Search results for: protein modification

Authors: Richa Dhingra, Monika, Manav Malhotra, Tilak Raj Bhardwaj, Neelima Dhingra

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5-Alpha-reductases (5AR), a membrane bound, NADPH dependent enzyme and convert male hormone testosterone (T) into more potent androgen dihydrotestosterone (DHT). DHT is the required for the development and function of male sex organs, but its overproduction has been found to be associated with physiological conditions like Benign Prostatic Hyperplasia (BPH). Thus the inhibition of 5ARs could be a key target for the treatment of BPH. In present study, 2D and 3D Quantitative Structure Activity Relationship (QSAR) pharmacophore models have been generated for 5AR based on known inhibitory concentration (IC₅₀) values with extensive validations. The four featured 2D pharmacophore based PLS model correlated the topological interactions (–OH group connected with one single bond) (SsOHE-index); semi-empirical (Quadrupole2) and physicochemical descriptors (Mol. wt, Bromines Count, Chlorines Count) with 5AR inhibitory activity, and has the highest correlation coefficient (r² = 0.98, q² =0.84; F = 57.87, pred r² = 0.88). Internal and external validation was carried out using test and proposed set of compounds. The contribution plot of electrostatic field effects and steric interactions generated by 3D-QSAR showed interesting results in terms of internal and external predictability. The well validated 2D Partial Least Squares (PLS) and 3D k-nearest neighbour (kNN) models were used to search novel 5AR inhibitors with different chemical scaffold. To gain more insights into the molecular mechanism of action of these steroidal derivatives, molecular docking and in silico absorption, distribution, metabolism, and excretion (ADME) studies were also performed. Studies have revealed the hydrophobic and hydrogen bonding of the ligand with residues Alanine (ALA) 63A, Threonine (THR) 60A, and Arginine (ARG) 456A of 4AT0 protein at the hinge region. The results of QSAR, molecular docking, in silico ADME studies provide guideline and mechanistic scope for the identification of more potent 5-Alpha-reductase inhibitors (5ARI).

Keywords: 5α-reductase inhibitor, benign prostatic hyperplasia, ligands, molecular docking, QSAR

Procedia PDF Downloads 139

Authors: Cangir Uyarlar, Eyup Eren Gultepe, Mustafa Kabu, Hacı Ahmet Celik

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This study aimed to evaluate the effects of orally Immunoglobulin (Ig) Y treatments to calves were fed with colostrum having insufficient antibodies before first suckling. A total of 28 Holstein calves were fed assigned into control and treatment groups. The calves were fed fresh colostrum from their respective mother for the first 4 days. The treatment group calves were orally administered IgLock (10 g/d/calf) immediately before the first colostrum feeding and IgLock was administered just one time in treatment group calves. Then, the calves were offered normal milk until weaning. After weaning, all calves kept same paddock and were fed same ration. Diarrhea and respiratoric diseases were recorded for one year. Blood was collected from all calves in the study on birth day (0 day) before vaccination and IgLock administration, then, collected for the following 2 days in all groups. Albumin (ALB), Total Protein (TP), Aspartate Aminotransferase (AST), Alanine Aminotrasferase (ALT), Gamma-Glutamyl Transferase (GGT), Serum Amyloid A (SAA), Haptoglobin (HPT) and Ig G analyses were performed on all samples. Although serum ALB, ALT, GGT and Ig G levels were not shown a time dependent-change within control group; serum TP, AST, HPT and SAA levels were significantly changed by the time within mentioned group. Serum TP level was steady at first 2 days, then, it was increased significantly at 3rd day. Also, serum AST level was significantly increased at 2nd day, then it was descended to first day levels again at 3rd day. Although serum HPT levels were shown a significant gradually decreasing within control group, serum SAA levels were decreased rapidly after first day and there were no significance differences between 2nd and 3rd day in SAA levels. Serum ALB, ALT, HPT and SAA levels were not shown a time dependent-change within treatmet group. After first day Serum TP, GGT, AST and Ig G levels were shown an significant increasing at 2nd day. Serum TP, GGT and Ig G levels were higher as compared to 1st day within treatment group at 3rd day. But, serum AST level was less significantly 3rd day as compared to 2nd day values. The total numbers of calves suffered from diarrhea were significantly less in treatment group as compared to control group (p < 0,05). The pneumonia reappear ratio in calves suffered the diseases is 33,3% in control group and 11,11% in treatment group. Total cost of diseases and additives was 2339,36 $ for control and 1276,4 $ for treatment. As a conclusion, the immunity enhancers like IgLock are important and cost-effective to boost up immunity status in the early age which would be having positive effects on calves were received colostrum included insufficient antibodies.

Keywords: dairy calves, Ig Y, pneumonia, scours

Procedia PDF Downloads 474

Authors: S. Ibrahim, D. Salilew-Wondim, M. Hoelker, C. Looft, E. Tholen, C. Grosse-Brinkhaus, K. Schellander, C. Neuhoff, D. Tesfaye

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Bovine endometrial cells appear to have a key role in innate immune defense of the female genital tract. A better understanding of molecular changes in microRNAs (miRNAs) and their target genes expression may identify reliable prognostic indicators for cows that will resolve inflammation and resume cyclicity. In the current study, we hypothesized that let-7 miRNAs family has a primary role in the innate immune defence of the endometrium tissue against bacterial infection, which is partly achieved via regulating mRNA stability of pro-inflammatory cytokines at the post-transcriptional level. Therefore, we conducted two experiments. In the first experiment, primary bovine endometrial cells were challenged with clinical (3.0 μg/ml) and sub-clinical (0.5 μg/ml) doses of lipopolysaccharide (LPS) for 24h. In the 2nd experiment, we have investigated the potential role of let-7 miRNAs (let-7a and let-7f) using gain and loss of function approaches. Additionally, tumor necrosis factor alpha (TNFα), transforming growth factor beta 1 induced transcript 1 (TGFB1I1) and serum deprivation response (SDPR) genes were validated using reporter assay. Here we addressed for the first time that let-7 miRNAs have a precise role in bovine endometrium, where LPS dysregulated let-7 miRNAs family expression was associated with an increased pro-inflammatory cytokine level by directly/indirectly targeting the TNFα, interleukin 6 (IL6), nuclear factor kappa-light-chain enhancer of activated B cells (NFκB), TGFβ1I1 and SDPR genes. To our knowledge, this is the first study showing that TNFα, TGFβ1I1 and SDPR were identified and validated as novel let-7 miRNAs targets and could have a distinct role in inflammatory immune response of LPS challenged bovine endometrial cells. Our data represent a new finding by which uterine homeostasis is maintained through functional regulation of let-7a by down-regulation of pro-inflammatory cytokines expression (TNFα and IL6) at the mRNA and protein levels. These findings suggest that LPS serves as a negative regulator of let-7 miRNAs expression and provides a mechanism for the persistent pro-inflammatory phenotype, which is a hallmark of bovine subclinical endometritis.

Keywords: bovine endometrial cells, let-7, lipopolysaccharide, pro-inflammatory cytokines

Procedia PDF Downloads 351

Authors: Ahlam Said Al Azkawi, Nallusamy Sivakumar, Saif Nasser Al Bahri

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Diverse approaches and pathways are under development with the determination to develop cellulosic biofuels and other bio-products eventually at commercial scale in “bio-refineries”; however, the key challenge is mainly the high level of complexity in processing the feedstock which is complicated and energy consuming. To overcome the complications in utilizing the naturally occurring lignocellulose biomass, using waste paper as a feedstock for bio-production may solve the problem. Besides being abundant and cheap, bioprocessing of waste paper has evolved in response to the public concern from rising landfill cost from shrinking landfill capacity. Cardboard (CB) is one of the major components of municipal solid waste and one of the most important items to recycle. Although 50-70% of cardboard constitute is known to be cellulose and hemicellulose, the presence of lignin around them cause hydrophobic cross-link which physically obstructs the hydrolysis by rendering it resistant to enzymatic cleavage. Therefore, pretreatment is required to disrupt this resistance and to enhance the exposure of the targeted carbohydrates to the hydrolytic enzymes. Several pretreatment approaches have been explored, and the best ones would be those can influence cellulose conversion rates and hydrolytic enzyme performance with minimal or less cost and downstream processes. One of the promising strategies in this field is the application of surfactants, especially non-ionic surfactants. In this study, triton-X 100 was used as surfactants to treat cardboard prior enzymatic hydrolysis and compare it with acid treatment using 0.1% H2SO4. The effect of the surfactant enhancement was evaluated through its effect on hydrolysis rate in respect to time in addition to evaluating the structural changes and modification by scanning electron microscope (SEM) and X-ray diffraction (XRD) and through compositional analysis. Further work was performed to produce ethanol from CB treated with triton-X 100 via separate hydrolysis and fermentation (SHF) and simultaneous saccharification and fermentation (SSF). The hydrolysis studies have demonstrated enhancement in saccharification by 35%. After 72 h of hydrolysis, a saccharification rate of 98% was achieved from CB enhanced with triton-X 100, while only 89 of saccharification achieved from acid pre-treated CB. At 120 h, the saccharification % exceeded 100 as reducing sugars continued to increase with time. This enhancement was not supported by any significant changes in the cardboard content as the cellulose, hemicellulose and lignin content remained same after treatment, but obvious structural changes were observed through SEM images. The cellulose fibers were clearly exposed with very less debris and deposits compared to cardboard without triton-X 100. The XRD pattern has also revealed the ability of the surfactant in removing calcium carbonate, a filler found in waste paper known to have negative effect on enzymatic hydrolysis. The cellulose crystallinity without surfactant was 73.18% and reduced to 66.68% rendering it more amorphous and susceptible to enzymatic attack. Triton-X 100 has proved to effectively enhance CB hydrolysis and eventually had positive effect on the ethanol yield via SSF. Treating cardboard with only triton-X 100 was a sufficient treatment to enhance the enzymatic hydrolysis and ethanol production.

