Search results for: nephrotoxicity markers
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 755

Search results for: nephrotoxicity markers

695 Pioglitazone Ameliorates Methotrexate-Induced Renal Endothelial Dysfunction via Amending Detrimental Changes in Antioxidant Profile, Systemic Cytokines and Apoptotic Factors

Authors: Sahar M. El-Gowilly, Mai M. Helmy, Hanan M. El-Gowelli

Abstract:

Methotrexate (MTX) is widely used in treatment of cancers and autoimmune diseases. However, nephrotoxicity is one of the most important side effects of MTX. The peroxisome proliferator-activated receptor gamma agonist, pioglitazone (PIO), is known to exert anti-inflammatory and reno-protective effects in various kidney injuries. The purpose of this study was to investigate the potential involvement of endothelial damage in MTX-induced renal injury and to elaborate the possible protective effect of PIO against MTX-induced nephropathy. Compared with saline-treated rats, treatment with MTX (7 mg/kg for 3 day) caused significant elevations in serum levels of urea and creatinine, increased renal nitrate/nitrite level and impaired renovascular responsiveness of isolated perfused kidney to endothelium-dependent vasodilations induced by acetylcholine (0.01-2.43 nmol) and isoprenaline (1µmol). These effects were abolished by concurrent treatment with PIO (2.5 mg/kg, for 5 days starting two days before MTX). Alternatively, MTX treatment did not affect endothelium-independent renovascular relaxation induced by sodium nitroprusside (1-30 μmole). The possibility that alterations in renal antioxidants, circulating cytokine and apoptotic factor (Fas) levels contributed to MTX-PIO interaction was assessed. PIO treatment abrogated renal oxidative stress (decreased reduced glutathione and catalase activity and increased malondialdehyde), elevated serum cytokine (interleukin-6, interleukin-10, tumor necrosis factor-alpha and transforming growth factor-beta1) and Fas induced by MTX. Histologically, MTX caused defused tubular cells swelling and vacuolization associated with endothelial damage in renal arterioles. These effects disappeared upon co-treated with PIO. Collectively, PIO abolished MTX-induced endothelium dysfunction and nephrotoxicity via ameliorating oxidative stress and rectifying cytokines and Fas abnormalities caused by MTX.

Keywords: methotrexate, pioglitazone, endothelium, kidney

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694 Pioglitazone Ameliorates Methotrexate-Induced Renal Endothelial Dysfunction via Amending Detrimental Changes in Antioxidant Profile, Systemic Cytokines and Fas Production

Authors: Sahar M. El-Gowilly, Mai M. Helmy, Hanan M. El-Gowelli

Abstract:

Methotrexate (MTX) is widely used in treatment of cancers and autoimmune diseases. However, nephrotoxicity is one of its most important side effects. The peroxisome proliferator-activated receptor gamma agonist, pioglitazone, is known to exert antiinflammatory and reno-protective effects in various kidney injuries. The purpose of this study was to investigate the potential involvement of endothelial damage in MTX-induced renal injury and to elaborate the possible protective effect of pioglitazone against MTX-induced endothelial impairment. Compared with saline-treated rats, treatment with MTX (7 mg/kg for 3 day) caused significant elevations in serum levels of urea and creatinine, increased renal nitrate/nitrite level and impaired renovascular responsiveness of isolated perfused kidney to endothelium-dependent vasodilations induced by acetylcholine (0.01-2.43 nmol) and isoprenaline (1µmol). These effects were abolished by concurrent treatment with pioglitazone (2.5 mg/kg, for 5 days starting two days before MTX). Alternatively, MTX treatment did not affect endothelium-independent renovascular relaxation induced by sodium nitroprusside (0.001-10 μmole). The possibility that alterations in renal antioxidants, circulating cytokine and apoptotic factor (Fas) levels contributed to MTX-pioglitazone interaction was assessed. Pioglitazone treatment abrogated renal oxidative stress (decreased reduced glutathione and catalase activity and increased malondialdehyde), elevated serum cytokine (interleukin-6, interleukin-10, tumor necrosis factor-alpha and transforming growth factor-beta1) and Fas induced by MTX. Histologically, MTX caused defused tubular cells swelling and vacuolization associated with endothelial damage in renal arterioles. These effects disappeared upon co-treated with pioglitazone. Collectively, pioglitazone abolished MTX-induced endothelium dysfunction and nephrotoxicity via ameliorating oxidative stress and rectifying cytokines and Fas abnormalities caused by MTX.

Keywords: methotrexate, pioglitazone, endothelium, kidney

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693 Determination of Genetic Markers, Microsatellites Type, Liked to Milk Production Traits in Goats

Authors: Mohamed Fawzy Elzarei, Yousef Mohammed Al-Dakheel, Ali Mohamed Alseaf

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Modern molecular techniques, like single marker analysis for linked traits to these markers, can provide us with rapid and accurate genetic results. In the last two decades of the last century, the applications of molecular techniques were reached a faraway point in cattle, sheep, and pig. In goats, especially in our region, the application of molecular techniques is still far from other species. As reported by many researchers, microsatellites marker is one of the suitable markers for lie studies. The single marker linked to traits of interest is one technique allowed us to early select animals without the necessity for mapping the entire genome. Simplicity, applicability, and low cost of this technique gave this technique a wide range of applications in many areas of genetics and molecular biology. Also, this technique provides a useful approach for evaluating genetic differentiation, particularly in populations that are poorly known genetically. The expected breeding value (EBV) and yield deviation (YD) are considered as the most parameters used for studying the linkage between quantitative characteristics and molecular markers, since these values are raw data corrected for the non-genetic factors. A total of 17 microsatellites markers (from chromosomes 6, 14, 18, 20 and 23) were used in this study to search for areas that could be responsible for genetic variability for some milk traits and search of chromosomal regions that explain part of the phenotypic variance. Results of single-marker analyses were used to identify the linkage between microsatellite markers and variation in EBVs of these traits, Milk yield, Protein percentage, Fat percentage, Litter size and weight at birth, and litter size and weight at weaning. The estimates of the parameters from forward and backward solutions using stepwise regression procedure on milk yield trait, only two markers, OARCP9 and AGLA29, showed a highly significant effect (p≤0.01) in backward and forward solutions. The forward solution for different equations conducted that R2 of these equations were highly depending on only two partials regressions coefficient (βi,) for these markers. For the milk protein trait, four marker showed significant effect BMS2361, CSSM66 (p≤0.01), BMS2626, and OARCP9 (p≤0.05). By the other way, four markers (MCM147, BM1225, INRA006, andINRA133) showed highly significant effect (p≤0.01) in both backward and forward solutions in association with milk fat trait. For both litter size at birth and at weaning traits, only one marker (BM143(p≤0.01) and RJH1 (p≤0.05), respectively) showed a significant effect in backward and forward solutions. The estimates of the parameters from forward and backward solution using stepwise regression procedure on litter weight at birth (LWB) trait only one marker (MCM147) showed highly significant effect (p≤0.01) and two marker (ILSTS011, CSSM66) showed a significant effect (p≤0.05) in backward and forward solutions.

