Microstructure control and iodine doping of bismuth telluride

On the materials scale, thermoelectric efficiency is defined by the dimensionless figure of merit zT. This value is made up of three material components in the form zT = Tα²/ρκ, where α is the Seebeck coefficient, ρ is the electrical resistivity, and κ is the total thermal conductivity. Therefore, in order to improve zT would require the reduction of κ and ρ while increasing α. However due to the inter-relation of the electrical and thermal properties of materials, typical routes to thermoelectric enhancement come in one of two forms. The first is to isolate the electronic properties and increase α without negatively affecting ρ. Techniques like electron filtering, quantum confinement, and density of states distortions have been proposed to enhance the Seebeck coefficient in thermoelectric materials. However, it has been difficult to prove the efficacy of these techniques. More recently efforts to manipulate the band degeneracy in semiconductors has been explored as a means to enhance α.

The other route to thermoelectric enhancement is through minimizing the thermal conductivity, κ. More specifically, thermal conductivity can be broken into two parts, an electronic and lattice term, κ_e and κ_l respectively. From a functional materials standpoint, the reduction in lattice thermal conductivity should have a minimal effect on the electronic properties. Most routes incorporate techniques that focus on the reduction of the lattice thermal conductivity. The components that make up κ_l (κ_l = 1/3Cνl) are the heat capacity (C), phonon group velocity (ν), and phonon mean free path (l). Since the difficulty is extreme in altering the heat capacity and group velocity, the phonon mean free path is most often the source of reduction.

Past routes to decreasing the phonon mean free path has been by alloying and grain size reduction. However, in these techniques the electron mobility is often negatively affected because in alloying any perturbation to the periodic potential can cause additional adverse carrier scattering. Grain size reduction has been another successful route to enhancing zT because of the significant difference in electron and phonon mean free paths. However, grain size reduction is erratic in anisotropic materials due to the orientation dependent transport properties. However, microstructure formation in both equilibrium and nonequilibrium processing routines can be used to effectively reduce the phonon mean free path as a route to enhance the figure of merit.

This work starts with a discussion of several different deliberate microstructure varieties. Control of the morphology and finally structure size and spacing is discussed at length. Since the material example used throughout this thesis is anisotropic a short primer on zone melting is presented as an effective route to growing homogeneous and oriented polycrystalline material. The resulting microstructure formation and control is presented specifically in the case of In₂Te₃-Bi₂Te₃ composites and the transport properties pertinent to thermoelectric materials is presented. Finally, the transport and discussion of iodine doped Bi₂Te₃ is presented as a re-evaluation of the literature data and what is known today.

Design of an aerial tramway

An Aerial Tramway is a device for transporting material in receptacles over wire ropes supported at various elevations above the ground by means of posts or standards. Wire rope has been known for many centuries, and there is a drawing of a ropeway appearing in a book dated 1411.

Geology of the Pacoima Hills

The Pacoima Hills lie between Foothill Boulevard and the San Fernando Road, three miles southeast of San Fernando, California. In this area are exposed Jurassic(?) granodiorite intruded in older gneiss, and a mid Miocene Topango (?) sedimentary section lying in both fault and sedimentary contact with the intrusive complex. Two distinct lava flows and a small laccolith of andesite occur within the Topango (?) formation. The principal structural feature is an anticline plunging steeply northward. An upward acting force is postulated to have produced this anticline; upon cessation of the force, normal faulting occurred with consequent down-dropping of north-south blocks.

On an embedding property of generalized Carter subgroups

If E and F are saturated formations, we say that E is strongly contained in F if for any solvable group G with E-subgroup, E, and F-subgroup, F, some conjugate of E is contained in F. In this paper, we investigate the problem of finding the formations which strongly contain a fixed saturated formation E.

Our main results are restricted to formations, E, such that E = {G|G/F(G) ϵT}, where T is a non-empty formation of solvable groups, and F(G) is the Fitting subgroup of G. If T consists only of the identity, then E=N, the class of nilpotent groups, and for any solvable group, G, the N-subgroups of G are the Carter subgroups of G.

