• 【天然非结构化α-突触核蛋白向其α-螺旋构象的转化显着减弱了活性氧的产生。】 复制标题 收藏 收藏
    DOI:10.1016/j.jinorgbio.2012.09.001 复制DOI
    作者列表:Zhou B,Hao Y,Wang C,Li D,Liu YN,Zhou F
    BACKGROUND & AIMS: :The intracellular α-synuclein (α-syn) protein, whose conformational change and aggregation have been closely linked to the pathology of Parkingson's disease (PD), is highly populated at the presynaptic termini and remains there in the α-helical conformation. In this study, circular dichroism confirmed that natively unstructured α-syn in aqueous solution was transformed to its α-helical conformation upon addition of trifluoroethanol (TFE). Electrochemical and UV-visible spectroscopic experiments reveal that both Cu (I) and Cu (II) are stabilized, with the former being stabilized by about two orders of magnitude. Compared to unstructured α-syn (Binolfi et al., J. Am. Chem. Soc. 133 (2011) 194-196), α-helical α-syn stabilizes Cu (I) by more than three orders of magnitude. Through the measurements of H(2)O(2) and hydroxyl radicals (OH) in solutions containing different forms of Cu (II) (free and complexed by unstructured or α-helical α-syn), we demonstrate that the significantly enhanced Cu (I) binding affinity helps inhibit the production of highly toxic reactive oxygen species, especially the hydroxyl radicals. Our study provides strong evidence that, as a possible means to prevent neuronal cell damage, conversion of the natively unstructured α-syn to its α-helical conformation in vivo could significantly attenuate the copper-modulated ROS production.
    背景与目标: :胞内α-突触核蛋白(α-syn)蛋白的构象变化和聚集与帕金森氏病(PD)的病理状况密切相关,在突触前末端高密度分布,并保持在α-螺旋构象中。在这项研究中,圆二色性证实,加入三氟乙醇(TFE)后,水溶液中的天然非结构化α-syn转变为其α-螺旋构象。电化学和紫外可见光谱实验表明,Cu(I)和Cu(II)都稳定了,前者稳定了大约两个数量级。与非结构化α-syn相比(Binolfi等人,J.Am.Chem.Soc.133(2011)194-196),α-螺旋α-syn使Cu(I)稳定超过三个数量级。通过测量H(2)O(2)和羟基自由基(OH)在包含不同形式的Cu(II)(游离且由非结构化或α-螺旋α-syn络合)的溶液中,我们证明了显着增强的Cu (I)结合亲和力有助于抑制剧毒活性氧的产生,特别是羟基自由基的产生。我们的研究提供了有力的证据,作为一种可能的预防神经元细胞损伤的方法,体内天然非结构化的α-syn转化为其α-螺旋构象可以显着减弱铜调节的ROS产生。
  • 【煤诱导了斯氏假单胞菌生产鼠李糖脂生物表面活性剂,该假单胞菌从Jharia煤层的地层水中分离出来。】 复制标题 收藏 收藏
    DOI:10.1016/j.biortech.2012.10.127 复制DOI
    作者列表:Singh DN,Tripathi AK
    BACKGROUND & AIMS: :A strain of Pseudomonas stutzeri was isolated form an enrichment of perchlorate reducing bacteria from the formation water collected from an Indian coalbed which solubilized coal and produced copious amount of biosurfactant when coal was added to the medium. It produced maximum biosurfactant with lignite coal followed by olive oil and soybean oil which was able to emulsify several aromatic hydrocarbons including kerosene oil, diesel oil, hexane, toluene etc. Haemolytic test, growth inhibition of Bacillus subtilis and FTIR analysis showed rhamnolipid nature of the biosurfactant. The stability of the coal induced biosurfactant in pH range of 4-8 and up to 25% NaCl concentration and 100 °C temperature suggests that due to its ability to produce biosurfactant and solubilize coal P. stutzeri may be useful in the coalbed for in situ biotransformation of coal into methane and in the bioremediation of PAHs from oil contaminated sites including marine environments.
