114,687 research outputs found

    Clinical translation of [18F]ICMT-11 for measuring chemotherapy-induced caspase 3/7 activation in breast and lung cancer

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    Background: Effective anticancer therapy is thought to involve induction of tumour cell death through apoptosis and/or necrosis. [18F]ICMT-11, an isatin sulfonamide caspase-3/7-specific radiotracer, has been developed for PET imaging and shown to have favourable dosimetry, safety, and biodistribution. We report the translation of [18F]ICMT-11 PET to measure chemotherapy-induced caspase-3/7 activation in breast and lung cancer patients receiving first-line therapy. Results: Breast tumour SUVmax of [18F]ICMT-11 was low at baseline and unchanged following therapy. Measurement of M30/M60 cytokeratin-18 cleavage products showed that therapy was predominantly not apoptosis in nature. While increases in caspase-3 staining on breast histology were seen, post-treatment caspase-3 positivity values were only approximately 1%; this low level of caspase-3 could have limited sensitive detection by [18F]ICMT-11-PET. Fourteen out of 15 breast cancer patients responded to first–line chemotherapy (complete or partial response); one patient had stable disease. Four patients showed increases in regions of high tumour [18F]ICMT-11 intensity on voxel-wise analysis of tumour data (classed as PADS); response was not exclusive to patients with this phenotype. In patients with lung cancer, multi-parametric [18F]ICMT-11 PET and MRI (diffusion-weighted- and dynamic contrast enhanced-MRI) showed that PET changes were concordant with cell death in the absence of significant perfusion changes. Conclusion: This study highlights the potential use of [18F]ICMT-11 PET as a promising candidate for non-invasive imaging of caspase3/7 activation, and the difficulties encountered in assessing early-treatment responses. We summarize that tumour response could occur in the absence of predominant chemotherapy-induced caspase-3/7 activation measured non-invasively across entire tumour lesions in patients with breast and lung cancer

    Association of B-cell Lymphoma Protein-2 and Caspase-3 Expression in Ovarian Cancer

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    Ovarian cancer remains a major problem of women's health in the world, including Indonesia, and is associated with high rates of incidence and mortality. There are many efforts in early diagnosis on ovarian cancer, but until now there have not been found any satisfactory method. On the other hand, knowledge and research in the field of molecular biology become more advance, one of them is a mechanism to control the growth of cells in ovarian cancer through a process of programmed cell death or apoptosis. B-cell lymphoma protein 2 (Bcl-2) and caspase-3 are proteins that play a role on the mechanism of apoptosis. The purpose of this study was to determine the expression of Bcl-2 and caspase-3 and their association with ovarian cancer. Materials and method: The design of this study was a cross-sectional study. Expression of Bcl-2 and caspase-3 examined by immunohistochemistry under light microscope with 400x light power field and expression as a negative when the protein expressed in 10% or less of cells and as a positive when the protein expressed in more than 10% of cells. A number of 45 subjects were recruited in this study. Thirthy one of 45 subjects showed the expression of Bcl-2 positive (68.9%), while the positive expression of caspase-3 present in 20 subjects (44.4%). There was a significant association between the expression of Bcl-2 with the expression of caspase-3 in ovarian cancer patients (p=0.002; lambda=0.4). There was also a significant association between stage of disease with expression of Bcl-2 (p=0.002; lambda=0.3) dan expression of caspase-3 (p=0.001; lambda=0.3). Conclusion: It concluded that there is a significant association between the expression of Bcl-2 and the expression of caspase-3 in ovarian cancer

    Combined bezafibrate and medroxyprogesterone acetate: potential novel therapy for acute myeloid leukaemia

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    Background: The majority of acute myeloid leukaemia (AML) patients are over sixty years of age. With current treatment regimens, survival rates amongst these, and also those younger patients who relapse, remain dismal and novel therapies are urgently required. In particular, therapies that have anti-leukaemic activity but that, unlike conventional chemotherapy, do not impair normal haemopoiesis. Principal Findings: Here we demonstrate the potent anti-leukaemic activity of the combination of the lipid-regulating drug bezafibrate (BEZ) and the sex hormone medroxyprogesterone acetate (MPA) against AML cell lines and primary AML cells. The combined activity of BEZ and MPA (B/M) converged upon the increased synthesis and reduced metabolism of prostaglandin D2 (PGD2) resulting in elevated levels of the downstream highly bioactive, anti-neoplastic prostaglandin 15-deoxy Δ12,14 PGJ2 (15d-PGJ2). BEZ increased PGD2 synthesis via the generation of reactive oxygen species (ROS) and activation of the lipid peroxidation pathway. MPA directed prostaglandin synthesis towards 15d-PGJ2 by inhibiting the PGD2 11β -ketoreductase activity of the aldo-keto reductase AKR1C3, which metabolises PGD2 to 9α11β-PGF2α. B/M treatment resulted in growth arrest, apoptosis and cell differentiation in both AML cell lines and primary AML cells and these actions were recapitulated by treatment with 15d-PGJ2. Importantly, the actions of B/M had little effect on the survival of normal adult myeloid progenitors. Significance: Collectively our data demonstrate that B/M treatment of AML cells elevated ROS and delivered the anti-neoplastic actions of 15d-PGJ2. These observations provide the mechanistic rationale for the redeployment of B/M in elderly and relapsed AML