We give a characterization of strong containment which depends only on the formations E, and F. From this characterization, we prove:

If T is a non-empty formation of solvable groups, E = {G|G/F(G) ϵT}, and E is strongly contained in F, then

(1) there is a formation V such that F = {G|G/F(G) ϵV}.

(2) If for each prime p, we assume that T does not contain the class, S_p’, of all solvable p’-groups, then either E = F, or F contains all solvable groups.

This solves the problem for the Carter subgroups.

We prove the following result to show that the hypothesis of (2) is not redundant:

If R = {G|G/F(G) ϵS_r’}, then there are infinitely many formations which strongly contain R.

The interactions of basic proteins and DNA

I. ELECTROPHORESIS OF THE NUCLEIC ACIDS

A zone electrophoresis apparatus using ultraviolet optics has been constructed to study nucleic acids at concentrations less than 0.004%. Native DNA has a mobility about 15% higher than denatured DNA over a range of conditions. Otherwise, the electrophoretic mobility is independent of molecular weight, base composition or source. DNA mobilities change in the expected way with pH but the fractional change in mobility is less than the calculated change in charge. A small decrease in mobility accompanies an increase in ionic strength. RNA’s from various sources have mobilities slightly lower than denatured DNA except for s-RNA which travels slightly faster. The important considerations governing the mobility of nucleic acids appear to be the nature of the hydrodynamic segment, and the binding of counterions. The differences between electrophoresis and sedimentation stem from the fact that all random coil polyelectrolytes are fundamentally free draining in electrophoresis.

II. THE CYTOCHROME C/DNA COMPLEX

The basic protein, cytochrome c, has been complexed to DNA. Up to a cytochrome:DNA mass ratio of 2, a single type of complex is formed. Dissociation of this complex occurs between 0.05F and 0.1F NaCl. The complexing of cytochrome to DNA causes a slight increase in the melting temperature of the DNA, and a reduction of the electrophoretic mobility proportional to the decrease in net charge. Above a cytochrome:DNA mass ratio of 2.5, a different type of complex is formed. The results suggest that complexes such as are formed in the Kleinschmidt technique of electron microscopy would not exist in bulk solution and are exclusively film phenomena.

III. STUDIES OF THE ELECTROPHORESIS AND MELTING BEHAVIOUR OF NUCLEOHISTONES

Electrophoresis studies on reconstituted nucleohistones indicate that the electrophoretic mobility for these complexes is a function of the net charge of the complex. The mobility is therefore dependent on the charge density of the histone complexing the DNA, as well as on the histone/DNA ratio. It is found that the different histones affect the transition from native to denatured DNA in different ways. It appears that histone I is exchanging quite rapidly between DNA molecules in 0.01 F salt, while histone II is irreversibly bound. Histone III-IV enhances the capacity of non-strand separated denatured DNA to reanneal. Studies on native nucleoproteins indicate that there are no gene-sized uncomplexed DNA regions in any preparations studied.

IV. THE DISSOCIATION OF HISTONE FROM CALF THYMUS CROMATIN

Calf thymus nucleoprotein was treated with varying concentrations of NaCl. The identity of the histones associated and dissociated from the DNA at each salt concentration was determined by gel electrophoresis. It was found that there is no appreciable histone dissociation below 0.4 F NaCl. The lysine rich histones dissociate between 0.4 and 0.5 F NaCl. Their dissociation is accompanies by a marked increase in the solubility of the chromatin. The moderately lysine rich histones dissociate mainly between 0.8 and 1.1 F NaCl. There are two arginine rich histone components: the first dissociates between 0.8 F and 1.1 F NaCl, but the second class is the very last to be dissociated from the DNA (dissociation beginning at 1.0 F NaCl). By 2.0 F NaCl, essentially all the histones are dissociated.

The properties of the extracted nucleoprotein were studied. The electrophoretic mobility increases and the melting temperature decreases as more histones are dissociated from the DNA. A comparison with the dissociation of histones from DNA in NaClO₄ shows that to dissociate the same class of histones, the concentration of NaCl required is twice that of NaClO₄.

