Intermédiaires réactionnels de la fructose-1,6-biphosphate aldolase de Leishmania mexicana : comparaison avec l'enzyme de mammifère et recrutement de la région C-terminale

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Intermédiaires réactionnels de la fructose-1,6-biphosphate aldolase de Leishmania mexicana : comparaison avec l'enzyme de mammifère et recrutement de la région C-terminale

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Acetilação do exopolissacarídeo (1→6)-β-D-glucana (lasiodiplodana): derivatização química e caracterização

Lasiodiplodan is an exocellular β-glucan with biological functionalities such as antioxidant, antiproliferative, hypocholesterolemic, protective activity against DNA damage induced by doxorubicin and hypoglycemic activity. Chemical derivatization of polysaccharide macromolecules has been considered as a potentiating mechanism for bioactivity. In this context, this work proposes the derivatization of lasiodiplodan by acetylation. Acetic anhydride was used as derivatizing agent and pyridine as catalyst and reaction medium. The derivatives obtained were evaluated by its water solubility, degree of substitution (DS), antioxidant potential, and characterized by infrared spectroscopy (FT-IR), thermal analysis, differential scanning calorimetry, X-ray diffraction and scanning electron microscopy. Acetylated derivatives with different degrees of substitution (1.26; 1.03; 0.66 and 0.48) were obtained, and there was correlation between the concentration of derivatizing agent and DS. FT-IR spectroscopy analysis confirmed the insertion of acetyl groups into derivatized macromolecules (LAS-AC) through of specific bands concerning to carbonyl group (C = O) and increase in C-O vibration. SEM analysis indicated that native lasiodiplodan presents morphological structure in the form of thin films with translucent appearance and folds along its length. Derivatization led to morphological changes in the polymer, including aspects thickness, translucency and agglomeration. Thermal analysis indicated the native sample and derivative with DS 0.48 presented three weight loss stages. The first stage occurred until 125 ° C (loss of water) and there were two consecutive events of weight loss (200 ° C - 400 ° C) attributed to molecule degradation. Samples with DS 1.26; 1.03 and 0.66 demonstrated four weight loss stages. The first stage occurred until 130 ° C (loss of water), following by two consecutive events of weight loss (200 ° C - 392 ° C) attributed to degradation of the biopolymer. The fourth stage was between 381 ° C and 532 ° C (final decomposition) with exothermic peaks between 472 ° C and 491 ° C. X-ray diffraction patterns showed that native and acetylated lasiodiplodan have amorphous structure with semicrystalline regions. Derivatization did not contribute to increased solubility of the macromolecule, but potentiated its antioxidant capacity. Acetylation of lasiodiplodan allowed to obtaining a new macromolecule with higher antioxidant potential than the native molecule and with technological properties applicable in various industrial sectors.

大口胭脂鱼线粒体DNA控制区序列的研究

采用PCR扩增、直接测序的方法得到了大口胭脂鱼线粒体DNA控制区的序列 ,经过对比分析 ,其序列全长为 92 0bp,碱基含量分别为 :胸腺嘧啶 (T) 2 9.8% ,胞嘧啶 (C) 2 1 .6% ,腺嘌呤 (A) 3 2 .4 % ,鸟嘌呤 (G)1 6.2 % ,与其它鲤科鱼类相比 ,胸腺嘧啶含量略低 ,鸟嘌呤含量略高 ,其它则相似 .成功地识别了终止序列区(nt.1～ 2 3 5处 )、中央保守区 (nt2 3 6～ 5 66处 )和保守序列区 (nt.5 67～ 92 0处 ) ,并找到了终止

