Search results for: protein complementation assay

Authors: Anyaphat Srithanasuwan, Noppason Pangprasit, Montira Intanon, Phongsakorn Chuammitri, Witaya Suriyasathaporn, Ynte H. Schukken

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Streptococcus uberis is one of the most common mastitis-causing pathogens, with a wide range of intramammary infection (IMI) durations and pathogenicity. This study aimed to compare shared or unique virulence factor gene clusters distinguishing persistent and transient strains of S. uberis. A total of 139 S. uberis strains were isolated from three small-holder dairy herds with a high prevalence of S. uberis mastitis. The duration of IMI was used to categorize bacteria into two groups: transient and persistent strains with an IMI duration of less than 1 month and longer than 2 months, respectively. Six representative S. uberis strains, three from each group (transience and persistence) were selected for analysis. All transient strains exhibited multi-locus sequence types (MLST), indicating a highly diverse population of transient S. uberis. In contrast, MLST of persistent strains was available in an online database (pubMLST). Identification of virulence genes was performed using whole-genome sequencing (WGS) data. Differences in genomic size and number of virulent genes were found. For example, the BCA gene or alpha-c protein and the gene associated with capsule formation (hasAB), found in persistent strains, are important for attachment and invasion, as well as the evasion of the antimicrobial mechanisms and survival persistence, respectively. These findings suggest a genetic-level difference between the two strain types. Consequently, a comprehensive study of 139 S. uberis isolates will be conducted to perform an in-depth genetic assessment through WGS analysis on an Illumina platform.

Keywords: Streptococcus Uberis, mastitis, whole genome sequence, intramammary infection, persistent S. Uberis, transient s. Uberis

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Authors: Nail Nazarov, Alexandra Vyshtakalyuk, Vyacheslav Semenov, Irina Galyametdinova, Vladimir Zobov, Vladimir Reznik

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Among hepatic pyrimidine used as a means of stimulating protein synthesis and recovery of liver cells in her damaged toxic and infectious etiology. When an experimental toxic hepatitis hepatoprotective activity detected some pyrimidine derivatives. There are literature data on oksimetiluratcila hepatoprotective effect. For analogs of pyrimidine nucleobases - drugs Methyluracilum pentoxy and hepatoprotective effect of weakly expressed. According to the American scientists broad spectrum of biological activity, including hepatoprotective properties, have a 2,4-dioxo-5-arilidenimino uracils. Influenced Xymedon medicinal preparation (1- (beta-hydroxyethyl) -4,6-dimethyl-1,2-dihydro-2-oksopirimidin) developed as a means of stimulating the regeneration of tissue revealed increased activity of microsomal oxidases human liver. In studies on the model of toxic liver damage in rats have shown hepatoprotective effect xymedon and stimulating its impact on the recovery of the liver tissue. Hepatoprotective properties of the new compound in the series of pyrimidine derivatives L-ascorbate 1-(2-hydroxyethyl)-4,6-dimethyl-1,2-dihydropirimidine-2-one synthesized on the basis Xymedon preparation were firstly investigated on rats under the carbon tetrachloride action. It was shown the differences of biochemical parameters from the reference value and severity of structural-morphological liver violations decreased in comparison with control group under the influence of the compound injected before exposure carbon tetrachloride. Hepatoprotective properties of the investigated compound were more pronounced in comparison with Xymedon.

Keywords: hepatoprotectors, pyrimidine derivatives, toxic liver damage, xymedon

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Authors: Yohannes Kelifa, Gomathi Periasamy, Aman Karim

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Introduction: Diabetes mellitus has become a major public health and economical problem across the globe. Modern antidiabetic drugs have a number of limitations, and scientific investigation of traditional herbal remedies used for diabetes may provide novel leads for the development of new antidiabetic drugs that can be used as alternative or complementary to available antidiabetic allopathic medications. Though Myrica salicifolia Hochst. ex A. Rich. is used for the management of diabetes in Ethiopian traditional medicine, there was no previous scientific evidence about its antidiabetic effect to the authors’ knowledge. This study was undertaken to evaluate the antidiabetic activity the root extracts of Myrica salicifolia in streptozotocin (STZ)-induced diabetic mice. Methods: Experimental diabetes was induced by intraperitoneal administration of STZ (150 mg/kg) in male mice. Diabetic mice were treated with oral doses of M. salicifolia root extracts at 200, 400 and 600 mg/kg, and its fractions (chloroform, ethyl acetate, n-butanol and aqueous) at a dose of 400 mg/kg daily for 15 days. Fasting blood glucose level (BGL) was measured at 0, 5th,10th, and 15th day. The free radical scavenging activity of the crude extract was determined using in vitro by DPPH assay. The statistical significance was assessed by one-way ANOVA, followed by Tukey’s multiple comparison tests. Results were considered significant when p < 0.05. Results: Daily administration of the M. salicifolia 80% methanol root extracts (at three different doses (200, 400 and 600 mg/kg) significantly (p < 0.05, p < 0.01 and p < 0.001) reduced fasting BGL compared with diabetic control. The aqueous and butanol fractions at a dose of 400 mg/kg resulted in maximum reduction of fasting BGL by 42.39%, and 52.13%, respectively at the 15th day in STZ-induced diabetic mice. Free radical scavenging activity of the 80% methanol extract of M. salicifolia was comparable to ascorbic acid. The IC50 values of the crude extract and ascorbic acid (a reference compound) were found to be 4.54 μg/ml and 4.39 μg/ml, respectively. Conclusion: These findings demonstrated that the methanolic extracts of M. salicifolia root and its fractions (n-butanol and aqueous) exhibit a significant antihyperglycemic activity in STZ-induced diabetic mice. Furthermore, the result of the present study indicates that M. salicifolia root extract is a potential source of natural antioxidants.

Keywords: antidiabetic, diabetes mellitus, DPPH, mice, Myrica salicifolia, streptozotocin

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Authors: Larisa Gheber

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Mitosis is an essential process by which duplicated genetic information is transmitted from mother to daughter cells. Incorrect chromosome segregation during mitosis can lead to genetic diseases, chromosome instability and cancer. This process is mediated by a dynamic microtubule-based intracellular structure, the mitotic spindle. One of the major factors that govern the mitotic spindle dynamics are the kinesin-5 biological nano motors that were believed to move unidirectionally on the microtubule filaments, using ATP hydrolysis, thus performing essential functions in mitotic spindle dynamics. Surprisingly, several reports from our and other laboratories have demonstrated that some kinesin-5 motors are bi-directional: they move in minus-end direction on the microtubules as single-molecules and can switch directionality under a number of conditions. These findings broke a twenty-five-years old dogma regarding kinesin directionality (1, 2). The mechanism of this bi-directional motility and its physiological significance remain unclear. To address this unresolved problem, we apply an interdisciplinary approach combining live cell imaging, biophysical single molecule, and structural experiments to examine the activity of these motors and their mutated variants in vivo and in vitro. Our data shows that factors such as protein phosphorylation (3, 4), motor clustering on the microtubules (5, 6) and structural elements (7, 8) regulate the bi-directional motility of kinesin motors. We also show, using Cryo-EM, that bi-directional kinesin motors obtain non-canonical microtubule binding, which is essential to their special motile properties and intracellular functions. We will discuss the implication of these findings to mechanism bi-directional motility and physiological roles in mitosis.