Development of a sampling expert system:"FISHMAP"

This research program consisted of three major component areas: (I) development of experimental design, (II) calibration of the trawl design, and (III) development of the foundation for stock assessment analysis. The products which have I. EXPERIMENTAL DESIGN resulted from - the program are indicated below: The study was successful in identifying spatial and temporal distribution characteristics of the several key species, and the relationships between given species catches and environmental and physical factors which are thought to influence species abundance by areas within the mainstem of the Chesapeake Bay and tributaries

Studies of dinucleoside monophosphates and monomer-polynucleotide interactions by proton magnetic resonance

The nature of the intra- and intermolecular base-stacking interactions involving several dinucleoside monophosphates in aqueous solution have been investigated by proton magnetic resonance spectrosocopy, and this method has been applied to a study of the interaction of polyuridylic acid with purine and adenosine monomers.

The pmr spectra of adenylyl (3' → 5') cytidine (ApC) and cytidylyl (3' → 5') adenosine (CpA) have been studied as a function of concentration and temperature. The results of these studies indicate that the intramolecular base-stacking interactions between the adenine and cytosine bases of these dinucleoside monophosphates are rather strong, and that the stacking tendencies are comparable for the two sequence isomers. The chemical shifts of the cytosine H₅ and adenine H₂ protons, and their variations with temperature, were shown to be consistent with stacked conformations in which both bases of the dinucleoside monophosphates are preferentially oriented in the anti conformation as in similar dApdC, and dCpdA (dA = deoxyadenosine; dC = deoxycytidine) segments in double helical DNA. The intramolecular stacking interaction was found to have a pronounced effect on the conformations of the ribose moieties, and these conformational changes are discussed. The concentration studies indicate extensive self-association of these dinucleoside monophosphates, and analysis of the concentration data facilitated determination of the dimerization constant for the association process as well as the nature of the intermolecular complexes.

The dependence of the ribose conformation upon the extent of intramolecular base-stacking was used to demonstrate that the base-base interaction in cytidylyl (3' → 5') cytidine (CpC) is rather strong, while there appears to be little interaction between the two uracil bases of uridylyl (3' → 5') uridine (UpU).

Studies of the binding of purine to several ribose and deoxyribose dinucleoside monophosphates show that the mode of interaction is base-stacking, and evidence for the formation of a purine-dinucleoside monophosphate intercalated complex is presented. The purine proton resonances are markedly broadened in this complex, and estimates of the purine linewidths in the complex and the equilibrium constant for purine intercalation are obtained.

A study of the interaction of unsubstitued purine with polyuridylic acid at 29°C by pmr indicated that purine binds to the uracil bases of the polymer by base-stacking. The severe broadening of the purine proton resonances observed provides strong evidence for the intercalation of purine between adjacent uracil bases of poly U. This interaction does not result in a more rigid or ordered structure for the polymer.

Investigation of the interaction between adenosine and polyuridylic acid revealed two modes of interaction between the monomer and the polymer, depending on the temperature. At temperatures above 26°C or so, monomeric adenosine binds to poly U by noncooperative A-U base stacking. Below this temperature, a rigid triple-stranded 1A:2U complex is formed, presumably via cooperative hydrogen-bonding as has previously been reported.

These results clearly illustrate the importance of base-stacking in non-specific interactions between bases, nucleosides and nucleotides, and also reveal the important role of the base-stacking interactions in cooperatively for med structures involving specific base-pairing where both types of interaction are possible.

