Chemical studies of viral entry mechanisms: I. Hydrophobic protein-lipid interactions during sendai virus membrane fusion. II. Kinetics of bacteriophage λ DNA injection.

Viruses possess very specific methods of targeting and entering cells. These methods would be extremely useful if they could also be applied to drug delivery, but little is known about the molecular mechanisms of the viral entry process. In order to gain further insight into mechanisms of viral entry, chemical and spectroscopic studies in two systems were conducted, examining hydrophobic protein-lipid interactions during Sendai virus membrane fusion, and the kinetics of bacteriophage λ DNA injection.

Sendai virus glycoprotein interactions with target membranes during the early stages of fusion were examined using time-resolved hydrophobic photoaffinity labeling with the lipid-soluble carbene generator3-(trifluoromethyl)-3-(m-^(125 )I] iodophenyl)diazirine (TID). The probe was incorporated in target membranes prior to virus addition and photolysis. During Sendai virus fusion with liposomes composed of cardiolipin (CL) or phosphatidylserine (PS), the viral fusion (F) protein is preferentially labeled at early time points, supporting the hypothesis that hydrophobic interaction of the fusion peptide at the N-terminus of the F_1 subunit with the target membrane is an initiating event in fusion. Correlation of the hydrophobic interactions with independently monitored fusion kinetics further supports this conclusion. Separation of proteins after labeling shows that the F_1 subunit, containing the putative hydrophobic fusion sequence, is exclusively labeled, and that the F_2 subunit does not participate in fusion. Labeling shows temperature and pH dependence consistent with a need for protein conformational mobility and fusion at neutral pH. Higher amounts of labeling during fusion with CL vesicles than during virus-PS vesicle fusion reflects membrane packing regulation of peptide insertion into target membranes. Labeling of the viral hemagglutinin/neuraminidase (HN) at low pH indicates that HN-mediated fusion is triggered by hydrophobic interactions, after titration of acidic amino acids. HN labeling under nonfusogenic conditions reveals that viral binding may involve hydrophobic as well as electrostatic interactions. Controls for diffusional labeling exclude a major contribution from this source. Labeling during reconstituted Sendai virus envelope-liposome fusion shows that functional reconstitution involves protein retention of the ability to undergo hydrophobic interactions.

Examination of Sendai virus fusion with erythrocyte membranes indicates that hydrophobic interactions also trigger fusion between biological membranes, and that HN binding may involve hydrophobic interactions as well. Labeling of the erythrocyte membranes revealed close membrane association of spectrin, which may play a role in regulating membrane fusion. The data show that hydrophobic fusion protein interaction with both artificial and biological membranes is a triggering event in fusion. Correlation of these results with earlier studies of membrane hydration and fusion kinetics provides a more detailed view of the mechanism of fusion.

The kinetics of DNA injection by bacteriophage λ. into liposomes bearing reconstituted receptors were measured using fluorescence spectroscopy. LamB, the bacteriophage receptor, was extracted from bacteria and reconstituted into liposomes by detergent removal dialysis. The DNA binding fluorophore ethidium bromide was encapsulated in the liposomes during dialysis. Enhanced fluorescence of ethidium bromide upon binding to injected DNA was monitored, and showed that injection is a rapid, one-step process. The bimolecular rate law, determined by the method of initial rates, revealed that injection occurs several times faster than indicated by earlier studies employing indirect assays.

It is hoped that these studies will increase the understanding of the mechanisms of virus entry into cells, and to facilitate the development of virus-mimetic drug delivery strategies.

História de Farrapos: biografia, historiografia e cultura histórica no Rio Grande do Sul oitocentista

