hydrochloric acid

盐酸
  • 文章类型: Journal Article
    降低RuO2/TiO2催化剂的成本仍然是催化HCl氧化的紧迫挑战之一。在目前的工作中,开发了一种低Ru负载量的Ce掺杂TiO2负载RuO2催化剂,在HCl催化氧化为Cl2方面表现出高活性。对Ce掺杂的TiO2载体及其负载型RuO2催化剂的一些广泛表征结果表明,Ce掺杂到TiO2中可以有效地改变TiO2的晶格参数,以改善活性RuO2物种在载体上的分散。这促进了表面Ru物质的产生,以暴露更多的活性位点,用于提高催化性能,即使在一些苛刻的反应条件下。该工作为氯气回收利用提供了一定的科学依据和技术支持。
    Reducing the cost of RuO2/TiO2 catalysts is still one of the urgent challenges in catalytic HCl oxidation. In the present work, a Ce-doped TiO2 supported RuO2 catalyst with a low Ru loading was developed, showing a high activity in the catalytic oxidation of HCl to Cl2. The results on some extensive characterizations of both Ce-doped TiO2 carriers and their supported RuO2 catalysts show that the doping of Ce into TiO2 can effectively change the lattice parameters of TiO2 to improve the dispersion of the active RuO2 species on the carrier, which facilitates the production of surface Ru species to expose more active sites for boosting the catalytic performance even under some harsh reaction conditions. This work provides some scientific basis and technical support for chlorine recycling.
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  • 文章类型: Journal Article
    经典微生物学的研究依赖于大细胞群体的平均行为,而不考虑克隆细菌群体可能通过随机转换机制分叉成表型不同的亚群。单核细胞增生李斯特菌暴露于亚致死胁迫可能会诱导共存的不同生理状态(即,亚致死性损伤或休眠)和存在可变的复苏能力。暴露于过乙酸(PAA;10-30ppm;持续3小时),在20°C下使用乙酸和盐酸(AA和HCl;pH3.0-2.5;持续5小时)在单核细胞增生李斯特菌中诱导不同的生理状态,斯科特A菌株。应力暴露后,监测单个细胞的集落生长,添加0.6%酵母提取物的胰蛋白酶大豆琼脂,使用延时显微镜,在37°C每5分钟采集图像并使用BaSCA框架进行分析。大多数获得的菌落生长曲线都拟合到Baranyi和Roberts的模型中,以估计滞后时间(λ)和最大比生长速率(μmax),除了暴露于AApH2.7和2.5后获得的那些适合Trilinear模型。遵循多元正态分布的λ和μmax数据用于使用蒙特卡罗模拟来预测生长变异性。用AA处理后的生长动力学(pH2.7和2.5;在20°C下持续5小时),PAA(30ppm;在20°C下3小时)表明,这些应力条件增加了右侧变异性分布的偏度,这意味着滞后时间的可变性增加,有利于更长的产出。暴露于AApH2.5和30ppmPAA导致每代两个不同的亚群,具有不同的生长动力学。这种转换机制可能已经发展成为单核细胞增生李斯特菌的生存策略,最大限度地提高生存的机会。微生物生长的模拟表明,当细胞从暴露于可能诱导损伤或休眠的亚致死胁迫(即PAA和酸性条件)中恢复时,生长动力学的异质性增加。
    Studies of classical microbiology rely on the average behaviour of large cell populations without considering that clonal bacterial populations may bifurcate into phenotypic distinct sub-populations by random switching mechanisms.Listeria monocytogenes exposure to sublethal stresses may induce different physiological states that co-exist (i.e., sublethal injury or dormancy) and present variable resuscitation capacity. Exposures to peracetic acid (PAA; 10-30 ppm; for 3 h), acetic acid and hydrochloric acid (AA and HCl; pH 3.0-2.5; for 5 h) at 20 °C were used to induce different physiological states in L. monocytogenes, Scott A strain. After stress exposure, colony growth of single cells was monitored, on Tryptic Soy Agar supplemented with 0.6 % Yeast Extract, using time-lapse microscopy, at 37 °C. Images were acquired every 5 min and were analyzed using BaSCA framework. Most of the obtained growth curves of the colonies were fitted to the model of Baranyi and Roberts for the estimation of lag time (λ) and maximum specific growth rate (μmax), except the ones obtained after exposure to AA pH 2.7 and 2.5 that were fitted to the Trilinear model. The data of λ and μmax that followed a multivariate normal distribution were used to predict growth variability using Monte Carlo simulations. Outgrowth kinetics after treatment with AA (pH 2.7 and 2.5; for 5 h at 20 °C), PAA (30 ppm; for 3 h at 20 °C) revealed that these stress conditions increase the skewness of the variability distributions to the right, meaning that the variability in lag times increases in favour of longer outgrowth. Exposures to AA pH 2.5 and 30 ppm PAA resulted in two distinct subpopulations per generation with different growth dynamics. This switching mechanism may have evolved as a survival strategy for L. monocytogenes cells, maximizing the chances of survival. Simulation of microbial growth showed that heterogeneity in growth dynamics is increased when cells are recovering from exposure to sublethal stresses (i.e. PAA and acidic conditions) that may induce injury or dormancy.
