Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society最新文献

{"title":"Cerebrospinal fluid protein variations in common to Alzheimer's disease and schizophrenia.","authors":"G Johnson,&nbsp;D Brane,&nbsp;W Block,&nbsp;D P van Kammen,&nbsp;J Gurklis,&nbsp;J L Peters,&nbsp;R J Wyatt,&nbsp;D G Kirch,&nbsp;H A Ghanbari,&nbsp;C R Merril","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Analysis of silver stained two-dimensional (2D) gels of cerebrospinal fluid (CSF) from 27 patients with schizophrenia (SCZ) and 10 patients with Alzheimer's disease (AD) revealed an increase in the relative amount of a polypeptide of 18,000M(r) and isoelectric point of 6.5 when compared to the appropriate controls. This protein was identified by its electrophoretic characteristics and by immune analysis of Western blots as an isoform of alpha-2 haptoglobin, provisionally identified as alpha-2FS haptoglobin. Alzheimer's disease versus control CSF samples showed a 6.8-fold increase in the percent mean density value of this haptoglobin isoform (n = 10 AD vs 11 control; P > 0.025) while a 4.4-fold increase was observed in the schizophrenic patients (n = 17 SCZ vs 10 control; P > 0.001). Two additional polypeptides (proteins '127' and '128') of 40,000 M(r) and isoelectric points 5.7 and 5.9, respectively, described previously by this laboratory, were found in the CSF of 27% of schizophrenics, 23% of the Alzheimer's disease patients, and 4% of the controls in the current study. The presence of proteins 127 and 128, as well as the increased concentrations of alpha-2 haptoglobin in the CSF of Alzheimer's disease and schizophrenic patients, may be useful as diagnostic biological markers. They may also indicate a common pathophysiology between these diseases.</p>","PeriodicalId":77007,"journal":{"name":"Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society","volume":"3 2","pages":"47-53"},"PeriodicalIF":0.0,"publicationDate":"1992-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"12457092","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Western blotting of transforming growth factor beta 2. Optimization of the electrophoretic transfer.","authors":"Y Jin,&nbsp;N Cerletti","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>The highest reported detection sensitivity of reduced monomeric transforming growth factor (TGF) beta 2 by Western blotting is 50 ng. The biologically active TGF-beta 2, which is the non-reduced dimeric protein, is even more difficult to detect by this technique. The low sensitivity is due to the poor electrophoretic transfer of the protein from the polyacrylamide gel to the blotting matrices under standard transfer conditions. By studying the effects of different blotting matrices, transfer cells, blotting buffers and their methanol content, current settings and the electrotransfer time, we have established optimal conditions for the blotting of this protein. The pH of the buffer and the blotting time are the most important factors which influence the electrotransfer yield. Optimal transfer of the TGF-beta 2 protein was achieved on PVDF membrane with semi-dry transfer for 4 h at 9 V, using 10 mM CAPS, pH 11.0, containing 5% methanol as transfer buffer. Combined with the use of a commercial antibody and an immunoblot assay kit, our optimised blotting method can detect 2 ng of the dimeric TGF-beta 2.</p>","PeriodicalId":77007,"journal":{"name":"Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society","volume":"3 2","pages":"85-90"},"PeriodicalIF":0.0,"publicationDate":"1992-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"12648148","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Electrokinetic and Kerr effect contributions to electric birefringence images of agarose electrophoresis gels.","authors":"M Lanan,&nbsp;M D Morris","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Multilinear regression analysis is used to test a model (Lanan et al., 1991) for electric birefringence imaging (EBI) of DNA fragment bands in agarose electrophoresis gels. EBI signals attributed to localized electro-kinetic gel distortion and to intrinsic DNA birefringence are studied by fitting ethidium bromide fluorescence profiles to EBI results. Fluorescence polarization imaging (FPI) is used to assess the influence of localized gel distortion on nucleic acid orientation across a fragment band. It is shown that DNA aligns parallel, on average, with an applied electric field independent of its location within a band. DNA orientation varies as the 1.2 power in electric field strength under the conditions tested.</p>","PeriodicalId":77007,"journal":{"name":"Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society","volume":"3 1","pages":"27-32"},"PeriodicalIF":0.0,"publicationDate":"1992-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"12766454","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Two-dimensional electrophoretic analyses of cod (Gadus morhua, L.) whole muscle proteins, water-soluble fraction and surimi. Effect of the addition of CaCl2 and MgCl2 during the washing procedure.","authors":"I Martinez,&nbsp;C Solberg,&nbsp;K Lauritzen,&nbsp;R Ofstad","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Samples from pre- and post-rigor cod mince, surimi (a concentrate of fish myofibrillar proteins obtained after washing and dewatering the fish mince) and water from the first wash in the surimi manufacture, processed with and without the addition of 7.5 mM CaCl2 and 15 mM MgCl2, were analyzed by two-dimensional electrophoresis. The results showed that the main myofibrillar proteins, including myosin, actin and tropomyosin, remained in the surimi. Several other proteins were selectively removed during the washing procedure. Some additional major spots were detected in the two-dimensional gels containing samples of the wash water and surimi processed with the addition of Ca2+ and Mg2+ salts. These spots were either absent or present in minor amounts in the samples of post-rigor cod mince, wash water and surimi processed without Ca2+ and Mg2+ salts and in all the pre-rigor samples. This induced us to suggest that the new additional spots may constitute fragments of proteins originated by increased proteolytic activity during the surimi manufacture upon the addition of the Ca2+ and Mg2+ salts. Two-dimensional electrophoresis has proved to be a valuable tool to quickly and easily assess the effect of different processing conditions on the protein content of the products.