Evaluation of NRA tracking studies. Future projects and technical development

Fish tracking is a valuable technique for the provision of detailed information on the behaviour patterns of individual fish especially during estuarine and riverine migration. 2. Tracking studies help in the provision of a comprehensive description of the variety offish behaviour patterns in response to factors such as water flow, obstructions and water quality. 3. There are advantages to be gained by complementing fish tracking studies with data collected from fish counters and vice versa. 4. An overall evaluation of NRA fish tracking projects is presented in the wider context of NRA strategic research objectives. 5. The requirement for future development of tracking equipment, improved data analysis techniques, better communication and more immediate report preparation is identified. 6. Individual project evaluation is given for NRA (or the appropriate Water Authority predecessor) tracking studies conducted on the Ribble estuary, the River Tamar, River Torridge, Rivers Test and Itchen, River Lodden, the Welsh River Dee, River Glaslyn, River Taff, River Tawe, River Tywi, River Usk, Rivers Avon and Stour and the River Frome. 7. An outline for future strategic research is provided which identifies particular areas for study:- i) Identification of environmental factors which control the entry of fish into rivers. ii) Improvement of the understanding of the relationship between water flow and upstream movement of salmonids. iii) Examination of the detailed movements and behaviour of fish in relation to obstructions. iv) Closer definition of water quality requirements for salmonid fish. v) Definition of habitat preferences of salmonids in rivers. vi) Subsidiary topics such as the movements of non-salmonid fish and the downstream migration of kelts and juvenile salmonids.

Influência de um extrato aquoso de Coriandrum sativum na marcação in vitro de constituintes sanguíneos com tecnécio-99m e de sua associação com vibração gerada por plataforma oscilante na biodistribuição do radiofármaco Na99mTcO4 e na concentração de biomarcadores em ratos Wistar

Coriandrum sativum, conhecido popularmente como coentro, é um vegetal usado na alimentação humana. Também é utilizado como planta medicinal para tratamento de diabetes, complicações gastrintestinais, e como um antiedêmico, antisséptico e emenagogo. Em investigações acerca dos efeitos do extrato de plantas, é importante a determinação de alguns parâmetros físico-químicos. Diversos modelos experimentais têm sido usados, inclusive com o emprego de radionuclídeos. Em procedimentos da Medicina Nuclear que auxiliam o diagnóstico de doenças, o tecnécio-99m (99mTc) é o radionuclídeo mais utilizado. Hemácias marcadas com 99mTc estão entre as várias estruturas celulares que podem ser marcadas com este radionuclídeo e usadas como radiofármaco. Para a marcação com 99mTc é necessária a presença de um agente redutor, e o mais utilizado é o cloreto estanoso (SnCl2). As terapias com drogas e condições de dieta além de doenças podem alterar a marcação de constituintes sanguíneos, bem como a biodistribuição de diferentes radiofármacos. A exposição às vibrações geradas por plataforma oscilatória produz exercícios de corpo inteiro. O objetivo deste estudo foi caracterizar a preparação de um extrato do Coriandrum sativum, através de parâmetros físico-químicos, verificar os efeitos desse produto natural na radiomarcação de constituintes sanguíneos e em associação à vibração gerada pela plataforma na biodistribuição de Na99mTcO4 e na concentração de alguns biomarcadores. O extrato de coentro teve a o pico de absorbância em 480 nm. O extrato de coentro foi inversamente correlacionado com a concentração na condutividade elétrica. Foi encontrado o maior valor de pH na menor concentração do extrato (0,5 mg/mL). Não houve uma alteração significativa na marcação de constituintes sanguíneos com 99mTc. E a associação do extrato de coentro e vibração gerada por plataforma com frequência de 12 Hz teve efeito no baço, como observado na fixação do radiofármaco nesse órgão e ação em alguns órgãos alternando a concentração de alguns biomarcadores. Em conclusão, parâmetros físico-químicos podem ser úteis para caracterizar o extrato estudado. Provavelmente, as propriedades redox associadas com substâncias desse extrato podem ser os responsáveis pela ausência do efeito na radiomarcação de constituintes sanguíneos. A determinação da captação do Na99mTcO4 em diferentes órgãos permite verificar que o extrato de coentro sozinho não foi capaz de interferir na biodistribuição do radiofármaco. Contudo o tratamento de animais com vibração gerada pela plataforma alterou significativamente a fixação do pertecnetato de sódio no baço e a concentração do colesterol, triglicerídeo, CK e bilirrubina.

