Break on through to the other side-biophysics and cell biology shed light on cell-penetrating peptides

Cell-penetrating peptides (CPPs) have become widely used vectors for the cellular import of molecules in basic and applied biomedical research. Despite the broad acceptance of these molecules as molecular carriers, the details of the mode of cellular internalization and membrane permeation remain elusive. Within the last two years endocytosis has been demonstrated to be a route of uptake shared by several CPPs. These findings had a significant impact on CPP research. State-of-the-art cell biology is now required to advance the understanding of the intracellular fate of the CPP and cargo molecules. Owing to their presumed ability to cross lipid bilayers, CPPs also represent highly interesting objects of biophysical research. Numerous studies have investigated structure-activity relationships of CPPs with respect to their ability to bind to a lipid bilayer or to cross this barrier. Endocytosis route only relocates the membrane permeation from the cell surface to endocytic compartments. Therefore, biophysical experiments are key to a mechanistic molecular understanding of the cellular uptake of CPPs. However, biophysical investigations have to consider the molecular environment encountered by a peptide inside and outside a cell. In this contribution we will review biophysical and cell-biology data obtained for several prominent CPPs. Furthermore, we will summarize recent findings on the cell-penetrating characteristics of antimicrobial peptides and the antimicrobial properties of CPPs. Peptides of both groups have overlapping characteristics. Therefore, both fields may greatly benefit from each other. The review will conclude with a perspective of how biophysics and cell biology may synergize even more efficiently in the future.     

Effect of antioxidant (alpha-tocopherol and ascorbic acid) fortification on light-induced flavor of milk

The effectiveness of added antioxidants against oxidation off-flavor development in light-exposed milk was evaluated using sensory and chemical analysis. Sensory testing for similarity showed no perceivable difference between control milk and milk with added (1) 0.05% alpha-tocopherol (TOC) and (2) 0.025% alpha-tocopherol and 0.025% ascorbic acid (TOC/ASC), but did demonstrate a perceivable difference when adding (3) 0.05% ascorbic acid (ASC) alone. Subsequently, sensory testing for difference showed a significant difference in oxidation off-flavor between light-exposed control milk and light-exposed milk with added TOC/ASC, whereas milk fortified with TOC was not different from control. Gas chromatography-olfactometry showed that more aroma-active flavor compounds were observed in light-exposed milk treated with TOC and TOC/ASC than light-exposed milk with no added antioxidants. The thiobarbituric acid reactive substances (TBARS) test verified chemically the extent of oxidation in control and antioxidant-treated milk samples. Milk that was exposed to light for 10 h showed a significantly higher TBARS value (0.92 +/- 0.09 mg/kg) than milk that was protected from light (0.59 +/- 0.184 mg/kg), or milk that was treated with TOC/ASC (0.26 +/- 0.092 mg/kg). Direct addition of low levels of antioxidants (TOC/ASC) to milk protected its flavor over 10 h of light exposed storage.     

Aroma analysis of light-exposed milk stored with and without natural and synthetic antioxidants

The effect of antioxidants, added in a single initial dose or in weekly additions to extended shelf life milk, was evaluated over 6 wk of lighted storage at 4 degrees C. Light-induced oxidation was measured by determining pentanal, hexanal, heptanal, and 1-octen-3-ol contents. Weekly addition of a combination of butylated hydroxyanisole and butylated hydroxytoluene (100 mg/kg of milk fat, each) maintained heptanal content of milk at levels comparable to light-protected milk, whereas an initial single addition of alpha-tocopherol significantly decreased hexanal content over the first 4 wk of storage. Odor-active compounds associated with light-induced oxidation included 2,3-butanedione, pentanal, dimethyl disulfide, hexanal, 1-hexanol, heptanal, 1-heptanol, and nonanal. The addition of butylated hydroxyanisole and butylated hydroxytoluene in a single initial addition resulted in decreases in pentanal and hexanal odor, but not in heptanal and 1-heptanol odor, whereas the addition of alpha-tocopherol and ascorbyl palmitate decreased pentanal and heptanol odor, but not hexanal and heptanal odor.     

Isolation and structure determination of cassumunarins A,B, and C: New anti-inflammatory antioxidants from a tropical ginger,Zingiber cassumunar

New antioxidants, cassumunarins A, B, and C, were isolated from the rhizomes ofZingiber cassumunar, and the structures were determined by spectroscopic methods to be complex curcuminoids. The antixidant efficiency of cassumunarins was determined by inhibitory activity of autoxidation of linoleic acid in a buffer-ethanol system. The anti-inflammatory effect was measured by inhibition of edema formation on mouse ear induced by 12-O-tetradecanoylphorbol-13-acetate. Both types of activities were stronger for the cassumunarins than for the curcumin. Visiting scholar at University of Texas (October 1992–September 1993).     

