Thermo- and pH-sensitive poly(vinylmethyl ether)/carboxymethylchitosan hydrogels crosslinked using electron beam irradiation or using glutaraldehyde as a crosslinker

BACKGROUND: Stimuli-sensitive or intelligent hydrogels have been investigated for many biomedical and pharmaceutical applications. Those hydrogels with dual sensitivity will have more extensive potential applications. The aim of the work presented was to prepare a series of thermo- and pH-sensitive hydrogels based on poly(vinylmethyl ether) (PVME) and carboxymethylchitosan (CMCS). The hydrogels were crosslinked using electron beam irradiation (EB) or using glutaraldehyde (GA) as a crosslinker at room temperature.

Spectroscopic investigation of the function of aqueous 2-hydroxyethylmethacrylate/glutaraldehyde solution as a dentin desensitizer

Fourier-transform (FT)-Raman and -infrared (IR) spectroscopy were employed to investigate the function of the aqueous 2-hydroxyethylmethacrylate/glutaraldehyde solution (Gluma) as a desensitizer. 2-Hydroxyethylmethacrylate (HEMA), glutaraldehyde (GA), and the mixture of HEMA/GA (i.e. Gluma) were used to interact with dentin, collagen, hydroxyapatite (HAP), and bovine serum albumin (BSA) individually. All the interactions were monitored by an FT-Raman spectrometer. FT-IR spectroscopy was also used in this study. The results show that HEMA could be absorbed by dentin and collagen; GA could cross-link collagen and BSA; and when BSA was added to Gluma, polymerization of HEMA occurred. The results suggest that Gluma acts as a desensitizer whereby, first, GA reacts with part of the serum albumin in dentinal fluid, which induces a precipitation of serum albumin, then, second, a reaction of GA with serum albumin induces polymerization of HEMA. The function of Gluma as a desensitizer to block dentinal tubules occurs via these two reactions.

Comparative studies on the efficiency of energy transfer of two R-phycoerythrin-C-phycocyanin conjugates constructed from sodium periodate and glutaraldehyde respectively

The C-phycocyanin and the R-phycoerythrin were purified from the blue-green alga Spirulina platensis and red alga Polysiphonia urceolata respectively. Both sodium periodate and glutaraldehyde are effective coupling agents being capable of constructing the R-phycoerythrin-C-phycocyanin conjugate, which was also called phycobiliproteins energy transfer model. The two artificial conjugates constructed with different methods were purified by Sephadex G-200 chromatography respectively. Spectra analysis indicated that energy transfer occurred in the two conjugates. The conjugate with sodium periodate had the higher efficiency of energy transfer than that with glutaraldehyde conjugate.

The stabilization effect of glutaraldehyde on the Spirulina plateusis phycobilisomes

The spectral properties of the glutaraldehyde-treated phycobilisomes were studied. The results showed that glutaraldehyde was effective in preventing phycobilisomes from dilution-induced dissociation and preserving the intra-phycobilisomes energy transfer.

Improvement and observation of immunoelectron microscopic method for the localization of frog Rana grylio virus (RGV) in infected fish cells

In this paper, to understand the roles of amorphous structures which were observed within the viromatrix of Rana grylio virus (RGV), an improved immunoelectron microscopy (IEM) method was developed to detect the localization of RGV in carp Epithelipma papulosum cyprinid (EPC) cells. Infected EPC cells were fixed with 4% paraformaldehyde-0.25% glutaraldehyde mixture, dehydrated completely, and embedded in LR White resin. This method allowed good ultrastructural preservation and specific labeling with anti-RGV antibodies. The results of IEM showed that colloidal gold mainly bound to the capsids of viral particles at the stage of viral assembly, while during the viral maturation colloidal gold bound to the envelop of virions. In addition, within the viromatrix, the amorphous structures, including dense floccules, membranous materials and tubules, also had strong colloidal gold signals, revealing that those amorphous structures were participated in RGV assembly. In contrast, no significant gold labeling signals were obtained in negative controls. The present study not only provided further evidence that amorphous structures within the viromatrix were involved in the process of RGV assembly, but also developed an improved IEM method for studying the interaction between iridovirus and host cells. (C) 2006 Elsevier Ltd. All rights reserved.