Keywords: microsatellites marker, estimated breeding value, stepwise regression, milk traits

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692 Studying Language of Immediacy and Language of Distance from a Corpus Linguistic Perspective: A Pilot Study of Evaluation Markers in French Television Weather Reports

Authors: Vince Liégeois

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Language of immediacy and distance: Within their discourse theory, Koch & Oesterreicher establish a distinction between a language of immediacy and a language of distance. The former refers to those discourses which are oriented more towards a spoken norm, whereas the latter entails discourses oriented towards a written norm, regardless of whether they are realised phonically or graphically. This means that an utterance can be realised phonically but oriented more towards the written language norm (e.g., a scientific presentation or eulogy) or realised graphically but oriented towards a spoken norm (e.g., a scribble or chat messages). Research desiderata: The methodological approach from Koch & Oesterreicher has often been criticised for not providing a corpus-linguistic methodology, which makes it difficult to work with quantitative data or address large text collections within this research paradigm. Consequently, the Koch & Oesterreicher approach has difficulties gaining ground in those research areas which rely more on corpus linguistic research models, like text linguistics and LSP-research. A combinatory approach: Accordingly, we want to establish a combinatory approach with corpus-based linguistic methodology. To this end, we propose to (i) include data about the context of an utterance (e.g., monologicity/dialogicity, familiarity with the speaker) – which were called “conditions of communication” in the original work of Koch & Oesterreicher – and (ii) correlate the linguistic phenomenon at the centre of the inquiry (e.g., evaluation markers) to a group of linguistic phenomena deemed typical for either distance- or immediacy-language. Based on these two parameters, linguistic phenomena and texts could then be mapped on an immediacy-distance continuum. Pilot study: To illustrate the benefits of this approach, we will conduct a pilot study on evaluation phenomena in French television weather reports, a form of domain-sensitive discourse which has often been cited as an example of a “text genre”. Within this text genre, we will look at so-called “evaluation markers,” e.g., fixed strings like bad weather, stifling hot, and “no luck today!”. These evaluation markers help to communicate the coming weather situation towards the lay audience but have not yet been studied within the Koch & Oesterreicher research paradigm. Accordingly, we want to figure out whether said evaluation markers are more typical for those weather reports which tend more towards immediacy or those which tend more towards distance. To this aim, we collected a corpus with different kinds of television weather reports,e.g., as part of the news broadcast, including dialogue. The evaluation markers themselves will be studied according to the explained methodology, by correlating them to (i) metadata about the context and (ii) linguistic phenomena characterising immediacy-language: repetition, deixis (personal, spatial, and temporal), a freer choice of tense and right- /left-dislocation. Results: Our results indicate that evaluation markers are more dominantly present in those weather reports inclining towards immediacy-language. Based on the methodology established above, we have gained more insight into the working of evaluation markers in the domain-sensitive text genre of (television) weather reports. For future research, it will be interesting to determine whether said evaluation markers are also typical for immediacy-language-oriented in other domain-sensitive discourses.

Keywords: corpus-based linguistics, evaluation markers, language of immediacy and distance, weather reports

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691 Comparison of Developed Statokinesigram and Marker Data Signals by Model Approach

Authors: Boris Barbolyas, Kristina Buckova, Tomas Volensky, Cyril Belavy, Ladislav Dedik

Abstract:

Background: Based on statokinezigram, the human balance control is often studied. Approach to human postural reaction analysis is based on a combination of stabilometry output signal with retroreflective marker data signal processing, analysis, and understanding, in this study. The study shows another original application of Method of Developed Statokinesigram Trajectory (MDST), too. Methods: In this study, the participants maintained quiet bipedal standing for 10 s on stabilometry platform. Consequently, bilateral vibration stimuli to Achilles tendons in 20 s interval was applied. Vibration stimuli caused that human postural system took the new pseudo-steady state. Vibration frequencies were 20, 60 and 80 Hz. Participant's body segments - head, shoulders, hips, knees, ankles and little fingers were marked by 12 retroreflective markers. Markers positions were scanned by six cameras system BTS SMART DX. Registration of their postural reaction lasted 60 s. Sampling frequency was 100 Hz. For measured data processing were used Method of Developed Statokinesigram Trajectory. Regression analysis of developed statokinesigram trajectory (DST) data and retroreflective marker developed trajectory (DMT) data were used to find out which marker trajectories most correlate with stabilometry platform output signals. Scaling coefficients (λ) between DST and DMT by linear regression analysis were evaluated, too. Results: Scaling coefficients for marker trajectories were identified for all body segments. Head markers trajectories reached maximal value and ankle markers trajectories had a minimal value of scaling coefficient. Hips, knees and ankles markers were approximately symmetrical in the meaning of scaling coefficient. Notable differences of scaling coefficient were detected in head and shoulders markers trajectories which were not symmetrical. The model of postural system behavior was identified by MDST. Conclusion: Value of scaling factor identifies which body segment is predisposed to postural instability. Hypothetically, if statokinesigram represents overall human postural system response to vibration stimuli, then markers data represented particular postural responses. It can be assumed that cumulative sum of particular marker postural responses is equal to statokinesigram.

Keywords: center of pressure (CoP), method of developed statokinesigram trajectory (MDST), model of postural system behavior, retroreflective marker data

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690 Interactive Multiple Functions User Interface

Authors: Manjit Singh Sidhu, Waleed Maqableh, Jee Geak Ying

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Tangible user interfaces (TUI) that employ markers in the augmented reality (AR) environment has hampered the interactivity between the user and the software application. This is because the user lacks focus on visualizing the contents due to the interaction mechanisms whereby multiple markers may need to be used to perform a particular function. In this research, we have designed a novel TUI user interface where multiple functions could be triggered similar to a natural keyboard thus allowing user to focus more on its digital contents such as 2D/3D, text input, animation and sound. Test results of the user interface with potential users and HCI experts revealed that the multiple functions user interface was new, preferred and appreciated more as opposed to marker based user interface.