</p>","PeriodicalId":77007,"journal":{"name":"Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society","volume":"2 6","pages":"201-6"},"PeriodicalIF":0.0,"publicationDate":"1992-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"12735520","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Analysis of embryonic mouse development: construction of a high-resolution, two-dimensional gel protein database.","authors":"K E Latham,&nbsp;J I Garrels,&nbsp;C Chang,&nbsp;D Solter","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Numerous studies have revealed stage specific alterations in protein synthesis that occur in mouse embryos. A thorough analysis of these changes has been hampered by limitations in the ability to resolve individual proteins, the ability to accurately quantify and manage data from two-dimensional gel images, and by variation in the staging of the embryos used. To learn more of the changes in protein synthesis that occur during early development, we constructed a protein database for the mouse embryo using the QUEST system of high-resolution, two-dimensional gel electrophoresis and computerized gel image analysis (Garrels, 1989, J. Biol. Chem., 264:526). Synchronous cohorts of embryos were labeled at 3 h intervals throughout the entire preimplantation period from fertilization to blastocyst stage in order to characterize in detail the changes in protein synthesis pattern that occur during normal preimplantation development. Additional samples were prepared from early post-implantation embryos, isolated inner cell mass and trophoblast cells, and cultured embryonic stem cells. These provide the means for identifying cell and tissue specific proteins and for characterizing changes in protein synthesis that accompany early cellular differentiation in the embryo. We present here a description of the mouse embryo database and discuss its potential usefulness to the study of mammalian embryogenesis.</p>","PeriodicalId":77007,"journal":{"name":"Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society","volume":"2 6","pages":"163-70"},"PeriodicalIF":0.0,"publicationDate":"1992-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"12735571","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Two-dimensional polyacrylamide gel electrophoretic characterization of proteins from organs of C3H mice expressing the scurfy (sf) genetic mutation during early and late stages of disease progression.","authors":"J K Selkirk,&nbsp;M C Hite,&nbsp;V Godfrey,&nbsp;B A Merrick,&nbsp;C He,&nbsp;R A Griesemer,&nbsp;D R Daluge,&nbsp;B K Mansfield","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Scurfy (sf), is an X-linked recessive lethal mutation that occurs spontaneously in the C3H mouse. The disease is characterized by lymphoid and hematopoietic dysfunction. Affected males are of small stature and exhibit scaliness and crusting of the eyelids, ears, tail, and feet, marked splenomegaly, moderate hepatomegaly, enlarged lymph nodes, and atrophy of the thymus. The average lifespan of the affected hemizygous males (sf/y) is 24 +/- 0.7 days. Total cellular proteins were extracted from pooled samples of thymus and spleen obtained from combined litters of mice. Tissue-specific protein profiles characteristic of either sf mutant or normal mice were analyzed by two dimensional polyacrylamide gel electrophoresis (2DPAGE) at different stages of the phenotypic expression of the sf mutation, to identify changes in protein patterns that might be associated with the progression of the disease. The resultant gels were silver stained, digitized, and analyzed, by image analysis utilizing a pipelined image processor connected to a host computer. At 14 +/- 1 days of age, protein patterns from sf mutant and normal mice control organs showed considerable homogeneity, although there were proteins identified unique to the sf mutant and to the normal controls. At 20 +/- 1 days of age, the pattern differences between the sf mutant and normal control increased markedly. Differences were expressed as the percent of proteins that were unique to either the sf mutant or the normal control from the total number of each type. The percent of proteins that increased or decreased in the three organs utilized in this study ranged between 21%-39% at 14 days and were between 25%-54% at 20 days. Differences in protein expression between the normal and sf mutant as the disorder progressed for each of the three tissues examined. In addition, thymus protein profiles from 9 day old littermates that were phenotypically normal but genotypically unknown were evaluated to determine if marker proteins could be identified for the sf mutation. Limited protein changes were noted at relative molecular weights of 66, 60, 54, 39, 37, 33, 25, 23, 27, and 11 kDa. These data suggest that the sf mutation follows a trackable pattern of protein expression and repression different than the normal control C3H mouse. Several potential marker proteins associated with the sf mutation were identified in 9 day thymus prior to the phenotypic expression of the disease. These putative biomarkers may be useful for characterizing the sf mutation and the mutant may act a possible model the Wiskott-Aldrich syndrome (WAS).