Relative Mobility Values Research Articles

Biochemical Evaluation of Cotton Genotypes using Soluble Protein, Esterase (EST), Peroxidase (POX) And Polyphenol Oxidase (PPO) and their Role in Plant Disease Resistance

Isozyme analysis is a powerful biochemical technique that has numerous applications in plant biology. It has long been used by geneticists to study the population genetics. The isozyme esterase, peroxidase and polyphenol oxidase were standardized for upland cotton (Gossypium hirsutum L.) germplasm lines collected from all over the country. The knowledge of nature and magnitude of genetic diversity present in the germplasm is most important pre-requisite for the success of any breeding program. The thirty-four cotton germplasm lines were screened for prime three isozymes based on quantification assay and qualitative PAGE profiling. Among the material, the genotype AKH – 24 (190.60 mg ml-1), AKH – 053 (189.42 mg ml-1) and VIKAS (184.53 mg ml-1) recorded high protein content, whereas the enzymatic activities of esterase, peroxidase and polyphenol oxidase exhibited remarkable differences along with the protein content. The genotype LRA–5166 exhibited high esterase (462.68 mM mg protein-1 min-1) and peroxidase activity (250.97mM mg protein-1 min-1), while AKH – 24 recorded the maximum polyphenol oxidase activity (131.45 mM mg protein-1min-1). The banding pattern of biochemical markers revealed that the maximum number of bands were recorded in esterase analysis (fifteen) followed by protein (twelve) whereas, only five bands each were detected in peroxidase and polyphenol oxidase analysis indicating limited polymorphism. The Relative Mobility (Rm) values were ranged from 0.083 to 0.883 (protein), 0.100 to 0.971 (esterase), 0.033 to 0.283 (peroxidase) and 0.048 to 0.206 (polyphenol oxidase). The present study demonstrated that cotton genotypes could be differentiated by their quantity and quality through electrophoretic banding profiles. These results could be of practical value for cultivar identification, purity testing along with associated prediction of pest and disease resistance. However, the major constraint is that these biochemical markers do not able to reproduce the similar kind of variation pattern, but can provide strong distinguishing polymorphism each time.

Similarity Coefficient Analysis using UPGMA based on Polymorphism of Isozyme and DNA Markers in Cotton

Abstract: Cotton refers to those species of genus Gossypium that bears the spinnable seed coat fibres. The proteins and enzymes are the primary product of the genes and hence are most suitable for determination of genetic purity and polymorphism. The biochemical and molecular markers may complement the efforts to reduce the burden to a greater extent, as these are estimates devoid of environmental factor which a cause of error. Especially in the selection of parents based on biochemical and molecular polymorphism, there is marked improvement in the screens for the quantitative traits and understanding their architecture. The banding pattern of biochemical markers revealed that the maximum number of bands were recorded in esterase analysis (fifteen) followed by protein analysis (twelve) whereas, only five bands each were detected in peroxidase and polyphenol oxidase analysis indicating limited polymorphism. The Relative Mobility (Rm) values were ranged from 0.083 to 0.883 (protein), 0.100 to 0.971 (esterase), 0.033 to 0.283 (peroxidase) and 0.048 to 0.206 (polyphenol oxidase).

HEAT STRESS-INDUCED ALTERATIONS IN ANTIOXIDATIVE ENZYMES OF SOME PLANTS OF CUCURBITACEA FAMILY

The effects of high temperatures on melon cvs. Miranda and Poli, watermelon cv. Crimson Tide and zucchini cv. Asma leaves. The leaves obtained from plants were subjected to 35, 40, 45, 50, 55 and 60°C temperatures with gradual increments every 30-minutes. Samples, obtained at each treatment, were analyzed for ascorbic acid content, NADP(H) oxidase, catalase, gluthatione reductase, peroxidases activities and isoperoxidase patterns. The ascorbic acid content slightly increased parallel to temperature increment in zucchini but did not change in watermelon and in both melon cultivars. Melon cv. Poli exhibited comparatively less oxidative damage than cv. Miranda with a lower NAD(P)H oxidase activity. Heat stress induced NAD(P)H activity in watermelon and zucchini comparing to control plants. Results revealed that antioxidative enzyme activities were increased generally up to 50°C then decreased gradually in melon cultivars. Besides cv. Poli generally had higher enzyme activities than cv. Miranda. The activity of catalaes become prominent in watermelon while the activity of ascorbate peroxidase become prominent in zucchini. Acidic isoperoxidase bands with different relative mobility values were found in all species. Besides, basic isoperoxidase band could not be determined in both melon cultivars and watermelon while a basic isoperoxidase band was found in zucchini.

