P-3A and (−)-desacetamido P-3A: demonstration and Study of their effective functional cleavage of duplex DNA

A study of the Fe(II) complexes of P-3A (1) and (−)-desacetamido P-3A (2) abilities to cleave duplex DNA was conducted through examination of single-strand and double-strand cleavage of supercoiled φX174 RFI DNA (Form I) in the presence of O₂ to produce relaxed (Form II) and linear (Form III) DNA, respectively. Like Fe(II)-bleomycin A₂ and deglycobleomycin A₂, Fe(II)-1 and 2 effectively produced both single- and double-strand cleavage of supercoiled φX174 DNA. Unlike Fe(II)-bleomycin A₂ or deglycobleomycin A₂, Fe(II)-1 and 2 were found to cleave duplex w794 DNA with no discemible sequence selectively suggesting that the polynucleotide recognition of the C-terminus tetrapeptide S subunit of the bleomycins including the bithiazole may dominate the bleomycin A₂ DNA cleavage selectivity.     

水氮组合对冬小麦干物质及氮素积累和产量的影响

于2015—2017年小麦生长季在山东省泰安市农业科学研究院肥城试验基地进行田间试验,供试材料为‘泰山28',在150(A₁)、300(A₂)、450(A₃)、600 m³·hm^-2(A₄)4个灌水量和90(B₁)、135(B₂)、180(B₃)、225 kg·hm^-2(B₄)4个施氮水平下,研究水氮组合对小麦生长发育过程中干物质积累、氮素积累、水分消耗利用、光合特性、籽粒产量等的影响。结果表明: A₃B₃条件下各生育阶段的干物质积累量和氮素积累量,成熟期籽粒干物质和氮素积累量均为最大,花前花后营养器官生产储藏干物质及氮素向籽粒的运输量最高,且与其他水氮组合处理差异显著。各氮素处理下,60~200 cm土层土壤耗水量均为A₃>A₄>A₂>A₁;A₃B₃处理下的水分利用效率和氮素利用效率高于A₃B₄、A₄B₃和A₄B₄。A₃B₃处理显著提高了开花后7~28 d的旗叶净光合速率、气孔导度和蒸腾速率,有利于小麦进行光合作用合成碳水化合物。水氮组合效应显著影响籽粒产量和产量构成,且A₃B₃处理下小麦产量最高,达到9400 kg·hm^-2。综上,450 m³·hm^-2和180 kg·hm^-2的水氮组合处理可以显著提高小麦干物质和氮素积累量,并促进干物质和氮素向籽粒运输,与高水肥处理相比,可以有效提高水分利用效率和氮素利用效率,有利于增强小麦旗叶的光合能力,产生更多的碳水化合物,增加籽粒产量。     

灌水下限及秸秆还田对温室番茄产量、品质和水分利用效率的影响

为了探究秸秆还田滴灌灌水下限和秸秆还田量对温室番茄产量、品质和水分利用效率的影响,在温室内进行裂区试验。秸秆还田时间分别为1年(2018年)、2年(2017年)和3年(2016年),设置4个秸秆还田量(0、1.5×10⁴、3×10⁴、4.5×10⁴ kg·hm^-2)和4个灌水下限(50%θ_f、60%θ_f、70%θ_f、80%θ_f,θ_f为田间持水量),对土壤含水率、番茄产量和品质进行监测。采用方差分析、熵权法和TOPSIS法对番茄产量、品质和水分利用效率进行分析。结果表明: 番茄产量随灌水下限增大而增大,在灌水下限为80%θ_f时产量最大,秸秆还田第1、2和3年,最大平均产量分别为93.55、87.23和99.34 t·hm^-2。水分利用效率和品质指标均随灌水下限的升高而降低。在秸秆还田第1年时,秸秆还田量为1.5×10⁴ kg·hm^-2时番茄平均产量达到最大值,为99.60 t·hm^-2;在秸秆还田第2、3年时,秸秆还田量为4.5×10⁴ kg·hm^-2时番茄平均产量最大,分别为92.50和107.75 t·hm^-2。番茄水分利用效率在秸秆还田第1、2年,秸秆还田量为1.5×10⁴ kg·hm^-2时达到最大;在秸秆还田第3年时,秸秆还田量为4.5×10⁴ kg·hm^-2达到最大。番茄的品质指标随秸秆还田年限和秸秆还田量增加表现出不同趋势。     

