Affinity and kinetic modulation of polyamide–DNA interactions by N‐modification of the heterocycles

Synthetic N‐methyl imidazole and N‐pyrrole containing polyamides (PAs) that can form “stacked” dimers can be programmed to target and bind to specific DNA sequences and control gene expression. To accomplish this goal, the development of PAs with lower molecular mass which allows for the molecules to rapidly penetrate cells and localize in the nucleus, along with increased water solubility, while maintaining DNA binding sequence specificity and high binding affinity is key. To meet these challenges, six novel f‐ImPy*Im PA derivatives that contain different orthogonally positioned moieties were designed to target 5′‐ACGCGT‐3′. The synthesis and biophysical characterization of six f‐ImPy*Im were determined by CD, ΔT_M, DNase I footprinting, SPR, and ITC studies, and were compared with those of their parent compound, f‐ImPyIm. The results gave evidence for the minor groove binding and selectivity of PAs 1 and 6 for the cognate sequence 5′‐ACGCGT‐3′, and with strong affinity, K_eq = 2.8 × 10⁸ M^?1 and K_eq = 6.2 × 10⁷ M^?1, respectively. The six novel PAs presented in this study demonstrated increased water solubility, while maintaining low molecular mass, sequence specificity, and binding affinity, addressing key issues in therapeutic development. © 2013 Wiley Periodicals, Inc. Biopolymers 99: 497–507, 2013.     

Evaluation of heat shock protein 70 as a biomarker of environmental stress in Fucus serratus and Lemna minor

Heat shock proteins (Hsps) are known to be induced in response to short-term stress. The present study aimed to evaluate the potential of Hsp70 as a biomarker of stress produced by increased temperature, osmotic pressure, and exposure to cadmium and sodium chloride in marine macroalgae and fresh water plant species. An indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) was developed with a working range of 0.025-10 μg ml^-1 using a monoclonal antibody raised against purified Hsp70 of Phaseolus aureus (mung bean). Fucus serratus (toothed wrack), Chondrus crispus (Stackhouse or Carrageen moss), Ulva lactuca (sea lettuce) and Lemna minor (common duckweed) sample extracts were stressed for up to 24 h and then tested in the IC-ELISA. The presence of Hsp70 and cross-reactivity of the monoclonal antibody was confirmed by Western blot. The heat shock response was confirmed in each species using a 2-h 42°C treatment. Following heat shock, Hsp70 concentrations increased to a peak at 2 h (F. serratus) or 4 h (L. minor), after which concentrations decreased. Osmotic and cadmium stresses also resulted in elevated Hsp70 concentrations in samples of F. serratus and L. minor when compared with unstressed controls. In both, osmotic and metal stress, the production of Hsp70 increased to a maximum and subsequently decreased as the stressor levels increased. Results suggest that Hsp70 IC-ELISA could potentially be applied to the detection of stress in these aquatic species, although it would probably be most effective when used in conjunction with other measurements to provide a stressor-specific biomarker profile or fingerprint.     

浑善达克沙地四种生境中榆树天然更新幼苗发育的比较

对浑善达克沙地腹地风蚀低地、覆沙草地、平坦流动沙地和流动沙丘阴坡 4种类型生境中植物群落和榆树幼苗生长的比较研究结果表明 :(1)尽管单个样方内植物种数生境间没有差异 ,但群落总盖度覆沙草地的最低 ,流动沙丘阴坡上的居中 ,风蚀低地和流沙平地的最高。榆树幼苗的分盖度风蚀低地的最高 ,在流沙平地的次之 ,在覆沙草地的最低 ,流动沙丘阴坡的介于流沙平地和覆沙草地的中间 ,但与二者没有显著性差异 ;(2 )风蚀低洼地的幼苗生长的最好 ,平均高度、主根长度、叶片数和叶面积均显著大于生长在覆沙草地和流沙平地的幼苗的。生长在流动沙丘阴坡上幼苗的植株高度、叶片数目和叶面积介于它们之间 ,主根长度与风蚀低地的没有显著差异 ;(3)风蚀低洼地榆树幼苗根、茎、叶各部分生物量都明显地高于生长在其它 3种生境中幼苗的 ,流动沙丘阴坡上榆树幼苗的生物量还明显高于生长在覆沙草地和流沙平地上的 ,生长在覆沙草地和流沙平地的榆树幼苗的生物量基本上没有差异。该结果说明 ,榆树幼苗在风蚀低地生长最好 ,其次是在流动沙丘阴坡 ,这两种生境可能是浑善达克沙地榆树更新的主要地方。     