Capacidade de invasão, persistência intracelular e atividade citotóxica de cepas de Aeromonas spp. em modelo celular in vitro e ex vivo

As Aeromonas são consideradas patógenos em potenciais para o homem e animais e estão amplamente distribuídas no ambiente sendo a água e os alimentos importantes veículos de transmissão. Muitos estudos têm demonstrado que a patologia causada pela infecção por Aeromonas é complexa e envolvem inúmeros fatores de virulência, dentre eles a aderência, invasão, enterotoxinas, hemolisinas, exoenzimas, sideróforos, flagelos, formação de biofilme e mecanismos de secreção. No presente estudo, analisamos os mecanismos de patogênese mediados por A. caviae e A. hydrophila, avaliando a participação desses microrganismos nos processos de adesão, invasão, persistência intracelular e citotoxidade celular. Foram utilizados ensaios quantitativos in vitro para testar associação, invasão e persistência intracelular em linhagens celulares HEp-2 e/ou T84. A interação de tecidos intestinais de coelho cultivados in vitro (IVOC) com três cepas de A. caviae originárias de fezes diarréicas também foi avaliada. Observamos que 10 (62,5%) das 16 cepas de Aeromonas spp. de diferentes origens, submetidas aos testes de invasão quantitativos foram capazes de invadir células HEp-2 e T84 em 6 horas de incubação. As cepas positivas nos testes de invasão foram submetidas ao teste quantitativo de persistência em células HEp-2 e sobreviveram no ambiente intracelular por 48 e/ou 72 horas sem multiplicação. A interação de três cepas de A. caviae com a mucosa intestinal de coelho ex vivo resultou em aderência, produção de muco e alterações como, intensa vacuolização e drástica desorganização estrutural que levaram a destruição das microvilosidades intestinais. Este estudo demonstrou que subconjuntos de cepas de A. caviae e A. hydrophila de diversas origens, foram capazes de invadir, persistir ou destruir linhagens celulares in vitro. Nosso estudo também evidenciou que cepas de A. caviae causaram expressivas alterações morfológicas que resultaram na destruição de epitélios intestinais de coelho ex vivo. Finalmente, nossos resultados contribuíram para reforçar o potencial patogênico de cepas de Aeromonas, em especial, as de origem vegetal e clínica.

Estudo das características fenotípicas e genotípicas de amostras de Corynebacterium pseudotuberculosis

Uma vez que C. pseudotuberculosis é o agente etiológico de processos infecciosos em animais caprinos e ovinos e que também pode ser isolado de processos infecciosos em seres humanos as investigações direcionadas para a espécie em questão são necessárias, visto que a escassez de dados epidemiológicos e de conhecimento relativo ao comportamento do microrganismo em hospedeiros animais e humanos em nosso país dificulta o diagnóstico laboratorial da espécie, à semelhança do observado com outra espécie de transmissão zoonótica, o C. ulcerans. Uma preocupação adicional é o fato da espécie em questão também ser capaz de albergar bacteriófagos codificadores da toxina diftérica, representando uma ameaça à circulação dos bacteriófagos. Assim, o presente estudo tem como objetivo geral analisar as características fenotípicas e genotípicas de amostras de C. pseudotuberculosis. Neste sentido, foram propostos os seguintes objetivos: Avaliar as características bioquímicas das amostras através de testes bioquímicos convencionais; avaliar as características bioquímicas das amostras utilizando o sistema semi-automatizado API Coryne; diferenciar amostras de C. pseudotuberculosis de C. ulcerans utilizando a técnica de PCR multiplex; pesquisar a presença de gene tox. Os resultados demonstraram que amostras de C. diphtheriae, C. ulcerans e C. pseudotuberculosis podem ser caracterizadas por métodos bioquímicos convencionais e por taxonomia numérica (API Coryne System). C. ulcerans e C. pseudotuberculosis, com potencial de circulação zoonótica, da mesma forma que C. diphtheriae são capazes de albergar o gene da toxina diftérica. A reação m-PCR foi capaz de discernir as amostras de C. diphtheriae, C. ulcerans e C. pseudotuberculosis e ainda definir o potencial das amostras em produzir a toxina diftérica. Os dados enfatizam a necessidade da técnica multiplex PCR para o diagnostico e para o controle de espécies associadas a quadros de difteria em populações humana.