Bioconversion of phospholipids by immobilized phospholipase A2

Phospholipase A₂ selectively hydrolyses the ester linkage at the sn-2 position of phospholipids forming lysocompounds. This bioconversion has importance in biotechnology since lysophospholipids are strong bioemulsifiers. The aim of the present work was to study the kinetic behaviour and properties of immobilized phospholipase A₂ from bee venom adsorbed into an ion exchange support. The enzyme had high affinity for CM-Sephadex^® support and the non-covalent interaction was optimum at pH 8. The activity of immobilized phospholipase A₂ was comparatively evaluated with the soluble enzyme using a phospholipid/Triton X-100 mixed micelle as assay system. The immobilized enzyme showed high retention activity and excellent stability under storage. The activity of the immobilized system remained almost constant after several cycles of hydrolysis. Immobilized phospholipase A₂ was less sensitive to pH changes compared to soluble form. The kinetic parameters obtained (V_max 883.4 μmol mg⁻¹ min⁻¹ and a K_m 12.9 mM for soluble form and V_max = 306 μmol mg⁻¹ min⁻¹ and a K_m = 3.9 for immobilized phospholipase A₂) were in agreement with the immobilization effect. The results obtained with CM-Sephadex^®-phospholipase A₂ system give a good framework for the development of a continuous phospholipid bioconversion process.     

活性焦对晋北盐碱地土壤性质和两种植物生长的影响

固废基改良剂是盐碱地改良的有效手段之一。活性焦作为一种多孔废弃物,应用于盐碱地预期可以起到改善土壤性质、减缓植物盐碱胁迫的作用。为了阐明活性焦对晋北盐碱地的改良效应,本研究设置不同用量活性焦(CK, 0 g·kg^-1;A₁₀, 10 g·kg^-1;A₂₀, 20 g·kg^-1;A₅₀, 50 g·kg^-1),分析其对盐碱地土壤性质和植物生长的影响。结果表明: 施用活性焦能显著提高土壤的水稳性团聚体含量,并降低土壤盐分含量、pH值和电导率。与CK相比,种植碱茅和玉米土壤的团聚体平均重量直径增加5.1%～32.2%,pH值降低0.4%～4.1%,钠吸附比(SAR)降低4.8%～18.7%,电导率降低7.4%～8.2%。施用适量活性焦能够降低植物细胞的质膜损伤,提高叶片叶绿素和Ca²⁺含量,从而有效促进植物生长,碱茅和玉米的生物量均在A₂₀处理达到最大。表明盐碱土施用20 g·kg^-1活性焦(A₂₀)可改善根际土壤环境,提高植物对Ca²⁺等的选择性吸收,从而降低盐分对植物细胞的损伤,促进盐碱生境中植物的生长。     

Interaction of ovokinin(2-7) with vascular bradykinin 2 receptors

Intravenous administration of ovokinin(2–7), a cleavage peptide derived from ovalbumin, dose-dependently (0.1–5 mg/kg) lowered the mean arterial pressure (MAP) that was not accompanied by a significant change in the heart rate (HR) of urethane-anesthetized rats. The hypotensive effects of ovokinin(2–7) were five orders of magnitude lower compared to that of bradykinin and were largely prevented by pretreatment with the bradykinin B₂ receptor antagonist HOE140 (81.6±18.4%) and moderately affected by the B₁ receptor antagonist [des-Arg¹⁰]-HOE140 (26.3±15.5%). Intracellular Ca²⁺ levels, as measured by Fur 2-AM, were significantly elevated in cultured aorta smooth muscle cells by ovokinin(2–7). The increases were abolished by HOE140 and unaffected by [des-Arg¹⁰]-HOE140. The elevation of intracellular Ca²⁺ by ovokinin(2–7) was dependent on Ca²⁺ entry from extracellular space as it was reduced in a Ca²⁺-free solution. Pretreatment of the cells with the phospholipase C inhibitor U73122 (2 μM) eliminated the Ca²⁺ increase by the peptide. PA phosphohydrolase and phospholipase A₂ inhibitors significantly reduced the responses as well. Our results show that ovokinin(2–7) modulates cardiovascular activity by interacting with B₂ bradykinin receptors.     

