Inhibitors of class I HDACs and of FLT3 combine synergistically against leukemia cells with mutant FLT3

Acute myeloid leukemia (AML) with mutations in the FMS-like tyrosine kinase (FLT3) is a clinically unresolved problem. AML cells frequently have a dysregulated expression and activity of epigenetic modulators of the histone deacetylase (HDAC) family. Therefore, we tested whether a combined inhibition of mutant FLT3 and class I HDACs is effective against AML cells. Low nanomolar doses of the FLT3 inhibitor (FLT3i) AC220 and an inhibition of class I HDACs with nanomolar concentrations of FK228 or micromolar doses of the HDAC3 specific agent RGFP966 synergistically induce apoptosis of AML cells that carry hyperactive FLT3 with an internal tandem duplication (FLT3-ITD). This does not occur in leukemic cells with wild-type FLT3 and without FLT3, suggesting a preferential toxicity of this combination against cells with mutant FLT3. Moreover, nanomolar doses of the new FLT3i marbotinib combine favorably with FK228 against leukemic cells with FLT3-ITD. The combinatorial treatments potentiated their suppressive effects on the tyrosine phosphorylation and stability of FLT3-ITD and its downstream signaling to the kinases ERK1/ERK2 and the inducible transcription factor STAT5. The beneficial pro-apoptotic effects of FLT3i and HDACi against leukemic cells with mutant FLT3 are associated with dose- and drug-dependent alterations of cell cycle distribution and DNA damage. This is linked to a modulation of the tumor-suppressive transcription factor p53 and its target cyclin-dependent kinase inhibitor p21. While HDACi induce p21, AC220 suppresses the expression of p53 and p21. Furthermore, we show that both FLT3-ITD and class I HDAC activity promote the expression of the checkpoint kinases CHK1 and WEE1, thymidylate synthase, and the DNA repair protein RAD51 in leukemic cells. A genetic depletion of HDAC3 attenuates the expression of such proteins. Thus, class I HDACs and hyperactive FLT3 appear to be valid targets in AML cells with mutant FLT3

Ultralight Structures for Space Solar Power Satellites

The design of a deployable spacecraft, measuring 60 m × 60 m, and with an areal density 100 g m^(−2) , is described. This spacecraft can be packaged into a cylinder measuring 1.5 m in height and 1 m in diameter. It can be deployed to a flat configuration, where it acts as a stiff, lightweight support framework for multifunctional tiles that collect sunlight, generate electric power, and transmit to a ground station on Earth

Early gene expression changes with rush immunotherapy

<p>Abstract</p> <p>Background</p> <p>To examine whether whole genome expression profiling could reveal changes in mRNA expression of peripheral blood mononuclear cells (PBMC) from allergic patients undergoing rush immunotherapy (RIT) that might be manifest within the first few months of treatment.</p> <p>Methods</p> <p>For this study, PBMC from three allergic patients undergoing RIT were assessed at four timepoints: prior to RIT, at 1 week and 7 week post-RIT, during build-up and at 4 months, after establishment of a maintenance dose. PBMC mRNA gene expression changes over time were determined by oligonucleotide microarrays using the Illumina Human-6 BeadChip Platform, which simultaneously interrogates expression profiles of > 47,000 transcripts. Differentially expressed genes were identified using well-established statistical analysis for microarrays. In addition, we analyzed peripheral blood basophil high-affinity IgE receptor (Fc epsilon RI) expression and T-regulatory cell frequency as detected by expression of CD3⁺CD4⁺CD25bright cells at each timepoint using flow cytometry.</p> <p>Results</p> <p>In comparing the initial 2 timepoints with the final 2 timepoints and analyzing for genes with ≥1.5-fold expression change (p less than or equal to 0.05, BH-FDR), we identified 507 transcripts. At a 2-fold change (p less than or equal to 0.05, BH-FDR), we found 44 transcripts. Of these, 28 were up-regulated and 16 were down-regulated genes. From these datasets, we have identified changes in immunologically relevant genes from both the innate and adaptive response with upregulation of expressed genes for molecules including IL-1β, IL-8, CD40L, BTK and BCL6. At the 4 month timepoint, we noted a downward trend in Fc epsilon RI expression in each of the three patients and increased allergen-specific IgG4 levels. No change was seen in the frequency of peripheral T-regulatory cells expressed over the four timepoints.</p> <p>Conclusions</p> <p>We observed significant changes in gene expression early in peripheral blood samples from allergic patients undergoing RIT. Moreover, serum levels for allergen specific IgG4 also increased over the course of treatment. These studies suggest that RIT induces rapid and dynamic alterations in both innate and adaptive immunity which can be observed in the periphery of allergic patients. These alterations could be directly related to the therapeutic shift in the allergen-specific class of immunoglobulin.</p