原子吸收光谱法提高某些元素灵敏度的方法研究及应用

原子吸收光谱法自五十年代提出以来发展十分迅速，迄今已是一种比较成熟的仪器分析方法。对大多数元素的研究已趋于完善，要提高分析灵敏度比较困难。本工作主要从基体改进剂的角度来研究石墨炉原子吸收光谱法（GFAAS）提高某些元素，尤其是高温元素的测定灵敏度的方法及其应用。GFAAS测硼的报道很少，硼的分析灵敏度低原因的于硼在低温时易以氧化物形式挥发损失，而高温时又生成难熔的B_4C。因此，必须寻找合适的基体改进剂。本工作通过对基体改进剂量的试验以及基体改进剂对硼的灰化和原子化温度影响的研究，得到3Ca, Sr, Ca/Mg、Ca/La和Sr/Mg均可作为测硼的基体改进剂的结果，其中混合基体改进剂Ca/Mg、Ca/La和Sr/Mg较Ca、Sr单独作基体改进剂性能要优越。本工作将x射线衍射分析与热力学计算相结合，综合已有的实验现象，提出了硼原子化过程的初步机理，同时也解释了基体改进剂Sr(NO_3)_2对硼吸收信号的增强作用，并推测了相应原子化过程。即气态硼原子的产生源自固态硼原子的升华，而固态硼原子的产生源自B_2O_3的碳还原。Sr(NO_3)_2的增感作用在于子化前SrB_6的生成，减少了B_2O_3的挥发损失，阻止了B_4C的生成。此外，本工作在基体改进剂研究的基础上，选用Ca/Mg和Sr/Mg混合基体改进剂分别测定了饮用水，标准参考物质桃叶以及高温镍基合金钢中的痕量硼，进行了灵敏度、线性范围和抗干扰能力试验。硼1％吸收时的特征量分别为2.5 * 10~(-10)g和2.0 * 10~(-10)g，线性范围均达1.6μg ml~(-1)，回收率在98-105%之间，精密度为4.7-8.4%。饮用水的检出限为50ng ml~(-1) B(3σ)。本工作还比较了金属盐类注入和浸渍处理石墨管后，GFAAS测硼的灵敏度、精密度和石墨管的使用寿命。用扫描电镜分析了石墨管的表面结构，并作了相应解释。建立快速、准确和灵敏的方法检测环境试样中的痕量Be、Al、Sn和Ge，是环境分析的重要课题之一。GFAAS作为一种灵敏的分析方法已广泛地用于环境分析上，但测定某些试样中的Be、Al、Sn和Ge灵敏度尚低。为此，本工作以Ca(NO_3)_2为基体改进剂探讨了增强这四种元素灵敏度的方法。实验结果表明，最佳条件下对比测定，Ca(NO_3)_2能将Be、Al、Sn和Ge的分析灵敏度分别提高3.6，5.7，9和50倍。Ca(NO_3)_2不仅提高了Be、Al、Sn和Ge的分析灵敏度，而且也提高灰化温度，并降低原子化温度，也增强抗干扰能力。本工作还探讨了Ca(NO_3)_2对Be、Al、Sn和Ge的增感机理。实验结果表明，钙对Be和Ge的增感主要在于气相作用，钙对Al的增感主要是固相作用，而钙对Sn的增感不仅有气相作用，也有固相作用。同时利用钙的增感作用直接测定了地下水及水系沉积物标样中的痕量铍，其检出限为5.5 * 10~(-13)g Be(3σ)，回收率在98-105%之间，精密度为2.8-4.4%。也利用钙的增感作用直接测定了河水及地下水中的痕量铝，其检出限为1.1ng ml~(-1) Al (3σ)，回收率在95-106%之间，测定的精密度为2.7-6.8%。此外，本工作也研究了石墨炉原子吸收光谱法测铝时各种氯化物的干扰现象。结果发现，NaCl，KCl，FeCl_3和CuCl_2对铝产生负干扰，这种干扰来自气相中Cl和Al的结合；MgCl_2和CaCl_2对铝产生正干扰，这种干扰来自它们的氧化物对铝的增感作用；HCl不干扰铝的测定，而HClO_4对铝的干扰与石墨管的表面有关。本工作所建立的GFAAS测Al，B，Be的方法，具有简单、快速，且灵敏度高的特点，适用于环境试样中痕量铝、硼、铍的测定。