真菌单宁酶发酵五倍子和有机溶剂中酶法合成没食子酸丙酯的研究

本文报告了丝状真菌单宁酶发酵五倍子及有机溶剂中酶法合成没食子酸丙酯的研究。利用单宁和/或五倍子诱导丝状真菌产生单宁 酶的原理，借助二级发酵程序，对从天然源得到的75株菌进行了生物转化实验研究。选择出既能水解单宁或五倍子成没食子酸，又 能把没食子酸和丙醇合成没食子酸丙酯，而且生物催化活性都较高的1株菌，这株菌经初步鉴定为黑曲霉(Aspergillus niger No.17)。随后对它开展了产酶条件和参数优化实验，得出了最佳培养条件。立足于参数优化实验方案的基础上，经由液体培养发酵 制备单宁酶制剂，并把该酶通过化学手段共价结合到一种新型载体—聚乙烯醇和戊二醛反应生成的缩醛上，制备得到固定化单宁酶 。这种固定化生物催化剂在两种有机介质体系中都具有逆向催化合成没食子酸丙酯的能力。最后建立起来一条有效可行的微生物酶 法制备没食子酸的技术途径，没食子酸产率达到70%。对这种物质进行元素 分析：含C,49.45%;含H,3.63%。它的熔点为237℃～243 ℃，三种溶剂系统的TLC均只给出一个斑点。这些数据都与标准品一致。有机溶剂中酶法合成没食子酸丙酯的技术途径已经建立。 水溶性单宁酶在潜溶剂体系中也能催化上述酯化反应，反应混合物中的PG浓度为16.4mmol/L，制备薄层被用于分离反应混合物所含 的PG，这种产物被红外、质谱及三种溶剂系统的TLC等方法鉴定，确证为目标产物。在这一学位论文的实验研究过程中，还包括一 些生化分析方法的建立和应用，这些方法用于鉴定底物和产物及测定它们的浓度，其内容主要包括TLC定性/半定量分析、元素分析 、质谱、红外等手段的综合运用。本工作为开发我国特有的天然产物资源—五倍子的生物化工加工技术及非水相生物催化技术的开 发，提供了有用的基础数据资料，具有应用基础研究工作的重要性。In this thesis, the studies on the fermentation of Chinese gallotannin by filamentous fungi with tannase activity and enzymatic synthesis of propyl gallate(PG) in organic solvents were described through these biocatalysts. Based on the principles of induction enzyme, the tannase produced from filamentous fungi by adding tannic acid(TA) and/or Chinese gallotannin into media was investigated, and the screening experiments of bioconversion were done with 75 strains by means of a two-stage fermentation procedure. These strains were isolated with the enrichment culture technique from natural sources. Hence we selected one strain (Aspergillus niger No.17) that can not only catalyze the hydrolyses of TA and/or Chinese gallotannin into gallic acid(GA) in the liquid cultures, but also be used to synthesize PG from propanol and GA in the non-aqueous media. At the same time both of its biocatalytical activities were higher. This strain was calssified to be Aspergillus niger by the primary identification. Then optimum conditions for production of the tannase and its parameters were examined. In this way, one set of optimum culture conditions was selected. Making use of the optimum proposal, the tanase was prepared through a liquid fermentation procedure. The enzyme was convalently coupled to a new type of carrier which was made chemically from polyvinyl alcohol(PVA)and glutaraldehyde. The immobilized enzymes were able to synthesize PG reversely in two organic media. Finally, an effective enzymatic technique for production of GA was developed. The yield of GA products was up to 70%。Element analysis for this substance: calce: C, 49.42%; H, 3.56%; found: C, 49.45%, H, 3.63%. Its melting point was 237℃～ 243℃ and TLCs on three solvent systems gave only one spot respectively. These data were identical with theauthentic GA. The enzymatic synthesis of PG in organic solvents was extablished with reverse route of tannase catalytical hydrolysis. Aqueous enzyme perparation also catalyzed above esterification in a buffer system. The PG concentration in the reaction mixture was 16.4mmol/L. The reparative-scale TLC was used to isolate PG from the reaction mixture. This product separated was identified by IR, MS and TLC on three solvent systems. In this study of thesis, some biochemical analytical mehtods were developed and used to identify substrates and products, and to determinate their concentration. These methods, including TLC qualitative/half quantitative analysis, element analysis, MS, IR and so on, were useful, available and performable. This work provided basic data and information for developing the biochemical engineering and bio-processing of Chinese gallotannin-a special natural resource in China and the non-aqueous phase biocatalysis. Thus, this study possesses importance in the applied and basic research work.

