SUMOylation in Control of Accurate Chromosome Segregation during Mitosis

Posttranslational protein modification by small ubiquitin-related modifier (SUMO) has emerged as an important regulatory mechanism for chromosome segregation during mitosis. This review focuses on how SUMOylation regulates the centromere and kinetochore activities to achieve accurate chromosome segregation during mitosis. Kinetochores are assembled on the specialized chromatin domains called centromeres and serve as the sites for attaching spindle microtubule to segregate sister chromatids to daughter cells. Many proteins associated with mitotic centromeres and kinetochores have been recently found to be modified by SUMO. Although we are still at the early stage of elucidating how SUMOylation controls chromosome segregation during mitosis, a substantial progress has been achieved over the past decade. Furthermore, a major theme that has emerged from the recent studies of SUMOylation in mitosis is that both SUMO conjugation and deconjugation are critical for kinetochore assembly and disassembly. Lastly, we propose a model that SUMOylation coordinates multiple centromere and kinetochore activities to ensure accurate chromosome segregation.     

表面接枝季铵盐型高分子材料抗菌过程的特性研究

考察了一种表面接枝季铵盐型高分子材料对水溶液中大肠杆菌的抗菌过程特性,发现此材料可以在短时间内迅速降低溶液中的大肠杆菌的活菌量.用菌体耗氧测定法与扫描电镜观察法研究了其抗菌过程.结果表明,其抗菌过程由吸附和杀灭两个步骤构成,其中吸附过程由纤维与大肠杆菌表面静电作用所控制,是一个快速过程,而杀灭过程则伴随着细胞壁在纤维表面的溶解,是一个慢过程.菌体耗氧测定法与扫描电镜观察法所获得的结果具有一致性,并可广泛应用于研究抗菌纤维的抗菌作用机理及过程特性.     

A Pyranoxanthone as a Potent Antimitotic and Sensitizer of Cancer Cells to Low Doses of Paclitaxel

Microtubule-targeting agents (MTAs) remain a gold standard for the treatment of several cancer types. By interfering with microtubules dynamic, MTAs induce a mitotic arrest followed by cell death. This antimitotic activity of MTAs is dependent on the spindle assembly checkpoint (SAC), which monitors the integrity of the mitotic spindle and proper chromosome attachments to microtubules in order to ensure accurate chromosome segregation and timely anaphase onset. However, the cytotoxic activity of MTAs is restrained by drug resistance and/or toxicities, and had motivated the search for new compounds and/or alternative therapeutic strategies. Here, we describe the synthesis and mechanism of action of the xanthone derivative pyranoxanthone 2 that exhibits a potent anti-growth activity against cancer cells. We found that cancer cells treated with the pyranoxanthone 2 exhibited persistent defects in chromosome congression during mitosis that were not corrected over time, which induced a prolonged SAC-dependent mitotic arrest followed by massive apoptosis. Importantly, pyranoxanthone 2 was able to potentiate apoptosis of cancer cells treated with nanomolar concentrations of paclitaxel. Our data identified the potential of the pyranoxanthone 2 as a new potent antimitotic with promising antitumor potential, either alone or in combination regimens.     

Chromosome protein framework from proteome analysis of isolated human metaphase chromosomes

We have presented a structural model of the chromosome based on its constituent proteins. Development of a method of mass isolation for intact human metaphase chromosomes and proteome analysis by mass spectrometry of the isolated chromosomal proteins enabled us to develop a four-layer structural model of human metaphase chromosomes. The model consists of four layers, each with different chromosomal protein sets, i.e., chromosome coating proteins (CCPs), chromosome peripheral proteins (CPPs), chromosome structural proteins (CSPs), and chromosome fibrous proteins (CFPs). More than 200 identified proteins have been classified and assigned to the four layers with each layer occupying a distinct region of the chromosome. CCPs are localized at the most outer regions of the chromosomes and they attach to the regions tentatively and occasionally. CCPs include mostly mitochondrial and cytoplasmic proteins, e.g., 70 kDa heat shock protein 9B and Hsp60. CPPs are also localized at the peripheral regions of the chromosomes, but as the essential part of the chromosomes. CPPs include nucleolin, lamin A/C, fibrillarin, etc. CSPs are the primary chromosomal structure proteins, and include topoisomerase IIalpha, condensin subunits, histones, etc. CFPs have a fibrous nature, e.g., beta-actin, vimentin, myosin II, tublin, etc. A data set of these proteins, which we developed, contains essential chromosome proteins with classified information based on this four-layer model and presents useful leads for further studies on chromosomal structure and function.     