Este trabalho procura conhecer os caminhos pelos quais a história do movimento político-militar Farroupilha (1835-1845) foi escrita. Situando essa busca na produção historiográfica do século XIX, optamos por enfatizar o papel da obra biográfica História do General Osório, nesse processo. Escrita por Fernando Luis Osório sobre seu pai, o marquês do Herval, Manoel Luis Osorio e publicada em 1894, o primeiro volume da narrativa biográfica da vida do General Osorio possuí 7 capítulos sobre sua participação nos conturbados quase 10 anos do movimento Farroupilha. É nesses capítulos que concentramos nossa analise, buscando estabelecer o como essa narrativa participou do processo que delegou à Farroupilha um local de destaque na cultura histórica riograndense como experiência histórica valorosa, que lhe permite ser lembrada, narrada e comemorada em festa patriótica até hoje. Dessa forma, procurou-se estabelecer as bases dessa cultura histórica e o papel da Farroupilha junto à ela. Da mesma maneira, visitamos a produção historiográfica do XIX na intenção de compreender seu papel no estabelecimento de valor para o movimento. Analisamos as possibilidades historiografias da obra e as características principais da narrativa como o uso documental, o narrador, as discussões historiográficas e o personagem General Osorio que ali é construído. Identificamos na obra de Fernando Luis Osório impulsos historiográficos característicos do XIX, buscamos relações entre a narrativa ali desenvolvida e a história da Farroupilha e pensamos a participação da mesma no estabelecimento do espaço valoroso dado ao movimento na cultura histórica riograndense de fins do oitocentos.

Estilos naturais: uma bricolagem no Brasil urbano

Esta tese trata da construção de sentidos e significados no estilo natural de alimentação. Parte do princípio que simbolizar é uma forma de organizar a vida, que funde as dimensões do singular e do coletivo, em um corpo socialmente informado. Explora a conceituação e a desnaturalização, nos termos de Luz e de Bourdieu, como estratégias metodológicas, para fundamentar um trabalho de campo com naturistas, veganos e adeptos da alimentação do vivo, todos agrupados na categoria de análise de práticas de alimentação urbana natural. Explora a prática de bricolagem como estratégia de construção simbólica na prática e discute seu efeito ora favorável, ora não, no enfrentamento da crise acirrada pela desigualdade social. Chegamos a três grupos de sentidos na análise interpretativa: o de Natureza re-sacralizada, o de impureza atribuído à carne vermelha e ao de liberdade expresso na reprodução do movimento social urbano denominado contracultura. Todos se apresentam entrelaçados em um sistema de símbolos construídos em um campo específico, segundo regras de um jogo construído também no campo, que confere forças aos agentes, e seguem interesses diversos. Concluímos que uma análise interpretativa contribui na organização das trocas simbólicas, que diferentemente das trocas monetárias, demanda um conhecimento do campo específico e das relações sociais em que são (re)produzidos os sentidos e significados no estilo natural. Consideramos que assim contribuímos para a inserção das ciências sociais no campo da nutrição.

