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Electrophoretic Analysis (electrophoretic + analysis)
Selected AbstractsTwo-Dimensional Electrophoretic Analysis of Soluble Leaf Proteins of a Salt-sensitive (Triticum aestivum) and a Salt-tolerant (T. durum) Cultivar in Response to NaCl StressJOURNAL OF INTEGRATIVE PLANT BIOLOGY, Issue 7 2007Mustafa YILDIZ Abstract In this research, 3-day-old etiolated wheat seedlings of Triticum aestivum L. cv. Ceyhan-99 (salt-sensitive) and T. durum Desf. cv. F,rat-93 (salt-tolerant) were grown in control and salt (150 mmol/L NaCl) treatments at a 15/25 °C temperature regime in the light for 12 days. Soluble proteins extracted from the first leaf tissues of two cultivars were analyzed by two-dimensional (2-D) electrophoresis in order to detect NaCl-induced changes. The soluble leaf protein profiles of cultivars were observed to be similar. However, quantitative differences in 74 proteins were detected in the salt treatment group, compared to the control. Among the 74 protein spots, 14 were common for two cultivars. As a result of NaCl treatment, two low-molecular-weight (LMW) proteins (28.9 and 30.0 kDa) and one intermediate-molecular-weight (IMW) protein (44.3 kDa) in cv. Ceyhan-99 and six LMW proteins (18.6, 19.4, 25.7, 25.9, 26 and 27.6 kDa) in cv. F,rat-93 were newly synthesized. The newly synthesized proteins were specific to each cultivar. In the F,rat-93 cultivar, four proteins with LMW (24.8,27.9 kDa) were completely lost in NaCl treatment. Moreover, these four protein spots were not observed in both protein profiles of cv. Ceyhan-99. Most of these proteins were in acidic character (pl <6.0,6.9) and low molecular weight (<31.6 kDa). It is suggested that the newly synthesized or completely lost LMW proteins may be important for cultivars differing in sensitivity towards NaCl. [source] Gonadoinhibitory effects of Neb-colloostatin and Neb-TMOF on ovarian development in the mealworm, Tenebrio molitor L.ARCHIVES OF INSECT BIOCHEMISTRY AND PHYSIOLOGY (ELECTRONIC), Issue 3 2007O. Wasielewski Abstract The gonadostatic action of the peptides Neb-colloostatin (SIVPLGLPVPIGPIVVGPR) and Neb-TMOF (NPTNLH) from Neobellieria bullata was studied in female mealworm Tenebrio molitor. Both peptides potently inhibit ovarian development and terminal oocyte maturation of mated females during their first reproductive cycle. Injection of 4 ,g of Neb-colloostatin or Neb-TMOFNeb-TMOF reduced, at day 4 of the cycle, the size of the terminal oocytes to about half or one third of the normal size in saline-injected controls. In addition, follicular patency was arrested. The injections of Neb-colloostatin and Neb-TMOF also caused a delay to the first ovulation and oviposition as well as a reduction of the number of eggs by about 50% in the first 3 days of the oviposition period. At 4 days after adult emergence, none of the peptides had caused significant changes in protein concentration or composition of the haemolymph. However, both peptides reduced total protein content in ovaries and induced qualitative changes in ovarian protein patterns. Electrophoretic analyses indicated that Neb-colloostatin and Neb-TMOF caused a loss of two proteins (150, 180 kDa) and a drastic reduction of 4 others (39, 43, 47, 130 kDa), which are the most abundant ones in ovaries of control females. On the other hand, they increased the concentration of 2 other polypeptides (65, 70 kDa), which normally occur in insignificant quantities in ovaries. Our results indicate that both peptides have a very similar mode of action despite the differences in their amino acid sequence. They seem to interfere with vitellogenin production by the fat body as well as with vitellogen uptake by the oocytes through modification of patency. Arch. Insect Biochem. Physiol. 