Keywords: cardboard, enhancement, ethanol, hydrolysis, treatment, Triton-X 100

Procedia PDF Downloads 122

Authors: Ribeiro Tiago Magalhães

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Objective: The objective of this study was to evaluate the influence of a hydroelectric power plant in the state of Amapá, and to present the results obtained by dimensioning the diversity of the main mosquito vectors involved in the transmission of pathogens that cause diseases such as malaria, dengue and leishmaniasis. Methodology: The present study was conducted on the banks of the Araguari River, in the municipalities of Porto Grande and Ferreira Gomes in the southern region of Amapá State. Nine monitoring campaigns were conducted, the first in April 2014 and the last in March 2016. The selection of the catch sites was done in order to prioritize areas with possible occurrence of the species considered of greater importance for public health and areas of contact between the wild environment and humans. Sampling efforts aimed to identify the local vector fauna and to relate it to the transmission of diseases. In this way, three phases of collection were established, covering the schedules of greater hematophageal activity. Sampling was carried out using Shannon Shack and CDC types of light traps and by means of specimen collection with the hold method. This procedure was carried out during the morning (between 08:00 and 11:00), afternoon-twilight (between 15:30 and 18:30) and night (between 18:30 and 22:00). In the specific methodology of capture with the use of the CDC equipment, the delimited times were from 18:00 until 06:00 the following day. Results: A total of 32 species of mosquitoes was identified, and a total of 2,962 specimens was taxonomically subdivided into three genera (Culicidae, Psychodidae and Simuliidae) Psorophora, Sabethes, Simulium, Uranotaenia and Wyeomyia), besides those represented by the family Psychodidae that due to the morphological complexities, allows the safe identification (without the method of diaphanization and assembly of slides for microscopy), only at the taxonomic level of subfamily (Phlebotominae). Conclusion: The nine monitoring campaigns carried out provided the basis for the design of the possible epidemiological structure in the areas of influence of the Cachoeira Caldeirão HPP, in order to point out among the points established for sampling, which would represent greater possibilities, according to the group of identified mosquitoes, of disease acquisition. However, what should be mainly considered, are the future events arising from reservoir filling. This argument is based on the fact that the reproductive success of Culicidae is intrinsically related to the aquatic environment for the development of its larvae until adulthood. From the moment that the water mirror is expanded in new environments for the formation of the reservoir, a modification in the process of development and hatching of the eggs deposited in the substrate can occur, causing a sudden explosion in the abundance of some genera, in special Anopheles, which holds preferences for denser forest environments, close to the water portions.

Keywords: Amazon, hydroelectric, power, plants

Procedia PDF Downloads 157

Authors: Abayomi O. Adetuyi, Jamiu M. Jabar, Samuel O. Afolabi

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Vegetable fibres are classes of fibres of low density, biodegradable and non-abrasive that are largely abundant fibre materials with specific properties and mostly found/ obtained in plants on earth surface. They are classified into three categories, depending on the part of the plant from which they are gotten from namely: fruit, Blast and Leaf fibre. Ever since four/five millennium B.C, attention has been focussing on the commonest and highly utilized cotton fibre obtained from the fruit of cotton plants (Gossypium spp), for the production of cotton fabric used in every home today. The present study, therefore, focused on the ability of three underutilized vegetable (fruit) fibres namely: coir fiber (Eleas coniferus), palm kernel fiber and empty fruit bunch fiber (Elias guinensis) through chemical modifications for better composite value addition performance to polyurethane form and dye adsorption. These fibres were sourced from their parents’ plants, identified and cleansed with 2% hot detergent solution 1:100, rinsed in distilled water and oven-dried to constant weight, before been chemically modified through alkali bleaching, mercerization and acetylation. The alkali bleaching involves treating 0.5g of each fiber material with 100 mL of 2% H2O2 in 25 % NaOH solution with refluxing for 2 h. While that of mercerization and acetylation involves the use of 5% sodium hydroxide NaOH solution for 2 h and 10% acetic acid- acetic anhydride 1:1 (v/v) (CH3COOH) / (CH3CO)2O solution with conc. H2SO4 as catalyst for 1 h, respectively on the fibres. All were subsequently washed thoroughly with distilled water and oven dried at 105 0C for 1 h. These modified fibres were incorporated as composite into polyurethane form and used in dye adsorption study of indigo. The first two treatments led to fiber weight reduction, while the acidified acetic anhydride treatment gave the fibers weight increment. All the treated fibers were found to be of less hydrophilic nature, better mechanical properties, higher thermal stabilities as well as better adsorption surfaces/capacities than the untreated ones. These were confirmed by gravimetric analysis, Instron Universal Testing Machine, Thermogravimetric Analyser and the Scanning Electron Microscope (SEM) respectively. The fiber morphology of the modified fibers showed smoother surfaces than unmodified fibres.The empty fruit bunch fibre and the coconut coir fibre are better than the palm kernel fibres as reinforcers for composites or as adsorbents for waste-water treatment. Acetylation and alkaline bleaching treatment improve the potentials of the fibres more than mercerization treatment. Conclusively, vegetable fibres, especially empty fruit bunch fibre and the coconut coir fibre, which are cheap, abundant and underutilized, can replace the very costly powdered activated carbon in wastewater treatment and as reinforcer in foam.

Keywords: chemical modification, industrial application, value addition, vegetable fibre

Procedia PDF Downloads 297

Authors: Fakunle Egbo, Kammalchukwu A., Akinremi T.

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Physical activity and nutritional status influence the health status and cognition of young adults. Lack of physical activity increases the likelihood of developing obesity which leads to the risk of heart diseases and other risk factors like high blood pressure, high blood cholesterol, diabetes etc. The study employed a cross-sectional study design. The study used a multi stage sampling technique multi- stage sampling technique; Purposive, for the selection of colleges that would be used, stratified random sampling for stratifying the colleges into departments and the simple random sampling for the selection of each respondent from the departments. Structured questionnaires were used to obtain data from the respondents and pre-tested anthropometric instruments were used to get the weight and height of the respondents and statistically analyzed using SPSS version 22.0 and the TDA (Total dietary allowance) software which was used to analyze the nutrient intake of the respondents. This study showed that they comprised of 50.1% males and 40.9% females. Slightly above average 51.8% were between ages of 15-19 with mean age being 19.57 years; ages 20-24 were slightly below average at 45.7%. The male students 58.7% had vigorous physical activity, whereas majority of females 76.5% had light physical activity level. 39.1% of the male students carried out physical activity 2-3 times per week while One third of the female students (38.3%) carried out physical activity 6-7 times per week. Majority of the respondents had Inadequate Protein- 63.8%, Carbohydrate- 60.2%, and Dietary fiber- 88.8. 36% eat rice 4-6 times per week. Majority of the respondents had inadequate fruit and vegetables (Efo, Banana,) at 47.7%, 40.6% respectively. Using Body mass index, (63.2%) have normal weight. 22.9% are overweight, 6.8% are underweight, 5.4% have grade 1 obesity and 1.6% have grade II obesity. There was a statistically significant association between the physical activity of the respondents with their nutritional status (p=0.037), physical activity and sex (p=0.000), nutritional status and amount spent on food daily (p=0.007). The study concluded that the physical activity level of the respondents, most especially the females were low; One third of the students were malnourished therefore, there should be an urgent need for improving the overall health status of students by providing the students with well-equipped gyms and other sporting equipment’s that would make them participate actively and keep fit.

