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Surface tensometry studies on formulations of surfactants with preservatives as a tool for antimicrobial drug protection characterization 含有防腐剂的表面活性剂配方的表面张力法研究作为抗菌药物保护特性的工具
Pub Date : 2012-11-29 DOI: 10.4236/JBPC.2012.34040
S. Pogorzelski, D. Watrobska-Swietlikowska, M. Sznitowska
The aim of the work was to quantify possible interactions between surfactants and preservatives, comparing surface properties, in model pharmaceutical formulations. Surface parameters of 2-component surfactant-preservative aquous mixtures were determined with a Wilhelmy plate technique, for the so-called principal surfactants (polysorbate 80, egg lecithin, phosphatidylcholine) and preservatives, which were methylparaben and benzalkonium chloride (BA-C). A generalized surface tension vs. surfactant concentration plot signatures, in the presence of preservative at a fixed amount, allowed: the critical micellar concentration (cmc) shift, additive molecules partition from the surface to the bulk, mixed micelles formation concentration, and additive surface removal concentration to be determined in reference to surface activity of the added substance. Methylparaben is a compound of lower (in comparison to BAC) surface activity, lower partitioning coefficient possessing lower energy and concentration of its removal from the surface, that makes it play effectively an antimicrobial protection role in the bulk of pharmaceutical products, as already shown by chemical tests.
这项工作的目的是量化表面活性剂和防腐剂之间可能的相互作用,比较表面性质,在模型药物配方。采用Wilhelmy平板技术测定了两组分表面活性剂-防腐剂水混合物的表面参数,其中主要表面活性剂为聚山梨酸酯80、卵磷脂、磷脂酰胆碱,防腐剂为对羟基苯甲酸甲酯和苯并氯铵(BA-C)。在一定量防腐剂存在的情况下,一个广义的表面张力与表面活性剂浓度图特征允许:临界胶束浓度(cmc)的移动,添加剂分子从表面到体的分配,混合胶束形成浓度,以及添加剂表面去除浓度,这些都是根据所添加物质的表面活性来确定的。对羟基苯甲酸甲酯是一种表面活性较低(与BAC相比)的化合物,其分配系数较低,其从表面去除的能量和浓度较低,这使得它在大部分药品中有效地发挥抗菌保护作用,化学试验已经表明。
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引用次数: 15
Impact of petroleum refining activities on nitrate and nitrite content of edible vegetables and on their in vivo kinetics in albino rats 石油炼制活动对食用蔬菜中硝酸盐和亚硝酸盐含量及其对白化大鼠体内动力学的影响
Pub Date : 2012-11-29 DOI: 10.4236/JBPC.2012.34032
G. Otti, P. N. Okafor
The influence of pollution from petroleum refining activities on the levels of nitrates and nitrites in five edible vegetable species was investigated. Besides, the kinetics of nitrite and nitrate was studied in vivo using albino rats with focus on the possible influence of concentration difference on kinetics and implications to toxicity. Leaf samples of the five vegetable species were collected randomly from various locations within Eleme, a host community of Port Harcourt Refinery Company and the Indorama Petrochemical Company. Also, samples were collected from Umuahia, which served as pollution-free control. The leaf samples were analyzed for their nitrite and nitrate contents. Nitrite was determined spectrophotometrically while nitrate was determined after cadmium column reduction. Results showed that samples from Eleme had higher mean nitrate (349.20 mg/100g dry leaf mass; P 0.05) as compared to the same samples from Umuahia. Solutions of nitrate and nitrite, equivalent in concentration to mean nitrate and nitrite content of the vegetable samples from the two locations were administered enterally to four groups of albino rats. Analysis of their blood levels were monitored five times at 30 minutes intervals following administration. Rates of change of blood nitrites and nitrates were found to be fairly constant in absorption as well as in the elimination phase. Their peak blood concentrations varied proportionately with their concentrations in administered solutions. However, peak blood nitrate was attained later in group of animals receiving higher amount of nitrate solution. Refining activities may pre-dispose people living within Eleme community to health hazards through contamination of edible vegetables.