Relative Mobility (Rf) Analysis of Albumin Isolates from Snakehead Fish (Ophiocephalus striatus) Extracted at Different Temperatures and Times

Snakehead fish was a fish that has a high albumin content. Snakehead fish was used in the health sector known as medicinal freshwater fish, to accelerate the process of healing wounds after surgery and childbirth. The purpose of this study was to analyze the relative mobility (Rf) of the snakehead fish (Ophiocephalus striatus) albumin isolates by steaming at different temperatures and times. The research method for analyze the relative mobility (Rf) of the snakehead fish (Ophiocephalus striatus) albumin isolates by using gel filtration column chromatography, was detected by the SDS-PAGE electrophoresis method. Electrophoresis was carried out at a constant voltage of 125 v/ slab. Gel staining was carried out with Commasie Brilliant Blue R-250 in methanol-acetic acid-water. A standard protein mixture consisting of: myosin (200.0 kDa), β-galactosidase (116.3 kDa), phosphorylase B (97.4 kDa), bovine serum albumin (66, 2 kDa), ovalbumin (45.0 kDa), carbonic anhydrose (31.0 kDa), trypsin inhibitor (21.5 kDa), lysozyme (14.4 kDa) and aprotinim (6.5 kDa). The data analysis of this research was descriptive analysis to see the photos of the electrophoresis results. Meanwhile, for the analysis, the measurement of albumin isolation by gel filtration Sephadex G-75 was carried out with a divided plot design. The results showed that SDS-PAGE electrophoresis with the most complex amount of protein was albumin isolate, the effect of the steaming temperature was 40 oC for 30 minutes, located in 5 ml of the 1st fraction, 5 ml of the 2nd fraction and 5 ml of the 3rd fraction, with values of relative mobility (Rf) 0.0833 to 0.6944.

EgyGene GelAnalyzer4: a powerful image analysis software for one-dimensional gel electrophoresis

BackgroundGel image analysis is a cornerstone in electrophoresis-based experiments. Nature of the experiment determines the target of the analysis. Implemented features within different software for gel images and their outputs are variable. EgyGene GelAnalyzer4 software aimed to make a powerful, easy, and almost all-in-one software for gel image analysis. ResultsEgyGene GelAnalyzer4 is a software for analyzing the gel images. The program consists of six main parts: gel image enhancement, image analysis, data merging, molecular markers detection, phylogenetic tree prediction, and population parameters estimation. Firstly, the gel image enhancement part enhances the gel image colors and clarity. Then, the image is analyzed in the second part using both automatic and manual detection of lanes and bands. The third part, data merging, is for merging several analyses for different samples/gels together to produce one table comprises the entire experiment. The fourth section, markers detection, collects several pre-analyzed data for the same samples to detect their molecular markers based on some user-defined criteria. The fifth part uses the outputs of the previous parts to draw the phylogenetic tree between samples. The program constructs a phylogenetic tree using unweighted pair group method with arithmetic mean (UPGMA) method. Fourteen different equations are used to calculate the similarity percentages: Anderberg, Czekanowski, Dice, Faith, Gower and Legendre, Jaccard, Nei and Li, Rogers and Tanimoto, Russel and Rao, simple matching, Sokal and Michener, Sokal and Sneath 1, Sokal and Sneath 2, and Sorensen. The last section calculates the population parameters for the pre-analyzed images before. EgyGene GelAnalyzer4 is designed and programed using Microsoft Visual Studio 2012 Express version, based on the Dot Net FrameWork 4.5 edition, whereas its icons are created using the freeware Greenfish Icon Editor Pro 3.2.5.1. ConclusionsEgyGene GelAnalyzer4 is a simple, fast, and user-friendly software with six novel features: changing the order of lanes within the gel image, splitting the detecting lane into two lanes, merging of several detected lanes into just one, splitting a single band into two bands, merging two or more bands to be a single band, and comparing data of several analyzed images with a big number of samples and merging it together in a single table and detection of molecular markers between samples. In addition to the basic features and outputs of other image analysis software such as calculation of relative mobility values, molecular weights/sizes estimation, DNA/RNA/protein quantity within bands, table of present/absent of bands, polymorphism percentage, similarity percentages and phylogeny, population parameters, enhancing the image colors and clarity, and merging data for several gels. In addition, writing names of samples and the detected markers on the image. EgyGene GelAnalyzer4 provides many several features, which may be not available together in any other alternative.