TCAS树脂的制备及对水中污染物的吸附性能研究

本论文采用第三代超分子受体化合物——磺化硫杂杯芳烃（简称TCAS）作为活性组分，将其加载到树脂载体上，制成一种新型的TCAS吸附树脂。借助红外光谱、紫外光谱、核磁共振和电喷雾质谱等检测分析手段对TCAS吸附树脂及其制备的中间产物进行表征。采用静态批试验方法研究发现TCAS吸附树脂对重金属和卤代烃类有机物都有较好的吸附去除效果，其对Pb2+、Cd2+、Cu2+和Zn2+四种重金属的吸附容量分别达到26.32mg•g-1、18.12mg•g-1、12.24mg•g-1和6.85mg•g-1；对二氯甲烷、三氯甲烷、四氯化碳、三氯乙烯四种卤代烃的吸附容量分别为9.23 mg•g-1、7.92 mg•g-1、6.73 mg•g-1和4.34 mg•g-1。并发现温度、树脂用量和溶液pH值等因素能影响TCAS吸附树脂的吸附去除效果。同时研究了TCAS吸附树脂对污染物的吸附优先性规律、动力学规律和热力学规律。采用化学计量学分析TCAS络合重金属的紫外光谱，获得了TCAS－重金属络合物的纯紫外光谱，以及TCAS、TCAS－重金属络合物随重金属浓度变化的分布曲线，并最终解析得到TCAS与Cu、Zn、Cd和Pb的络合常数（lgβ）分别为9.79、8.72、6.87和5.00。通过多次吸附试验，考察了TCAS吸附树脂回收循环利用的可行性，发现TCAS吸附树脂可进行多次循环回收再利用。

阿哈水库中不同形态汞迁移转化规律的初步研究

阿哈水库是贵阳市集中式生活饮用水源地一级保护区，具有极为重要的社会和经济功能。长久以来阿哈水库流域内乡镇煤矿、个体煤窑矿采严重，大量酸性矿山废水未经处理直接入库以及近几年来的人工石灰投放使阿哈水库水体成为罕见的高铁锰含量的硫酸盐型水体。本研究于同一水文年的枯水期（2005年3月）和丰水期（2005年8月）分别对水库不同深度分层水体、沉积物间隙水、沉积柱剖面、阿哈水库所有支流水体中不同形态汞的含量、分布特征以及迁移转化规律进行了详细的研究。研究结果发现： 1. 阿哈水库水体中各种形态汞浓度均出现季节性变化。总汞(HgT)浓度变化范围为2.1~19.5 ng•L-1，平均为9.3ng•L-1；溶解态汞(HgD)浓度变化范围为1.7~ 9.0 ng•L-1，平均为4.1ng•L-1；颗粒态汞(HgP)浓度范围为0.3~14.8 ng•L-1，平均5.2 ng•L-1；活性汞(HgR)浓度范围为0.1~ 4.5 ng•L-1，平均1.7 ng•L-1； 2. 阿哈水库水体颗粒态汞与总汞呈极显著相关，说明阿哈水库水体的颗粒态汞强烈影响着总汞迁移和分布。水库水体甲基汞与溶解态汞之间存在极显著正线性相关，证明水库水体中的甲基汞的产生和迁移与溶解态汞有着密切的关系，但是，水体甲基汞与总汞之间并不存在相关性。 3.阿哈水库水体丰水期溶解态汞、活性汞、溶解态甲基汞浓度均远高于枯水期浓度，除了外源径流输入的影响外，很大程度与丰水期水库底部缺氧，氧化还原界面由沉积物-水界面进入水体密不可分。较大表面积铁锰氧化物还原为可溶态的铁锰离子，使沉积物释放大量被吸附的汞进入水体，造成水体溶解态汞、活性汞、溶解态甲基汞的升高。 4. 阿哈水库沉积物间隙水中总汞和甲基汞浓度出现明显的季节性变化，丰水期〉枯水期。间隙水中甲基汞浓度与沉积物中甲基汞含量呈极显著性相关。阿哈水库沉积物间隙水中甲基汞的浓度除了沉积物表层0cm到表层下8cm出现峰值外，在表层10cm以下仍然发现有浓度波动，两者可能归因于汞的甲基化进程受硫酸盐还原菌和铁还原菌的双重影响。 5. 阿哈水库沉积物总汞含量在枯水期和丰水期基本没有变化，变化范围为160-252 ng•g-1，平均为209 ng•g-1，大大高于其他未受污染地区水库沉积物中总汞含量。甲基汞含量变化范围为0.22-7.21 ng•g-1，平均为1.76 ng•g-1。 6. 阿哈水库入库河流pH为7.5-8.5，总汞浓度为7.1-47.9 ng•L-1；甲基汞浓度为0.16-2.50 ng•L-1；活性汞浓度为0.25-2.43ng•L-1，且随季节变化明显，枯水期<丰水期。阿哈水库入库河流汞的主要存在形态为颗粒态汞，总汞与颗粒态汞呈显著正相关。九十年代末期至今，矿山煤窑废水经集中治理，阿哈水库流域的煤窑废水污染状况得到很大改善。目前，阿哈水库汞的主要来源为未经处理直接排入水库中的生活污水和其它工业废水。阿哈水库唯一的出湖河流－小车河中，各种形态的汞均较入湖河流中的汞含量大大减少，这表明水库的沉淀和拦截净化作用相当显著。