64:131,141, 2007. © 2007 Wiley-Liss, Inc. [source] Electrophoretic analysis of sequence variability in three mitochondrial DNA regions for ascaridoid parasites of human and animal health significanceELECTROPHORESIS, Issue 13 2008Ming-Wei Li Abstract Sequence variability in three mitochondrial DNA (mtDNA) regions, namely cytochrome c oxidase subunit 1 (cox1), NADH dehydrogenase subunits 1 and 4 (nad1 and nad4), among and within Toxocara canis, T. cati, T. malaysiensis, T. vitulorum and Toxascaris leonina from different geographical origins was examined by a mutation-scanning approach. A portion of the cox1 gene (pcox1), a portion of the nad1 and nad4 genes (pnad1 and pnad4) were amplified separately from individual ascaridoid nematodes by polymerase chain reaction and the amplicons analyzed by single-strand conformation polymorphism (SSCP). Representative samples displaying sequence variation in SSCP profiles were subjected to sequencing in order to define genetic markers for their specific identification and differentiation. While the intra-specific sequence variations within each of the five ascaridoid species were 0.2,3.7% for pcox1, 0,2.8% for pnad1 and 0,2.3% for pnad4, the inter-specific sequence differences were significantly higher, being 7.9,12.9% for pcox1, 10.7,21.1% for pnad1 and 12.9,21.7% for pnad4, respectively. Phylogenetic analyses based on the combined sequences of pcox1, pnad1 and pnad4 revealed that the recently described species T. malaysiensis was more closely related to T. cati than to T. canis. These findings provided mtDNA evidence for the validity of T. malaysiensis and also demonstrated clearly the usefulness and attributes of the mutation-scanning sequencing approach for studying the population genetic structures of these and other nematodes of socio-economic importance. [source] Inter-ocean dispersal is an important mechanism in the zoogeography of hakes (Pisces: Merluccius spp.)JOURNAL OF BIOGEOGRAPHY, Issue 6 2001W. Stewart Grant Aim To present new genetic data and to review available published genetic data that bear on the phylogeny of hakes in the genus Merluccius. To construct a zoogeographical model from a summary phylogenetic tree with dated nodes. To search for an explanation of antitropical distributions in hakes. To assess peripheral isolate, centrifugal and vicariance models of speciation in view of the molecular phylogeny and zoogeography of hakes. Locations Northern and southern Atlantic Ocean, eastern Pacific Ocean, South Pacific Ocean. Methods Electrophoretic analysis of 20 allozyme loci in 10 species of hakes. Phylogenetic tree construction with parsimony and bootstrap methods. Reanalysis of previous genetic data. Analysis of zoogeographical patterns with geographical distributions of molecular genetic markers. Results Phylogenetic analyses of new and previous allozyme data and previous mitochondrial DNA data indicate a deep genetic partition between Old- and New-World hakes with genetic distances corresponding to 10,15 Myr of separation. This time marks a widening rift between Europe and North America and a rapid drop in ocean temperatures that subdivided an ancestral population of North Atlantic hake. Two Old-World clades spanning the equator include pairs of sister taxa separated by tropical waters. Divergence times between these pairs of sister-taxa variously date to the early Pliocene and late Pleistocene. Amongst New-World hakes, pairs of sister taxa are separated by equatorial waters, by the Southern Ocean, and by the Panama Isthmus. These genetic separations reflect isolation by the rise of the Isthmus 3,4 Ma and by Pliocene and Pleistocene dispersals. Pairs of species occurring in sympatry or parapatry in six regions do not reflect sister-species relationships, but appear to reflect allopatric divergence and back dispersals of descendent species. Some geographically isolated regional populations originating within the last few hundreds of thousands of years merit subspecies designations. Conclusions Vicariance from tectonic movement of continental plates or ridge formation cannot account for the disjunct distributions of most hake sister taxa. Molecular genetic divergences place the origin of most hake species diversity in the last 2,3 Myr, a period of negligible tectonic activity. Distributions of many hake species appear to have resulted from dispersals and back dispersals across both warm equatorial waters and cool waters in the Southern Ocean, driven by oscillations in climate and ocean temperatures. Genetic and ecological divergence prevents hybridization and competitive exclusion between sympatric species pairs in six regions. Sister-taxa relationships and estimates of divergence are consistent with the modified peripheral