Determination of absolute configuration and binding efficacy of benzimidazole-based FabI inhibitors through the support of electronic circular dichroism and MM-GBSA techniques

We have previously reported benzimidazole-based compounds to be potent inhibitors of FabI for Francisella tularensis (FtFabI), making them promising antimicrobial hits. Optically active enantiomers exhibit markedly differing affinities toward FtFabI. The IC₅₀ of benzimidazole (?)-1 is ～100× lower than the (+)-enantiomer, with similar results for the 2 enantiomers. Determining the absolute configuration for these optical compounds and elucidating their binding modes is important for further design. Electronic circular dichroism (ECD) quantum calculations have become important in determining absolute configurations of optical compounds. We determined the absolute configuration of (?)/(+)-1 and (?)/(+)-2 by comparing experimental spectra and theoretical density functional theory (DFT) simulations of ECD spectra at the B3LYP/6-311+G(2d, p) level using Gaussian09. Comparison of experimental and calculated ECD spectra indicates that the S configuration corresponds to the (?)-rotation for both compounds 1 and 2, while the R configuration corresponds to the (+)-rotation. Further, molecular dynamics simulations and MM-GBSA binding energy calculations for these two pairs of enantiomers with FtFabI show much tighter binding MM-GBSA free energies for S-1 and S-2 than for their enantiomers, R-1 and R-2, consistent with the S configuration being the more active one, and with the ECD determination of the S configuration corresponding to (?) and the R configuration corresponding to (+). Thus, our computational studies allow us to assign (?) to (S)- and (+) to (R)- for compounds 1 and 2, and to further evaluate structural changes to improve efficacy.     

Identification of cis/trans isomers of bis(acetylacetonato)nickel(II) complexes in solution based on electronic spectra

Electronic spectra of Ni(acac)₂ were studied in acetone, DMF, and some other solvents for the purpose of identifying the cis/trans isomers from the spectra (acac⁻ = acetylacetonate anion). The spectral components were investigated in the spin-allowed transition bands, and a relationship was found between the spectral pattern and the cis/trans isomers. According to this relationship, it was concluded that the cis isomer was formed in DMF and in N-methylformadide, whereas the trans isomer was formed in acetone and in pyridine. Based on the DFT computation, the cis-[Ni(acac)₂(DMF)₂] was found to be stabilized by intramolecular hydrogen bonds between acetylacetonate and DMF.     

Substance Flow Analysis of Mercury Intentionally Used in Products in the United States

Mercury-containing products release mercury (Hg) throughout their lifecycles, frequently in ways that are difficult to measure directly. Therefore, there are considerable uncertainties about the magnitude of mercury releases associated with products, about which products and which release pathways contribute the most to mercury releases, and about the likely impact on mercury releases of various possible interventions in the mercury content of products or in the management of mercury-containing wastes. This article presents an effort to use substance flow analysis to develop improved estimates of the environmental releases caused by mercury-containing products and to provide policy-makers with a better understanding of opportunities for reducing releases of mercury caused by products.     

Mitotic activity in wheat shoot apical meristems: effect of dissection to expose the apical dome

An efficient method, less laborious than histological procedures, is described to screen relatively large numbers of shoot apices for mitotic activity. Mitotic activity of shoot apices of Triticum aestivum L. was observed by differential interference contrast (DIC) microscopy of apices infiltrated with a clearing fluid (chloral hydrate/phenol/lactic acid/dibutylphthalate/benzyl benzoate). Serial optical sections were viewed through entire vegetative apical domes and floral primordia. In vegetative shoots, mitotic cells were observed throughout the light and dark cycles of plants maintained in either 8 or 16 h photoperiods. Mitotic activity was lower in the dark phase and increased through the light cycle in both photoperiods. Cells in L1 and L2 layers at the summit of the apex were mitotically active and contributed to the developing shoot and floral structures. Thus, cells in L2 at the summit of vegetative apices are valid targets for transformation leading subsequently to modified germ line cells. Dissections to expose apices for DNA delivery inhibited mitotic activity; recovery periods greater than 48 h would be needed for restoration of normal activity. This suggests that a period of recovery from dissection would be beneficial for attempts at integrative transformation of apical cells.     

Significance of treatment interval and DNA repair in the enhancement of viral transformation by chemical carcinogens and mutagens

Treatment of Syrian hamster embryo cells with diverse classes of chemical carcinogens enhanced transformation by a carcinogenic simian adenovirus, SA7. Optimal enhancement was a function of time of chemical addition in relation to time of virus addition and cell transfer. Aflatoxin B₁ (AFB₁) and the polycyclic hydrocarbons, benzo(a)pyrene (B(a)P), 3-methylcholanthrene (MCA), and 7,12-dimethylbenz(a)anthracene (DMBA) enhanced SA7 transformation when added prior to virus, but inhibited transformation when added after virus adsorption and cell transfer. The enhancement of SA7 transformation was maximal when cytosine arabinoside, caffeine and 6-acetoxy-benzo(a)pyrene (6-ac-B(a)P) were added after virus, but minimal when added before virus. A third class of chemicals, including β-propiolactone (β-PL), methyl methanesulfonate (MMS), N-acetoxy-2-acetylaminofluorene (Ac-AAF), N-methyl-N′-nitro-N-nitrosoguanidine (MNNG), and methylazoxymethanol acetate (MAM-ac), enhanced SA7 transformation added before, or after, virus inoculation and cell transfer. All chemicals, which induced changes in DNA sedimentation in alkaline sucrose gradients and unscheduled DNA (repair) synthesis in hamster cells, increased the frequency of SA7 transformation. However, several chemicals such as dibenz(a,h)anthracene (DB(a,h)A), benzo(e)pyrene (B(e)P), cytosine arabinoside, and caffeine enhanced SA7 transformation but did not induce DNA sedimentation changes or repair. Chemicals that cause DNA damage, which can be repaired by hamster cells, may enhance viral transformation by providing additional sites for integration of viral DNA during the repair process. Chemicals that apparently do not induce DNA repair synthesis may enhance viral transformation by incorporation of viral DNA into gaps in cell DNA at sites of unrepaired damage during scheduled DNA synthesis.     