ВЛИЯНИЕ НИЗКОМОЛЕКУЛЯРНЫХ ЭКЗОМЕТАБОЛИТОВ ФИТОПАТОГЕННЫХ И САПРОТРОФНЫХ МИКРОМИЦЕТОВ НА РОСТ, ФОРМИРОВАНИЕ БИОПЛЕНКИ И АНТАГОНИСТИЧЕСКУЮ АКТИВНОСТЬ PGPR ШТАММОВ BACILLUS И PAENIBACILLUS

By using of ultrafiltration procedure water-soluble peptide metabolites were isolated from the culture filtrates of several fungal strains including Bipolaris sorokiniana IB G-12, Fusarium oxysporum VKM F-137, Rhizoctonia solani VKM F-895 and Trichoderma harzianum IB G-58. Fractions of low molecular weight metabolites (Mw≥1 kDa) from all fungal strains were characterized with similar values of relative mobility (Rf ~ 0.37-0.39). Only extracellular metabolites of F. oxysporum and B. sorokiniana demonstrated bactericidal activity in agar-diffusion tests. Under tests of submerged liquid cultures, fungal metabolites selectively inhibited the growth of PGPR bacteria; the strain of Paenibacillus ehimensis IB-739 was most sensitive to their action. At the same time, fungal metabolites stimulated the growth of the B. subtilis IB-54 and nonspecific increase of its fungicidal activity, while enhancement in antifungal effect of the P. ehimensis IB-739 was species-specific. Most fungal metabolites suppressed under concentrations ≥ 0.25-0.5 mg/ml the development of fouling biofilms only in Bacillus strains that were characterized with high biofilm-forming ability in vitro. The synthesis of chitinolytic enzymes by the P ehimensis IB-739 strain was induced in presence of aqueous extracts from the some of micromycetes’ mycelia but not their extracellular metabolites.

Molecular Characterization of Trichoderma harzianum isolates from different Agroclimatic Zones of Karnataka.

Trichoderma species are the fungi that are present in nearly all soils and other diverse habitats. Trichoderma are common and are relatively easy to isolate and culture. In addition, isolates grow quickly on many substrates, produce metabolites with demonstrable antibiotic activity, and may be mycoparasitic against a wide range of pathogens (Grondona et al., 1997). Among Trichoderma genera Trichoderma harzianum is both plant growth promoter as well as antagonist against many pathogenic fungi. An effort was made to isolate ten isolates of Trichoderma harzianum from soil samples collected from different agroclimatic zones of Karnataka. A total of ten Trichoderma isolates representing each agroclimatic zone was isolated using Potato dextrose agar medium and purified. The fungal isolates were subjected to morphological tests and characterized as listed in The Manual of Soil Fungi (Gilman, 1961). Colony morphology and microscopic observation confirmed that all isolates belonging to the Trichoderma harzianum. Molecular diversity of ten isolates of Trichoderma harzianum was characterized by using SDSPAGE and RAPD markers. The data on the protein banding patterns of Trichoderma harzianum isolates were presented based on relative mobility value (Rm), squared euclidean distances and intensity of the bands. In RAPD a total of 125 bands were scored out of which 100 bands were found to be polymorphic (80%). Statistical analysis of both SDS-PAGE and RAPD data enabled the classification of 10 T.harzianum isolates in to two major groups. Thus SDS-PAGE and RAPD banding pattern of these isolates could easily distinguish the isolates of different zones.

Elution power of a solvent as a criterion of relative lipid polarity

New parameters are proposed that allow reliable calculation of fixed hydrophilicity values for different classes of lipids over the widest possible range, based on the elution power of solvents and using two compounds at the boundaries of the range as standards. The values of relative hydrophilicity are calculated from the values of relative chromatographic mobility of these types of compounds. It is established that the levels of hydrophilicity of different classes of lipids relative to the selected hexadecane–glycerol pair do not depend on the composition of the different mobile phases used in either planar or column types of liquid chromatography for the separation of complex lipid mixtures.