Pharmacological studies of 8-OH-DPAT-induced pupillary dilation in anesthetized rats

Serotonin (5-HT) receptor agonists have been reported to produce mydriasis in mice, and miosis in rabbits and humans. However, the underlying mechanisms for this action are unclear. This study was undertaken in an attempt to explore the mechanism by which 5-HT receptors are involved in the modulation of pupillary size in pentobarbital-anesthetized rats. Intravenous administration of the 5-HT receptor agonist, (2R)-(+)-8-hydroxy-2-(di-n-propylamino)tetralin hydrobromide (8-OH-DPAT; 0.003-3 mg/kg), elicited dose-dependent pupillary dilation, which was not affected by section of the preganglionic cervical sympathetic nerve. 8-OH-DPAT-elicited mydriatic responses were attenuated by the selective 5-HT receptor antagonist, N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-N-2- pyridinylcyclohexanecarboxamide maleate (WAY 100635; 0.3-1 mg/kg, i.v.), as well as by the selective a -adrenoceptor antagonist, (8aR,12aS,13aS)-5,8,8a,9,10,11,12,12a,13,13a-dechydro-3-methoxy-12- (ethylsulfonyl)-6H-isoquino[2,1-g][1,6]naphthyridine hydrochloride (RS 79948; 0.3 mg/kg, i.v.), but not by the selective a -adrenoceptor antagonist, prazosin (0.3 mg/kg, i.v.). Mydriatic responses elicited by the a -adrenoceptor agonist, guanabenz (0.003-0.3 mg/kg, i.v.), were not antagonized by WAY 100635 (0.3-1 mg/kg, i.v.). To determine whether central nervous system (CNS) 5-HT receptors, like a -adrenoceptors, are involved in reflex mydriasis, voltage response curves of pupillary dilation were constructed by stimulation of the sciatic nerve in anesthetized rats. WAY 100635 (1 mg/kg, i.v.) did not antagonize the evoked reflex mydriasis, which, however, was blocked by RS 79948 (0.3 mg/kg, i.v.). Taken together, these results suggest that 8-OH-DPAT produces pupillary dilation in anesthetized rats by stimulating CNS 5-HT receptors, which in turn trigger the release of norepinephrine, presumably from the locus coeruleus. The latter reduces parasympathetic neuronal tone to the iris sphincter muscle by stimulation of postsynaptic a - adrenoceptors within the Edinger-Westphal nucleus. Unlike a - adrenoceptors, 5-HT receptors in the CNS do not mediate reflex mydriasis evoked by sciatic nerve stimulation. © 2004 Elsevier B.V. All rights reserved.

