5) during inflammatory pain. In particular, we demonstrated that TNF-α increased p35 expression, a Cdk5 activator, causing Cdk5-mediated TRPV1 phosphorylation followed by an increment in Ca2+ influx in nociceptive neurons and increased pain sensation. Here we evaluated if Cdk5 activation mediated by p35 transfection in HEK293 cells or by TNF-α treatment in primary culture of nociceptive neurons could increase ROS production. By immunofluorescence we detected the expression of catalytic subunit (Nox1 and Nox2) and their cytosolic regulators (NOXO1 and p47phox) of NOX1 and NOX2/NADPH oxidase complexes, and their co-localization with Cdk5/p35 in HEK293 cells and in nociceptive neurons. By using a hydrogen peroxide sensor, we detected a significant increase of ROS production in p35 transfected HEK293 cells as compared with control cells. This effect was significantly blocked by VAS2870 (NADPH oxidase inhibitor) or by roscovitine (Cdk5 activity inhibitor). Also by using another ROS probe named DCFH-DA, we found a significant increase of ROS production in nociceptive neurons treated with TNF-α and this effect was also blocked by VAS2870 or by roscovitine treatment. Interestingly, TNF-α increased immunodetection of p35 protein and NOX1 and NOX2/NADPH oxidase complexes in primary culture of trigeminal ganglia neurons. Finally, the cytosolic regulator NOXO1 was significantly translocated to plasma membrane after TNF-α treatment and roscovitine blocked this effect. Altogether these results suggest that Cdk5 activation is implicated in the ROS production by NOX1 and NOX2/NADPH oxidase complexes during inflammatory pain.
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Mycobacterium tuberculosis (Mtb) protein tyrosine phosphatases A and B (PtpA and PtpB) have been recognized as potential molecular targets for the development of new therapeutic strategies against tuberculosis (TB). In this context, we have recently reported that the naturally occurring Diels-Alder-type adduct Kuwanol E is an inhibitor of PtpB (Ki = 1.6 ± 0.1 µM). Here, we describe additional Diels-Alder-type adducts isolated from Morus nigra roots bark that inhibit PtpB at sub-micromolar concentrations. The two most potent compounds, namely Kuwanon G and Kuwanon H, showed Ki values of 0.39 ± 0.27 and 0.20 ± 0.01 µM, respectively, and interacted with the active site of the enzyme as suggested by kinetics and mass spectrometry studies. Molecular docking coupled with intrinsic fluorescence analysis and isothermal titration calorimetry (ITC) further characterized the interaction of these promising PtpB inhibitors. Notably, in an Mtb survival assay inside macrophages, Kuwanon G showed inhibition of Mtb growth by 61.3%. All these results point to the common Diels-Alder-type adduct scaffold, and highlight its relevance for the development of PtpB inhibitors as candidate therapeutics for TB.
Assuntos
Inibidores Enzimáticos/farmacologia , Flavonoides/farmacologia , Morus/química , Mycobacterium tuberculosis/enzimologia , Proteína Tirosina Fosfatase não Receptora Tipo 1/antagonistas & inibidores , Linhagem Celular , Reação de Cicloadição , Relação Dose-Resposta a Droga , Inibidores Enzimáticos/química , Inibidores Enzimáticos/isolamento & purificação , Flavonoides/química , Flavonoides/isolamento & purificação , Humanos , Cinética , Modelos Moleculares , Estrutura Molecular , Proteína Tirosina Fosfatase não Receptora Tipo 1/metabolismo , Relação Estrutura-AtividadeRESUMO
In this paper we study from a nonperturbative point of view the entanglement of two directed polymers subjected to repulsive interactions given by a Dirac delta-function potential. An exact formula of the so-called second moment of the winding angle is derived. This result is used to provide a thorough analysis of entanglement phenomena in the classical system of two polymers subjected to repulsive interactions and related problems. No approximation is made in treating the constraint on the winding angle and the repulsive forces. In particular, we investigate how repulsive forces influence the entanglement degree of the two-polymer system. In the limit of ideal polymers, in which the interactions are switched off, we show that our results are in agreement with those of previous works.
