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Rat small intestine absorption and membrane digestion in the process of aging 大鼠衰老过程中小肠吸收和膜消化的变化
Pub Date : 2013-05-24 DOI: 10.4236/JBPC.2013.42009
S. T. Metelsky
Recently, in our experiments, we used the short-circuit current technique to study the kinetic constants for nutrient transporters in rat gastric-intestinal tract and the thickness of the intestinal unstirred layer near the mucosa surface. It was shown that, during the process of aging, the number of nutrient monomer transporters in the small intestine increases twofold, whereas the affinity of transporters to the correspondent nutrients remains unchanged. The situation for peptides may be opposite. The layer thickness in the vicinity of the mucosa surface, measured through glucose, decreased during the process of aging. It was suggested that, in old rats, the role of the digestive volume is more important, which results in an increase of the number of nutrient monomer transporters.
最近,我们在实验中采用短路电流技术研究了大鼠胃肠道营养转运体的动力学常数和近粘膜表面肠道未搅拌层的厚度。结果表明,在衰老过程中,小肠中营养物质单体转运蛋白的数量增加了两倍,而转运蛋白对相应营养物质的亲和力保持不变。多肽的情况可能正好相反。通过葡萄糖测量的粘膜表面附近的层厚在老化过程中下降。由此可见,在老龄大鼠中,消化容量的作用更为重要,从而导致营养单体转运体数量的增加。
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引用次数: 1
Biomonitoring of organic and inorganic chemicals in the hair of Vietnamese people via spectral and chromatographic analysis 通过光谱和色谱分析对越南人头发中的有机和无机化学物质进行生物监测
Pub Date : 2013-02-28 DOI: 10.4236/JBPC.2013.41001
L. P. Cuong, L. Thuy, M. Evgenev
A combination of spectral and chromatographic analytical methods has been executed to monitor chemicals in the hair of Vietnamese people. Methods were developed for the determination of some chemical elements in hair by inductively coupled-plasma atomic emission spectroscopy with limits of detection reaching 1 - 100 μg·l-1. A supercritical fluid extraction (SFE) and clean-up procedures were established for isolating organochlorine pollutants (OCPs) and organophosphorus pesticides (OPPs) from the hair of the Vietnamese test subjects. Eight OCPs and OPPs were determined by SFE with carbon dioxide only and methanol-modified carbon dioxide extraction at 350 bar and 45℃, followed by gas chromatography coupled to mass spectrometry. The results indicated that OCPs can be extracted quantitatively using CO2 only, whereas OPPs require a modifier for extraction. The limits of detection that were found were between 0.7 to 1.3 μg·g-1, and good linearity (r2 > 0.9861) was achieved within the tested ranges. The activities of the N-acetyltransferase (NAT) and the monooxygenases cytochrome P450 (CYP 450) were determined and compared to the acetylation and oxidation phenotypes of healthy Vietnamese test subjects. The variations in the concentration of chemical elements in human hair were shown to be dependent upon the activities of the two-phase metabolic system, on the individual’s gender and the duration of residence within Russia.