Keywords: physical activity, nutritional status, undergraduates, dietary pattern

Procedia PDF Downloads 45

Authors: Oqba Basal, András Szabó

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Legume crops are able to fix atmospheric nitrogen by the symbiotic relation with specific bacteria, which allows the use of the mineral nitrogen-fertilizer to be reduced, or even excluded, resulting in more profit for the farmers and less pollution for the environment. Soybean (Glycine max (L.) Merrill) is one of the most important legumes with its high content of both protein and oil. However, it is recommended to combine the two nitrogen sources under stress conditions in order to overcome its negative effects. Drought stress is one of the most important abiotic stresses that increasingly limits soybean yields. A precise rate of mineral nitrogen under drought conditions is not confirmed, as it depends on many factors; soybean yield-potential and soil-nitrogen content to name a few. An experiment was conducted during 2017 growing season in Debrecen, Hungary to investigate the effects of nitrogen source on the physiology and the yield of the soybean cultivar 'Boglár'. Three N-fertilizer rates including no N-fertilizer (0 N), 35 kg ha^-1 of N-fertilizer (35 N) and 105 kg ha^-1 of N-fertilizer (105 N) were applied under three different irrigation regimes; severe drought stress (SD), moderate drought stress (MD) and control with no drought stress (ND). Half of the seeds in each treatment were pre-inoculated with Bradyrhizobium japonicum inoculant. The overall results showed significant differences associated with fertilization and irrigation, but not with inoculation. Increasing N rate was mostly accompanied with increased chlorophyll content and leaf area index, whereas it positively affected the plant height only when the drought was waived off. Plant height was the lowest under severe drought, regardless of inoculation and N-fertilizer application and rate. Inoculation increased the yield when there was no drought, and a low rate of N-fertilizer increased the yield furthermore; however, the high rate of N-fertilizer decreased the yield to a level even less than the inoculated control. On the other hand, the yield of non-inoculated plants increased as the N-fertilizer rate increased. Under drought conditions, adding N-fertilizer increased the yield of the non-inoculated plants compared to their inoculated counterparts; moreover, the high rate of N-fertilizer resulted in the best yield. Regardless of inoculation, the mean yield of the three fertilization rates was better when the water amount increased. It was concluded that applying N-fertilizer to provide the nitrogen needed by soybean plants, with the absence of N₂-fixation process, is very important. Moreover, adding relatively high rate of N-fertilizer is very important under severe drought stress to alleviate the drought negative effects. Further research to recommend the best N-fertilizer rate to inoculated soybean under drought stress conditions should be executed.

Keywords: drought stress, inoculation, N-fertilizer, soybean physiology, yield

Procedia PDF Downloads 123

Authors: Saeed Qaisrani

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The study was conducted to observe carbon and nutrient loss by burning of rice residues on rice-wheat cropping system The rice crop was harvested to conduct the experiment in a randomized complete block design (RCBD) with factors and 4 replications with a net plot size of 10 m x 20 m. Rice stubbles were managed by two methods i.e. Incorporation & burning of rice residues. Soil samples were taken to a depth of 30 cm before sowing & after harvesting of wheat. Wheat was sown after harvesting of rice by three practices i.e. Conventional tillage, Minimum tillage and Zero tillage to observe best tillage practices. Laboratory and field experiments were conducted on wheat to assess best tillage practice and residues management method with estimation of carbon losses. Data on the following parameters; establishment count, plant height, spike length, number of grains per spike, biological yield, fat content, carbohydrate content, protein content, and harvest index were recorded to check wheat quality & ensuring food security in the region. Soil physico-chemical analysis i.e. pH, electrical conductivity, organic matter, nitrogen, phosphorus, potassium, and carbon were done in soil fertility laboratory. Substantial results were found on growth, yield and related parameters of wheat crop. The collected data were examined statistically with economic analysis to estimate the cost-benefit ratio of using different tillage techniques and residue management practices. Obtained results depicted that Zero tillage method have positive impacts on growth, yield and quality of wheat, Moreover, it is cost effective methodology. Similarly, Incorporation is suitable and beneficial method for soil due to more nutrients provision and reduce the need of fertilizers. Burning of rice stubbles has negative impact including air pollution, nutrient loss, microbes died and carbon loss. Recommended the zero tillage technology to reduce carbon losses along with food security in Pakistan.

Keywords: agricultural agronomy, food security, carbon sequestration, rice-wheat cropping system

Procedia PDF Downloads 255

Authors: Jyh-Ping Chen, Chia-Lin Sheu

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In this study, we propose to use electrospinning to fabricate porous nanofibrous membranes as postsurgical anti-adhesion barriers and to improve the properties of current post-surgical anti-adhesion products. We propose to combine FDA-approved biomaterials with anti-adhesion properties, polycaprolactone (PCL), polyethylene glycol (PEG), hyaluronic acid (HA) with silver nanoparticles (Ag) and ibuprofen (IBU), to produce anti-adhesion barrier nanofibrous membranes. For this purpose, PEG/PCL/Ag/HA/IBU core-shell nanofibers were prepared. The shell layer contains PEG + PCL to provide mechanical supports and Ag was added to the outer PEG-PCL shell layer during electrospinning to endow the nanofibrous membrane with anti-bacterial properties. The core contains HA to exert anti-adhesion and IBU to exert anti-inflammation effects, respectively. The nanofibrous structure of the membranes can reduce cell penetration while allowing nutrient and waste transports to prevent postsurgical adhesion. Nanofibers with different core/shell thickness ratio were prepared. The nanofibrous membranes were first characterized for their physico-chemical properties in detail, followed by in vitro cell culture studies for cell attachment and proliferation. The HA released from the core region showed extended release up to 21 days for prolonged anti-adhesion effects. The attachment of adhesion-forming fibroblasts is reduced using the nanofibrous membrane from DNA assays and confocal microscopic observation of adhesion protein vinculin expression. The Ag released from the shell showed burst release to prevent E Coli and S. aureus infection immediately and prevent bacterial resistance to Ag. Minimum cytotoxicity was observed from Ag and IBU when fibroblasts were culture with the extraction medium of the nanofibrous membranes. The peritendinous anti-adhesion model in rabbits and the peritoneal anti-adhesion model in rats were used to test the efficacy of the anti-adhesion barriers as determined by gross observation, histology, and biomechanical tests. Within all membranes, the PEG/PCL/Ag/HA/IBU core-shell nanofibers showed the best reduction in cell attachment and proliferation when tested with fibroblasts in vitro. The PEG/PCL/Ag/HA/IBU nanofibrous membranes also showed significant improvement in preventing both peritendinous and peritoneal adhesions when compared with other groups and a commercial adhesion barrier film.

Keywords: anti-adhesion, electrospinning, hyaluronic acid, ibuprofen, nanofibers

Procedia PDF Downloads 156

Authors: Xuechun Kan, Dongdong Li, Fan Li, Peidang Liu

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Triple-negative breast cancer (TNBC) is the most malignant subtype of breast cancer with a poor prognosis, and radiotherapy is one of the main treatment methods. However, due to the obvious resistance of tumor cells to radiotherapy, high dose of ionizing radiation is required during radiotherapy, which causes serious damage to normal tissues near the tumor. Therefore, how to improve radiotherapy resistance and enhance the specific killing of tumor cells by radiation is a hot issue that needs to be solved in clinic. Recent studies have shown that silver-based nanoparticles have strong radiosensitization, and silver nanoclusters (AgNCs) also provide a broad prospect for tumor targeted radiosensitization therapy due to their ultra-small size, low toxicity or non-toxicity, self-fluorescence and strong photostability. Aptamer 5TR1 is a 25-base oligonucleotide aptamer that can specifically bind to mucin-1 highly expressed on the membrane surface of TNBC 4T1 cells, and can be used as a highly efficient tumor targeting molecule. In this study, AgNCs were synthesized by DNA template based on 5TR1 aptamer (NC-T5-5TR1), and its role as a targeted radiosensitizer in TNBC radiotherapy was investigated. The optimal DNA template was first screened by fluorescence emission spectroscopy, and NC-T5-5TR1 was prepared. NC-T5-5TR1 was characterized by transmission electron microscopy, ultraviolet-visible spectroscopy and dynamic light scattering. The inhibitory effect of NC-T5-5TR1 on cell activity was evaluated using the MTT method. Laser confocal microscopy was employed to observe NC-T5-5TR1 targeting 4T1 cells and verify its self-fluorescence characteristics. The uptake of NC-T5-5TR1 by 4T1 cells was observed by dark-field imaging, and the uptake peak was evaluated by inductively coupled plasma mass spectrometry. The radiation sensitization effect of NC-T5-5TR1 was evaluated through cell cloning and in vivo anti-tumor experiments. Annexin V-FITC/PI double staining flow cytometry was utilized to detect the impact of nanomaterials combined with radiotherapy on apoptosis. The results demonstrated that the particle size of NC-T5-5TR1 is about 2 nm, and the UV-visible absorption spectrum detection verifies the successful construction of NC-T5-5TR1, and it shows good dispersion. NC-T5-5TR1 significantly inhibited the activity of 4T1 cells and effectively targeted and fluoresced within 4T1 cells. The uptake of NC-T5-5TR1 reached its peak at 3 h in the tumor area. Compared with AgNCs without aptamer modification, NC-T5-5TR1 exhibited superior radiation sensitization, and combined radiotherapy significantly inhibited the activity of 4T1 cells and tumor growth in 4T1-bearing mice. The apoptosis level of NC-T5-5TR1 combined with radiation was significantly increased. These findings provide important theoretical and experimental support for NC-T5-5TR1 as a radiation sensitizer for TNBC.