Electrophoretic mobility as a tool to separate immune adjuvant saponins from Quillaja saponaria Molina

Quillaja saponins are used as adjuvants in animal vaccines but their application in human vaccination is still under investigation. Isolation and characterization of adjuvant saponins is very tedious. Furthermore, standardization of Quillaja saponins is critical pertaining to its application in humans. In this study, a convenient method based on agarose gel electrophoresis was developed for the separation of Quillaja saponins. Six different commercial Quillaja saponins were segregated by size/charge into numerous fractions. Each of the fractions was characterized by ESI-TOF-MS spectroscopy and thin layer chromatography. Real-time impedance-based monitoring and red blood cell lysis assay were used to evaluate cytotoxicity and hemolytic activities respectively. Two specific regions in the agarose gel (delimited by specific relative electrophoretic mobility values) were identified and characterized by exclusive migration of acylated saponins known to possess immune adjuvant properties (0.18–0.58), and cytotoxic and hemolytic saponins (0.18–0.94). In vivo experiments in mice with the isolated fractions for evaluation of adjuvant activity also correlated with the relative electrophoretic mobility. In addition to the separation of specific Quillaja saponins with adjuvant effects as a pre-purification step to HPLC, agarose gel electrophoresis stands out as a new method for rapid screening, separation and quality control of saponins.

Community structure, diversity, and species dominance of bacteria, fungi, and nematodes from naturally and conventionally farmed soil: a case study on Japanese apple orchards

Although there has been much research on soil microbial communities in organic farming, little research has been reported on those in natural farming (no fertilizer use). Soil chemical properties and microbial and nematode communities in naturally (Orchard-N) and conventionally (Orchard-C) farmed apple orchards were compared over 3 years as a case study. The levels of nitrate nitrogen and available phosphate were significantly lower in Orchard-N than in Orchard-C. Bacterial evenness and nematode evenness and richness using denaturing gradient gel electrophoresis (DGGE) were higher in Orchard-N than in Orchard-C. All DGGE bands were identified by relative mobility values, termed relative front (Rf) values, and the Rf value of each band was classified into three dominant groups based on intensity: high, middle, and low intensities (HD, MD, and LD, respectively). Principal component analysis (PCA) on a correlation matrix showed that Orchard-N was characterized as MD/LD, corresponding to species of Alphaproteobacteria (bacteria), Actinobacteria (bacteria), Basidiomycota (fungi), Chaetomium (fungi), and Enoplea (nematodes). In contrast, Orchard-C was characterized by HD, corresponding to species of Gammaproteobacteria (bacteria), Fusarium (fungi), and Pratylenchus (nematodes), and MD/LD, similarly, Capnodiales (fungi) and Chloroflexi (bacteria). PCA on a variance–covariance matrix showed that annual changes in fungal and nematode community structures were larger in Orchard-C than in Orchard-N due to the large effect of HD. Quantitative polymerase chain reaction revealed that Orchard-N had higher bacterial and lower fungal abundance than Orchard-C. Based on these results, the relationship between the organism communities and the soil ecosystem in both orchards is discussed.

Biomolecular characterization of guar (Cyamopsis tetragonoloba) genotypes along with wild species, C. serrata and C. senegalensis

Accessibility to a wide inventory of the genetic diversity is desirable for genetic improvement of a crop. Rare availability of wild species which acts as reservoir for a number of important traits makes their biomolecular characterization highly significant. Cyamopsis tetragonoloba (L.) Taub. genotypes were subjected to molecular characterization by employing random amplified polymorphic DNA (54 genotypes) and seed storage protein profile (53 genotypes) along with two wild species, C. serrata Schinz. and C. senegalensis Guill. & Perr. The aim of this study was to analyse the extent of genetic variability, similarity and species identification. A total of 271 bands were amplified using 25 RAPD primers with an average of 10.84 bands per primer, whereas electrophoretic profile of seed storage proteins produced a total of 37 bands having relative mobility values ranging from 0.13 to 1.00, out of which 15 bands were polymorphic. A few DNA and protein bands specific to wild species were identified which showed that these are genetically distinct from C. tetragonoloba genotypes. Among 25 primers employed for the study, 9 produced a total of 12 unique alleles; however, some genotype/species-specific bands were also obtained in protein profiling. Jaccard’s coefficient-based genetic similarity indices ranged from 0.31 to 0.86 with an average of 0.58, indicating the presence of high levels of DNA polymorphism. But for seed storage proteins, similarity indices ranged from 0.70 to 1.00, indicating less protein variability. The dendrogram constructed by the UPGMA mode revealed genetic relationships among different C. tetragonoloba genotypes and among species. This study provides valuable information regarding characterization, identification, management, parental selection and seed purity that can be used in a synergistic way for augmenting the guar improvement programme in India.