点格局分析中边缘校正的一种新算法及其应用

在点格局分析中,通常选取一个矩形作为研究区域,而K(d)函数估计值的方差倾向于随着距离尺度的增加而增加.作为一种粗略的指导,距离尺度的最大值一般为矩形最小边长度的一半.在这种情况下,边缘校正的权重最小值为0.25.通过在校正圆上等弧长取点,用校正圆上落在研究区域之内的点数除以整个校正圆上的点数,作为边缘校正权重的近似值.点数越多,这种近似算法越接近传统的精确算法.这种近似算法不仅适用于计算研究区域为矩形的边缘校正权重,而且适用于计算研究区域为任意多边形的边缘校正权重.此外,当矩形研究区域中点事件的信息不足时,这种算法可以允许计算接近到距离尺度的上限(即矩形对角线长度的一半)对应的K(d)函数.     

Comparison of soybean cultivars for enhancement of the polyamine contents in the fermented soybean natto using Bacillus subtilis (natto)

Polyamines have beneficial properties to prevent aging-associated diseases. Raw soybean has relatively high polyamine contents; and the fermented soybean natto is a good source of polyamines. However, detailed information of diversity of polyamine content in raw soybean is lacking. The objectives of this study were to evaluate differences of polyamines among raw soybeans and select the high polyamine-containing cultivar for natto production. Polyamine contents were measured chromatographically in 16 samples of soybean, which showed high variation among soybeans as follows: 93–861 nmol/g putrescine, 1055–2306 nmol/g spermidine, and 177–578 nmol/g spermine. We then confirmed the high correlations of polyamine contents between raw soybean and natto (r = 0.96, 0.95, and 0.94 for putrescine, spermidine, and spermine, respectively). Furthermore, comparison of the polyamine contents among 9 Japanese cultivars showed that ‘Nakasen-nari’ has the highest polyamine contents, suggesting its suitability for enhancement of polyamine contents of natto.     

Modification by metals, sulphydryl reagents and cyanide of the particle stimulated enhancement of oxygen consumption in bovine granulocytes

A simple, rapid method for the isolation of neutrophil-enriched leucocyte suspensions from bovine blood is described. The capacity of these cells to produce a particle stimulated increase in oxygen consumption deteriorated during a period of storage of the cells whilst the viability of the cells remained unchanged. Potassium cyanide inhibited the basal oxygen consumption but enhanced the stimulated respiratory burst. Zinc ions also enhanced this respiratory burst but ferric and manganous ions did not. The lipid-soluble non-haem iron chelator, 4,4,4-trifluoro-1-(2 thienyl)-1,3-butanedione, preferentially inhibited the particle stimulated type of oxygen consumption, as did the sulphydryl reagents, N-ethyl maleimide and diazine dicarboxylic acid bis-dimethyl amide. These data allow us to consider that zinc ions may play a role in the respiratory burst associated with phagocytosis and that iron-sulphur interactions may be important in oxygen consuming reactions.     

Conservation of the alternative oxidase and enhancement of cyanide-resistant respiration in suspension-cultured pear fruit cells by inhibitors of macromolecular synthesis

The contribution of the alternative pathway to the respiration of suspension-cultured pear ( Pyrus communis cv. Passa Crasanne) cells was enhanced, often severalfold, within 2 to 4 days following the addition of cycloheximide, actinomycin D, or 2-(4-methyl-2,6-dinitroanalino)- N -methyl propionamide (D-MDMP). Concomitant inhibition of cellular protein synthesis by cycloheximide and actinomycin D was transient and incomplete. However, inhibition by D-MDMP was virtually complete (>97%) and persisted over several days. [³⁵S]-labelling and polyacrylamide gel separation indicated that cycloheximide precluded the appearance of discernable new proteins in mitochondria. Probes with monoclonal antibodies revealed a conservation of alternative oxidase protein levels in the mitochondria of inhibitor-treated cells. The data, appraised within the complexities of cell-culture dynamics, lead to the conclusion that the observed increases in capacity for cyanide-resistant respiration are the consequence, likely indirect, of inhibited protein synthesis with resultant retention and activation of constitutive alternative oxidase.     

Emission enhancement in the luminescence performance of warm red light-emitting LiSrVO4:Pr3+ phosphors

Warm red-emitting praseodymium-doped LiSrVO₄ phosphors were synthesized via solid-state reaction. The phase formation was verified using an X-ray diffraction study and the morphology was investigated using a scanning electron microscope study. The LiSrVO₄:Pr³⁺ phosphors emitted red light when exposed to ultraviolet light, indicating their possibility for use in warm white light-emitting diodes (WLEDs). Furthermore, the effect of charge compensators on the luminescence characteristics was addressed. The decay time was investigated using time-resolved photoluminescence. Furthermore, thermal quenching was analyzed through temperature-dependent photoluminescence spectra. Their sensitivity was calculated using temperature-dependent decay time analysis. The colour purity of the emitted light could be measured by photometric analysis. This comprehensive investigation provides a thorough understanding of the luminescence properties of phosphors for WLED applications.