高原鼠兔种群生产量生态学的研究Ⅲ.高原鼠兔生命率非密度制约与密度制约的种群动态趋势

本文以高原鼠兔(Ochotona curzoniae)自然种群生命表的统计参数为基础,根据非密度制约Leslie模型及具有密度制约反馈的标准Leslie修正模型,分别预测了该种群在1982-2001年间的发展趋势。在菲密度制约条件下,该种群呈指数增长。在密度制约存在肘,种群增长趋于平衡状态,且存滔率密度制约较繁殖率密度制约对种群的作用更大。存活率密度制约与非密度制约的年龄结构均为Leslie分布,繁殖率密度制约作用的种群稳定年龄分布更平均,其平衡状态的种群大小则由模型的参数决定。     

Involvement of phospholipase A2 in norepinephrine release from synaptosomes isolated from rat cerebral cortex

Phospholipase A₂ added directly to superfused [³H]norepinephrine-labeled synaptosomes could cause the release of neurotransmitter molecules. Chloroquine and quinacrine, which block the action of phospholipase A₂, inhibited either the phospholipase A₂-stimulated or the high potassium-stimulated release of [³H]norepinephrine from synaptosomes. Only quinacrine blocked the high potassium-stimulated influx of Ca²⁺. It appears that during stimulation of synaptosomes, Ca²⁺ influx leads to the activation of phospholipase A₂, which in turn, hydrolyzes membrane phospholipids in situ. The formation of lysophospholipids may alter the microenvironment and the physicochemical properties of membranes, resulting in the release of neurotransmitter through exocytosis.     

ESR characteristics of Photosystem I in deuterium oxide: Further evidence that electron acceptor A1 is a quinone

The appearance of ESR signals from Photosystem I (PS I) electron acceptors A₁ and A₀ in water or deuterium oxide suspension was followed using a low-temperature photoaccumulation technique. In deuterated samples the A₁ signal was narrowed by a factor of 0.66 compared with the control. This effect was fully reversible upon resuspension of treated samples in H₂O. The narrow ESR signal from deuterated A₁ had similar power saturation characteristics to the normal signal; however, a signal from a second component resolved by deuteration was saturated at higher microwave powers than the control. The power saturation behaviour of A⁻₁ in un-modified reaction centres indicated that it is an anionic semiquinone in a ‘protic’ environment. Deuteration reversibly modified the relative extents of reduction of iron sulphur electron acceptors A and B such that centre B became the more stable electron acceptor. The g-value and line-width of iron sulphur centre X was not modified by deuteration although it appeared to become more efficiently reduced. These results are discussed in the light of current evidence from optical, electron spin polarisation and extraction experiments that suggest that A₁ is a quinone, probably vitamin K-1.     

Design of (N)-methanocarba adenosine 5'-uronamides as species-independent A3 receptor-selective agonists

2-Chloro-5′-N-methylcarboxamidoadenosine analogues containing the (N)-methanocarba (bicyclo[3.1.0]hexane) ring system as a ribose substitute display increased selectivity as agonists of the human A₃ adenosine receptor (AR). However, the selectivity in mouse was greatly reduced due to an increased tolerance of this ring system at the mouse A₁AR. Therefore, we varied substituents at the N⁶ and C2 positions in search of compounds that have improved A₃AR selectivity and are species independent. An N⁶-methyl analogue was balanced in affinity at mouse A₁/A₃ARs, with high selectivity in comparison to the A_2AAR. Substitution of the 2-chloro atom with larger and more hydrophobic substituents, such as iodo and alkynyl groups, tended to increase the A₃AR selectivity (up to 430-fold) in mouse and preserve it in human. Extended and chemically functionalized alkynyl chains attached at the C2 position of the purine moiety preserved A₃AR selectivity more effectively than similar chains attached at the 3-position of the N⁶-benzyl group.     

