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1.
Duchenne muscular dystrophy (DMD) is a lethal X-inherited disease caused by dystrophin deficiency. Besides the relatively well characterized skeletal muscle degenerative processes, DMD is also associated with a dilated cardiomyopathy that leads to progressive heart failure at the end of the second decade. The aim of the present study was to characterize the diastolic Ca2+ concentration ([Ca2+]d) and diastolic Na+ concentration ([Na+]d) abnormalities in cardiomyocytes isolated from 3-, 6-, 9-, and 12-month old mdx mice using ion-selective microelectrodes. In addition, the contributions of gadolinium (Gd3+)-sensitive Ca2+ entry and inositol triphosphate (IP3) signaling pathways in abnormal [Ca2+]d and [Na+]d were investigated. Our results showed an age-dependent increase in both [Ca2+]d and [Na+]d in dystrophic cardiomyocytes compared to those isolated from age-matched wt mice. Gd3+ treatment significantly reduced both [Ca2+]d and [Na+]d at all ages. In addition, blockade of the IP3-pathway with either U-73122 or xestospongin C significantly reduced ion concentrations in dystrophic cardiomyocytes. Co-treatment with U-73122 and Gd3+ normalized both [Ca2+]d and [Na+]d at all ages in dystrophic cardiomyocytes. These data showed that loss of dystrophin in mdx cardiomyocytes produced an age-dependent intracellular Ca2+ and Na+ overload mediated at least in part by enhanced Ca2+ entry through Gd3+ sensitive transient receptor potential channels (TRPC), and by IP3 receptors.  相似文献   
2.
We have used time-resolved fluorescence resonance energy transfer (TR-FRET) to characterize the interaction between phospholamban (PLB) and the sarcoplasmic reticulum (SR) Ca-ATPase (SERCA) under conditions that relieve SERCA inhibition. Unphosphorylated PLB inhibits SERCA in cardiac SR, but inhibition is relieved by either micromolar Ca2+ or PLB phosphorylation. In both cases, it has been proposed that inhibition is relieved by dissociation of the complex. To test this hypothesis, we attached fluorophores to the cytoplasmic domains of SERCA and PLB, and reconstituted them functionally in lipid bilayers. TR-FRET, which permitted simultaneous measurement of SERCA–PLB binding and structure, was measured as a function of PLB phosphorylation and [Ca2+]. In all cases, two structural states of the SERCA–PLB complex were resolved, probably corresponding to the previously described T and R structural states of the PLB cytoplasmic domain. Phosphorylation of PLB at S16 completely relieved inhibition, partially dissociated the SERCA–PLB complex, and shifted the T/R equilibrium within the bound complex toward the R state. Since the PLB concentration in cardiac SR is at least 10 times that in our FRET measurements, we calculate that most of SERCA contains bound phosphorylated PLB in cardiac SR, even after complete phosphorylation. 4 μM Ca2+ completely relieved inhibition but did not induce a detectable change in SERCA–PLB binding or cytoplasmic domain structure, suggesting a mechanism involving structural changes in SERCA’s transmembrane domain. We conclude that Ca2+ and PLB phosphorylation relieve SERCA–PLB inhibition by distinct mechanisms, but both are achieved primarily by structural changes within the SERCA–PLB complex, not by dissociation of that complex.  相似文献   
3.
