Identificaçao e caracterizaçao do gene rpoC de Herbaspirillum seropedicae

Orientador: Emanuel M. de SouzaMonografia (Bacharelado) - Universidade Federal do Paraná. Setor de Ciencias Biológicas. Curso de Graduaçao em Ciencias BiológicasResumo : A regulação da expressão gênica ocorre principalmente a nível transcricional. O principal alvo deste controle, em bactérias, é a RNA polimerase DNAdependente em resposta a fatores intrínsecos e extrínsecos. A regulação da transcrição está mais bem caracterizada em E. coli. A RNA polimerase de E. co/i é uma enzima oligomérica que possuí duas subunidades a, uma ~, uma W e uma 0). A subunidade W é responsável pela ligação ao DNA molde e pela interação com os fatores cr. A região C-terminal da subunidade W não está envolvida com a atividade catalítica ou com a montagem da holoenzima. Poucos estudos sobre a RNA polimerase de outros organismos foram feitos. Neste trabalho, o gene rpoC de Herbaspirillum seropedícae, que codifica para a subunidade W da RNA polimerase, foi identificada no Banco de Dados do Programa GENOPAR. Este gene foi amplificado por PCR usando oligonucleotídeos iniciadores complementares a seqüência das regiões que codificam para a extremidade N-terminal e C-terminal da subunídade W da RNA polimerase de H. seropedicae. As condições ótimas de amplificação foram determinadas. O fragmento obtido possui 4242pb e está sendo clonado no vetor pET29a. O clone resultante deverá expressar a subunidade W ligada a uma cauda de histidina

Refractory Angina Cell Therapy (ReACT) Involving Autologous Bone Marrow Cells in Patients Without Left Ventricular Dysfunction: A Possible Role for Monocytes

Autologous bone marrow mononuclear cell (BMMC) transplantation has emerged as a potential therapeutic option for refractory angina patients. Previous studies have shown conflicting myocardium reperfusion results. the present study evaluated safety and efficacy of CellPraxis Refractory Angina Cell Therapy Protocol (ReACT). in which a specific BMMC formulation was administered as the sole therapy for these patients. the phase I/IIa noncontrolled, open label. clinical trial, involved eight patients with refractory angina and viable ischemic myocardium, without left ventricular dysfunction and who were not suitable for conventional myocardial revascularization. ReACT is a surgical procedure involving a single series of multiple injections (40-90 injections, 0.2 ml each) into ischemic areas of the left ventricle. Primary endpoints were Canadian Cardiovascular Society Angina Classification (CCSAC) improvement at 18 months follow-up and myocardium ischemic area reduction (assessed by scintigraphic analysis) at 12 months follow-up, in correlation with a specific BMMC formulation. Almost all patients presented progressive improvement in angina classification beginning 3 months (p = 0.008) postprocedure which was sustained at 18 months follow-up (p = 0.004), as well as objective myocardium ischemic area reduction at 12 months (decrease of 84.4%, p < 0.004). A positive correlation was found between monocyte concentration and CCSAC improvement (r = -0.759, p < 0.05). Improvement in CCSAC, followed by correlated reduction in scintigraphic myocardium ischemic area, strongly suggests neoangiogenesis as the main stem cell action mechanism. the significant correlation between number of monocytes and improvement strongly supports a cell-related effect of ReACT. ReACT appeared safe and effective.Cryopraxis Crobiologia Ltda.Cellpraxis BiogenhariaUniversidade Federal de São Paulo, Paulista Sch Med, Dept Surg, Div Cardiovasc Surg, São Paulo, BrazilCryopraxis Criobiol Ltda, Rio de Janeiro, BrazilCellpraxis Bioengn, Rio de Janeiro, BrazilUniversidade Federal de São Paulo, Paulista Sch Med, Div Hematol, São Paulo, BrazilUniv São Paulo, Coll Med, Inst Heart, São Paulo, BrazilUniv S Florida, Coll Med, Dept Neurosurg & Brain Repair, Ctr Excellence Aging & Brain Repair, Tampa, FL USAUniv S Florida, Off Res & Innovat, Tampa, FL USAUniversidade Federal de São Paulo, Paulista Sch Med, Dept Surg, Div Cardiovasc Surg, São Paulo, BrazilUniversidade Federal de São Paulo, Paulista Sch Med, Div Hematol, São Paulo, BrazilWeb of Scienc

Determinaçăo de seqüęncias de DNA reconhecidas por proteínas reguladoras de transcriçăo dependentes do fator sigma 54 da RNA-polimerase de Herbaspirillum seropedicae /

Banca: Emanuel Maltempi de Souza [Orientador]Data da defesa: 2006.02.27Banca: Fábio de Oliveira Pedrosa [Orientador]Dissertaçăo (mestrado) - Universidade Federal do Paraná, Setor de Cięncias Biológicas, Programa de Pós-Graduaçăo em Bioquímica. Defesa: Curitiba, 27/02/2006Inclui bibliografi