Keywords: 5TR1 aptamer, silver nanoclusters, radio sensitization, triple-negative breast cancer

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Authors: Sonia Kudlacik-Kramarczyk, Anna Drabczyk, Dagmara Malina, Bozena Tyliszczak, Agnieszka Sobczak-Kupiec

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Introduction: In the preparation of polymer materials, compounds of natural origin are currently gaining more and more interest. This is particularly noticeable in the case of synthesis of materials considered for biomedical use. Then, selected material has to meet many requirements. It should be characterized by non-toxicity, biodegradability and biocompatibility. Therefore special attention is directed to substances such as polysaccharides, proteins or substances that are the basic building components of proteins, i.e. amino acids. These compounds may be crosslinked with other reagents that leads to the preparation of polymer matrices. Such amino acids as e.g. cysteine or histidine. On the other hand, previously mentioned requirements may be met by polymers obtained as a result of biosynthesis, e.g. polyhydroxybutyrate. This polymer belongs to the group of aliphatic polyesters that is synthesized by microorganisms (selected strain of bacteria) under specific conditions. It is possible to modify matrices based on given polymer with substances of various origin. Such a modification may result in the change of their properties or/and in providing the material with new features desirable in viewpoint of specific application. Described materials are synthesized using UV radiation. Process of photopolymerization is fast, waste-free and enables to obtain final products with favorable properties. Methodology: Polymer matrices have been prepared by means of photopolymerization. First step involved the preparation of solutions of particular reagents and mixing them in the appropriate ratio. Next, crosslinking agent and photoinitiator have been added to the reaction mixture and the whole was poured into the Petri dish and treated with UV radiation. After the synthesis, polymer samples were dried at room temperature and subjected to the numerous analyses aimed at the determining their physicochemical properties. Firstly, sorption properties of obtained polymer matrices have been determined. Next, mechanical properties have been characterized, i.e. tensile strength. The ability to deformation under applied stress of all prepared polymer matrices has been checked. Such a property is important in viewpoint of the application of analyzed materials e.g. as wound dressings. Wound dressings have to be elastic because depending on the location of the wound and its mobility, such a dressing has to adhere properly to the wound. Furthermore, considering the use of the materials for biomedical purposes it is essential to determine its behavior in environments simulating these ones occurring in human body. Therefore incubation studies using selected liquids have also been conducted. Conclusions: As a result of photopolymerization process, polymer matrices based on natural compounds have been prepared. These exhibited favorable mechanical properties and swelling ability. Moreover, biocompatibility in relation to simulated body fluids has been stated. Therefore it can be concluded that analyzed polymer matrices constitute an interesting materials that may be considered for biomedical use and may be subjected to the further more advanced analyses using specific cell lines.

Keywords: photopolymerization, polymer matrices, simulated body fluids, swelling properties

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Authors: Sonia Bautista Carrascosa, Carlos Renedo Sanchez

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The effect of a rapid drawdown is a classical scenario to be considered in slope stability under submerged conditions. This situation arises when totally or partially submerged slopes experience a descent of the external water level and is a typical verification to be done in a dam engineering discipline, as reservoir water levels commonly fluctuate noticeably during seasons and due to operational reasons. Although the scenario is well known and predictable in general, site conditions can increase the complexity of its assessment and external factors are not always expected, can cause a reduction in the stability or even a failure in a slope under a rapid drawdown situation. The present paper describes and discusses the interaction between two different infrastructures, a dam and a highway, and the impact on the stability of a natural rock slope overlaid by the north abutment of a viaduct of the A-44 Highway due to the rapid drawdown of the Rules Dam, in the province of Granada (south of Spain). In the year 2011, with both infrastructures, the A-44 Highway and the Rules Dam already constructed, delivered and under operation, some movements start to be recorded in the approximation embankment and north abutment of the Guadalfeo Viaduct, included in the highway and developed to solve the crossing above the tail of the reservoir. The embankment and abutment were founded in a low-angle natural rock slope formed by grey graphic phyllites, distinctly weathered and intensely fractured, with pre-existing fault and weak planes. After the first filling of the reservoir, to a relative level of 243m, three consecutive drawdowns were recorded in the autumns 2010, 2011 and 2012, to relative levels of 234m, 232m and 225m. To understand the effect of these drawdowns in the weak rock mass strength and in its stability, a new geological model was developed, after reviewing all the available ground investigations, updating the geological mapping of the area and supplemented with an additional geotechnical and geophysical investigations survey. Together with all this information, rainfall and reservoir level evolution data have been reviewed in detail to incorporate into the monitoring interpretation. The analysis of the monitoring data and the new geological and geotechnical interpretation, supported by the use of limit equilibrium software Slide2, concludes that the movement follows the same direction as the schistosity of the phyllitic rock mass, coincident as well with the direction of the natural slope, indicating a deep-seated movement of the whole slope towards the reservoir. As part of these conclusions, the solutions considered to reinstate the highway infrastructure to the required FoS will be described, and the geomechanical characterization of these weak rocks discussed, together with the influence of water level variations, not only in the water pressure regime but in its geotechnical behavior, by the modification of the strength parameters and deformability.

Keywords: monitoring, rock slope stability, water drawdown, weak rock

Procedia PDF Downloads 138

Authors: I. Upadhyaya, K. Arsi, A. M. Donoghue, C. N. Coon, M. Schlumbohm, M. N. Riaz, M. B. Farnell, A. Upadhyay, A. J. Davis, D. J. Donoghue

Abstract:

Methionine, a sulfur containing amino acid, is essential for healthy poultry production. Synthetic methionine is commonly used as a supplement in conventional poultry. However, for organic poultry, a natural, cost effective source of methionine that can replace synthetic methionine is unavailable. Invasive Asian carp (AC) are a potential natural methionine source; however, there is no proven technology to utilize this fish methionine. Commercially available rendering is environmentally challenging due to the offensive smell produced during production. We explored extrusion technology as a potential cost effective alternative to fish rendering. We also determined the amino acid composition, digestible amino acids and total metabolizable energy (TMEn) for the extruded AC fish meal. Dry extrusion of AC was carried out by mixing the fish with soybean meal (SBM) in a 1:1 proportion to reduce high moisture in the fishmeal using an Insta Pro Jr. dry extruder followed by drying and grinding of the product. To determine the digestible amino acids and TMEn of the extruded product, a colony of cecectomized Bovans White Roosters was used. Adult roosters (48 weeks of age) were fasted for 30 h and tube fed 35 grams of 3 treatments: (1) extruded AC fish meal, (2) SBM and (3) corn. Excreta from each individual bird was collected for the next 48 h. An additional 10 unfed roosters served as endogenous controls. The gross energy and protein content of the feces from the treatments were determined to calculate the TMEn. Fecal samples and treatment feeds were analyzed for amino acid content and percent digestible amino acid. Results from the analysis suggested that addition of Asian carp increased the methionine content of SBM from 0.63 to 0.83%. Also, the digestibility of amino acid and the TMEn values were greater for the AC meal with SBM than SBM alone. The dry extruded AC meal analysis is indicative that the product can replace SBM alone and enhance natural methionine in a standard poultry ration. The results from feed formulation using different concentrations of the AC fish meal depict a potential diet which can supplement the required methionine content in organic poultry production.

