乌头类双酯型生物碱对嗜热四膜虫生长代谢的微量热学研究

建立灵敏表征乌头类双酯型生物碱(DDA)对嗜热四膜虫生长代谢影响的评价方法. 采用微量热法, 在不同给药条件下, 以表达功率-时间曲线(热谱曲线)的特征参数生长速率常数(k)、半数抑制浓度(IC₅₀)、最大输出功率(P_max)及达峰时间(t_p)和总产热量(Q_t)为指标, 对嗜热四膜虫生长代谢程度进行客观地量化评价. 结果表明, 随着3种DDA浓度的增加, 达峰时间t_p延长, 生长速率常数k、最大输出功率P_max等随之降低, 且生长速率常数k与相应的浓度间呈现良好的线性关系: r_乌头碱＝0.9910 (IC₅₀＝207.7 μg•mL^－1); r_新乌头碱＝0.9923 (IC₅₀＝412.5 μg•mL^－1); r_次乌头碱＝0.9977 (IC₅₀＝1497 μg•mL^－1). 同时由半抑制浓度IC₅₀得到3种DDA毒性的大小顺序: 乌头碱＞新乌头碱＞次乌头碱. 初步构效关系研究表明, 在化合物C(3)上引入羟基(OH)以及N原子上引入乙基(CH₂CH₃)均可明显增加双酯型生物碱对嗜热四膜虫的毒性作用.     

N,N'-二(二乙氧基硫代磷酰基)-1,4-苯二胺的合成、晶体结构及热性能研究

通过二乙氧基硫代磷酰氯与对苯二胺反应生成了标题化合物N,N'-二(二乙氧基硫代磷酰基)-1,4-苯二胺，并应用元素分析, FTIR及¹H NMR对标题化合物进行了表征. 利用X射线单晶衍射测定了其晶体结构, 同时应用TG分析法对其热性能进行了分析. 标题化合物的相对分子质量M_r＝412.42, 为正交晶系, Pbca空间群, 晶胞参数为a＝0.86936(16) nm, b＝1.2787(2) nm, c＝1.8897(3) nm, β＝90°, V＝2.1006(7) nm³, Z＝8, D_c＝1.304 g/cm³, μ(Mo Kα)＝0.425 mm^－1, F(000)＝872, S＝1.052. 最终偏离因子R＝0.0628, wR＝0.1860, 可观测衍射点1852个[I＞2σ(I)]. 该晶体通过对苯二胺连接并以中心对称分布, 并形成层状结构, 且存在弱的分子内氢键N—H…S. TG分析表明该化合物有很好的热稳定性及成炭性, 通过其阻燃聚丙烯腈表明, 该物质是一种高效能的膨胀型阻燃剂.     

α-取代氨基氟代苯基膦酸酯衍生物的合成、晶体结构与抗癌活性

利用席夫碱与亚磷酸酯反应, 合成了新型O,O'-二烷基-α-(6-甲氧苯并噻唑-2-基氨基)-4-氟苯基膦酸酯化合物, 结构经元素分析, IR, ¹H NMR, ¹³C NMR和X单晶衍射确认. X单晶衍射测试结果表明: 化合物3d分子属于四面体晶系, 空间群I4(1)/a, a＝2.1055(3) nm, b＝2.1055(3) nm, c＝2.0521(5) nm, α＝90.00°, β＝90.00°, γ＝90.00°, V＝0.9098(3) nm³, Z＝16, D_c＝1.321 mg/m³, ＝0.250 mm^－1, F(000)＝3808. 化合物还存在着1个分子内氢键[N(2)—H(2)…O(1)]. 生物测定表明化合物3f在20 g/mL浓度下对PC3细胞的抑制率为84.3%.     