ESTERASE ISOZYME PATTERNS DURING DEVELOPMENT OF BACTROCERA PAPAYA

One of the major horticultural crop pests is the Asian papaya fruit fly, Bactrocera papaya Drew & Hancock (Diptera: Tephritidae), in the tropical and subtropical regions of the world, which causes severe damage to the fleshy fruits and vegetables. Polyacrylamide gels were used for the first time to study the esterase isozyme patterns during different life stages and also to observe the enzyme patterns in the progeny (F1 generation) of B. papaya, which were maintained generation by generation under the laboratory condition. Four esterase isozymes, EST-1, EST-2, EST-3 and EST-4, were detected and their relative mobility values were 0.40, 0.26, 0.20 and 0.15, respectively. There was EST-1(0.40) with high mobility and EST-4(0.15) showed lowest mobility, closed to the cathode. These esterase isozymes were designated from the anodal end of the gel, i.e., the highest to lowest relative mobility. EST-1(0.40), EST-20.26 and EST-3(0.20) bands were only present in larval stage. EST-4(0.15) band was observed in all developmental stages except eggs. The thickness and the staining degree of bands varied in different developmental stages. In larvae, EST-1(0.40) band was very thin, EST-2(0.26) band was very thick and highly stained and EST-3(0.20) band was thin. The band of EST-4(0.15) was thick in larval and pupal stages, and comparatively very thick and highly stained in adults of both males and females. These variations of esterase isozymes indicated that it had stage-specific patterns in this pest species. In the progeny (F1 generation) of B. papaya, especially larvae and adults showed the similar esterase isozyme band patterns, comparing with their parents. The findings of the present research could be used for distinguishing this pest from other Bactrocera species as well as helpful to develop an environment friendly control method for this pest.

Alogliptin, but not voglibose, ameliorates dyslipidemia and LDL particle size in patients with impaired glucose tolerance or Type 2 diabetes

Aim: To compare the potential amelioration of dyslipidemia and LDL particle size by alogliptin and voglibose in patients with impaired glucose tolerance (IGT) or Type 2 diabetes mellitus (T2DM). Patients & methods: Patients (n = 37; 25 patients with IGT and 12 patients with newly diagnosed T2DM) were randomly assigned to treatment with either alogliptin (25 mg daily for 12 weeks) or voglibose (0.6 mg daily for 12 weeks). A 75-g oral glucose tolerance test and laboratory measurements were performed at baseline and after treatment. LDL particle size was evaluated by electrophoresis of lipoproteins. Results: Following treatment, alogliptin, but not voglibose, decreased triglycerides (p < 0.01) and increased HDL cholesterol (p < 0.05), and significantly decreased the relative mobility value (p < 0.05). Conclusion: Alogliptin ameliorates dyslipidemia and increases LDL particle size in patients with IGT or T2DM.

Geochemistry of Cd in groundwater of Winder, Balochistan and suspected health problems

Sedimentary rocks of Jurassic age (Ferozabad group) and igneous rocks (Bela Ophiolite) of Cretaceous age are widely exposed in the east of Winder Town, Balochistan. Representative 48 water and soil samples were collected and analyzed for Cd and other elements. The present study showed that water samples were contaminated by the Cd ion and most of the samples have higher concentration than prescribed WHO standards (3 μg/l) for drinking purpose. The concentration of Cd ions ranged between 1 and 30 with mean values of 10 μg/l and was found high in the vicinity of outcrops. The alkaline pH (av. 7.42) was mainly responsible for elevated Cd content in the water, having near equal molar proportions of Ca and HCO3 ions. Computed relative mobility values appeared as 0.23, indicating poor leaching from rocks and metal load in the groundwater is also measured low. The estimated values of mobility reflect fairly high mobility of Cd in the study area, decreases with distance from outcrops. The health hazard indices of drinking water was high (av. 1.33) with respect to safe average daily intake of Cd. The consumption of high Cd-bearing water may induce many disorders in the human health.