Tracking Antigen-Specific T-Cells during Clinical Tolerance Induction in Humans

Allergen immunotherapy presents an opportunity to define mechanisms of induction of clinical tolerance in humans. Significant progress has been made in our understanding of changes in T cell responses during immunotherapy, but existing work has largely been based on functional T cell assays. HLA-peptide-tetrameric complexes allow the tracking of antigen-specific T-cell populations based on the presence of specific T-cell receptors and when combined with functional assays allow a closer assessment of the potential roles of T-cell anergy and clonotype evolution. We sought to develop tools to facilitate tracking of antigen-specific T-cell populations during wasp-venom immunotherapy in people with wasp-venom allergy. We first defined dominant immunogenic regions within Ves v 5, a constituent of wasp venom that is known to represent a target antigen for T-cells. We next identified HLA-DRB1*1501 restricted epitopes and used HLA class II tetrameric complexes alongside cytokine responses to Ves v 5 to track T-cell responses during immunotherapy. In contrast to previous reports, we show that there was a significant initial induction of IL-4 producing antigen-specific T-cells within the first 3–5 weeks of immunotherapy which was followed by reduction of circulating effector antigen-specific T-cells despite escalation of wasp-venom dosage. However, there was sustained induction of IL-10-producing and FOXP3 positive antigen-specific T cells. We observed that these IL-10 producing cells could share a common precursor with IL-4-producing T cells specific for the same epitope. Clinical tolerance induction in humans is associated with dynamic changes in frequencies of antigen-specific T-cells, with a marked loss of IL-4-producing T-cells and the acquisition of IL-10-producing and FOXP3-positive antigen-specific CD4+ T-cells that can derive from a common shared precursor to pre-treatment effector T-cells. The development of new approaches to track antigen specific T-cell responses during immunotherapy can provide novel insights into mechanisms of tolerance induction in humans and identify new potential treatment targets

Identification of Critical Amino Acids in an Immunodominant IgE Epitope of Pen c 13, a Major Allergen from Penicillium citrinum

Background: Pen c 13, identified as a 33-kDa alkaline serine protease, is a major allergen secreted by Penicillium citrinum. Detailed knowledge about the epitopes responsible for IgE binding would help inform the diagnosis/prognosis of fungal allergy and facilitate the rational design of hypoallergenic candidate vaccines. The goal of the present study was to characterize the IgE epitopes of Pen c 13. Methodology/Principal Findings: Serum samples were collected from 10 patients with mold allergy and positive Pen c 13 skin test results. IgE-binding epitopes on rPen c 13 were mapped using an enzymatic digestion and chemical cleavage method, followed by dot-blotting and mass spectrometry. A B-cell epitope-predicting server and molecular modeling were used to predict the residues most likely involved in IgE binding. Theoretically predicted IgE-binding regions were further confirmed by site-directed mutagenesis assays. At least twelve different IgE-binding epitopes located throughout Pen c 13 were identified. Of these, peptides S16 (A 148 –E 166) and S22 (A 243 –K 274) were recognized by sera from 90 % and 100 % of the patients tested, and were further confirmed by inhibition assays. Peptide S22 was selected for further analysis of IgE-binding ability. The results of serum screening showed that the majority of IgE-binding ability resided in the C-terminus. One Pen c 13 mutant, G270A (T 261 –K 274), exhibited clearly enhanced IgE reactivity, whereas another, K274A, exhibited dramatically reduced IgE reactivity

Allergen-specific immunotherapy provides immediate, long-term and preventive clinical effects in children and adults: the effects of immunotherapy can be categorised by level of benefit -the centenary of allergen specific subcutaneous immunotherapy

Allergen Specific Immunotherapy (SIT) for respiratory allergic diseases is able to significantly improve symptoms as well as reduce the need for symptomatic medication, but SIT also has the capacity for long-term clinical effects and plays a protective role against the development of further allergies and symptoms. The treatment acts on basic immunological mechanisms, and has the potential to change the pathological allergic immune response. In this paper we discuss some of the most important achievements in the documentation of the benefits of immunotherapy, over the last 2 decades, which have marked a period of extensive research on the clinical effects and immunological background of the mechanisms involved. The outcome of immunotherapy is described as different levels of benefit from early reduction in symptoms over progressive clinical effects during treatment to long-term effects after discontinuation of the treatment and prevention of asthma. The efficacy of SIT increases the longer it is continued and immunological changes lead to potential long-term benefits. SIT alone and not the symptomatic treatment nor other avoidance measures has so far been documented as the therapy with long-term or preventive potential. The allergic condition is driven by a subset of T-helper lymphocytes (Th2), which are characterised by the production of cytokines like IL-4, and IL-5. Immunological changes following SIT lead to potential curative effects. One mechanism whereby immunotherapy suppresses the allergic response is through increased production of IgG4 antibodies. Induction of specific IgG4 is able to influence the allergic response in different ways and is related to immunological effector mechanisms, also responsible for the reduced late phase hyperreactivity and ongoing allergic inflammation. SIT is the only treatment which interferes with the basic pathophysiological mechanisms of the allergic disease, thereby creating the potential for changes in the long-term prognosis of respiratory allergy. SIT should not only be recognised as first-line therapeutic treatment for allergic rhinoconjunctivitis but also as secondary preventive treatment for respiratory allergic diseases