    背景与目标: :从从印度煤层中收集的地层水中分离出富集高氯酸盐的细菌,分离出一株假单胞菌假单胞菌,当将煤添加到培养基中时,该煤层溶解了煤炭并产生了大量生物表面活性剂。它生产的生物表面活性剂最多的是褐煤,其次是橄榄油和大豆油,能够乳化煤油,柴油,己烷,甲苯等几种芳烃。生物表面活性剂。煤炭诱导的生物表面活性剂在4-8的pH范围内以及最高25%的NaCl浓度和100°C的温度下的稳定性表明,由于其具有产生生物表面活性剂和溶解煤炭的能力,斯图哲木可能在原位用于煤层中。煤的生物转化为甲烷,以及在石油污染的地点(包括海洋环境)中对多环芳烃的生物修复。
  • 【用于第二代生物燃料生产的超基因组功能纤维素酶的生物勘探:综述。】 复制标题 收藏 收藏
    DOI:10.1080/1040841X.2017.1337713 复制DOI
    作者列表:Tiwari R,Nain L,Labrou NE,Shukla P
    BACKGROUND & AIMS: :Second generation biofuel production has been appeared as a sustainable and alternative energy option. The ultimate aim is the development of an industrially feasible and economic conversion process of lignocellulosic biomass into biofuel molecules. Since, cellulose is the most abundant biopolymer and also represented as the photosynthetically fixed form of carbon, the efficient hydrolysis of cellulose is the most important step towards the development of a sustainable biofuel production process. The enzymatic hydrolysis of cellulose by suites of hydrolytic enzymes underlines the importance of cellulase enzyme system in whole hydrolysis process. However, the selection of the suitable cellulolytic enzymes with enhanced activities remains a challenge for the biorefinery industry to obtain efficient enzymatic hydrolysis of biomass. The present review focuses on deciphering the novel and effective cellulases from different environmental niches by unculturable metagenomic approaches. Furthermore, a comprehensive functional aspect of cellulases is also presented and evaluated by assessing the structural and catalytic properties as well as sequence identities and expression patterns. This review summarizes the recent development in metagenomics based approaches for identifying and exploring novel cellulases which open new avenues for their successful application in biorefineries.
    背景与目标: :第二代生物燃料生产已被视为一种可持续的替代能源选择。最终目的是开发一种工业上可行且经济的木质纤维素生物质转化为生物燃料分子的方法。由于纤维素是最丰富的生物聚合物,并且也表现为碳的光合作用固定形式,因此纤维素的有效水解是朝着可持续生物燃料生产工艺发展的最重要步骤。一系列水解酶对纤维素的酶促水解强调了纤维素酶系统在整个水解过程中的重要性。然而,选择具有增强活性的合适的纤维素分解酶对于生物精炼工业获得有效的生物质的酶水解仍然是一个挑战。目前的审查侧重于通过不可培养的宏基因组学方法从不同的环境利基破译新颖和有效的纤维素酶。此外,还介绍了纤维素酶的综合功能方面,并通过评估结构和催化特性以及序列同一性和表达模式来对其进行评估。这篇综述总结了基于宏基因组学的方法的最新发展,该方法用于鉴定和探索新型纤维素酶,为它们在生物精炼厂的成功应用开辟了新途径。
  • 【Cissampelos sympodialis Eichl叶片的乙醇提取物的水相部分的支气管扩张活性。 (Menispermaceae)在豚鼠中。】 复制标题 收藏 收藏
    DOI:10.1016/S0944-7113(97)80073-6 复制DOI
    作者列表:Thomas G,Araújo CC,Duarte JC,De Souza DP
    BACKGROUND & AIMS: :The bronchodilator activity of an aqueous fraction (AF) of a 70% hydroalcoholic extract of the leaves of Cissampelos sympodialis Eichl. was evaluated in the guinea-pig. The AF inhibited the spontaneous tone of the trachea (IC(50), 13.9 μg/ml), which was potentiated (IC(50), 4.6 μg/ml) by 3-isobutyl-l-methylxanthine, blocked by β(2) adrenoceptor blocking agent timolol, but unaffected by removal of epithelium or addition of NG-Nitro-L-arginine methyl ester or methylene blue. The AF also antagonized contractions induced by carbachol, capsaicin and arachidonic acid in normal trachea and by ovalbumin in trachea obtained from sensitized guinea-pigs. The IC(50)-values in these experiments varied from 34.1-70.5 μg/ml. Further, the AF (100 mg/kg) by the more effective i.p. route increased the preconvulsive time of animals exposed to an aerosol of histamine to 63.5 ± 5 s 1 h after administration compared to 28 ± 1 s in the untreated group. In addition, the AF at 100 mg/kg i.p. or i.G. protected the sensitized guinea-pigs against anaphylactic shock induced by ovalbumin aerosol by 78.6 and 86.7% respectively.
