微波制样的应用

介绍了微波制样在分析化学中的应用，包括微波试样消解，微波加热水样测定CODcr，微波加热水解蛋白质测定各种氨基酸，以及微波萃取等。特别是对微波试样消解在应用中的一些问题，作了较详细的叙述。     

Alteration in photosystem II photochemistry of thylakoids isolated from senescing leaves of wheat seedlings

Wheat seedlings, grown for 7 days in the light, were allowed to senesce in the light or dark, and the change in the photosystem II (PS II) photochemistry of chloroplasts isolated from the primary leaves of these seedlings was investigated. The decrease in oxygen evolution and the fast fluorescence results indicated that the impairment of PS II in the leaves of seedlings senescing in the light was different from that in the leaves of seedlings senescing in the dark. Thermoluminescence studies showed a structural modification in the Q_B protein of chloroplasts isolated from leaves senescing in the light and an alteration in the S state transition of chloroplasts isolated from leaves senescing in the dark.     

Sol-gel Transition of Methylcellulose Solution in the Coexistence of Hexadecyltrimethylammonium Bromide and Sodium Chloride

The sol-gel transition of methylcellulose (MC) solution in the presence of NaCl and hexade-cyltrimethylammonium bromide (HTAB), together with MC/NaCl solution in the presence of HTAB and MC/HATB solution in the presence of NaCl, was investigated by the rheolog-ical measurements. It has been found that the sol-gel transition temperature of MC solution decreases linearly with the concentration of NaCl in solution but increases linearly with the concentration of HTAB in solution, respectively. However, the sol-gel transition temperature of MC/NaCl solution in the presence of HTAB keeps the same value, independent of theconcentration of HTAB in solution. On the other hand, the sol-gel transition temperature of MC/HTAB solution decreases linearly with the concentration of NaCl in solution. The experimental results suggest that, for MC/NaCl solution in the presence of HTAB, the salt-induced spherical micelles of HTAB should have formed in bulk solution. For MC solution in the absence of NaCl, no spherical micelles have been formed in bulk solution, though the concentration of HTAB in our experiment is almost one order of magnitude higher than the critical micelle concentration of HTAB in polymer-free solution. In fact, due to adsorption of HTAB on MC chains, the realconcentration of HTAB in bulk solution, is much less than the apparent concentration of HTAB dissolved in MC solution.     

Polymerization of N-carboxy–amino acid anhydrides in the solid state. II. Relation between polymerizability and molecular arrangement in L-leucine NCA and L-alanine NCA crystals

The polymerizability of N-carboxy–amino acid anhydrides (NCAs) of L -leucine and L -alanine was examined in the solid state and in solution. L -leucine NCA shows much higher reactivity in the solid state (when immersed in hexane) than in solution (in acetonitrile), but the opposite is true for L -alanine NCA. However, the two NCAs give similar values of apparent activation energy in each polymerization system. Rather high-molecular-weight polypeptides were obtained in the polymerization of L -leucine NCA in the solid state compared with those obtained in solution, while the molecular weight of polymers obtained from L -alanine NCA was higher in solution than in the solid state. IR spectra showed that α helices form mainly in the polymerization of both L -leucine NCA and L -alanine NCA in the solid state; a small amount of the β structure forms in the latter polymerization. X-ray diffraction and electron microscopy revealed that L -leucine NCA polymerizes predominantly along the c axis in the crystal, while the polymer chains grow in random directions in the crystal of L -alanine NCA. The difference can be explained by the molecular arrangement in the crystal. There are two requirements for high reactivity in the solid state: the five-membered rings of the monomer must form a layer structure and the polymer must occupy nearly the same space as the reacting monomer.     