短期CO2浓度升高和干旱胁迫对白羊草土壤碳氮和微生物根际效应的影响

采用人工气候室和盆栽控水试验研究黄土丘陵区典型草本植物白羊草在倍增CO₂浓度(800 μmol·mol^-1)下和充分供水(75%～80%的田间持水量)、轻度干旱胁迫(55%～60%的田间持水量)和重度干旱胁迫(35%～40%的田间持水量)下根际和非根际土壤碳氮含量和微生物群落结构及其根际效应.结果表明: CO₂浓度升高和干旱胁迫对白羊草根际和非根际土壤有机碳、全氮和水溶性有机碳(DOC)含量及其根际效应均无显著影响.轻度干旱胁迫下CO₂浓度升高显著促进了根际土壤水溶性有机氮(DON)的消耗,导致DOC/DON升高,提高了DON的负根际效应和DOC/DON的正根际效应.干旱胁迫和CO₂浓度升高对土壤总磷脂脂肪酸(总PLFA)和细菌PLFA的根际效应无显著影响.CO₂浓度升高条件下干旱胁迫显著提高了根际土壤G⁺/G^- PLFA,降低了非根际土壤G⁺/G^- PLFA,导致其根际效应显著提高,表明根际微生物群落由自养微生物群落向异养微生物群落的转变.     

短期CO2浓度升高和干旱胁迫对白羊草土壤碳氮和微生物根际效应的影响

采用人工气候室和盆栽控水试验研究黄土丘陵区典型草本植物白羊草在倍增CO₂浓度(800 μmol·mol^-1)下和充分供水(75%～80%的田间持水量)、轻度干旱胁迫(55%～60%的田间持水量)和重度干旱胁迫(35%～40%的田间持水量)下根际和非根际土壤碳氮含量和微生物群落结构及其根际效应.结果表明: CO₂浓度升高和干旱胁迫对白羊草根际和非根际土壤有机碳、全氮和水溶性有机碳(DOC)含量及其根际效应均无显著影响.轻度干旱胁迫下CO₂浓度升高显著促进了根际土壤水溶性有机氮(DON)的消耗,导致DOC/DON升高,提高了DON的负根际效应和DOC/DON的正根际效应.干旱胁迫和CO₂浓度升高对土壤总磷脂脂肪酸(总PLFA)和细菌PLFA的根际效应无显著影响.CO₂浓度升高条件下干旱胁迫显著提高了根际土壤G⁺/G^- PLFA,降低了非根际土壤G⁺/G^- PLFA,导致其根际效应显著提高,表明根际微生物群落由自养微生物群落向异养微生物群落的转变.     

金桂快速繁殖炼苗技术的研究

通过不同生根培养基、炼苗预处理、移栽基质以及叶面施肥等对金桂品种微繁苗炼苗的影响开展试验研究,为目标品种的壮苗快繁提供炼苗的技术支撑。通过对金桂快速繁殖关键技术研究,结果表明,金桂微繁苗生根率和生根数的最佳处理组合为A_3B_2C_3,即B_5基本培养基+2 mg·L^-1NAA+0. 10 mg·L^-16-BA,其生根率、平均生根数分别为:91. 11%、3. 83条/株。炼苗的最佳处理组合为A_3B_2C_3,即以苗圃土∶珍珠岩∶腐殖土=2∶3∶7为炼苗基质,金桂微繁苗炼苗的成活率和苗木增长率分别达到82. 2%、42. 2%。     