Keywords: Asian carp, extrusion, natural methionine, organic poultry

Procedia PDF Downloads 192

Authors: Mahabub Alam, Mohammad Mahmudul Hassan, M. Hasanuzzaman, M. Ahasanul Hoque

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Animals are transported from one place to another for different purposes in Bangladesh. However, the potential effect of long-distance transport on cattle health has not frequently been studied. Therefore, this study was conducted to assess health conditions of cattle transported from a long distance to Chittagong in Bangladesh. A total of 100 adult cattle, regardless of breed and sex, were selected at Benapole live cattle market in Jessore between August and September 2015 for the study. Blood samples were taken from 50 randomly selected cattle at 0 hours before transportation, just after transportation, at 12-16 hours post-conclusion of transportation, and 24 hours after transportation. The external health conditions and injuries of the cattle were assessed by close inspection, and the trader was interviewed using the structured questionnaire. Images of cattle injuries were taken with a camera. The basic internal health of the cattle was evaluated using standard hemato-biochemical tests. Animals were fasted and remained standing within a small space allocation (8-10 sq feet/animal) in the vehicle during transportation. Animals were provided only with paddy straw and water prior to selling at the destination market. The overall frequency of cattle injuries varied significantly (26% before vs. 47% after transportation; p < 0.001). The frequency of different cattle injuries also significantly varied by types such as abrasion (11% vs. 21%; p < 0.05) and barbed wire injury (9% vs. 18%; p < 0.05). Single cattle injury differed significantly (21% vs. 36%; p < 0.001). Cattle health conditions varied significantly (nasal discharge: 15% vs. 28%; p < 0.05; diarrhea: 15% vs. 23%; p < 0.05 and severe dehydration: 8% vs. 20%; p < 0.001). The values of hemoglobin (Hb), total erythrocyte count (TEC), total leukocyte count (TLC), lymphocyte (L), neutrophil (N) and eosinophil (E) varied significantly (p ≤ 0.01) (Hb: 11.1mg/dl vs. 12.3mg/dl; TEC: 4.7 million/ml vs. 5.7million/ml; TLC: 6.2 thousand/ml vs. 7.3 thousand/ml; L: 61.7% vs. 58.1%; N: 29.7% vs. 32.8%; E: 3.8% vs. 4.7%). The values of serum total protein (TP), creatine kinase (CK), triglyceride (TG), calcium (Ca), phosphorus (P) and alkaline phosphatase (ALP) significantly differed (p ≤ 0.05) (TP: 6.8g/dl vs. 8.2g/dl; CK:574.9u/l vs. 1288u/l; TG: 104.7mg/dl vs. 127.7mg/dl; Ca: 11.3mg/dl vs. 13mg/dl; P: 7.3mg/dl vs. 7.6mg/dl; ALP: 303u/l vs. 363u/l). The identified status of external and internal health conditions of the cattle for trading purpose due to long-distance transportation in the present study indicates a high degree of transport stress and poor animal welfare.

Keywords: animal welfare, cattle, external and internal health conditions, transportation

Procedia PDF Downloads 153

Authors: Moustafa Osman Mohammed

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This paper introduces Luhmann’s autopoietic social systems starting with the original concept of autopoiesis by biologists and scientists, including the modification of general systems based on socialized medicine. A specific type of autopoietic system is explained in the three existing groups of the ecological phenomena: interaction, social and medical sciences. This hypothesis model, nevertheless, has a nonlinear interaction with its natural environment ‘interactional cycle’ for the exchange of photon energy with molecular without any changes in topology. The external forces in the systems environment might be concomitant with the natural fluctuations’ influence (e.g. radioactive radiation, electromagnetic waves). The cantilever sensor deploys insights to the future chip processor for prevention of social metabolic systems. Thus, the circuits with resonant electric and optical properties are prototyped on board as an intra–chip inter–chip transmission for producing electromagnetic energy approximately ranges from 1.7 mA at 3.3 V to service the detection in locomotion with the least significant power losses. Nowadays, therapeutic systems are assimilated materials from embryonic stem cells to aggregate multiple functions of the vessels nature de-cellular structure for replenishment. While, the interior actuators deploy base-pair complementarity of nucleotides for the symmetric arrangement in particular bacterial nanonetworks of the sequence cycle creating double-stranded DNA strings. The DNA strands must be sequenced, assembled, and decoded in order to reconstruct the original source reliably. The design of exterior actuators have the ability in sensing different variations in the corresponding patterns regarding beat-to-beat heart rate variability (HRV) for spatial autocorrelation of molecular communication, which consists of human electromagnetic, piezoelectric, electrostatic and electrothermal energy to monitor and transfer the dynamic changes of all the cantilevers simultaneously in real-time workspace with high precision. A prototype-enabled dynamic energy sensor has been investigated in the laboratory for inclusion of nanoscale devices in the architecture with a fuzzy logic control for detection of thermal and electrostatic changes with optoelectronic devices to interpret uncertainty associated with signal interference. Ultimately, the controversial aspect of molecular frictional properties is adjusted to each other and forms its unique spatial structure modules for providing the environment mutual contribution in the investigation of mass temperature changes due to pathogenic archival architecture of clusters.

Keywords: autopoiesis, nanoparticles, quantum photonics, portable energy, photonic structure, photodynamic therapeutic system

Procedia PDF Downloads 98

Authors: Engin Berber, Nurettin Canakoglu, Ibrahim Sozdutmaz, Merve Caliskan, Shaikh Terkis Islam Pavel, Hazel Yetiskin, Aykut Ozdarendeli

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Crimean-Congo hemorrhagic fever virus (CCHFV) is a tick-borne virus in the family Bunyaviridae, genus Nairovirus. The CCHFV genome consists of three molecules of negative-sense single-stranded RNA, each encapsulated separately. The virion particle contains viral RNA polymerase (L segment), surface glycoproteins Gn and Gc (Msegment), and a nucleocapsid protein NP (S segment). CCHF is characterized by high case mortality, occurring in Asia, Africa, the Middle East and Eastern Europe. Clinical CCHF was first recognized in Turkey in 2002. The numbers of CCHF cases have gradually increased in Turkey making the virus a public health concern. Between 2002 and 2014, more than 8000 the CCHF cases have been reported in Turkey and mortality rate is around 5%. So, Turkey is one of the countries where the epidemy has become spread to the wider geography and the biggest outbreaks of CCHF have occurred in the world. We have recently developed an inactivated cell-culture based vaccine against CCHF. We have showed that the Balb/c mice immunized with the CCHF vaccine induced the high level of neutralizing antibodies. In this study, we aimed to determine the immunodominant regions of nucleoprotein (NP) CCHFV Kelkit06 strain which stimulate T cells. For this purpose, pools of overlapping NP were used for an IFN- γ ELISPOT assay. Balb/c mice were divided into two groups for the experiment. Two groups (n = 10 each) were immunized via the intraperitoneal route with 5, or 10μg of the cell culture-based vaccine. The control group (n = 6) was mock immunized with PBS. Booster injections with the same formulation were given on days 21 and 42 after the first immunization. The higher reactivity against the CCHFV NP pools 31-40 and 80-90 was determined in the two dose groups. In order to analyze the vaccine-induced T cell responses in Balb/c mice immunized with varying doses of the vaccine, we have been also currently working on CD4+, CD8+ and CD3 + T cells by flow cytometry.

Keywords: Crimean-Congo hemorrhagic fever virus, immunodominant regions of NP, T cell response, vaccine

Procedia PDF Downloads 318

Authors: Mwafaq Ibdah, Mossab, Yahyaa, Dor Rachmany, Yoram Gerchman, Doron Holland, Liora Shaltiel-Harpaz

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Pear Psylla is the most important pest of pear in all pear-growing regions, in Asian, European, and the USA. Pear psylla damages pears in several ways: high-density populations of these insects can cause premature leaf and fruit drop, diminish plant growth, and reduce fruit size. In addition, their honeydew promotes sooty mold on leaves and russeting on fruit. Pear psyllas are also considered vectors of pear pathogens such as Candidatus Phytoplasma pyri causing pear decline that can lead to loss of crop and tree vigor, and sometimes loss of trees. Psylla control is a major obstacle to efficient integrated pest management. Recently we have identified two naturally resistance pear accessions (Py.760-261 and Py.701-202) in the Newe Ya’ar live collection. GC-MS volatile metabolic profiling identified several volatile compounds common in these accessions but lacking, or much less common, in a sensitive accession, the commercial Spadona variety. Among these volatiles were styrene and its derivatives. When the resistant accessions were used as inter-stock, the volatile compounds appear in commercial Spadona scion leaves, and it showed reduced susceptibility to pear psylla. Laboratory experiments and applications of some of these volatile compounds were very effective against psylla eggs, nymphs, and adults. The genes and enzymes involved in the specific reactions that lead to the biosynthesis of styrene in plant are unknown. We have identified a phenolic acid decarboxylase that catalyzes the formation of p-hydroxystyrene, which occurs as a styrene analog in resistant pear genotypes. The His-tagged and affinity chromatography purified E. coli-expressed pear PyPAD1 protein could decarboxylate p-coumaric acid and ferulic acid to p-hydroxystyrene and 3-methoxy-4-hydroxystyrene. In addition, PyPAD1 had the highest activity toward p-coumaric acid. Expression analysis of the PyPAD gene revealed that its expressed as expected, i.e., high when styrene levels and psylla resistance were high.

