An approach to remove alpha amylase for proteomic analysis of low abundance biomarkers in human saliva

Proteomic characterization of human whole saliva for the identification of disease-specific biomarkers is guaranteed to be an easy-to-use and powerful diagnostic tool for defining the onset, progression and prognosis of human systemic diseases and, in particular, oral diseases. The high abundance of proteins, mainly alpha amylase, hampers the detection of low abundant proteins appearing in the disease state and therefore should be removed. In the present study a 2-DE was used to analyze human whole saliva following the removal of alpha amylase by affinity adsorption to potato starch. After alpha amylase removal whole saliva was analyzed by SDS-PAGE showing at least sixfold removal efficiency and by an alpha amylase activity assay showing 97% reduced activity. MS identification of the captured alpha amylase after elution demonstrated specific removal; 2-DE analysis showed the selective removal of alpha amylase and consequently increased gel resolution. MS identification of protein spots in the 60 kDa area revealed 15 proteins, which were masked before alpha amylase removal. In conclusion, treatment of human whole saliva with an alpha amylase removal device increases gel resolution and enables a higher protein sample for analysis.     

Nanoemulsion drug delivery by ketene based polyester synthesized using electron rich carbon/silica composite surface

A new carrier matrix for nanoemulsion drug delivery was synthesized from glycine as the raw material, using mesoporous/microporous electron rich carbon-silica composite surface (MAC(800)). MAC(800) was prepared from rice husk in two-stage carbonization. The surface area, pore volume, and pore size distribution of MAC(800) were measured, using nitrogen adsorption isotherms at 77K. The unpaired electron density of MAC(800) was measured in electron spin resonance spectroscopy (ESR), using TEMPOL (4-hydroxy-2,2,6,6-tetramethyl piperidine-1-oxyl) as the reference spin probe. Glycine was converted into ketene at the surface of MAC(800), which further underwent radical polymerization to form a low molecular weight ketene polymer (LMKP) of ester structure. The structure and the properties of LMKP were confirmed through (13)C, (1)H and DEPT nuclear magnetic resonance (NMR) spectroscopy, attenuated total reflectance-Fourier transform infrared spectroscopy (ATR-FTIR) and size exclusion chromatography (SEC). The two hydrophilic drugs namely ciprofloxacin hydrochloride (CPH) and gentamicin sulphate (GS) were chosen for the nanoemulsion preparation and characterization. They were characterized for morphology, interaction of drugs with the polymer and their crystallinity, using HR-TEM, DSC and XRD, respectively. The encapsulation efficiency of the LMKP towards the drugs ciprofloxacin hydrochloride and gentamicin sulphate were 26% and 12%, respectively. The dissolution studies of the nanoemulsion were carried out for the pH 6.5, 7.4 and 8.0. The cytocompatibility studies were done for LMKP as well as nanoemulsion using Hep2 epithelial cells.     

Miscellaneous PEDOT:PTS (polythiophenesulfonyl chloride) based conductive binder for silicon anodes in lithium ion batteries

Lithium ion batteries which are an energy storage system have increasing attention owing to suitability and advantages for many applications. Although it has ideal specifications, the capacity properties still have to be developed. In this study, the electrical conductivity of the anode was increased by using a conductive polymer binder and the active material content of the anode was also enhanced without adding carbon additives. Silicon based anodes were manufactured by using poly(3,4-ethylenedioxythiophene)/polystyrene sulfonate (PEDOT:PSS) and poly(3,4-ethylenedioxythiophene)/polythiophenesulfonyl chloride (PEDOT:PTS) conductive polymer binders. Si/PEDOT:PTS anode showed about 2000 mAh/g specific capacities after 60 cycles with decreasing impedance.     

Chemical Synthesis of Bioactive Proteins

Nature has developed a plethora of protein machinery to operate and maintain nearly every task of cellular life. These processes are tightly regulated via post-expression modifications—transformations that modulate intracellular protein synthesis, folding, and activation. Methods to prepare homogeneously and precisely modified proteins are essential to probe their function and design new bioactive modalities. Synthetic chemistry has contributed remarkably to protein science by allowing the preparation of novel biomacromolecules that are often challenging or impractical to prepare via common biological means. The ability to chemically build and precisely modify proteins has enabled the production of new molecules with novel physicochemical properties and programmed activity for biomedical research, diagnostic, and therapeutic applications. This minireview summarizes recent developments in chemical protein synthesis to produce bioactive proteins, with emphasis on novel analogs with promising in vitro and in vivo activity.     