Keywords: mitosis, cancer, kinesin, microtubules, biochemistry, biophysics

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Authors: S. Noreen, S. Ahmad

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Salinity is one of the most important environmental factors that limit the production of crop plants to the greatest proportion than any other ones. Salt-induced changes in growth, pigment concentration, water status, malondialdehydes (MDA) and H₂O₂ content, enzymatic and non-enzymatic antioxidants, Na⁺, K⁺ content and yield attributes were examined in the glasshouse on ten pea (Pisum Sativum L.) accessions, namely ‘13240’, ‘18302’, ‘19666’, ‘19700’, ‘19776’, ‘19785’, ‘19788’, ‘20153’, ‘20155’, ‘26719’ were subjected to non-stress (0 mM NaCl) and salt stress (100 mM and150 mM NaCl) in pots containing sand medium. The results showed that salt stress at level150 mM substantially reduced biomass production, leaf water status, pigment concentration (chlorophyll ‘a’, ‘b’, ‘carotenoid content’ total chlorophyll), K⁺ content, quantum yield and yield attributes as compared to plants treated with 100 mM NaCl. Antioxidant enzymes, Catalase (CAT), Peroxidase (POD), Superoxide dismutase (SOD) and Ascorbate peroxidase (APX), proline content, total soluble protein, total amino acids, Malondialdehyde content (MDA), Hydrogen peroxide (H₂O₂) content and Na⁺ uptake markedly enhanced due to the influence of salt stress. On the basis of analyses (expressed as percent of control), of 10 accessions of pea plant, two were ranked as salt tolerant namely (‘19666’, ‘20153’), four were moderately tolerant namely (‘19700’, ‘19776’, ‘19785’, ‘20155’), and three were salt sensitive namely (‘13240’, ‘18302’, ‘26719’) at 150 mM NaCl level.

Keywords: antioxidant enzymes, ion uptake, pigment concentration, salt stress, yield attributes

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Authors: Uraib Sharaha, Ahmad Salman, Eladio Rodriguez-Diaz, Elad Shufan, Klaris Riesenberg, Irving J. Bigio, Mahmoud Huleihel

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Antimicrobial drugs have an important role in controlling illness associated with infectious diseases in animals and humans. However, the increasing resistance of bacteria to a broad spectrum of commonly used antibiotics has become a global health-care problem. Rapid determination of antimicrobial susceptibility of a clinical isolate is often crucial for the optimal antimicrobial therapy of infected patients and in many cases can save lives. The conventional methods for susceptibility testing like disk diffusion are time-consuming and other method including E-test, genotyping are relatively expensive. Fourier transform infrared (FTIR) microscopy is rapid, safe, and low cost method that was widely and successfully used in different studies for the identification of various biological samples including bacteria. The new modern infrared (IR) spectrometers with high spectral resolution enable measuring unprecedented biochemical information from cells at the molecular level. Moreover, the development of new bioinformatics analyses combined with IR spectroscopy becomes a powerful technique, which enables the detection of structural changes associated with resistivity. The main goal of this study is to evaluate the potential of the FTIR microscopy in tandem with machine learning algorithms for rapid and reliable identification of bacterial susceptibility to antibiotics in time span of few minutes. The bacterial samples, which were identified at the species level by MALDI-TOF and examined for their susceptibility by the routine assay (micro-diffusion discs), are obtained from the bacteriology laboratories in Soroka University Medical Center (SUMC). These samples were examined by FTIR microscopy and analyzed by advanced statistical methods. Our results, based on 550 E.coli samples, were promising and showed that by using infrared spectroscopic technique together with multivariate analysis, it is possible to classify the tested bacteria into sensitive and resistant with success rate higher than 85% for eight different antibiotics. Based on these preliminary results, it is worthwhile to continue developing the FTIR microscopy technique as a rapid and reliable method for identification antibiotic susceptibility.

Keywords: antibiotics, E. coli, FTIR, multivariate analysis, susceptibility

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Authors: Aurea C. L. Lacerda, Paulo R. H. Moreno, Bruna M. P. Vianna, Cristina H. R. Serra, Airton Martin, André R. Baby, Vladi O. Consiglieri, Telma M. Kaneko

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The modified Franz cell is the most widely used model for in vitro permeation studies, however it still presents some disadvantages. Thus, some alternative methods have been developed such as Skin PAMPA, which is a bio- artificial membrane that has been applied for skin penetration estimation of xenobiotics based on HT permeability model consisting. Skin PAMPA greatest advantage is to carry out more tests, in a fast and inexpensive way. The membrane system mimics the stratum corneum characteristics, which is the primary skin barrier. The barrier properties are given by corneocytes embedded in a multilamellar lipid matrix. This layer is the main penetration route through the paracellular permeation pathway and it consists of a mixture of cholesterol, ceramides, and fatty acids as the dominant components. However, there is no consensus on the membrane composition. The objective of this work was to compare the performance among different bio-artificial membranes for studying the permeation in skin PAMPA system. Material and methods: In order to mimetize the lipid composition`s present in the human stratum corneum six membranes were developed. The membrane composition was equimolar mixture of cholesterol, ceramides 1-O-C18:1, C22, and C20, plus fatty acids C20 and C24. The membrane integrity assay was based on the transport of Brilliant Cresyl Blue, which has a low permeability; and Lucifer Yellow with very poor permeability and should effectively be completely rejected. The membrane characterization was performed using Confocal Laser Raman Spectroscopy, using stabilized laser at 785 nm with 10 second integration time and 2 accumulations. The membrane behaviour results on the PAMPA system were statistically evaluated and all of the compositions have shown integrity and permeability. The confocal Raman spectra were obtained in the region of 800-1200 cm-1 that is associated with the C-C stretches of the carbon scaffold from the stratum corneum lipids showed similar pattern for all the membranes. The ceramides, long chain fatty acids and cholesterol in equimolar ratio permitted to obtain lipid mixtures with self-organization capability, similar to that occurring into the stratum corneum. Conclusion: The artificial biological membranes studied for Skin PAMPA showed to be similar and with comparable properties to the stratum corneum.

