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1.
Hum Immunol ; 80(4): 257-262, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30790598

RESUMO

INTRODUCTION: Chromosomal abnormalities are frequent events in hematological malignancies. The degree of HLA compatibility between donor and recipient in hematopoietic stem cell transplantation is critical. PURPOSE OF THE STUDY: In this report, we describe an acute myeloid leukemia case with loss of heterozygosity (LOH) encompassing the entire HLA. MATERIALS AND METHODS: HLA molecular typing was performed on peripheral blood (PB) and buccal swabs (BS). Chromosomal microarray analysis (CMA) was performed using a whole genome platform. RESULTS: Typing results on PB sample collected during blast crisis demonstrated homozygosity at the -A, -B, -C, -DR, and -DQ loci. A BS sample demonstrated heterozygosity at all loci. A subsequent PB sample drawn after count recovery confirmed heterozygosity. The CMA performed on PB samples collected during and after blast crisis revealed a large terminal region of copy-neutral LOH involving chromosome region 6p25.3p21.31, spanning approximately 35.9 Mb. The results of the CMA assay on sample collected after count recovery did not demonstrate LOH. CONCLUSIONS: LOH at the HLA gene locus may significantly influence the donor search resulting in mistakenly choosing homozygous donors. We recommend confirming the HLA typing of recipients with hematological malignancies when homozygosity is detected at any locus by using BS samples, or alternatively from PB when remission is achieved.


Assuntos
Medula Óssea/fisiologia , Genoma/genética , Transplante de Células-Tronco Hematopoéticas , Leucemia Mieloide Aguda/genética , Leucócitos Mononucleares/fisiologia , Perda de Heterozigosidade , Complexo Principal de Histocompatibilidade/genética , Idoso , Circulação Sanguínea , Feminino , Teste de Histocompatibilidade , Humanos , Leucemia Mieloide Aguda/diagnóstico , Leucemia Mieloide Aguda/terapia , Análise em Microsséries , Técnicas de Diagnóstico Molecular , Indução de Remissão
2.
Sci Rep ; 5: 16595, 2015 Nov 13.
Artigo em Inglês | MEDLINE | ID: mdl-26563826

RESUMO

Retinal ganglion cell (RGC) injury and cell death from glaucoma and other forms of optic nerve disease is a major cause of irreversible vision loss and blindness. Human pluripotent stem cell (hPSC)-derived RGCs could provide a source of cells for the development of novel therapeutic molecules as well as for potential cell-based therapies. In addition, such cells could provide insights into human RGC development, gene regulation, and neuronal biology. Here, we report a simple, adherent cell culture protocol for differentiation of hPSCs to RGCs using a CRISPR-engineered RGC fluorescent reporter stem cell line. Fluorescence-activated cell sorting of the differentiated cultures yields a highly purified population of cells that express a range of RGC-enriched markers and exhibit morphological and physiological properties typical of RGCs. Additionally, we demonstrate that aligned nanofiber matrices can be used to guide the axonal outgrowth of hPSC-derived RGCs for in vitro optic nerve-like modeling. Lastly, using this protocol we identified forskolin as a potent promoter of RGC differentiation.


Assuntos
Sistemas CRISPR-Cas/genética , Diferenciação Celular/genética , Células-Tronco Embrionárias/metabolismo , Engenharia Genética/métodos , Células Ganglionares da Retina/metabolismo , Animais , Linhagem Celular , Células Cultivadas , Células-Tronco Embrionárias/citologia , Expressão Gênica , Humanos , Imuno-Histoquímica , Potenciais da Membrana/genética , Camundongos , Microscopia de Fluorescência , Células Ganglionares da Retina/citologia , Células Ganglionares da Retina/fisiologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Antígenos Thy-1/metabolismo , Fatores de Tempo , Fator de Transcrição Brn-3B/genética , Fator de Transcrição Brn-3B/metabolismo
3.
Hum Vaccin ; 5(5): 322-31, 2009 May.
Artigo em Inglês | MEDLINE | ID: mdl-19221516

RESUMO

Development of optimal formulation conditions stabilizing live attenuated bacterial vaccines is impeded by traditional methods used for viability measurement. To facilitate preformulation studies of such vaccines, spectroscopic techniques capable of providing real-time and high throughput information have been employed to obtain a global stability profile for a live attenuated Ty21a bacterial typhoid vaccine over a wide range of pH (4 to 8) and temperature (10 to 85 degrees C). Using the data obtained from fluorescence and circular dichroism techniques, an empirical phase diagram (EPD) has been subsequently constructed, which suggests that Ty21a cells exist in at least four apparent physical phases related to different viability states, with the most stable phase at pH 6 and 7 at temperatures below 30 degrees C. A slightly basic pH (pH 8) appears to decrease the fluidity of the cell membrane, whereas acidic pH conditions are detrimental to membrane integrity over the entire temperature range. Based on the above stability profile, a fluorescence-based high throughput screening assay has been developed to test the stabilizing effects of various compounds at different concentrations. Amongst other promising stabilizers, 10% sucrose and 0.15 M glutamic acid display the greatest protective effects, with an increase of about 10 degrees C in the transition temperature of Ty21a cells. Preliminary studies have also been performed on foam dried formulations as an alternative approach to further stabilize Ty21a cells. The data show that 10% sucrose and trehalose both increase the in-process and storage stabilities of the cells.


Assuntos
Viabilidade Microbiana , Polissacarídeos Bacterianos/imunologia , Vacinas Tíficas-Paratíficas/imunologia , Dicroísmo Circular , Estabilidade de Medicamentos , Excipientes/farmacologia , Ácido Glutâmico/farmacologia , Humanos , Concentração de Íons de Hidrogênio , Polissacarídeos Bacterianos/genética , Análise Espectral/métodos , Sacarose/farmacologia , Temperatura , Vacinas Tíficas-Paratíficas/genética , Vacinas Atenuadas/genética , Vacinas Atenuadas/imunologia
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