Estrogenic activity of triterpene glycosides in yeast two-hybrid assay

Estrogenic potency of six triterpene glycosides, Holothurin A, Holotoxin A₁, Frondoside A, Cucumarioside A₂-2 and Cauloside C, that are natural products and semi-synthesized Ginsenoside-Rh₂, were examined with yeast two-hybrid system, including expressed genes of human estrogen receptor, hER, the co-activator TIF2 and lacZ as a reporter gene. Only Ginsenoside-Rh₂ exhibited significant moderate estrogenic activity in the concentration range of 10⁻⁷ to 10⁻⁶ M. Its effect was approximately 30% of the activity of 17β-estradiol applied at half-effective concentration. This indicates Ginsenosides-Rh₂ is a weak phytoestrogen. The sea cucumber triterpene glycosides, Holothurin A, Holotoxin A₁, Cucumarioside A₂-2 and Frondoside A, and plant glycoside Cauloside C had no appreciable estrogenic activity. Data obtained by yeast two-hybrid assay reflect structure–activity relationship between tested compounds and 17β-estradiol. Only Ginsenoside-Rh₂ has some similarity in chemical structure with 17β-estradiol that might explain affinity of this glycoside to the hER receptor.     

New herbicide-binding site in the photosynthetic electron-transport chain: Competitive herbicide binding at the photosystem I phylloquinone-(vitamin K1)-binding site

The binding of herbicides to the phylloquinone-(primary electron acceptor A₁)-binding site in green plant photosystem (PS) I reaction centers is shown. Dissociation constants (K_d) of various herbicides to the phylloquinone-binding site were estimated by analyzing their competitive inhibition of the reconstitution of the phylloquinone analogue, menadione (vitamin K₃), to the phylloquinone-extracted spinach PS I particles. The phylloquinone-binding site was found to bind o-phenanthroline (K_d = 1.2 × 10⁻⁴ M), but only weak binding was observed with atrazine (K_d > 10⁻² M), although both are known to bind specifically to the quinone-(Q_B)-binding site in reaction centers of purple photosynthetic bacteria or PS II. The inhibitors of the cytochrome b/c₁(ƒ) complex, myxothiazol (K_d=9.5 × 10⁻⁶ M) or antimycin A (K_d = 2.8 × 10⁻⁶ M), also strongly bound to the phylloquinone site. This is the first report showing that the PS I reaction center complex also has a herbicide-binding site, although the site is probably not sensitive in vivo to these herbicides due to its higher affinity for phylloquinone than herbicides. The inhibitor specificity of the PS I phylloquinone site is different from that of the other quinone-functioning sites in the photosynthetic or respiratory electron-transfer chain, suggesting it to have a unique structure.     

Phospholipase A2 in human placental serum

Intervillous blood was collected from term placentae at delivery, and sera were tested for phospholipase A₂ under various experimental conditions. Enzyme activity was found to develop upon extended storage in the cold or at 37°C. The enzyme is reversibly inhibited by dithiothreitol, requires Ca⁺⁺ ions for activity, and tolerates various detergents. The apparent molecular weight is 42 kDa. In all these parameters the serum enzyme behaves similar to the 42 kDa phospholipase A₂ which we recently purified to homogeneity from thoroughly washed placental tissue. Serum phospholipase A₂ appears to be generated by proteolytic processing from a slightly larger inactive precursor which was detected immunochemically. Most likely this protein originates from fetal cells and may be released by membrane damage. We conclude that both placental serum and tissue harbour a novel type of phospholipase A₂ which is distinct from cytosolic and secretory phospholipases A₂. Preference for arachidonate containing substrate suggests a role in eicosanoid production within gestational tissues.     

Characterization of phospholipase A2 and acyltransferase activities in squid (Loligo pealei) axoplasm: comparison with enzyme activities in other neural tissues, axolemma and axoplasmic subfractions.

Phospholipase A₂ and acyltransferase were assayed and characterized in pure axoplasm and neural tissues of squid. Intracellular phospholipase A₂ activity was highest in giant fiber lobe and axoplasm, followed by homogenates from retinal fibers, optic lobe and fin nerve. In most preparations, exogenous calcium (5 mM) caused a slight stimulation of activity. EGTA (2 mM) was somewhat inhibitory, indicating that low levels of endogenous calcium may be required for optimum activity. Phospholipase A₂ was inhibited by 0.1 mM p-bromophenacylbromide, and was completely inactivated following heating.