杉木生殖生物学特性的研究

本文就杉木大小孢子、雌雄配子体发育、受精作用、新细胞质形成以及细胞质遗传进行较为详细的研究，主要结果如下： 1.2月底小孢子母细胞进入减数分裂，其核相的变化与通常所描述的一致，细胞器集中排列在赤道板附近，其中质体有一重建的过程。当分裂完成时，在四个核之间产生胼胝质壁，细胞器随机分配在四分体中，每个四分孢子含有线粒体、质体、核糖体、高尔基体、脂体、RER和一些液泡。 2.花粉外壁的形成开始于四分体。颗粒状外壁外层和纤维状的原外壁几乎同时产生，具有三层结构的片层状外壁内层的形成晚于外壁外层。初期外壁外层的物质由小孢子和绒毡层共同提供，后期则主要由绒毡层形成的乌氏体所叠加;外壁内层则由小孢子本身分泌的物质形成，在液泡化之前厚度及数目达到最大，片层之间存在广泛的交叉和融合。3月底花粉成熟，花粉壁由颗粒状外层、片层结构的外壁内层和厚的纤维状内壁所组成，在喙处只有一层外壁内层和内壁。各种细胞器的活动在四分体及游离小孢子时期最为活跃。 3.绒毡层的变化与小孢子的发育密切相关，前者具有发生、发展和解体的过程。期间绒毡层分泌的圆球体在结构上有所不同，减数分裂时为中间电子密度深、外围浅的圆球体结构;四分体及游离小孢子时为中间电子密度浅、外围沉积有孢粉素的乌氏体;成熟花粉时为星芒状的乌氏体，它们共同形成花粉的我壁。绒毡层细胞中的细胞器也经历一个变化过程，造粉体的含量在四分体时达到高潮，在小孢子液泡期时基本消耗殆尽并解体;大量的粗糙内质网成零分布在质膜处，并与质膜走向平行。绒毡层解体时出现特殊的现象，具多个由多层膜组成的吞噬泡，并成堆分布，RER和线粒体为最后解体的细胞器。绒毡层外切向面的周绒毡层膜由两层膜组成，其上分布有大量的孢粉素和乌氏体。 4.萌发的花粉管4月底穿过珠心顶端，内含两个不育核和一精原细胞，不育核分布在具有淀粉、脂滴、线粒体和RER的原花粉细胞质中，位于精原细胞之前，这种关系保持到受精前。6月初精原细胞体积迅速增加到最大，相应的细胞质也发生变化，以线粒体为主的细胞质中出现少量质体，线粒体再资分化，细胞质中含有大量的可溶性多糖。 5.受精前，精原细胞分裂产生两个大小和形状相同的精细胞，彼此之间由胞间连丝的横壁联系在一起，并被原花粉细胞质所包被。精细胞中质体和淀粉粒的数量大幅度增加。精核们于细胞的中央，细胞质呈现区域分布的特点，据所含细胞器的差别，人核膜向外，细胞质分为五层，中间两层最为突出，即线粒体层和淀粉层，其间有大量核糖体的存在，线粒体层位于淀粉层的里面。这两层占据精细胞的大部分体积。 6.雌配子体游离核时期持续时间较长，近两个月，而从细胞化到卵细胞成熟则非常迅速，大约只需两周左右的时间。进一步发育，颈卵器中的液泡减少，细胞质变浓。初期少量的质体和淀粉粒被膨大的内质网片段和小泡的融合而与细胞质相隔，并最终退化。小内含物增加，亲锇颗粒出现。 7.成熟的卵细胞中具有大量各种形式的小内含物，细胞质被平行和环形的内质网所分隔。核膜外围有一圈疏松排列的亲锇颗粒，核仁变为多个基本为圆形的小核仁。没有质体的存在，大量脱分化的线粒体和核糖体集中分布在卵核的下部。 8.受精作用主要发生在6.9-6.16日期间，雄性细胞质始终伴随着精核向卵核移动，当两核接近时，朝向精核－面的卵核形成凹陷内，性细胞质覆盖在精核之上并最终包围两性核，而把雌性细胞质排除在外。因此，受精卵周围的胞质主要为由质体、线粒体和核糖体组成的新细胞质，且线粒体和质体的分布形式与精细胞的相同，即线粒体在里层，质体在外层。 9.融合后的合子核随即进行有丝分裂，形成两个原胚游离核，两个游离核同步分裂，并向基部移动，游离核始终分布在新细胞质中。八游离核时形成细胞壁，原胚属标准型。胚细胞中基本不含淀粉粒，具有大量的线粒体、高尔基体、RER和原质体。原胚之上的卵细胞质退化解体。杉木的质体和线粒体均为父本遗传。 10.幼胚的胚性细胞和胚柄细胞具有明显的差别，主要表现在质体的存在形式和高尔基体的数量上。胚性细胞中，原质体分布在核周围，大量的RER、高尔基体和线粒体平行于胚的走向;位于其上的胚柄细胞则含有淀粉粒和特别多的高尔基体。具简单多胚和裂生多胚，胚柄系统发达，7月底出现根原始细胞，8月中旬胚分化完成，具有两个子叶。