Repeatedly usable immobilized pH gradient in a monolithic capillary column

Glycidyl methacrylate (GMA) and ethylene dimethacrylate (EDMA) were used to synthesize a monolithic capillary column containing reactive epoxy groups. Glutaraldehyde was introduced and linked to the monolith after a process of amination. An aqueous solution of commercial carrier ampholytes (CAs, Ampholine) was focused in such a polymer column. The primary amino groups of CAs reacted with glutaraldehyde along the capillary. CAs were immobilized at different positions in the column according to their isoelectric points (pl), resulting in a monolithic immobilized pH gradient (M-IPG). Isoelectric focusing (IEF) was performed without CAs in such an M-IPG column. Due to the covalent attachment of the CAs this M-IPG can be repeatedly used after its preparation. Good stability, linearity, and reproducibility were obtained.

Biosorption of Cd2+, Cu2+, Ni2+ and Zn2+ ions from aqueous solutions by pretreated biomass of brown algae

In this paper, marine brown algae Laminaria japonica was chemically modified by crosslinking with epichlorohydrin (EC1 and EC2), or oxidizing by potassium permanganate (PC), or crosslinking with glutaraldehyde (GA), or only washed by distilled water (DW). They were used for equilibrium sorption uptake studies with Cd2+, Cu2+, Ni2+ and Zn2+.

Electrochemical and electrochemiluminescence study of Ru(bpy)(3)(2+)-doped silica nanoparticles with covalently grafted biomacromolecules

Spherical Ru(bpy)(3)(2+)-doped silica (RuSi) nanoparticles were prepared via a water-in-oil microemulsion approach. The electrochemical and electrochemiluminescent properties of the RuSi nanoparticles immobilized on an indium tin oxide (ITO) electrode were investigated. Further, electrochemiluminescence (ECL) of the RuSi nanoparticles with covalently coated biomacromolecules was studied. By covalent cross-linking with glutaraldehyde, gamma-(aminopropyl) triethoxysilane (APTES)-pretreated RuSi nanoparticles were coupled with different concentrations of bovine serum albumin (BSA), hemoglobin, and myoglobin, respectively.

Preparation and application of single polyelectrolyte microcapsules possessing tunable autofluorescent properties

Autofluorescent single polyelectrolyte microcapsules, exemplified by poly-L-lysine (PLL), have been prepared through glutaraldehyde-mediated covalent layer-by-layer (LbL) assembly and subsequent core removal. CaCO3 microparticles were used as template cores for the LbL deposition and removed by treatment of ethylenediamine tetraacetic acid disodium salt (EDTA). The prepared microcapsules, without conjugating an exterior fluorochrome, exhibited evenly distributed fluorescence.