Thermal decomposition of crystalline Ni(II)-Cr(III) layered double hydroxide: a structural study of the segregation process

A structural study of the thermal evolution of Ni(0.69)Cr(0.31)(OH)(2)(CO(3))(0.155) x nH(2)O into NiO and tetragonal NiCr(2)O(4) is reported. The characteristic structural parameters of the two coexisting crystalline phases, as well as their relative abundance, were determined by Rietveld refinement of powder x-ray diffraction (PXRD) patterns. The results of the simulations allowed us to elucidate the mechanism of the demixing process of the oxides. It is demonstrated that nucleation of a metastable nickel chromite within the common oxygen framework of the parent Cr(III)-doped bunsenite is the initial step of the cationic redistribution. The role that trivalent cations play in the segregation of crystalline spinels is also discussed.     

Regulation of glutamine synthetase by enzyme catalyzed structural modification

A highly sophisticated mechanism for the regulation of glutamine metabolism in Escherichia coli involves the modification of the enzyme glutamine synthetase by the covalent attachment and release of adenylyl residues. Separate enzymes catalyze the adenylylation and deadenylylation reactions, and these enzymes are reciprocally controlled by the state of nitrogen nutrition of the cell. The resultant structural alterations in glutamine synthetase enable it to change its catalytic potential greatly in response to varying cellular demands, in order to maintain nitrogen homeostasis.     

Morphology of microphase separated domains in rod-coil copolymer melts

We investigate the morphology of microphase separated domains in diblock copolymers where each chain consists of a stiff rod block and a flexible coil block. A simplified phenomenological model system is introduced, which is coarse-grained in terms of the local concentration difference between the two blocks and the local director field of the rod part. Computer simulations of this set of time-evolution equations in two dimensions show in the weak segregation regime that the elastic energy in the rod-block rich domains affects drastically the structures of microphase separated domains. A coil-to-rod transition is incorporated into the model system to examine the elastic and anisotropic effects. The effects of the external electric field are also investigated to control the domain morphology.     

Acrylamide-Derived Ionome,Metabolic, and Cell Cycle Alterations Are Alleviated by Ascorbic Acid in the Fission Yeast

Acrylamide (AA), is a chemical with multiple industrial applications, however, it can be found in foods that are rich in carbohydrates. Due to its genotoxic and cytotoxic effects, AA has been classified as a potential carcinogen. With the use of spectrophotometry, ICP-OES, fluorescence spectroscopy, and microscopy cell growth, metabolic activity, apoptosis, ROS production, MDA formation, CAT and SOD activity, ionome balance, and chromosome segregation were determined in Schizosaccharomyces pombe. AA caused growth and metabolic activity retardation, enhanced ROS and MDA production, and modulated antioxidant enzyme activity. This led to damage to the cell homeostasis due to ionome balance disruption. Moreover, AA-induced oxidative stress caused alterations in the cell cycle regulation resulting in chromosome segregation errors, as 4.07% of cells displayed sister chromatid non-disjunction during mitosis. Ascorbic acid (AsA, Vitamin C), a strong natural antioxidant, was used to alleviate the negative impact of AA. Cell pre-treatment with AsA significantly improved AA impaired growth, and antioxidant capacity, and supported ionome balance maintenance mainly due to the promotion of calcium uptake. Chromosome missegregation was reduced to 1.79% (44% improvement) by AsA pre-incubation. Results of our multiapproach analyses suggest that AA-induced oxidative stress is the major cause of alteration to cell homeostasis and cell cycle regulation.     

Study of the bioeffects of CdTe quantum dots on Escherichia coli cells

Quantum dots (QDs) hold great potential for applications in nanomedicine, however, only a few studies investigate their toxic- and bio-effects. Using Escherichia coli (E. coli) cells as model, we found that CdTe QDs exhibited a dose-dependent inhibitory effect on cell growth by microcalorimetric technique and optical density (OD(600)). The growth rate constants (k) were determined, which showed that they were related to the concentration of QDs. The mechanism of cytotoxicity of QDs was also studied through the attenuated total reflection-fourier transform infrared (ATR-FTIR) spectra, fluorescence (FL) polarization, and scanning electron microscopy (SEM). It was clear that the cell out membrane was changed or damaged by the addition of QDs. Taken together, the results indicated that CdTe QDs have cytotoxic effects on E. coli cells, and this effects might attribute to the damaged structure of the cell out membrane, thus QDs and by-products (free radicals, reactive oxygen species (ROS), and free Cd(2+)) which might enter the cells.     