不同生育期榆树净光合速率对生态因子和生理因子的响应

利用Li-6400便携式光合作用系统,观测了内蒙古浑善达克沙地榆树(Ulmus pumila)的净光合速率(Pn)、上表皮气孔导度(Gs-u)、下表皮气孔导度(Gs-l)等生理因子及大气温度(Ta)、光合有效辐射(PAR)、大气二氧化碳浓度(Ca)和空气相对湿度(RH)等生态因子,应用相关分析、通径分析、决策系数分析及灰色关联分析,系统的研究了榆树Pn在不同生育期的日变化特征及其与生态因子和生理因子间的关系,为深入揭示其生理特征及浑善达克沙地榆树规范化种植提供理论依据。研究表明:1榆树Pn日变化在各生育期均呈双峰型曲线,表现出不同程度的光合"午休"。2榆树Pn在各生育期大小顺序为8月7月9月,其值分别为8.18、7.92μmol CO2m-2s-1和6.52μmol CO2m-2s-1。37、8、9月份PAR、Gs-u与榆树Pn呈显著正相关,且PAR的直接作用和决策系数均最大,Gs-u的直接作用大于其他因子的间接作用,且决策系数仅次于PAR,其他因子在各生育期也表现出对榆树Pn不同程度的影响。4不同生育期榆树对各生态因子和生理因子的响应有所不同,7月份Ta为Pn主要限制变量,8月份RH为Pn主要限制变量,9月份各因子均起增进作用。5不同生育期榆树Pn的变化主要受PAR和Gs-u的影响,但不存在简单的线性相关关系,是所有因子综合影响的结果。     

坝下地区防护林不同树种生长特征

采用标准样地调查法对防护林林分结构指标进行调查,研究河北坝下地区丰宁县小坝子乡不同防护林树种榆树、杨树及油松的动态变化规律.结果表明: 该区域榆树、杨树和油松纯林的平均年龄均为10年,杨树纯林的平均胸径和树高分别是榆树纯林的2.3和3.8倍,是油松纯林的2.3和3.0倍.杨树纯林的平均胸径、树高、叶面积指数(LAI)、活枝下高等指标显著大于榆树纯林和油松纯林.与杨树混交林相比,杨树纯林的林分密度高10.8%,纯林平均胸径则低5.2%,平均树高低11.3%;与榆树混交林相比,榆树纯林的林分密度高6.6%,纯林平均胸径、树高分别低7.8%和14.2%;与油松混交林相比,油松纯林的林分密度大4.9%,纯林平均胸径和树高分别低29.3%和31.8%.各林分类型的平均胸径、树高与密度呈显著负相关;平均LAI与密度和活枝下高呈显著正相关,与胸径、树高呈显著负相关;平均活枝下高与密度呈显著正相关.针阔混交林的胸径和树高生长显著优于针叶纯林.该区域防护林综合生长潜力呈上升趋势,而横向生长潜力总体上呈下降趋势.     

Synthesis and anticancer activity of new thiazolo[3,2-a]pyrimidines: DNA binding and molecular modeling study

A novel series of nitrogenous heterocycles starting from chalcones including thiazolo[3,2-a]pyrimidines (20–67), were synthesized. Structure elucidation of the synthesized compounds was attained by the use of ¹H NMR, ¹³CC NMR, and Mass spectrometry. The obtained compounds were evaluated for their in vitro anticancer activity at the National Cancer Institute (NCI) 60 cell lines panel assay. Three cell lines, non-small cell lung cancer Hop-92, ovarian cancer IGROV1, and melanoma SK-MEL-2, exhibited some sensitivity against most of the tested compounds. Six compounds have passed the 5-log dose level NCI assay. Compounds 34 and 24 proved to be the most active derivatives with GI₅₀, TGI, LC₅₀ of 5.89, 20.0, 66.1% and 5.0, 19.5, 52.5% respectively. Compounds 36, 39, 63 showed lesser activity with GI₅₀, TGI, LC₅₀ 3.2, 11.8, 38.9%, 3.4, 16.6, 60.3%, 3.5, 17.8, 66.1% respectively. DNA binding assay of synthesized compound were performed. Molecular docking showed that compounds 34, 42, and 60 could effectively fit into the minor groove and selectively bind with AT base pairs. The results could confer the anticancer activity of compounds 24, 34, 36, and 39 in vitro to their abilities to bind at DNA minor groove.     

Establishment and characterization of American elm cell suspension cultures

Cell suspension cultures of Dutch elm disease (DED)-tolerant and DED-susceptible American elms clones have been established and characterized as prerequisites for contrasts of cellular responses to pathogen-derived elicitors. Characteristics of cultured elm cell growth were monitored by A₇₀₀ and media conductivity. Combined cell growth data for all experiments within a genotype showed relatively low variances and between-genotype contrasts during repeated passages showed no significant differences. Subculturing exponentially growing cells at 8–14 day intervals, within readily measured parameters of media conductivity (4.95–4.2 mmhos) and cell concentration (≥ 1.4 A₇₀₀), consistently resulted in repeatable profiles of elm cell growth and minimized lag phase. Culture cells were essentially homogeneous after 5 subculture passages and their overall appearance was stable. We conclude that the described procedure resulted in consistent cultures suitable for elicitor treatment experiments. This revised version was published online in June 2006 with corrections to the Cover Date.     