Photoelectric effects on chlorophyll fluorescence of photosystem II in vivo. Kinetics in the absence and presence of valinomycin

Fluorescence induction curves (F(t)) in low intensity 1s light pulses have been measured in leaf discs in the presence and absence of valinomycin (VMC). Addition of VMC causes: (i) no effect on the initial fluorescence level Fo and the initial (O-J) phase of F(t) in the 0.01-1 ms time range. (ii) An approximately 10% decrease in the maximal fluorescence Fm in the light reached at the P level in the O-J-I-P induction curve. (iii) Nearly twofold increase in the rate and extent of the F(t) rise in the J-I phase in the 1-50 ms time range. (iv) A 60-70% decrease in the rise (I-P phase) in the 50-1000 ms time range with no appreciable effect, if at all, on the rate. System analysis of F(t) in terms of rate constants of electron transfer at donor and acceptor sides have been done using the Three State Trapping Model (TSTM). This reveals that VMC causes: (i) no, or very little effect on rate constants of e-transfer reactions powered by PSII. (ii) A manifold lower rate constant of radical pair recombination (k(-1)) in the light as compared to that in the control. The low rate constant of radical pair recombination in the reaction center (RC) in the presence of VMC is reflected by a substantial increase in the nonzero trapping efficiency in RCs in which the primary quinone acceptor (Q(A)) is reduced (semi-open centers). This causes an increase in their rate of closure and in the overall trapping efficiency. Data suggest evidence that membrane chaotropic agents like VMC abolish the stimulation of the rate constant of radical pair recombination by light. This light stimulation that becomes apparent as an increase in Fo has been documented before [Biophys. J. 79 (2000) 26]. It has been ascribed to effects of (changes in) local electric fields in the vicinity of the RC. The decrease of the I-P phase is attributed to a decrease in the photoelectric trans-thylakoid potential in the presence of VMC. Such effects have been hypothesized and illustrated.     

Grand canonical ensemble molecular dynamics simulation of water solubility in polyamide-6,6

Grand canonical ensemble molecular dynamics simulation is employed to calculate the solubility of water in polyamide-6,6. It is shown that performing two separate simulations, one in the polymeric phase and one in the gaseous phase, is sufficient to find the phase coexistence point. In this method, the chemical potential of water in the polymer phase is expanded as a first-order Taylor series in terms of pressure. Knowing the chemical potential of water in the polymer phase in terms of pressure, another simulation for water in the gaseous phase, in the grand canonical ensemble, is done in which the target chemical potential is set in terms of pressure in the gas phase. The phase coexistence point can easily be calculated from the results of these two independent simulations. Our calculated sorption isotherms and solubility coefficients of water in polyamide-6,6, over a wide range of temperatures and pressures, agree with experimental data.     

The heat of dissolution of uranyl nitrate in aqueous nitrate solutions

Conclusions A model has been suggested to explain the observed relationship between the measured heats of dissolution of uranyl nitrate in aqueous nitrate solutions and the concentration of the salting-out agent. The model describes the change in the structure of water in the solution with change in its concentration. On the one hand, a destruction of the water structure by ions occurs, which is weakened with increase in the distance from the ion, and leads to such irregularity in the distribution of water molecules in the solution that the mean number of molecules of water in unit volume is increased with increase in the distance from the ions. In experiments on the heat of dissolution this increase leads to increased hydration of the uranyl cation and reduction in the endothermicity of the dissolution with increase in the concentration of the solution. On the other hand, an interaction occurs between the ions of the salting-out agent and the water molecules in the solution, leading to the opposite result: There is an increase in the mean number of water molecules of the solution in unit volume in the direction of these ions. In experiments on the heat of dissolution this is revealed in the dehydration of the uranyl cation, and correspondingly in an increase in the endothermicity of the dissolution with increase in the concentration of the solution. The proposed model is in harmony with data on vapor pressure above the solutions (the relationship between the activity coefficient of the water and the concentration of the solution).Translated from Zhurnal Strukturnoi Khimii. Vol. 3, No. 2, pp. 143–150, March–April. 1962     

Direct determination of horseradish peroxidase encapsulated in liposomes by using luminol chemiluminescence.