研究了石油炼制污染对5种食用蔬菜中硝酸盐和亚硝酸盐含量的影响。此外,我们还在白化大鼠体内研究了亚硝酸盐和硝酸盐的动力学,重点研究了浓度差异对动力学的可能影响及其对毒性的影响。在哈科特港炼油公司和印多拉马石油化工公司的主社区Eleme的不同地点随机收集了五种蔬菜的叶子样本。同时,从Umuahia采集样本,作为无公害对照。对叶片样品进行亚硝酸盐和硝酸盐含量分析。亚硝酸盐采用分光光度法测定,硝酸盐采用镉柱还原法测定。结果表明,Eleme样品的平均硝酸盐含量较高,为349.20 mg/100g干叶质量;P 0.05)。四组白化大鼠肠内注射硝酸盐和亚硝酸盐溶液,其浓度相当于两个地点蔬菜样品中硝酸盐和亚硝酸盐的平均含量。在给药后每隔30分钟监测5次血液水平分析。血液中亚硝酸盐和硝酸盐的变化率在吸收和消除阶段是相当恒定的。其血药浓度峰值随溶液浓度成比例变化。然而,高剂量硝酸盐溶液组血硝酸盐峰值出现较晚。精炼活动可能使生活在Eleme社区的人们预先受到可食用蔬菜污染的健康危害。
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引用次数: 3
The influence of chromium ions on the growth of the calcium hydroxyapatite crystal 铬离子对羟基磷灰石钙晶体生长的影响
Pub Date : 2012-11-29 DOI: 10.4236/JBPC.2012.34034
S. Sallam, Khairi M. Tohami, A. Sallam, L. I. A. Salem, F. A. Mohamed
Hydroxyapatite (Ca10(PO4)6(OH)2) is widely used as bio-ceramic materials and as adsorbents for separation of bio-molecules. These materials have also been used as adsorbents for heavy metals, supports and as catalysts in oxidation and dehydrogenation reactions. The catalytic performance of these materials depend on the lattice substitution of Ca sites in Hydroxyapatite structure by varied cations as Na, Mg, Sr and Mn, which result in changes in various structural pro perties as crystallinity and morphology. Pure calcium hydroxyapatite (S1) and Cr loaded hydroxyl apatite (S2, S3, S4 and S5) of different chromium concentrations have been prepared by wet precipitated method. An in-vitro examination is essential to investigate the mechanism of the deficient HA and tissue interface reaction by preparing SBF (Simulated Body Fluid) through the elemental and chemical analysis of Ca, P and Cr. FTIR used to analyze the samples after incubation in SBF for 24 day. PH of the samples also was measured at the first period of immersion time. At high loading of chromium ions, the formation of carbonate apatite decrease. The concentrations of the chromium in the Cr_HA crystal during the soaking in SBF are very safe dose for human.
羟基磷灰石(Ca10(PO4)6(OH)2)被广泛用作生物陶瓷材料和生物分子分离的吸附剂。这些材料也被用作重金属的吸附剂、载体和氧化和脱氢反应的催化剂。这些材料的催化性能取决于不同阳离子如Na、Mg、Sr和Mn对羟基磷灰石结构中Ca位点的晶格取代,从而导致结晶度和形貌等各种结构性能的变化。采用湿沉淀法制备了不同铬浓度的纯钙羟基磷灰石(S1)和载Cr羟基磷灰石(S2、S3、S4和S5)。通过Ca, P和Cr的元素和化学分析,制备SBF(模拟体液)来研究HA缺乏和组织界面反应的机制是必要的。在SBF中孵育24天后,使用FTIR分析样品。在第一次浸泡时,测定了样品的PH值。在高铬离子负荷下,碳酸盐磷灰石的形成减少。在SBF浸泡过程中,结晶中铬的浓度是人体非常安全的剂量。
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引用次数: 5
Tityus serrulatus venom and its toxins Ts1 and Ts5 increase cytosolic Ca2+ concentration in isolated vascular smooth muscle cells 浆鱼毒液及其毒素Ts1和Ts5可增加离体血管平滑肌细胞胞浆内Ca2+浓度
Pub Date : 2012-11-29 DOI: 10.4236/JBPC.2012.34035
M. A. Neto, F. Vasconcelos, L. Bendhack, E. Arantes
Voltage-gated Na+ channel (Nav channel) scorpion toxins are classified as α- and β-neurotoxins. Ts5 (α-neurotoxin) and Ts1 (β-neurotoxin) from Tityus serrulatus venom (TsV) interact with Nav channels, increasing Na+ influx and activating voltage-dependent Ca2+ channels. This study aimed to investigate the effect of TsV, Ts1 and Ts5 on the cytosolic Ca2+ concentration ([Ca2+]C) in rat aortic smooth muscle cells. Toxins were isolated by ion exchange chromatography (Ts1) followed by RP-HPLC (Ts5). The rat aortic smooth muscle cells were isolated in Hanks buffer pH 7.4 and loaded with 5 μmol/L of Fura-2AM (45 minutes at 37℃), in order to measure [Ca2+]C by fluorescence of Fura-2/AM (ratio 340/380 nm). The fluorescence was measured in one single cell (excitation: 340 and 380 nm; emission: 510 nm). TsV (100 and 500 mg/mL) and its toxins Ts1 and Ts5 (50 and 100 mg/mL each) led to a concentration-dependent increase in [Ca2+]C. Tetrodotoxin (1 mmol/L), a Nav channel blocker, and verapamil (1 mmol/L), a voltage-operated Ca2+ channel blocker, inhibited the increase in [Ca2+]C induced by TsV (500 mg/mL). In conclusion, TsV and its toxins induce a concentration-dependent increase in [Ca2+]C that probably occurs through interaction with Nav channels, thus inducing depolarization and consequent Ca2+ influx. This assumption is based on the fact that this effect is inhibited by tetrodotoxin and verapamil, showing a direct action of TsV toxins on aorta smooth muscle cells.