Avaliação da eficácia de uma nova formulação de anfotericina B poliagregada em diferentes esquemas terapêuticos no tratamento da esporotricose experimental causada por Sporothrix brasiliensis

A esporotricose é uma micose subcutânea, crônica, causada por espécies termo-dimórficas do complexo Sporothrix schenckii. Esta micose apresenta diferentes manifestações clínicas sendo mais comum a forma linfocutânea. Casos graves causados por Sporothrix brasiliensis têm sido descritos recentemente, exigindo um tratamento prolongado com antifúngicos de alta toxicidade como a anfotericina B-desoxicolato ou suas versões menos tóxicas, mas de alto custo. Neste trabalho visamos testar in vitro e in vivo a eficácia de uma nova formulação intravenosa de anfotericina B poliagregada (P-AmB) e testar in vivo sua versão semi-sólida (AmB tópica), comparando-a com o itraconazol (ITC) e a anfotericina B-desoxicolato (D-AmB). Ensaios de susceptibilidade in vitro com S. brasiliensis mostraram que esta espécie é suscetível aos antifúngicos testados. Para os testes de eficácia in vivo foram estabelecidos um modelo de esporotricose disseminada e outro de esporotricose subcutânea, causados por S. brasiliensis. No modelo de esporotricose disseminada camundongos BALB/c foram inoculados intravenosamente com leveduras de S. brasiliensis e, 72 h pós-infecção, tratados sob diferentes regimes terapêuticos: i) uma monoterapia de ITC, D-AmB ou P-AmB; ii) uma combinação terapêutica entre D-AmB e ITC ou P-AmB e ITC; iii) um regime de pulso com D-AmB ou P-AmB. A sobrevivência (n= nove) e a carga fúngica em órgãos internos (n= três, no mínimo) foram avaliadas, sendo observado que o regime de pulso com D-AmB ou P-AmB foi o mais efetivo em prolongar a sobrevivência dos animais e reduzir a carga fúngica nos órgãos, seguido pela combinação terapêutica, porém o tratamento com D-AmB e ITC foi a combinação mais efetiva. A monoterapia com ITC e P-AmB e D-AmB foram menos eficazes, sendo corroborados pelas análises histopatológicas. Ensaios de toxicidade in vivo com as diferentes drogas revelaram que ITC e D-AmB induziram a uma toxicidade hepática e renal nos animais, respectivamente, mas P-AmB não induziu a nenhuma toxicidade. Nos ensaios de citoxicidade in vitro foi observado que ITC foi a menos citotóxica e hemolítica e a mais seletiva das drogas testadas, seguida por P-AmB, que foi menos citotóxica e mais seletiva que D-AmB. No modelo de esporotricose subcutânea camundongos da mesma linhagem foram inoculados por via subcutânea com conídios de S. schenckii e de S. brasiliensis (n=9/ grupo). Os animais infectados com S. brasiliensis apresentaram regressão das lesões primárias e disseminação. Usando o modelo de esporotricose subcutânea murina causada por S. brasiliensis testamos peliminarmente a formulação tópica de AmB poliagregada, que reduziu a extensão das lesões de animais infectados. Este é o primeiro trabalho a avaliar diferentes regimes de tratamento da esporotricose disseminada murina causada por S. brasiliensis utilizando ITC, D-AmB e uma nova formulação menos tóxica de anfotericina B poliagregada. O estudo revelou que o regime de pulso foi o mais eficaz para as formulações intravenosas de AmB. Nosso estudo também estabeleceu pioneiramente um modelo de esporotricose subcutânea induzido por S. brasiliensis, que se revelou uma ferramenta útil para comparar a virulência das espécies do complexo S. schenckii e para testar a eficácia de antifúngicos contra essas novas espécies.

Value chain analysis: shrimp, prawn and tilapia from the southern region of Bangladesh & feasibility analysis: brackish water sea-bass in the southern region in Bangladesh

The WorldFish Center is implementing the FtF Aquaculture Project in 20 southern districts in Bangladesh. The project is implemented under USAID’s Feed the Future initiative in collaboration with the Government of Bangladesh. The project contributes to achieving the ‘Feed the Future’ goals through four objectives: (i) dissemination of improved quality fish and shrimp seed, (ii) improving the nutrition and income status of farm households, (iii) increasing investment, employment and fish production through commercial aquaculture and (iv) policy and regulatory reform and institutional capacity building to support sustainable aquaculture growth. The project commissioned this study to gather insights into the value chains of shrimp, prawn and tilapia in the project region and the feasibility of promoting culture of brackish water sea-bass in the region. The findings and recommendations are expected to provide the foundation for the project to design its interventions for achieving its goals.