The level of acylCoA: lysophosphatidylcholine acyltransferase activity was higher in axoplasm and giant fiber lobe than in other neural tissues of the squid. K_m (apparent) and V_max (apparent) for oleoyl-CoA and lysophosphatidylcholine were quite similar for axoplasm and giant fiber lobe enzyme preparations. Acyltransferase activity was inactivated by heat treatment, and greatly inhibited by 0.2 mM p-chloromercuribenzoate, and to a lesser extent by 20 mM N-ethylmaleimide.

Phospholipase A₂ activity was present in fractions enriched in axolemmal membranes (separated from squid retinal fibers and garfish olfactory nerve) from both tissues, and it was also highly concentrated in vesicles derived from squid axoplasm. In all three preparations, phospholipase A₂ activity was stimulated by Ca⁺⁺ (5 mM) and inhibited by EGTA (2 mM). In addition, axoplasmic cytosol (114,000 g supernatant) retained a substantial portion of a Ca⁺⁺-independent phospholipase A₂, active in the presence of 2 mM EGTA. Acyltransferase activity was present at high content in both axolemma membrane rich fractions, and among subaxoplasmic fractions and axoplasmic vesicles.     

Involvement of phospholipase A2 in neurodegeneration

Phospholipases A₂ are a heterogeneous class of enzymes that hydrolyse fatty acids from the sn-2 position of membrane phospholipids. Prolonged stimulation of phospholipase A₂ may damage membrane integrity, not only because of the loss of essential phospholipid from the lipid bilayer but also as a result of an uncontrollable Ca²⁺ influx. The increased levels of intracellular Ca²⁺ may be responsible for enhanced lipolysis, proteolysis and DNA fragmentation. This process along with the accumulation of lipid peroxidation products may be associated with neurodegeneration in acute neural trauma (ischemia, head and spinal cord injuries) and neurodegenerative diseases (Alzheimer's disease).     

Cu(I) and Cu(II) complexes of a pyridine-based pincer ligand

The pincer ligands 2,6-H₃C₅N(CH₂NR₂)₂, L^R, have been studied in their reaction towards CuCl₂ and CuCl. For CuCl₂, the case R=Et gives square-pyramidal (η³-L^Et)CuCl₂ with an apical Cu---Cl distance 0.27 Å longer than the equatorial one. For R=ⁱPr, the chloride-loss product (η³-LⁱPr)CuCl⁺ is established as its CuCl₄ ²⁻ salt. The mer geometry of the ligand in these two compounds is intolerable for Cu(I), and a ligand-redistribution product from CuCl is (η²-L^Me)₂Cu⁺, together with linear CuCl₂ ⁻. Density functional theory (DFT) calculations of monomeric (L^Me)Cu(I)L^q with L=MeCN, C₂H₄ or Cl⁻ show a distinct tendency for one or both NMe₂ arms to dissociate from Cu(I), while the Cu(II) analogs adopt planar geometry.     

Structure-activity relationships of adenosines with heterocyclic N6-substituents

Two series of N⁶-substituted adenosines with monocyclic and bicyclic N⁶ substituents containing a heteroatom were synthesized in good yields. These derivatives were assessed for their affinity ([³H]CPX), potency, and intrinsic activity (cAMP accumulation) at the A₁ adenosine receptor in DDT₁ MF-2 cells. In the monocyclic series, the N⁶-tetrahydrofuran-3-yl and thiolan-3-yl adenosines (1 and 26, respectively) were found to possess similar activities, whereas the corresponding selenium analogue 27 was found to be more potent. A series of nitrogen containing analogues showed varying properties, N⁶-((3R)-1-benzyloxycarbonylpyrrolidin-3-yl)adenosine (30) was the most potent at the A₁AR; IC₅₀ = 3.2 nM. In the bicyclic series, the effect of a 7-azabicyclo[2.2.1]heptan-2-yl substituent in the N⁶-position was explored. N⁶-(7-Azabicyclo[2.2.1]heptan-2-yl)adenosine (38) proved to be a reasonably potent A₁ agonist (K_i = 51 nM, IC₅₀ = 35 nM) while further substitution on the 7″-nitrogen with tert-butoxycarbonyl (31, IC₅₀ = 2.5 nM) and 2-bromobenzyloxycarbonyl (34, IC₅₀ = 9.0 nM) gave highly potent A₁AR agonists.