电压门控Na+通道(Nav通道)蝎毒素分为α-和β-神经毒素。毒TsV中的Ts5 (α-神经毒素)和Ts1 (β-神经毒素)与Nav通道相互作用,增加Na+内流,激活电压依赖性Ca2+通道。本研究旨在探讨TsV、Ts1和Ts5对大鼠主动脉平滑肌细胞胞浆Ca2+浓度([Ca2+]C)的影响。采用离子交换色谱(Ts1)和反相高效液相色谱(Ts5)分离毒素。取大鼠主动脉平滑肌细胞,在pH 7.4的Hanks缓冲液中分离,负载5 μmol/L Fura-2AM(37℃,45分钟),用Fura-2/AM (340/380 nm)荧光测定[Ca2+]C。在单个细胞中测量荧光(激发:340和380 nm;发射光谱:510纳米)。TsV(100和500 mg/mL)及其毒素Ts1和Ts5(各50和100 mg/mL)导致[Ca2+]C的浓度依赖性增加。Nav通道阻滞剂河豚毒素(1 mmol/L)和电压操作Ca2+通道阻滞剂维拉帕米(1 mmol/L)抑制TsV (500 mg/mL)诱导的[Ca2+]C升高。总之,TsV及其毒素诱导[Ca2+]C的浓度依赖性增加,这可能通过与Nav通道的相互作用发生,从而诱导去极化和随后的Ca2+内流。这一假设是基于这一作用被河豚毒素和维拉帕米抑制的事实,表明TsV毒素对主动脉平滑肌细胞有直接作用。
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引用次数: 3
Recent enhancement of the immunity in AIDS and other immunocompromised patients by hyperforin an antibiotic from Hypericum perforatum L. (in vitro model) part I 从贯叶连翘(Hypericum perforatum L.)中提取的抗生素贯叶连翘素(Hypericum perforatum L.)对艾滋病和其他免疫功能低下患者的近期免疫增强作用(体外模型)
Pub Date : 2012-11-29 DOI: 10.4236/JBPC.2012.34037
I. Brondz, Anton Brondz
Today, Hypericum perforatum L. is probably one of the best-characterized medicinal plants, and hyperforin is its best-characterized constituent. Extracts from H. perforatum are widely used as antidepressants; however, less attention has been given to other properties of hyperforin, such as antitumor, fungicidal, antiviral and antibacterial action, or its possible use as a substance with immunomodulation properties. The present study summarizes results that describe the influence of hyperforin as an immunomodulation agent on phagocytosis and the breakdown of Escherichia coli by human polymorphonuclear neutrophils (PMNs). Hyperforin at 1 - 100 μg/mL concentrations was found to have a major influence on phagocytosis and the breakdown of E. coli by PMNs in vitro. A 100 μg/mL solution of hyperforin increased the uptake of non-opsonized E. coli almost 50-fold, and the uptake of IgG-opsonized E. coli more than threefold; on the other hand, the uptake of serum-opsonized bacteria was reduced to approximately 60% of that of the control. Hyperforin seems to bind to both PMNs and E. coli and acts like an opsonin. The elimination of remnants of IgG-opsonized E. coli from the PMNs was stimulated by hyperforin, while the elimination of remnants from non-op-so nized and serum-opsonized material was unaffected by the drug. Hyperforin exhibited clear immunomodulation ability as a phagocytosisstimulating agent. Hyperforin is probably inactive against human immunodeficiency virus (HIV) and most Gram-negative bacteria. However, it can protect acquired immunodeficiency syndrome (AIDS) patients and other immunocompromised patients by its antibacterial activity against Gram-positive bacteria and by enhancement of phagocytosis of Gram-positive and Gram-negative bacteria; some Gram-negative bacteria, such as Neisseria, are sensitive to hyperforin. Hyperforin has the ability to penetrate the blood-brain barrier (BBB) and blood-testis barrier (BTB) and is a valuable antibacterial agent against meningitis and gonorrhea. These properties of hyperforin are important for an antibiotic with immunomodulation activity in the struggle against the growing mortality in AIDS patients as a result of opportunistic bacteria, as recently shown by Bekondi et al. (2006, Int. J. Infect. Dis. 10, 387-395). It could also help to combat primary and opportunistic pathogens associated with meningitis in adults' relation to HIV serostatus.