A sensitive impedimetric thrombin aptasensor based on polyamidoamine dendrimer

A label-free and highly sensitive impedimetric aptasensor based on a polyamidoamine dendrimer modified gold electrode was developed for the determination of thrombin. Amino-terminated polyamidoamine dendrimer was firstly covalently attached to the cysteine functionalized gold electrode through glutaraldehyde coupling. Subsequently, the dendrimer was activated with glutaraldehyde, and amino-modified thrombin aptamer probe was immobilized onto the activated dendrimer monolayer film. The layer-by-layer assembly process was traced by surface plasmon resonance and electrochemical impedance spectroscopy.

Biosorption of Cd2+, Cu2+, Ni2+ and Zn2+ ions from aqueous solutions by pretreated biomass of brown algae

In this paper, marine brown algae Laminaria japonica was chemically modified by crosslinking with epichlorohydrin (EC1 and EC2), or oxidizing by potassium permanganate (PC), or crosslinking with glutaraldehyde (GA), or only washed by distilled water (DW). They were used for equilibrium sorption uptake studies with Cd2+, Cu2+, Ni2+ and Zn2+. The experimental data have been analyzed using Langmuir, Freundlich and Redlich-Peterson isotherms. The results showed that the biosorption equilibrium was well described by both the Langmuir and Redlich-Peterson isotherms.

Fabrication and characterization of microstructured and pH sensitive interpenetrating networks hydrogel films and application in drug delivery field

Novel microstructured and pH sensitive poly(acryliac acid-co-2-hydroxyethyl methacrylate)/poly(vinyl alcohol) (P(AA-co-HEMA)/PVA) interpenetrating network (IPN) hydrogel films were prepared by radical precipitation copolymerization and sequential IPN technology. The first P(AA-co-HEMA) network was synthesized in the present of IPN aqueous solution by radical initiating, then followed by condensation reaction (Glutaraldehyde as crosslinking agent) within the resultant latex, it formed multiple IPN microstructured hydrogel film. The film samples were characterized by IR, SEM and DSC. Swelling and deswelling behaviors and mechanical property showed the novel multiple IPN nanostuctured film had rapid response and good mechanical property. The IPN films were studied as controlled drug delivery material in different pH buffer solution using cationic compound, crystal violet as a model drug.

The modification of screen-printed carbon electrodes with amino group and its application to construct a H2O2 biosensor

Electrooxidation of thionine on screen-printed carbon electrode gives rise to the modification of the surface with amino groups for the covalent immobilization of enzymes such as horseradish peroxidase (HRP). The biosensor was constructed using multilayer enzymes which covalently immobilized onto the surface of amino groups modified screen-printed carbon electrode using glutaraldehyde as a bifunctional reagent. The multilayer assemble of HRP has been characterized with the cyclic voltammetry and the faradaic impedance spectroscopy. The H2O2 biosensor exhibited a fast response (2 s) and low detection limit (0.5 muM).

Composite film of carbon nanotubes and chitosan for preparation of amperometric hydrogen peroxide biosensor

A new amperometric biosensor for hydrogen peroxide was developed based on cross-linking horseradish peroxidase (HRP) by glutaraldehyde with multiwall carbon nanotubes/chitosan (MWNTs/chitosan) composite film coated on a glassy carbon electrode. MWNTs were firstly dissolved in a chitosan solution. Then the morphology of MWNTs/chitosan composite film was characterized by field-emission scanning electron microscopy. The results showed that MWNTs were well soluble in chitosan and robust films could be formed on the surface. HRP was cross-linked by glutaraldehyde with MWNTs/chitosan film to prepare a hydrogen peroxide biosensor. The enzyme electrode exhibited excellent electrocatalytic activity and rapid response for H2O2 in the absence of a mediator. The linear range of detection towards H2O2 (applied potential: -0.2 V) was from 1.67 x 10(-5) to 7.40 x 10(-4) M with correction coefficient of 0.998. The biosensor had good repeatability and stability for the determination of H2O2. There were no interferences from ascorbic acid, glucose, citrate acid and lactic acid.