Genomic DNA

基因组 DNA
  • 文章类型: Journal Article
    Recent studies support the hypothesis that microbes can seed some Alzheimer\'s disease (AD) cases, leading to inflammation and overproduction of amyloid peptides. Porphyromonas gingivalis (Pg) is a keystone pathogen of chronic periodontitis and has been identified as risk factor for the development and progression of AD. The present preliminary study aimed to quantify Pg abundance in neurodegenerative disease (ND) patients compared with neurologic patients without neurodegenerative disorders (no-ND) and healthy controls (HC) to determine possible association between Pg abundance and neurodegenerative process. Pg was quantified on DNA extracted from the oral samples of 49 patients and 29 HC by quantitative polymerase chain reaction (qPCR). Anti-Pg antibodies were also detected on patient serum samples by enzyme-linked immunosorbent assays (ELISA). The Pg abundance in the oral cavity was significantly different among groups (p = 0.004). It was higher in ND than no-ND (p = 0.010) and HC (p = 0.008). The Pg abundance was correlated with the antibodies (p = 0.001) with different slopes between ND and no-ND (p = 0.037). Pg abundance was not correlated with oral indices and comorbidities. These results extend our understanding of the association between oral pathogens and AD to other neurodegenerative processes, confirming the hypothesis that oral pathogens can induce an antibody systemic response, influencing the progression of the disease.
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  • 文章类型: Journal Article
    本研究旨在评估埃及哮喘儿童的血清维生素D和免疫球蛋白E(IgE)水平,并找出维生素D受体(VDR)多态性与支气管哮喘发展的可能关联。
    这项研究包括了100名埃及儿童,50名哮喘儿童与50岁相当,性别,和体重指数匹配,使用全球哮喘倡议对哮喘儿童进行非相关健康对照(HCs)临床评估.肺功能检查(1秒用力呼气量[FEV1],强制肺活量[FVC],进行FEV1/FVC比率)。进行血清维生素D3和总IgE的酶联免疫吸附测定。VDR-单核苷酸多态性(SNP)(ApaI,TaqI,和BsmI)检测已通过限制性片段长度多态性技术使用聚合酶链反应进行。使用SPSS20.0版进行数据分析。所研究的SNP遵循HardyWeinberg方程。
    哮喘儿童的平均血清25(OH)D3水平(13.46ng/mL±10.50SD)明显低于HCs(37.53ng/mL±13。0.40SD),P<0.05。在72%的病例中检测到维生素D缺乏,其在哮喘控制方面的水平没有显着差异。哮喘儿童的IgE水平(99.83ku/L±233.81SD)明显高于HCs(7.52ku/L±3.32SD),P<0.05。哮喘儿童更常见于TaqIt等位基因(比值比[OR],2.25;95%置信区间[CI],1.28-3.96;P<.05)和BsmIb等位基因(OR,1.83;95%CI,1.05-3.21;P<0.05)。ApaIa等位基因在患者与对照组之间没有显着差异(P>0.05)。TT+Tt和Bb+bb基因型在病例中明显高于对照组,所有P<0.05。
    TaqI和BsmI与埃及儿童支气管哮喘发展的风险相关。哮喘儿童中经常发生高IgE和低维生素D状态。
    The current study aimed to assess the serum levels of vitamin D and immunoglobulin E (IgE) among asthmatic Egyptian children and to find out the possible associations of vitamin D receptor (VDR) polymorphisms with bronchial asthma development.
    The study included 100 Egyptian children, 50 asthmatic children who were comparable with 50 age, sex, and body mass index-matched, unrelated healthy controls (HCs) clinical assessments of asthmatic children were done using global initiative of asthma. Pulmonary function tests (forced expiratory volume in 1 second [FEV1], forced vital capacity [FVC], FEV1/FVC ratio) were performed. Enzyme-linked immunosorbent assays of serum vitamin D3 and total IgE were done. VDR-single nucleotide polymorphisms (SNPs) (ApaI, TaqI, and BsmI) detection has performed using polymerase chain reaction through restriction fragment length polymorphism technique. Data analysis was performed using SPSS version 20.0. The studied SNPs were followed the Hardy Weinberg equation.
    The mean serum level of 25(OH) D3 was significantly lower among asthmatic children (13.46 ng/mL ± 10.50 SD) in comparison to HCs (37.53 ng/mL ± 13. 0.40 SD), P < .05. Vitamin D deficiency was detected in 72% of cases with no significant difference in its level regarding asthma control. There was significantly higher IgE level among asthmatic children (99.83 ku/L ± 233.81 SD) versus HCs (7.52 ku/L ± 3.32 SD), P < .05. Asthmatic children were presented more commonly with TaqI t allele (odds ratio [OR], 2.25; 95% confidence interval [CI], 1.28-3.96; P < .05) and BsmI b allele (OR, 1.83; 95% CI, 1.05-3.21; P < .05). ApaI a allele was not significantly different among patients versus controls (P > .05). TT + Tt and Bb + bb genotypes were significantly higher among cases versus the controls, P < .05 for all.
    TaqI and BsmI were associated with risk of bronchial asthma development among Egyptian children. High IgE and Low vitamin D status were frequently occurring among asthmatic children.