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  • 文章类型: Journal Article
    越来越多的研究强调了乳源细胞外囊泡(MEVs)在母婴整合中的重要性。以及它们作为新型药物递送系统和诊断生物标志物的应用。然而,常规超速离心(UC)通常导致MEV颗粒中酪蛋白胶束的共沉淀。在这项研究中,我们比较了不同原理的方法来筛选酪蛋白去除的最佳预处理,并发现盐酸(HA)等电沉淀可以最有效地去除猪奶中的酪蛋白。我们从不同方面进一步表征了UC和HA/UC分离的MEV群体,包括通过纳米粒子跟踪分析(NTA)和透射电子显微镜(TEM)的粒子方法,RNA和蛋白质含量,和纯度分析。重要的是,体外评估了MEVs的增殖和抗炎作用,与UC相比,通过HA/UC显示MEV在功能上的优越性。我们的结果表明,在超速离心前进行HA预处理可以有效去除酪蛋白和其他蛋白质复合物,通过HA/UC导致MEV具有更高的纯度和更显著的体外效果。这项研究为跨不同物种的MEV分离技术的进步和MEV的准确功能分析提供了有价值的见解。
    Increasing studies have highlighted the significance of milk-derived extracellular vesicles (MEVs) in mother-newborn integration, as well as their application as novel drug delivery systems and diagnostic biomarkers. However, conventional ultracentrifugation (UC) often results in the co-precipitation of casein micelles in MEV pellets. In this study, we compared methods with different principles to screen the optimal pretreatment in caseins removal, and found that isoelectric precipitation by hydrochloric acid (HA) could most effectively remove caseins in porcine milk. We further characterized MEV populations isolated by UC and HA/UC from diverse aspects, including particle methodology via nanoparticle tracking analysis (NTA) and transmission electron microscopy (TEM), RNA and protein contents, and purity analysis. Importantly, the proliferative and anti-inflammatory effects of MEVs were evaluated in vitro, showing the superiority of MEVs via HA/UC in functionality compared with UC. Our results suggest that HA pretreatment before ultracentrifugation could effectively remove caseins and other protein complexes, leading to MEVs via HA/UC with higher purity and more significant effects in vitro. This study provides valuable insights for the advancement of MEVs isolation techniques across different species and accurate function analysis of MEVs.
