Photocatalytic antibacterial effects on TiO2-anatase upon UV-A and UV-A/VIS threshold irradiation

Photocatalysis mediated by the anatase modification of titanium dioxide (TiO₂) has shown antibacterial effects in medical applications. The aim of this study was to investigate the possibility of expanding the excitation wavelengths for photocatalytic antibacterial effects from ultraviolet (UV) into the visible light range. After deposition of salivary pellicle and adhesion of Streptococcus gordonii on anatase, different irradiation protocols were applied to induce photocatalysis: ultraviolet A (UV-A) > 320 nm; ultraviolet/visible (UV-A/VIS) light > 380 nm and > 390 nm; and VIS light 400–410 nm. A quartz crystal microbalance with dissipation (QCM-D) tests and microscopic examination were used to observe the photoinduced antibacterial effects. Salivary pellicle could be photocatalytically decomposed under all irradiation protocols. In contrast, effective photocatalytic attack of bacteria could be observed by UV-A as well as by UV-A/VIS at 380 nm < λ < 390 nm only. Wavelengths above 380 nm show promise for in situ therapeutic antifouling applications.     

Conformational changes of beta-lactoglobulin induced by anionic phospholipid

Conformational changes of beta-lactoglobulin (beta-LG) induced by anionic phospholipid (dimyristoylphosphatidylglycerol, DMPG) at physiological conditions (pH 7.0) have been investigated by UV-VIS, circular dichroism (CD) and fluorescence spectra. The experimental results suggest that beta-LG-DMPG interactions cause beta-LG a structural reorganization of the secondary structure elements accompanied by an increase in alpha-helical content, and a loosening of the protein tertiary structure. The interaction forces between beta-LG and DMPG are further evaluated by fluorescence spectra. The fluorescence spectral data show that conformational changes in the protein are driven by electrostatic interaction at first, then by hydrophobic interaction between a protein with a negative net charge and a negatively charged phospholipid.     

Spectroscopic properties of dissolved humic substances - a reflection of land use history in a fen area

The elemental composition and spectroscopic properties of dissolved fulvic acids isolated from different sampling media (topsoil, ground and surface water) of a natural fen area (high portion of organic soils) were examined to reveal the effects of land use history. These effects need to be known if dissolved humic substances are to be a major factor in identifying the impact of present and future changes in land use. Dissolved fulvic acids (topsoil, groundwater) from highly degraded peatlands (due to a long-term agricultural use) exhibit lower C/N ratios, higher absorption in the UV spectra, and higher absorption at 1,620 cm^–1 in the FTIR spectra compared with fulvic acids from relatively intact peatlands. These properties illustrate that long-term agricultural use with high inputs results in increased aromatic structures and a further humification of dissolved fulvic acids due to very strong peat decomposition compared with relatively intact peatlands. Synchronous fluorescence spectra also indicate the higher level of aromatic structures within fulvic acids isolated from sites with long-term agricultural use (high peat decomposition) compared with a land use history resulting in a lower peat decomposition. The different sources of fulvic acids in surface water (precipitation, runoff, interflow, groundwater) are the main reason for these effects not being detected in fulvic acids isolated from surface water. Short-term changes in land use characterized by a transition from crop farming to an unimproved grassland were found not to affect the spectroscopic properties of dissolved fulvic acids. A humification index deduced from the synchronous fluorescence spectra is proposed. We have strong evidence that dissolved humic substances indicate changes in the environmental conditions (both anthropogenic and natural) of wetlands with a high proportion of organic soils.     

1H- and 13C-NMR, FTIR, UV-VIS, ESI-MS, and PM5 studies as well as emission properties of a new Schiff base of gossypol with 5-methoxytryptamine and a new hydrazone of gossypol with dansylhydrazine

A new Schiff base of gossypol with 5-methoxytryptamine (GSTR) and a new hydrazone of gossypol with dansylhydrazine (GHDH) have been synthesized and studied by Fourier transform infrared (FTIR), 1H and 13C nuclear magnetic resonance (NMR), ultraviolet-visible (UV-VIS), electrospray ionization-mass spectroscopy (ESI-MS) as well as the parametric method PM5. The spectroscopic methods have provided clear evidence that GSTR exists in chloroform solution as an enamine-enamine tautomer, whereas GHDH is present in chloroform as a N-imine-N-imine tautomer. The fluorescence spectra of both compounds indicate that their quantum yield of fluorescence is increased by one or two orders of magnitude compared to that of pure gossypol. The ESI-MS spectra of the 1:1 mixtures of GSTR or GHDH with formic acid have demonstrated that both compounds exist as protonated monomers in the gas phase, whereas GHDH can also exist in a stable protonated dimeric structure. The structures of the stable tautomers are calculated and visualized using the PM5 semiempirical method. The intra- and intermolecular hydrogen bonds within these structures are discussed.     

VIS2FIX: a high-speed fixation method for immuno-electron microscopy

Immuno-transmission electron microscopy (TEM) is the technique of choice for high-resolution localization of proteins in fixed specimen. Here we introduce 2 novel methods for the fixation of sections from cryo-immobilized samples that result in excellent ultrastructural preservation. These high-speed fixation techniques, both called VIS2FIX, allow for a reduction in sample preparation time from at least 1 week to only 8 h. The methods were validated in immuno-TEM experiments on THP-1 monocytes, human umbilical vein endothelial cells (HUVECs) and Madin-Darby canine kidney (MDCK-II) cells. The fixation and retention of neutral lipids is demonstrated, offering unique prospects for the application of immuno-TEM in the lipidomics field. Furthermore, the VIS2FIX methods were successfully employed in correlative fluorescence and electron microscopy.     