Keywords: multimedia, augmented reality, engineering, user interface, visualization

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689 Rebamipide Retards CCL4 Induced Hepatic Fibrosis: A Role of PGE2

Authors: Alaa E. El-sisi, Sherin Zakaria

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Rebamipide is an antiulcer drug with unique properties such as anti-inflammatory action. It induces endogenous prostaglandin e2 (PGE2). PGE2 is considered as a potent physiological suppressor of liver fibrosis. Aim of study: This study investigated the effect of rebamipide on hepatic fibrosis. Material and Method: Hepatic fibrosis was induced by intraperitoneal injections (IP) injection of CCl4 (0.45 mL/kg) in corn oil 1:5 twice a week for 4 weeks. Rats were divided into four groups as follow: Group 1 treated with CCL4 only, group 2 and 3 treated with CCL4 and rebamipide 60 mg/kg/day (group2) or 100 mg/kg/day (group3), and the fourth group was considered as control group and treated with vehicles. ALT, AST, and Bilirubin were assayed in serum. Antioxidant markers such as malondialdhyde (MDA) and superoxide dismutase (SOD) and fibrotic markers such as hyaluronic acid (HA) and procollagen-III (procol-III) were evaluated in liver tissues. IL-10 as well as PGE2 were also assayed in liver tissues. Pathologic changes in the liver were detected by hematoxylin and eosin staining. Collagen precipitation in liver tissues was visualized using masson trichrom stain. Results: Rebamipide inhibit CCL4 induced increase in ALT and AST significantly (p < 0.05). Rebamipide exerted an antioxidant effect as it inhibits CCL4 induced increased MDA level and decreased SOD activity. Fibrotic markers assay revealed that repamipide (60 or 100 mg/kg/day) decreased the level of procol-III and HA compared to CCl4 (p < 0.05). Oral administration of Rebamipide was associated with a significant increase (p < 0.05) of PGE2 and IL-10. Rebamipide especially at the dose of (100 mg/kg/day) restores liver histology structure and abolish collagen precipitation in liver tissues. Conclusion: Rebamipide retards hepatic fibrosis induced by CCL4 may be through the induction of PGE2 level.

Keywords: fibrotic markers, hepatic fibrosis, PGE2, rebamipide

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688 Microfluidic Lab on Chip Platform for the Detection of Arthritis Markers from Synovial Organ on Chip by Miniaturizing Enzyme-Linked ImmunoSorbent Assay Protocols

Authors: Laura Boschis, Elena D. Ozzello, Enzo Mastromatteo

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Point of care diagnostic finds growing interest in medicine and agri-food because of faster intervention and prevention. EliChip is a microfluidic platform to perform Point of Care immunoenzymatic assay based on ready-to-use kits and a portable instrument to manage fluidics and read reliable quantitative results. Thanks to miniaturization, analyses are faster and more sensible than conventional ELISA. EliChip is one of the crucial assets of the Europen-founded Flamingo project for in-line measuring inflammatory markers.

Keywords: lab on chip, point of care, immunoenzymatic analysis, synovial arthritis

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687 Preliminary Analysis of a Phylogeography Study of Dendropsophus minutus in the Guiana Shield

Authors: Mera-Martínez Daniela

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Dendropsophus minutus, is a species distributed in South America including the slopes of the Andes, the Amazon basin, forests of southeastern Brazil and in Guyana where tropical forests are characteristic. The relationship of amphibians found in this locality is evidenced by molecular markers, with the objective of analyzing if the geographic distance is influencing the structure of the populations of D. minutus in Guyana; we analyzed 65 sequences from the 3 localities of Guyana where haplotype networks, Mantel Test and phylogeny were realized to know the influence. It was evidenced that there is a haplotypic difference in the locality of Guyana compared to Suriname and French Guyana, but this does not have a correlation with the geographic distance, but this one can be influenced by the conditions of the places.

Keywords: phylogeography, Dendropsophus, geographic distance, molecular markers

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686 ISSR Based Molecular Phylogeny in Naturally Growing Suaeda Populations of Saudi Arabia

Authors: Mohammed Abdullah Basahi

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The objective of the present study was to identify the phylogenetic relationships and determine genetic diversity among Suaeda genotypes growing in Saudi Arabia and to find out whether these could be a potential source for genetic diversity. A set of nineteen genotypes was analyzed using twenty-four ISSR primers. Clear amplified polymorphic DNA products were obtained from the screening of twenty-four ISSR primers on nineteen genotypes that allowed selection of ten primers and the results were reproducible. Nineteen genotypes were revealed a unique profile with ten ISSR primers and thus it can be used for the DNA fingerprinting. Different primers produced a different level of polymorphism among the nineteen genotypes. The number of polymorphic bands per primer varied from 5 to 14 with an average of 8 bands per primer. The results revealed that the genotypes differed for ISSR markers. The genetic similarity based on Nei and Li’s ranged from 0.450 to 0.930. Cluster analysis was conducted based on ISSR data to group the Suaeda genotypes and to construct a dendrogram. Four groups can be distinguished by truncating the dendrogram at GS value of 0.54. ISSR markers showed high level of polymorphism among the genotypes examined. The present study indicates that ISSR markers could be successfully used in genetic characterization and diversity in Suaeda.

Keywords: suaeda, DNA fingerprinting, ISSR, Saudi Arabia

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685 Nephrotoxicity and Hepatotoxicity Induced by Chronic Aluminium Exposure in Rats: Impact of Nutrients Combination versus Social Isolation and Protein Malnutrition

Authors: Azza A. Ali, Doaa M. Abd El-Latif, Amany M. Gad, Yasser M. A. Elnahas, Karema Abu-Elfotuh

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Background: Exposure to Aluminium (Al) has been increased recently. It is found in food products, food additives, drinking water, cosmetics and medicines. Chronic consumption of Al causes oxidative stress and has been implicated in several chronic disorders. Liver is considered as the major site for detoxification while kidney is involved in the elimination of toxic substances and is a target organ of metal toxicity. Social isolation (SI) or protein malnutrition (PM) also causes oxidative stress and has negative impact on Al-induced nephrotoxicity as well as hepatotoxicity. Coenzyme Q10 (CoQ10) is a powerful intracellular antioxidant with mitochondrial membrane stabilizing ability while wheat grass is a natural product with antioxidant, anti-inflammatory and different protective activities, cocoa is also potent antioxidants and can protect against many diseases. They provide different degrees of protection from the impact of oxidative stress. Objective: To study the impact of social isolation together with Protein malnutrition on nephro- and hepato-toxicity induced by chronic Al exposure in rats as well as to investigate the postulated protection using a combination of Co Q10, wheat grass and cocoa. Methods: Eight groups of rats were used; four served as protected groups and four as un-protected. Each of them received daily for five weeks AlCl3 (70 mg/kg, IP) for Al-toxicity model groups except one group served as control. Al-toxicity model groups were divided to Al-toxicity alone, SI- associated PM (10% casein diet) and Al- associated SI&PM groups. Protection was induced by oral co-administration of CoQ10 (200mg/kg), wheat grass (100mg/kg) and cocoa powder (24mg/kg) combination together with Al. Biochemical changes in total bilirubin, lipids, cholesterol, triglycerides, glucose, proteins, creatinine and urea as well as alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), lactate deshydrogenase (LDH) were measured in serum of all groups. Specimens of kidney and liver were used for assessment of oxidative parameters (MDA, SOD, TAC, NO), inflammatory mediators (TNF-α, IL-6β, nuclear factor kappa B (NF-κB), Caspase-3) and DNA fragmentation in addition to evaluation of histopathological changes. Results: SI together with PM severely enhanced nephro- and hepato-toxicity induced by chronic Al exposure. Co Q10, wheat grass and cocoa combination showed clear protection against hazards of Al exposure either alone or when associated with SI&PM. Their protection were indicated by the significant decrease in Al-induced elevations in total bilirubin, lipids, cholesterol, triglycerides, glucose, creatinine and urea levels as well as ALT, AST, ALP, LDH. Liver and kidney of the treated groups also showed significant decrease in MDA, NO, TNF-α, IL-6β, NF-κB, caspase-3 and DNA fragmentation, together with significant increase in total proteins, SOD and TAC. Biochemical results were confirmed by the histopathological examinations. Conclusion: SI together with PM represents a risk factor in enhancing nephro- and hepato-toxicity induced by Al in rats. CoQ10, wheat grass and cocoa combination provide clear protection against nephro- and hepatotoxicity as well as the consequent degenerations induced by chronic Al-exposure even when associated with the risk of SI together with PM.