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  • 文章类型: Journal Article
    间质性膀胱炎/膀胱疼痛综合征(IC/BPS)是一种复杂的慢性疼痛疾病,具有难以捉摸的病因和非特异性症状。尽管已经建立了许多表型与人类疾病相似的动物模型,没有可用的治疗方案可以持续缓解临床症状。这种困境使我们质疑当前的动物模型是否足以代表IC/BPS。我们比较了四种常用的IC/BPS大鼠模型,以确定其不同的组织病理学和分子模式。雌性大鼠给予盐酸(HCL)单一处理,乙酸(AA),硫酸鱼精蛋白加脂多糖(PS+LPS),或环磷酰胺(CYP)诱导IC/BPS。膀胱切片染色用于组织病理学评估,使用下一代测序和基因集分析检查mRNA表达谱。HCL和AA组的肥大细胞计数明显高于PS+LPS,CYP,和对照组,但只有AA组表现出显著的胶原积聚。这些模型在基因本体论和京都百科全书的基因和基因组途径方面存在很大差异。我们的观察表明,这些大鼠模型都不能充分反映IC/BPS的复杂性。我们建议未来的研究同时应用和比较多个模型,以完全复制IC/BPS的复杂特征。
    Interstitial cystitis/bladder pain syndrome (IC/BPS) is a complex chronic pain disorder with an elusive etiology and nonspecific symptoms. Although numerous animal models with phenotypes similar to human disease have been established, no available regimen can consistently alleviate clinical symptoms. This dilemma led us to question whether current animal models adequately represent IC/BPS. We compared four commonly used IC/BPS rat models to determine their diverse histopathological and molecular patterns. Female rats were given single treatments with hydrochloric acid (HCL), acetic acid (AA), protamine sulfate plus lipopolysaccharide (PS + LPS), or cyclophosphamide (CYP) to induce IC/BPS. Bladder sections were stained for histopathologic evaluation, and mRNA expression profiles were examined using next-generation sequencing and gene set analyses. Mast cell counts were significantly higher in the HCL and AA groups than in the PS + LPS, CYP, and control groups, but only the AA group showed significant collagen accumulation. The models differed substantially in terms of their gene ontology and Kyoto encyclopedia of genes and genomes pathways. Our observations suggest that none of these rat models fully reflects the complexity of IC/BPS. We recommend that future studies apply and compare multiple models simultaneously to fully replicate the complicated features of IC/BPS.
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  • 文章类型: Journal Article
    现代等温滴定量热仪器提供了很高的精度,但对于可比的精度,他们需要化学校准。对于热因素,一种推荐的方法是HCl进入弱碱TRIS。在用一个VP-ITC和两个Nano-ITC研究这个反应时,我们遇到了一些问题,最重要的是滴定剂体积不足Δv≈0.3μL,我们将其归因于注射器尖端中HCl的扩散损失。扩散问题的数学处理支持了这种解释。效果是通过可变v协议发现的,因此,它应该被用来适当地允许它在任何类似接近完成的反应中。我们还发现,碳酸盐污染和弱碱水解产生的OH-的影响可能比以前认为的更显著。为了便于在数据的最小二乘拟合中进行适当的加权,我们从复制数据中估计了数据方差函数。所有这三种仪器的低信号精度均为σ≈1μJ;Nano-ITC的滴定剂体积不确定性比VP-ITC大2倍。Thefinalheatfactorsremainsuncertainbymorethanthe~1%precisionoftheinstrumentsandareundulatesensitivetotheHClconcentration.
    Modern isothermal titration calorimetry instruments give great precision, but for comparable accuracy they require chemical calibration. For the heat factor, one recommended process is HCl into the weak base TRIS. In studying this reaction with a VP-ITC and two Nano-ITCs, we have encountered some problems, most importantly a titrant volume shortfall Δv ≈ 0.3 μL, which we attribute to diffusive loss of HCl in the syringe tip. This interpretation is supported by a mathematical treatment of the diffusion problem. The effect was discovered through a variable-v protocol, which thus should be used to properly allow for it in any reaction that similarly approaches completion. We also find that the effects from carbonate contamination and from OH- from weak base hydrolysis can be more significant that previously thought. To facilitate proper weighting in the least-squares fitting of data, we have estimated data variance functions from replicate data. All three instruments have low-signal precision of σ ≈ 1 μJ; titrant volume uncertainty is a factor of ∼2 larger for the Nano-ITCs than for the VP-ITC. The final heat factors remain uncertain by more than the ∼1 % precision of the instruments and are unduly sensitive to the HCl concentration.