Keywords: bio-artificial membranes, comparison, confocal Raman, skin PAMPA

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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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Authors: Jillian M. Hagel, Joseph E. Tucker, Peter J. Facchini

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With the aim of establishing a holistic approach for the treatment of central nervous system (CNS) disorders, we are pursuing a drug development program rapidly progressing through discovery and characterization phases. The drug candidates identified in this program are referred to as neuroplastogens owing to their ability to mediate neuroplasticity, which can be beneficial to patients suffering from anxiety, depression, or posttraumatic stress disorder. These and other related neuropsychiatric conditions are associated with the onset of neuronal atrophy, which is defined as a reduction in the number and/or productivity of neurons. The stimulation of neuroplasticity results in an increase in the connectivity between neurons and promotes the restoration of healthy brain function. We have synthesized a substantial catalogue of proprietary indolethylamine derivatives based on the general structures of serotonin (5-hydroxytryptamine) and psychedelic molecules such as N,N-dimethyltryptamine (DMT) and psilocin (4-hydroxy-DMT) that function as neuroplastogens. A primary objective in our screening protocol is the identification of derivatives associated with a significant reduction in hallucination, which will allow administration of the drug at a dose that induces neuroplasticity and triggers other efficacious outcomes in the treatment of targeted CNS disorders but which does not cause a psychedelic response in the patient. Both neuroplasticity and hallucination are associated with engagement of the 5HT2A receptor, requiring drug candidates differentially coupled to these two outcomes at a molecular level. We use novel and proprietary artificial intelligence algorithms to predict the mode of binding to the 5HT2A receptor, which has been shown to correlate with the hallucinogenic response. Hallucination is tested using the mouse head-twitch response model, whereas mouse marble-burying and sucrose preference assays are used to evaluate anxiolytic and anti-depressive potential. Neuroplasticity is assays using dendritic outgrowth assays and cell-based ELISA analysis. Pharmacokinetics and additional receptor-binding analyses also contribute the selection of lead candidates. A summary of the program is presented.

Keywords: neuroplastogen, non-hallucinogenic, drug development, anxiety, depression, PTSD, indolethylamine derivatives, psychedelic-inspired, 5-HT2A receptor, computational chemistry, head-twitch response behavioural model, neurite outgrowth assay

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Authors: Hussein M. Elmehdi

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In this paper, we present the results of our recent experiments done to examine the effect of air bubbles, which were introduced to bio-samples during preparation, on the rheological properties of soft biological materials. To effectively achieve this, we three samples each prepared with differently. Our soft biological systems comprised of three types of flour dough systems made from different flour varieties with variable protein concentrations. The samples were investigated using ultrasonic waves operated at low frequency in transmission mode. The sample investigated included dough made from bread flour, wheat flour and all-purpose flour. During mixing, the main ingredient of the samples (the flour) was transformed into cohesive dough comprised of the continuous dough matrix and air pebbles. The rheological properties of such materials determine the quality of the end cereal product. Two ultrasonic parameters, the longitudinal velocity and attenuation coefficient were found to be very sensitive to properties such as the size of the occluded bubbles, and hence have great potential of providing quantitative evaluation of the properties of such materials. The results showed that the magnitudes of the ultrasonic velocity and attenuation coefficient peaked at optimum mixing times; the latter of which is taken as an indication of the end of the mixing process. There was an agreement between the results obtained by conventional rheology and ultrasound measurements, thus showing the potential of the use of ultrasound as an on-line quality control technique for dough-based products. The results of this work are explained with respect to the molecular changes occurring in the dough system as the mixing process proceeds; particular emphasis is placed on the presence of free water and bound water.

Keywords: ultrasound, soft biological materials, velocity, attenuation

Procedia PDF Downloads 244

Authors: E. Riani, T. T. Irawadi, S. Nurjanah, K. Syamsu, E. G. Said, Suprihatin, M. R. Cordova

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Especially coastal, Indonesian and Chinese communities have utilized sandfish to improve reproduction quality of men. This study aimed to examine the nutrition on sandfish flesh that has the potency to improve reproduction quality of men. The materials used were sandfish with weight of 200-500 g, and then analysis of proximate, analysis of amino acid, analysis of fatty acid and analysis of mineral contained in the sandfish were performed. The results showed that protein content (39.96%) was the main component of the flesh; the carbohydrate and fat were 25.43% and 4.18%, respectively. Sandfish powder contains several essential amino acids and nonessential amino acids. Nine of ten amino acids needed by human body are contained in sandfish powder, i.e. arginine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, tryptophan, threonine and valine; only tryptophan that are not contained in sandfish powder. Sandfish powder contains saturated fatty acid kaproat, kaprilat, kaprat, laurat, miristat, stearat, arakhidat and behenat; monosaturated fatty acids (MUFA) and polyunsaturated fatty acids (PUFA). MUFA is composed of fatty acid oleat, while PUFA is composed fatty acid omega 3 (linonenat, eicosapentaenoic acid and docosahexaenoic acid) and omega 6 (linoleat and arakhidonat). The minerals contained in sandfish powder are macrominerals and microminerals. Based on the findings, the nutrition in sandfish powder has a good potency to improve reproduction of men, especially PUFA for the maturation of spermatozoa, zinc for production function and spermatogenesis, motility of spermatozoa, acromoson reaction; Mg for transformation of genetic information and motility of spermatozoa; calcium for spermatogenesis, capacity and fertilization of spermatozoa. Thus, sandfish flesh powder has the potency to improve reproduction quality of men.

Keywords: sandfish flesh powder, nutrition, reproduction quality, men

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Authors: Paria Baharvand, Mohammad Reza Abbasi, Soha Namazi

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Introduction: Uremic pruritus (UP) is one of the major complaints in hemodialysis patients. The aim of this randomized clinical trial study was the evaluation of the anti-pruritus effect of Melatonin on hemodialysis patients with UP. Methods: This multi-centered double, blinded randomized clinical trial study was conducted on hemodialysis patients with UP that referred to the dialysis department of Imam Khomeini Hospital, Labbafinezhad Hospital and Ghiasi Hospital in Tehran, Iran, in 2018. Forty participants were enrolled and block randomization was performed by using a randomized list and divided into 2 groups of twenty patients. Evaluation of the 12-Item Pruritus Severity Scale (12-PSS), Visual analog scale (VAS), and Calculation of the affected body surface area for pruritus extent (BSA %) was performed every week. The Melatonin group received a 10 mg soft gel capsule, and the placebo group received a soft gel capsule placebo after dinner. For statistical analysis used by SPSS version 21(IRCT20180714040462N1). Results: 38 hemodialysis patients enrolled in this study. The mean age in the placebo group and in the Melatonin group was 55.88 ± 11.70 and 54.52 ± 13.00 years (p> 0.05). Also, 46% of all participants were female. Aspartate transaminase, alanine transaminase and C-reactive protein have significant differences between the two groups (p< 0.05). VAS score reduction in the two groups had a statistically significant (CI 95% = -1.71, P= 0.015(.The results of BSA% showed Melatonin had a significant effect on the decrease of the pruritus extent compared to placebo (CI 95% = -22.82, P= 0.001(. Conclusion: According to the results of this study and the preliminary effects of Melatonin on uremic pruritus, we suggest performing advanced clinical trials studies.