Keywords: aluminum, nephrotoxicity, hepatotoxicity, isolation and protein malnutrition, coenzyme Q10, wheatgrass, cocoa, nutrients combinations

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684 Endothelial Progenitor Cells Is a Determinant of Vascular Function and Atherosclerosis in Ankylosing Spondylitis

Authors: Ashit Syngle, Inderjit Verma, Pawan Krishan

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Objective: Endothelial progenitor cells (EPCs) have reparative potential in overcoming the endothelial dysfunction and reducing cardiovascular risk. EPC depletion has been demonstrated in the setting of established atherosclerotic diseases. With this background, we evaluated whether reduced EPCs population are associated with endothelial dysfunction, subclinical atherosclerosis and inflammatory markers in ankylosing spondylitis (AS) patients without any known traditional cardiovascular risk factor in AS patients. Methods: Levels of circulating EPCs (CD34+/CD133+), brachial artery flow-mediated dilatation, carotid intima-media thickness (CIMT) and inflammatory markers i.e erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), tissue necrosis factor (TNF)–α, interleukin (IL)-6, IL-1 were assessed in 30 AS patients (mean age33.41 ± 10.25; 11 female and 19 male) who fulfilled the modified New York diagnostic criteria with 25 healthy volunteers (mean age 29.36± 8.64; 9 female and 16 male) matched for age and sex. Results: EPCs (CD34+/CD133+) cells were significantly (0.020 ± 0.001% versus 0.040 ± 0.010%, p<0.001) reduced in patients with AS compared to healthy controls. Endothelial function (7.35 ± 2.54 versus 10.27 ±1.73, p=0.002), CIMT (0.63 ± 0.01 versus 0.35 ± 0.02, p < 0.001) and inflammatory markers were also significantly (p < 0.01) altered as compared to healthy controls. Specifically, CD34+CD133+cells were inversely multivariate correlated with CRP and TNF-α and endothelial dysfunction was positively correlated with reduced number of EPC. Conclusion: Depletion of EPCs population is an independent predictor of endothelial dysfunction and early atherosclerosis in AS patients and may provide additional information beyond conventional risk factors and inflammatory markers.

Keywords: endothelial progenitor cells, atherosclerosis, ankylosing spondylitis, cardiovascular

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683 Genetic Diversity of Sorghum bicolor (L.) Moench Genotypes as Revealed by Microsatellite Markers

Authors: Maletsema Alina Mofokeng, Hussein Shimelis, Mark Laing, Pangirayi Tongoona

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Sorghum is one of the most important cereal crops grown for food, feed and bioenergy. Knowledge of genetic diversity is important for conservation of genetic resources and improvement of crop plants through breeding. The objective of this study was to assess the level of genetic diversity among sorghum genotypes using microsatellite markers. A total of 103 accessions of sorghum genotypes obtained from the Department of Agriculture, Forestry and Fisheries, the African Centre for Crop Improvement and Agricultural Research Council-Grain Crops Institute collections in South Africa were estimated using 30 microsatellite markers. For all the loci analysed, 306 polymorphic alleles were detected with a mean value of 6.4 per locus. The polymorphic information content had an average value of 0.50 with heterozygosity mean value of 0.55 suggesting an important genetic diversity within the sorghum genotypes used. The unweighted pair group method with arithmetic mean clustering based on Euclidian coefficients revealed two major distinct groups without allocating genotypes based on the source of collection or origin. The genotypes 4154.1.1.1, 2055.1.1.1, 4441.1.1.1, 4442.1.1.1, 4722.1.1.1, and 4606.1.1.1 were the most diverse. The sorghum genotypes with high genetic diversity could serve as important sources of novel alleles for breeding and strategic genetic conservation.

Keywords: Genetic Diversity, Genotypes, Microsatellites, Sorghum

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682 Morphological and Molecular Characterization of Accessions of Black Fonio Millet (Digitaria Iburua Stapf) Grown in Selected Regions in Nigeria

Authors: Nwogiji Cletus Olando, Oselebe Happiness Ogba, Enoch Achigan-Dako

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Digitaria iburua, commonly known as black fonio, is a cereal crop native to Africa and extensively cultivated by smallholder farmers in Northern Benin, Togo, and Nigeria. This crop holds immense nutritional and socio-cultural value. Unfortunately, limited knowledge about its genetic diversity exists due to a lack of scientific attention. As a result, its potential for improvement in food and agriculture remains largely untapped. To address this gap, a study was conducted using 41 accessions of D. iburua stored in the genebank of the Laboratory of Genetics, Biotechnology, and Seed Science at Abomey-Calavi University, Benin. The study employed both morphological and simple sequence repeat (SSR) markers to evaluate the genetic variability of the accessions. Agro-morphological assessments were carried out during the 2020 cropping season, utilizing an alpha lattice design with three replications. The collected data encompassed qualitative and quantitative traits. Additionally, molecular variability was assessed using eleven SSR markers. The results revealed significant phenotypic variability among the evaluated accessions, leading to their classification into three main clusters. Furthermore, the eleven SSR markers identified a total of 50 alleles, averaging 4.55 alleles per locus. The primers exhibited an average polymorphic information content value of 0.43, with the DE-ARC019 primer displaying the highest value (0.59). These findings suggest a substantial degree of genetic heterogeneity within the evaluated accessions, and the SSR markers employed in the study proved highly effective in detecting and characterizing this genetic variability. In conclusion, this study highlights the presence of significant genetic diversity in black fonio and provides valuable insights for future efforts aimed at its genetic improvement and conservation.