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  • 文章类型: Journal Article
    肽用于诊断,治疗学,作为抗菌剂。大多数肽是通过化学合成产生的,但是由于高滴度的优势,重组生产最近已成为一种有吸引力的选择,低毒废物和三级结构的正确折叠。生长抑素-28是一种调节内分泌系统的肽类激素,细胞增殖并抑制人体内多种次级激素的释放。它由28个氨基酸组成,有一个二硫键,这使得它成为整个下游纯化过程的最佳模型肽。我们使用CASPON™技术在大肠杆菌的周质中生产了该肽,一种亲和融合技术系统,能够实现重组蛋白的高可溶性表达,并与环状置换的人caspase-2切割融合标签。此外,产品的纯化是直接使用建立的平台过程。给出了两个不同的下游净化案例研究,从盐酸或聚乙烯亚胺作为萃取助剂开始。在从大肠杆菌的周质中释放亲和标记的生长抑素-28后,进行了几个纯化步骤,在最后的抛光步骤后递送纯肽溶液。通过反相高效液相色谱法以及质谱法监测该过程以确定产率和正确的二硫键形成。监测宿主细胞蛋白等杂质,每个下游单元后的DNA和内毒素证实了两种纯化途径的有效去除。
    Peptides are used for diagnostics, therapeutics, and as antimicrobial agents. Most peptides are produced by chemical synthesis, but recombinant production has recently become an attractive alternative due to the advantages of high titers, less toxic waste and correct folding of tertiary structure. Somatostatin-28 is a peptide hormone that regulates the endocrine system, cell proliferation and inhibits the release of numerous secondary hormones in human body. It is composed of 28 amino acids and has one disulfide bond, which makes it to an optimal model peptide for a whole downstream purification process. We produced the peptide in the periplasm of E. coli using the CASPON™ technology, an affinity fusion technology system that enables high soluble expression of recombinant proteins and cleaves the fusion tag with a circularly permuted human caspase-2. Furthermore, purification of the products is straight forward using an established platform process. Two different case studies for downstream purification are presented, starting with either hydrochloric acid or polyethyleneimine as an extraction aid. After release of affinity-tagged somatostatin-28 out of E. coli\'s periplasm, several purification steps were performed, delivering a pure peptide solution after the final polishing step. The process was monitored by reversed-phase high-performance liquid chromatography as well as mass spectrometry to determine the yield and correct disulfide bond formation. Monitoring of impurities like host cell proteins, DNA and endotoxins after each downstream unit confirmed effective removal for both purification pathways.
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  • 文章类型: Journal Article
    本研究的重点是一种环境方法,该方法基于使用Citrulluscolocynthis果实提取物中的生态友好型腐蚀抑制剂来增强酸性介质中碳钢(C-S)的耐腐蚀性,作为各种有机和非有机的替代品化学抑制剂。使用重量分析和电化学(动电位极化和交流阻抗)技术以及表面分析,在摩尔盐酸(1MHCl)介质中评估了瓜氨酸(CCE)果实甲醇提取物的抑制特性。CCE富含氨基酸,主要是瓜氨酸和β-(吡唑-1-基)-L-analine分子。根据减肥评估,结果表明,这种植物提取物作为一种有效的缓蚀剂,在303K金属暴露6小时后,在500ppm的CCE下达到了93.6%的保护水平。CCE作为混合型抑制剂发挥作用。此外,交流阻抗分析表明,将CCE掺入腐蚀性溶液中会导致负载能力降低,同时提高了接口的充电/放电功能。这表明在C-S表面上形成吸附层的可能性。此外,扫描电镜(SEM)观察,接触角测量,和傅立叶变换红外光谱(FTIR)分析支持在添加CCE之后在CS基材表面上形成保护膜。Langmuir和/或Temkin等温线可用于表征该有机抑制剂在C-S表面上的吸附。X射线光电子能谱(XPS)表明,CCE对1MHCl溶液中C-S腐蚀的抑制作用主要受化学吸附过程控制,并且抑制层由氧化铁/氢氧化物混合物组成,其中CCE分子被掺入。为了了解这些抑制剂分子的分子结构与防腐效果之间的关系,使用密度泛函理论(DFT)和分子动力学(MD)模拟进行了量子化学研究。
    The present study focuses on an environmental approach based on the use of an eco-friendly corrosion inhibitor from the Citrullus colocynthis fruit extract for enhancement corrosion resistance of carbon steel (C-S) in acid medium as an alternative to various organic and non-organic chemical inhibitors. The evaluation of the inhibition properties of the fruit methanolic extract of Citrullus colocynthis (CCE) were performed in molar hydrochloric acid (1 M HCl) medium using gravimetric and electrochemical (potentiodynamic polarization and AC impedance) techniques as well as surface analyses. CCE is rich in amino acids, mainly citrulline and β-(pyrazo-1-yl)-L-analine molecules. Based on the weight loss evaluation, the results demonstrated that this plant extract acts as an effective corrosion inhibitor and a protection level of 93.6% was attained at 500 ppm of CCE after 6 h of metal exposure at 303 K. According to polarization curves, CCE functions as a mixed-type inhibitor. In addition, AC impedance analyses have shown that the incorporation of CCE into the corrosive solution leads to a decrease in load capacity, while improving the charge/discharge function at the interface. This suggests the possibility of the formation of an adsorbed layer on the C-S surface. In addition, scanning electron microscope (SEM) observation, contact angle measurements, and Fourier-transform infrared spectroscopy (FTIR) analyses supported the development of a protective film over CS substrate surface afterwards addition of CCE. Langmuir and/or Temkin isotherms can be used to characterize the adsorption of this organic inhibitor on the C-S surface. X-ray photoelectron spectroscopy (XPS) has revealed that the inhibiting effect of CCE on the corrosion of C-S in 1 M HCl solution is mainly controlled by a chemisorption process and the inhibitive layer is composed of an iron oxide/hydroxide mixture where CCE molecules are incorporated. In order to understand the relationship between the molecular structure and anti-corrosion effectiveness of these inhibitor molecules, quantum chemical studies were carried out using density functional theory (DFT) and molecular dynamics (MD) simulation.