[1-芳基甲羰基-2-(1,2,4-三唑-1-基)]乙基-N,N-二烷基二硫代氨基甲酸酯衍生物的合成、结构表征及生物活性研究

以芳香基三唑类杀菌剂三唑酮为先导物设计并合成了5个含N,N-二烷基二硫代氨基甲酸酯的芳香三唑类化合物, 通过元素分析、红外光谱、核磁共振氢谱和质谱对其结构进行了表征. 用X射线单晶衍射测定了[α-(4-甲氧基苯甲酰基)-2-(1,2,4-三唑-1-基)]乙基-N,N-二甲基二硫代氨基甲酸酯的晶体结构, 晶体属于三斜晶系, 空间群, 晶胞参数为: a＝0.73482(15) nm, b＝1.1051(2) nm, c＝1.1209(2) nm, α＝90.32(3)°, β＝101.97(3)°, γ＝105.13(3)°, V＝0.8578(3) nm³, Z＝2, D_c＝1.357 g/cm³, F(000)＝368, µ＝0.324 mm^－1. 生物测试结果显示这5种有机化合物都具有杀菌性和植物生长调节活性     

钇二氢化反应热力学函数的研究

用密度泛函方法B3LYP/SDD/6-311＋＋G**计算了YH₂的微观性质. 并用分子总能量中的振动能E_v代替固态能量0705212, 振动熵S_Ev代替固态熵的近似方法, 以及考虑到电子能量的变化, 计算了固态YH₂(D, T)的焓H和熵S, 得到不同温度下Y与H₂, D₂, T₂反应的ΔH^⊖, ΔS^⊖, ΔG^⊖及氢化反应平衡压力, 导出了与温度的依赖关系. 计算结果表明, YH₂(s)的生成热为199.25 kJ•mol^－1, 与实验值210.00及 225.94 kJ•mol^－1非常接近, 说明近似方法的正确性.     

超声辐射下O,O'-二正丁基-α-(4-三氟甲基苯胺基)-2-氟苯基膦酸酯的合成与晶体结构

采用BF₃•Et₂O催化和45 kHz超声辐射下亚磷酸丁酯与邻氟苯甲醛、对三氟甲基苯胺在78～80 ℃, 无溶剂条件下进行类 Mannich 反应, 0.5 h即可以较高收率获得目标化合物α-氨基膦酸酯(4), 其结构经元素分析, ¹H NMR, IR及MS确认. 对化合物晶体结构进行X衍射分析, 结果表明化合物属单斜晶系, 空间群P2(1)/c, 晶胞参数a＝1.348(4) nm, b＝1.734(4) nm, c＝1.099(3) nm, α＝90°, β＝109.71(4)°, γ＝90°, V＝2.417(11) nm³, Z＝4, D_c＝1.268 g/m³, μ＝0.166 mm^－1, F(000)＝968.     

线性BC2nB (n＝1～12)的结构特征和电子光谱的理论研究

应用密度泛函理论, 在B3LYP/6-31G^*水平上优化得到了线性簇合物BC_2nB (n＝1～12, D_(h)的平衡几何构型, 并计算了它们的谐振动频率. 在优化平衡几何构型下, 通过TD-B3LYP/cc-pvDZ和TD-B3LYP/cc-pvTZ计算, 分别得到了n＝1～12和n＝1～7的电子跃迁的垂直激发能和对应的振子强度. 在B3LYP/6-311＋G^*水平上计算得到了簇合物BC_2nB (n＝1～12, D_(h)的电离能. 基于计算结果, 导出了BC_2nB体系电子跃迁能以及第一电离能与体系大小n的解析表达式.     

(GaP)n和(GaP)n－ (n＝10～16)团簇的结构与稳定性研究

采用密度泛函理论B3LYP/Lanl2dz方法对(GaP)_n和(GaP)_n^－ (n＝10～16)团簇的一系列异构体的结构和稳定性进行了研究. 讨论了中性团簇得到一个电子之后, 几何结构和电子性质的变化. 频率分析预测出最强吸收峰位于341～390 cm^－1区域. 从能隙、结合能和能量二次差分等方面综合考虑, 具有T_h对称性的(GaP)₁₂和(GaP)₁₂^－分别是中性(GaP)_n和阴离子(GaP)_n^－团簇中最稳定的, 而具有T_d点群结构的(GaP)₁₆也比较稳定, 究竟哪种结构易于合成还有待于实验的进一步证实. 在相同理论水平上计算了基态(GaP)_n (n＝10～16)的绝热电子亲合势(AEAs)及其基态阴离子的垂直电离能(VDEs), 这对以后的实验数据分析将有一定的参考价值.     