isolate model of speciation in which vicariances, range expansions and contractions, dispersals and founder events lead to isolated populations that subsequently diverge to form new species. [source] Extraction of native collagen from limed bovine split wastes through improved pretreatment methodsJOURNAL OF CHEMICAL TECHNOLOGY & BIOTECHNOLOGY, Issue 7 2008Dong Li Abstract BACKGROUND: The large amount of limed bovine split wastes discharged by the leather industry has raised concerns regarding their environmental effect. The objective of this work was to perform pilot plant trials to extract high-value native collagen from these wastes through improved pretreatment methods. RESULTS: EDTA- and HCl-pretreatment gave similar removal percentages of inorganic substances. Owing to the open structure of fibers, the collagen yield of HCl-pretreated splits (HPS) (41.31%) was higher than that of EDTA-pretreated splits (EPS) (10.42%). Furthermore, HCl-pretreated split collagen (HPC) had a more acidic isoelectric point, lower content of primary amino groups, larger Z-average particle size and higher relative viscosity than EDTA-pretreated split collagen (EPC). Electrophoretic analysis and circular dichroism spectra revealed the maintenance of polypeptide and triple helix conformation, respectively. In addition, the transition temperatures of EPC (34.7 °C) and HPC (34.6 °C) detected by differential scanning calorimetry (DSC) were close to that of commercial collagen from calfskin (CCC) (35.7 °C). CONCLUSION: A process of native collagen extraction from limed bovine split wastes was proposed. While both EPC and HPC represented similar physicochemical properties to those of CCC, the collagen yield of HPS was much higher than that of EPS. Copyright © 2008 Society of Chemical Industry [source] Electrophoretic analysis of urinary proteins in diabetic adolescentsJOURNAL OF CLINICAL LABORATORY ANALYSIS, Issue 4 2001George Koliakos Abstract Pathological changes in the urine sodium dodecyl sulphate gel electrophoresis (SDS PAGE) patterns often precede the occurrence of any sign of renal involvement in diabetes. However, data concerning the most frequent SDS PAGE pattern of the urine in early stages of type I diabetes mellitus are controversial. In the present study an SDS PAGE technique has been used that provides an adequate sensitivity for the detection of the abnormal pattern. Urinary proteins have been analyzed by SDS PAGE in twenty two diabetic adolescents and twenty four age matched controls. Albumin concentration, and N acetyl-beta-D-glucosaminidase (NAG) activity were also measured in the same samples. There was no significant difference in urine albumin concentration and NAG activity between diabetic children and controls. However twelve patients showed an electrophoretic pattern characteristic for glomerulopathy, two had a pattern indicating tubular dysfunction and another two patients had a mixed pattern. Among the twenty four controls only three showed abnormal electrophoretic patterns. The results support the view that early stages of diabetic nephropathy may involve both glomerular and tubular dysfunction. However the exact clinical and prognostic significance of the information provided by SDS PAGE analysis remains to be elucidated. J. Clin. Lab. Anal. 15:178,183, 2001. © 2001 Wiley-Liss, Inc. [source] Pollination ecology, genetic diversity and selection on nectar spur length in Platanthera lacera (Orchidaceae)PLANT SPECIES BIOLOGY, Issue 3 2005KAREN J. LITTLE Abstract Platanthera lacera (Orchidaceae) is a moth-pollinated, loess prairie orchid producing a raceme of one to many whitish-green flowers. Field studies on a western Illinois population found the crepuscular visiting noctuid moth, Anagrapha falcifera (Noctuidae), to be the most frequent pollinator with occasional visits from Allagrapha aerea (Noctuidae). Visitation rates, assessed by removal of at least one pollinium, were relatively high (84.9%) and fruit production on experimentally outcrossed flowers (94.4%) was higher than open-pollinated plants (71.4%). Experimental pollination showed P. lacera to be highly self-compatible (94.1%) with a low level of autogamy (8.2%). Measurements taken from 598 spurs on 44 plants indicated that nectar spur length varied