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Two new xanthone derivatives, 1-hydroxy-5,6,7-trimethoxyxanthone and 3-O-methylpaxanthone were isolated from callus of Hypericum perforatum subsp. perforatum together with the known paxanthone, cadensin G, 1-hydroxy-6,7-dimethoxyxanthone, 1,3,6,7-tetrahydroxyxanthone, and 1,3,5,6-tetrahydroxyxanthone. The structures of the new compounds were established by spectroscopic methods.
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Hypericum/química , Xantonas/isolamento & purificação , Hypericum/crescimento & desenvolvimento , Espectroscopia de Ressonância Magnética , Xantonas/químicaRESUMO
A new isoflavone 5,7,4'-trihydroxy-3'-(3-hydroxy-3-methylbutyl)isoflavone (isowigtheone hydrate) (1), together with six known isoflavones 2-7 and (-)epicatechin, were isolated from the root barks of Brosimum utile. Their structures were established on the basis of spectroscopic evidence. The in vitro cytotoxic activity of the new compound 1 was evaluated against cell lines MCF7 (human breast carcinoma), PC3 (human prostate carcinoma), HT29 (human colon cancer) and human dermis fibroblasts.
Assuntos
Antineoplásicos Fitogênicos/química , Isoflavonas/química , Moraceae/química , Casca de Planta/química , Raízes de Plantas/química , Antineoplásicos Fitogênicos/isolamento & purificação , Antineoplásicos Fitogênicos/farmacologia , Linhagem Celular Tumoral , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Isoflavonas/isolamento & purificação , Isoflavonas/farmacologia , Estrutura MolecularRESUMO
BACKGROUND: Experts and scientific society guidelines recommend that rapid on-site evaluation (ROSE) be used with endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) to optimize lung cancer genotyping, but no comparative trial has been carried out to confirm and quantify its usefulness. METHODS: To assess the influence of ROSE on the yield of EBUS-TBNA for a multigene molecular analysis of lung cancer samples, consecutive patients with suspected or known advanced lung cancer were randomized to undergo EBUS-TBNA without ROSE (EBUS arm) or with ROSE (ROSE arm). The primary end point was the rate of the successful accomplishment of the institution's clinical protocol for molecular profiling of nonsquamous non-small cell lung cancer (EGFR and KRAS testing, followed by ALK testing for tumors with EGFR and KRAS wild-type status). RESULTS: Complete genotyping was achieved in 108 of 126 patients (85.7%) (90.8% in the ROSE arm vs 80.3% in the EBUS arm, P = .09). The patients in the ROSE arm were less likely to have samples that could be used only for pathologic diagnosis because of minimal tumor burden (0 vs 6, P = .05), and were more likely to have the bronchoscopy terminated after a single biopsy site (58.9% vs 44.1%, P = .01). CONCLUSIONS: ROSE prevents the need for a repeat invasive diagnostic procedure aimed at molecular profiling in at least one out of 10 patients with advanced lung cancer and significantly reduces the risk of retrieving samples that can be used only for pathologic subtyping because of minimal tumor burden. TRIAL REGISTRY: ClinicalTrials.gov; No.: NCT01799382; URL: www.clinicaltrials.gov.
Assuntos
Broncoscopia/métodos , Carcinoma Pulmonar de Células não Pequenas , Aspiração por Agulha Fina Guiada por Ultrassom Endoscópico/métodos , Neoplasias Pulmonares , Idoso , Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/patologia , Receptores ErbB/genética , Feminino , Genótipo , Técnicas de Genotipagem , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Masculino , Estadiamento de Neoplasias , Valor Preditivo dos Testes , Proteínas Proto-Oncogênicas p21(ras)/genética , Sensibilidade e Especificidade , Manejo de Espécimes/métodos , Carga TumoralRESUMO
Protein tyrosine phosphatase B (PtpB) is one of the virulence factors secreted into the host cell by Mycobacterium tuberculosis. PtpB attenuates host immune defenses by interfering with signal transduction pathways in macrophages and, therefore, it is considered a promising target for the development of novel anti-tuberculosis drugs. Here we report the discovery of natural compound inhibitors of PtpB among an in house library of more than 800 natural substances by means of a multidisciplinary approach, mixing in silico screening with enzymatic and kinetics studies and MS assays. Six natural compounds proved to inhibit PtpB at low micromolar concentrations (< 30 µM) with Kuwanol E being the most potent with K i = 1.6 ± 0.1 µM. To the best of our knowledge, Kuwanol E is the most potent natural compound PtpB inhibitor reported so far, as well as it is the first non-peptidic PtpB inhibitor discovered from natural sources. Compounds herein identified may inspire the design of novel specific PtpB inhibitors.