水松生殖生物学的研究

本文运用组织化学技术、透射电镜技术，就水松雌雄配子体发育，精卵细胞形成和结构，受精作用以及细胞质遗传等问题进行了较为详细的研究。主要结果如下： 1 绒毡层发育与雄配子体发育，尤其与花粉外壁的形成关系密切。绒毡层组织在小孢子母细胞和减数分裂时期达到发育高峰。四分体阶段和游离小孢子发育前期，绒毡层细胞中内质网、脂体、线粒体非常活跃，参与孢粉素和乌氏体的形成和转移。乌氏体有球形、星芒状两种类型，它们主要参与花粉外壁内层和外壁外层的建成。小孢子发育后期绒毡层细胞开始明显解体。 2 减数分裂阶段，质体、线粒俺等细胞器和造粉体随细胞核和染色体的变化，出现有规律的迁移现象。减数分裂前期，细胞核移到细胞的一侧;各种细胞器和造粉体迁移到细胞的另一侧，并随染色体移向细胞中部而转移到细胞质周缘。减数分裂中期l和后期，各种细胞器和造粉体汇聚在赤道板丙侧分布。说明细胞质中微管系统在起调节作用。 3 花粉壁形成开始于四分体时期。片层状结构的外壁内层随纤维状原外壁的出现雨形成。外壁外层和花粉内壁在小孢子发育中期几乎同时形成。小孢子细胞和绒毡层组织共同参与了外壁内层和外壁外层孢粉素物质的合成、转运。水松成熟花粉由孢粉素组成的外壁外层、片层状结构的外壁内层及有分层状结构的内壁构成。 4 传粉到受精间隔要四个月左右的时间。六月上旬，花粉管和精原细胞抵达颈卵器上部。精原细胞富含淀粉粒、质体、线粒体和异形泡并分区分布。受精前，精原细胞分裂形成两个大小和形状相同豹精细胞。精细胞含有质体、线粒体和异型泡等细胞器。 5 雌配子体游离核持续时间长，从颈卵器原始细胞形成到卵细胞发育成熟所需时间较短。复合颈卵器结构，颈细胞和套层细胞形态结构特殊。中央细胞不经分裂，直接行使卵细胞的功能。 6 成熟卵细胞中除弋量各种形式的内含物外，细胞质被庞大的内质网包围、分割，形成网膜系统。质体、淀粉等细胞器被包含成大内含物。在卵核周围细胞质中没有发现质体和发育完好的线粒体存在。 7 受精作用发生在六月十日左右。雌雄核融合时，朝向精核一面的卵核形成凹陷，精核陷入其中与卵核进一步融合。精卵融合可发生在颈卵器的上部、中部、下部甚至底部。有旋转受精、两个精核同时与一个卵核受精及多卵细胞现象等。 8 新细胞质主要为雄性细胞质成分。合子转移到颈卵器基部分裂，形成原胚游离核。合子、新细胞质和游离核周围淀粉鞘显著。八游离核时期形成细胞壁，新细胞质和淀粉粒转移到原胚细胞中。 9 水松的质体和线粒体为父系遗传。

Vortex filament simulation of trailing vortex merger

Aircraft in high-lift configuration shed multiple vortices. These generally merge to form a downstream wake consisting of two counter-rotating vortices of equal strength. The understanding of the merger of two co-rotating trailing vortices is important in evaluating the separation criteria for different aircraft to prevent wake vortex hazards during landing and take-off. There is no existing theoretical method on the basis of which such norms can be set. The present study is aimed at gaining a better understanding of the behaviour of wake vortices behind the aircraft. Two dimensional studies are carried out using the vortex blob method and compared with Bertenyi's experiment. It is shown that inviscid two dimensional effects are insufficient to explain the observations. Three dimensional studies, using the vortex filament method, are applied to the same test case. Two Lamb-Oseen profile vortices of the same dimensions and initial separation as the experiment are allowed to evolve from a straight starting condition until a converged steady flow is achieved. The results obtained show good agreement with the experimental distance to merger. Core radius and separation behaviour is qualitatively similar to experiment, with the exception of rapid increases at first. This may be partially attributable to the choice of filament-element length, and recommended further work includes a convergence study for this parameter. Copyright © 2005 by the American Institute of Aeronautics and Astronautics, Inc. All rights reserved.

Spontaneous emission from radiative chiral nematic liquid crystals at the photonic band-gap edge: An investigation into the role of the density of photon states near resonance

In this article, we investigate the spontaneous emission properties of radiating molecules embedded in a chiral nematic liquid crystal, under the assumption that the electronic transition frequency is close to the photonic edge mode of the structure, i.e., at resonance. We take into account the transition broadening and the decay of electromagnetic field modes supported by the so-called "mirrorless"cavity. We employ the Jaynes-Cummings Hamiltonian to describe the electron interaction with the electromagnetic field, focusing on the mode with the diffracting polarization in the chiral nematic layer. As known in these structures, the density of photon states, calculated via the Wigner method, has distinct peaks on either side of the photonic band gap, which manifests itself as a considerable modification of the emission spectrum. We demonstrate that, near resonance, there are notable differences between the behavior of the density of states and the spontaneous emission profile of these structures. In addition, we examine in some detail the case of the logarithmic peak exhibited in the density of states in two-dimensional photonic structures and obtain analytic relations for the Lamb shift and the broadening of the atomic transition in the emission spectrum. The dynamical behavior of the atom-field system is described by a system of two first-order differential equations, solved using the Green's-function method and the Fourier transform. The emission spectra are then calculated and compared with experimental data. © 2013 American Physical Society.