Keywords: Melatonin, Uremic Pruritus, hemodialysis, anti-pruritus

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Authors: Aydan Atalar, Nurcan Cetinkaya

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Corn stover is leftover of the leaves, stalk, husks and tassels in the field after harvesting the grain combined. Corn stover is a low-quality roughage but has mostly been used as roughage source for feeding ruminant animals in developing countries including Turkey; however, it can also be used to make biofuels as in developed countries. The objectives of the present study were to improve the digestibility of corn stover by the treatment of white rod fungus mainly Pleurotus osteritus (PO), Pleurotus eryingii (PE) and Lantinula edudes (LE) at different incubation times and also to determine the most effective fungus and incubation time to prepare fermeted corn stover for ruminant nutrition. The choped corn stover was treated with PO, PE and LE and incubated for 10, 20, 30 and 40 days in incubator at 26 0C. After each incubation time dry matter(DM), organic matter(OM), crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF), neutral detergent lignin (ADL), in-vitro true dry matter digestibility (IVTDMD) and organic matter digestibility (IVTOMD) were determined. The mean IVTDMD and IVTOMD levels were increased by PO, PE and LE treatments in increasing order of incubation times. The obtained IVTDM values were 59.45, 60.51, 60.82 and 60.18 %; 59.45, 70.55, 67.18 and 66.96 %; 59.45, 70.55, 67.18 and 66,96 %; 59.45, 74.90, 69.18 % ; 59.45, 76.50, 71.24 and 73.04 for control, PO, PE and LE treatments at 0, 10, 20, 30 and 40 days incubation times respectively. The obtained IVTOMD values were 56.45,60.26,60.82and 60.18 %; 56.45, 68.70, 67.18 and 66.96 %; 56.45, 71.26, 69.18 and 69.28 %; 56.45, 73.23, 71.24 and 73.04 % for control, PO, PE and LE treatments at 0, 10, 20, 30 and 40 days incubation times respectively. The most effective fungus was PO and the incubation time was 30 days. In conclusion, PO treatment of corn stover with 30 days incubation may be used to prepare fermented corn stover for ruminant nutrition.

Keywords: biological treatment, corn stover, digestibility, Lantinula edudes, Pleurotus eryingii, Pleurotus osteritus

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Authors: Damaris Estrella Castillo, Zacil ha Vilchis Zapata, Leydi Peraza Gómez

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Hearing loss is an etiologically heterogeneous condition, where almost 60% is genetic in origin, 20% is due to environmental factors, and 20% have unknown causes. However, it is now known that the gene, GJB2, which encodes the connexin 26 protein, accounts for a large percentage of non-syndromic genetic hearing loss, and variants in this gene have been identified to be a common cause of hereditary hearing loss in many populations. The literature reports that the etiology in deafness helps improve family functioning but low-income countries this is difficult. Therefore, it is difficult to contribute the right of families to know about the genetic risk in future pregnancies as well as determining the certainty of being a carrier or affected. In order to assess the impact of genetic counseling and the functionality, 100 families with at least one child with profound hearing loss, were evaluated by specialists in audiology, clinical genetics and psychology. Targeted mutation analysis for one of the two known large deletions of upstream of GJB2/GJB6 gene (35delG; and including GJB2 regulatory sequences and GJB6) were performed in patients with diagnosis of non-syndromic hearing loss. Genetic counseling was given to all parents and primary caregivers, and APGAR family test was applied before and after the counseling. We analyzed a total of 300 members (children, parents) to determine the presence of the GJB2 gene mutation. Twelve patients (carriers and affected) were positive for the mutation, from 5 different families. The subsequent family APGAR testing and genetic counseling, showed that 14% perceived their families as functional, 62 % and 24 % moderately functional dysfunctional. This shows the importance of genetic counseling in the perception of family function that can directly impact the quality of life of these families.

Keywords: family dynamics, deafness, APGAR, counseling

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Authors: Amanat Ali, Muhammad S. Al-Siyabi, Mostafa I. Waly, Hashem Al-Kilani

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In a cross-sectional research design, we assessed the nutrient intake, nutritional status, nutritional knowledge and dietary habits of Sultan Qaboos University (SQU) student athletes. A total of 71 (49 male and 22 female) student athletes with a mean age of 21.0 ± 1.81 and 19.32 ± 0.72 years and body mass index (BMI) of 22.51 ± 1.98 and 20.34 ± 2.97 kg/m2 for male and female respectively, participated in this study. A study questionnaire consisting of 2 sections was distributed to the participants. Section I included 18 questions regarding the demographic information, whereas the Section II consisted of 20 questions regarding the nutrition knowledge. The dietary intake of participants was collected by using a 7-days food diary identifying the frequency as well as the variety of food consumption. Significant differences (P < 0.05) were observed in the main sources of nutrition information used by the male and female athletes. Male athletes mainly had most of the nutrition information from friends (17%) whereas female athletes relied mainly on the family (20%). More female athletes (20%) were using TV as a source of nutrition information as compared to male athletes (15%). Both male and female athletes had the minimum nutrition information from dietitians and physicians. Significant (P < 0.05) differences were also observed in the nutritional knowledge and dietary habits scores of male and female athletes, which were 57 % and 49 %, respectively. Male athletes were classified to have fair nutritional knowledge and dietary habits, whereas the female athletes had poor nutritional knowledge and dietary habits. The average daily energy intake of male athletes was 2595 ± 358 kcal/day. Carbohydrate, fat, and protein contributed 64%, 22%, and 14%, of the total energy intake for the male athletes, respectively. The energy and macronutrients intake of male athletes was within the recommended dietary intake. The results indicated some gaps in the nutritional knowledge of SQU student athletes and suggest that there is a need for developing strategies in counseling and teaching the athletes to improve their nutritional knowledge and dietary habits.