做青程度对信阳茶品质及其抗氧化活性的影响

该研究采用一芽三四叶的鲜叶原料,比较了信阳绿茶、信阳红茶和3个不同做青程度的信阳茶的品质及其抗氧化活性,以期为江北茶区小叶种茶树品种的鲜叶进一步开发利用提供理论依据。通过分光光度计、液相色谱和气相色谱-质谱联用仪检测5种茶主要生化成分和挥发性成分,利用1,1-二苯基-2-三硝基苯肼（1,1-Diphenyl-2-Picrylhydrazyl,DPPH）法检测此5种茶的抗氧化活性,最后进行感官审评。结果发现：不同做青程度的信阳茶中所含主要成分的含量及组成存在差异,随着做青程度加深,茶多酚由21.17%降到18.90%（P<0.05）,氨基酸由3.43%增加到3.73%（P<0.05）,黄酮由5.63 mg/g增加到6.55 mg/g（P<0.05）,香气成分呈递增的趋势,尤其单帖类物质增加明显,如β-蒎烯由0.13 μg/g增加到0.37 μg/g、E-β-罗勒烯0.05 μg/g增加到0.15 μg/g（P<0.05）。感官审评发现,轻度做青处理所制的茶叶品质相对较优。通过计算不同做青处理所制茶叶清除DPPH自由基半抑制浓度IC50值（Half Maximal Inhibitory Concentration,IC50）,发现随着做青程度加深,IC50值在增大,1/IC50值与茶多酚含量呈极显著正相关（P<0.01）,与氨基酸和黄酮含量呈极显著负相关（P<0.01）,与19种挥发性成分呈显著负相关（P<0.05）,说明适当的做青工艺可以改善一芽三四叶所制信阳绿茶的品质。     

毕节地区刺梨多糖提取工艺优化及产地间多糖含量与抗氧化活性差异研究

目的 优化超声辅助水提醇沉法提取刺梨多糖的工艺条件, 探究不同产地与品种间的刺梨多糖含量及抗氧化活性差异。方法 以提取率为指标, 在单因素试验基础上, 通过正交试验对超声辅助水提醇沉法工艺条件进行优化; 并以该工艺对毕节地区不同产地与品种的刺梨样品进行多糖含量的测定与分析, 对1,1-二苯基-2-苦基肼(1,1-diphenyl-2-picrylhydrazyl, DPPH)自由基和2,2’-联氨-双(3-乙基苯并噻唑啉-6-磺酸)二胺盐[2,2’-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt, ABTS]阳离子自由基清除能力进行测定, 比较各样品多糖的抗氧化活性。结果 超声辅助水提醇沉法提取刺梨多糖的最优工艺为: 提取温度75℃, 提取时间2.5 h, 料液比1:60 (g/mL); 其最佳提取率为11.88%。对37个产地的3个品种间多糖含量分析可知, 野生品种的多糖含量整体更高, 为40.87%, 纳雍县的多糖含量整体最高, 为41.40%, 刺梨多糖含量与海拔高度间未见规律性变化。各刺梨多糖对DPPH·的半抑制浓度(half maximal inhibitory concentration, IC50)明显小于对ABTS+·的IC50, 抗氧化活性未见与海拔高度存在直接相关。结论 优化后的超声辅助水提醇沉提取工艺高效可行, 各产地间多种刺梨品种多糖含量的高低呈现出一定的品种倾向性与区域偏向性, 其多糖抗氧化活性也存在较大的差异, 这可为刺梨在地区间的精准栽培及选种育种提供参考, 为刺梨资源在功能食品开发等方面提供技术和理论支持。     

榛子肽的体外抗氧化稳定性分析

利用中性蛋白酶制备榛子肽（<3 ku）,探讨环境因素对榛子肽抗氧化活性的影响,旨在为榛子肽的生产、贮藏和应用提供参考依据。随温度的升高,榛子肽的抗氧化活性显著降低（P<0.05）,在100 ℃处理3 h时,DPPH•、•OH清除能力活性维持率分别为83.61%、76.55%;pH值为6~8时榛子肽的活性可处于较高水平;当NaCl质量浓度在10 g/100 mL时,活性维持率分别为106.33%和100.25%;蔗糖浓度在2~10 g/100 mL的范围内活性维持率达到92%以上;葡萄糖质量浓度为10 g/100 mL时,活性维持率分别达到102.88%和103.39%;柠檬酸质量浓度为0.20 g/100 mL时,维持率分别为89.76%、102.64%;防腐剂在0.20 g/100 mL质量浓度,榛子肽活性能维持原有活性的80%以上;反复融冻10次,榛子肽的活性维持率分别为95.86%、95.15%;榛子肽在K+、Ca2+离子环境中的活性维持率较高;榛子肽经胃蛋白酶处理后的活性高于胃-胰蛋白酶处理。综合分析,为保持榛子肽较好的抗氧化活性,应避免长时间高温、强酸强碱及Cu2+、Zn2+离子接触,可添加适宜浓度NaCl、蔗糖、葡萄糖、柠檬酸、防腐剂,可冻融,对胃、胰蛋白酶具有耐受性。     

金枪鱼蛋白肽产品的特征性成分及其抗疲劳活性

该研究以金枪鱼碎肉为原料制备蛋白肽,并对其特征性成分、体外抗氧化活性及体内抗疲劳活性进行分析。结果表明：金枪鱼蛋白肽蛋白质含量90.11%,钙含量213 mg/100 g。呈味核苷酸及电子舌结果表明金枪鱼蛋白肽主要滋味为鲜味,且GMP对金枪鱼蛋白肽的鲜味贡献较大。电子鼻结果表明其风味成分主要为氮氧化合物、无机硫化物、甲烷等短链烷烃类化合物、有机硫化物、醇醚醛酮类化合物。体外抗氧化结果显示金枪鱼蛋白肽ABTS+和DPPH自由基清除的IC50分别为0.52 mg/mL和4.32 mg/mL,表明金枪鱼蛋白肽具有良好的体外抗氧化活性。抗疲劳结果显示中剂量组金枪鱼蛋白肽可以显著延长小鼠负重游泳时间（P<0.01）,显著降低血清尿素氮（P<0.05）和乳酸（P<0.01）含量,表明金枪鱼蛋白肽具有良好的体内抗疲劳活性。该研究利用金枪鱼碎肉制备具有独特风味和抗氧化、抗疲劳活性的优质金枪鱼蛋白肽,为金枪鱼碎肉高值化利用提供研究基础。