Modelling the kinetics of solute segregation to partial dislocations for isothermal microcalorimetric evaluations

A model is proposed to describe the kinetics of solute segregation to partial dislocations in solid solutions of cold-rolled alloys. The case when half edge and half screw dislocations are present is considered. The model gives account of the kinetic behaviour observed in a deformed Cu-19 at% Al alloy where two unknown processes could be assessed during calorimetric isothermal experiments. The faster process corresponds to segregation to screw dissociated dislocations while the slower one corresponds to segregation to edge dissociated dislocations. Experimental activation energies, larger for edge dislocations, are close to that for pipe diffusion along the partials corrected by pinner binding energy terms. It is also predicted that segregation occurs faster as the dislocation density is increased. A quantitative comparison of experimental results with model predictions is given.The authors would like to thank the Fondo de Desarrollo Científico y Tecnológico (FONDECYT) for financial support through Project 1950566, and the Institute de Investigaciones y Ensayes de Materiales (IDIEM), Facultad de Ciencias Físicas y Matemticas, Universidad de Chile, for the facilities provided for this research.     

Norvancomycin-capped silver nanoparticles: Synthesis and antibacterial activities against E. coli

The synthesis of norvancomycin (NVan)-capped silver nanoparticles (Ag@NVan) and their notable in vitro antibacterial activities against E. coli, a Gram-negative bacterial strain (GNB), are reported here. Mercaptoacetic acid-stabilized spherical silver nanoparticles with a diameter of 16±4 nm are prepared by a simple chemical reaction. The formation process of the silver nanoparticles is investigated by UV-visible (UV-vis) spectroscopy and transmission electron microscopy (TEM). NVan is then grafted to the terminal carboxyl of the mercaptoacetic acid in the presence of N-(3-Dimethylaminopropyl)-N'-ethylcarbodiimide hydrochloride (EDAC). The TEM images of single bacteria treated with Ag@NVan show that plenty of Ag@NVan aggregate in the cell wall of E. coli. A possible antibacterial mechanism is proposed that silver nanoparticles may help destroy the stability of the outer membrane of E. coli, which makes NVan easier to bind to the nether part of the peptidoglycan structure. The antibacterial activities of silver nanoparticles on their own, together with the rigid polyvalent interaction between Ag@NVan and cell wall, enables Ag@NVan to be an effective inhibitor of GNB. This kind of bionanocomposites might be used as novel bactericidal materials and we also provide an effective synthesis method for preparing functional bioconjugated nanoparticles here.     

Effect of Escherichia coliform on the biomineralization of calcium bilirubinate in mimic systems

The influence of Escherichia coliform (E. coli), especially the proteins it secretes, on the nucleation, growth and aggregation processes of calcium bilirubinate (CaBR) in different mimic systems, such as NaCl aqueous solution and model bile, is studied in this paper. The results show that in NaCl aqueous solution the morphology of calcium bilirubinate (CaBR) changes from amorphous sheet structure to a highly self-organized and highly self-replicated fractal structure that is accumulated by rhombic microcrystals after the addition of E. coli. In model bile with the existence of E. coli, CaBR also forms a fractal structure, but the fractal structure is staghorn-like. Meanwhile, the composition of the prepared CaBRs is nonstoichiometric, and the crystallization is greatly improved with the existence of E. coli. Besides, formation mechanism of CaBRs affected by E. coli is explored, showing that the proteins secreted by E. coli interact with the Ca(2+) ions to provide nucleation sites for CaBRs, and the conformation of the proteins becomes more ordered, resulting in the improvement of the crystalline of CaBRs. On the other hand, the interaction of proteins and the Ca(2+) ions also promote the aggregation state of the CaBRs.     

A model of the oxygen-evolving center of photosystem II predicted by structural refinement based on EXAFS simulations

A refined computational structural model of the oxygen-evolving complex (OEC) of photosystem II (PSII) is introduced. The model shows that the cuboidal core Mn3CaO4 with a "dangler" Mn ligated to a corner mu4-oxide ion is maximally consistent with the positioning of the amino acids around the metal cluster as characterized by XRD models and high-resolution spectroscopic data, including polarized EXAFS of oriented single crystals and isotropic EXAFS. It is, therefore, natural to expect that the proposed structural model should be particularly useful to establish the structure of the OEC, consistently with high-resolution spectroscopic data, and for elucidating the mechanism of water-splitting in PSII as described by the intermediate oxidation states of the EC along the catalytic cycle.     