    Transcriptome dynamics in the asexual cycle of the chordate Botryllus schlosseri

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    Background: We performed an analysis of the transcriptome during the blastogenesis of the chordate Botryllus schlosseri, focusing in particular on genes involved in cell death by apoptosis. The tunicate B. schlosseri is an ascidian forming colonies characterized by the coexistence of three blastogenetic generations: filter-feeding adults, buds on adults, and budlets on buds. Cyclically, adult tissues undergo apoptosis and are progressively resorbed and replaced by their buds originated by asexual reproduction. This is a feature of colonial tunicates, the only known chordates that can reproduce asexually. Results: Thanks to a newly developed web-based platform (http://botryllus.cribi.unipd.it), we compared the transcriptomes of the mid-cycle, the pre-take-over, and the take-over phases of the colonial blastogenetic cycle. The platform is equipped with programs for comparative analysis and allows to select the statistical stringency. We enriched the genome annotation with 11,337 new genes; 581 transcripts were resolved as complete open reading frames, translated in silico into amino acid sequences and then aligned onto the non-redundant sequence database. Significant differentially expressed genes were classified within the gene ontology categories. Among them, we recognized genes involved in apoptosis activation, de-activation, and regulation. Conclusions: With the current work, we contributed to the improvement of the first released B. schlosseri genome assembly and offer an overview of the transcriptome changes during the blastogenetic cycle, showing up- and down-regulated genes. These results are important for the comprehension of the events underlying colony growth and regression, cell proliferation, colony homeostasis, and competition among different generations

    Relevance of the variability of the feline immunodeficiency virus in regard to pathogenicity and vaccination in New Zealand : a thesis presented in partial fulfilment of the requirements for the degree of Doctor of Philosophy in Animal Science, Massey University, Manawatū, New Zealand

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    Cats infected with the feline immunodeficiency virus (FIV) show a range of clinical signs. Given the variability of the FIV genome, it is possible that there is variation in certain biological characteristics of FIV, such as pathogenicity. This may also be relevant to vaccination against FIV, as an effective vaccine would have to result in the generation of T cells that recognise a range of different variants in the field. The Fel-O-Vax® FIV vaccine has been available to veterinarians in New Zealand (NZ) for the past 12 years. Despite this, there is a paucity of studies investigating the cross-reactivity of the vaccine-induced immune response against different variants of FIV, and no studies investigating the efficacy of the vaccine in NZ. The overall aim of the research in this thesis was to determine the relevance of the variability of FIV, in regard to pathogenicity and vaccination in NZ. Firstly, 2 separate assays were designed to assess variation in the ability of different isolates of FIV to induce apoptosis or inhibit mitogen-induced proliferation in lymphoid cells in vitro. Results showed that variation in FIV-apoptosis did occur, supporting the argument that FIV variants may also differ in pathogenicity. Secondly, the cross-reactivity of the vaccine-induced immune response was assessed in vitro and in vivo, by measuring antigen-specific cellular activation and a delayed type hypersensitivity (DTH) response in vaccinated cats following inoculation with NZ field isolates of FIV. Results showed that the response was at least partially cross-reactive, however quantitative differences were detected in the response to each isolate of FIV tested. Finally, efficacy of the Fel-O-Vax® FIV vaccine under NZ conditions was investigated by comparing the prevalence of FIV in vaccinated and unvaccinated cats in the field. Results showed that there was no effect of vaccination on FIV prevalence, suggesting poor efficacy of the Fel-O-Vax® FIV vaccine in NZ. Results described in this thesis support the argument that there is variation among FIV in NZ, and that this may affect pathogenicity and vaccine efficacy in this country. The evidence presented did not support use of the Fel-O-Vax® FIV vaccine in NZ

    Bayesian inference and model choice in a hidden stochastic two-compartment model of hematopoietic stem cell fate decisions

