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Electrophoresis Technique (electrophoresis + technique)
Selected AbstractsUniversal method for synthesis of artificial gel antibodies by the imprinting approach combined with a unique electrophoresis technique for detection of minute structural differences of proteins, viruses, and cells (bacteria).ELECTROPHORESIS, Issue 23 2006III: Gel antibodies against cells (bacteria) Abstract Artificial antibodies in the form of gel granules were synthesized from the monomers acrylamide and N,N'-methylenebisacrylamide by the imprinting method in the presence of Echerichia coli bacteria as template. The electrophoretic migration velocities of the gel antibodies (i),saturated with the antigen (Escherichia,coli MRE-600), (ii),freed of the antigen, and (iii),resaturated with bacteria, were determinated by electrophoresis in a rotating narrow-bore tube of 245,mm length and the 2.5 and 9.6,mm inner and outer diameters, respectively. Removal of bacteria from the gel antibodies was made by treatment with enzymes, followed by washing with SDS and buffer. Gel granules becoming charged by adsorption of bacteria move in an electrical field. We obtained a significant selectivity of gel antibodies for E.,coli MRE-600, since the granules did not interact with Lactococcus lactis; and when E.,coli BL21 bacteria were added to the gels selective for E.,coli MRE-600, a significant difference in the migration rate of the complexes formed with the two strains was observed indicating the ability of differentiation between the two strains. The gel antibodies can be used repeatedly. The new imprinting method for the synthesis of artificial gel antibodies against bioparticles described herein, and the classical electrophoretic analysis technique employed, thus represent , when combined , a new approach to distinguish between different types and strains of bacteria. The application area can certainly be extended to cover other classes of cells. [source] A simple polyacrylamide gel electrophoresis procedure for separation of polyamidoamine dendrimersELECTROPHORESIS, Issue 16 2003Ajit Sharma Abstract A simple, inexpensive, and rapid electrophoresis technique was developed for use as a routine tool for evaluating purity of polyamidoamine (PAMAM) dendrimers. A variety of factors influencing migration of generations 0,7 dendrimers on nongradient polyacrylamide gels were evaluated. The low generation dendrimers were found to be very sensitive to diffusion during or after electrophoresis. The proposed method incorporates steps that minimize diffusion, in order to obtain improved resolution and sensitivity, especially for the lower-molecular-weight dendrimers. This was accomplished by inclusion of a dendrimer fixation step with glutaraldehyde and performing the electrophoresis separation, fixation, staining, and destaining at 4°C. PAMAM dendrimer separation was studied under basic and acidic conditions. Electrophoresis under acidic conditions gave increased resolution and sensitivity over separation at alkaline pH. Oligomers and trailing generations could be clearly separated and visualized under these conditions. The smallest PAMAM dendrimer, generation 0, was visible at 1.5 ,g under the optimized acidic conditions. With slight modifications, this technique should be applicable to separation of other water-soluble dendrimers. [source] Genotypic characterization of Porphyromonas gingivalis isolated from subgingival plaque and blood sample in positive bacteremia subjects with periodontitisJOURNAL OF CLINICAL PERIODONTOLOGY, Issue 9 2008P. Juliana Pérez-Chaparro Abstract Aim: The objective of this study was to investigate clonal relationship among Porphyromonas gingivalis isolated from subgingival plaque and blood samples in positive transient bacteremia subjects with periodontitis. Material and Methods: Unrelated patients with general chronic periodontitis or general aggressive periodontitis requiring scaling and root planing (SRP) were included in the study. Genotyping of each isolate was performed using pulsed field gel electrophoresis technique. Genetic relatedness of strains isolated within an individual or between different patients was determined by dendogram analysis. Results: Following SRP, from 16 patients, seven patients showed positive P. gingivalis bacteremia and nine were negative. Thirty-two strains were isolated from subgingival plaque and blood samples before and during induced transient bacteremia. The majority of the patients harboured one clonal type. Two patients showed different clones in plaque and blood samples suggesting that more than one clone can be found in subgingival plaque. P. gingivalis isolates from periodontitis patients after transient bacteremia following SRP, revealed a high heterogeneity among isolates. Conclusion: In 6/16 subjects the same P. gingivalis isolate was found in the blood and in oral cavity. P. gingivalis heterogeneity suggests no association of a unique clonal type with transient bacteremia. [source] Analytical aspects of pharmaceutical grade chondroitin sulfatesJOURNAL OF PHARMACEUTICAL SCIENCES, Issue 12 2007Nicola Volpi Abstract Chondroitin sulfate is a very heterogeneous polysaccharide in terms of relative molecular mass, charge density, chemical properties, biological and pharmacological activities. It is actually recommended by EULAR as a symptomatic slow acting drug (SYSADOA) in Europe in the treatment of knee osteoarthritis based on meta-analysis of numerous clinical studies. Chondroitin sulfate is also utilized as a nutraceutical in dietary supplements mainly in the United States. On the other hand, chondroitin sulfate is derived from animal sources by extraction and purification processes. As a consequence, source material, manufacturing processes, the presence of contaminants, and many other factors