Keywords: genetic diversity, digitaria iburua, genetic improvement, simple sequence repeat markers, Nigeria, conservation

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681 TiO₂ Nanotube Array Based Selective Vapor Sensors for Breath Analysis

Authors: Arnab Hazra

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Breath analysis is a quick, noninvasive and inexpensive technique for disease diagnosis can be used on people of all ages without any risk. Only a limited number of volatile organic compounds (VOCs) can be associated with the occurrence of specific diseases. These VOCs can be considered as disease markers or breath markers. Selective detection with specific concentration of breath marker in exhaled human breath is required to detect a particular disease. For example, acetone (C₃H₆O), ethanol (C₂H₅OH), ethane (C₂H₆) etc. are the breath markers and abnormal concentrations of these VOCs in exhaled human breath indicates the diseases like diabetes mellitus, renal failure, breast cancer respectively. Nanomaterial-based vapor sensors are inexpensive, small and potential candidate for the detection of breath markers. In practical measurement, selectivity is the most crucial issue where trace detection of breath marker is needed to identify accurately in the presence of several interfering vapors and gases. Current article concerns a novel technique for selective and lower ppb level detection of breath markers at very low temperature based on TiO₂ nanotube array based vapor sensor devices. Highly ordered and oriented TiO₂ nanotube array was synthesized by electrochemical anodization of high purity tatinium (Ti) foil. 0.5 wt% NH₄F, ethylene glycol and 10 vol% H₂O was used as the electrolyte and anodization was carried out for 90 min with 40 V DC potential. Au/TiO₂ Nanotube/Ti, sandwich type sensor device was fabricated for the selective detection of VOCs in low concentration range. Initially, sensor was characterized where resistive and capacitive change of the sensor was recorded within the valid concentration range for individual breath markers (or organic vapors). Sensor resistance was decreased and sensor capacitance was increased with the increase of vapor concentration. Now, the ratio of resistive slope (mR) and capacitive slope (mC) provided a concentration independent constant term (M) for a particular vapor. For the detection of unknown vapor, ratio of resistive change and capacitive change at any concentration was same to the previously calculated constant term (M). After successful identification of the target vapor, concentration was calculated from the straight line behavior of resistance as a function of concentration. Current technique is suitable for the detection of particular vapor from a mixture of other interfering vapors.

Keywords: breath marker, vapor sensors, selective detection, TiO₂ nanotube array

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680 Metadiscourse in Chinese and Thai Request Emails: Analysis and Pedagogical Application

Authors: Chia-Ling Hsieh, Kankanit Potikit

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Metadiscourse refers to linguistic resources employed by writers to organize text and interact with readers. While metadiscourse has received considerable attention within the field of discourse analysis, few studies have explored the use of metadiscourse in email, one of the most popular forms of computer-mediated communication. Furthermore, the diversity of cross-linguistic research required to uncover the influence of cultural factors on metadiscourse use is lacking. The present study compares metadiscourse markers employed in Chinese and Thai-language request emails with the purpose of discovering cross-cultural similarities and differences that are meaningful and applicable to foreign language teaching. The analysis is based on a corpus of 200 request emails: 100 composed in Chinese and 100 in Thai, with half of the emails from each language data set addressed to professors and the other half addressed to classmates. Adopting Hyland’s model as an analytical framework, two primary categories of metadiscourse are identified. Textual metadiscourse helps to create text coherence, while interpersonal metadiscourse functions to convey authorial stance. Results of the study make clear that both Chinese and Thai-language emails use significantly more interpersonal markers than textual markers, indicating that email, as a unique communicative medium, is characterized by high degrees of concision and interactivity. Users of both languages further deploy similar patterns in writing emails to recipients of different social statuses. Compared with emails addressed to classmates, emails addressed to professors are notably longer and include more transition and engagement markers. Nevertheless, cultural factors do play a role. Emails composed in Thai, for example, include more textual markers than those in Chinese, as Thai favors formal expressions and detailed explanations, while in contrast, emails composed in Chinese employ more interpersonal markers than those in Thai, since Chinese tends to emphasize recipient involvement and attitudinal warmth. These findings thereby demonstrate the combined effects of email as a communicative medium, social status, and cultural values on metadiscourse usage. The study concludes by applying these findings to pedagogical suggestions for teaching email writing to Chinese and Thai language learners based on similarities and differences in metadiscourse strategy between the two languages.

Keywords: discourse analysis, email, metadiscourse, writing instruction

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679 Allostatic Load as a Predictor of Adolescents’ Executive Function: A Longitudinal Network Analysis

Authors: Sipu Guo, Silin Huang

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Background: Most studies investigate the link between executive function and allostatic load (AL) among adults aged 18 years and older. Studies differed regarding the specific biological indicators studied and executive functions accounted for. Specific executive functions may be differentially related to allostatic load. We investigated the comorbidities of executive functions and allostatic load via network analysis. Methods: We included 603 adolescents (49.84% girls; Mean age = 12.38, SD age = 1.79) from junior high school in rural China. Eight biological markers at T1 and four executive function tasks at T2 were used to evaluate networks. Network analysis was used to determine the network structure, core symptoms, and bridge symptoms in the AL-executive function network among rural adolescents. Results: The executive functions were related to 6 AL biological markers, not to cortisol and epinephrine. The most influential symptoms were inhibition control, cognitive flexibility, processing speed, and systolic blood pressure (SBP). SBP, dehydroepiandrosterone, and processing speed were the bridges through which AL was related to executive functions. dehydroepiandrosterone strongly predicted processing speed. The SBP was the biggest influencer in the entire network. Conclusions: We found evidence for differential relations between markers and executive functions. SBP was a driver in the network; dehydroepiandrosterone showed strong relations with executive function.

Keywords: allostatic load, executive function, network analysis, rural adolescent

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678 Both Floristic Studies and Molecular Markers Are Necessary to Study of the Flora of a Region

Authors: Somayeh Akrami, Vali-Allah Mozaffarian, Habib Onsori

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The studied region in this research, watershed Kuhkamar river, is about 112.66 square kilometers, it is located between 45º 48' 9" to 45º 2' 20" N and 38º 34' 15" to 38º 40' 28" E. The gained results of the studies on flora combinations, proved 287 plant species in 190 genera and 51 families. Asteracea with 49 and Lamiaceae with 27 plant species are the major plant families. Among collected species one interesting plant was found and determined as a new record Anemone narcissiflora L. for flora of Iran. This plant is known as a complex species that shows intraspecific speciation and is classified into about 12 subspecies and 10 varieties in world. To identify the infraspecies taxons of this species, in addition to morphological characteristics, the use of appropriate molecular markers for the better isolation of the individuals were needed.

