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1.
J Immigr Minor Health ; 23(5): 1011-1020, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-33043411

RESUMO

African American and Hispanic women report less physical activity (PA) than non-Hispanic White women. As such, a digitally-enhanced 16-week social support pilot intervention was conducted to promote PA among African American and Hispanic women dyads. This study quantitatively and qualitatively examined the engagement and satisfaction of participants (N = 30; 15 dyads) assigned to the intervention. Intervention participants received telephone counseling calls based on motivational interviewing and a Jawbone UP activity monitor. Intervention engagement and satisfaction data were collected from the Jawbone UP, call logs, self-report questionnaires conducted at the 16-week follow-up, and two post-intervention focus groups. Nonparametric tests assessed group differences across engagement and satisfaction measures, and a manually-driven coding scheme was used to evaluate emerging themes from qualitative text. Participants demonstrated high engagement in the telephone counseling sessions and moderate engagement with the Jawbone UP. Friend/co-worker dyads and participants who were 45 years and older were more likely to use the device. Qualitative results emphasized participants' appreciation for the counseling calls, the Jawbone UP, and the overall dyadic framework of the study to collectively nurture social support and accountability for PA. Overall, the intervention group reacted positively to study components. Additional research is needed to understand the role of technology in facilitating long-lasting PA change via social support in minority populations.


Assuntos
Negro ou Afro-Americano , Satisfação Pessoal , Exercício Físico , Feminino , Humanos , Projetos Piloto , Apoio Social
2.
Gene ; 210(1): 103-8, 1998 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-9524237

RESUMO

Caprine interferon-gamma (IFN-gamma) cDNA was cloned from mitogen stimulated peripheral blood mononuclear cell (PBMC) RNA utilizing the reverse transcription-polymerase chain reaction (RT-PCR). The cDNA open reading frame (ORF) is 498bp, encoding a putative 166 amino acid (aa) protein (19327Da). The predicted aa sequence homology of caprine IFN-gamma and the corresponding ovine, bovine and cervine cytokine is 98.8%, 95.2% and 92.8%, respectively. IFN-gamma cDNA was subcloned and expressed in two different plasmids under the control of either the human cytomegalovirus (CMV) immediate early promoter or the caprine arthritis-encephalitis virus long terminal repeat (CAEV LTR). Recombinant caprine IFN-gamma (rCaIFN-gamma) secreted by transfected COS-7 cells shared at least two antigenic epitopes with recombinant bovine IFN-gamma (rBoIFN-gamma) and exhibited biological activity in the vesicular stomatitis virus (VSV) cytopathic effect reduction assay. In-vivo expression of IFN-gamma cDNA promoted by the CAEV LTR was confirmed by the intramuscular (IM) injection of Balb/C mice with plasmid followed by Western blot analysis of mouse serum against purified rCaIFN-gamma produced in E. coli.


Assuntos
Interferon gama/química , Leucócitos Mononucleares/química , Sequência de Aminoácidos , Animais , Sequência de Bases , Células COS , Clonagem Molecular , Escherichia coli/genética , Cabras , Interferon gama/farmacologia , Camundongos , Camundongos Endogâmicos BALB C , Dados de Sequência Molecular , Fases de Leitura Aberta/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/farmacologia , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Transfecção/genética , Vírus da Estomatite Vesicular Indiana/efeitos dos fármacos , Ensaio de Placa Viral
3.
Cell Growth Differ ; 8(12): 1371-80, 1997 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-9419425

RESUMO

Expression of Pim-1, an oncogenic serine/threonine kinase, is highly regulated at the transcriptional, posttranscriptional, and posttranslational levels. Here, we report that expression of Pim-1 kinase is additionally regulated at the translational level. Pim-1 protein expression did not increase in Hut-78 lymphocytes in response to PMA1/ionomycin stimulation despite approximately 20-fold increases in mRNA levels, suggesting that translation was repressed. Sequence analysis of the 5'-untranslated region (UTR) indicated a long (400 nucleotide), 76% G + C-rich region, characteristics known to inhibit translation. Deletion of the 5'-UTR of pim-1 increased translation of the Pim-1 protein approximately 10-fold in vitro in reticulocyte lysates and approximately 1.6-fold in vivo in NIH-3T3 cells. When full-length 5'-UTR-containing pim-1 cDNA constructs were transfected into NIH-3T3 cells overexpressing eukaryotic translation initiation factor 4E (eIF-4E), approximately 6-fold higher levels of Pim-1 protein were produced, as compared to that produced in control NIH-3T3 cells. Moreover, eIF-4E overexpression had little effect in the absence of the 5'-UTR, suggesting that it relieved 5'-UTR-mediated inhibition of Pim-1 expression.


Assuntos
Conformação de Ácido Nucleico , Fatores de Iniciação de Peptídeos/fisiologia , Proteínas Serina-Treonina Quinases/metabolismo , Proteínas Proto-Oncogênicas/biossíntese , RNA Mensageiro/fisiologia , Células 3T3 , Animais , Sequência de Bases , Linhagem Celular , Fator de Iniciação 4E em Eucariotos , Humanos , Metionina/metabolismo , Camundongos , Dados de Sequência Molecular , Nigericina/análogos & derivados , Nigericina/farmacologia , Fatores de Iniciação de Peptídeos/biossíntese , Proteínas Proto-Oncogênicas/genética , Proteínas Proto-Oncogênicas c-pim-1 , RNA Mensageiro/análise , Reticulócitos/metabolismo , Análise de Sequência de RNA , Acetato de Tetradecanoilforbol/análogos & derivados , Acetato de Tetradecanoilforbol/farmacologia , Transcrição Gênica , Transfecção , beta-Galactosidase/análise
4.
Endocrinology ; 136(12): 5252-9, 1995 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-7588268