Keywords: nutrient assessment, nutritional knowledge, dietary habits, Omani University athletes

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Authors: Muhammad Adeel Arshad, Faiz-Ul Hassan, Yanfen Cheng

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According to FAO, enteric methane (CH4) production is about 44% of all greenhouse gas emissions from the livestock sector. Ruminants produce CH4 as a result of fermentation of feed in the rumen especially from roughages which yield more CH4 per unit of biomass ingested as compared to concentrates. Efficient ruminal fermentation is not possible without abating CO2 and CH4. Methane abatement strategies are required to curb the predicted rise in emissions associated with greater ruminant production in future to meet ever increasing animal protein requirements. Ecology of ruminal methanogenesis and avenues for its mitigation can be identified through various genomic and transcriptomic techniques. Programs such as Hungate1000 and the Global Rumen Census have been launched to enhance our understanding about global ruminal microbial communities. Through Hungate1000 project, a comprehensive reference set of rumen microbial genome sequences has been developed from cultivated rumen bacteria and methanogenic archaea along with representative rumen anaerobic fungi and ciliate protozoa cultures. But still many species of rumen microbes are underrepresented especially uncultivable microbes. Lack of sequence information specific to the rumen's microbial community has inhibited efforts to use genomic data to identify specific set of species and their target genes involved in methanogenesis. Metagenomic and metatranscriptomic study of entire microbial rumen populations offer new perspectives to understand interaction of methanogens with other rumen microbes and their potential association with total gas and methane production. Deep understanding of methanogenic pathway will help to devise potentially effective strategies to abate methane production while increasing feed efficiency in ruminants.

Keywords: Genome sequences, Hungate1000, methanogens, ruminal fermentation

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Authors: Tanveer Hussain, Misbah Hussain, Masroor Ellahi Babar, Muhammad Traiq Pervez, Fiaz Hussain, Sana Zahoor, Rashid Saif

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Interleukin 2 (IL-2) is a cytokine which is produced by activated T cells, play important role in immune response against antigen. It act in both autocrine and paracrine manner. It can stimulate B cells and various other phagocytic cells like monocytes, lymphokine-activated killer cells and natural killer cells. Acting in autocrine fashion, IL-2 protein plays a crucial role in proliferation of T cells. IL-2 triggers the release of pro and anti- inflammatory cytokines by activating several pathways. In present study, exon 1 of IL-2 gene of four local Pakistani breeds (Dera Din Panah, Beetal, Nachi and Kamori) from two provinces was amplified by using reported Ovine IL-2 primers, yielding PCR product of 501 bp. The sequencing of all samples was done to identify the polymorphisms in amplified region of IL-2 gene. Analysis of sequencing data resulted in identification of one novel nucleotide substitution (T→A) in amplified non-coding region of IL-2 gene. Comparison of IL-2 gene sequence of all four breeds with other goat breeds showed high similarity in sequence. While phylogenetic analysis of our local breeds with other mammals showed that IL-2 is a variable gene which has undergone many substitutions. This high substitution rate can be due to the decreased or increased changed selective pressure. These rapid changes can also lead to the change in function of immune system. This pioneering study of Pakistani goat breeds urge for further studies on immune system of each targeted breed for fully understanding the functional role of IL-2 in goat immunity.

Keywords: interleukin 2, mutational analysis, phylogeny, goat breeds, Pakistan

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Authors: Divyansh Goel, Varun Jain

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Classical economics works on the principle that people are rational and analytical in their decision making and their choices fall in line with the most suitable option according to the dominant strategy in a standard game theory model. This model has failed at many occasions in estimating the behavior and dealings of rational people, giving proof of some other underlying heuristics and cognitive biases at work. This paper probes into the study of these factors, which fall under the umbrella of behavioral economics and through their medium explore the solution to a problem which a lot of nations presently face. There has long been a wide disparity in the number of people holding favorable views on organ donation and the actual number of people signing up for the same. This paper, in its entirety, is an attempt to shape the public policy which leads to an increase the number of organ donations that take place and close the gap in the statistics of the people who believe in signing up for organ donation and the ones who actually do. The key assumption here is that in cases of cognitive dissonance, where people have an inconsistency due to conflicting views, people have a tendency to go with the default choice. This tendency is a well-documented cognitive bias known as the status quo bias. The research in this project involves an assay of mandated choice models of organ donation with two case studies. The first of an opt-in system of Germany (where people have to explicitly sign up for organ donation) and the second of an opt-out system of Austria (every citizen at the time of their birth is an organ donor and has to explicitly sign up for refusal). Additionally, there has also been presented a detailed analysis of the experiment performed by Eric J. Johnson and Daniel G. Goldstein. Their research as well as many other independent experiments such as that by Tsvetelina Yordanova of the University of Sofia, both of which yield similar results. The conclusion being that the general population has by and large no rigid stand on organ donation and are gullible to status quo bias, which in turn can determine whether a large majority of people will consent to organ donation or not. Thus, in our paper, we throw light on how governments can use status quo bias to drive positive social change by making policies in which everyone by default is marked an organ donor, which will, in turn, save the lives of people who succumb on organ transplantation waitlists and save the economy countless hours of economic productivity.

Keywords: behavioral economics, game theory, organ donation, status quo bias

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Authors: Aldi Nel, Cliff L. W. Jones, Justin O. G. Kemp, Peter J. Britz

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Kelp Ecklonia maxima is included in formulated abalone feeds in South Africa, but its effect on abalone growth, feed utilisation efficiency and gut-bacterial communities has not previously been investigated. An eight-month on-farm growth trial with sub-adult Haliotis midae (~43 mm shell length) fed graded levels of kelp in formulated feeds was conducted. Kelp inclusion (0.44–3.54 % of pellet dry mass) promoted faster growth (65.7 – 74.5 % total mass gain), with better feed and protein conversions (FCR: 1.4 – 1.8; PER 2.3 – 2.7), compared to abalone fed the non-supplemented feed (52.3% total mass gain; FCR: 2.1; PER 1.9; p < 0.001). The gut-bacterial communities of abalone fed kelp-supplemented feed (0.88 % of pellet dry mass) were subsequently compared with that of abalone fed a non-supplemented control diet. Abalone gut-bacterial DNA was sequenced using 16S rRNA pyrosequencing and sequences were clustered into operational taxonomic units (OTUs) at a 97 % similarity level. A supplementary 16S rRNA denaturing gradient gel electrophoresis (DGGE) analysis was conducted. The dominant OTUs differed in terms of their relative abundances, with that of an autochthonous Mollicutes strain being significantly higher (p = 0.03) in the guts of abalone fed kelp-supplemented feed. The DGGE band patterns displayed a higher within-group variability of dominant bacterial strains for abalone fed the control diet, suggesting that dietary inclusion of kelp, which is rich in fermentable polysaccharides, promotes a balanced gut-bacterial community. This may contribute to the better feed utilisation and growth in abalone fed kelp-supplemented feeds.