Biotic Stress Induced Biochemical and Isozyme Variations in Ginger and Tomato by &amp;lt;i&amp;gt;Ralstonia solanacearum&amp;lt;/i&amp;gt;

This unique study evaluates the effects of Ralstonia solanacearum (Rs) induced biotic stress in two cultivars, Zingiber officinale (ginger) and Lycopersicon esculentum (tomato). They were grown in pots and hydroponic systems with controls; to induce biotic stress, about 8 × 104 colony forming units of Rs suspension was injected into the healthy test plants. Upon induction of Rs stress, highly significant (p 0.01) biochemical changes (%) were noticed in respect to controls: carbohydrate content was generally high in both plants; while they showed decreased starch and protein contents; phenolics showed a swing of decrease or increase between pot and hydroponic systems; and all plants in general showed higher (3-6 fold) proline content upon induction of biotic stress. Regarding oxidative stress isozymes (OSE), superoxide dismutase (EC 1.15.1.1) isozymes were normally 3, but treated hydroponics had 4 with comparable relative mobility values; peroxidase (EC 1.11.1.7) isozymes were generally 2, except for treated hydroponic tomato. Briefly, Rs induced biotic stress caused wilt symptoms in ginger, but did not affect tomato though its biochemical and OSE patterns especially in those grown as hydroponics were elicited to significantly higher levels.

Leaf Protein Electrophoresis and Taxonomy of Species of Jatropha L. (Euphorbiaceae)

The systematic relationship existing among members of the all important genus Jatropha was studied using leaf protein electrophoresis. The aim was to identify possible taxonomic importance of the protein profile in the estimation and elucidation of the taxonomic affinity of the six species of Jatropha (Jatropha curcas Linn., J. podagrica Hook., J. gossypifolia Linn., J. mutifida Linn., J. tanjorensis Ellis &amp; Saroja and J. integerrima Linn.) found in Nigeria. The species were screened for total protein banding patterns using gel electrophoresis. Young leaves (0.8 g) of the plants were washed with distilled water and macerated with sterile mortar and pestle in 0.8% Phosphate Buffer-Saline (PBS) containing 0.4 M NaCl at pH 8.0. Results reveal that protein banding pattern was taxon specific. Generic band occurs at 8.3. The highest number of interspecific bands (4) exists between J. podagrica and J. multifida. Variations exist not only in the number of bands but also in the intensity of the bands. Sokal and Sneath coefficient of similarity ranges between 11.1-44.4 %. Single linkage Cluster Analysis (SLCA) of the relative mobility values of the protein in the taxa shows partial agreement with current sub generic and sectional delimitation of the species based on morphology and anatomy of the species.

Abstract 4782: Two-dimensional gel analysis of protein expression profile in patients with invasive squamous cell carcinoma of the uterine cervix for identification of differentially expressed proteins in relation to response to radiation therapy

Abstract Background: Cancer of uterine cervix is the second most common cancer and an important cause of cancer related mortality among women worldwide. These patients are treated by chemoradiation, response to which is a major factor affecting their outcome. Objective: To identify differentially expressed proteins in the tissue biopsies of patients with invasive cervical cancer by two-dimensional gel electrophoresis [2-DE] and correlate it with their clinical outcome. Methods: After an informed consent, 10 patients of invasive squamous cervical cancer, stage IIB, between 40-50 years treated with chemoradiation as per standard protocol were recruited. Based on the 24 month-outcome, there were two groups (N=5): i) Evidence of disease (ED) and ii) No evidence of disease (NED). Pooled protein from the tissue biopsies were subjected to 2-DE using pH 3-10 IPG strips of 7cm length, silver stained and expression profile compared using Image Master 7.0 software (GE Healthsciences). The differentially expressed protein spots were identified and isoelectric point (pI) &amp; Molecular Weight (MW) was determined from the plot of relative electrophoretic mobility (Rf) values of the marker against their respective molecular weight and the plot of pI marker against their respective distance from the anode respectively. The TagIdent software was used for their identification. Results: Out of a total of 327 spots identified, 89 spots showed differential expression, of which 40 were highly significant (ANOVA p &amp;lt;0.03) between ED vs NED groups. Conclusion: 2-DE has provided preliminaryinformation regarding differentially expressed proteins which may help identify biomarkers for prediction of outcome in patients with invasive cervical cancer which needs further confirmation. Differentially expressed proteins in NED vs ED groups in invasive cervical cancer Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4782. doi:1538-7445.AM2012-4782

High-level resistance to gentamicin: genetic transfer between Enterococcus faecalis isolated from food of animal origin and human microbiota