Spontaneous emission from radiative chiral nematic liquid crystals at the photonic band-gap edge: an investigation into the role of the density of photon states near resonance.

In this article, we investigate the spontaneous emission properties of radiating molecules embedded in a chiral nematic liquid crystal, under the assumption that the electronic transition frequency is close to the photonic edge mode of the structure, i.e., at resonance. We take into account the transition broadening and the decay of electromagnetic field modes supported by the so-called "mirrorless"cavity. We employ the Jaynes-Cummings Hamiltonian to describe the electron interaction with the electromagnetic field, focusing on the mode with the diffracting polarization in the chiral nematic layer. As known in these structures, the density of photon states, calculated via the Wigner method, has distinct peaks on either side of the photonic band gap, which manifests itself as a considerable modification of the emission spectrum. We demonstrate that, near resonance, there are notable differences between the behavior of the density of states and the spontaneous emission profile of these structures. In addition, we examine in some detail the case of the logarithmic peak exhibited in the density of states in two-dimensional photonic structures and obtain analytic relations for the Lamb shift and the broadening of the atomic transition in the emission spectrum. The dynamical behavior of the atom-field system is described by a system of two first-order differential equations, solved using the Green's-function method and the Fourier transform. The emission spectra are then calculated and compared with experimental data.

日本落叶松微体繁殖及植株再生的研究

本文以10、13、29年生的日本落叶松(Laxix kaempferi (Lamb.)Carr.) 朝85；38、朝6号无性系的冬芽为材料，经过腑芽诱导、不定茎伸长、生根诱导和培养等阶段，形成了再生植株。文中主要研究了基本培养基种类、激素组合及其浓度、遗传材料（无性系）、外植体年龄、活性炭对芽诱导、茎伸长以及继代、温度、染菌等对植株再生过程的影响。通过实验，选择出微体繁殖过程中茎诱导、茎伸长、生根诱导等各个阶段的优化培养基，它们分别是：茎诱导SH+0.5mg/l Zea+0.05mg/l IAA WPM+1mg/l zea+1mg/l IAA；茎伸长 改良MS+0.1mg/l IBA+0.5mg/l NAA+1% AC; 根诱导 改良MS+0.2mg/l IBA+0.1mg/l NAA+0.15% AC。培养条件诱导、伸长时光照1000－3000Lux，温度 25±1℃；生根时光照不变，温度 20±1℃。对影响外植体发育和器官发生各种因素的研究结果表明：继代可明显提高诱导率，继代、低温处理相结合促进生根；高浓度（1%）活性炭对茎伸长有明显的促进作用，低浓度（0.15）活性炭则促进根的形成；诱导率也随年龄、无性系的变化而不同；在根诱导阶段，细感染并不影响生根。同时，文中还对无菌幼苗的扦插进行了实验，以为成龄树木棰根提供可借鉴的资料。