Keywords: pear Psylla, volatile, GC-MS, resistance

Procedia PDF Downloads 121

Authors: Masuma Novak, Daniel Novak

Abstract:

Background: Sweden along with other Nordic countries has the highest incidence of type 1 diabetes mellitus (T1DM) worldwide. The trend is increasing globally. The diagnosis is often given at the emergency clinic when children arrive with cardinal symptom of T1DM. Children with T1DM are known to have an increased risk of microvascular- and macrovascular complications. A family history of cardiovascular complications may further increase their risk. Clinically evident diabetes-related vascular complications are however rarely visible in childhood and adolescence, whereby an intensive diabetes treatment and normoglycemic control is a goal for every child. This study is a risk evaluation of children with T1DM based on their family’s cardiovascular history. Method: Since 2005 the Better Diabetes Diagnosis (BDD) study is a nationwide Swedish prospective cohort study that recruits new-onset T1DM who are less than 18 years old at time of diagnosis. For each newly diagnosed child, blood samples are collected for specific HLA genotyping and islet autoantibody assays and their family’s cardiovascular history is evaluated. As part of the BDD study, during the years 2010-2013 all children diagnosed with T1DM at the Queen Silvia’s Children’s Hospital in Sweden were asked about their family’s cardiovascular history. Questions regarded maternal and paternal high blood pressure, stroke, and myocardial infarction before the age of 55 years, and hyperlipidemia were answered. A maximum risk score of eight was possible. All children are clinically observed prospectively for early functional and structural abnormalities such as protein uremia, blood pressure, and retinopathy. Results: A total of 275 children aged 0 to 18 years were diagnosed with T1DM at the Queen Silvia’s Children’s Hospital emergency clinic during this four year period. The participation rate was 99.7%. 26.4% of the children had no hereditary cardiovascular risk factors. 22.7 % had one risk factor and 18.8% had two risk factors. 14.8% had three risk factors. 9.7% had four risk factors and 7.5% had five risk factors or more. Conclusion: Among children with T1DM in Sweden there is a difference in hereditary cardiovascular risk factors. These results indicate that children with T1DM who also have increased hereditary cardiovascular risk factors should be monitored closely with early screening for functional and structural cardiovascular abnormalities. This is a very preliminary and ongoing study which will be complemented with the cardiovascular risk analysis among children without T1DM.

Keywords: children, type I diabetes, emergency clinic, CVD risk

Procedia PDF Downloads 340

Authors: S. Pathak, R. Lazarus

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A 25-year-old male HIV patient (CD4 cells 20/µL and HIV viral load 14200000 copies/ml) with a past medical history of duodenal ulcer, pneumocystis carinii pneumonia, oesophageal candidiasis presented with fever and a seizure to hospital. The only recent travel had been a religious pilgrimage from Singapore to Malaysia 5 days prior; during the trip he sustained skin abrasions. The patient had recently started highly active antiretroviral therapy 2 months prior. Clinical examination was unremarkable other than a temperature of 38.8°C and perianal warts. Laboratory tests showed a leukocyte count 12.5x109 cells/L, haemoglobin 9.4 g/dL, normal biochemistry and a C-reactive protein 121 mg/L. CT head and MRI head were unremarkable and cerebrospinal fluid analysis performed after a delay (due to technical difficulties) of 11 days was unremarkable. Blood cultures (three sets) taken on admission showed Gram-negative rods in the anaerobic bottles only at the end of incubation with culture result confirmed by molecular sequencing showing Helicobacter cinaedi. The patient was treated empirically with ceftriaxone for seven days and this was converted to oral co-amoxiclav for a further seven days after the blood cultures became positive. A Transthoracic echocardiogram was unremarkable. The patient made a full recovery. Helicobacter cinaedi is a gram-negative anaerobic fastidious organism affecting patients with comorbidity. Infection may manifest as cellulitius, colitis or as in this case as bloodstream infection – the latter is often attributed to faeco-oral infection. Laboratory identification requires prolonged culture. Therapeutic options may be limited by resistance to macrolides and fluoroquinolones. The likely pathogen inoculation routes in the case described include gastrointestinal translocation due to proctitis at the site of perianal warts, or breach of the skin via abrasions occurring during the pilgrimage. Such organisms are increasing in prevalence as our patient population ages and patients have multiple comorbidities including HIV. It may be necessary in patients with unexplained fever to prolong incubation of sterile sites including blood in order to identify this unusual fastidious organism.

Keywords: fastidious, Helicobacter cinaedi, HIV, immunocompromised

Procedia PDF Downloads 358

Authors: Joyce K. Anastasi, Bernadette Capili

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Community Engagement Strategies to Assist with the Development of an RCT Among People Living with HIV Our research team focuses on developing and testing protocols to manage chronic symptoms. For many years, our team designed and implemented symptom management studies for people living with HIV (PLWH). We identify symptoms that are not curative and are not adequately controlled by conventional therapies. As an exemplar, we describe how we successfully engaged PLWH in developing and refining our research feasibility protocol for distal sensory peripheral neuropathy (DSP) associated with HIV. With input from PLWH with DSP, our research received National Institutes of Health (NIH) research funding support. Significance: DSP is one of the most common neurologic complications in HIV. It is estimated that DSP affects 21% to 50% of PLWH. The pathogenesis of DSP in HIV is complex and unclear. Proposed mechanisms include cytokine dysregulation, viral protein-produced neurotoxicity, and mitochondrial dysfunction associated with antiretroviral medications. There are no FDA-approved treatments for DSP in HIV. Purpose: Aims: 1) to explore the impact of DSP on the lives of PLWH, 2) to identify patients’ perspectives on successful treatments for DSP, 3) to identify interventions considered feasible and sensitive to the needs of PLWH with DSP, and 4) to obtain participant input for protocol/study design. Description of Process: We conducted a needs assessment with PLWH with DSP. From our needs assessment, we learned from the patients’ perspective detailed descriptions of their symptoms; physical functioning with DSP; self-care remedies tried, and desired interventions. We also asked about protocol scheduling, instrument clarity, study compensation, study-related burdens, and willingness to participate in a randomized controlled trial (RCT) with a placebo and a waitlist group. Implications: We incorporated many of the suggestions learned from the need assessment. We developed and completed a feasibility study that provided us with invaluable information that informed subsequent NIH-funded studies. In addition to our extensive clinical and research experience working with PLWH, learning from the patient perspective helped in developing our protocol and promoting a successful plan for recruitment and retention of study participants.

Keywords: clinical trial development, peripheral neuropathy, traditional medicine, HIV, AIDS

Procedia PDF Downloads 60

Authors: Anna Cameron, Chunxia Zhao, Haofei Wang, Yun Liu, Guang Ze Yang

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Since the first publication in 1775, cancer research has built a comprehensive understanding of how cellular components of the tumor niche promote disease development. However, only within the last decade has research begun to establish the impact of non-cellular components of the niche, particularly the extracellular matrix (ECM). The ECM, a three-dimensional scaffold that sustains the tumor microenvironment, plays a crucial role in disease progression. Cancer cells actively deregulate and remodel the ECM to establish a tumor-promoting environment. Recent work has highlighted the need to further our understanding of the complexity of this cancer-ECM relationship. In vitro models use hydrogels to mimic the ECM, as hydrogel matrices offer biological compatibility and stability needed for long term cell culture. However, natural hydrogels are being used in these models verbatim, without tuning their biophysical characteristics to achieve pathophysiological relevance, thus limiting their broad use within cancer research. The biophysical attributes of these gels dictate cancer cell proliferation, invasion, metastasis, and therapeutic response. Evaluating the three most widely used natural hydrogels, Matrigel, collagen, and agarose gel, the permeability, stiffness, and pore-size of each gel were measured and compared to the in vivo environment. The pore size of all three gels fell between 0.5-6 µm, which coincides with the 0.1-5 µm in vivo pore size found in the literature. However, the stiffness for hydrogels able to support cell culture ranged between 0.05 and 0.3 kPa, which falls outside the range of 0.3-20,000 kPa reported in the literature for an in vivo ECM. Permeability was ~100x greater than in vivo measurements, due in large part to the lack of cellular components which impede permeation. Though, these measurements prove important when assessing therapeutic particle delivery, as the ECM permeability decreased with increasing particle size, with 100 nm particles exhibiting a fifth of the permeability of 10 nm particles. This work explores ways of adjusting the biophysical characteristics of hydrogels by changing protein concentration and the trade-off, which occurs due to the interdependence of these factors. The global aim of this work is to produce a more pathophysiologically relevant model for each tumor type.