采用光谱和色谱相结合的分析方法来监测越南人头发中的化学物质。建立了电感耦合等离子体原子发射光谱法测定毛发中部分化学元素的方法,检出限为1 ~ 100 μg·l-1。建立了从越南受试者头发中分离有机氯污染物(OCPs)和有机磷农药(OPPs)的超临界流体萃取(SFE)和清理程序。在45℃、350 bar条件下,采用甲醇修饰的二氧化碳萃取法和SFE法分别测定8种OCPs和OPPs,然后采用气相色谱-质谱联用。结果表明,CO2可以定量提取OCPs,而OPPs需要改性剂才能提取。检测限在0.7 ~ 1.3 μg·g-1之间,在检测范围内线性良好(r2 > 0.9861)。测定了n -乙酰转移酶(NAT)和单加氧酶细胞色素P450 (cyp450)的活性,并与健康越南人的乙酰化和氧化表型进行了比较。人类头发中化学元素浓度的变化取决于两相代谢系统的活动、个人的性别和在俄罗斯居住的时间。
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引用次数: 2
Correlation of docking energies with spectroscopic kinetic assays of potential xanthine oxidase substrates 对接能与潜在黄嘌呤氧化酶底物光谱动力学分析的相关性
Pub Date : 2013-02-28 DOI: 10.4236/JBPC.2013.41003
Amy L Stockert, Tarek M Mahfouz, B. Petersen, Oluwaseun L. Fakunmoju
Here we present a docking model that ranks compounds according to their potential effectiveness as a potential substrate or inhibitor. We utilize xanthine oxidase (XO), a multi-cofactor oxido-reductase which converts hypoxanthine to xanthine and xanthine to uric acid. During the reductive half reaction, electrons flow from the molybdopterin, to each of two Fe/S centers, and finally to FAD. During the oxidative half reaction, electrons are passed from the FAD to O2. Under ideal physiological conditions, this reduction of oxygen generates H2O2 and, under multiple turnover conditions, superoxide in amounts which is regulated by catalase and superoxide dismutase. Utilizing computer modeling predictions of the docking orientations and energies of a group of purine based structures was selected. Correlating computer estimations with steady state kinetic data, a rapid screening process for inhibittor prediction was highlighted. This method allows educated selection of likely inhibitors, thereby decreasing the time and supplies required to complete a traditional kinetic analysis screening. Results demonstrate the functionality and reliability of this method and have proven particularly useful in understanding binding orienttations or poses of each compound.
在这里,我们提出了一个对接模型,根据它们作为潜在底物或抑制剂的潜在有效性对化合物进行排序。我们利用黄嘌呤氧化酶(XO),一种多辅助因子氧化还原酶,将次黄嘌呤转化为黄嘌呤和黄嘌呤转化为尿酸。在半还原反应中,电子从钼精中流出,流向两个Fe/S中心,最后流向FAD。在氧化半反应中,电子从FAD传递到O2。在理想的生理条件下,这种氧的还原产生H2O2,在多种周转条件下,产生超氧化物,其数量由过氧化氢酶和超氧化物歧化酶调节。利用计算机模型预测了一组嘌呤基结构的对接方向和能量。将计算机估计与稳态动力学数据相结合,强调了抑制剂预测的快速筛选过程。该方法允许有根据地选择可能的抑制剂,从而减少完成传统动力学分析筛选所需的时间和供应。结果证明了该方法的功能性和可靠性,并被证明在理解每种化合物的结合取向或姿势方面特别有用。
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引用次数: 0
A study of the interaction of drugs with liposomes with isothermal titration calorimetry 用等温滴定量热法研究药物与脂质体的相互作用
Pub Date : 2013-02-28 DOI: 10.4236/JBPC.2013.41002
H. Osanai, T. Ikehara, S. Miyauchi, K. Shimono, Jun Tamogami, T. Nara, N. Kamo
Isothermal titration calorimetry (ITC) was applied to investigate the interaction of drugs with liposomes. Two types of titration are possible. One type is when the liposome suspension in the cell is titrated by aliquots of drug solution, and the other is when the drug and liposome solutions take the opposite roles. In this paper, we employed the latter type because the disturbance of liposomes may be minimal in this titration type. We derived an equation in which the accumulated heat-flow is expressed as a function of the added lipid concentration. In the derivation, the uniform binding model was used although there may be various binding sites. This equation contains a parameter n, the number of binding sites per lipid molecule. In addition, we derive the relation between the dissociation constant (Kd), partition coefficient (Pm) and n. Binding parameters such as Kd, n, the Gibbs energy change, enthalpy change and entropy change were estimated for ANS (1-anilino-8-naphtarenesulfonate), TPB (tetraphenylborate), amlodipine, nifedipine, amitriptyline, nortriptyline, imipramine, desipramine, propranolol, chlorpromazine, promethazine, miconazole, indomethacin, diclofenac and diflunisal. For some drugs, the enthalpy change was the major binding affinity instead of the classical hydrophobic interaction in which entropy takes the essential role. We proved an approximate rule that for drugs with smaller n (the number of binding sites per lipid molecule), the entropy change contributes more than the enthalpy change.