Prevalence of pre-frailty for the component of gait speed in older adults

OBJETIVO: investigar a pré-fragilidade e os fatores associados a essa condição, considerando as medidas de velocidade da marcha dos idosos. MÉTODO: a seleção dos participantes ocorreu por meio de critérios de inclusão/exclusão e teste de rastreamento cognitivo. A amostra foi calculada com base na estimativa da proporção populacional e constituída por 195 idosos, usuários de uma Unidade Básica de Saúde de Curitiba, PR. Os dados foram coletados mediante questionário sociodemográfico/clínico e teste de velocidade da marcha. RESULTADOS: a pré-fragilidade para velocidade da marcha possui moderada prevalência (27,3%) e associou-se à faixa etária entre 60 e 69 anos, baixa escolaridade, não se sentir solitário, utilizar anti-hipertensivo, apresentar doença cardiovascular e sobrepeso. CONCLUSÃO: considera-se relevante identificar os idosos na condição de pré-fragilidade, pois, dessa maneira, existe a possibilidade de intervenção imediata com a finalidade de estacionamento do quadro. É significativo o déficit de estudos sobre a síndrome da fragilidade em idosos brasileiros, principalmente aqueles que se referem a um componente isolado. Visto que a enfermagem gerontológica se encontra nos primeiros passos referentes à temática, entende-se que a identificação da prevalência deve ser o ponto primordial das pesquisas sobre o tema.Objetivo: investigar la prefragilidad y los factores asociados a esa condición, considerando medidas de velocidad de la marcha de los ancianos. Método: la selección de los participantes ocurrió por medio de criterios de inclusión/exclusión y prueba de rastreo cognitivo. La muestra fue calculada con base en la estimativa de la proporción poblacional y fue constituida por 195 ancianos usuarios de una Unidad Básica de Salud de Curitiba, PR. Los datos fueron recolectados mediante cuestionario sociodemográfico/clínico y prueba de velocidad de la marcha. Resultados: la prefragilidad para la velocidad de la marcha posee moderada prevalencia (27,3%) y se asoció al intervalo de edad entre 60 y 69 años, baja escolaridad, no sentirse solitario, utilizar antihipertensivos, presentar enfermedad cardiovascular y sobrepeso. Conclusión: se considera relevante identificar a los ancianos en la condición de prefragilidad, ya que de esa manera existe la posibilidad de intervenir inmediata con la finalidad de estacionar el cuadro. Es significativo el déficit de estudios sobre el síndrome de la fragilidad en ancianos brasileños, principalmente aquellos que se refieren a un componente aislado. Considerando que la enfermería gerontológica se encuentra en los primeros pasos en lo que se refiere a la temática, se entiende que la identificación de la prevalencia debe ser el punto primordial de las investigaciones sobre el tema.OBJECTIVE: to investigate pre-frailty and the factors associated with this condition, taking into account the measurements of the older adults' gait speed. METHOD: participants were selected by means of inclusion/exclusion criteria and a cognitive tracking test. The sample was calculated based on the estimation of populational proportion and made up of 195 older adults who were using a Primary Health-Care Center in Curitiba in the state of Paraná. Data was collected using a socio-demographic/clinical questionnaire and the gait speed test. RESULTS: pre-frailty for gait speed has moderate prevalence (27.3%), and is associated with the 60 - 69 years age range, a low level of schooling, not feeling oneself to be alone, using anti-hypertensives, having cardiovascular disease and being overweight. CONCLUSION: it is considered relevant to identify those older adults with pre-frailty, as this creates the possibility for immediate intervention with the aim of stabilizing the picture. There is a significant shortage of studies on the syndrome of frailty in Brazilian older adults, principally referring to components in isolation. Given that gerontological nursing is at an early stage regarding this issue, it is understood that the identification of the prevalence must be the key point of the research on the matter