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  • 文章类型: Journal Article
    暴露于与食品加工相关的亚致死压力可能会诱导共存的细胞的异质混合物,包括健康,亚致命受伤,休眠和死亡的细胞。单细胞存活能力和休眠的异质性可能会阻碍食源性病原体的检测。在这项研究中,我们暴露了单核细胞增生李斯特菌ScottA菌株,过乙酸(PAA;20-40ppm)和酸性条件(盐酸(HCl)和乙酸(AA),调节至pH2.7-3.0,以评估两种不同培养基中代谢活性细胞的复苏能力和生长动力学。使用CFDA(代谢活性)和PI(死亡)染色通过流式细胞术评估细胞的损伤和存活但不可培养(VBNC)状态。强调的CFDA+PI-细胞在胰蛋白酶大豆(TS)琼脂或TS肉汤中分选,两者都补充了0.6%酵母提取物(TSAYE或TSBYE),评估可培养性。CFDA+PI分选的细胞的复苏能力(10个事件/孔)通过在TSBYE上的视觉检查和通过在TSBYE中的光密度测量来监测5天。在TSAYE和TSBYE上的林格氏溶液中分选单核细胞增生李斯特菌存活细胞(CFDA+PI-)显示在两种培养基(对照条件)中的100%回收率,而TSBYE中的平均滞后时间为9.6h。用20ppmPAA处理90和180min导致TSBYE中74.79%和85.82%的不可培养细胞,并将平均滞后时间增加到41.7h和43.8h,分别,与对照组相比(9.6h)。用30ppmPAA处理90分钟后检测到最长的平均滞后时间(79.5h),而在相同的条件下,CFDA+PI-细胞的分选在TSBYE和TSBYE上产生95.05%和93.94%的不可培养细胞,分别。在用40ppmPAA处理30分钟后,在TSAYE上检测到具有不可培养细胞的孔的最高百分比(96.17%)。用pH3.0的HCl处理60和240分钟后,在TSBYE中检测到VBNC细胞的分数,以及在暴露于AApH2.7后的TSAYE和TSBYE中。与60分钟相比,用AApH2.7处理150-300分钟增加了记录的滞后时间值的范围,从8.6小时到13.3小时,以及TSBYE中的平均滞后时间。比较两种类型的胁迫(氧化与酸)和两种生长系统(殖民地与浮游)的生长动力学模型表明,较低的起始浓度阻碍了对活的单核细胞增生李斯特菌细胞的检测。由于VBNC诱导或细胞异质性。
    Exposure to sublethal stresses related to food-processing may induce a heterogenous mixture of cells that co-exist, comprising healthy, sublethally injured, dormant and dead cells. Heterogeneity in survival capacity and dormancy of single cells may impede the detection of foodborne pathogens. In this study, we exposed Listeria monocytogenes Scott A strain, to peracetic acid (PAA; 20-40 ppm) and to acidic conditions (hydrochloric (HCl) and acetic (AA) acid, adjusted to pH 2.7-3.0, to evaluate the resuscitation capacity and outgrowth kinetics of metabolically active cells in two different media. Injury and the viable-but-non-culturable (VBNC) status of cells were assessed by flow cytometry using CFDA (metabolically active) and PI (dead) staining. Stressed CFDA+PI- cells were sorted on Tryptic Soy (TS) Agar or in TS broth, both supplemented with 0.6 % Yeast Extract (TSAYE or TSBYE), to evaluate culturability. Resuscitation capacity of CFDA+PI-sorted cells (10 events/well) was monitored by visual inspection on TSAYE and by optical density measurement in TSBYE for 5 days. Sorting of L. monocytogenes viable cells (CFDA+PI-) in Ringer\'s solution on TSAYE and TSBYE showed 100 % recovery in both media (control condition), while the mean lag time in TSBYE was 9.6 h. Treatment with 20 ppm PAA for 90 and 180 min resulted in 74.79 % and 85.82 % of non-culturable cells in TSBYE and increased the average lag time to 41.7 h and 43.8 h, respectively, compared to the control (9.6 h). The longest average lag time (79.5 h) was detected after treatment with 30 ppm PAA for 90 min, while at the same condition sorting of CFDA+PI- cells resulted in 95.05 % and 93.94 % non-culturable cells on TSAYE and TSBYE, respectively. The highest percentage of wells with non-culturable cells (96.17 %) was detected on TSAYE after treatment with 40 ppm PAA for 30 min. Fractions of VBNC cells were detected in TSBYE after treatment with HCl pH 3.0 for 60 and 240 min, and