Keywords: abfeed, digestion, macroalgae, mariculture

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Authors: Naseem Fatima, A. N. Srivastava, Tasleem Raza, Vijay Kumar

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Introduction: Carcinoma gallbladder is third most common gastrointestinal lethal disease with the highest incidence and mortality rate among women in Northern India. Scientists have found several risk factors that make a person more likely to develop gallbladder cancer; among these risk factors, deregulation of miRNAs has been demonstrated to be one of the most crucial factors. The changes in the expression of specific miRNA genes result in the control of inflammation, cell cycle regulation, stress response, proliferation, differentiation, apoptosis and invasion thus mediate the process in tumorgenesis. The aim of this study was to investigate the role of MiRNA-335 and may as a molecular marker in early detection of gallbladder cancer in suspected cases. Material and Methods: A total of 20 consecutive patients with gallbladder cancer aged between 30-75 years were registered for the study. Total RNA was extracted from tissue by using the mirVANA MiRNA isolation Kit according to the manufacturer’s protocol. The MiRNA- 335 and U6 snRNA-specific cDNA were reverse-transcribed from total RNA using Taqman microRNA reverse-transcription kit according to the manufacturer’s protocol. TaqMan MiRNA probes hsa-miR-335 and Taqman Master Mix without AmpEase UNG, Individual real-time PCR assays were performed in a 20 μL reaction volume on a Real-Time PCR system (Applied Biosystems StepOnePlus™) to detect MiRNA-335 expression in tissue. Relative quantification of target MiRNA expression was evaluated using the comparative cycle threshold (CT) method. The correlation was done in between cycle threshold (CT Value) of target MiRNA in gallbladder cancer with respect to non-cancerous Cholelithiasis gallbladder. Each sample was examined in triplicate. The Newman-Keuls Multiple Comparison Test was used to determine the expression of miR-335. Results: MiRNA335 was found to be significantly downregulated in the gallbladder cancer tissue (P<0.001), when compared with non-cancerous Cholelithiasis gallbladder cases. Out of 20 cases, 75% showed reduced expression of MiRNA335, were at last stage of disease with low overall survival rate and remaining 25% were showed up-regulated expression of MiRNA335 with high survival rate. Conclusion: The present study showed that reduced expression of MiRNA335 is associated with the advancement of the disease, and its deregulation may provide important clues to understanding it as a prognostic marker and opportunities for future research.

Keywords: carcinoma gallbladder, downregulation, MiRNA-335, RT-PCR assay

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Authors: Leila M. Carvalho, Maria Erica S. Oliveira, Arnoud C. Neto, Elza I. Ida, Massami Shimokomaki, Marta S. Madruga

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The quality of broiler breast meat is changing as a result of the continuing emphasis on genetic selection for a more efficient meat production. Breast meat has been classified as PSE (pale, soft, exudative), DFD (dark, firm, dry) and normal color meat, and recently a third group has emerged: the so-called PFN (pale, firm, non-exudative) meat. This classification was based on pH, color and functional properties. The aim of this work was to confirm the existence of PFN and PSE meat by biochemical characterization and functional properties. Twenty four hours of refrigerated fillet, Pectoralis major, m. samples (n= 838) were taken from Cobb flocks 42-48 days old, obtained in Northeastern Brazil tropical region, the Northeastern, considered to have only dry and wet seasons. Color (L*), pH, water holding capacity (WHC), values were evaluated and compared with PSE group samples. These samples were classified as Normal (465.8), PSE meat (L*≥53; pH<5.8) and PFN (L*≥53; pH>5.8). The occurrence of control meat, PSE and PFN was 69.09%, 11.10% and 19.81%, respectively. Samples from PFN presented 4.0-5.0% higher WHC in relation to PSE meat and similar to control group. These results are explained by the fact that PSE meat syndrome occurs because of higher protein denaturation as the consequence of a simultaneous lower pH values under warm carcass sooner after slaughtering impairing the myofibril proteins functional properties. Conversely, PFN samples follow normal glycolysis rate maintaining the normal proteins activities. In conclusion, the results reported herein confirm the existence of this emerging broiler meat group with similar properties as control group and it should be considered as normal breast meat group.

Keywords: broiler breast meat, funcional properties, PFN, PSE

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Authors: Rita Bagheri, Javed Ahmed, Humayra Bashir, M. Irfan Qureshi

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Agriculture lands suffer from a combination of stresses such as salinity and metal contamination including cadmium at the same time. Under such condition of multiple stresses, plant may exhibit unique responses different from the stress occurring individually. Thus, it would be interesting to investigate that how plant respond to combined stress at level of antioxidants and proteome expression, and identifying the proteins which are involved in imparting stress tolerance. With an approach of comparative proteomics and antioxidant analysis, present study investigates the response of Spinacia oleracea to salt (NaCl), cadmium (Cd), and their combination (NaCl+Cd) stress. Two-dimensional gel electrophoresis was used for resolving leaf proteome, and proteins of interest were identified using PDQuest software. A number of proteins expressed differentially, those indicated towards their roles in imparting stress tolerance, were digested by trypsin and analyzed on mass spectrometer for peptide mass fingerprinting (PMF). Data signals were then matched with protein databases using MASCOT. Results show that NaCl, Cd and both together (NaCl+Cd) induce oxidative stress which was highest in combined stress of Cd+NaCl. Correspondingly, the activities of enzymatic antioxidants viz., SOD, APX, GR and CAT, and non-enzymatic antioxidants had highest changes under combined stress compares to single stress over their respective controls. Among the identified proteins, several interesting proteins were identified that may be have role in Spinacia oleracia tolerance in individual and combinatorial stress of salt and cadmium. The functional classification of identified proteins indicates the importance and necessity of keeping higher ratio of defence and disease responsive proteins.