高温和光照对光系统I结构与功能的影响

光合膜上包含有捕光并将光能转化为化学能所必需的四类跨膜蛋白复合体，即PSII、PSI、Cytb6f和ATP合成酶，其中PSI利用吸收的光能诱导电子从膜内侧的PC传递至相对一侧的铁氧还蛋白，被还原的铁氧还蛋白在FNR（Fd-NADP+氧化还原酶）的作用下生成NADPH，因此关于PSI的研究是光合作用研究领域中的重大问题。为了进一步阐明PSI的结构和功能，本论文分别研究了热对PSI的影响和光诱导的PSI核心复合物（CPI）的积累过程： 1.以菠菜PSI颗粒为材料研究了热处理对PSI复合物的降解和失活作用; 2.以衣藻叶绿素暗合成突变体y-1为材料研究了类囊体膜形成过程（即光诱导的转绿过程）中PSI中CPI的变化。另外，由于膜脂在光合作用中具有重要的功能，本论文还研究了y-1突变体转绿过程中光合膜脂、脂肪酸的变化。 一．应用光谱学、氧电极和变性电泳等技术研究了高温(25oC~80oC)对PSI结构和功能的影响，主要结果如下： 1． 在热处理过程中，683nm组分（主要归属于LHCI）的吸收峰强度有显著的下降并发生峰位蓝移现象，显示该组分对热处理最敏感，首先遭到破坏。 2． 77K荧光显示随着处理温度的升高，728 nm处的峰强和峰位均发生了明显的变化，F728-F720和F680的比率下降，说明热处理抑制了LHCI 680向LHCI 730以及反应中心的能量传递。 3． SDS-PAGE显示PSI核心蛋白PsaA/B亚基以及LHCI亚基在热处理情况下发生了不同程度的降解和聚合。为了能够显著地观察到热处理对PSI多肽降解的影响，实验采用了更高的温度处理方法，结果显示，90oC、100oC时PsaA/B亚基完全降解，而LHCI亚基仍有少量存在，说明PSI核心蛋白PsaA/B比LHCI亚基具有更高的热敏感性。 4． 推测热处理情况下可能发生的机制是，捕光天线首先从PSI反应中心分离，随后发生了反应中心光化学反应的抑制，直至最后多肽的严重降解。 5． 利用红外光谱技术（FT-IR）对PSI蛋白二级结构的研究显示，PSI颗粒在60oC以上时发生了明显的蛋白构象变化,且随着温度的升高蛋白构象的变化越来越大，表明PSI蛋白具有较高的热稳定性和热变性温度，PSI蛋白酰胺I带(1700~1600 cm-1)二级结构的解析表明热处理过程中二级结构的主要变化是α-helical的下降和β-sheet的增加。 6． 利用CD光谱技术研究了热处理对PSI色素微环境的影响，结果表明热处理破坏了PSI色素蛋白复合物中色素的蛋白微环境，归属于LHCI的Chlb（645 nm处组分）在较低的温度处理条件下(25~60oC)蛋白微环境即发生破坏;随着处理温度的升高（70和80 oC），478 nm处（主要归属于LHCI的Chlb）和498 nm（归属于类胡萝卜素）处的CD信号强度快速下降，说明在高温条件下LHCI比核心复合物更敏感。 7． 研究发现，PSI的摄氧活性随着处理温度的升高而显著下降，70oC时几乎完全失去摄氧能力，表明70oC时PSI复合物受到了严重的破坏，这可能是由于热处理过程中色素的蛋白微环境以及蛋白结构尤其是PSI核心蛋白PsaA/B中跨膜α-helix的构象发生了严重的变化。 二．主要运用温和电泳和蛋白印迹技术检测了暗培养4天(脱绿)的y-1突变体在光照诱发的转绿过程中PSI的核心色素蛋白复合物（CPI）及其叶绿素脱辅基蛋白PsaA/B的变化。 1． 暗培养4天的衣藻脱绿细胞中，PSI的主要色素蛋白复合物-CPI完全缺失，然而核心多肽PsaA/B仍有一定量的积累，同时检测不到P700的含量。 2． 当脱绿的y-1细胞转移至光照下时，伴随着叶绿素的合成，色素蛋白复合物CPI和PsaA/B脱辅基蛋白的合成也逐渐达到正常水平，说明叶绿素和PsaA/B蛋白进行组装并形成了具有功能的PSI反应中心,P700含量也得到了恢复。 3． 实验证明了光照是形成光合系统色素蛋白复合物的重要前提。同时发现，叶绿体基因编码的PSI核心多肽PsaA/B能够在暗条件下合成。而根据资料(Berends et al.,1987)报道，在豌豆、大麦的黄化体中不能合成PsaA/B蛋白，这可能是由于在脱绿的y-1细胞中叶绿体仍然具有相对完整的大小和形状，而在叶绿体的被膜上定位着多种与光合作用相关的酶系统。 三. 利用薄层层析及气相色谱分析技术对转绿期间y-1突变体光合膜脂和脂肪酸组成的含量变化进行了分析。结果表明： 1． 光照能够促进各种脂的积累并影响脂的组成，同时有利于脂肪酸脱饱和酶的激活。MGDG中脂肪酸不饱和程度明显升高，表现为16:0及18:1的下降，以及16:4，18：2和18:3（9，12，15）等的升高，说明光照促进了MGDG sn-2位的16:0脱饱和为16:4以及sn-1位的18：1脱饱和为18:3，也说明MGDG是脂肪酸脱饱和的重要底物。 2． 已有的资料（Ohnishi和Yamada，1980;1983）指出PG及其sn-2位的16:1（3t）的合成是光依赖的，而本实验中，在衣藻的黄化细胞中含有相当量的PG及其特有的16:1（3t）（22.80%），且转绿过程中变化并不十分显著。这可能是由于黄化的y-1细胞中叶绿体的形状并没有发生很大变化，说明叶绿体被膜上的相应酶系统才是PG合成的必需因素。 3． 相比于脱绿的y-1细胞，光照12小时后，各种脂中18：2的百分含量有显著的增加，这来源于18：2是脂肪酸脱饱和的重要中间产物。