Horseradish peroxidase (HRP) encapsulated in liposomes was directly detected by using luminol chemiluminescence (CL) with H2O2 without lysis of liposomes. At a low concentration of H2O2, the initial rate of HRP-catalyzed luminol CL in liposomes was slower than that of HRP-catalyzed luminol CL in a lipid-free bulk solution. The decrease in the initial rate of the CL reaction in liposomes was due to the membrane permeation of luminol and H2O2. At a high concentration of H2O2, the initial rate of the CL reaction in liposomes was the same as that in a lipid-free bulk solution. The CL measurement conditions in both a lipid-free bulk solution and in liposomes were optimized in the concentrations of luminol and H2O2 by measuring the CL response curves, in which only one peak appeared and the CL intensity was maximal. The CL intensity observed in HRP-catalyzed luminol CL in liposomes was a factor of seven greater than that observed in a lipid-free bulk solution. The CL intensity was dependent on the amount of HRP-encapsulated liposomes used. The detection limit in the direct detection of HRP encapsulated in liposomes was sensitive by a factor of 3 compared with that in HRP-catalyzed luminol CL in a lipid-free bulk solution.     

Determination of Penicillins in Milk by a Dual-Optrode Biosensor

Penicillins are the most frequently found antibiotic residues in milk, as they are commonly used for the treatment of bacterial infections in cows. In the present study, we introduce a method for the rapid detection of penicillin residues in raw milk based on the determination of glucose concentration in milk with a dual flow-through biosensor. The molar concentration of glucose in milk is typically over 500 times lower than the concentration of lactose and is highly dependent on the rate of lactose hydrolysis, which is catalyzed by β-galactosidase. Glucose concentrations in milk change with variation in the β-galactosidase activity. β-Galactosidase is an enzyme produced in the microbiota in milk and its activity is inhibited by benzylpenicillin. Spiking milk with benzylpenicillin lowers glucose concentrations in comparison to high-quality milk after short storage intervals. The presence of penicillin in the milk of treated animals resulted in decreased glucose concentrations in comparison with high-quality milk that contained no antibiotics. The glucose concentration in milk samples was followed by the system enabling the elimination of the effects of bacterial respiration in the output with reliable results in less than 1?min.     

Chemical reaction in binary mixtures near the critical region: thermal decomposition of 2,2'-azobis(isobutyronitrile) in CO2/ethanol

The effects of pressure and of the composition of the CO2/ethanol mixed solvent in the critical region on the kinetics of the decomposition of 2,2'-azobis(isobutyronitrile) (AIBN) were studied at 333.15 K. The rate constants (kd) in the mixed solvent far from the critical point and in liquid n-hexane and ethanol were also determined for comparison. It was found that kd is very sensitive to pressure in the mixed solvent near the critical point. However, in the mixed solvent outside the critical region kd is nearly independent of pressure. Interestingly, kd in the mixed solvent in the critical region can be higher than that in ethanol at the same temperature, suggesting that no significant enhancement in the reaction rate by a small pressure change in the critical region of the mixed solvent can be achieved by changing the composition of the liquid solvent in the traditional way. Transition-state theory can predict kd in the mixed solvent far from the critical point and in the liquid solvents well. However, it cannot predict kd in the mixed solvent in the critical region. The special intermolecular interaction between the solvent and the reaction species may contribute to this interesting phenomenon. This work also shows that if pure CO2 or ethanol are used as solvents, the reaction cannot be carried out in the critical region of the solvents at the desired temperature, while it can be conducted in the critical region of the mixed solvent of suitable composition, where the solvent is highly compressible.     

Use of <Superscript>82</Superscript>Br and <Superscript>131</Superscript>I radionuclides in studies of goitrogenic effects of exogenous bromide

The interference of excessive bromide intake with iodine metabolism in the rat was studied using ⁸²Br and ¹³¹I radionuclides. Mainly lactating rat dams and their pups were used, in addition to adult male rats, in the present studies. Particularly, the influence of high bromide intake in lactating rat dams on the transfer of iodine and bromide to suckling young through breast milk was evaluated. The induction of hypothyroid status in the pups by high bromide intake in the mothers was proved unambiguously. Excessive bromide in lactating rat dams caused a marked decrease in plasma levels of thyroid hormones both, in the mothers and in their sucklings. The effects of an enhanced bromide intake on the thyroid function in relation to iodine status in the animals were also followed. Marked goitrogenic and thyrotoxic effects of excessive bromide in adult rats were significantly enhanced under the conditions of simultaneous iodine deficiency in the experimental animals.     