贯叶连翘(Hypericum perforatum L.)可能是当今最具特征的药用植物之一,而贯叶连翘素是其最具特征的成分。贯叶连翘提取物被广泛用作抗抑郁药;然而,对hyperhyperin的其他特性,如抗肿瘤、杀真菌、抗病毒和抗菌作用,或其作为一种具有免疫调节特性的物质的可能性的关注较少。本研究总结了作为一种免疫调节剂对人多形核中性粒细胞(PMNs)吞噬和分解大肠杆菌的影响。研究发现,1 ~ 100 μg/mL浓度的hyperperin对PMNs体外吞噬和分解大肠杆菌有重要影响。100 μg/mL的hyperperforin溶液使非调理大肠杆菌的摄取增加了近50倍,使igg调理大肠杆菌的摄取增加了3倍以上;另一方面,血清调理菌的摄取减少到对照组的约60%。hyperperin似乎与pmn和大肠杆菌结合,并像调理素一样起作用。hyperperin刺激PMNs中IgG-opsonized的残余大肠杆菌的消除,而非oponized和血清opsonized的残余物质的消除不受药物的影响。hyperperin作为一种促吞噬剂表现出明显的免疫调节能力。hyperperin可能对人类免疫缺陷病毒(HIV)和大多数革兰氏阴性细菌没有活性。然而,它对革兰氏阳性菌的抑菌活性和增强革兰氏阳性菌和革兰氏阴性菌的吞噬作用,可以保护获得性免疫缺陷综合征(AIDS)患者和其他免疫功能低下患者;一些革兰氏阴性菌,如奈瑟菌,对超穿孔素敏感。hyperperin具有穿透血脑屏障(BBB)和血睾丸屏障(BTB)的能力,是抗脑膜炎和淋病的宝贵抗菌剂。正如Bekondi等人(2006,Int.)最近所表明的那样,hyperperin的这些特性对于具有免疫调节活性的抗生素在与机会性细菌导致的艾滋病患者死亡率上升的斗争中是重要的。j .感染。生物学报,10,387-395)。它还可以帮助对抗与成人脑膜炎相关的原发性和机会性病原体与艾滋病毒血清状态的关系。
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引用次数: 7
More precise model of α-helix and transmembrane α-helical peptide backbone structure 更精确的α-螺旋和跨膜α-螺旋肽主链结构模型
Pub Date : 2012-11-29 DOI: 10.4236/JBPC.2012.34036
W. Schmidt, Clayton Thomas
The 3-D structure of the β-adrenergic receptor with a molecular weight of 55,000 daltons is available from crystallographic data. Within one of the seven transmembrane ion channel helices in the β2-receptor, one loop of a helix ACADL has previously been proposed as the site that explains β2 activity (fights acute bronchitis) whereas ASADL in the β1-receptor at the corresponding site explains β1-activity (cardiac stimulation). The α-agonist responsible for this selective reaction is only 0.5% of the receptor molecular weight, and only 1.5% of the weight of the trans-membrane portion of the receptor. The understanding of the mechanism by which a small molecule on binding to a site on one single loop of a helix produces a specific agonist activity on an entire transmembrane ion channel is uncertain. A model of an α-helix is presented in which of pitch occurs at angles both smaller and larger than 180° n. Consequently, atomic coordinates in a peptide backbone α-helix match the data points of individual atom (and atom types) in the backbone. More precisely, eleven atoms in peptide backbone routinely equal one loop of a helix, instead of eleven amino acid residues equaling three loops of a helix; therefore, an α-helix can begin (or end) at any specific atom in a peptide backbone, not just at any specific amino acid. Wavefront Topology System and Finite Element Methods calculate this specific helical shape based only upon circumference, pitch, and phase. Only external forces which specifically affect circumference, pitch and/or phase (e.g. from agonist binding) can/will alter the shape of an α-helix.