    背景与目标: :七叶蝉叶的70%的水醇提取物的水相(AF)的支气管扩张活性。在豚鼠中进行了评估。 AF抑制气管的自发音(IC(50),13.9μg/ ml),其被3-异丁基-1-甲基黄嘌呤增强并被β(2)阻断(IC(50),4.6μg/ ml)。肾上腺素受体阻滞剂噻吗洛尔,但不受上皮去除或添加NG-硝基-L-精氨酸甲酯或亚甲基蓝的影响。房颤还拮抗正常气管中卡巴胆碱,辣椒素和花生四烯酸引起的收缩,以及从致敏的豚鼠获得的气管中卵清蛋白引起的收缩。这些实验中的IC(50)值从34.1-70.5μg/ ml不等。此外,AF(100 mg / kg)的腹腔镜效果更佳。给药后1小时,暴露于组胺气溶胶的动物的惊厥前时间延长至63.5±5 s,而未治疗组为28±1 s。此外,AF的i.p.为100 mg / kg。或i.G.致敏豚鼠对卵白蛋白气溶胶引起的过敏性休克的保护作用分别为78.6和86.7%。
  • 【中间链球菌中溶血素产生的血液成分的正控制和负控制途径。】 复制标题 收藏 收藏
    DOI:10.1128/IAI.00379-17 复制DOI
    作者列表:Tomoyasu T,Yamasaki T,Chiba S,Kusaka S,Tabata A,Whiley RA,Nagamune H
    BACKGROUND & AIMS: :Streptococcus intermedius is an opportunistic bacterial pathogen secreting a human-specific cytolysin called intermedilysin (ILY) as a major pathogenic factor. This bacterium can degrade glycans into monosaccharides using two glycosidases, multisubstrate glycosidase A (MsgA) and neuraminidase (NanA). Here, we detected a stronger hemolytic activity mediated by ILY when S. intermedius PC574 was cultured in fetal bovine serum (FBS) than when it was grown in the standard culture medium. FBS-cultured cells also showed higher MsgA and NanA activity, although overproduction of ILY in FBS was undetectable in mutants nanA-null and msgA-null. Addition of purified MsgA and NanA to the FBS resulted in a release of 2.8 mM galactose and 4.3 mM N-acetylneuraminic acid; these sugar concentrations were sufficient to upregulate the expression of ILY, MsgA, and NanA. Conversely, when strain PC574 was cultured in human plasma, no similar increase in hemolytic activity was observed. Moreover, addition of human plasma to the culture in FBS appeared to inhibit the stimulatory effect of FBS on ILY, MsgA, and NanA, although there were individual differences among the plasma samples. We confirmed that human plasma contains immunoglobulins that can neutralize ILY, MsgA, and NanA activities. In addition, human plasma had a neutralizing effect on cytotoxicity of S. intermedius toward HepG2 cells in FBS, and a higher concentration of human plasma was necessary to reduce the cytotoxicity of an ILY-high-producing strain than an ILY-low-producing strain. Overall, our data show that blood contains factors that stimulate and inhibit ILY expression and activity, which may affect pathogenicity of S. intermedius.
    背景与目标: :Intermedococcus intermedius是一种机会细菌性病原体,会分泌称为intermedilysin(ILY)的人类特异性溶细胞素,这是主要的致病因素。该细菌可以使用两种糖苷酶,多底物糖苷酶A(MsgA)和神经氨酸酶(NanA)将聚糖降解为单糖。在这里,与在标准培养基中生长相比,在胎牛血清(FBS)中培养中间链霉菌PC574时,我们检测到由ILY介导的溶血活性更强。 FBS培养的细胞也显示出更高的MsgA和NanA活性,尽管在突变体nanA-null和msgA-null中无法检测到FLY中ILY的过量产生。向FBS中添加纯化的MsgA和NanA导致释放2.8 mM半乳糖和4.3 mM N-乙酰神经氨酸。这些糖浓度足以上调ILY,MsgA和NanA的表达。相反,当在人血浆中培养PC574菌株时,未观察到溶血活性的类似增加。此外,尽管血浆样品之间存在个体差异,但在FBS的培养物中添加人血浆似乎抑制了FBS对ILY,MsgA和NanA的刺激作用。我们证实人血浆中含有可以中和ILY,MsgA和NanA活性的免疫球蛋白。另外,人血浆对中间人链球菌对FBS中的HepG2细胞的细胞毒性具有中和作用,为了降低ILY高产菌株的细胞毒性,需要更高浓度的人血浆来降低ILY高产菌株的细胞毒性。 。总的来说,我们的数据表明血液中含有刺激和抑制ILY表达和活性的因子,这些因子可能影响中间链球菌的致病性。
  • 【通过减轻细胞表面工程酵母菌株对纤维素酶的不可逆吸附,可同时改善结晶纤维素的糖化作用和乙醇生产。】 复制标题 收藏 收藏
    DOI:10.1007/s00253-012-4587-x 复制DOI
    作者列表:Matano Y,Hasunuma T,Kondo A
    BACKGROUND & AIMS: :Enzymatic hydrolysis of cellulosic material is an essential step in the bioethanol production process. However, complete cellulose hydrolysis by cellulase is difficult due to the irreversible adsorption of cellulase onto cellulose. Thus, part of the cellulose remains in crystalline form after hydrolysis. In this study, after 96-h hydrolysis of Avicel crystalline cellulose, 47.1 % of the cellulase was adsorbed on the cellulose surface with 10.8 % crystalline cellulose remaining. In simultaneous saccharification and fermentation of 100 g/L Avicel with 1.0 filter paper unit/mL cellulase, a wild-type yeast strain produced 44.7 g/L ethanol after 96 h. The yield of ethanol was 79.7 % of the theoretical yield. On the other hand, a recombinant yeast strain displaying various cellulases, such as β-glucosidase, cellobiohydrolase, and endoglucanase, produced 48.9 g/L ethanol, which corresponds to 87.3 % of the theoretical yield. Higher ethanol production appears to be attributable to higher efficiency of cellulase displayed on the cell surface. These results suggest that cellulases displayed on the yeast cell surface improve hydrolysis of Avicel crystalline cellulose. Indeed, after the 96-h simultaneous saccharification and fermentation using the cellulase-displaying yeast, the amount of residual cellulose was 1.5 g/L, one quarter of the cellulose remaining using the wild-type strain, a result of the alleviation of irreversible adsorption of cellulases on the crystalline cellulose.