Anomalous low-frequency electro-optic behavior of ferric oxide particles in the presence of poly(ethylene oxide)

Electro-optic techniques were used to investigate the influence of poly(ethylene oxide) (PEO) on the surface electric state of positively charged oxide particles. The variations in particle electrophoretic mobility of beta-FeOOH particles in the presence of PEO indicate significant changes in the surface electric state of the particles in the concentration interval of PEO 10(-2)-10(-1) g dm(-3). The electro-optic results for the same conditions were unexpected: no significant difference is observed in the value and the relaxation frequency of particle electric polarizability in the frequency domain of the alpha-relaxation (detected in the kilohertz range); particle rotational relaxation time also remains unchanged; considerable changes are detected only in the relaxation interval of particle rotation (detected in the hertz range). The obtained results reject the possibility of the formation on the particle surface of a thick polymer layer. A thin adsorption layer cannot explain the significant decrease in particle electrophoretic mobility. The variations in electrophoretic mobility are well correlated with the effects in the domain of particle rotation. A possible explanation of the observed effects is proposed, based on our previous investigations of the effects in the low-frequency domain. The presented results demonstrate that the important information on the electrokinetic charge distribution is not found in the domain of the alpha-dispersion, but in the domain of particle rotation.     

Independence of the lethal actions of glucocorticoids on lymphoid cells from possible hormone effects on calcium uptake

We have examined the possibility that hormone-induced increases in calcium uptake might initiate the lethal actions of glucocorticoids in two types of lymphoid cells. Hormone-induced increases in nuclear fragility are used as the measure of hormone action, since in both rat thymus cells and in mouse P1798 lymphosarcoma cells increased nuclear fragility (the inability of nuclei to survive lysis of the cells by hypotonic shock) precedes other indices of cellular deterioration by several hours. In the case of the tumor cells, those from corticosteroid-sensitive lines are less able to withstand incubation in vitro than resistant cells. Such differences in cell survival are predicted both by earlier changes in nuclear fragility and also by differences in calcium uptake. However, there is no detectable early glucocorticoid effect on calcium uptake that precedes or coincides with the substantial hormone-induced increases in nuclear fragility that develop in the sensitive cells by 2 h. In rat thymus cells the absence of calcium in the medium does prevent some of the increase in nuclear fragility and cell disintegration that occurs spontaneously during incubation in vitro. Nevertheless, when cells are exposed to hormones the glucocorticoid effect on nuclear fragility develops in the absence of calcium and is similar in magnitude to that seen in the presence of calcium. We conclude that calcium seems to enhance the spontaneous deterioration of lymphoid cells, and there is a large increase in calcium uptake that occurs as cells deteriorate. It nevertheless seems unlikely that hormone-induced changes in calcium uptake initiate the lethal actions of glucocorticoids. The data also support a proposal made earlier [2] that resistance to glucocorticoids in tumor cells may develop by the selection of cells with hardier membranes.     

Enthalpy of solution of 1,4-naphthoquinone in CO2 + n-pentane in the critical region of the binary mixture: mechanism of solubility enhancement