Hydrodynamic and sediment fluxes through the inlets of the Ria Formosa

Tese de dout., Ciências do Mar, da Terra e do Ambiente (Ciências do Mar-Oceanografia Física), Faculdade de Ciências e Tecnologia, Univ. do Algarve, 2011

Fluorescent ionene-dye nanoparticles by electrostatic self-assembly

It is investigated that the association of linear cationic model polyelectrolytes with oppositely charged pyrenetetrasulfonate (PY) in aqueous solution. For this purpose water soluble ionenes were prepared via Menschutkin reaction from 1-4-diazabicyclo [2.2.2] octane and e.g. 1,6-dibromohexane and 1,4 dibromotransbuten. The complex formation between dye molecules PY and oppositely charged ionenes (PD4, PD6, PD4-2 and PD4coPD6) of different chemical structures in aqueous solution was studied by light scattering (LS), small angle neutron scattering (SANS), UV-Vis, fluorescence spectroscopy and atomic force microscopy (AFM). Spectrophotometric titration results revealed that PY molecules were bind to ionenes cooperative process due to π-π interaction. Cooperative binding constant KD was determined as 6.4 x 10^6 M^-1 (+ or - 10^5 M^-1). It was found that binding mode and geometry of PY is predominantly depending on inter-charge distances of corresponding ionenes. Resultant complexes have exhibited size and structure variation as a function of charge ratio (L), ionic strength, inter-charge distances. Spherical dye-ionene complexes of which radius of gyration ranging between (RG) 50 and 190 nm have been observed in PD4-PY system while this was not possible with a different ionene (PD6) or either case ionene excess. It was found that most of the PD4-PY complexes had RG / RH ~ 0.78. Based on the AFM and LS results, spherical complexes have certain colloidal stability and their size can effectively controlled by changing the L.

Interval training program optimization in highly trained endurance cyclists

PURPOSE: The purpose of this study was to examine the influence of three different high-intensity interval training (HIT) regimens on endurance performance in highly trained endurance athletes. METHODS: Before, and after 2 and 4 wk of training, 38 cyclists and triathletes (mean +/- SD; age = 25 +/- 6 yr; mass = 75 +/- 7 kg; VO(2peak) = 64.5 +/- 5.2 mL x kg(-1) min(-1)) performed: 1) a progressive cycle test to measure peak oxygen consumption (VO(2peak)) and peak aerobic power output (PPO), 2) a time to exhaustion test (T(max)) at their VO(2peak) power output (P(max)), as well as 3) a 40-km time-trial (TT(40)). Subjects were matched and assigned to one of four training groups (G(2), N = 8, 8 x 60% T(max) at P(max), 1:2 work:recovery ratio; G(2), N = 9, 8 x 60% T(max) at P(max), recovery at 65% HR(max); G(3), N = 10, 12 x 30 s at 175% PPO, 4.5-min recovery; G(CON), N = 11). In addition to G(1), G(2), and G(3) performing HIT twice per week, all athletes maintained their regular low-intensity training throughout the experimental period. RESULTS: All HIT groups improved TT(40) performance (+4.4 to +5.8%) and PPO (+3.0 to +6.2%) significantly more than G(CON) (-0.9 to +1.1%; P < 0.05). Furthermore, G(1) (+5.4%) and G(2) (+8.1%) improved their VO(2peak) significantly more than G(CON) (+1.0%; P < 0.05). CONCLUSION: The present study has shown that when HIT incorporates P(max) as the interval intensity and 60% of T(max) as the interval duration, already highly trained cyclists can significantly improve their 40-km time trial performance. Moreover, the present data confirm prior research, in that repeated supramaximal HIT can significantly improve 40-km time trial performance.

Optimization of Off-Null Ellipsometry for Optical Biosensor Applications

The optimization of off-null ellipsometry is described with emphasis on the improvement of resolution for visualizing biomolecule layers. For optical biosensor with layer thickness below 6.5 nm, a numerical simulation for the dependence of resolution on the azimuth settings of polarizer and analyzer is presented first. For comparison, three different resolutions are given at three azimuth settings which are near null and far away from null condition, respectively. Furthermore, the square or linear approximation relationship between the intensity and the layer thickness are also given at these settings. The difference among their accuracy is up to 100 times or so. Experimental results of the biosensor sample verify the optimization.