Keywords: cancer, extracellular matrix, hydrogel, microfluidic

Procedia PDF Downloads 69

Authors: Nahed Mohamed Ismail, Bayomy Mostafa, Ahmed Abdel Kader, Ahmed Mohamed Azzam

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Effect of lining watercourses on the distribution and abundance of fresh water snails at two Egyptian governorates, Baheria (new reclaimed area) and Giza was studied. Seasonal survey in lined and unlined sites during two successive years was carried out. Samples of snails and water were collected from each examined site and the ecological conditions were recorded. The collected snails from each site were placed in plastic aquaria and transferred to the laboratory, where they were sorted out, identified, counted and examined for natural infection. The size frequency distribution was calculated for each snail species. Results revealed that snails were represented in all examined watercourses (lined and unlined) at the two tested habitats by 14 species. (Biomphalaria alexandrina, B. glabrata, Bulinus truncatus, Physa acuta. Helisoma duryi, Lymnaea natalensis, Planorbis planorbis, Cleopatra bulimoids, Lanistes carinatus, Bellamya unicolor, Melanoides tuberculata, Theodoxus nilotica, Succinia cleopatra and Gabbiella senaarensis). During spring, the percentage of live (45%) and dead (55%) snail species was extremely highly significant lower (p>0.001) in lined water bodies compared to the unlined ones (93.5% and 6.5%, respectively) in the examined sites at Baheria. At Giza, the percentage values of live snail species from all lined watercourses (82.6% and 60.2%, during winter and spring, respectively) was significantly lower (p>0.05 & p>0.01) than those in unlined ones (91.1% and 79%, respectively). Size frequency distribution of snails collected from the lined and unlined water bodies at Baheria and Giza governorates during all seasons revealed that during survey, snail populations were stable and the recruitment of young to adult was continuing for some species, where the recruits were observed with adults. However, there was no sign of small snails occurrence in case of B. glabrata and B. alexandrina during autumn, winter and spring and disappear during summer at Giza. Meanwhile they completely absent during all seasons at Baheria Governorate. Chemical analysis of some heavy metals of water samples collected from lined and unlined sites from Baheria and Giza governorates during autumn, winter and spring were approximately as the same in both lined and unlined water bodies. However, Zn and Fe were higher in lined sites (0.78±0.37and 17.4 ± 4.3, respectively) than that of unlined ones (0.4±0.1 and 10.95 ± 1.93, respectively) and Cu was absent in both lined and unlined sites during summer at Baheria governorate. At Giza, Cu and Pb were absent and Fe were higher in lined sites (4.7± 4.2) than that of unlined ones (2.5 ± 1.4) during summer. Statistical analysis showed that no significant difference in all physico-chemical parameters of water in lined and unlined water bodies at the two tested habitats during all seasons. However, it was found that the water conductivity and TDS showed a lower mean values in lined sites than those of unlined ones. Thus, the present obtained data support the concept of utilizing environmental modification such as lining of water courses to help in minimizing the population density of certain vector snails and consequently reduce the transmission of snails born diseases.

Keywords: lining, fresh water, snails, watercourses

Procedia PDF Downloads 230

Authors: Mohamed A. Baba, Gazy Rodowan, Brigita Abakevičienė, Sigitas Tamulevičius, Bartlomiej Lemieszek, Sebastian Molin, Tomas Tamulevičius

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Solid oxide fuel cells (SOFC) efficiently convert hydrogen to energy without producing any disturbances or contaminants. The core of the cell is electrolyte. For improving the performance of electrolyte-supported cells, it is desirable to extend the available exchange surface area by micro-structuring of the electrolyte with laser-based micromachining. This study investigated the electrochemical performance of cells micro machined using a femtosecond laser. Commercial ceramic SOFC (Elcogen, AS) with a total thickness of 400 μm was structured by 1030 nm wavelength Yb: KGW fs-laser Pharos (Light Conversion) using 100 kHz repetition frequency and 290 fs pulse length light by scanning with the galvanometer scanner (ScanLab) and focused with a f-Theta telecentric lens (SillOptics). The sample height was positioned using a motorized z-stage. The microstructures were formed using a laser spiral trepanning in Ni/YSZ anode supported membrane at the central part of the ceramic piece of 5.5 mm diameter at active area of the cell. All surface was drilled with 275 µm diameter holes spaced by 275 µm. The machining processes were carried out under ambient conditions. The microstructural effects of the femtosecond laser treatment on the electrolyte surface were investigated prior to the electrochemical characterisation using a scanning electron microscope (SEM) Quanta 200 FEG (FEI). The Novo control Alpha-A was used for electrochemical impedance spectroscopy on a symmetrical cell configuration with an excitation amplitude of 25 mV and a frequency range of 1 MHz to 0.1 Hz. The fuel cell characterization of the cell was examined on open flanges test setup by Fiaxell. Using nickel mesh on the anode side and au mesh on the cathode side, the cell was electrically linked. The cell was placed in a Kittec furnace with a Process IDentifier temperature controller. The wires were connected to a Solartron 1260/1287 frequency analyzer for the impedance and current-voltage characterization. In order to determine the impact of the anode's microstructure on the performance of the commercial cells, the acquired results were compared to cells with unstructured anode. Geometrical studies verified that the depth of the -holes increased linearly according to laser energy and scanning times. On the other hand, it reduced as the scanning speed increased. The electrochemical analysis demonstrates that the open circuit voltage OCV values of the two cells are equal. Further, the modified cell's initial slope reduces to 0.209 from 0.253 of the unmodified cell, revealing that the surface modification considerably decreases energy loss. Plus, the maximum power density for the cell with the microstructure and the reference cell respectively, are 1.45 and 1.16 Wcm⁻².

Keywords: electrochemical performance, electrolyte-supported cells, laser micro-structuring, solid oxide fuel cells

Procedia PDF Downloads 44

Authors: Lukas J. Evans, Danilo D. Fernando

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Shrub willows (Salix spp.) have many characteristics which make them suitable for a variety of applications such as riparian zone buffers, environmental contaminant sequestration, living snow fences, and biofuel production. In some cases, these functions are limited due to physical or financial obstacles associated with the number of individuals needed to reasonably satisfy that purpose. One way to increase the efficiency of willows is to bioengineer them with the genetic improvements suitable for the desired use. To accomplish this goal, an optimized in vitro transformation protocol via Agrobacterium tumefaciens is necessary to reliably express genes of interest. Therefore, the aim of this study is to observe the influence of tissue culture with different willow cultivars, hormones, and explants on the percentage of calli expressing reporter gene green florescent protein (GFP) to find ideal transformation conditions. Each callus was produced from 1 month old open-pollinated seedlings of three Salix miyabeana cultivars (‘SX61’, ‘WT1’, and ‘WT2’) from three different explants (lamina, petiole, and internodes). Explants were cultured for 1 month on an MS media with different concentrations of 6-Benzylaminopurine (BAP) and 1-Naphthaleneacetic acid (NAA) (No hormones, 1 mg⁻¹L BAP only, 3 mg⁻¹L NAA only, 1 mg⁻¹L BAP and 3 mg⁻¹L NAA, and 3 mg⁻¹L BAP and 1 mg⁻¹L NAA) to produce a callus. Samples were then treated with Agrobacterium tumefaciens at an OD600 of 0.6-0.8 to insert the transgene GFP for 30 minutes, co-cultivated for 72 hours, and selected on the same media type they were cultured on with added 7.5 mg⁻¹L of Hygromycin for 1 week before GFP visualization under a UV dissecting scope. Percentage of GFP expressing calli as well as the average number of fluorescing GFP units per callus were recorded and results were evaluated through an ANOVA test (α = 0.05). The WT1 internode-derived calli on media with 3 mg-1L NAA+1 mg⁻¹L BAP and mg⁻¹L BAP alone produced a significantly higher percentage of GFP expressing calli than each other group (19.1% and 19.4%, respectively). Additionally, The WT1 internode group cultured with 3 mg⁻¹L NAA+1 mg⁻¹L BAP produced an average of 2.89 GFP units per callus while the group cultivated with 1 mg⁻¹L BAP produced an average of 0.84 GFP units per callus. In conclusion, genotype, explant choice, and hormones all play a significant role in increasing successful transformation in willows. Future studies to produce whole callus GFP expression and subsequent plantlet regeneration are necessary for a complete willow transformation protocol.