次生植被土壤碳、氮养分季节动态及凋落物输入与温度的影响

土壤微生物量、可溶性有机碳与氮虽然只占土壤有机碳、氮总量的较小部分，但可以在土壤全碳、氮变化之前反映土壤微小的变化，又直接参与土壤生物化学转化过程，因而在植被恢复过程中，较其它土壤理化性质等能够更好地指示土壤恢复情况。在青藏高原东缘存在大面积的次生人工林替代灌丛或采伐迹地，而关于这些人工林替代后的生态效果和生态过程的评估却十分缺乏，本研究通过评估岷江上游植被恢复重建过程中典型人工替代次生植被凋落物层与土壤碳、氮等养分大小，动态监测土壤微生物生物量、水溶性碳、氮等指标，结合温度与凋落物输入等影响土壤活性有机碳、氮因子的控制试验，系统分析不同人工替代次生植被土壤碳、氮等养分的差异原因，试图寻找低效人工林优化调控与持续管理技术，为区域生态公益林持续管理提供理论和技术依据。主要结论如下： 1. 通过对不同人工替代次生植被凋落物层和土壤碳、氮分析发现，油松和华山松人工林替代次生灌丛后土壤碳、氮含量较灌丛和阔叶人工林低，主要原因可能为凋落物质量(C/N)较差，而引起碳、氮等元素难以归还土壤。进而通过对不同人工替代次生植被凋落物层和土壤微生物生物量、水溶性有机碳、氮等指标的季节性动态模式的分析，发现各次生植被土壤微生物生物量C、N，P以及土壤水溶性碳、氮含量均呈明显季节性动态，呈现秋季明显大于其它季节，冬季最低，在表层土壤最为明显。 2. 油松、华山松人工林凋落物层和土壤水溶性有机碳(WDOC)、土壤水溶性有机氮(WDON)明显低于灌丛和连香树，土壤微生物生物量C、N也以油松和华山松人工林最低，而落叶类植被，如灌丛、连香树和落叶松之间没有明显差异，说明可利用底物的数量和质量差异是影响各次生植被凋落物分解和土壤微生物活性的主要原因。MBC/OC和MBN/ON能较好地指示土壤微生物活性的变化，MBC/OC凋落层总体以灌丛和连香树人工林最高，油松和华山松人工林最低；而土壤中MBC/OC连香树人工最高，华山松人工林最低。说明以油松和华山松为主的人工造林替代乡土阔叶灌丛造成土壤C、N等养分严重匮乏，微生物活性低下是影响其养分周转的主要原因。 3. 从各次生植被凋落物产生看，凋落物年归还量最大的为华山松人工林(5.1×103 kg ha-1)，其次为落叶松人工林(4.8×103 kg ha-1)，阔叶灌丛林地凋落物产生总量(4.4×103 kg ha-1)略大于油松人工林(4.2×103 kg ha-1)，最小的为连香树人工林(3.6×103 kg ha-1)；叶是凋落物的主体，落叶类树种月动态表现为单峰型，高峰主要在10-11月，如落叶松、连香树和灌丛林；常绿的松类月动态不明显，各月基本相同，最为明显地为油松林，华山松人工林略有二个小峰，分别出现在11月和5月。落叶阔叶灌丛的凋落物分解速率大于常绿针叶林，如油松和华山松。结合凋落物的产生量和分解速率，不同树种人工林替代次生阔叶灌丛后，人工油松和华山松林枯落物总贮量和厚度明显大于落叶松人工林、灌丛林和连香树人工林，说明以油松和华山松为主的人工造林替代乡土阔叶灌丛延缓了有机物向土壤的顺利归还，不利于土壤C、N等养分循环。 4. 通过控制地面凋落物和地下根系输入有机物对土壤碳、氮的影响研究发现，(1) 单独去除根系以及根系与地面凋落物同时去除处理1年后对表层(0-10cm)土壤WDOC均没有显著影响，而土壤WDON显著增加，油松人工林土壤微生物生物量C、N显著降低，人工落叶松林没有显著差异，说明油松人工林土壤微生物活性对地下碳输入的依赖大于其它次生植被，而落叶松土壤微生物活性对地下碳输入依赖性较小；去除地面凋落物，明显降低了落叶松人工林土壤WDOC，华山松和连香树土壤WDON均较对照显著减少，油松人工林土壤微生物量C较对照显著减少；双倍增加地面凋落物处理对土壤微生物生物量、WDOC和WDON没有明显地增加，相反，连香树、华山松和油松人工林土壤WDON较对照减少。说明油松人工林微生物活性不仅依赖于地下碳输入，而且对地上有机物输入的依赖性也较大；连香树、落叶松和华山松人工林土壤微生物生物量并没有因地面凋落物的去除减少可能与土壤总有机碳含量及活性均较高有关，而双倍增加地面凋落物反而降低了土壤微生物生物量，说明凋落物覆盖后改变了土壤微气候。 5. 碳矿化累积量与有机碳含量和活性有机碳含量之间存在显著地正相关关系。凋落物碳累积矿化量、矿化速率以连香树最高，油松和华山松人工林次之，落叶阔叶灌丛低于常绿针叶纯林，导致其差异的主要原因可能为凋落物产生的时间动态模式不一样，致使凋落物起始分解时间不一致。而土壤层有机碳矿化速率和矿化量以阔叶落叶灌丛和连香树最高，油松和华山松人工土壤最低，再次证实利用针叶纯林恢复植被阻碍了有机质周转与循环。 6. 凋落物累积矿化量与C/N值呈显著地相关关系，并随着温度的升高而明显增加，而土壤累积矿化量与C/N值没有显著相关关系，说明土壤有机碳质量(C/N)对温度的响应不十分明显。通过双指数模型对不同温度下碳矿化过程进行模拟和计算出活性有机碳与惰性有机碳比例，发现温度升高促进了惰性有机碳向活性有机碳的转化，增加了活性有机碳含量，说明温度升高可促进次生植被凋落物与土壤有机质的分解，进而可影响到林地碳源/汇关系的变化。 综上，通过对不同人工替代次生植被凋落物与土壤C、N大小、以及土壤微生物生物量、水溶性C、N等指标动态变化模式研究，结合温度与凋落物数量输入等影响土壤活性C、N因子的综合分析，以油松和华山松人工纯林对山地植被恢复，延缓或阻碍了有机质周转与循环，造成了土壤肥力退化。对现有低效人工纯林改造，应为地面大量有机物分解创造条件。 Although soil microbial biomass, dissolved organic carbon (DOC) and dissolved organic nitrogen (DON) are a small part of total soil organic carbon and nitrogen, they can directly participate in the process of soil biochemical translation and indicate the fine changes before changes of soil total organic carbon and nitrogen occur. So, they are good indexes to indicate soil restoration condition during the process of vegetation rehabilitation. There are large areas of secondary vegetations which substitute for indigenous shrubs in the eastern fringe of Qinghai-Tibet Plateau. However, it is not well known that the ecological effect and process after substitution by different secondary plantations. Based on comparison of soil organic and nitrogen contents in litter layer and soil under different secondary vegetations in upper reaches of Minjiang River, soil microbial biomass, DOC and DON in litter layer and soil were investigated in order to analyze the seasonal dynamic. Combining the effects of temperature, litter input and root exclusion on soil microbial biomass, DOC and DON, we also aim to understand the reason and mechanism of difference in soil carbon and nitrogen contents among different secondary vegetations. The study would contribute to comprehensively understanding C and N cycling processes