Keywords: Spinacia oleracea, Cd, salinity, proteomics, antioxidants, combinatorial stress

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Authors: Vivek Kumar Gupta, Kripi Vohra, Monika Gupta

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Pulses have been a vital ingredient of the balanced human diet in India. Lentil (Lens culinaris Medikus or Lens esculenta Moench.) is a common legume known since biblical times. Lentil seeds, with or without hulls, are cooked as dhal and this has been the main dish for millennia in the South Asian region. Oxidative stress can damage lipids, proteins, enzymes, carbohydrates and DNA in cells and tissues, resulting in membrane damage, fragmentation or random cross linking of molecules like DNA, enzymes and structural proteins and even lead to cell death induced by DNA fragmentation and lipid peroxidation. These consequences of oxidative stress construct the molecular basis in the development of cancer, neurodegenerative disorders, cardiovascular diseases, diabetes and autoimmune. The aim of the present work is to assess the antioxidant potential of the peteroleum ether, acetone, methanol and water extract of the Lens esculenta seeds. In vitro antioxidant assessment of the extracts was carried out using 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging activity, hydroxyl radical scavenging activity, reducing power assay. The quantitative estimation of total phenolic content, total flavonoid content in extracts and in plant material, total saponin content, total alkaloid content, crude fibre content, total volatile content, fat content and mucilage content in drug material was also carried out. Though all the extracts exhibited dose dependent reducing power activity the acetone extract was found to possess significant hydrogen donating ability in DPPH (45.83%-93.13%) and hydroxyl radical scavenging system (28.7%-46.41%) than the peteroleum ether, methanol and water extracts. Total phenolic content in the acetone and methanol extract was found to be 608 and 188 mg gallic acid equivalent of phenol/g of sample respectively. Total flavonoid content of acetone and methanol extract was found to be 128 and 30.6 mg quercetin equivalent/g of sample respectively. It is evident that acetone extract of Lentil seeds possess high levels of polyphenolics and flavonoids that could be utilized as antioxidants and neutraceuticals.

Keywords: antioxidant, flavanoids, Lens esculenta, polyphenols

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Authors: Sara Przetocka, Krzysztof Żak, Grzegorz Dubin, Tadeusz Holak

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Toll-like receptors (TLRs) play central role in the innate immune response and inflammation by recognizing pathogen-associated molecular patterns (PAMPs). A fundamental basis of TLR signalling is dependent upon the recruitment and association of adaptor molecules that contain the structurally conserved Toll/interleukin-1 receptor (TIR) domain. MyD88 (myeloid differentiation primary response gene 88) is the universal adaptor for TLRs and cooperates with Mal (MyD88 adapter-like protein, also known as TIRAP) in TLR4 response which is predominantly used in inflammation, host defence and carcinogenesis. Up to date two possible models of MyD88, Mal and TLR4 interactions have been proposed. The aim of our studies is to confirm or abolish presented models and accomplish the full structural characterisation of TIR domains interaction. Using molecular cloning methods we obtained several construct of MyD88 and Mal TIR domain with GST or 6xHis tag. Gel filtration method as well as pull-down analysis confirmed that recombinant TIR domains from MyD88 and Mal are binding in complexes. To examine whether obtained complexes are homo- or heterodimers we carried out cross-linking reaction of TIR domains with BS3 compound combined with mass spectrometry. To investigate which amino acid residues are involved in this interaction the NMR titration experiments were performed. 15N MyD88-TIR solution was complemented with non-labelled Mal-TIR. The results undoubtedly indicate that MyD88-TIR interact with Mal-TIR. Moreover 2D spectra demonstrated that simultaneously Mal-TIR self-dimerization occurs which is necessary to create proper scaffold for Mal-TIR and MyD88-TIR interaction. Final step of this study will be crystallization of MyD88 and Mal TIR domains complex. This crystal structure and characterisation of its interface will have an impact in understanding the TLR signalling pathway and possibly will be used in development of new anti-cancer treatment.

Keywords: cancer, MyD88, TIR domains, Toll-like receptors

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Authors: Ume Habiba, Shafaqat Ali, Mujahid Farid, Muhammad Bilal Shakoor

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Copper (Cu) is an essential micronutrient for normal plant growth and development, but in excess, it is also toxic to plants. The present study investigated the influence of ethylenediaminetetraacetic acid (EDTA) in enhancing Cu uptake and tolerance as well as the morphological and physiological responses of Brassica napus L. seedlings under Cu stress. Four-week-old seedlings were transferred to hydroponics containing Hoagland’s nutrient solution. After 2 weeks of transplanting, three levels (0, 50, and 100 μM) of Cu were applied with or without application of 2.5 mM EDTA and plants were further grown for 8 weeks in culture media. Results showed that Cu alone significantly decreased plant growth, biomass, photosynthetic pigments, and gas exchange characteristics. Cu stress also reduced the activities of antioxidants, such as superoxide dismutase (SOD), peroxidase (POD), ascorbate peroxidase (APX), and catalase (CAT) along with protein contents. Cu toxicity increased the concentration of reactive oxygen species (ROS) as indicated by the increased production of malondialdehyde (MDA) and hydrogen peroxide (H2O2) in both leaves and roots. The application of EDTA significantly alleviated Cu-induced toxic effects in B. napus, showing remarkable improvement in all these parameters. EDTA amendment increased the activity of antioxidant enzymes by decreasing the concentrations of MDA and H2O2 both in leaves and roots of B. napus. Although, EDTA amendment with Cu significantly increased Cu uptake in roots, stems, and leaves in decreasing order of concentration but increased the growth, photosynthetic parameters, and antioxidant enzymes. These results showed that the application of EDTA can be a useful strategy for phytoextraction of Cu by B. napus from contaminated soils.

Keywords: antioxidants, biomass, copper, EDTA, phytoextraction, tolerance

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Authors: Said Belaaouad

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This study aimed to explore the quantitative structure-activity relationship (QSAR) of 1,2,4-Triazine-3(2H)-one derivative as a potential anticancer agent against breast cancer. The electronic descriptors were obtained using the Density Functional Theory (DFT) method, and a multiple linear regression techniques was employed to construct the QSAR model. The model exhibited favorable statistical parameters, including R2=0.849, R2adj=0.656, MSE=0.056, R2test=0.710, and Q2cv=0.542, indicating its reliability. Among the descriptors analyzed, absolute electronegativity (χ), total energy (TE), number of hydrogen bond donors (NHD), water solubility (LogS), and shape coefficient (I) were identified as influential factors. Furthermore, leveraging the validated QSAR model, new derivatives of 1,2,4-Triazine-3(2H)-one were designed, and their activity and pharmacokinetic properties were estimated. Subsequently, molecular docking (MD) and molecular dynamics (MD) simulations were employed to assess the binding affinity of the designed molecules. The Tubulin colchicine binding site, which plays a crucial role in cancer treatment, was chosen as the target protein. Through the simulation trajectory spanning 100 ns, the binding affinity was calculated using the MMPBSA script. As a result, fourteen novel Tubulin-colchicine inhibitors with promising pharmacokinetic characteristics were identified. Overall, this study provides valuable insights into the QSAR of 1,2,4-Triazine-3(2H)-one derivative as potential anticancer agent, along with the design of new compounds and their assessment through molecular docking and dynamics simulations targeting the Tubulin-colchicine binding site.

