RESUMO
The synergistic effects on the 0.18 µm PPD CISs induced by neutron displacement damage and gamma ionization damage are investigated. The typical characterizations of the CISs induced by the neutron displacement damage and gamma ionization damage are presented separately. The CISs are irradiated by reactor neutron beams up to 1 × 1011 n/cm2 (1 MeV neutron equivalent fluence) and 60Co γ-rays up to the total ionizing dose level of 200 krad(Si) with different sequential order. The experimental results show that the mean dark signal increase in the CISs induced by reactor neutron radiation has not been influenced by previous 60Co γ-ray radiation. However, the mean dark signal increase in the CISs induced by 60Co γ-ray radiation has been remarkably influenced by previous reactor neutron radiation. The synergistic effects on the PPD CISs are discussed by combining the experimental results and the TCAD simulation results of radiation damage.
RESUMO
Previously, we studied an AAVS1 site-specific non-viral integration system with a Rep-donor plasmid and a plasmid containing adeno-associated virus integration element. Our earlier study focused on the plasmid vector itself, but the cellular response to the system was still unknown. SP100 is a member of the promyelocytic leukemia nuclear bodies. It is involved in many cellular processes such as transcriptional regulation and the cellular intrinsic immune response against viral infection. In this study, we revealed that SP100 inhibited the Rep-dependent nonviral integration. Conversely, transient expression of Rep78 increased the degradation of SP100. This degradation was inhibited by treatment with MG132, an inhibitor of the ubiquitin proteasome. SP100 and Rep78 are both located in the nucleolus, which provides the spatial possibility for their interaction. Rep78 was coimmunoprecipitated with the enhanced green fluorescent protein (EGFP)-SP100 fusion protein but not EGFP, which verified the interaction between Rep78 and SP100. These results have enriched our knowledge about the cellular protein SP100 and Rep-dependent nonviral integration. It may lead to an improvement in the application of Rep-related transgene integration method and in the selection of target cells.
Assuntos
Antígenos Nucleares/metabolismo , Autoantígenos/metabolismo , Dependovirus/fisiologia , Integração Viral , Linhagem Celular , Proteínas de Ligação a DNA/metabolismo , Dependovirus/genética , Humanos , Plasmídeos , Ligação Proteica , Proteínas Virais/metabolismoRESUMO
BACKGROUND: Pyrexia is often associated with unfavorable stroke outcomes. However, limited information is available on the relationship between the causes of poststroke hyperthermia and stroke prognosis, especially for mild-to-moderate neurogenic pyrexia in acute cerebral infarction. AIMS: To compare the differences in the clinical features and characteristics of pyrexia as well as its prognosis among acute cerebral infarction patients with mild-to-moderate neurogenic pyrexia, with infectious pyrexia, and without pyrexia. The focus was on mild-to-moderate neurogenic pyrexia. METHODS: A total of 709 patients with acute cerebral infarction were prospectively recruited and their clinical data were analyzed. RESULTS: No significant difference was detected in age, gender, history of smoking, hypertension, or diabetes among the 3 groups (p > 0.05). Patients with mild-to-moderate neurogenic pyrexia and those with infectious pyrexia had higher baseline National Institutes of Health Stroke Scale (NIHSS) scores (15.1 ± 6.7, p = 0.003; 14.3 ± 8.1, p = 0.002, respectively), lower 3-month Barthel index (BI) values (64.2 ± 40.7, p < 0.001; 61.9 ± 49.3, p < 0.001, respectively) and higher 3-month mortality rates (13%, p = 0.026; 16%, p < 0.001, respectively) than patients without pyrexia (NIHSS score 11.4 ± 7.9; BI 82.6 ± 39.8, and mortality rate 6%, respectively). No difference existed in these parameters between the 2 pyrexia groups (p > 0.05), but mild-to-moderate neurogenic pyrexia had an earlier onset and a shorter duration than infectious pyrexia (p < 0.001). CONCLUSIONS: Acute cerebral infarction patients with mild-to-moderate neurogenic pyrexia had a similar prognosis compared to those with infectious pyrexia. Mild-to-moderate neurogenic pyrexia is possibly associated with stroke severity.
Assuntos
Infarto Cerebral/complicações , Febre/etiologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Infarto Cerebral/mortalidade , Feminino , Febre/mortalidade , Humanos , Masculino , Pessoa de Meia-Idade , PrognósticoRESUMO
The adeno-associated virus (AAV) p5 promoter controls expression of Rep68 and Rep78, which are responsible for specific integration of the viral genome into the AAVS1 site of the human genome. The p5 promoter contains a Rep-binding element (RBE) sequence that acts as a substrate of the Rep proteins for both site-specific integration of p5 itself and transcriptional suppression of the p5 promoter. To differentiate these two Rep-mediated functions, we dissected the p5 core structure TATA/RBE/YY1+1 through a series of mutations. Mutations in the TATA box or YY1+1 region of p5IEE significantly reduced Rep-mediated site-specific integration (RMSSI) and p5 promoter transcriptional activity, but only the TATA box is involved in Rep-mediated transcriptional suppression (RMTS). Point mutations at nucleotides 266, 267, 268, 270, and 273 of the GAGTGAGC motif in p5 RBE significantly reduced RMSSI efficiency. However, only p5G270T lost the affinity of Rep binding and had significant reduction of RMTS. It appears that RMTS is determined by the affinity of p5RBE for Rep whereas RMSSI requires more stringent conditions. Thus, RMTS and RMSSI can be differentiated by point mutations in the p5 promoter, which is useful in gene therapy in a helper vector to drive Rep expression, as the mutant promoters seldom integrate themselves but remain the RMTS feature for reduced cytotoxicity caused by a high level of Rep protein.
Assuntos
Sequências Reguladoras de Ácido Nucleico/genética , Sequência de Bases , Dependovirus/genética , Dependovirus/metabolismo , Expressão Gênica , Genoma Viral , Humanos , Ligação Proteica/genética , TATA BoxRESUMO
A new method using gel permeation chromatography (GPC) cleanup followed by ultra-performance liquid chromatography combined with tandem mass spectrometry (UPLC-MS-MS) has been established for simultaneous determination of 18 carbamate pesticides in nuts (chestnut and pine nut). Recoveries obtained by fortifying nut (spiking at 0.02 mg/kg) range from 70.21% to 89.56%. The proposed method features good sensitivity. Its limits of quantification are low enough to allow pesticide residues to be determined at concentrations below the maximum residue levels legally accepted. The precision, expressed as relative standard deviation, ranges from 2.26% to 4.07%.