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  • 文章类型: Journal Article
    In this work, surface-enhanced Raman spectra of ten genomic DNAs extracted from leaf tissues of different grapevine (Vitis vinifera L.) varieties, respectively, are analyzed in the wavenumber range 300-1800 cm-1. Furthermore, structural changes induced in grapevine genomic nucleic acids upon femtosecond (170 fs) infrared (IR) laser pulse irradiation (λ = 1100 nm) are discussed in detail for seven genomic DNAs, respectively. Surface-enhanced Raman spectroscopy (SERS) signatures, vibrational band assignments and structural characterization of genomic DNAs are reported for each case. As a general observation, the wavenumber range between 1500 and 1660 cm-1 of the spectra seems to be modified upon laser treatment. This finding could reflect changes in the base-stacking interactions in DNA. Spectral shifts are mainly attributed to purines (dA, dG) and deoxyribose. Pyrimidine residues seem to be less affected by IR femtosecond laser pulse irradiation. Furthermore, changes in the conformational properties of nucleic acid segments are observed after laser treatment. We have found that DNA isolated from Feteasca Neagra grapevine leaf tissues is the most structurally-responsive system to the femtosecond IR laser irradiation process. In addition, using unbiased computational resources by means of principal component analysis (PCA), eight different grapevine varieties were discriminated.
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  • 文章类型: Journal Article
    OBJECTIVE: The aim of this clinical investigation was to identify and quantify the microbial species adhering to toothbrush bristles after controlled brushing and storage in different antimicrobial agents.
    METHODS: Sixteen healthy participants were enrolled in this study and randomly submitted to 4 interventions in a cross-over design: brushing and toothbrush storage in (I) Periogard/(II) Periobio (Chlorhexidine gluconate 0.12%), (III) Cepacol (cetylpyridinium chloride 0.05%) and (IV) distilled water (positive control). Thirty-eight bacterial species including putative pathogens and 5 Candida spp. were assessed by Checkerboard DNA-DNA hybridization.
    RESULTS: The results of the study have shown a striking reduction of the total microbial counts, including bacteria and Candida spp., on the toothbrush bristles after storage in cetylpyridinium chloride 0.05% (p < 0.0001). Chlorhexidine gluconate 0.12% showed no differences on the total bacterial count when compared to distilled water (p > 0.05). Cetylpyridinium chloride solution also presented the lowest genome counts and frequency of detection for individual target species; distilled water showed the highest individual genome counts (p < 0.05). Potential pathogenic species were recorded in moderate to high levels for chlorhexidine gluconate and distilled water.
    CONCLUSIONS: Cetylpyridinium chloride 0.05% was the most effective storage solution in the reduction of total and individual microbial counts, including pathogenic species.
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  • 文章类型: Journal Article
    细菌病原体引起越来越多的食品安全和生物恐怖主义问题。目前利用灵敏的纳米技术和生物传感器的DNA检测方法已显示出优异的检测性能,但需要昂贵且耗时的聚合酶链反应(PCR)来扩增DNA靶标;因此,更快,更经济的方法仍然是必不可少的。在这个概念验证研究中,我们研究了金纳米颗粒-DNA(AuNP-DNA)生物传感器检测非PCR扩增的基因组肠道沙门氏菌的能力(S.肠炎)DNA,来自纯或混合细菌培养物和加标液体基质。将非PCR扩增的DNA杂交成夹心样结构(磁性纳米颗粒/DNA/AuNP),并通过使用差分脉冲伏安法检测金伏安峰来分析。我们的初步数据表明,可以从细菌培养物和加标液体基质中检测到浓度低至100ng/mL的非PCR扩增的基因组DNA,与报告的PCR扩增检测水平相似。这些发现还表明,AuNP-DNA生物传感器是细菌病原体可行检测方法的第一步,特别是,对于资源有限的设置,例如基于现场或经济有限的条件。未来的工作将集中在进一步优化DNA提取方法和AuNP-生物传感器,与PCR扩增的DNA检测策略相比,可以在较低的食物基质DNA靶浓度下提高灵敏度。
    Bacterial pathogens pose an increasing food safety and bioterrorism concern. Current DNA detection methods utilizing sensitive nanotechnology and biosensors have shown excellent detection, but require expensive and time-consuming polymerase chain reaction (PCR) to amplify DNA targets; thus, a faster, more economical method is still essential. In this proof-of-concept study, we investigated the ability of a gold nanoparticle-DNA (AuNP-DNA) biosensor to detect non-PCR amplified genomic Salmonella enterica serovar Enteritidis (S. enteritidis) DNA, from pure or mixed bacterial culture and spiked liquid matrices. Non-PCR amplified DNA was hybridized into sandwich-like structures (magnetic nanoparticles/DNA/AuNPs) and analyzed through detection of gold voltammetric peaks using differential pulse voltammetry. Our preliminary data indicate that non-PCR amplified genomic DNA can be detected at a concentration as low as 100 ng/mL from bacterial cultures and spiked liquid matrices, similar to reported PCR amplified detection levels. These findings also suggest that AuNP-DNA biosensors are a first step towards a viable detection method of bacterial pathogens, in particular, for resource-limited settings, such as field-based or economically limited conditions. Future efforts will focus on further optimization of the DNA extraction method and AuNP-biosensors, to increase sensitivity at lower DNA target concentrations from food matrices comparable to PCR amplified DNA detection strategies.
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