Keywords: QSAR, molecular docking, ADMET, 1, 2, 4-triazin-3(2H)-ones, breast cancer, anticancer, molecular dynamic simulations, MMPBSA calculation

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Authors: Kamyar Moradi

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Background: Acute mesenteric ischemia is known as a life-threatening condition. Re-establishment of blood flow in this condition can lead to mesenteric ischemia reperfusion (MIR) injury, which is accompanied by inflammatory response. Still, clear blueprint of inflammatory mechanism underlying MIR injury has not been provided. Interestingly, Albendazole has exhibited notable effects on inflammation and cytokine production. In this study, we aimed to evaluate outcomes of MIR injury following pretreatment with Albendazole with respect to assessment of mesenteric inflammation and ischemia threshold. Methods: Male rats were randomly divided into sham operated, vehicle treated, Albendazole 100 mg/kg, and Albendazole 200 mg/kg groups. MIR injury was induced by occlusion of superior mesenteric artery for 30 minutes followed by 120 minutes of reperfusion. Samples were utilized for assessment of epithelial survival and villous height. Immunohistochemistry study revealed intestinal expression of TNF-α and HIF-1-α. Gene expression of NF-κB/TLR4/TNF-α/IL-6 was measured using RTPCR. Also, protein levels of inflammatory cytokines in serum and intestine were assessed by ELISA method. Results: Histopathological study demonstrated that pretreatment with Albendazole could ameliorate decline in villous height and epithelial survival following MIR injury. Also, systemic inflammation was suppressed after administration of Albendazole. Analysis of possible participating inflammatory pathway could demonstrate that intestinal expression of NF-κB/TLR4/TNF-α/IL-6 is significantly attenuated in treated groups. Eventually, IHC study illustrated concordant decline in mesenteric expression of HIF-1-α/TNF-α. Conclusion: Single dose pretreatment with Albendazole could ameliorate inflammatory response and enhance ischemia threshold following induction of MIR injury. Still, more studies would clarify existing causality in this phenomenon.

Keywords: albendazole, ischemia reperfusion injury, inflammation, mesenteric ischemia

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Authors: Khushbu Priyadarshi, Chandramani Pathak

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Cancer cells can develop resistance to conventional therapies especially chemotherapeutic drugs. Resistance to chemotherapy is another challenge in cancer therapeutics. Therefore, it is important to address this issue. Gallic acid (GA) is a natural plant compound that exhibits various biological properties including anti-proliferative, anti-inflammatory, anti-oxidant and anti-bacterial. Despite of the wide spectrum biological properties GA has cytotoxic response and low bioavailability. To overcome this problem, GA was conjugated with the Polyamidoamine(PAMAM) dendrimer for improving the bioavailability and efficient delivery in drug-resistant HCT-116 Colon Cancer cells. Gallic acid was covalently linked to 4.0 G PAMAM dendrimer. PAMAM dendrimer is well established nanocarrier but has cytotoxicity due to presence of amphiphilic nature of amino group. In our study we have modified surface of PAMAM dendrimer with Gallic acid and examine their anti-proliferative effects in drug-resistant HCT-116 cells. Further, drug-resistant colon cancer cells were established and thereafter treated with different concentration of PAMAM-GA to examine their anti-proliferative potential. Our results show that PAMAM-GA conjugate induces apoptotic cell death in HCT-116 and drug-resistant cells observed by Annexin-PI staining. In addition, it also shows that multidrug-resistant drug transporter P-gp protein expression was downregulated with increasing the concentration of GA conjugate. After that we also observed the significant difference in Rh123 efflux and accumulation in drug sensitive and drug-resistant cancer cells. Thus, our study suggests that conjugation of anti-cancer agents with PAMAM could improve drug resistant property and cytotoxic response to treatment of cancer.

Keywords: drug resistance, gallic acid, PAMAM dendrimer, P-glycoprotein

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Authors: Cher Rong Matthew Kong, Edmund Tian, Seng Poon Ong, Chee Sian Gan

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Gamma-aminobutyric acid (GABA) is a non-protein amino acid that is a functional constituent of certain rice varieties. When consumed, it decreases blood pressure and reduces the risk of hypertension-related diseases. This has led to more research dedicated towards the development of functional food products (e.g. germinated brown rice) with enhanced GABA content, and the development of these functional food products has led to increased demand for instrument-based methods that can efficiently and effectively determine GABA content. Current analytical methods require analyte derivatisation, and have significant disadvantages such as being labour intensive and time-consuming, and being subject to analyte loss due to the increased complexity of the sample preparation process. To address this, an LC-MS-MS method for the determination of GABA in rice products has been developed and validated. This developed method involves a relatively simple sample preparation process before analysis using HILIC LC-MS-MS. This method eliminates the need for derivatisation, thereby significantly reducing the labour and time associated with such an analysis. Using LC-MS-MS also allows for better differentiation of GABA from any potential co-eluting compounds in the sample matrix. Results obtained from the developed method demonstrated high linearity, accuracy, and precision for the determination of GABA (1ng/L to 8ng/L) in a variety of brown rice products. The method can significantly simplify sample preparation steps, improve the accuracy of quantitation, and increase the throughput of analyses, thereby providing a quick but effective alternative to established instrumental analysis methods for GABA in rice.

Keywords: functional food, gamma-aminobutyric acid, germinated brown rice, method development

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Authors: Rostam Abdi, Ahmad Abdi, Zahra Vahedi Langrodi

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The aim of this study is to investigate four-week resistance exercise and milk supplement on NT-proBNP and plasma troponin I of male students. Concerning the methodology of the study, 21 senior high school students of Ardebil city were selected. The selected subjects were randomly shared in three groups of control, exercise- water and exercise- milk. The exercise program includes resistance exercise for a big muscle group. The subjects of control group rested during the study and did not participate in any training. The subjects of exercise- water experimental group immediately received 400 cc water after exercise and exercise- milk group immediately received 400 cc low fat milk. Control-water groups consumed the same amount of water. 48 hours before and after the last exercise session, the blood sample of the subjects were taken for measuring the variables. NT-proBNP and Troponin I concentrations were measured by ELISA. For data analysis, one-way variance analysis test, correlated t-test and Bonferroni post hoc test were used. The significant difference of p ≤ 0.05 was accepted. Resistance training along with milk consumption leads to increase of plasma NT-proBNP, however; this increase has not reached the significant level. Furthermore, meaningful increase was observed in plasma NT–proBNP in exercise group between pretest and posttest values. Furthermore, no meaningful difference was observed between groups in terms of Troponin I after milk consumption. It seems that endurance exercises lead to change in the structure of heart muscle and is along with an increase of NT-proBNP. Furthermore, there is the possibility that milk consumption can lead to release of heart troponin I. The mechanism through which protein supplements have been put on heart troponin I is unknown and requires more research.

Keywords: resistance exercise, milk, NT-proBNP, Troponin I

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