    背景与目标: :纤维素材料的酶水解是生物乙醇生产过程中必不可少的步骤。但是,由于纤维素酶不可逆地吸附在纤维素上,因此很难通过纤维素酶将纤维素完全水解。因此,部分纤维素在水解后保持为结晶形式。在这项研究中,Avicel结晶纤维素水解96小时后,47.1%的纤维素酶被吸附在纤维素表面,而剩余的10.8%结晶纤维素被吸收。在同时用100毫升滤纸单位/ mL纤维素酶糖化和发酵100 g / L Avicel的过程中,一种野生型酵母菌株在96小时后产生了44.7 g / L乙醇。乙醇的产率为理论产率的79.7%。另一方面,展示各种纤维素酶,例如β-葡萄糖苷酶,纤维二糖水解酶和内切葡聚糖酶的重组酵母菌株产生了48.9 g / L乙醇,相当于理论收率的87.3%。较高的乙醇产量似乎归因于细胞表面展示的纤维素酶效率更高。这些结果表明,展示在酵母细胞表面上的纤维素酶改善了Avicel结晶纤维素的水解。实际上,在使用展示纤维素酶的酵母糖进行96小时同步糖化和发酵后,残留纤维素的量为1.5 g / L,其中野生型菌株残留的纤维素的四分之一是减轻不可逆吸附的结果纤维素在结晶纤维素上的分布。
  • 【在空运生物反应器中大规模种植紫锥菊的不定根,用于生产Chichoricic酸,绿原酸和Caftaric酸。】 复制标题 收藏 收藏
    DOI:10.1007/s10529-007-9399-1 复制DOI
    作者列表:Wu CH,Murthy HN,Hahn EJ,Paek KY
    BACKGROUND & AIMS: :Adventitious roots of Echinacea purpurea were cultured in airlift bioreactors (20 l, 500 l balloon-type, bubble bioreactors and 1,000 l drum-type bubble bioreactor) using Murashige and Skoog (MS) medium with 2 mg indole butyric acid l(-1) and 50 g sucrose l(-1) for the production of chichoric acid, chlorogenic acid and caftaric acid. In the 20 l bioreactor (containing 14 l MS medium) a maximum yield of 11 g dry biomass l(-1) was achieved after 60 days. However, the amount of total phenolics (57 mg g(-1) DW), flavonoids (34 mg g(-1) DW) and caffeic acid derivatives (38 mg g(-1) DW) were highest after 50 days. Based on these studies, pilot-scale cultures were established and 3.6 kg and 5.1 kg dry biomass were achieved in the 500 l and 1,000 l bioreactors, respectively. The accumulation of 5 mg chlorogenic acid g(-1) DW, 22 mg chichoric acid g(-1) DW and 4 mg caftaric acids g(-1) DW were achieved with adventitious roots grown in 1,000 l bioreactors.
    背景与目标: :紫锥菊的次生根在装有2 mg吲哚丁酸l(-1)的Murashige和Skoog(MS)培养基中于气举生物反应器(20 l,500 l气球型,气泡生物反应器和1,000 l鼓型气泡生物反应器)中培养。 )和50克蔗糖l(-1)用于生产手性酸,绿原酸和异戊酸。 60天后,在20升生物反应器(包含14升MS培养基)中,最大产量为11克干燥生物质l(-1)。但是,总酚含量(57 mg g(-1)DW),类黄酮(34 mg g(-1)DW)和咖啡酸衍生物(38 mg g(-1)DW)的量在50天后最高。基于这些研究,建立了中试规模的培养,分别在500 l和1,000 l生物反应器中实现了3.6 kg和5.1 kg干燥生物量。 5 mg绿原酸g(-1)DW,22 mg chichoric acid g(-1)DW和4 mg caftaric acid g(-1)DW的积累是通过在1,000升生物反应器中生长的不定根实现的。
  • 【结核病患者皮质醇/脱氢表雄酮比值的变化及其与外周血单核细胞由分枝杆菌驱动的细胞因子产生异常的关系。】 复制标题 收藏 收藏
    DOI:10.1111/j.1365-3083.2007.01952.x 复制DOI
    作者列表:Bozza VV,D'Attilio L,Mahuad CV,Giri AA,Del Rey A,Besedovsky H,Bottasso O,Bay ML
    BACKGROUND & AIMS: :We have investigated the relationship between cortisol and dehydroepiandrosterone (DHEA) levels and the immune response to mycobacterial antigens in peripheral venous blood, from a male population of active tuberculosis patients and age-matched healthy controls of the same sex (HCo). Peripheral blood mononuclear cells were cultured for 36 or 96 h with whole sonicated Mycobacterium tuberculosis (WSA) for measurement of proliferation, interferon gamma (IFN-gamma) and interleukin-10 (IL-10) in culture supernatants. Comparisons on the in vitro mycobacterial-driven immune responses demonstrated that TB patients had a higher IL-10 production, a decreased lymphoproliferation and a trend to reduced IFN-gamma synthesis, in relation to HCo. Active disease was also characterized by increases in the plasma levels of glucocorticoids (GC) and reduced concentrations of DHEA which resulted in a higher cortisol/DHEA ratio respect the HCo group. Plasma DHEA levels were positively correlated with IFN-gamma values. An inverse correlation was found between the cortisol/DHEA ratio and IFN-gamma levels. Novel evidence is provided showing that the balance between cortisol and DHEA is partly responsible for the immune perturbations seen in TB patients.