从晶体学数据可以得到分子量为55000道尔顿的β-肾上腺素能受体的三维结构。在β2受体的七个跨膜离子通道螺旋中的一个,螺旋ACADL的一个环先前被认为是解释β2活性(对抗急性支气管炎)的位点,而β1受体中相应位点的ASADL解释β1活性(心脏刺激)。负责这种选择性反应的α-激动剂仅占受体分子量的0.5%,仅占受体跨膜部分重量的1.5%。小分子与螺旋单环上的一个位点结合在整个跨膜离子通道上产生特异性激动剂活性的机制尚不确定。本文提出了一种α-螺旋的模型,其中螺距发生在小于和大于180°n的角度。因此,肽主链α-螺旋的原子坐标与主链中单个原子(和原子类型)的数据点相匹配。更准确地说,肽主链上的11个原子通常等于螺旋的一个环,而不是11个氨基酸残基等于螺旋的三个环;因此,α-螺旋可以在肽主链的任何特定原子上开始(或结束),而不仅仅是在任何特定的氨基酸上。波前拓扑系统和有限元方法计算这种特定的螺旋形状仅基于周长,节距和相位。只有特别影响周长、节距和/或相的外力(例如激动剂结合)才能/将改变α-螺旋的形状。
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引用次数: 6
26S proteasome inhibitors inhibit dexamethasone-dependent increase of tyrosine aminotransferase and tryptophan 2,3-dioxygenase mRNA levels in primary cultured rat hepatocytes 26S蛋白酶体抑制剂抑制原代培养大鼠肝细胞酪氨酸转氨酶和色氨酸2,3-双加氧酶mRNA水平的地塞米松依赖性升高
Pub Date : 2012-11-29 DOI: 10.4236/JBPC.2012.34043
M. Harashima, M. Hyuga, Y. Nagaoka, C. Saito, M. Furukawa, T. Seki, T. Ariga, N. Kawasaki, S. Niimi
Dexamethasone (Dex), a ligand for transcriptional enhancement of tyrosine aminotransferase (TAT) and tryptophan 2,3-dioxygenase (TO) genes, (100 nM) maximally increased these mRNA levels at 12 h and 7 h in primary cultured rat hepatocytes and the nuclear fraction, respectively. Lactacystin (5 μM) and epoxomicin (0.5 μM), 26S proteasome inhibitors, significantly suppressed the Dex-dependent maximum increase of TAT and TO mRNA levels in the cells and the nuclear fraction. Electrophoretic mobility shift assay demonstrated that lactacystin did not affect binding of glucocorticoid receptor to glucocorticoid responsive element. Furthermore, lactacystin did not affect the activation of GRE luciferase reporter by Dex transfected to the cells. The results demonstrate that 26S proteasome is positively involved in the Dex-dependent increase of TAT and TO mRNA levels in the cells and suggest that the mechanism of action of 26S proteasome may be degradation of some RNase(s), which breaks down TAT and TO mRNAs.