层状MAgTeS3 (M＝K, Rb)的溶剂热合成与表征

以硫醇为螯合剂, 在溶剂热条件下合成了两种层状硫代亚碲酸盐KAgTeS₃ (1)和RbAgTeS₃ (2). X射线单晶解析表明, 1和2是类质同晶化合物. 在晶体结构中, 银硫四面体通过共用顶点形成无限的平行链, 在相邻链中银硫四面体取向相反, 这些链与链由三角锥配位的碲互相连接形成阴离子层状结构, 阳离子在阴离子层间. 1的结晶学数据为: M_r＝370.75, P2₁/c, a＝0.73639(6) nm, b＝1.06468(8) nm, c＝0.85203(6) nm, β＝106.4640(10)°, V＝0.64062(8) nm³, Z＝4, R(F)＝4.44%, wR(F²)＝11.66%. 2的结晶学数据: M_r＝417.12, P2₁/c, a＝0.75531(12) nm, b＝1.07076(7) nm, c＝0.8583(2) nm, β＝106.497(6)°, V＝0.66558(19) nm³, Z＝4, R(F)＝6.00%, wR(F²)＝15.43%. DSC及紫外-可见漫反射光谱研究表明, 这两种化合物为半导体, 并具有很好的热稳定性.     

HCF(X~1A’)+SO2反应的动力学研究

用激光光解-激光诱导荧光方法研究了室温下(T＝293 K) HCF(X^~1A^’)自由基与SO₂分子的反应动力学. 实验中HCF(X^~1A^’)自由基是由213 nm激光光解HCFBr₂产生的, 用激光诱导荧光(LIF)检测HCF(X^~1A^’)自由基的相对浓度随着反应时间的变化, 得到此反应的二级反应速率常数为: k＝(1.81±0.15)×10^－12 cm³•molecule^－1•s^－1, 体系总压为1862 Pa. 高精度理论计算表明, HCF(X^~1A^’)和SO₂分子反应的机理是典型的加成-消除反应. 我们运用RRKM-TST理论计算了此二级反应速率常数的温度效应和压力效应, 计算结果和室温下测定的二级反应速率常数符合得较好.     

Study on eruption of heat for Escherichia coli B aroused by lanthanum nitrate and its mechanism

Biological effect of rare-earth lanthanum nitrate on the growth of Escherichia coli B was studied using the calorimetric method. There were exceptional changes on the growth thermogenic curves for high concentrations of lanthanum nitrate. For example, the peak high, the total quantity of heat (Q) of cultures and the growth rate constants (k) are evidently increased when compared with normal E. coli B cultures. When the concentration of lanthanum nitrate was at 300 mg/L and 500 mg/L, and Q of the cultures reached 3.89 and 2.54 times of normal cultures, respectively. The survivability of cells and the biomass of the cultures were measured using biological methods and the results show that the growth and multiplication of cells were inhibited and that the biomass decreased at high concentration of lanthanum nitrate. These revealed that the inhibiting cells discharged more quantity of heat than the normal growing cells. We named this phenomenon as “eruption of heat”. It was suggested that the mechanism for the eruption of heat was that La³⁺ ion damages the outer cell membrane and increases its permeability and the proton-electron potential energy across the cell membrane was reduced or couldn’t even be initiated. Energy could not be translated into ATP effectively in the course of oxidative phosphorylation resulting in heat release. So, the growth of the cells was inhibited due to scarceness of energy ATP. __________ Translated from Acta Chimica Sinica, 2007, 65(10): 917–922 [译自: 化学学报]     