    背景与目标: :我们研究了男性活跃结核病患者人群和年龄匹配的同性健康对照者(HCo)的皮质醇和脱氢表雄酮(DHEA)水平与外周静脉血对分枝杆菌抗原的免疫反应之间的关系。将外周血单核细胞与全超声分枝杆菌(WSA)培养36或96 h,以测量培养上清液中的增殖,干扰素γ(IFN-γ)和白介素10(IL-10)。体外分枝杆菌驱动的免疫反应的比较表明,与HCo相比,结核病患者具有更高的IL-10产生,降低的淋巴增殖和减少IFN-γ合成的趋势。活动性疾病的特征还在于血浆糖皮质激素(GC)水平升高和DHEA浓度降低,从而导致相对于HCo组更高的皮质醇/ DHEA比。血浆DHEA水平与IFN-γ值呈正相关。发现皮质醇/ DHEA比与IFN-γ水平成反比。提供了新的证据,表明皮质醇和DHEA之间的平衡是造成TB患者免疫紊乱的部分原因。
  • 【由过量表达的异源α-淀粉酶引起的生产和分泌应激导致枯草芽孢杆菌中的孢子形成受到抑制,并且活动期延长。】 复制标题 收藏 收藏
    DOI:10.1128/AEM.00472-07 复制DOI
    作者列表:Lulko AT,Veening JW,Buist G,Smits WK,Blom EJ,Beekman AC,Bron S,Kuipers OP
    BACKGROUND & AIMS: :Transcriptome analysis was used to investigate the global stress response of the gram-positive bacterium Bacillus subtilis caused by overproduction of the well-secreted AmyQ alpha-amylase from Bacillus amyloliquefaciens. Analyses of the control and overproducing strains were carried out at the end of exponential growth and in stationary phase, when protein secretion from B. subtilis is optimal. Among the genes that showed increased expression were htrA and htrB, which are part of the CssRS regulon, which responds to high-level protein secretion and heat stress. The analysis of the transcriptome profiles of a cssS mutant compared to the wild type, under identical secretion stress conditions, revealed several genes with altered transcription in a CssRS-dependent manner, for example, citM, ylxF, yloA, ykoJ, and several genes of the flgB operon. However, high-affinity CssR binding was observed only for htrA, htrB, and, possibly, citM. In addition, the DNA macroarray approach revealed that several genes of the sporulation pathway are downregulated by AmyQ overexpression and that a group of motility-specific (sigmaD-dependent) transcripts were clearly upregulated. Subsequent flow-cytometric analyses demonstrate that, upon overproduction of AmyQ as well as of a nonsecretable variant of the alpha-amylase, the process of sporulation is severely inhibited. Similar experiments were performed to investigate the expression levels of the hag promoter, a well-established reporter for sigmaD-dependent gene expression. This approach confirmed the observations based on our DNA macroarray analyses and led us to conclude that expression levels of several genes involved in motility are maintained at high levels under all conditions of alpha-amylase overproduction.
    背景与目标: :转录组分析用于研究革兰氏阳性枯草芽孢杆菌的总体应激反应,该酶是由解淀粉芽孢杆菌分泌的大量分泌良好的AmyQα-淀粉酶引起的。当枯草芽孢杆菌的蛋白质分泌最适时,在指数生长结束时和在稳定期进行对照和高产菌株的分析。显示出增加表达的基因中有htrA和htrB,它们是CssRS regulon的一部分,对高水平的蛋白质分泌和热应激作出反应。在相同的分泌压力条件下,与野生型相比,cssS突变体的转录组图谱分析显示,一些基因以CssRS依赖性方式改变了转录,例如citM,ylxF,yloA,ykoJ和flgB操纵子。但是,仅对htrA,htrB和citM观察到高亲和力的CssR结合。此外,DNA宏阵列方法揭示了孢子形成途径的几个基因被AmyQ过表达下调,并且一组运动特异性(依赖sigmaD的)转录物明显上调。随后的流式细胞仪分析表明,当AmyQ以及α-淀粉酶的不可分泌变体过量生产时,孢子形成过程将受到严重抑制。进行了类似的实验,以研究hag启动子的表达水平,hag启动子是sigmaD依赖性基因表达的公认报告子。这种方法证实了基于我们的DNA宏阵列分析的观察结果,并导致我们得出结论,在所有α-淀粉酶过量生产的条件下,与运动相关的几个基因的表达水平均维持在较高水平。
  • 【连续左旋肉碱生产过程中转基因大肠杆菌菌株的质粒维持和生理学。】 复制标题 收藏 收藏
    DOI:10.1007/s10529-007-9432-4 复制DOI
    作者列表:Bernal V,González-Veracruz M,Cánovas M,Iborra JL
    BACKGROUND & AIMS: :The effect of immobilization on cell physiology and how this determines cell metabolic performance is an important concern for developing bioprocess. This is particularly true for genetically modified microorganisms and their genetic stability. For this reason the stability and physiological state of plasmid-bearing E. coli cells were ascertained by flow cytometry. Differences in the cellular DNA and protein content (15-20%) permit discrimination of control and plasmid-bearing cells, as well as adaptation to continuous cultivation conditions in both freely suspended and immobilized states to be monitored. Moreover, the observed metabolic burden due to maintenance and over-expression of plasmid-coded genetic material and slow cell growth in poorly-viable immobilized cells were found to be the main factors contributing to strain stabilization.