contribute to the overall biological and pharmacological actions of these agents. The aim of this review is to evaluate new possible more specific analytical approaches to the determination of the origin and purity of chondroitin sulfate preparations for pharmaceutical application and in nutraceuticals, such as the evaluation of the molecular mass values, the constituent disaccharides, and the specific and sensitive agarose-gel electrophoresis technique. Furthermore, a critical evaluation is presented, together with a discussion of the limits of these analytical approaches. Finally, the necessity for reference standards having high specificity, purity and well-known physico-chemical properties useful for accurate and reproducible quantitative analyses will be discussed. © 2007 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci 96: 3168,3180, 2007 [source] Advances in protein turnover analysis at the global level and biological insightsMASS SPECTROMETRY REVIEWS, Issue 5 2010Qingbo Li Abstract The concept of a dynamic state of body constituents, a precursor of the modern term of proteome dynamics, was conceived over a century ago. But, not until recently can we examine the dynamics of individual "constituents" for example, proteins at a truly global level. The path of advancement in our understanding of protein turnover at the global level is marked by the introduction of some key technological innovations. These methods include the isotopic tracer technique in the 1930s, the two-dimensional gel electrophoresis technique in the 1970s, the sector mass spectrometer that could analyze isotopomers of peptides in the early 1990s, the 2D gel/MALDI-TOF proteomics technology in the late 1990s, the booming liquid chromatography/mass spectrometry proteomics technology in this decade, and the recently emerging protein-tagging approaches that offer single-cell resolution for protein turnover measurements. The long-standing inquiry raised in the 1950s about the existence of a dynamic state in different organisms at different physiological conditions can now be answered with an individual "constituent" resolution on a truly global scale. Now it appears that protein degradation is not necessarily an end to the protein function. Rather, it can be the start of a new function because protein degradation clears the way for the action of other proteins. Protein turnover participates in a multi-layer complex regulatory network and shares equal importance with gene transcription and protein translation. The advances in technologies for protein turnover analysis and the improved understanding of the biological role of protein turnover will likely help to solve some long-standing biomedical problems such as the tuberculosis disease that at the present day still affects one-third of the world population. © 2009 Wiley Periodicals, Inc., Mass Spec Rev 29:717,736, 2010 [source] Copper(II),Girard's T complex as a promising anti-tumor agentAPPLIED ORGANOMETALLIC CHEMISTRY, Issue 6 2010A. M. A. El-Sokkary Abstract A copper(II) complex was evaluated for its anti-tumor activity. Firstly, electrophoretic studies were applied on the complex. These studies revealed the binding of the complex to calf thymus DNA, leading to a delay in electrophoretic mobility of the DNA molecule. Secondly, spectroscopic data pointed out that the ,max of DNA was shifted to a longer wavelength, which was accompanid by a hyperchromic shift. Moreover, the ,max of copper(II) complex was shifted to a shorter wavelength. The favorable reaction conditions between the DNA molecule and the copper(II) complex were studied. Thirdly, The effects of the ligand and the Cu(II) ion were tested separately on the DNA molecule by electrophoresis technique. Furthermore, the fluorescence quenching of DNA bound ethidium ion by Cu(II)-Girard's T complex was noticed. The IR spectral data of DNA before and after the reaction with the copper(II) complex indicated that the interaction takes place through the carbonyl group of DNA nucleobases. Finally, a significant increase in the mean survival of EAC (Ehrlich ascites carcinoma) tumor-bearing mice was observed when treated with the copper(II) complex. The tumor volume was also significantly reduced (p < 0.0001). Electrophoretic studies showed that the DNA pattern extracted from EAC cells of tumor-bearing mice was affected after treatment with the copper(II) complex. Flow cytometric studies showed that this complex may be taken into consideration in seeking novel anti-tumor agents. Copyright © 2010 John Wiley & Sons, Ltd. [source] Mutational screening of the cationic trypsinogen gene in a large cohort of subjects with idiopathic chronic pancreatitisCLINICAL GENETICS, Issue 3 2001Jm Chen Several missense mutations, including R122H, N29I, K23R, A16V and D22G, in the cationic trypsinogen gene (PRSS1), have been associated with certain forms of hereditary pancreatitis (HP). Their occurrence in the idiopathic chronic pancreatitis (ICP) and whether novel mutations could be identified in PRSS1 remain to be further evaluated. These were addressed by the mutational screening of the entire coding sequence and the intronic/exonic boundaries of the PRSS1 gene in 221 ICP subjects, using a previously established denaturing gradient gel electrophoresis technique. Among the known PRSS1 mutations, only the R122H was detected in a single subject and the A16V in two subjects in the cohort, strengthening that HP-associated PRSS1 mutations are rare in ICP. Additional missense mutations, including P36R, E79K, G83E, K92N and V123M, were identified once separately. By analogy with the known PRSS1 mutations, predisposition to pancreatitis by some of them, particularly the V123M autolysis cleavage site mutation, is suspected. Functional analysis is expected to clarify their possible medical consequences. [source] | |||||||||||||