significantly among plants (10.9,17.1 mm, mean 14.3 mm), but was not under selective pressure from visitation by An. falcifera (mean proboscis length 11.1 mm). The absence of selective pressure on nectar spur length is likely to be explained by occasional pollinating visits from Al. aerea (proboscis length 18 mm) and a limited amount of autogamy. Electrophoretic analysis of 12 enzymes revealed seven polymorphic loci. Mean levels of heterozygosity were He = 0.3384, Ho = 0.3229 and F = 0.0458, indicating that P. lacera is primarily an outcrossing species dependent on noctuid moth pollination. [source] Potato yellow vein virus: its host range, distribution in South America and identification as a crinivirus transmitted by Trialeurodes vaporariorumANNALS OF APPLIED BIOLOGY, Issue 1 2000L F SALAZAR Summary Sporadic outbreaks of potato yellow vein disease (PYVD) were first observed in the early 1940's by potato growers in Antioquia, Colombia. Long known to be transmitted by the greenhouse whitefly (Trialeurodes vaporariorum), the precise identity of its causal agent (presumably viral in nature) has remained obscure. Here, we present evidence that a closterovirus with a bipartite genome, potato yellow vein virus (PYVV), is associated with PYVD. Electrophoretic analysis revealed that diseased tissue contains 4,5 disease-specific dsRNAs ranging in size from c. 9 000,1 800 bp. RT-PCR reactions containing pairs of degenerate primers directed against conserved motifs in the closterovirus heat-shock protein homologue produced products of the expected sizes. Comparison of the corresponding amino acid sequences revealed striking similarities between PYVV and two bipartite, whitefly-transmitted criniviruses, Cucurbit yellow stunting disorder and Tomato chlorosis viruses. Epidemiological surveys carried out in Rionegro, Colombia identified Polygonum mepalense, Polygonum spp., Rumex obtusifolium, Tagetes spp., and Catharanthus roseus as potential viral reservoirs. PYVV is transmitted through tubers, and visual symptoms alone cannot be used to determine infection status. A sensitive hybridisation-based assay for PYVV has been developed for use in seed certification programmes. [source] Ultrastructure, Eneystment and Cyst Wall Composition of the Resting Cyst of the Peritrich Ciliate Opisthonecta henneguyiTHE JOURNAL OF EUKARYOTIC MICROBIOLOGY, Issue 1 2003PURIFICACIÓN CALVO ABSTRACT. The cyst wall of Opisthonecta henneguyi has been studied ultrastructurally and cytochemically by light and electron microscopy, as well as by chemical and electrophoretic analyses, to examine the structure of the cyst wall and its composition. The cyst wall consists of four morphologically distinct layers. The ectocyst is a thin dense layer. The mesocyst is the thickest layer and is composed of a compact material. The endocyst is a thin layer like the ectocyst, but less dense. The granular layer varies in thickness and is composed of a granular material. In the resting cyst, kinetosomes of both oral apparatus and trochal band as well as the myoneme system are maintained, and only cilia are resorbed. The sugars present in the cyst wall are predominantly N-acetylglucosamine (90%) and glucose (10%). The niesocyst is composed of chitin, and the endocyst includes glycoproteins and acid mucopolysaccharides. During secretion of the cyst wall, the endocyst and granular layer are secreted from precursors synthesized "de novo". No cytoplasmic precursors of ectocyst and mesocyst have been detected. [source] Lectin-based electrophoretic analysis of the expression of the 35,kDa inter-,-trypsin inhibitor heavy chain H4 fragment in sera of patients with five different malignanciesELECTROPHORESIS, Issue 12 2008Emida Mohamed Abstract A 35,kDa glycoprotein whose abundance was previously demonstrated to be enhanced in sera of patients with endometrial adenocarcinoma (n,=,12), was isolated from pooled sera of three of the cancer patients using champedak galactose-binding lectin affinity chromatography in the present study. Subjecting it to 2-DE and MS/MS, the glycoprotein was identified as the O -glycosylated fragment of inter-,-trypsin inhibitor heavy chain H4 (ITIH4). When compared to control sera (n,=,17), expression of the 35,kDa ITIH4 cleavage fragment was demonstrated