Keywords: Anemone narcissiflora, floristic Study, kuhkamar, molecular marker

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677 Impact of Nutritional Status on the Pubertal Transition in a Sample of Egyptian School Girls

Authors: Nayera E. Hassan, Salah Mostafa, Hamed Elkhayat, Kalled Hassan Sewidan, Sahar A. El-Masry, Manal Mouhamed Ali, Mones M. Abu Shady

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Pubertal growth is influenced by many factors including environmental and nutritional factors. Objective: To assess impact of nutritional status on pubertal staging, ovarian and uterine volumes among school girls. Method: Study was cross sectional and carried out on 1000 healthy school girls, aged 8-18 years selected randomly. They were categorized according to their ages into three groups: 8-12 years, 13-15 years and 16-18 years ±6 months, then according to their body mass index percentile to normal weight: (≥15-<85.), overweight (≥85-<95) and obese (≥95). All girls were subjected for physical, anthropometric (weight, height, body mass index), nutritional markers WAZ (weight/age Z score), HAZ (height/age Z score) and BMI-Z (body mass index Z score), pubertal assessment (Tanner stage) and pelvic transabdominal sonography (uterine and ovarian volumes). Results: Highly significant differences in ovarian and uterine volumes and nutritional markers (WAZ, HAZ and BMI-Z score) were detected among different grades of puberty in the two age groups (8-12 years, 13-15 years) coming in advance of obese girls (with increase of BMI); except HAZ in the second age group. Girls aged 16-18 years reached to final volume for the uterus and ovary with insignificant differences. Pubertal stage, ovarian and uterine sizes were highly significantly correlated with nutritional markers. Mean ages of onset: of puberty, menarche and complete puberty were, 11.65 + 1.84, 14.79 + 1.75 and 15.02 + 1.68 years respectively. Conclusion: Nutritional status has a crucial role in determining pubertal stage, ovarian and uterine volumes among Egyptian girls during the pubertal process.

Keywords: pubertal stage, nutritional markers, girls, ovarian and uterine volumes

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676 Identification and Validation of Co-Dominant Markers for Selection of the CO-4 Anthracnose Disease Resistance Gene in Common Bean Cultivar G2333

Authors: Annet Namusoke, Annet Namayanja, Peter Wasswa, Shakirah Nampijja

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Common bean cultivar G2333 which offers broad resistance for anthracnose has been widely used as a source of resistance in breeding for anthracnose resistance. The cultivar is pyramided with three genes namely CO-4, CO-5 and CO-7 and of these three genes, the CO-4 gene has been found to offer the broadest resistance. The main aim of this work was to identify and validate easily assayable PCR based co-dominant molecular markers for selection of the CO-4 gene in segregating populations derived from crosses of G2333 with RWR 1946 and RWR 2075, two commercial Andean cultivars highly susceptible to anthracnose. Marker sequences for the study were obtained by blasting the sequence of the COK-4 gene in the Phaseolus gene database. Primer sequence pairs that were not provided from the Phaseolus gene database were designed by the use of Primer3 software. PCR conditions were optimized and the PCR products were run on 6% HPAGE gel. Results of the polymorphism test indicated that out of 18 identified markers, only two markers namely BM588 and BM211 behaved co-dominantly. Phenotypic evaluation for reaction to anthracnose disease was done by inoculating 21days old seedlings of three parents, F1 and F2 populations with race 7 of Colletotrichum lindemuthianum in the humid chamber. DNA testing of the BM588 marker onto the F2 segregating population of the crosses RWR 1946 x G 2333 and RWR 2075 x G2333 further revealed that the marker BM588 co-segregated with disease resistance with co-dominance of two alleles of 200bp and 400bp, fitting the expected segregation ratio of 1:2:1. The BM588 marker was significantly associated with disease resistance and gave promising results for marker assisted selection of the CO-4 gene in the breeding lines. Activities to validate the BM211 marker are also underway.

Keywords: codominant, Colletotrichum lindemuthianum, MAS, Phaseolus vulgaris

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675 The Evaluation of Complete Blood Cell Count-Based Inflammatory Markers in Pediatric Obesity and Metabolic Syndrome

Authors: Mustafa M. Donma, Orkide Donma

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Obesity is defined as a severe chronic disease characterized by a low-grade inflammatory state. Therefore, inflammatory markers gained utmost importance during the evaluation of obesity and metabolic syndrome (MetS), a disease characterized by central obesity, elevated blood pressure, increased fasting blood glucose and elevated triglycerides or reduced high density lipoprotein cholesterol (HDL-C) values. Some inflammatory markers based upon complete blood cell count (CBC) are available. In this study, it was questioned which inflammatory marker was the best to evaluate the differences between various obesity groups. 514 pediatric individuals were recruited. 132 children with MetS, 155 morbid obese (MO), 90 obese (OB), 38 overweight (OW) and 99 children with normal BMI (N-BMI) were included into the scope of this study. Obesity groups were constituted using age- and sex-dependent body mass index (BMI) percentiles tabulated by World Health Organization. MetS components were determined to be able to specify children with MetS. CBC were determined using automated hematology analyzer. HDL-C analysis was performed. Using CBC parameters and HDL-C values, ratio markers of inflammation, which cover neutrophil-to-lymphocyte ratio (NLR), derived neutrophil-to-lymphocyte ratio (dNLR), platelet-to-lymphocyte ratio (PLR), lymphocyte-to-monocyte ratio (LMR), monocyte-to-HDL-C ratio (MHR) were calculated. Statistical analyses were performed. The statistical significance degree was considered as p < 0.05. There was no statistically significant difference among the groups in terms of platelet count, neutrophil count, lymphocyte count, monocyte count, and NLR. PLR differed significantly between OW and N-BMI as well as MetS. Monocyte-to HDL-C value exhibited statistical significance between MetS and N-BMI, OB, and MO groups. HDL-C value differed between MetS and N-BMI, OW, OB, MO groups. MHR was the ratio, which exhibits the best performance among the other CBC-based inflammatory markers. On the other hand, when MHR was compared to HDL-C only, it was suggested that HDL-C has given much more valuable information. Therefore, this parameter still keeps its value from the diagnostic point of view. Our results suggest that MHR can be an inflammatory marker during the evaluation of pediatric MetS, but the predictive value of this parameter was not superior to HDL-C during the evaluation of obesity.

Keywords: children, complete blood cell count, high density lipoprotein cholesterol, metabolic syndrome, obesity

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674 The Impacts of the Sit-Stand Workplace Intervention on Cardiometabolic Risk

Authors: Rebecca M. Dagger, Katy Hadgraft, Matthew Teggart, Peter Angell

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Background: There is a growing body of evidence that demonstrates the association between sedentary behaviour, cardiometabolic risk and all-cause mortality. Since full time working adults spend approximately 8 hours per day in the workplace, interventions to reduce sedentary behaviour at work may alleviate some of the negative health outcomes associated with sedentary behaviour. The aims of this pilot study were to assess the impacts of using a Sit-Stand workstation on markers of cardiometabolic health in a cohort of desk workers. Methods: Twenty eight participants were recruited and randomly assigned to a control (n=5 males, 9 females, mean age 37 years ± 9.4 years) or intervention group (n= 5 males, 9 females, mean age 42 years ± 12.7 years). All participants attended the labs on 2 occasion’s pre and post intervention, following baseline measurements the intervention participants had the Sit Stand Workstations (Ergotron, USA) installed for a 10 week intervention period. The Sit Stand workstations allow participants to stand or sit at their usual workstation and participants were encouraged to the use the desk in a standing position at regular intervals throughout the working day. Cardiometabolic risk markers assessed were body mass, body composition (using bio impedance analysis; Tanita, Tokyo), fasting blood Total Cholesterol (TC), lipid profiles (HDL-C, LDL-C, TC: HDL-C ratio), triglycerides and fasting glucose (Cholestech LDX), resting systolic and diastolic blood pressure and resting heart rate. ANCOVA controlling for baseline values was used to assess the group difference in changes in risk markers between pre and post intervention. Results: The 10 week intervention was associated with significant reductions in some cardiometabolic risk factors. There were significant group effects on change in body mass (F (1,25)=5.915, p<0.05), total body fat percentage (F(1,25)=12.615, p<0.01), total fat mass (F (1,25)=6.954, p<0.05), and systolic blood pressure (F (1,25)=5.012, p<0.05). There were no other significant group effects on changes in other cardiometabolic risk markers. Conclusion: This pilot study highlights the importance of reducing sedentary behaviour in the workplace for reduction in cardiometabolic risk markers. Further research is required to support these findings.