To investigate the in vivo gene transfer of high-level gentamicin resistance (HLRG) from Enterococcus faecalis isolated from the food of animal origin to a human isolate, using a mouse model of intestinally colonized human microbiota. In vitro study: The presence of plasmids involved in HLRG coding was investigated. After the conjugation experiment, the recipient strain, Ent. faecalis JH2-SS, acquired a plasmid responsible for HLRG [minimal inhibitory concentration (MIC) >800 μg ml(-1) ], in a similar position to the donor cells. In vivo study: Seven BALB/c mice were dosed with ceftriaxone (400 mg kg(-1) ) and then inoculated with a dilution of 1/100 of human faeces (HFc). After 72 h, Ent. faecalis JH2-SS (recipient) was inoculated and then, after a further 72 h, the animals were given Ent. faecalis CS19, isolated from the food of animal origin, involved in HLRG (donor). The presence of transconjugant strains in HFc was subsequently recorded on a daily basis until the end of the experiment. The clonal relationship between Ent. faecalis and Escherichia coli in faeces was assessed by RAPD-PCR. Both the in vitro and in vivo studies showed that the receptor strain acquired a plasmid responsible for HLRG (MICs >800 μg ml(-1) ), which migrated with a similar relative mobility value. Transconjugant strains were detected from 24 h after the donor strain inoculation and persisted until the end of the experiment. The in vivo gene transfer of HLRG from Ent. faecalis strains, isolated from the food of animal origin, to human microbiota has been demonstrated in a mouse model. The complexity found on the therapeutic responses of invasive infectious diseases caused by Ent. faecalis facilitates the assessment of food of animal origin as a resistant pathogen reservoir. In addition, this study may contribute to the understanding of antimicrobials' resistance gene transfer between Ent. faecalis strains from food and human GI tract.

BIOCHEMICAL MARKERS: A USEFUL TOOL FOR ASSESSING GENETIC DIVERSITY IN JACKFRUIT (ARTOCARPUS HETEROPHYLLUS LAM.)

Using polyacrylamide gel electrophoresis, peroxidase and esterase isozymes were investigated from leaf tissue samples of 44 selected (superior) jackfruit genotypes. These selected genotypes were collected following jackfruit descriptor 2000, after surveying 1500 trees in the districts of Nadia, 24 Parganas (N) and Coochbehar in West Bengal. The selected genotypes were analyzed for isozyme variation for peroxidase and esterase. Polymorphism was observed in both the enzyme systems studied, where several bands were found showing similarities and dissimilarities among the selected types. In the peroxidase assay, 7 loci were identified among the 44 genotypes. The relative mobility (R m ) values of the loci ranged between 0.29 and 0.50. Whereas in the esterase assay, 6 loci were identified and the relative mobility (R m ) values of the loci ranged between 0.22 and 0.65. Moreover, it was observed that genotypes collected from nearby locations did not show similar banding patterns, indicating that forces other than geographical origin, such as exchange of genetic stocks, genetic drift, spontaneous variation, natural and artificial selection, cross pollination and seedling origin of the plant are responsible for the genetic diversity.

Effects of High Intensity Exercise on Isoelectric Profiles and SDS-PAGE Mobility of Erythropoietin

Exercise induced proteinuria is a common phenomenon in high performance sports. Based on the appearance of so called "effort urines" in routine doping analysis the purpose of this study was to investigate the influence of exercise induced proteinuria on IEF profiles and SDS-PAGE relative mobility values (rMVs) of endogenous human erythropoietin (EPO). Twenty healthy subjects performed cycle-ergometer exercise until exhaustion. VO (2)max, blood lactate, urinary proteins and urinary creatinine were analysed to evaluate the exercise performance and proteinuria. IEF and SDS-PAGE analyses were performed to test for differences in electrophoretic behaviour of the endogenous EPO before and after exercise. All subjects showed increased levels of protein/creatinine ratio after performance (8.8+/-5.2-26.1+/-14.4). IEF analysis demonstrated an elevation of the relative amount of basic band areas (13.9+/-11.3-36.4+/-12.6). Using SDS-PAGE analysis we observed a decrease in rMVs after exercise and no shift in direction of the recombinant human EPO (rhEPO) region (0.543+/-0.013-0.535+/-0.012). Following identification criteria of the World Anti Doping Agency (WADA) all samples were negative. The implementation of the SDS-PAGE method represents a good solution to distinguish between results influenced by so called effort urines and results of rhEPO abuse. Thus this method can be used to confirm adverse analytical findings.