</p>","PeriodicalId":77007,"journal":{"name":"Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society","volume":"3 2","pages":"97-107"},"PeriodicalIF":0.0,"publicationDate":"1992-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"12648150","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Screening for antibodies against Aleutian disease virus (ADV) in mink. Elucidation of dubious results by additive counterimmunoelectrophoresis.","authors":"A Uttenthal","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>In order to distinguish true positive results in counterimmunoelectrophoresis from false positive ones an additive counterimmunoelectrophoresis was developed. The method was tested on selected mink serum samples as part of a routine testing for antibodies towards Aleutian disease virus on 3 million blood samples. The procedure of the method is, that a known positive serum sample is mixed with the patient serum to be tested. The result from a false positive sample will be one precipitin line towards virus and one nonspecific line. If the serum sample is a true positive one, the antibodies originating from the patient serum will be added to the antibodies in the standard positive serum giving only one precipitin line. The system is further extended by testing the serum samples towards an antigen preparation containing all the cellular components but free from virus.</p>","PeriodicalId":77007,"journal":{"name":"Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society","volume":"3 2","pages":"83-4"},"PeriodicalIF":0.0,"publicationDate":"1992-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"12508886","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Two-dimensional electrophoresis of urine specimens from patients with renal disease.","authors":"R P Tracy,&nbsp;D S Young,&nbsp;H D Hill,&nbsp;G W Cutsforth,&nbsp;D M Wilson","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>In an effort to develop non-invasive markers capable of characterizing renal disease with greater sensitivity and specificity than those currently available, urinary proteins were studied by high-resolution two-dimensional electrophoresis in order to catalog those proteins which appeared to be most affected by a variety of renal diseases. Urine specimens were prepared by high-pressure liquid chromatography and subjected to two-dimensional gel electrophoresis. Proteins were visualized with silver stain. Gels from 17 patients were analyzed in detail and compared to standard maps of human urinary proteins and plasma. The two-dimensional gels displayed approximately 100 times the number of proteins demonstrated by high resolution agarose electrophoresis. We have identified 34 proteins whose urinary concentrations were most affected by renal disease. Twenty seven were of plasma origin, based upon comigration with plasma samples (17 of unknown identity and function). Seven proteins were observed in normal urine but not present in plasma. These proteins, which may represent kidney tissue proteins, were not apparent in the urine of patients with proteinuria, most likely due to dilutional effects.</p>","PeriodicalId":77007,"journal":{"name":"Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society","volume":"3 2","pages":"55-65"},"PeriodicalIF":0.0,"publicationDate":"1992-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"12646807","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rapid and automated characterisation of seed genotype using Micrograd electrophoresis and pattern-matching software.","authors":"C W Wrigley,&nbsp;I L Batey,&nbsp;F Bekes,&nbsp;P J Gore,&nbsp;J Margolis","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>New precast microgels are described for use in quickly identifying seed of cereal varieties by determining protein composition within an hour. For example, gliadin proteins are extracted from crushed wheat grain, wheatmeal or flour with ethylene glycol (centrifugation not necessary) and 5 microliters extract is applied to a Micrograd gel (3-15% gel gradient) for ten minutes' electrophoresis at 300 volts in sodium lactate buffer (pH 3.1). Alternatively, precast gels are available for SDS gel electrophoresis for examining a different aspect of grain composition as a means of identification. To further expedite identification, software packages have been developed to match the protein pattern for an unknown sample against those of authentic samples, thus to provide quick and definite identity, based on electrophoretic banding, densitometer scan, HPLC profile, multiple antibody reaction or RFLP pattern (PatMatch program). Furthermore, the program WhatWheat offers advice on the best combination of methods to use for a specific task of identification.</p>","PeriodicalId":77007,"journal":{"name":"Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society","volume":"3 2","pages":"69-72"},"PeriodicalIF":0.0,"publicationDate":"1992-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"12648146","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Quantitation of pyrimidine dimers in DNA from UVB-irradiated alfalfa (Medicago sativa L.) seedlings.","authors":"F E Quaite,&nbsp;B M Sutherland,&nbsp;J C Sutherland","doi":"","DOIUrl":"","url":null,"abstract":"<p><p>Depletion of stratospheric ozone will increase the solar ultraviolet radiation in the range from 290-320 nm (UVB) that reaches the surface of the earth, placing an increased UV burden on exposed organisms. One consequence of increased UVB may be decreased productivity of crop plants. A principal lesion caused by UV in DNA is the cyclobutyl pyrimidine dimer. We have adapted a method for measuring these dimers in nanogram quantities of non-radioactive DNA for use in UV-irradiated plants. We find that biologically relevant doses of broad band UVB radiation induce easily detectable frequencies of pyrimidine dimers in the DNA of irradiated alfalfa sprout leaves and that the dose response for dimer formation is linear up to doses of at least 690 J m-2. We also find easily measurable frequencies of dimers in the leaves of seedlings grown in glass filtered sunlight but not exposed to additional UVB, suggesting that significant numbers of dimers are formed in plants exposed to normal sunlight.</p>","PeriodicalId":77007,"journal":{"name":"Applied and theoretical electrophoresis : the official journal of the International Electrophoresis Society","volume":"2 6","pages":"171-5"},"PeriodicalIF":0.0,"publicationDate":"1992-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"12735572","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}