采用等温滴定量热法(ITC)研究了药物与脂质体的相互作用。有两种滴定方法。一种是用等分的药物溶液滴定细胞中的脂质体悬浮液,另一种是药物和脂质体溶液发挥相反的作用。在本文中,我们采用后一种类型,因为在这种滴定类型中脂质体的干扰可能是最小的。我们推导了一个方程,其中累积的热流表示为添加的脂质浓度的函数。在推导中,虽然可能存在不同的结合位点,但我们采用了统一的结合模型。这个方程包含一个参数n,即每个脂质分子的结合位点数目。此外,我们还得到了解离常数(Kd)、分配系数(Pm)和n之间的关系。估计了ANS(1-苯胺-8-萘苯磺酸盐)、TPB(四苯基硼酸盐)、氨氯地平、硝苯地平、阿米替林、去甲替林、丙咪嗪、地丙嗪、心得安、氯丙嗪、异丙嗪、咪康唑、吲哚美辛、双氯芬酸和双氟尼松的结合参数Kd、n、吉布斯能变、焓变和熵变。对于某些药物,焓变是主要的结合亲和力,而不是熵起主要作用的经典疏水相互作用。我们证明了一个近似规则,即对于具有较小n(每个脂质分子的结合位点数目)的药物,熵变的贡献大于焓变。
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引用次数: 14
Protein and lipid characterization of wheat roots plasma membrane damaged by Fe and H 2 O 2 using ATR-FTIR method 用ATR-FTIR法表征铁和h2o2对小麦根质膜的破坏
Pub Date : 2013-02-28 DOI: 10.4236/JBPC.2013.41004
Xin Zhao, Xiaojun Yang, Yong Shi, Guoxiong Chen, Xin‐rong Li
In plant cells the plasma membrane is a highly elaborated structure that functions as the point of exchange with adjoining cells, cell walls and the external environment. In this study, we investigated the structure and function characteristic of wheat root plasma membrane (PM) as affected by H2O2 and Fe by using fluorescence spectroscopic and attenuated total reflectance infrared (ATR-IR) techniques. The results showed that these oxidant damaged induced an obviously reduced membrane fluidity were observed in the roots PM treated with the 200 μM H2O2, FeSO4, and FeCl3. Computer-aided software analyses of the FTIR spectrum indicated that the content of the α-helices decreased and β-sheet increased in the secondary structures of proteins after exposure to the oxidants of 200 μM H2O2, FeSO4, and FeCl3. The number of P=O and C=C bonds area declined rapidly in the lipids of the membrane under the oxidants stress. These structural alterations might explain the reason of the roots PM instability under most of the abiotic stress.
在植物细胞中,质膜是一种高度复杂的结构,其功能是与相邻细胞、细胞壁和外部环境进行交换。本研究利用荧光光谱和衰减全反射红外(ATR-IR)技术研究了H2O2和Fe对小麦根质膜(PM)结构和功能的影响。结果表明,在200 μM H2O2、FeSO4和FeCl3处理下,这些氧化损伤导致根系PM膜流动性明显降低。计算机辅助软件FTIR分析表明,在200 μM H2O2、FeSO4和FeCl3的氧化作用下,蛋白质二级结构中α-螺旋的含量减少,β-片的含量增加。氧化胁迫下膜脂质中P=O和C=C键面积迅速减少。这些结构变化可能解释了在大多数非生物胁迫下根系PM不稳定的原因。
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引用次数: 8
Synthesis and characterization of hydroxyapatite contain chromium 含铬羟基磷灰石的合成与表征
Pub Date : 2012-11-29 DOI: 10.4236/JBPC.2012.34033
S. Sallam, Khairi M. Tohami, A. Sallam, L. I. A. Salem, F. A. Mohamed
Hydroxy apatite (HA) is a biocompatible material used as bone-substitute materials in both orthopedics and dentistry due to its excellent osteo-conductive and chemical composition similar to that of the inorganic part of bone in comparison with other implant materials . HA (S1) and Cr loaded hydroxy apatite (S2, S3, S4 and S5) of different chromium concentrations have been prepared. many techniques used to analyze the changes occurred due to the substitution. The XRD (X-ray diffraction) and FTIR (Fourier transform infrared radiation )spectroscopy were used as analytical techniques for the prepared HA and Cr loaded HA. The effect of the heat treatment on structural change of the samples was examined by DTA. It was found that all the samples have the same crystalline structure. The crystallinity of the samples decreased as the chromium ions concentration increase.