to be significantly enhanced in sera of patients with breast carcinoma (n,=,10), epithelial ovarian carcinoma (n,=,10), and germ cell ovarian carcinoma (n,=,10) but not in patients with nasopharyngeal carcinoma (n,=,13) and osteosarcoma (n,=,7). The lectin-based electrophoretic bioanalytical method adopted in the present study may be used to assess the physiological relevance of ITIH4 fragmentation and its correlation with different malignancies, their stages and progression. [source] The potential of inductively coupled plasma-mass spectrometric detection for capillary electrophoretic analysis of pesticidesELECTROPHORESIS, Issue 17 2005Rodolfo G. Wuilloud See original http://dx.doi.org/10.1002/elps.200410098 [source] Integrated on-chip derivatization and electrophoresis for the rapid analysis of biogenic aminesELECTROPHORESIS, Issue 14 2004Nigel P. Beard Abstract We demonstrate the monolithic integration of a chemical reactor with a capillary electrophoresis device for the rapid and sensitive analysis of biogenic amines. Fluorescein isothiocyanate (FITC) is widely employed for the analysis of amino-group containing analytes. However, the slow reaction kinetics hinders the use of this dye for on-chip labeling applications. Other alternatives are available such as o -phthaldehyde (OPA), however, the inferior photophysical properties and the UV ,max present difficulties when using common excitation sources leading to a disparity in sensitivity. Consequently, we present for the first time the use of dichlorotriazine fluorescein (DTAF) as a superior in situ derivatizing agent for biogenic amines in microfluidic devices. The developed microdevice employs both hydrodynamic and electroosmotic flow, facilitating the creation of a polymeric microchip to perform both precolumn derivatization and electrophoretic analysis. The favorable photophysical properties of the DTAF and its fast reaction kinetics provide detection limits down to 1 nM and total analysis times (including on-chip mixing and reaction) of <60 s. The detection limits are two orders of magnitude lower than current limits obtained with both FITC and OPA. The optimized microdevice is also employed to probe biogenic amines in real samples. [source] Transcription factor binding study by capillary zone electrophoretic mobility shift assayELECTROPHORESIS, Issue 1-2 2003Zsolt Ronai Abstract Regulation of gene expression through interaction of proteins with specific DNA sequences is a central issue in functional genomics. Capillary electrophoretic mobility shift assay is an efficient novel method for the investigation of sequence specific protein-DNA interactions, allowing rapid and sensitive quantification of the complex formation. In this paper, we present a pilot study on capillary zone electrophoretic mobility shift assay (CZEMSA) to investigate the interaction between the transcription factors of HeLa nuclear extract and Sp1-specific fluorescein-labeled oligonucleotide, using the unlabeled probe as competitor. The mobility shift assay was accomplished by CZE in coated capillaries without polymeric buffer additives. Specificity of the DNA protein complex formation was verified by competition experiments, as well as by supershift assay with an anti-Sp1 antibody. The applied electric field strength did not affect the stability of DNA-protein complex during the electrophoretic analysis, allowing rapid identification and quantification of the protein DNA interaction. A practical application to study the interaction between Oryza sativa MADS-box transcription factor 4 (OsMADS4) and its consensus sequence is also reported. [source] Apoptotic effect of cyanobacterial extract on rat hepatocytes and human lymphocytesENVIRONMENTAL TOXICOLOGY, Issue 3 2001Joanna Mankiewicz Abstract Toxic cyanobacterial blooms are an increasing problem in Poland. The production of cyanobacterial toxins and their presence in drinking and recreational waters represent a growing danger to human and animal health. This is connected with the increase of cyanobacterial biomass caused by excessive eutrophication of the water ecosystem. There is evidence that cyanobacterial hepatotoxins can act as a potent promoter of primary liver cancer. The apoptotic effect of microcystins in Polish