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    Despite rapid advances in experimental cell biology, the in vivo behavior of hematopoietic stem cells (HSC) cannot be directly observed and measured. Previously we modeled feline hematopoiesis using a two-compartment hidden Markov process that had birth and emigration events in the first compartment. Here we perform Bayesian statistical inference on models which contain two additional events in the first compartment in order to determine if HSC fate decisions are linked to cell division or occur independently. Pareto Optimal Model Assessment approach is used to cross check the estimates from Bayesian inference. Our results show that HSC must divide symmetrically (i.e., produce two HSC daughter cells) in order to maintain hematopoiesis. We then demonstrate that the augmented model that adds asymmetric division events provides a better fit to the competitive transplantation data, and we thus provide evidence that HSC fate determination in vivo occurs both in association with cell division and at a separate point in time. Last we show that assuming each cat has a unique set of parameters leads to either a significant decrease or a nonsignificant increase in model fit, suggesting that the kinetic parameters for HSC are not unique attributes of individual animals, but shared within a species.Comment: Published in at http://dx.doi.org/10.1214/09-AOAS269 the Annals of Applied Statistics (http://www.imstat.org/aoas/) by the Institute of Mathematical Statistics (http://www.imstat.org

    Genome Wide Transcriptome Analysis of Dendritic Cells Identifies Genes with Altered Expression in Psoriasis

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    Activation of dendritic cells by different pathogens induces the secretion of proinflammatory mediators resulting in local inflammation. Importantly, innate immunity must be properly controlled, as its continuous activation leads to the development of chronic inflammatory diseases such as psoriasis. Lipopolysaccharide (LPS) or peptidoglycan (PGN) induced tolerance, a phenomenon of transient unresponsiveness of cells to repeated or prolonged stimulation, proved valuable model for the study of chronic inflammation. Thus, the aim of this study was the identification of the transcriptional diversity of primary human immature dendritic cells (iDCs) upon PGN induced tolerance. Using SAGESeq approach, a tag-based transcriptome sequencing method, we investigated gene expression changes of primary human iDCs upon stimulation or restimulation with Staphylococcus aureus derived PGN, a widely used TLR2 ligand. Based on the expression pattern of the altered genes, we identified non-tolerizeable and tolerizeable genes. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (Kegg) analysis showed marked enrichment of immune-, cell cycle- and apoptosis related genes. In parallel to the marked induction of proinflammatory mediators, negative feedback regulators of innate immunity, such as TNFAIP3, TNFAIP8, Tyro3 and Mer are markedly downregulated in tolerant cells. We also demonstrate, that the expression pattern of TNFAIP3 and TNFAIP8 is altered in both lesional, and non-lesional skin of psoriatic patients. Finally, we show that pretreatment of immature dendritic cells with anti-TNF-α inhibits the expression of IL-6 and CCL1 in tolerant iDCs and partially releases the suppression of TNFAIP8. Our findings suggest that after PGN stimulation/restimulation the host cell utilizes different mechanisms in order to maintain critical balance between inflammation and tolerance. Importantly, the transcriptome sequencing of stimulated/restimulated iDCs identified numerous genes with altered expression to date not associated with role in chronic inflammation, underlying the relevance of our in vitro model for further characterization of IFNprimed iDCs

    Effect of tibolone on breast cancer cell proliferation in postmenopausal ER+ patients: Results from STEM trial

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    Purpose: Tibolone is a selective tissue estrogenic activity regulator, approved for the treatment of vasomotor symptoms in postmenopausal women. We have done an exploratory, double-blind, randomized, placebo-controlled pilot trial to investigate the tissue-specific effects of 2.5 mg tibolone on breast cancer in postmenopausal women, in particular on tissue proliferation (STEM, Study of Tibolone Effects on Mamma carcinoma tissue). Experimental Design: Postmenopausal women with initially stage I/II, estrogen receptor-positive (ER+) primary breast cancer, were randomly assigned to 14 days of placebo or 2.5 mg/d tibolone. Core biopsies of the primary tumor were obtained before and after treatment. Ki-67 and apoptosis index were analyzed in baseline and corresponding posttreatment specimen. Results: Of 102 enrolled patients, 95 had evaluable data. Baseline characteristics were comparable between both treatment groups. Breast cancer cases are mainly invasive (99%), stage I or II (42% and 50% respectively), and ER+ (99%). Median intratumoral Ki-67 expression at baseline was 13.0%, in the tibolone group and 17.8% in the placebo group, and decreased to 12.0% after 14 days of tibolone while increasing to 19.0% in the placebo group. This change from baseline was not significantly different between tibolone and placebo (Wilcoxon test; P = 0.17). A significant difference was observed between the treatment groups when the median change from baseline apoptosis index was compared between the treatment groups (tibolone, 0.0%; placebo, +0.3%; Wilcoxon test; P = 0.031). The incidence of adverse effects was comparable. Conclusions: In ER+ breast tumors, 2.5 mg/d tibolone given for 14 days has no significant effect on tumor cell proliferation
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