羟基磷灰石(hydroxyapatite, HA)是一种生物相容性材料,与其他种植材料相比,具有优异的骨导电性和与骨无机部分相似的化学成分,是骨科和牙科中用作骨替代材料的生物相容性材料。制备了不同铬浓度的羟基磷灰石(S1)和载Cr羟基磷灰石(S2、S3、S4和S5)。许多用于分析由于替换而发生的变化的技术。采用XRD (x射线衍射)和FTIR(傅里叶变换红外辐射)光谱对制备的HA和载Cr的HA进行了分析。采用差热分析(DTA)研究了热处理对试样组织变化的影响。结果表明,所有样品具有相同的晶体结构。随着铬离子浓度的增加,样品的结晶度降低。
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引用次数: 25
A single base permutation in any loop of a folded intramolecular quadruplex influences its structure and stability 在折叠的分子内四联体的任何环中,单碱基排列都会影响其结构和稳定性
Pub Date : 2012-11-29 DOI: 10.4236/JBPC.2012.34042
D. Yadav, R. Sheardy
The human telomere sequence (TTAGGG)4 folds into an unusual conformation possessing three G-tetrads linked by TTA loops. The first loop is a propeller loop while the second and third loops are transverse loops. Using Circular Dichroism (CD) spectroscopy, we have investigated the effect of sequence context on the structures and stabilities of intramolecular G-quadruplexes related to the human telomere sequence by considering all permutations of T and A within the loops. The results indicate that changing only one base in any one loop can have a dramatic effect on the conformation of the quadruplex as well as its melting temperature, Tm. Thus, each sequence studied has a unique CD spectrum and Tm. In general, variants with a modified second loop are the most stable while the wild type sequence is the least stable. The observed difference in CD spectra and melting temperature are discussed in terms of base stacking within the loop and stacking of the loop bases with adjacent G-tetrads.
人类端粒序列(TTAGGG)4折叠成一个不寻常的构象,具有三个由TTA环连接的g四分体。第一个环是螺旋桨环,而第二个和第三个环是横向环。利用圆二色(CD)光谱,我们通过考虑环内T和A的所有排列,研究了序列背景对与人类端粒序列相关的分子内g -四联体结构和稳定性的影响。结果表明,只要改变任一环中的一个碱基,就能对四联体的构象及其熔融温度产生显著影响。因此,所研究的每个序列都具有独特的CD谱和Tm。一般来说,具有修改的第二环的变体是最稳定的,而野生型序列是最不稳定的。从环内基的叠加和环内基与相邻g四分体的叠加的角度讨论了所观察到的CD光谱和熔化温度的差异。
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引用次数: 6
In vivo effects of genistein, herbimycin a and geldanamycin on rat hepatic cytochrome P4501A 染料木素、红霉素a和格尔达霉素对大鼠肝细胞色素P4501A的体内影响
Pub Date : 2012-11-29 DOI: 10.4236/JBPC.2012.34041
M. Perepechaeva, A. Grishanova
Cytochrome P4501A (the CYP1A1 and CYP1A2 enzymes) are regulated through the aryl hydrocarbon receptor (AhR)-dependent signal transduction pathway and are generally known as enzymes which metabolize anthropogenic xenobiotics such as dioxin to carcinogenic and mutagenic compounds. However, recently the facts of CYP1A activation under physiological conditions or under action of non-dioxin-like compounds appear. In the present study we show that genistein, herbimycin A and geldanamycin (the protein-tyrosine kinase inhibitors) affect in vivo to CYP1A1 activity, the CYP1A1 mRNA level and the CYP1A1 protein level. These data provide insight into the role of protein kinases in CYP1A regulation may facilitate the understanding of CYP1A regulation.