The enthalpy of solution (Delta(solv)H(m)) and solubility of 1,4-naphthoquinone in CO(2) + n-pentane were measured at 308.15 K in the critical region of the binary fluid. In order to study the effect of phase behavior of the mixed solvent on Delta(solv)H(m), the experiments were carried out in the supercritical (SC) and subcritical region of the binary solvent. The density of the mixed solvent in different conditions was determined. The isothermal compressibility (K(T)) of the mixed solvent, and the partial molar volume (V(n-pentane)) of n-pentane in the solution were calculated. It was demonstrated that the Delta(solv)H(m) was negative in all conditions. Delta(solv)H(m) is nearly independent of pressure or density in all the solvents in a high-density region, in which compressibility of the solvent is very small; this indicates that the intermolecular interaction between the solvent and the solute is similar to that for liquid solutions. It is very interesting that Delta(solv)H(m) in the mixed SC fluid differs from the Delta(solv)H(m) in mixed subcritical fluids. The absolute value of Delta(solv)H(m) in the mixed SC fluid is close to that in pure SC CO(2) in the high-density region, and is much lower than that in pure SC CO(2) in the low-density region. In the mixed subcritical fluids, the Delta(solv)H(m) is also close to that in the pure CO(2) in the high-density region. However, at the same density, the absolute value of Delta(solv)H(m) in the binary subcritical fluid is larger than that in pure CO(2) in the high-compressible region of the mixed solvent. The main reason for this is that the degree of clustering in the SC solutions is small at the density in which the degree of clustering is large in the subcritical solutions. It can be concluded that solubility enhancement by n-pentane in the mixed SC fluid is entropy driven. In contrast, the solubility enhancement by n-pentane in subcritical fluids is enthalpy driven. The intermolecular interaction in the SC solutions and subcritical solutions can be significantly different even if their densities are the same.     

Riboflavin and UV-Light Based Pathogen Reduction: Extent and Consequence of DNA Damage at the Molecular Level

We are developing a technology based on the combined application of riboflavin (RB) and light for inactivating pathogens in blood products while retaining the biological functions of the treated cells and proteins. Virus and bacteria reduction measured by tissue culture infectivity or colony formation with UV light alone and in combination with RB yield equivalent results. The effects of RB as a sensitizing agent on DNA in white cells, bacteria and viruses in combination with UV light exposure have been evaluated. UV-mediated DNA degradation in Jurkat T cells and leukocytes in plasma as measured by the FlowTACS assay was significantly increased in the presence of RB. Agarose gel electrophoretic analysis of DNA in Escherichia coli and leukocytes in plasma demonstrated enhanced DNA degradation in the presence of RB. UV light in combination with RB prevented the reactivation of lambda phage compared with samples irradiated in the absence of RB. UV-mediated oxidative damage in calf thymus DNA was also enhanced in the presence of RB. These observations clearly demonstrate that the presence of RB and UV light selectively enhances damage to the guanine bases in DNA. These data also suggest that the type and extent of damage to DNA for virus in the presence of RB and light make it less likely to be repaired by normal repair pathways available in host cells.     

MODIFYING FACTORS OF THE CELLULAR CONCENTRATION OF PHOTOLYASE MOLECULES IN Saccharomyces cerevisiae CELLS–I. EFFECTS OF TEMPERATURE AND LIGHT

Abstract— The effects of temperature and light on the cellular concentration of photoreactivating enzyme (PRE) molecules in haploid Saccharomyces cerevisiae cells were investigated. (1) Temperature effect: The number of active PRE molecules per cell (N_PRE) in cells grown at 37°C was about 13% of that grown at 23°C, although the amount of proteins per cell remained the same. (2) Light effect: N_PRE in cells grown in light was about 2.8 times larger than that grown in the dark. The value of N_PRE in cells grown in the light decreased more rapidly during holding in buffer in the dark than in the light. The N_PRE decrease during holding in buffer in the dark was more rapid in cells grown in the light than grown in the dark. A comparable decrease was observed after holding in buffer in the presence of cycloheximide. (3) In cells harboring a plasmid containing the gene PHR1, N_PRE was larger in cells grown at 23 than at 30°C.     