Assuntos
Produtos Biológicos/farmacologia , Descoberta de Drogas , Inibidores Enzimáticos/farmacologia , Mycobacterium tuberculosis/enzimologia , Proteínas Tirosina Fosfatases/antagonistas & inibidores , Sequência de Aminoácidos , Proteínas de Bactérias/antagonistas & inibidores , Proteínas de Bactérias/metabolismo , Produtos Biológicos/química , Inibidores Enzimáticos/química , Humanos , Concentração Inibidora 50 , Cinética , Modelos Moleculares , Dados de Sequência Molecular , Mycobacterium tuberculosis/efeitos dos fármacos , Mapeamento de Peptídeos , Ligação Proteica/efeitos dos fármacos , Proteína Tirosina Fosfatase não Receptora Tipo 1/antagonistas & inibidores , Proteína Tirosina Fosfatase não Receptora Tipo 1/metabolismo , Proteínas Tirosina Fosfatases/química , ProteóliseRESUMO
A coronary angiography performed for the occurrence of atypical chest pain allowed us to discover a coronary-right ventricular fistula as a rare complication of myocardial penetration by a tined ventricular catheter implanted some years earlier.
Assuntos
Estimulação Cardíaca Artificial/efeitos adversos , Vasos Coronários , Ventrículos do Coração , Marca-Passo Artificial/efeitos adversos , Fístula Vascular/etiologia , Antagonistas Adrenérgicos beta/uso terapêutico , Idoso , Fibrilação Atrial/complicações , Fibrilação Atrial/terapia , Humanos , Masculino , Resultado do Tratamento , Fístula Vascular/tratamento farmacológicoRESUMO
Cosmic rays represent one of the most fascinating research themes in modern astronomy and physics. Significant progress is being made toward an understanding of the astrophysics of the sources of cosmic rays and the physics of interactions in the ultrahigh-energy range. This is possible because several new experiments in these areas have been initiated. Cosmic rays may hold answers to a great number of fundamental questions, but they also shape our natural habitat and influence the radiation environment of our planet Earth. The importance of the study of cosmic rays has been acknowledged in many fields, including space weather science and astrobiology. Here, we concentrate on the astrobiological aspects of cosmic rays with regard to the enormous amount of new data available, some of which may, in fact, improve our knowledge about the radiation of cosmic origin on Earth. We focus on fluxes arriving at Earth and doses received, and will guide the reader through the wealth of scientific literature on cosmic rays. We have prepared a concise and self-contained source of data and recipes useful for performing interdisciplinary research in cosmic rays and their effects on life on Earth.
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Radiação Cósmica , Planeta Terra , Exobiologia , Genes/efeitos da radiação , Mutação , RadiometriaRESUMO
Correctional facilities host a disproportionately high prevalence of HBV, HCV and HIV infection. We evaluated the prevalence of HBV and/or HCV co-infection among HIV-infected inmates entering our correctional facility. Over a 30-month period, 173 consecutive HIV-infected inmates entered our institution and were evaluated. HCV co-infection was observed in more than 90% of the tested HIV-infected inmates, past HBV infection in 77.4% and active HBV co-infection in 6.7%; triple coinfection (HIV, HCV and HBs-Ag positivity) was seen in 6.1% of them. Given the observed high prevalence of co-infection, testing for HBV and HCV in all HIV-infected inmates at entry in any correctional system is recommended to identify those in need of specific care and/or preventing interventions.
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Infecções por HIV/epidemiologia , Hepatite B/epidemiologia , Hepatite C/epidemiologia , Prisioneiros/estatística & dados numéricos , Distribuição de Qui-Quadrado , Infecções por HIV/diagnóstico , Infecções por HIV/virologia , Hepatite B/diagnóstico , Hepatite B/virologia , Hepatite C/diagnóstico , Hepatite C/virologia , Humanos , Itália/epidemiologia , Masculino , Programas de Rastreamento , Prevalência , Estudos Prospectivos , Estudos Soroepidemiológicos , Abuso de Substâncias por Via Intravenosa/epidemiologia , Abuso de Substâncias por Via Intravenosa/virologiaRESUMO
During the implantation of a definitive pacemaker for complete atrioventricular block via the left subclavian vein, a rare and abnormal origin of the superior vena cava was detected. No such finding has ever been reported, leading us to adopt the contralateral approach.