地塞米松(Dexamethasone, Dex)是一种增强酪氨酸氨基转移酶(TAT)和色氨酸2,3-双加氧酶(TO)基因转录的配体,(100 nM)在原代培养的大鼠肝细胞和核部分中分别在12 h和7 h最大限度地提高了这些mRNA的水平。26S蛋白酶体抑制剂Lactacystin (5 μM)和epoxomicin (0.5 μM)显著抑制了细胞和细胞核中TAT和TO mRNA水平的dex依赖性最大增幅。电泳迁移率转移实验表明,乳糖酵素不影响糖皮质激素受体与糖皮质激素反应元件的结合。此外,乳酸菌素不影响经Dex转染细胞的GRE荧光素酶报告基因的激活。结果表明,26S蛋白酶体积极参与细胞中TAT和TO mRNA水平的dex依赖性增加,并提示26S蛋白酶体的作用机制可能是降解一些分解TAT和TO mRNA的RNase。
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引用次数: 0
Genetic diversity of color phenotypes in the koi (Cyprinus carpio L.) as identified by molecular markers 鲤(Cyprinus carpio L.)颜色表型遗传多样性的分子标记鉴定
Pub Date : 2012-08-28 DOI: 10.4236/JBPC.2012.33029
Bercovich Dani, K. Sigal, Shauder Lior, Degani Gad
The common carp (Cyprinus carpio) has a large variety of strains. The more popular are the koi (Japanese ornamental carp), which are still bred today to generate creative colors and patterns, giving rise to multiple phenotypes. Since koi are in great demand, there is a challenge to determine the genetics defining their quality. Two methods: 1) direct sequencing of five candidate gene regions, i.e., mitochondrial (cytochrome b, 12S gene and the D-loop) and nuclear (red sensitive opsin and Rag-1) loci, to detect single nucleotide polymorphisms (SNP)s and 2) random amplification of polymorphic DNA (RAPD), were used to differentiate among four koi strains (Kohaku, Sanke, Ghost and Ohgon) and the common carp. Novel SNPs, distinguishing between koi and the common carp, were revealed in cytochrome b, the D-loop and in the red sensitive opsin; one was a missense mutation in cytochrome b at position 15860, in which threonine in the common carp became alanine in all koi strains examined. The Kohaku strain was found to have two alleles in the mitochondrial fragments, forming two different haplotypes (subpo-pulations). These novel SNPs distinguished between koi strains and the common carp, and the RAPD method enabled further differentiation among the four koi strains.
鲤鱼(Cyprinus carpio)有很多品种。更受欢迎的是锦鲤(日本观赏鱼),今天仍在繁殖,以产生创造性的颜色和图案,产生多种表型。由于锦鲤需求量很大,因此确定决定其质量的遗传因素是一个挑战。采用线粒体(细胞色素b、12S基因和D-loop)和细胞核(红色敏感视蛋白和rag1) 5个候选基因区直接测序检测单核苷酸多态性(SNP)s和随机扩增多态性DNA (RAPD)两种方法对Kohaku、Sanke、Ghost和Ohgon 4个锦鲤品系与普通鲤鱼进行区分。在细胞色素b、d -环和红色敏感视蛋白中发现了区分锦鲤和鲤鱼的新snp;一个是细胞色素b在15860位的错义突变,在所有的锦鲤品系中,普通鲤鱼的苏氨酸变成了丙氨酸。发现Kohaku菌株在线粒体片段中有两个等位基因,形成两种不同的单倍型(传票种群)。这些新的snp可以区分锦鲤品系和普通鲤鱼,RAPD方法可以进一步区分锦鲤品系。
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引用次数: 5
Quantitative measurement of p53-p53 antibody interactions by quartz crystal microbalance: A modelsystem for immunochemical calibration 用石英晶体微天平定量测量p53-p53抗体相互作用:一种免疫化学校准模型系统
Pub Date : 2012-08-28 DOI: 10.4236/JBPC.2012.33024
D. Atha, V. Reipa
We are developing methods to quantify antibody interactions that include a quartz crystal microbalance (QCM) system to measure, on a molecular basis, the interaction of p53 and anti-p53 antibodies. Previously, as a model system, we developed a measurement device consisting of p53 protein (human wild type), characterized by mass spectroscopy and immobilized at various concentrations on a glass slide. The device is designed as a control to be used with immunohistochemical (IHC) assays that incorporate commercially available anti-p53 antibodies and probes. In the current study, p53 protein is covalently immobilized on a silicon dioxide-coated quartz crystal and the resonance frequency shift is followed in-situ. The functionalized sensor is then incubated with the anti-p53 antibody, which provides a direct gravimetric measure of the antibody-antigen binding. This previously described method allows the comparison of the surface immobilized protein concentrations with estimates obtained by fluorescence measurement. The p53 functionalized QCM system offers an independent measure of surface immobilized protein concentration and is essential in development of our IHC calibration prototypes. These results provide a benchmark for comparing antibody systems that may be used in developing other molecular diagnostic assays such as immunocytochemical analysis and the detection of biomarker proteins in blood and urine.