Localisation and accumulation of a new carotenoporphyrin in two primary tumour models

We have investigated the tumour-localising properties and in vivo fluorescence kinetics of a hexamethoxylated carotenqporphyrin (CP6) in two primary tumour models: UV-B-induced early skin cancer in hairless mice and chemically induced mucosal dysplasia in the rat palate. CP6 fluorescence kinetics are investigated by measuring in vivo fluorescence spectra and images of the mouse skin and the rat palate at different time points after injection. For the tumour-localising properties, microscopic phase-contrast and fluorescence images are recorded. The in vivo fluorescence kinetics in the mouse skin show localization of CP6 in the tumours. However, fluorescence microscopy images show that CP6 localises in the dermis and structures that are not related to the malignant transformation of the mouse skin. The fluorescence kinetics in the rat palate show a significant correlation between the degree of malignancy and the CP6 fluorescence build-up time in the palate. The microscopic images show that CP6 fluorescence localises in the connective tissue and not in the dysplastic epithelium. In conclusion, CP6 does not localise preferentially in (pre-) cancerous tissue in the two primary tumour models studied here, in contrast to reports about localisation of carotenoporphyrins in transplanted tumours. However, the CP6 build-up time in rat palates correlates with the degree of malignancy and this might possibly be a useful parameter in tumour detection.     

基于MALDI质谱成像技术分析莲子中代谢物的组织分布

该文建立了一种可对莲子中多种代谢物进行高覆盖分析的基质辅助激光解吸附质谱成像(MALDI-MSI)方法,实现了莲子中生物碱类、黄酮类、氨基酸类、脂肪酸类、有机酸类、胆碱类、磷脂类等多种代谢物的组织原位可视化表征。结果表明,生物碱类代谢物主要分布在莲子胚芽中;黄酮类代谢物主要分布在莲子胚芽和种皮中;氨基酸类代谢物在莲子子叶中的含量显著高于莲子胚中;脂肪酸类代谢物在莲子不同组织中的分布差异很小;胆碱类代谢物在莲子胚芽和莲子子叶底部的含量更高,甘油磷酸胆碱在莲子子叶顶部的含量更高;有机酸类代谢物以及绝大多数磷脂类化合物在莲子子叶中的含量高于莲子胚。该研究为评价莲子药物质量、探究莲子中化合物的时-空代谢网络提供了新的技术支持。     

Electric-field-induced changes in absorption and emission spectra of CdS nanoparticles doped in a polymer film

Electric-field-induced changes in absorption and emission spectra of colloidal CdS nanoparticles ranging in size from 1.0 to 5.0 nm in diameter have been measured by using electric field modulation spectroscopy. The analysis of the electroabsorption spectra indicates that the dipole moment in the first exciton state becomes larger with increasing particle size. The presence of the large dipole moment following photoexcitation into the first exciton band suggests that the CdS nanoparticles have large CT character in the first exciton state. The quantum yields both of the exciton emission and of the trap emission are markedly reduced by application of an electric field. On the basis of the direct measurements of the field-induced change in emission decay profile, it is suggested that the field-induced de-enhancement of these emission yields results from the field-induced decreases both in lifetime and in initial population of each emitting state. It is also found that the emission intensity of CdS nanoparticles increases under the UV light irradiation in air and decreases in a vacuum condition and that fluorescence lifetime in the former case is longer than that in the latter. This enhancement and de-enhancement process in emission intensity is almost reversible at least in several cycles.     

Changes in Porosity and Amounts of Adsorbed Water in Sol-Gel Derived Porous Silica Films with Heat Treatment

Porous silica films were obtained by a heat treatment of poly(ethyleneglycol)-containing gels for micro-patterning. Changes in porosity and the amount of adsorbed water with the heat treatment temperature in the silica films were evaluated from the refractive index of the films. The apparent refractive index of the silica films in an ambient atmosphere was much higher than the intrinsic refractive index of the film due to the large amount of adsorbed water in the films. The amount of adsorbed water in the films decreased with an increase in the heat treatment temperature and became almost zero after a heat treatment at 850°C. The decrease in the amount of adsorbed water in the films was caused by the decrease in silanol groups, which acted as the sites for water adsorption in the films. The drastic decreases in thickness and in porosity of the films at temperatures from 850 to 950°C occurred after the disappearance of silanol groups in the films. The densification of the films due to collapse of the pores at high temperatures should have resulted from the viscous flow of silica network.