in TSAYE and TSBYE after exposure to AA pH 2.7. Treatment with AA pH 2.7 for 150-300 min increased the range of recorded lag time values compared to 60 min, from 8.6 h up to 13.3 h, as well as the mean lag times in TSBYE. Modelling of the outgrowth kinetics comparing the two types of stress (oxidative vs acid) and the two systems of growth (colonial vs planktonic) revealed that low starting concentrations hindered the detection of viable L. monocytogenes cells, either due to VBNC induction or cell heterogeneity.
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  • 文章类型: Journal Article
    诊断实验室中骨髓(BM)的固定和脱矿质方案尚未标准化。不同的方案如何影响组织形态学和DNA扩增尚不完全清楚。在这项研究中,在犬BM样品上测试了2种固定剂和3种脱矿质方法。将在死亡24小时内获得的20个重复胸骨样品在乙酸-锌-福尔马林(AZF)或10%中性缓冲的福尔马林(NBF)中固定过夜,并用甲酸脱矿质12小时。将另外53个样品固定在AZF中并用盐酸脱矿质1小时,甲酸12小时,或乙二胺四乙酸(EDTA)24小时。组织学切片由4名评估者评分为不足,边缘,不错,或优良的品质。此外,从用不同固定和脱矿质方法处理的切片中提取的DNA样品用3组引物扩增到T细胞受体γ和免疫球蛋白重链基因的保守区域。根据毛细管电泳图的回顾对扩增效率进行分级。固定在AZF或NBF中的切片的组织形态学评分没有显着差异。然而,基于EDTA的脱矿质比盐酸或甲酸的脱矿质产生更高的组织形态学评分,而甲酸的分数高于盐酸。用EDTA脱矿质在36个样品中的29个(81%)中产生了DNA扩增,而用任何一种酸进行的脱矿质仅在72个样品中的2个(3%)中产生扩增。虽然稍微耗时和劳动密集型,用EDTA进行组织脱矿质会导致优越的形态,并且对于使用本文所述的DNA提取方法进行聚合酶链反应(PCR)扩增至关重要。
    Fixation and demineralization protocols for bone marrow (BM) across diagnostic laboratories are not standardized. How different protocols affect histomorphology and DNA amplification is incompletely understood. In this study, 2 fixatives and 3 demineralization methods were tested on canine BM samples. Twenty replicate sternal samples obtained within 24 hours of death were fixed overnight in either acetic acid-zinc-formalin (AZF) or 10% neutral-buffered formalin (NBF) and demineralized with formic acid for 12 hours. Another 53 samples were fixed in AZF and demineralized with hydrochloric acid for 1-hour, formic acid for 12 hours, or ethylenediamine tetraacetic acid (EDTA) for 24 hours. Histologic sections were scored by 4 raters as of insufficient, marginal, good, or excellent quality. In addition, DNA samples extracted from sections treated with the different fixation and demineralization methods were amplified with 3 sets of primers to conserved regions of T cell receptor gamma and immunoglobulin heavy chain genes. Amplification efficiency was graded based on review of capillary electrophoretograms. There was no significant difference in the histomorphology scores of sections fixed in AZF or NBF. However, EDTA-based demineralization yielded higher histomorphology scores than demineralization with hydrochloric or formic acid, whereas formic acid resulted in higher scores than hydrochloric acid. Demineralization with EDTA yielded DNA amplification in 29 of 36 (81%) samples, whereas demineralization with either acid yielded amplification in only 2 of 72 (3%) samples. Although slightly more time-consuming and labor-intensive, tissue demineralization with EDTA results in superior morphology and is critical for polymerase chain reaction (PCR) amplification with the DNA extraction method described in this article.