and provide optimal control and sustainable technology of low-effect plantations in these regions. The results are as follows: (1) Organic carbon and nitrogen in litter layers and soil under different substitution plantations were investigated. The results showed that contents of soil organic carbon and nitrogen were lower in P. tabulaeformis (PT) and P. armandi Franch(PA) than those in native broad-leaf shrub and broad-leaf plantation. The low quality (C/N) of litter in PT and PA plantations caused carbon and nitrogen returning to soil difficultly. Seasonal dynamic of soil microbial carbon (MBC),-nitrogen (MBN),-phosphor (MBP), and WDOC and WDON showed similar pattern, which had the highest values in autumn and the lowest values in winter. (2) WDOC and WDON in litter layers and soil under PT and PA plantations were significantly lower than those in native broad-leaf shrub and Cercidiphyllum japonicum Sieb. et Zucc.(CJ). Soil MBC and MBN were also the lowest, while there were no significant differences among deciduous vegetations, i.e. native broad-leaf shrub, CJ and Larix kaempferi Lamb.(LK) plantation. The results suggested that difference in quantity and quality of available substance was main reason that affected the activity of microbe in soil and litter layer. MBC/OC and MBN/ON were good indexes to indicate the change of soil microbial activity. MBC/OC of litter had the highest value under native broad-leaf shrub and CJ plantation, and had the lowest value in PT and PA plantations, while MBC/OC of soil was the highest under CJ plantation, and was the lowest in PT and PA plantations. These results indicated that PT and PA plantations substituting for native broad-leaf shrub caused deficit of carbon and nitrogen in soil, low microbial activity was a main reason influencing the cycling and turnover of carbon and nitrogen in soil. (3) The annual litter fall production, composition, seasonal dynamic and decomposition of five typical secondary stands in upper reaches of Minjiang River were studied in this paper. The annual litter productions were: PA (5.1×103 kg ha-1), LK(4.8×103 kg ha-1), native broad-leaf shrub (4.4×103 kg ha-1), PT(4.2×103 kg ha-1)，CJ(3.6×103 kg ha-1). The litter production of leaves in five secondary vegetations occupied a higher percentage in the annual total litter production than those of other components. The litterfall was mostly producted in the cool and dry period (October-November) for deciduous vegetations and relatively equably producted in every season for evergreen coniferous vegetations. The decomposition rate of leaf litter in the broad-leaf stand was higher than those in evergreen coniferous stand. Combined with annual litter fall production and decomposition rate of leaf litter, we found that stock and depth of litter layer were significantly larger in PT and PA plantations than those in native broad-leaf shrub, LK and CJ plantations. The results confirmed that PT and PA plantations substituting for native broad-leaf shrub delayed organic matter returning to soil and hindered cycling of carbon and nitrogen again. (4) We explored plant litter removal, double litter addition, root trenching, and combining root trenching and litter removal treatments to examine the effects of above- and belowground carbon inputs on soil microbial biomass, WDOC and WDON in four secondary plantations. During the experimental period from June 2007 to July 2008, 1 year after initiation of the treatments, WDOC in soil did not vary in root trenching, and combining root trenching and litter removal treatments, while WDON in soil significantly increased compared with CK treatment. Root trenching