Caracterização molecular e funcional das células do endométrio de mulheres com endometriose pélvica

The endometrium contains stem cells that possibly contribute to the regeneration of the functional layer during the mensal cycling. Different kinds of stem cells were isolated from endometrium and endometriotic implants. The monoclonal origin of endometriotic implants indicates that stem cells may have a role in the endometriosis pathogenesis. We isolated a mixed cells pool containing at least one stem cells population from healthy and endometriosis endometrium. These cells presented the markers CD90, CD73, CD105, CD44 and CD146, being able to differentiate into adipogenic, osteogenic and chondrogenic lineages and not responding to progesterone stimulus. The endometriosis stem cells expressed the CD34 naïve stem cells marker, presented higher proliferative rates than healthy ones and were able to form long lasting cells aggregates (spheroids) in tridimensional cultures. The endometrium with endometriosis had a unique stem cell population, reinforcing the hypothesis that stem cells play a key role in the endometriosis pathogenesis. 2. INTRODUCTION Stem cells are undifferentiated cells defined by their functional ability to self-renew and to differentiate into multiple cell lineages, as well as by plasticity and clonogenic potential. They were found abundantly in embryonic and fetal tissues and are also present in small amounts in the majority of the adult tissues; differing between each other only in the clonogenic and differentiation potential. Adult stem cells occur in niches that balance self-renewal with lineage selection and progression during tissue homeostasisEndometriose é uma doença ginecológica crônica, benigna, caracterizada pela presença de tecido endometrial fora do útero. Estima-se que 10% a 15% das mulheres em idade reprodutiva padeçam da afecção e a imensa maioria experimenta diminuição importante na qualidade de vida. Dentre os sintomas, destacam-se a dismenorreia, dispareunia e infertilidade. Dentre os diversos achados acerca da patogenia da endometriose, cumpre ressaltar que a maior proliferação do tecido endometrial, inclusive ectópico, foi demonstrada por diversos estudos. Neste trabalho foi realizado o isolamento e caracterização fisiológica e fenotípica das células-tronco derivadas do endométrio de mulheres com endometriose (EeSCs). Essas células apresentaram alta expressão do marcador CD34, bem como expressaram também o marcador mesenquimal CD146. Elas mostraram-se capazes de diferenciar-se em três linhagens mesenquimais: adipócitos, condrócitos e osteócitos. Além disso, possuem altas taxas de proliferação celular, mesmo na presença de progesterona, e secretam grandes concentrações de IL-6 e IL-8 quando cultivadas isoladamente, e IL-6, IL-8 e IL-1? quando co-cultivadas com células-tronco obtidas de endométrio saudável. As EeSCs possuem a habilidade de formar esferoides celulares quando cultivadas em condições que mimetizem o ambiente peritoneal. Esses dados demonstram que possivelmente o endométrio de mulheres com endometriose possui uma população específica de células-tronco que deve estar diretamente envolvida na etiologia/fisiopatologia da doença.Dados abertos - Sucupira - Teses e dissertações (2013 a 2016

Determinaçăo de seqüęncias de DNA reconhecidas por proteínas reguladoras de transcriçăo dependentes do fator sigma 54 da RNA-polimerase de Herbaspirillum seropedicae /

Banca: Emanuel Maltempi de Souza [Orientador]Data da defesa: 2006.02.27Banca: Fábio de Oliveira Pedrosa [Orientador]Dissertaçăo (mestrado) - Universidade Federal do Paraná, Setor de Cięncias Biológicas, Programa de Pós-Graduaçăo em Bioquímica. Defesa: Curitiba, 27/02/2006Inclui bibliografi

Overhauling CAR T Cells to Improve Efficacy, Safety and Cost

Gene therapy is now surpassing 30 years of clinical experience and in that time a variety of approaches has been applied for the treatment of a wide range of pathologies. While the promise of gene therapy was over-stated in the 1990&rsquo;s, the following decades were met with polar extremes between demonstrable success and devastating setbacks. Currently, the field of gene therapy is enjoying the rewards of overcoming the hurdles that come with turning new ideas into safe and reliable treatments, including for cancer. Among these modalities, the modification of T cells with chimeric antigen receptors (CAR-T cells) has met with clear success and holds great promise for the future treatment of cancer. We detail a series of considerations for the improvement of the CAR-T cell approach, including the design of the CAR, routes of gene transfer, introduction of CARs in natural killer and other cell types, combining the CAR approach with checkpoint blockade or oncolytic viruses, improving pre-clinical models as well as means for reducing cost and, thus, making this technology more widely available. While CAR-T cells serve as a prime example of translating novel ideas into effective treatments, certainly the lessons learned will serve to accelerate the current and future development of gene therapy drugs