鞘磷脂和神经节苷脂对磷脂酰乙醇胺混合脂超分子体系液晶态立方相Im3m和Pn3m结构影响的研究

用磷脂酰乙醇胺(DEPE)、鞘磷脂(Sphingomyeline, Sph)、神经节苷脂(Gm1)和胆固醇(Chol)模拟了生物膜超分子体系液晶态结构, 通过用小角X射线衍射(SAXD)对混合脂体系液晶态结构进行了研究, 鉴定出了两种立方相: 即Im3m(Q²²⁹)和Pn3m(Q²²⁴)结构. 实验发现, 鞘磷脂的含量对DEPE膜的结构有一定的影响, 随着鞘磷脂浓度的增加, 混合脂体系的液晶态结构发生了由Im3m(Q²²⁹)到Pn3m(Q²²⁴)的变化. 神经节苷脂(Gm1)的含量对混合脂体系的液晶态结构也有一定的影响, 当神经节苷脂(Gm1)含量达到某一临界值时, 混合脂体系的液晶态结构发生了从Im3m(Q²²⁹)到Pn3m(Q²²⁴) 的变化. 当DEPE-Shp-Gm1超分子聚集体中含有胆固醇时, 胆固醇的极性头部(—OH)与磷脂酰乙醇胺(DEPE)、鞘磷脂(Shp)、神经节苷脂(Gm1)的极性头部通过氢键相互作用形成液晶态立方相Im3m(Q²²⁹)结构, 再通过疏水/亲水相互作用形成稳定的Pn3m (Q²²⁴)结构.     

Free and bound water influence on Spirulina platensis survival

The heat effects arising at heating of Spirulina platensis cell culture containing different quantity of water (from 98.2 to 10.5 mass% H₂O) have been studied. The hydration of Sp. pl. cells determined by the method of microcalorimetry at 25°C (Δn) was equal to 0.32±0.02 g H₂O/g of dry biomass. The heat (–Q) evolved by cells in the temperature range 5–52°C decreased exponentially at decrease of mass% H₂O and reached zero value at 30.5±3.0 mass% H₂O. The total heat of cell denaturation did not change in the range 98.2–40.5 mass% H₂O and it sharply dropped at lower values of water.     

Study of Thermokinetic Properties of Sodium Selenite on Bacillus thuringiensis Cry B by Microcalorimetry

By using an LKB2277 Bioactivity Monitor, the power‐time curves of Bacillus thuringiensis Cry B at 28°C effected by Na₂SeO₃ were determined. Some parameters, such as growth rate constants k, inhibitory ratio I, the maximum beat production rate P_max, heat output Q, were obtained. Considering both the growth rate constant k and heat output Q, we found that a low concentration of Na₂SeO₃ had a promoting action on the growth of Bacillus thuringiensis Cry B, but a high concentration of Na₂SeO₃ had an inhibitory action. The toxicity of a toxicant can also be expressed as half inhibitory concentration IC₅₀ of toxicant, i. e., 50% effective in this inhibition. The value of IC₅₀ of Na₂SeO₃on Bacillus thuringiensis Cry B is 117 μg/mL. This microcalorimetric bioassay for cellular toxicity is based on metabolic heat evolution from cultured cells. The assay is quantitative, inexpensive, and versatile.     

Nitrogen Limitation in Neochloris oleoabundans: A Reassessment of Its Effect on Cell Growth and Biochemical Composition

The aim of this work was to reassess the effect of nitrogen limitation (from 0 to 1 mM nitrate), on the growth and the biochemical composition of Neochloris oleoabundans cultures, where only the CO₂ available in the air was provided. Slight differences in the initial nitrate concentration, even minimal increments of 0.2 mM, significantly modify the microalgal response towards nitrogen limitation. This stress condition reduced cell proliferation, but increased cell mass values due to the simultaneous accumulation of two storage compounds: lipids, which contained up to a 55.9 % of total fatty acids; and carbohydrates, which may be primarily composed by starch. The highest biomass and lipid productivities of 98.24 and 43.24 mg/l/day, respectively, were attained at an initial nitrate concentration of 0.6 mM. The theoretical annual projection, based on these productivities, allowed the estimation of the liquid fuel energy yields, which are comparable or even higher than those calculated for several biomass feedstocks such as corn, oil palm, sugarcane, or even fast growing grasses, confirming the potential of nitrogen-limited N. oleoabundans biomass as an appropriate feedstock for biofuel purposes.     