reduced soil MBC and MBN in PT plantation, while MBC and MBN in soil did not vary in LK plantation. The rasults implied that soil microbial activity was more dependent on belowground carbon input in PT plantation than those in other secondary plantations, on the contrary, soil microbial activity in LK plantation was not dependent on belowground carbon input. Plant litter removal significantly decreased soil WDOC in LK plantation, decreased WDON in PA and CJ plantations, and also significantly reduced soil MBC in PT plantation. However, double litter addition did not increase soil microbial biomass, WDOC and WDON, on the contrary, soil WDON in CJ, PA and PT plantations were decreased. These suggested that soil microbial activity was not only dependent on belowground carbon input, but also on aboveground organic material input. Double litter addition could change the microclimate and result in the decrease of soil microbial activity in CJ, PA and PT plantations. (5) We measured carbon mineralization in a 107 days incubation experiment in 5℃,15℃ and 25℃. Carbon cumulative mineralization was positively correlated with organic matter and labile organic carbon in litter layer and soil. Cumulative carbon mineralization and mineralization rate of litter layers in PT and PA plantations were higher than that in native broad-leaf shrub. This difference between native broad-leaf shrub and coniferous plantations in cumulative carbon mineralization and mineralization rate of litter layers could be attributed to the initiating time of decomposition due to the difference in seasonal dynamic of litter fall production between two types of secondary plantations. However, cumulative carbon mineralization and mineralization rate in soil were the highest in native broad-leaf shrub and CJ plantation, and were the lowest in PT and PA plantations. This also confirmed that PT and PA plantations substituting for native broad-leaf shrub hindered the cycling and turnover of organic matter again. (6) Carbon cumulative mineralization was positively correlated with C/N in litter layer and increased with temperature increasing, while carbon cumulative mineralization was not correlated with C/N in soil. This indicated that soil organic matter quality (C/N) was insensitive to temperature. Applying bi-exponential model, we computed the percent of labile and stable carbon in different temperature incubation and found that temperature increasing would accelerate the transform from stable carbon to labile carbon and increase the percentage of labile organic carbon. This illuminated that temperature incraesing could facilitate the decomposition of litter and soil organic matter in secondary vegetations and hence affect the relationship between carbon source and sink.

Studied on the interactions of antibody-porphyrin by circular dichroism and ultraviolet-visible spectrophotometry

Circular dichoism and UV-vis measurements were used to study the interaction between porphyrin and monoclonal antibodies ( McAbs). McAbs-porphyrin complex formation is usually accompanied by significant bathochromic shift and hyperchromicity changes of the absorption maxima in the porphyrin soret band region. Induced CD spectra in the same region (350 similar to 450 nm) were detected upon complex formation. They follow Lamb-Beer's law and exhibit isosbestic behavior. Both the UV-Vis and induced CD spectra of the antibody: porphyrin complex remain unchanged over a broad pH range ( pH 6 similar to 11), indicating remarkable stability of the complex and reflecting the dominant role of hydrophobic interaction between the hapten benzophenone and the antibody combining site.