Novel synthetic mimics of antimicrobial peptides have been developed to exhibit structural properties and antimicrobial activity similar to those of natural antimicrobial peptides (AMPs) of the innate immune system. These molecules have a number of potential advantages over conventional antibiotics, including reduced bacterial resistance, cost-effective preparation, and customizable designs. In this study, we investigate a family of nylon-3 polymer-based antimicrobials. By combining vesicle dye leakage, bacterial permeation, and bactericidal assays with small-angle X-ray scattering (SAXS), we find that these polymers are capable of two interdependent mechanisms of action: permeation of bacterial membranes and binding to intracellular targets such as DNA, with the latter necessarily dependent on the former. We systemically examine polymer-induced membrane deformation modes across a range of lipid compositions that mimic both bacteria and mammalian cell membranes. The results show that the polymers' ability to generate negative Gaussian curvature (NGC), a topological requirement for membrane permeation and cellular entry, in model Escherichia coli membranes correlates with their ability to permeate membranes without complete membrane disruption and kill E. coli cells. Our findings suggest that these polymers operate with a concentration-dependent mechanism of action: at low concentrations permeation and DNA binding occur without membrane disruption, while at high concentrations complete disruption of the membrane occurs. This article is part of a Special Issue entitled: Interfacially Active Peptides and Proteins. Guest Editors: William C. Wimley and Kalina Hristova.  相似文献   
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【目的】测定黄粉虫Tenebrio molitor Linnaeus幼虫抗菌肽提取液的浓度、抑菌活性及其部分生化特性和凝血效应。【方法】本实验用浓度为1×108 CFU/m L大肠杆菌Escherichia coli诱导5龄黄粉虫幼虫,分别在诱导12、24、36、48、60和72 h后提取其中的抗菌肽,并用考马斯亮兰法测定抗菌肽粗提液蛋白的浓度,并用滤纸片法测定其抑菌活性,同时对其热稳定性、反复冻融稳定性、蛋白酶稳定性及不同p H对其活性的影响等生化特性及凝血效应进行了探究。【结果】经大肠杆菌诱导的黄粉虫抗菌肽粗提液的蛋白浓度均显著高于未诱导的黄粉虫组(P<0.01),且在诱导48 h时产生的抗菌肽提取液蛋白的浓度最高,产生的抑菌圈直径也显著高于未诱导的黄粉虫组(P<0.05),生化特性的测定结果显示,黄粉虫抗菌肽有较好的热稳定性、酶稳定性及酸碱稳定性,反复冻溶后对其抑菌活性影响不大,并且无凝血效应。【结论】大肠杆菌可以刺激黄粉虫的免疫系统,增加抗菌肽的表达量,使其产生浓度高、活性强的抗菌肽,且生化特性较稳定。本研究对黄粉虫抗菌肽作为绿色抗生素用于畜牧养殖业的进一步开发与利用提供了科学的理论依据。  相似文献   
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7.
Subcellular volumes and metabolite concentrations in barley leaves   总被引:26,自引:1,他引:25  
Metabolite concentrations in subcellular compartments from mature barley (Hordeum vulgare L. cv. Apex) leaves after 9 h of illumination and 5 h of darkness were determined by nonaqueous fractionation and by the stereological evaluation of cellular and subcellular volumes from light and electron micrographs. Twenty one-day-old primary leaves of barley with a total leaf volume of 902 μL per mg chlorophyll were found to be composed of 27% epidermis, 42% mesophyll cells, 6% veins, 4.5% apoplast and 23% gas space. While in epidermal cells 99% of the volume was occupied by the vacuole, mesophyll cells with an average volume of 31.3 pL consisted of 23 pL (73%) vacuole, 4.6 pL (19%) chloroplasts, 2.06 pL (6,7%) cytosol (including smaller organelles and vesicles), 0.34 pL (1%) mitochondria and 107 fL (0.34%) nucleus. The differences between leaves harvested after 9 h of illumination and after 5 h of darkness were in the size of the stromal compartment and the starch grains therein. Subcellular metabolite concentrations were calculated from the compartmental volumes and metabolite contents of the compartments as determined by nonaqueous fractionation. The amino-acid concentrations in stroma and cytosol were rather similar after 9 h of illumination and 5 h of darkness. In contrast, the vacuolar amino-acid concentrations were about one order of magnitude lower than the stroma and cytosol values, and there was a slight increase in concentration after 5 h of darkness.  相似文献   
8.