    背景与目标: :固定化对细胞生理的影响以及这如何决定细胞代谢性能是发展生物过程的重要考虑因素。对于转基因微生物及其遗传稳定性尤其如此。因此,通过流式细胞术确定了带有质粒的大肠杆菌细胞的稳定性和生理状态。细胞DNA和蛋白质含量的差异(15-20%)可以区分对照细胞和带有质粒的细胞,并可以监测自由悬浮和固定状态下连续培养条件的适应性。此外,已发现由于质粒编码的遗传物质的维持和过表达以及在存活率低的固定化细胞中缓慢的细胞生长而导致的观察到的代谢负担是促成菌株稳定的主要因素。
  • 【如先前在大鼠中所证实的,成熟会降低小鼠中乙醇的最低肺泡麻醉剂浓度:没有物种差异。】 复制标题 收藏 收藏
    DOI:10.1097/00000539-199707000-00029 复制DOI
    作者列表:Fang Z,Ionescu P,Gong D,Kendig J,Harris A,Eger EI 2nd
    BACKGROUND & AIMS: The potency of conventional inhaled anesthetics increases with maturationthe 50% effective dose (minimum alveolar anesthetic concentration [MAC]) for conventional inhaled anesthetics in the neonatal rat or human exceeds MAC in the young adult. This increase also applies to ethanol in rats tested using MAC as the measure of anesthesia. However, the converse appears to be true for studies in mice assessed with the righting reflex; that is, adult mice are six times more resistant than neonates to the effects of ethanol. These disparate findings imply that maturation in rats and mice may produce opposing changes in the quantity or sensitivity of one or more receptors that mediate the actions of anesthetics that lead to the anesthetic state. Such a finding would be important for two reasons. First, both rodents are widely used in studies of anesthetic effects, and, thus, a species-dependent divergence in anesthetic effects has immediate experimental implications. Second, confirmation of such a species difference would supply an opportunity to test which receptors might be crucial to anesthetic mechanisms. Accordingly, we investigated whether maturation decreased ethanol potency in mice, using MAC as the measure of anesthesia. Applying standard techniques, we tested MAC for ethanol in 15 CF-1 mice aged 10 days (6-8.5 g) and in 13 mice aged 77-84 days (34-39 g). MAC decreased with maturation, and the decrease was indistinguishable from that found in our previous studies of rats.

    背景与目标: 常规吸入麻醉剂的效力随着成熟而增加,新生大鼠或人中常规吸入麻醉剂的50%有效剂量(最小肺泡麻醉剂浓度[MAC])超过了年轻成年人的MAC。这种增加也适用于使用MAC作为麻醉手段测试的大鼠中的乙醇。但是,对于用扶正反射评估的小鼠研究似乎相反。也就是说,成年小鼠对乙醇的抵抗力是新生小鼠的六倍。这些不同的发现暗示,大鼠和小鼠的成熟可能在介导导致麻醉状态的麻醉剂作用的一种或多种受体的数量或敏感性方面产生相反的变化。这样的发现很重要,原因有两个。首先,两种啮齿动物都广泛用于麻醉作用的研究,因此,麻醉作用中物种依赖性的差异具有直接的实验意义。其次,确认这种物种差异将提供一个机会来测试哪些受体可能对麻醉机制至关重要。因此,我们使用MAC作为麻醉手段,研究了成熟是否降低了小鼠的乙醇效力。应用标准技术,我们在10天(6-8.5 g)的15只CF-1小鼠和77-84天(34-39 g)的13只小鼠中测试了MAC的乙醇含量。 MAC随着成熟而降低,并且与我们先前的大鼠研究没有明显的区别。

  • 【研究一种标准的乞力士成分混合粉的生产,理化和感官特性。】 复制标题 收藏 收藏
    DOI:10.3109/09637489709012589 复制DOI
    作者列表:Badau MH,Igene JO,Collison EK,Nkama I
    BACKGROUND & AIMS: :The quality of kilishi, a sun dried roasted meat product vary considerably due to lack of consistent standardized non meat ingredient portion (spices and condiments) used for its preparation. In this study, an instant standard kilishi ingredient mix powder was developed. The mix powder contained 9.1% moisture, 49.7% protein, 8.3% fat, 3.2% fibre, and 4.4% ash. Particle size of the mix developed was finer than the traditional one. Yield of kilishi increased with increasing concentration of the standard mix powder used for infusion. The highest yield (87%) was obtained at 60% slurry concentration of the mix compared to 59% yield when the traditional paste was used for infusion. Sensory attributes of kilishi produced using 60% slurry concentration of the standard mix was rated better than a commercial kilishi product (P < 0.05).