甘油和蔗糖对光系统I结构与功能的影响

在光系统I(PSI)内部结合有大量的水分子，而这些水分子的生理功能还不清楚。在本工作中，我们通过外加具有渗透活性和吸水强的多羟基化合物甘油和蔗糖来改变PSI环境中水的含量，研究水的改变对PSI结构与功能的影响。主要结果如下： 1．甘油和蔗糖对PSI的电子传递产生影响，影响程度和大小与它们的浓度有关。一般地，低浓度的甘油和蔗糖可促进PSI的电子传递，而在高浓度时，这种促进作用有所减弱。但过高浓度的甘油(>60%, v/v)会抑制PSI的电子传递活性。 2．与对PSI电子传递的影响趋势相类似，在低浓度的甘油和蔗糖存在下，PSI的光化学反应活性(PSI反应中心色素P700的光氧化还原能力)大为增加，而较高浓度的甘油和蔗糖对P700的氧化还原能力有所抑制。 3．甘油和蔗糖也会改变PSI中的主体色素(bulk chlorophyll)和长波色素或红色素(red chlorophyll)之间的能量分布。它们的作用导致激发能分配失衡，使更多的激发能分配到红色素。 4．甘油和蔗糖的作用还会影响PSI的蛋白质构象。甘油使PSI蛋白质内部的色氨酸残基(Trpapolar)处于更加疏水的微环境，而蔗糖却使极性环境中的色氨酸残基(Trppolar)周围微环境的极性继续增大。它们均会使色氨酸残基邻近的具有淬灭活性的蛋白质的位置和/或方向有所变化。同时，甘油和蔗糖的作用也会导致PSI的疏水性增加。