Biocalorimetry as a process analytical technology process analyser; robust in-line monitoring and control of aerobic fed-batch cultures of crabtree-negative yeast cells

Control of bioprocesses requires reliable and robust on- or in-line monitoring tools providing real-time information on process dynamics. Heat generation related to metabolic activity of living systems is currently gaining importance in bioprocess industry due to its non-invasive and essentially instantaneous characteristics. This study deals with monitoring and control of pure aerobic fed-batch cultures of three Crabtree-negative yeast strains, Kluyveromyces marxianus, Candida utilis and Pichia pastoris, based on in-line measured, metabolic heat flow signals. A high resolution biocalorimeter (BioRC1) was developed from a standard bench-scale heat flow calorimeter (RC1). The BioRC1 was equipped with in-line (dielectric spectroscopy, pH probe and dissolved oxygen probe) and at-line (exit gas analyser) sensors to characterise the growth behaviour of the yeast cells. Both metabolic heat flow and biomass profiles exhibited similar behaviour proving the significance of employing heat flow signal as a key-parameter for the system under investigation. A simple estimator for biomass concentration and specific growth rate was formulated based on heat flow values. In order to evaluate the potential of calorimetry as a reliable and powerful process monitoring tool, the robustness, reliability as well as the broad applicability of the developed estimators was assessed through comparison with off-line measurement techniques and showed promising results for general applicability with a wide range of bioprocesses.     

Microcalorimetric Studies on Influence of Sm^3＋, Dy^3＋ on Growth and Sporulation of Bacillus thuringiensis

By using an LKB-2277 Bioactivity Monitor and cycle-flow method, the thermogenic curves of aerobic growth for Bacillus thuringiensis cry Ⅱ strain at 28 ℃ have been obtained. The metabolic thermogenic curves of Bt cry Ⅱ contain two distinct parts: the first part reflects the changes of bacterial growth phase and the second part corresponds to sporulation phase. From these thermogenic curves in the absence or presence of Sm^3 , Dy^3 ions, the thermokinetic parameters such as the growth rate constants k. the interval time τ1, the maximum power PMAX1 and heat-output QLOG for log phase, the maximum power PMAX2 and heat-output QSTAT for stationary phase, the heat-output QSPOR for sporulation phase and total heat effects QT were calculated. Sm^3 and Dy^3 ions have promoting action on the growth of Bt cryⅡ in their lower concentration range, on the other hand, they have inhibitory action on the sporulation of Bt in their higher concentration range, It has also been found that the effects of Sm^3 and Dy^3 ions on Bt during the sporulation phase were far greater than those during the bacterial growth phase. It was concluded that the application of Bt for controlling insecticide could not be affected by the presence of the rare-earth elements in the environmental ecosystem.     

Characterization of chromium-induced apoptosis in cultured mammalian cells:: A differential scanning calorimetry study

The present study examined the cytotoxic effect of increasing Cr(VI) concentrations on cultured cells by a combination of biochemical methods and DSC, a novel use of DSC in the study of cell death. The characteristics of apoptotic cells are compared with normal cells. Chromatin in human epithelial-like L-41 cells has two thermal transitions at 100 and 105 °C. The heat from these endotherms is 90.5 ± 11.0 J/g DNA. The total heat of denaturation (Q_d) is 27.5 ± 3.5 J/g dry biomass. The heat evolved (−Q) is 15.6 ± 3.0 J/g dry biomass. The treatment of cells with 20 μM Cr(VI) for 2 and 4 h has not revealed any changes in heat of denaturation and heat evolution (−Q). However increased treatment time with Cr(VI) at 20 μM resulted in significant changes to the thermal profile and a sharp linear decrease of (−Q) and Q_d values. The Q_d and (−Q) values of cells treated with 20 μM Cr(VI) for 48 h are equal to 15.5 ± 2.0 and 2.1 ± 0.4 J/g dry biomass, respectively. The changes in chromatin conformation, Bax expression and the collapse of the mitochondrial membrane permeability coincide with the time point from which the action of chromium is irreversible.     