Keywords: sedentary behaviour, caridometabolic risk, evidence, risk makers

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673 Biopsy or Biomarkers: Which Is the Sample of Choice in Assessment of Liver Fibrosis?

Authors: S. H. Atef, N. H. Mahmoud, S. Abdrahman, A. Fattoh

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Background: The aim of the study is to assess the diagnostic value of fibrotest and hyaluronic acid in discriminate between insignificant and significant fibrosis. Also, to find out if these parameters could replace liver biopsy which is currently used for selection of chronic hepatitis C patients eligible for antiviral therapy. Study design: This study was conducted on 52 patients with HCV RNA detected by polymerase chain reaction (PCR) who had undergone liver biopsy and attending the internal medicine clinic at Ain Shams University Hospital. Liver fibrosis was evaluated according to the METAVIR scoring system on a scale of F0 to F4. Biochemical markers assessed were: alpha-2 macroglobulin (α2-MG), apolipoprotein A1 (Apo-A1), haptoglobin, gamma-glutamyl transferase (GGT), total bilirubin (TB) and hyaluronic acid (HA). The fibrotest score was computed after adjusting for age and gender. Predictive values and ROC curves were used to assess the accuracy of fibrotest and HA results. Results: For fibrotest, the observed area under curve for the discrimination between minimal or no fibrosis (F0-F1) and significant fibrosis (F2-F4) was 0.6736 for cutoff value 0.19 with sensitivity of 84.2% and specificity of 85.7%. For HA, the sensitivity was 89.5% and specificity was 85.7% and area under curve was 0.540 at the best cutoff value 71 mg/dL. Multi-use of both parameters, HA at 71 mg/dL with fibrotest score at 0.22 give a sensitivity 89.5%, specificity 100 and efficacy 92.3% (AUC 0.895). Conclusion: The use of both fibrotest score and HA could be as alternative to biopsy in most patients with chronic hepaitis C putting in consideration some limitations of the proposed markers in evaluating liver fibrosis.

Keywords: fibrotest, liver fibrosis, HCV RNA, biochemical markers

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672 RAPD Analysis of the Genetic Polymorphism in the Collection of Rye Cultivars

Authors: L. Petrovičová, Ž. Balážová, Z. Gálová, M. Wójcik-Jagła, M. Rapacz

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In the present study, RAPD-PCR was used to assess genetic diversity of the rye including landrances and new rye cultivars coming from Central Europe and the Union of Soviet Socialist Republics (SUN). Five arbitrary random primers were used to determine RAPD polymorphism in the set of 38 rye genotypes. These primers amplified altogether 43 different DNA fragments with an average number of 8.6 fragments per genotypes. The number of fragments ranged from 7 (RLZ 8, RLZ 9 and RLZ 10) to 12 (RLZ 6). DI and PIC values of all RAPD markers were higher than 0.8 that generally means high level of polymorphism detected between rye genotypes. The dendrogram based on hierarchical cluster analysis using UPGMA algorithm was prepared. The cultivars were grouped into two main clusters. In this experiment, RAPD proved to be a rapid, reliable and practicable method for revealing of polymorphism in the rye cultivars.

Keywords: genetic diversity, polymorphism, RAPD markers, Secale cereale L.

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671 From Genome to Field: Applying Genome Wide Association Study for Sustainable Ascochyta Blight Management in Faba Beans

Authors: Rabia Faridi, Rizwana Maqbool, Umara Sahar Rana, Zaheer Ahmad

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Climate change impacts agriculture, notably in Germany, where spring faba beans predominate. However, improved winter hardiness aligns with milder winters, enabling autumn-sown varieties. Genetic resistance to Ascochyta blight is vital for crop integration. Traditional breeding faces challenges due to complex inheritance. This study assessed 224 homozygous faba bean lines for Ascochyta resistance traits. To achieve h²>70%, 12 replicates were required (realized h²=87%). Genetic variation and strong trait correlations were observed. Five lines outperformed 29H, while three were highly susceptible. A genome-wide association study (GWAS) with 188 inbred lines and 2058 markers, including 17 guide SNP markers, identified 12 markers associated with resistance traits, potentially indicating new resistance genes. One guide marker (Vf-Mt1g014230-001) on chromosome III validated a known QTL. The guided marker approach complemented GWAS, facilitating marker-assisted selection for Ascochyta resistance. The Göttingen Winter Bean Population offers promise for resistance breeding.

Keywords: genome wide association studies, marker assisted breeding, faba bean, ascochyta blight

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670 Effect of Massage and Apium Graveolens Consumption on Liver Markers in Sedentary Women

Authors: M. Gholinjad Kordan, M. Ghasemi, S. Fazelifar

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Aim: liver plays a role in an astounding number of bodily functions: it helps with blood sugar regulation; metabolizes hormones; and traps and breaks down bacteria and toxins. Unfortunately, bodily demands can overwhelm your liver. Massage therapy can help by restoring healthy circulation and speeding the elimination of toxins. Apium graveolens has antioxidant property because of its flavonoid compounds. The purpose of this study was to determine Effect of massage and graveolens consumption on liver markers in women. Methodology: forty non-athlete male students were randomly divided into four groups: 1-control, 2-massage, 3-massage and graveolens and 4-graveolens. Amount of 5cc Blood sampling were obtained pre-test and post-test (after 12 weeks). Subjects received Russian massage for 8 weeks and 3 days a week for 20 minutes. Also, Complementary groups received the capsule supplement of graveolens after eating for twelve weeks and three times a day. Data was analyzed with One way ANOVA and tukey in p < 0/05. Results: Based on research findings, twelve weeks massage and graveolens consumption lead to a significant decrease in enzymes activity of ALT, AST and ALP in sedentary women. Conclusion: Results of This study shows that massage or graveolens consumption and especially combination of both can be used as desirable threpatic method in order to reduce or improve symptoms caused by excess weight, especially the liver damage.