Reversible control of F1-ATPase rotational motion using a photochromic ATP analog at the single molecule level

Motor enzymes such as F₁-ATPase and kinesin utilize energy from ATP for their motion. Molecular motions of these enzymes are critical to their catalytic mechanisms and were analyzed thoroughly using a single molecule observation technique. As a tool to analyze and control the ATP-driven motor enzyme motion, we recently synthesized a photoresponsive ATP analog with a p-tert-butylazobenzene tethered to the 2′ position of the ribose ring. Using cis/trans isomerization of the azobenzene moiety, we achieved a successful reversible photochromic control over a kinesin-microtubule system in an in vitro motility assay. Here we succeeded to control the hydrolytic activity and rotation of the rotary motor enzyme, F₁-ATPase, using this photosensitive ATP analog. Subsequent single molecule observations indicated a unique pause occurring at the ATP binding angle position in the presence of cis form of the analog.     

Characterization of the primary photochemistry of proteorhodopsin with femtosecond spectroscopy

Proteorhodopsin is an ion-translocating member of the microbial rhodopsin family. Light absorption by its retinal chromophore initiates a photocycle, driven by trans/cis isomerization, leading to transmembrane translocation of a proton toward the extracellular side of the cytoplasmic membrane. Here we report a study on the photoisomerization dynamics of the retinal chromophore of proteorhodopsin, using femtosecond time-resolved spectroscopy, by probing in the visible- and in the midinfrared spectral regions. Experiments were performed both at pH 9.5 (a physiologically relevant pH value in which the primary proton acceptor of the protonated Schiff base, Asp⁹⁷, is deprotonated) and at pH 6.5 (with Asp⁹⁷ protonated). Simultaneous analysis of the data sets recorded in the two spectral regions and at both pH values reveals a multiexponential excited state decay, with time constants of ∼0.2 ps, ∼2 ps, and ∼20 ps. From the difference spectra associated with these dynamics, we conclude that there are two chromophore-isomerizaton pathways that lead to the K-state: one with an effective rate of ∼(2 ps)⁻¹ and the other with a rate of ∼(20 ps)⁻¹. At high pH, both pathways are equally effective, with an estimated quantum yield for K-formation of ∼0.7. At pH 6.5, the slower pathway is less productive, which results in an isomerization quantum yield of 0.5. We further observe an ultrafast response of residue Asp²²⁷, which forms part of the counterion complex, corresponding to a strengthening of its hydrogen bond with the Schiff base on K-state formation; and a feature that develops on the 0.2 ps and 2 ps timescale and probably reflects a response of an amide II band in reaction to the isomerization process.     

野生火棘果红色素提取工艺优化

目的:通过正交实验,确定火棘果红色素的最佳提取工艺。方法:以贵州野生火棘果为原料,首先,以柠檬酸的乙醇溶液为提取剂,在400～800nm范围内对提取液的吸收光谱进行扫描,确定其最大吸收波长;然后,以最大吸收波长处的吸光度为衡量指标,确定最佳提取剂;再次,探讨了提取次数、提取温度、提取时间、料液比等单因素对火棘果红色素提取效果的影响;最后,在单因素实验结果的基础上,通过L9(34)正交实验,确定红色素的最佳提取工艺。结果:1.在400～800nm范围内,火棘果红色素的最大吸收波长为530nm;2.最佳提取剂为9%柠檬酸乙醇溶液;3.火棘果红色素的最佳提取工艺为,料液比1∶10,提取时间60min,提取次数2次,提取温度40℃。     

Further characterization of two different,reversible aldehyde oxidoreductases from Clostridium formicoaceticum,one containing tungsten and the other molybdenum

The tungsten- and the molybdenum-containing aldehyde oxidoreductases from Clostridium formicoaceticum show, for aldehydes, K _m values<30 M and K _i values of millimolar concentrations. The tungsten-containing aldehyde oxidoreductase is inactivated to 50% by 3 mM KCN within 1 min, by 1 mM ferricyanide within 5 min, and by 0.05 mM chloralhydrate within 30 s. The molybdenum-containing AOR shows 50% inactivation within 1 min only with 70 mM KCN. The tungsten-containing enzyme is very sensitive to oxygen, especially in the reduced state, whereas the molybdenum-containing enzyme exhibits only moderate oxygen sensitivity without being markedly influenced by the redox state of the enzyme. The tungsten in the aldehyde oxidoreductase is bound to a pterin cofactor (Wco) of the mononucleotide form that is known for molybdopterin cofactor (Moco). The nature of the molybdenum cofactor in the molybdenum-containing aldehyde oxidoreductase is still unclear. The UV/VIS spectrum of the tungsten-containing aldehyde oxidoreductase shows a broad absorption in the range of 400 nm with a millimolar absorption coefficient of 18.1 (reduced form) and 24.8 (dehydrogenated form) at 396 nm. The epr spectrum exhibits two different W(V) signals with the following g values for signal A: 2.035, 1.959, 1.899 and signal B: 2.028, 2.017, 2.002. Dithionite-reduced enzyme shows signals of 4Fe–4S or 2Fe–2S clusters. Initial rate studies with different substrates for the carboxylate reduction led to a Bi Uni Uni Bi mechanism.Abbreviations AOR aldehyde oxidoreductase - NH ₂ CO-MV 1,1-carbamoylmethylviologen - MV methylviologen - TMV 1,1,2,2-tetramethylviologen