A microcalorimetric study of the biologic effect of Mn(II) on Bacillus thuringiensis growth

A microcalorimetric technique based on the bacterial heat-output was explored to evaluate the effect of Mn(II) on Bacillus thuringiensis. The power-time curves of the growth metabolism of B. thuringiensis and the effect of Mn(II) on it were studied using an LKB-2277 BioActivity Monitor, ampoules method, at 28°C. For evaluation of the results, the maximum peak-heat output power (P _max) in the growth phase, the growth rate constants (k), the log phase heat effects (Q _log ), and the total heat effect in 23 h (Q _T) for B. thuringiensis were determined. Manganese has been regarded as the essential biological trace element. Mn(II) of different concentration have different effects on B. thuringiensis growth metabolism. High concentration (800-1600 μg mL^-1) of Mn(II) can promote the growth of B. thuringiensis; low concentration (500-800 μg mL^-1) can inhabit its growth. This revised version was published online in August 2006 with corrections to the Cover Date.     

Axenic Cultures of Nitrosomonas europaea and Nitrobacter winogradskyi in Autotrophic Conditions: a New Protocol for Kinetic Studies

As a part of a natural biological N-cycle, nitrification is one of the steps included in the conception of artificial ecosystems designed for extraterrestrial life support systems (LSS) such as Micro-Ecological Life Support System Alternative (MELiSSA) project, which is the LSS project of the European Space Agency. Nitrification in aerobic environments is carried out by two groups of bacteria in a two-step process. The ammonia-oxidizing bacteria (Nitrosomonas europaea) realize the oxidation of ammonia to nitrite, and the nitrite-oxidizing bacteria (Nitrobacter winogradskyi), the oxidation of nitrite to nitrate. In both cases, the bacteria achieve these oxidations to obtain an energy and reductant source for their growth and maintenance. Furthermore, both groups also use CO₂ predominantly as their carbon source. They are typically found together in ecosystems, and consequently, nitrite accumulation is rare. Due to the necessity of modeling accurately conversion yields and transformation rates to achieve a complete modeling of MELiSSA, the present study focuses on the experimental determination of nitrogen to biomass conversion yields. Kinetic and mass balance studies for axenic cultures of Nitrosomonas europaea and Nitrobacter winogradskyi in autotrophic conditions are performed. The follow-up of these cultures is done using flow cytometry for assessing biomass concentrations and ionic chromatography for ammonium, nitrite, and nitrate concentrations. A linear correlation is observed between cell count and optical density (OD) measurement (within a 10?% accuracy) validating OD measurements for an on-line estimation of biomass quantity even at very low biomass concentrations. The conversion between cell count and biomass concentration has been determined: 7.1?×?10¹² cells g dry matter (DM)^?1 for Nitrobacter and 6.3?×?10¹² cells g DM^?1 for Nitrosomonas. Nitrogen substrates and products are assessed redundantly showing excellent agreement for mass balance purposes and conversion yields determination. Although the dominant phenomena are the oxidation of NH ₄ ⁺ into nitrite (0.95?mol?mol N^?1 for Nitrosomonas europaea within an accuracy of 3?%) and nitrite into nitrate (0.975?mol?mol N^?1 for Nitrobacter winogradskyi within an accuracy of 2?%), the Nitrosomonas europaea conversion yield is estimated to be 0.42?g DM mol?N^?1, and Nitrobacter winogradskyi conversion yield is estimated to be 0.27?g DM mol?N^?1. The growth rates of both strains appear to be dominated by the oxygen transfer into the experimental setups.