    背景与目标: :由于缺乏一致的标准化非肉类成分(香料和调味品)用于制备,因此,晒干的烤肉产品kilishi的质量差异很大。在这项研究中,开发了一种即时标准的乞力士成分混合粉。混合粉末含有9.1%的水分,49.7%的蛋白质,8.3%的脂肪,3.2%的纤维和4.4%的灰分。所开发混合物的粒度比传统混合物要细。随着用于输注的标准混合粉浓度的增加,乞力士的产量增加。当混合物的浆液浓度为60%时,最高的收率(87%)获得了回报,而传统糊剂用于灌输时的收率为59%。使用60%的标准混合物浆液浓度产生的乞力士的感官属性等级优于商业乞力士产品(P <0.05)。
  • 【挥发性臭味物质与阿维链霉菌产生的阿维菌素之间的关系。】 复制标题 收藏 收藏
    DOI:10.1007/BF02932133 复制DOI
    作者列表:Rezanka T,Sobotka M,Prell A,Sigler K
    BACKGROUND & AIMS: :The time course of production of odorous compounds, i.e. geosmin and oxolones, and of avermectins was determined during the cultivation of S. avermitilis in flasks, 1- and 50-L fermentors. The amount of the antibiotics increased with increasing cultivation time up to more than 2 g/L while the concentration of geosmin rose to more than 4 mg/L. Cultivation without reflux condenser resulted in a lower product formation due to the higher stripping of geosmin. A relatively tight correlation was found between the production of geosmin and the production of avermectins. The production of oxolones peaked on cultivation days 3-5, the sum of oxolones being 60 microg/L. Subsequently, the production dropped below a measurable level. This can be explained as being due to the inhibition of oxolone production by geosmin.
    背景与目标: :在烧瓶,1升和50升发酵罐中培养阿维链霉菌的过程中,测定了产生臭味化合物即土臭素和恶臭酮以及阿维菌素的时间过程。随着培养时间的增加,抗生素的量增加到2 g / L以上,而土臭素的浓度上升到4 mg / L以上。在没有回流冷凝器的情况下进行耕种,由于土工蛋白的汽提量较高,因此产品生成量较低。在土臭素的生产与阿维菌素的生产之间发现了相对紧密的相关性。 oxolones的产生在栽培第3-5天达到顶峰,oxolones的总和为60 microg / L。随后,产量下降到可测量的水平以下。可以解释为这是由于土臭素抑制了羟考酮的产生。
  • 【瘦素的生理浓度可增加非永生化人肝星状细胞产生的胶原蛋白。】 复制标题 收藏 收藏
    DOI:10.1016/j.metabol.2006.05.016 复制DOI
    作者列表:Choudhury J,Mirshahi F,Murthy KS,Yager DR,Sanyal AJ
    BACKGROUND & AIMS: :The effects of leptin, in concentrations seen in obesity, on collagen production and turnover in non-immortalized human hepatic stellate cell (HSC), were unknown. The profibrogenic effects of leptin in these cells were studied. Hepatic stellate cells were obtained from resected livers. Collagen I/III gene expression and protein production were measured by quantitative real-time polymerase chain reaction and sodium dodecyl sulfate-polyacrylamide gel electrophoresis, respectively. The signal transduction pathways involved were evaluated by specific blockers of the phosphatidylinositol 3-kinase (PI3K), mitogen-activated protein kinase kinase (MEK), and Janus kinase 2 (JAK2). The effects on matrix metalloproteinase 1 (MMP-1) and tissue inhibitor of metalloproteinase 1 (TIMP-1) were assessed by their gene transcript levels, collagenolytic activity of cell culture supernatants, and MMP-1 protein levels. At concentrations seen in nonobese individuals ([leptin] < 10 ng/mL), leptin did not affect collagen production. At concentrations seen in obesity (30-50 ng/mL), leptin increased collagen I and III messenger RNA (mRNA) transcript levels by 286% +/- 55% (P < .001) and 167% +/- 62% (P < .007) and protein production by 45.8% +/- .02% and 84.39% +/- .01%, respectively. These effects were blocked by JAK2 inhibition as well as PI3K inhibition. Although MEK inhibition blocked leptin-induced procollagen I and III mRNA levels, there were no significant effects on collagen I and III protein levels. Leptin (10-50 ng/mL) had no significant effects on MMP-1 or TIMP-1 mRNA levels, collagenolytic activity, or MMP-1 protein levels. In conclusion, leptin, at levels seen in obese individuals, produces an increase in collagen production by HSC acting through the JAK and PI3K pathways. At these concentrations, leptin does not affect MMP-1 or TIMP-1 expression or collagenolytic activity of HSC.