细胞色素P4501A (CYP1A1和CYP1A2酶)是通过芳烃受体(AhR)依赖的信号转导途径调节的,通常被称为将二恶英等人为外源物代谢为致癌和致突变化合物的酶。然而,最近CYP1A在生理条件下或在非二恶英类化合物的作用下激活的事实出现了。在本研究中,我们发现染料木素、herbimycin A和格尔达霉素(蛋白酪氨酸激酶抑制剂)在体内影响CYP1A1活性、CYP1A1 mRNA水平和CYP1A1蛋白水平。这些数据提供了对蛋白激酶在CYP1A调节中的作用的深入了解,可能有助于理解CYP1A的调节。
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引用次数: 4
Protective effect of fermented papaya preparation on stress-induced acute gastric mucosal lesion 发酵木瓜制剂对应激性急性胃黏膜损伤的保护作用
Pub Date : 2012-11-29 DOI: 10.4236/JBPC.2012.34038
S. Murakami, F. Takayama, T. Egashira, Mitsuko Imao, A. Mori
Stress ulceration is single or multiple mucosal defects with/without bleeding from the gastric mucosa during the physiologic stress. Oxidative stress (OS) is a key pathogenic factor in psychogenic stress-induced acute gastric mucosal lesion (AGML). Fermented papaya preparation (FPP) is reported to have oxygen radical scavenging activity and is effective in OS-related diseases. Here, we investigated the protective effects and the mechanism of action of FPP on stress-induced AGML in rats, induced by water immersion restraint stress (WIRS). Exposure of rats to 6-hour WIRS resulted in the appearance of splinter hemorrhages and mucosal lesions in the stomach. WIRS induced significant increase in lipid peroxidation and decrease in superoxide dismutase-like activity in both the plasma and gastric mucosa. WIRS also significantly increased myeloperoxidase activity together with Nuclear factor-kappaB (NF-kB) activation in gastric mucosa. FPP reduced all the above changes. The results suggest that oral administration of FPP provides protection against WIRS-induced acute gastric mucosal lesions through its anti-oxidative and anti-inflammatory properties.
应激性溃疡是指在生理性应激作用下,胃粘膜出现单个或多个粘膜缺损,伴有或不伴有出血。氧化应激(Oxidative stress, OS)是心因性应激引起的急性胃粘膜病变(AGML)的重要致病因素。据报道,发酵木瓜制剂(FPP)具有清除氧自由基的活性,对os相关疾病有效。本研究探讨了FPP对水浸抑制应激(WIRS)大鼠应激性AGML的保护作用及其机制。将大鼠暴露于6小时的WIRS导致胃出现碎片状出血和粘膜病变。WIRS诱导血浆和胃粘膜脂质过氧化显著增加,超氧化物歧化酶样活性显著降低。WIRS还显著增加了胃粘膜髓过氧化物酶活性和核因子κ b (NF-kB)的激活。FPP降低了上述所有变化。结果表明,口服FPP通过其抗氧化和抗炎作用对wirs诱导的急性胃黏膜病变具有保护作用。
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引用次数: 10
Microwave based reversible unfolding and refolding of alcohol oxidase protein probed by fluorescence and circular dichroism spectroscopy 用荧光和圆二色光谱研究微波基酒精氧化酶蛋白的可逆展开和再折叠
Pub Date : 2012-11-29 DOI: 10.4236/JBPC.2012.34039
Somasekhar R. Chinnadayyala, M. Santhosh, P. Goswami
The reversible effect of microwave mediated denaturation of protein at low exposure time of 10 s has been demonstrated for the first time. The effect of microwave (2.45 GHz and 900 W) was confirmed in a homo-octameric alcohol oxidase in aqueous solution of pH 7.5. The unfolding events did not transverse through any intermediate states and no subunits of the protein were detached during the process. The refolding of the protein achieved at 4℃ for 24 h had regenerated the native enzyme. This reversible refolding approach excludes any chemical reagent and therefore established as simple technique for protein unfolding-folding studies.
本文首次证实了微波介导蛋白质在低暴露时间10s下变性的可逆效应。在pH为7.5的水溶液中,证实了微波(2.45 GHz, 900 W)对同型八聚醇氧化酶的影响。在此过程中,展开事件没有横向通过任何中间状态,也没有蛋白质的亚基分离。在4℃、24 h条件下对蛋白质进行再折叠,使原酶再生。这种可逆的再折叠方法排除了任何化学试剂,因此被确立为蛋白质展开折叠研究的简单技术。
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引用次数: 4
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生物物理化学(英文)
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