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  • 文章类型: Journal Article
    氨基酸中氮的化合物特异性同位素分析(CSIA-AA,δ15NAA)在阐明食物链内的能量流和确定各种生物的营养位置方面越来越受欢迎。然而,对水解条件的影响缺乏研究,如HCl浓度和水解时间,生物群样品中的δ15NAA分析。在这项研究中,我们研究了两个HCl浓度(6M和12M)和四个水解时间(2小时,6h,12h,和24h)用于水解和衍生参考材料(金枪鱼)和小白鹭(n=4)的生物样品中的AA,夜鹭(n=4),sharpbelly(n=4)和藻类(n=1)使用正-新戊酰基-异丙基(NPIP)酯方法。Dowex阳离子交换树脂用于在衍生化之前纯化氨基酸。然后,我们使用气相色谱-燃烧-同位素比质谱(GC-C-IRMS)确定了δ15NAA值。结果表明,用不同HCl浓度或水解时间处理的样品之间的δ15NAA值没有显着差异(p>0.05)。特别是金枪鱼(12M)的δ15NGlx(范围:21.0-23.5‰)和δ15NPhe(范围:4.3-5.4‰)。根据2h时的δ15NAA计算的营养位置(TP)(小白鹭:2.9±0.1,夜鹭:2.8±0.1,尖胸:2.0±0.1和藻类:1.3±0.2)与24h时的营养位置(分别为3.1±0.1、2.8±0.1、2.2±0.1和1.1±0.1)一致,表明2小时的水解时间和6MHCl浓度是确定δ15NAA和估计TP的有效预处理条件。与目前使用的水解条件(24h,6米),建议的条件(2小时,6M)加速了δ15NAA测定,让它更快,更方便,更有效率。需要进一步研究以简化操作流程并降低时间成本,实现CSIA-AA的更有效应用。
    Compound-specific isotope analysis of nitrogen in amino acids (CSIA-AA, δ15NAA) has gained increasing popularity for elucidating energy flow within food chains and determining the trophic positions of various organisms. However, there is a lack of research on the impact of hydrolysis conditions, such as HCl concentration and hydrolysis time, on δ15NAA analysis in biota samples. In this study, we investigated two HCl concentrations (6 M and 12 M) and four hydrolysis times (2 h, 6 h, 12 h, and 24 h) for hydrolyzing and derivatizing AAs in reference materials (Tuna) and biological samples of little egret (n = 4), night heron (n = 4), sharpbelly (n = 4) and Algae (n = 1) using the n-pivaloyl-iso-propyl (NPIP) ester approach. A Dowex cation exchange resin was used to purify amino acids before derivatization. We then determined δ15NAA values using gas chromatography-combustion-isotope ratio mass spectrometry (GC-C-IRMS). The results revealed no significant differences (p > 0.05) in δ15NAA values among samples treated with different HCl concentrations or hydrolysis times, particularly for δ15NGlx (range: 21.0-23.5‰) and δ15NPhe (range: 4.3-5.4‰) in Tuna (12 M). Trophic positions (TPs) calculated based on δ15NAA at 2 h (little egret: 2.9 ± 0.1, night heron: 2.8 ± 0.1, sharpbelly: 2.0 ± 0.1 and Algae: 1.3 ± 0.2) were consistent with those at 24 h (3.1 ± 0.1, 2.8 ± 0.1, 2.2 ± 0.1 and 1.1 ± 0.1, respectively), suggesting that a 2-h hydrolysis time and a 6 M HCl concentration are efficient pretreatment conditions for determining δ15NAA and estimating TP. Compared to the currently used hydrolysis conditions (24 h, 6 M), the proposed conditions (2 h, 6 M) accelerated the δ15NAA assay, making it faster, more convenient, and more efficient. Further research is needed to simplify the operational processes and reduce the time costs, enabling more efficient applications of CSIA-AA.
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