Keywords: massage, graveolens, liver markers, sedentary women

Procedia PDF Downloads 268
669 Comparison Between Vegans and Omnivores on the Recovery of Delayed Onset Muscle Soreness in Young Females

Authors: Njeim Pressila, Hajj-Boutros Guy, Antony D. Karelis

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Background: Acute resistance exercise is associated with an elevated inflammation response, which could lead to delayed onset muscle soreness (DOMS). There is evidence that suggests that ingesting foods that have anti-inflammation properties may help reduce DOMS. A vegan diet has also been shown to be an anti-inflammatory diet which could, in turn, decrease DOMS. Objective and hypothesis: The purpose of the present study will be to compare markers of DOMS between vegans and omnivores after acute resistance exercise in young females. We hypothesize that vegans will have a better recovery of DOMS markers after a resistance exercise session compared to omnivores. Methods: Population: We will recruit30 vegans and 30 omnivores to participate in this study. Allvolunteers will follow either a vegan or an omnivore diet for at least 2 years. Anthropometric measurements, body composition, musclestrength (leg and chest press), markers of DOMS (swelling, pain, and stiffness), and dietary factors, as well as a wellness and anxiety questionnaire will be measured. All participants will also perform an acute resistance exercise session in order to induce DOMS. Pertinence: This project will give us a better understanding on the recovery process of vegans after a resistance training session and, as such, provide useful information to health professionals and athletes/coaches (kinesiologists and nutritionists)

Keywords: vgeans, omnivores, delayed onset muscle soreness, pain, stifness

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668 Molecular Analysis of Somaclonal Variation in Tissue Culture Derived Bananas Using MSAP and SSR Marker

Authors: Emma K. Sales, Nilda G. Butardo

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The project was undertaken to determine the effects of modified tissue culture protocols e.g. age of culture and hormone levels (2,4-D) in generating somaclonal variation. Moreover, the utility of molecular markers (SSR and MSAP) in sorting off types/somaclones were investigated. Results show that somaclonal variation is in effect due to prolonged subculture and high 2,4-D concentration. The resultant variation was observed to be due to high level of methylation events specifically cytosine methylation either at the internal or external cytosine and was identified by methylation sensitive amplification polymorphism (MSAP). Simple sequence repeats (SSR) on the other hand, was able to associate a marker to a trait of interest. These therefore, show that molecular markers can be an important tool in sorting out variation/mutants at an early stage.

Keywords: methylation, MSAP, somaclones, SSR, subculture, 2, 4-D

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667 Molecular Genetic Purity Test Using SSR Markers in Pigeon Pea

Authors: Rakesh C. Mathad, G. Y. Lokesh, Basavegowda

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In agriculture using quality seeds of improved varieties is very important to ensure higher productivity thereby food security and sustainability. To ensure good productivity, seeds should have characters as described by the breeder. To know whether the characters as described by the breeder are expressing in a variety such as genuineness or genetic purity, field grow out test (GOT) is done. In pigeon pea which is long durational crop, conducting a GOT may take very long time and expensive also. Since in pigeon pea flower character is a most distinguishing character from the contaminants, conducting a field grow out test require 120-130 days or till flower emergence, which may increase cost of storage and seed production. This will also delay the distribution of seed inventory to the pigeon pea growing areas. In this view during 2014-15 with financial support of Govt. of Karnataka, India, a project to develop a molecular genetic test for newly developed variety of pigeon pea cv.TS3R was commissioned at Seed Unit, UAS, Raichur. A molecular test was developed with the help SSR markers to identify pure variety from possible off types in newly released pigeon pea variety TS3R. In the investigation, 44 primer pairs were screened to identify the specific marker associated with this variety. Pigeon pea cv. TS3R could be clearly identified by using the primer CCM 293 based on the banding pattern resolved on gel electrophoresis and PCR reactions. However some of the markers like AHSSR 46, CCM 82 and CCM 57 can be used to test other popular varieties in the region like Asha, GRG-811 and Maruti respectively. Further to develop this in to a lab test, the seed sample size was standardized to 200 seeds and a grow out matrix was developed. This matrix was used to sample 12 days old leaves to extract DNA. The lab test results were validated with actual field GOT test results and found variations within the acceptable limit of 1%. This molecular method can now be employed to test the genetic purity in pigeon pea cv TS3R which reduces the time and can be a cheaper alternative method for field GOT.

Keywords: genuineness, grow-out matrix, molecular genetic purity, SSR markers

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666 Gold Nanoparticle Conjugated with Andrographolide Ameliorates Viper Venom-Induced Inflammatory Response and Organ Toxicity in Animal Model

Authors: Sourav Ghosh, Antony Gomes

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Since 1894 anti-snake venom serum (ASVS) is the only available treatment against snake envenomation, although there are many side effects and limitations. The need for a supportive treatment was felt for a long time to overcome the side effects and limitations of ASVS. Andrographolide conjugated with gold nanoparticle (A-GNP) has been found to antagonize viper venom-induced local damages. The present study was aimed to study the protective efficacy of A-GNP against Viper venom-induced inflammatory response and organ toxicity in animal model. Ethical clearance was obtained from animal experiments. Physico-chemical characterization of A-GNP was done by DLS (diameter and zeta potential), FE-SEM and XRD. Swiss albino male mice were divided into 4 groups: Gr.1-Sham control, Gr.2- Russell’s Viper venom (RVV) control, Gr.3- andrographolide treated and Gr.4- A-GNP treated. The 1/5th minimum lethal dose of RVV (500µg/kg, s.c.) was induced in animals of group 2, 3 & 4 animals, followed by treatment with andrographolide (100mg/kg, i.p.) and A-GNP (100mg/kg, i.v.) in group 3 & 4 animals, respectively. Blood was collected after 18 h, serum was prepared, and inflammatory markers (IL 1β, 6, 17a, 10, TNF α) and biochemical markers (AST, ACP, LDH, urea, creatinine) were assessed. Values were expressed as mean±SEM (n=4), one way ANOVA was done, P<0.05 was considered as statistically significant. DLS size showed the hydrodynamic diameter of A-GNP to be 230-260nm with polydispersity index of 0.103 and zeta potential was -18.32mV. XRD data confirmed the presence of crystalline gold in A-GNP, and FESEM indicated the presence of nearly spherical particle with size18-24nm.Treatment with A-GNP significantly decreased viper venom-induced proinflammatory markers (IL 1β, 6, 17, TNF α) increased anti-inflammatory markers (IL 10) and decreased organ toxicity markers (AST, ACP, LDH, urea, creatinine) in animal model. Venom neutralization efficacy of A-GNP was > andrographolide, which confirmed the increased efficacy of andrographolide after gold nanoparticle conjugation. Venom neutralization by A-GNP was due to anti-oxidant/anti-inflammatory activity of andrographolide, which showed increased efficacy after gold nanoparticle tagging. Thus, A-GNP may serve as a supportive therapy in snake-bite (against inflammatory response and organ toxicity) subject to further detail studies.

Keywords: andrographolide, gold nanoparticle, inflammatory response, organ toxicity, snake venom, snake venom neutralization, viper venom

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