Layer-by-layer assembly of nacre-like nanostructured composites with antimicrobial properties

In a recent report, we have presented the layer-by-layer (LBL) assembly of a biomimetic nanostructured composite from Na(+)-montmorillonite clay nanosheets and poly(diallylmethylammonium chloride) (Tang, Z.; Kotov, N.; Magonov, S.; Ozturk, B. Nat. Mater. 2003, 2, 413). The structure, deformation mechanism, and mechanical properties of the material are very similar to those of natural nacre and lamellar bones. This fact prompts further investigation of these composites as potential bone implants. LBL assembly affords preparation of multifunctional composites, and here we demonstrate that not only mechanical strength, but also antibacterial activity, can be introduced in these implantable materials by alternating clay layers with starch-stabilized silver nanoparticles. The resulting composite showed excellent structural stability with no detectable levels of silver lost over a 1 month period. Evaluation of the antibacterial properties showed almost complete growth inhibition of E. coli over an 18 h period. The amount of silver eluted from the LBL composite over a 1 month period was determined to be only 0.5-3.0 microg/L. This concentration of silver did not prevent the growth of the mammalian tissue cultures. The LBL composite has shown biocompatibility with the human osteoblast cell line.     

Membrane proteome in Escherichia coli probed by MS3 mass spectrometry: a preliminary report

Characterization of the membrane proteome is particularly intriguing since a better knowledge in this field might lead to new insights into the function of different membrane systems. Despite the biological relevance of surface proteins however, their characterization still remains a challenging task. Outer membrane proteins (OMPs) of Gram-negative bacteria are key molecules that interface the cell with the environment. Hence, surface proteins of Gram-negative bacteria contain proteins that might be good targets for drugs, antimicrobials or detection systems and they may become components of effective vaccines. In this respect, Escherichia coli has been chosen as a model organism for several structural and functional studies aimed at understanding the biophysical and biochemical organization of proteins in Gram-negative cell walls. Here we present first results for the identification of bacterial surface exposed proteins in E. coli K12 based on the use of dansyl chloride labelling coupled with bidimensional tandem mass spectrometry exploiting the advantage of precursor ion/MS3 scan modes. This procedure resulted in a promising, simple, and rapid strategy for the identification of membrane proteins in E. coli as model organism, thus avoiding time-consuming procedures based on two-dimensional liquid chromatography and electrophoresis. The proteins identified could be grouped into five major families: outer membrane (29 proteins), lipoproteins (6 proteins), transmembrane (43 proteins) families.     

Electrochemical evidence of self-substrate inhibition as functions regulation for cellobiose dehydrogenase from Phanerochaete chrysosporium

The reaction mechanism of cellobiose dehydrogenase (CDH) from Phanerochaete chrysosporium, adsorbed on graphite electrodes, was investigated by following its catalytic reaction with cellobiose registered in both direct and mediated electron transfer modes between the enzyme and the electrode. A wall-jet flow through amperometric cell housing the CDH-modified graphite electrode was connected to a single line flow injection system. In the present study, it is proven that cellobiose, at concentrations higher than 200 μM, competes for the reduced state of the FAD cofactor and it slows down the transfer of electrons to any 2e⁻/H⁺ acceptors or further to the heme cofactor, via the internal electron transfer pathway. Based on and proven by electrochemical results, a kinetic model of substrate inhibition is proposed and supported by the agreement between simulation of plots and experimental data. The implications of this kinetic model, called pseudo-ping-pong mechanism, on the possible functions CDH are also discussed. The enzyme exhibits catalytic activity also for lactose, but in contrast to cellobiose, this sugar does not inhibit the enzyme. This suggests that even if some other substrates are coincidentally oxidized by CDH, however, they do not trigger all the possible natural functions of the enzyme. In this respect, cellobiose is regarded as the natural substrate of CDH.     

Statistical Theory of Stable and Meta-stable Transition for Crystallization and Fusion of Homopolymers Ⅵ. Statistical Dynamics of Polymeric Crystallization by Molecular Segregation-Dependence of Crystal Growth Rate on the Grown Mechanism and Concentratio

First a short review on the dependence of crystal growth rate on the growth mechanism and concentration is present. Based on the structural model of micronucleus and crystal constituent chains, and the feature of statistical dynamics for polymeric crystallization by molecular segregation, a general method for characterizing number the growth rate and micro crystal constituent chains and the size growth rate for crystals was proposed. According to this method, a set of quantitative expressions for correlating the growth rate in number and size with the four types of growth (folding, extending and combination of folding and extending), the crystalline temperature and the crystalline concentration was derived. Then combined the concentration index is combined with the fraction of conformation for segments, a new correlation of the concentration index to the temperature of crystallization and the flexibility of polymeric chain is theoretically obtained. The dependences of the index on the different types of growth are also studied. Finally the relationships between the growth rate for crystals and the concentration of solution were verified by the experimental validating the predictions made by the theory.