Scaling the leaf length-times-width equation to predict total leaf area of shoots

Background and AimsAn individual plant consists of different-sized shoots, each of which consists of different-sized leaves. To predict plant-level physiological responses from the responses of individual leaves, modelling this within-shoot leaf size variation is necessary. Within-plant leaf trait variation has been well investigated in canopy photosynthesis models but less so in plant allometry. Therefore, integration of these two different approaches is needed.MethodsWe focused on an established leaf-level relationship that the area of an individual leaf lamina is proportional to the product of its length and width. The geometric interpretation of this equation is that different-sized leaf laminas from a single species share the same basic form. Based on this shared basic form, we synthesized a new length-times-width equation predicting total shoot leaf area from the collective dimensions of leaves that comprise a shoot. Furthermore, we showed that several previously established empirical relationships, including the allometric relationships between total shoot leaf area, maximum individual leaf length within the shoot and total leaf number of the shoot, can be unified under the same geometric argument. We tested the model predictions using five species, all of which have simple leaves, selected from diverse taxa (Magnoliids, monocots and eudicots) and from different growth forms (trees, erect herbs and rosette herbs).Key ResultsFor all five species, the length-times-width equation explained within-species variation of total leaf area of a shoot with high accuracy (R² > 0.994). These strong relationships existed despite leaf dimensions scaling very differently between species. We also found good support for all derived predictions from the model (R² > 0.85).ConclusionsOur model can be incorporated to improve previous models of allometry that do not consider within-shoot size variation of individual leaves, providing a cross-scale linkage between individual leaf-size variation and shoot-size variation.     

放牧方式对浑善达克沙地榆树疏林退化的影响

浑善达克沙地榆树疏林是分布于草原地带的隐域植被类型,相较周边的典型草原区,其植被更加茂密,乔灌丛生,水草丰美,千百年来一直是牧民的优质冬季牧场。近半个世纪以来,因人类的过度开垦以及不合理的放牧管理,浑善达克沙地植被遭到空前的破坏,沙丘活化,载畜能力降低,生态价值和经济价值严重受损。近年来,随着国家草畜平衡以及禁牧政策的推广落实,放牧的牲畜总量得到一定程度的遏制,然而大面积草场还在继续退化。在牧民对生产生活的基本需求下,牲畜总量不可能无限制的压制。另外,适度的放牧对草原生态系统健康是有益的。因此,如何改良放牧方式,合理利用草场,在保持生态良好的基础上合理发挥草场的畜牧价值,是我们亟待探索的问题。以浑善达克沙地的典型天然植被榆树疏林为例,对不同放牧方式下的植被进行调查,基于沙地特殊的基质和植被特征,探讨适合沙地的放牧利用方式。研究表明,在相似的放牧强度下,把沙地作为冬营地,其榆树种群更新正常,植被覆盖度以及植物种类等均能保持良好,而把沙地作为夏季营地,榆树疏林植被退化严重,具体表现为:(1)榆树种群自然更新受阻;(2)灌木群落大量死亡或消失;(3)草本覆盖度显著降低,植物种类减少,多年生草...     

两种浮萍植物的叶绿体超微结构对模拟酸雨的敏感性

用蒸馏水和不同pH值(5.5,4.5,3.5和2.5)的模拟酸雨培养浮萍(Lemn aminorL.)和紫萍(Spirodela polyrrihiza(L.)Schleid)48h后,测定叶状体膜系统渗漏率,观察叶绿体超微结构的变化。结果表明,随酸雨pH的下降,两种浮萍叶状体的膜系统渗率增大,叶绿体超微结构受损。浮萍和紫萍对酸雨的敏感性和伤害时细胞与叶绿体形态变化显示一定的种间差别。浮萍的结构性损伤始于pH4.5,原生质体收缩,出现质壁分离,基粒结构混乱;pH3.5时叶绿体肿胀呈球状,片层结构破坏并出现许多空泡。紫萍在pH3.5时膜系统的外渗率仍较低,基粒和基质类囊体结构无明显改变;pH2.5时叶绿体结构才出现严重伤害,但未见明显的肿胀与质壁分离现象。因此认为在两种浮萍的共生水体中,紫萍对酸雨污染有较强的生存竞争能力,而浮萍则可用作pH<4.5水体的灵敏指示植物。