    背景与目标: :瘦素在肥胖症中的浓度对非永生化人肝星状细胞(HSC)中胶原蛋白产生和更新的影响尚不清楚。研究了瘦素在这些细胞中的促纤维化作用。肝星状细胞获自切除的肝脏。通过定量实时聚合酶链反应和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分别测量胶原蛋白I / III基因的表达和蛋白质的产生。通过磷脂酰肌醇3-激酶(PI3K),有丝分裂原激活的蛋白激酶激酶(MEK)和Janus激酶2(JAK2)的特异性阻断剂评估了涉及的信号转导途径。通过基因转录本水平,细胞培养上清液的胶原蛋白水解活性和MMP-1蛋白水平评估对基质金属蛋白酶1(MMP-1)和组织金属蛋白酶1抑制剂(TIMP-1)的影响。在非肥胖个体中观察到的浓度([leptin] <10 ng / mL),leptin不会影响胶原蛋白的产生。在肥胖症中观察到的浓度(30-50 ng / mL)中,瘦素可使I型和III型胶原信使RNA(mRNA)转录水平增加286%/-55%(P <.001)和167%/-62%(P < .007)和蛋白质产量分别降低了45.8%/-.02%和84.39%/-.01%。这些作用被JAK2抑制和PI3K抑制所阻断。尽管MEK抑制阻止了瘦素诱导的胶原蛋白I和III的mRNA水平,但对胶原蛋白I和III的蛋白水平没有显着影响。瘦素(10-50 ng / mL)对MMP-1或TIMP-1 mRNA水平,胶原蛋白水解活性或MMP-1蛋白水平无明显影响。总之,在肥胖个体中观察到的瘦素水平通过HSC通过JAK和PI3K途径起作用而使胶原蛋白的产生增加。在这些浓度下,瘦素不会影响HSC的MMP-1或TIMP-1表达或胶原分解活性。
  • 【HAC15人肾上腺皮质细胞系中血管紧张素II和III的代谢及其对类固醇产生的影响。】 复制标题 收藏 收藏
    DOI:10.1210/en.2012-1557 复制DOI
    作者列表:Oki K,Kopf PG,Campbell WB,Luis Lam M,Yamazaki T,Gomez-Sanchez CE,Gomez-Sanchez EP
    BACKGROUND & AIMS: :Aldosterone is synthesized in the zona glomerulosa of the adrenal cortex under primary regulation by the renin-angiotensin system. Angiotensin II (A-II) acts through the angiotensin types 1 and 2 receptors (AT1R and AT2R). A-II is metabolized in different tissues by various enzymes to generate two heptapeptides A-III and angiotensin 1-7, which can then be catabolized into smaller peptides. A-II was more potent than A-III in stimulating aldosterone secretion in the adrenocortical cell line HAC15, and A-II, but not A-III, stimulated cortisol secretion. A-II stimulated mRNA expression of steroidogenic acute regulatory protein, 3β-hydroxysteroid dehydrogenase, CYP11B1, and CYP11B2, whereas A-III stimulated 3β-hydroxysteroid dehydrogenase, CYP11B1, and CYP11B2 but decreased the expression of CYP17A1 required for cortisol synthesis. The stimulation of aldosterone secretion by A-II and A-III was blocked by the AT1R receptor blocker, losartan, but not by an AT2R blocker. A-II was rapidly metabolized by the HAC15 cells to mainly to angiotensin 1-7, but not to A-III, and disappeared from the supernatant within 6 h. A-III was metabolized rapidly and disappeared within 1 h. In conclusion, A-II was not converted to A-III in the HAC15 cell and is the more potent stimulator of aldosterone secretion and cortisol of the two. A-III stimulated aldosterone secretion but not cortisol secretion.
    背景与目标: :醛固酮是在肾素-血管紧张素系统的主要调控下在肾上腺皮质的肾小球中合成的。血管紧张素II(A-II)通过1型和2型血管紧张素受体(AT1R和AT2R)起作用。 A-II通过各种酶在不同的组织中代谢,生成两个七肽A-III和血管紧张素1-7,然后可以将其分解为较小的肽。在刺激肾上腺皮质细胞系HAC15中的醛固酮分泌方面,A-II比A-III更有效,而A-II而不是A-III刺激了皮质醇的分泌。 A-II刺激类固醇生成的急性调节蛋白,3β-羟基类固醇脱氢酶,CYP11B1和CYP11B2的mRNA表达,而A-III刺激3β-羟基类固醇脱氢酶,CYP11B1和CYP11B2但降低了皮质醇合成所需的CYP17A1的表达。 A-II和A-III对醛固酮分泌的刺激被AT1R受体阻滞剂氯沙坦阻断,但未被AT2R阻滞剂阻断。 A-II被HAC15细胞快速代谢,主要代谢为血管紧张素1-7,但不代谢为A-III,并在6小时内从上清液中消失。 A-III快速代谢并在1​​小时内消失。总之,A-II在HAC15细胞中并未转化为A-III,而是对两者的醛固酮分泌和皮质醇更有效的刺激剂。 A-III刺激醛固酮分泌,但不刺激皮质醇分泌。

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