An investigation on the influence of lead toxicity on some of the hepatic enzymes was studied in rats both after a shorter interval of 15 d and after longer intervals of 60 and 90 d. Three different doses of lead as 5, 10, and 50 mg/kg body wt were administered orally on every alternate day. Whereas significant inhibition of succinic dehydrogenase was seen following lead poisoning, the activity acid and alkaline phosphatase increased with lead intoxication. The histoarchitecture of the liver was grossly intact. Liver accumulated less lead compared to kidney at 60 and 90 d.  相似文献   
9.
Wind-induced sediment resuspension occurs frequently in the shallow and eutrophic Lake Arresø, Denmark. The impact of resuspension on internal phosphorus loading was investigated by laboratory experiments studying P-release from the undisturbed sediment surface and by experiments simulating resuspension events.Phosphorus release from undisturbed sediment sampled in May and August was 12 mg and 4 mg m–2 d–1, respectively. During experimental simulation of resuspension, soluble reactive phosphate (SRP) increased by 20–80 µg l–1, which indicates that a typical resuspension event in the lake would be accompanied by the release of 150 mg SRP m–2. The internal P loading induced by resuspension is estimated to be 60–70 mg m–2 d–1, or 20–30 times greater than the release from undisturbed sediment.SRP release during simulation of resuspension was mainly dependent on the equilibrium conditions in the water column and was basically independent of the increase in suspended solids and the duration of resuspension. A second simulation of resuspension conducted 26 hours later, did not result in any further release of SRP from sediment sampled in May. In contrast, there was an additional SRP release from sediment sampled in August, indicating that an exchangable P pool, capable of altering equilibrium conditions, is built up between resuspension events.It is concluded that resuspension, by increasing the P flux between sediment and water, plays a major role in the maintenance of the high nutrient level in Lake Arresø. A relatively high release rate is maintained during resuspension because of the low Fe:P ratio and the high concentration of NH4Cl-extractable P in the sediment.  相似文献   
10.
水稻上部叶片叶绿素含量的高光谱估算模型   总被引:10,自引:1,他引:9  
杨杰  田永超  姚霞  曹卫星  张玉森  朱艳 《生态学报》2009,29(12):6561-6571
叶片叶绿素 (Chl) 状况是评价植株光合效率和营养胁迫的重要指标,实时无损监测Chl状况对作物生长诊断及氮素管理具有重要意义.以不同生态点、不同年份、不同施氮水平、不同类型水稻品种的4个田间试验为基础,于主要生育期同步测定了水稻主茎顶部4张叶片的高光谱反射率及Chl含量,并计算了350~2500 nm范围内任意两波段组合而成的比值(SR[λ1,λ2])和归一化(ND[λ1,λ2])光谱指数以及已报道的对Chl敏感的光谱指数,进一步系统分析了叶片Chl含量与上述光谱指数之间的定量关系.结果表明,红边波段的比值和归一化光谱指数可以较好地预测水稻上部4叶的Chl含量(R~2>0.9),但对于不同Chl指标其最佳组合波段有所差异.估算叶绿素a (Chla)、叶绿素总量(Chla+b)和叶绿素b (Chlb)的最佳比值光谱指数分别为SR(724,709)、SR(728,709)和SR(749,745),方程拟合决定系数R~2分别是0.947、0.946、0.905;最佳归一化光谱指数分别为ND(780,709)、ND(780,712)和ND(749,745),R~2分别是0.944、0.943、0.905.引入445 nm波段反射率对上述光谱指数进行修正,可以降低叶片表面反射差异的影响,提高模型的应用范围.利用不同年份独立的试验资料对所建模型进行了检验,结果表明,修正型比值光谱指数 mSR(724,709)、mSR(728,709) 和 mSR(749,745),以及修正型归一化光谱指数mND(780,709)、mND(780,712) 和 mND(749,745) 预测 Chla、Chla+b 和 Chlb 的效果更好,其测试的RMSE分别为 0.169、0.192、0.052、0.159、0.176、0.052,RE分别为8.18%、7.74%、13.01%、8.26%、7.59%、12.96%,均较修正前降低,说明修正后的光谱指数普适性更好.  相似文献   
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