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Anormalidades Cardiovasculares/complicações , Bloqueio Cardíaco/complicações , Achados Incidentais , Veia Cava Superior/anormalidades , Idoso de 80 Anos ou mais , Procedimentos Cirúrgicos Cardíacos , Anormalidades Cardiovasculares/diagnóstico , Anormalidades Cardiovasculares/fisiopatologia , Circulação Coronária , Diagnóstico Diferencial , Eletrocardiografia , Feminino , Bloqueio Cardíaco/fisiopatologia , Bloqueio Cardíaco/cirurgia , Sistema de Condução Cardíaco/fisiopatologia , Humanos , Período Intraoperatório , Marca-Passo Artificial , Flebografia , Fluxo Sanguíneo Regional , Síndrome da Veia Cava Superior/diagnóstico , Tomografia Computadorizada por Raios X , Veia Cava Superior/diagnóstico por imagem , Veia Cava Superior/fisiopatologiaRESUMO
The unique advantage of genetic engineering techniques for the design and development of polymers for controlled gene delivery lies in exquisite control over polymer structure. In this article we report the biosynthesis and characterization of a series of new silk-elastinlike protein polymers (SELPs), namely, SELP415K, with larger elastin blocks per monomer unit than SELP47K previously studied for matrix-mediated gene delivery. A new cloning strategy was used, where a block of eight elastin units (8E) was integrated into the existing DNA sequence of SELP47K monomer genes using appropriate restriction endonuclease recognition sites. Following random multimerization, multimer gene segments of desired size were selected, expressed, and purified on Ni-agarose columns. The molecular weight and sequence composition of the purified SELPs were determined by MALDI-TOF and amino acid analysis, respectively. The influence of structural changes on the rheological properties of the polymers was investigated. In addition, hydrogel disks were prepared from 47K and 415K-8mer polymer solutions, and the effects of cure time and environmental conditions on the hydrogel equilibrium swelling ratio as a function of polymer composition were studied. DNA sequencing and agarose gel electrophoresis confirmed the successful cloning of the monomer gene segment of SELP415K consisting of 312 bp. Random concatemerization of SELP415K monomer gene segments resulted in a library of SELP415K multimer sequences of 6, 8, and 10 repeats respectively, each yielding a polymer with exact molecular weight and sequence. Rheometric measurements showed that both complex shear modulus (G*) and gelation point were influenced by polymer composition. Equilibrium swelling studies on hydrogel disks prepared from 47K and 415K-8mer polymer solutions showed that changes in polymer composition resulted in different gelation patterns and increased sensitivity toward changes in temperature and ionic strength but not pH. Together these results demonstrate the potential of recombinant techniques in engineering polymers with defined structures which allows the study of the structural parameters affecting matrix-mediated delivery of genes and bioactive agents.
Assuntos
Materiais Biocompatíveis/química , Elastina/química , Técnicas de Transferência de Genes , Engenharia Genética/métodos , Vetores Genéticos , Polímeros/química , Seda/química , Sequência de Aminoácidos , Sequência de Bases , Clonagem Molecular , DNA/química , Elastina/genética , Eletroforese em Gel de Poliacrilamida , Escherichia coli/metabolismo , Hidrogel de Polietilenoglicol-Dimetacrilato/química , Hidrogéis/química , Concentração de Íons de Hidrogênio , Íons , Dados de Sequência Molecular , Níquel/química , Oligonucleotídeos/química , Plasmídeos/química , Proteínas Recombinantes/química , Sefarose/química , Análise de Sequência de DNA , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz , Temperatura , Fatores de TempoRESUMO
Two new Diels-Alder type adducts, named sorocein L (1) and sorocein M (2), together with the known kuwanon J and mulberrofuran O, were isolated from the methanolic extract of the roots of Sorocea ilicifolia. Sorocein L shows the same substitution pattern of sorocein M and corresponds to its ketalized derivative.