cyanobacterial bloom samples on rat hepatocytes and human lymphocytes was observed using light and fluorescence microscopy, flow cytometry, and electrophoretic analysis. The incubation time needed to observe the first morphological apoptotic changes in hepatocytes was approximately 30 min; however, the characteristic biochemical changes in DNA were not observed even after 120 min. In lymphocyte cultures the morphological changes characteristic for apoptosis were observed after 24 h of incubation and a 48-h incubation was found to be optimal for analysis of internucleosomal DNA fragmentation, which is one of the main biochemical hallmarks of programmed cell death. These cells are an easily isolated and inexpensive material for medical diagnostics. Therefore the apoptotic changes, together with the clastogenic effect seen in lymphocyte cultures, are proposed as a future analytical method for these toxins. © 2001 John Wiley & Sons, Inc. Environ Toxicol 16: 225,233, 2001 [source] Bacterial leaf blight of strawberry (Fragaria (x) ananassa) caused by a pathovar of Xanthomonas arboricola, not similar to Xanthomonas fragariae Kennedy & King.PLANT PATHOLOGY, Issue 6 2001Description of the causal organism as Xanthomonas arboricola pv. fragariae (pv. nov., comb. nov.) A new bacterial disease of strawberry is described. This disease, called bacterial leaf blight of strawberry, is characterized by dry, brown necrotic leaf spots and large brown V-shaped lesions along the leaf margin, midrib and major veins. Symptoms are different from angular leaf spot of strawberry caused by the bacterium Xanthomonas fragariae. Strains of the bacterial leaf blight pathogen were characterized in a polyphasic approach by biochemical tests, fatty acid analysis, protein electrophoresis, serology, PCR, pigment analysis, ice-nucleation activity, AFLP analysis, DNA:DNA hybridization, pathogenicity and host range tests, and compared with a number of reference strains of X. fragariae and other Xanthomonas species. Bacterial leaf blight strains formed a homogeneous group in all tests, completely different from X. fragariae. They were the only strains causing leaf blight of strawberry upon artificial inoculation into strawberry. Fatty acid and protein electrophoretic analysis showed that the strains belong to the phenon X. campestris (sensu latu, including pathovars now classified as belonging to X. arboricola). AFLP analysis and DNA:DNA hybridization further clarified their taxonomic position as belonging to X. arboricola. The name X. arboricola pv. fragariae is proposed for the bacterium causing leaf blight of strawberry with strain PD2780 (LMG 19145) as pathovar type strain. Criteria for routine identification are given and the taxonomic status is discussed. [source] Genetic relationship between Litopenaeus setiferus (L.) and L. schmitti (Burkenroad) determined by using 16S mitochondrial sequences and enzymatic analysisAQUACULTURE RESEARCH, Issue 12 2003L Arena Abstract Genetic differentiation and variability data of two populations of two species of shrimp (Litopenaeus setiferus (L.) and L. schmitti (Burkenroad)) have been obtained by electrophoretic analysis and by analysis of 16S mitochondrial DNA. Using eight polymorphic enzymes, the genetic distance (GD) between the two species was 0.165. The GD between L. setiferus populations was 0.0057 and between L. schmitti populations it was 0.0034. The greatest differentiation was found within, rather than between, populations, although the differentiation value between Mexican and Cuban populations varied in accordance with the geographic distance and ecological characteristic of each. We found a high similarity between these two species with a bimodal distribution of the loci with respect to genetic identity. The homology percentages for gene 16S fragments were compared with those from six different shrimp species (L. vannamei, L. stylirostris, Farfantepenaeus notialis, Metapeneopsis lamellata) and Artemia salina. Ninety-seven percent of identity was found by analysis of a 409 bp of 16S mitochondrial DNA. With these values a phylogenetic tree was made using parsimony criteria. The GDs obtained with this method confirm the classification proposed by Pérez-Farfante & Kensley (1997). [source] | ||||||||||||||||