RESUMO

The lactogen-dependent Nb2 lymphoma line (Nb2-11) represents a useful pre-T cell model for investigation of early molecular events coupled to PRL-stimulated cell cycle progression. Expression of pim-1, a protooncogene that encodes a conserved cytosolic serine/threonine protein kinase, is rapidly induced in hematopoietic cells upon mitogen stimulation and is thought to be important for lymphocyte activation. The present study was conducted to determine whether mitogen stimulation in Nb2-11 or lactogen-independent Nb2-SFJCD1 cells provokes pim-1 gene expression. The pim-1 transcript was undetectable in control growth-arrested Nb2-11 cultures; however, PRL rapidly stimulated its expression in a biphasic manner. Peak expression occurred within 2-4 h (> 40-fold) and was followed by a second elevation at 12 h. The effect of PRL and IL-2 to induce pim-1 at 2 h was concentration dependent and not inhibited by cycloheximide. In Nb2-SFJCD1 cells, pim-1 messenger RNA was expressed in control cultures and augmented by PRL stimulation. Results from stability studies indicated that the t1/2 values for the pim-1 transcript were 79 and 81 min in PRL-stimulated Nb2-11 cells at 2 and 12 h. However, in the lactogen-treated Nb2-SFJCD1 line, it was nearly 3-fold more stable (219 min) at 2 h compared to that determined at either 12 h or in unstimulated cultures. In other experiments, PRL-stimulated expression of the pim-1 protein was evaluated in [35S]methionine-labeled cells by immunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. In Nb2-11 cells, enhanced [35S]pim-1 expression paralleled its messenger RNA transcription through 8 h. Elevated [35S]pim-1 was detected within 1 h and peaked by 2-4 h. Therefore, pim-1 represents an immediate early gene induced by PRL stimulation in Nb2-11 cells. Its initial peak of transcription occurs early during G1 cell cycle progression, whereas a second elevation is coincident with the G1/S transition. These results demonstrate that mitogen-induced expression of pim-1 is a rapid event in Nb2 lymphoma cells and suggest that it may be associated with cell cycle progression.


Assuntos
Regulação da Expressão Gênica/efeitos dos fármacos , Interleucina-2/farmacologia , Prolactina/farmacologia , Proteínas Serina-Treonina Quinases , Proteínas Proto-Oncogênicas/genética , Proto-Oncogenes/efeitos dos fármacos , Animais , Ciclo Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Linfoma/genética , Inibidores da Síntese de Proteínas/farmacologia , Proteínas Proto-Oncogênicas/análise , Proteínas Proto-Oncogênicas c-pim-1 , Ratos , Transcrição Gênica/efeitos dos fármacos , Células Tumorais Cultivadas
5.
Arch Biochem Biophys ; 298(2): 594-601, 1992 Nov 01.
Artigo em Inglês | MEDLINE | ID: mdl-1416988

RESUMO

The human pim-1 proto-oncogene was expressed in Escherichia coli as a glutathione-S-transferase (GST)-fusion protein and the enzymatic properties of its kinase activity were characterized. Likewise, a Pim-1 mutant lacking intrinsic kinase activity was constructed by site-directed mutagenesis (Lys67 to Met) and expressed in E. coli. In vitro assays with the mutant Pim-1 kinase showed no contaminating kinase activity. The wild-type Pim-1 kinase-GST fusion protein showed a pH optimum of 7 to 7.5 and optimal activity was observed at either 10 mM MgCl2 or 5 mM MnCl2. Higher cation concentrations were inhibitory, as was the addition of NaCl to the assays. Previous work by this laboratory assaying several proteins and peptides showed histone H1 and the peptide Kemptide to be efficiently phosphorylated by recombinant Pim-1 kinase. Here we examine the substrate sequence specificity of Pim-1 kinase in detail. Comparison of different synthetic peptide substrates showed Pim-1 to have a strong substrate preference for the peptide Lys-Arg-Arg-Ala-Ser*-Gly-Pro with an almost sixfold higher specificity constant kcat/Km over that of the substrate Kemptide (Leu-Arg-Arg-Ala-Ser*-Leu-Gly). The presence of basic amino acid residues on the amino terminal side of the target Ser/Thr was shown to be essential for peptide substrate recognition. Furthermore, phosphopeptide analysis of calf thymus histone H1 phosphorylated in vitro by Pim-1 kinase resulted in fragments containing sequences similar to that of the preferred synthetic substrate peptide shown above. Therefore, under optimized in vitro conditions, the substrate recognition sequence for Pim-1 kinase is (Arg/Lys)3-X-Ser/Thr*-X', where X' is likely neither a basic nor a large hydrophobic residue.


Assuntos
Proteínas Serina-Treonina Quinases , Proteínas Tirosina Quinases/metabolismo , Proteínas Proto-Oncogênicas/genética , Proteínas Proto-Oncogênicas/metabolismo , Proto-Oncogenes , Sequência de Aminoácidos , Sequência de Bases , Cátions Bivalentes/farmacologia , Clonagem Molecular , Escherichia coli/genética , Glutationa Transferase/genética , Glutationa Transferase/metabolismo , Histonas/metabolismo , Humanos , Cinética , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Oligonucleotídeos Antissenso , Oligopeptídeos/metabolismo , Concentração Osmolar , Mapeamento de Peptídeos , Fosfopeptídeos/isolamento & purificação , Proto-Oncogene Mas , Proteínas Proto-Oncogênicas c-pim-1 , Proteínas Recombinantes de Fusão/metabolismo , Especificidade por Substrato
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