Effect of sonication and freezing-thawing on the aggregate size and dynamic surface tension of aqueous DPPC dispersions

The effect of sonication and freezing-thawing on the aggregate size and dynamic surface tension of aqueous dipalmitoylphosphatidylcholine (DPPC) dispersions was studied by cryogenic-transmission electron microscopy (cryo-TEM), dynamic light scattering (DLS), UV-vis spectroturbidimetry, and surface tensiometry. When 1000 ppm (0.1 wt%) DPPC dispersions were prepared with a certain protocol, including extensive sonication, they contained mostly frozen vesicles and were quite clear, transparent, and stable for at least 30 days. The average dispersed vesicles diameter was 80 nm in water and 90 nm in standard phosphate saline buffer. After a freeze-thaw cycle, this dispersion became turbid, and precipitates of coagulated vesicles were observed with large particles of average size of 1.5x10(3) nm. The vesicle coagulation is due to the local salt concentration increase during the freezing of water. This dispersion has much higher equilibrium and dynamic surface tension than those before freezing. When this freeze-thawed dispersion was subjected to a resonication at 55 degrees C, smaller vesicles with sizes of ca. 70 nm were produced, and a lower surface tension behavior was restored as before freezing. Similar behavior was observed at 30 ppm DPPC. These results indicate that the freeze-thaw cycle causes substantial aggregation and precipitation of the vesicles. These results have implications for designing efficient protocols of lipid dispersion preparation and lung surfactant replacement formulations in treating respiratory disease and for effective administration.     

菲啶和三苯基膦的Cu(Ⅰ)/Cu(Ⅱ)配合物的合成、表征和X射线晶体结构分析

以三苯基膦（PPh₃）为共配体,合成了2种新的铜（Ⅰ）和铜（Ⅱ）与菲啶（Phend）的配合物,其组成为[Cu（κ¹-Phend）₂Cl₂]（1）和[Cu₂（κ¹-Phend）₂（κ¹-PPh₃）₂（μ-Cl）₂]（2）。这些配合物的结构通过元素分析、摩尔电导率、FT-IR、UV-Vis和单晶X射线衍射进行了研究。典型配合物1的X射线衍射分析显示,Cu（Ⅱ）配位构型为扭曲的平面四方形,而双核配合物2的Cu（Ⅰ）中心为含μ-Cl^-离子的不规则四面体构型。FT-IR谱、元素分析以及UV-Vis谱证实了它们的成分、几何形状和配体相互作用。2种配合物的结构通过密度泛函理论（DFT）计算进行了优化,以解释电子光谱特性。     

Unitary periods,Hermitian forms and points on flag varieties

Let E be an imaginary quadratic extension of of class number one. We examine certain representation numbers associated to Hermitian forms over E , which involve counting integral points on flag varieties. G. Chinta is partially supported by the NSF and by a Humboldt Research Fellowship.     

Activation of p300 histone acetyltransferase by small molecules altering enzyme structure: probed by surface-enhanced Raman spectroscopy

Reversible acetylation of nucleosomal histones and nonhistone proteins play pivotal roles in the regulation of all the DNA templated phenomenon. Dysfunction of the enzymes involved in the acetylation/deacetylation leads to several diseases. Therefore, these enzymes are the targets for new generation therapeutics. Here, we report the synthesis of trifluoromethyl phenyl benzamides and their effect on histone acetyltransferase (HAT) activity of p300. One of these benzamides, CTPB (N-(4-chloro-3-trifluoromethyl-phenyl)-2-ethoxy-6-pentadecyl-benzamide), was discovered as a potent activator of the p300 HAT activity. We have found that pentadecyl hydrocarbon chain of CTPB is required to activate the HAT only under certain context. Furthermore, our results show that the relative position of -CF3 and -Cl in CTB (N-(4-chloro-3-trifluoromethyl-phenyl)-2-ethoxy-benzamide) is also very critical for the activation. Surface-enhanced Raman spectroscopy (SERS) of p300 and the HAT activator complexes evidently suggest that the activation of HAT activity is achieved by the alteration of p300 structure. Therefore, apart from elucidating the chemical basis for small molecule mediated activation of p300, this report also describes, for the first time, Raman spectroscopic analysis of the complexes of histone-modifying enzymes and their modulators, which may be highly useful for therapeutic applications.