我们正在开发定量抗体相互作用的方法,包括石英晶体微天平(QCM)系统,以分子为基础测量p53和抗p53抗体的相互作用。在此之前,我们开发了一个由p53蛋白(人类野生型)组成的测量装置作为模型系统,通过质谱表征并固定在不同浓度的玻璃载玻片上。该设备被设计为对照,用于免疫组织化学(IHC)测定,其中包括市售的抗p53抗体和探针。本研究将p53蛋白共价固定在二氧化硅包覆的石英晶体上,原位进行共振频移。功能化的传感器然后与抗p53抗体孵育,这提供了抗体-抗原结合的直接重量测量。这种先前描述的方法可以将表面固定的蛋白质浓度与荧光测量获得的估计进行比较。p53功能化的QCM系统提供了一种独立的表面固定蛋白浓度测量方法,在IHC校准原型的开发中至关重要。这些结果为比较抗体系统提供了一个基准,可用于开发其他分子诊断分析,如免疫细胞化学分析和血液和尿液中生物标记蛋白的检测。
{"title":"Quantitative measurement of p53-p53 antibody interactions by quartz crystal microbalance: A modelsystem for immunochemical calibration","authors":"D. Atha, V. Reipa","doi":"10.4236/JBPC.2012.33024","DOIUrl":"https://doi.org/10.4236/JBPC.2012.33024","url":null,"abstract":"We are developing methods to quantify antibody interactions that include a quartz crystal microbalance (QCM) system to measure, on a molecular basis, the interaction of p53 and anti-p53 antibodies. Previously, as a model system, we developed a measurement device consisting of p53 protein (human wild type), characterized by mass spectroscopy and immobilized at various concentrations on a glass slide. The device is designed as a control to be used with immunohistochemical (IHC) assays that incorporate commercially available anti-p53 antibodies and probes. In the current study, p53 protein is covalently immobilized on a silicon dioxide-coated quartz crystal and the resonance frequency shift is followed in-situ. The functionalized sensor is then incubated with the anti-p53 antibody, which provides a direct gravimetric measure of the antibody-antigen binding. This previously described method allows the comparison of the surface immobilized protein concentrations with estimates obtained by fluorescence measurement. The p53 functionalized QCM system offers an independent measure of surface immobilized protein concentration and is essential in development of our IHC calibration prototypes. These results provide a benchmark for comparing antibody systems that may be used in developing other molecular diagnostic assays such as immunocytochemical analysis and the detection of biomarker proteins in blood and urine.","PeriodicalId":62927,"journal":{"name":"生物物理化学(英文)","volume":"3 1","pages":"211-220"},"PeriodicalIF":0.0,"publicationDate":"2012-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"70901569","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 8
Visualization of decreased docosahexaenoic acid in the hippocampus of rats fed an n – 3 fatty acid-deficient diet by imaging mass spectrometry 用成像质谱法观察喂食n - 3脂肪酸缺乏饮食的大鼠海马中二十二碳六烯酸的减少
Pub Date : 2012-08-28 DOI: 10.4236/JBPC.2012.33025
S. Taira, M. Hashimoto, Kazunori Saito, O. Shido
The present study employed an imaging mass spectrometry (IMS) method to evaluate the effect of dietary n – 3 fatty acids on the fatty acid composition in rat brain. Rats were divided into two groups and fed either an n – 3 fatty acid-deficient or adequate diet. We determined the decreased n – 3 fatty acids in the hippocampus of rats fed an n – 3 fatty acid-deficient diet compared to the control. IMS visualization was achieved at a resolution of 100 m in rat brain, and showed decreased docosahexaenoic acid (DHA)-containing phosphatidyl choline (PC) or phosphatidyl ethanolamine (PE) in the hippocampus of rats fed an n – 3 fatty acid-deficient diet.
本研究采用成像质谱法(IMS)研究了饲粮中n - 3脂肪酸对大鼠脑内脂肪酸组成的影响。将大鼠分为两组,分别饲喂缺乏n - 3脂肪酸和充足的饮食。我们测定了与对照组相比,喂食n - 3脂肪酸缺乏饮食的大鼠海马中n - 3脂肪酸的减少。以100 m分辨率在大鼠脑内进行IMS可视化,结果显示,饲喂n - 3脂肪酸缺乏饮食的大鼠海马中含有二十二碳六烯酸(DHA)的磷脂酰胆碱(PC)或磷脂酰乙醇胺(PE)减少。
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引用次数: 8
期刊
生物物理化学(英文)
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