Keywords: agrobacterium, callus, Salix, tissue culture

Procedia PDF Downloads 101

Authors: Adnan A. Kadi, Nasser S. Al-Shakliah, Haitham Al-Rabiah

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Zorifertinib is a novel, potent, oral, a small molecule used to treat non-small cell lung cancer (NSCLC). zorifertinib is an Epidermal Growth Factor Receptor (EGFR) inhibitor and has good blood–brain barrier permeability for (NSCLC) patients with EGFR mutations. zorifertinibis currently at phase II/III clinical trials. The current research reports the characterization and identification of in vitro, in vivo and reactive intermediates of zorifertinib. Prediction of susceptible sites of metabolism and reactivity pathways (cyanide and GSH) of zorifertinib were performed by the Xenosite web predictor tool. In-vitro metabolites of zorifertinib were performed by incubation with rat liver microsomes (RLMs) and isolated perfused rat liver hepatocytes. Extraction of zorifertinib and it's in vitro metabolites from the incubation mixtures were done by protein precipitation. In vivo metabolism was done by giving a single oral dose of zorifertinib(10 mg/Kg) to Sprague Dawely rats in metabolic cages by using oral gavage. Urine was gathered and filtered at specific time intervals (0, 6, 12, 18, 24, 48, 72,96and 120 hr) from zorifertinib dosing. A similar volume of ACN was added to each collected urine sample. Both layers (organic and aqueous) were injected into liquid chromatography ion trap mass spectrometry(LC-IT-MS) to detect vivozorifertinib metabolites. N-methyl piperizine ring and quinazoline group of zorifertinib undergoe metabolism forming iminium and electro deficient conjugated system respectively, which are very reactive toward nucleophilic macromolecules. Incubation of zorifertinib with RLMs in the presence of 1.0 mM KCN and 1.0 Mm glutathione were made to check reactive metabolites as it is often responsible for toxicities associated with this drug. For in vitro metabolites there were nine in vitro phase I metabolites, four in vitro phase II metabolites, eleven reactive metabolites(three cyano adducts, five GSH conjugates metabolites, and three methoxy metabolites of zorifertinib were detected by LC-IT-MS. For in vivo metabolites, there were eight in vivo phase I, tenin vivo phase II metabolitesofzorifertinib were detected by LC-IT-MS. In vitro and in vivo phase I metabolic pathways wereN- demthylation, O-demethylation, hydroxylation, reduction, defluorination, and dechlorination. In vivo phase II metabolic reaction was direct conjugation of zorifertinib with glucuronic acid and sulphate.

Keywords: in vivo metabolites, in vitro metabolites, cyano adducts, GSH conjugate

Procedia PDF Downloads 173

Authors: Stamatia Nasiakou, Dimitrios Chouvardas, Michael Vrahnakis, Vassiliki Kleftoyanni

Abstract:

Recent research in the mountainous and semi-mountainous rural landscapes of Greece shows that they have been significantly changed over the last 80 years. These changes have the form of structural modification of land cover/use patterns, with the main characteristic being the extensive expansion of dense forests and shrubs at the expense of grasslands and extensive agricultural areas. The aim of this research was to study the 60-year changes (1960-2019) of land cover/ use units in the rural landscape of Mouzaki (Karditsa Prefecture, central Greece). Relevant cartographic material such as forest land use maps, digital maps (Corine Land Cover -2018), 1960 aerial photos from Hellenic Military Geographical Service, and satellite imagery (Google Earth Pro 2014, 2016, 2017 and 2019) was collected and processed in order to study landscape evolution. ArcGIS v 10.2.2 software was used to process the cartographic material and to produce several sets of data. Main product of the analysis was a digitized photo-mosaic of the 1960 aerial photographs, a digitized photo-mosaic of recent satellite images (2014, 2016, 2017 and 2019), and diagrams and maps of temporal transformation of the rural landscape (1960 – 2019). Maps and diagrams were produced by applying photointerpretation techniques and a suitable land cover/ use classification system on the two photo-mosaics. Demographic and socioeconomic inventory data was also collected mainly from diachronic census reports of the Hellenic Statistical Authority and local sources. Data analysis of the temporal transformation of land cover/ use units showed that they are mainly located in the central and south-eastern part of the study area, which mainly includes the mountainous part of the landscape. The most significant change is the expansion of the dense forests that currently dominate the southern and eastern part of the landscape. In conclusion, the produced diagrams and maps of the land cover/ use evolution suggest that woody vegetation in the rural landscape of Mouzaki has significantly increased over the past 60 years at the expense of the open areas, especially grasslands and agricultural areas. Demographic changes, land abandonment and the transformation of traditional farming practices (e.g. agroforestry) were recognized as the main cause of the landscape change. This study is part of a broader research project entitled “Perspective of Agroforestry in Thessaly region: A research on social, environmental and economic aspects to enhance farmer participation”. The project is funded by the General Secretariat for Research and Technology (GSRT) and the Hellenic Foundation for Research and Innovation (HFRI).

Keywords: Agroforestry, Forest expansion, Land cover/ use changes, Mountainous and semi-mountainous areas

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Authors: S. Fröhlich, M. Herold, M. Allmer

Abstract:

Early stage quantitative analysis of host cell protein (HCP) variations is challenging yet necessary for comprehensive bioprocess development. High resolution mass spectrometry (HRMS) provides a high-end technology for accurate identification alongside with quantitative information. Hereby we describe a flexible HRMS assay platform to quantify HCPs relevant in microbial expression systems such as E. Coli in both up and downstream development by means of MVDA tools. Cell pellets were lysed and proteins extracted, purified samples not further treated before applying the SMART tryptic digest kit. Peptides separation was optimized using an RP-UHPLC separation platform. HRMS-MSMS analysis was conducted on an Orbitrap Velos Elite applying CID. Quantification was performed label-free taking into account ionization properties and physicochemical peptide similarities. Results were analyzed using SIEVE 2.0 (Thermo Fisher Scientific) and SIMCA (Umetrics AG). The developed HRMS platform was applied to an E. Coli expression set with varying productivity and the corresponding downstream process. Selected HCPs were successfully quantified within the fmol range. Analysing HCP networks based on pattern analysis facilitated low level quantification and enhanced validity. This approach is of high relevance for high-throughput screening experiments during upstream development, e.g. for titer determination, dynamic HCP network analysis or product characterization. Considering the downstream purification process, physicochemical clustering of identified HCPs is of relevance to adjust buffer conditions accordingly. However, the technology provides an innovative approach for label-free MS based quantification relying on statistical pattern analysis and comparison. Absolute quantification based on physicochemical properties and peptide similarity score provides a technological approach without the need of sophisticated sample preparation strategies and is therefore proven to be straightforward, sensitive and highly reproducible in terms of product characterization.

Keywords: process analytical technology, mass spectrometry, process characterization, MVDA, pattern recognition

Procedia PDF Downloads 231

Authors: Obuzor Eze Obuzor, Ekpoke Okurube Sliver

Abstract:

The study was carried out at Debee farms at Ahoada West Local Government area, Rivers State, Nigeria. To evaluate the impact of two different cost-effective available feed composition on growth performance of weaned piglets. Thirty weaned uncontrolled cross bred (Large white x pietrain) piglets of average initial weight of 3.04 Kg weaned at 30days were assigned to three dietary treatments, comprising three replicates of 10 weaned piglets each, piglets were kept at 7 °C in different pens with dimensions of 4.50 × 4.50 m. The design of the experiment was completely randomized design, data from the study were subjected to one-way analysis of variance (ANOVA) and significant means were separated using Duncan's Multiple Range Test using Statistical Analysis System (SAS) software for windows (2 0 0 3), statistical significance was assessed at P < 0.05 (95% confidence interval) while survival rate was calculated using simple percentage. A standard diet was prepared to meet the nutrient requirements of weaned piglets at (20.8% crude protein). The three diets were fed to the animals in concrete feeding trough, control diet (C) had soybean meal while first treatment had spent grain (T1) and the second treatment had wheat offal (T2) respectively. The experiment was partitioned into four weeks periods (days 1-7, 8-14, 15-21 and 22-28). Feed and water were given unrestrictedly throughout the period of the experiment. The feed intake and weights of the pigs were recorded on weekly basis. Feed conversion ratio and daily weight gain were calculated and the study lasted for four weeks. There was no significant (P>0.05) effect of diet on survival rate, final body weight, average daily weight gain, daily feed intake and feed conversion ratio. The overall performance showed that treatment one (T1) had survival rate (93%), improved daily weight gain (36.21 g), average daily feed intake (120.14 g) and had the best feed conversion ratio (0.29) similar high mean value with the control while treatment two (T2) had lowest and negative response to all parameters. It could be concluded that feed formulated with spent grain is cheaper than control (soybean meal) and also improved the growth performance of weaned piglets.

Keywords: piglets, weaning, feed conversions ratio, daily weight gain

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Authors: Nilgün Öztürk, Hakan Sabahtin Ali, Hülya Tuba Kıyan

Abstract:

The genus Chenopodium belongs to Amaranthaceae, is represented by approximately 250 species in the world and 15 species and three subspecies in Turkey. Chenopodium species are traditionally used to treat chest and abdominal pain, shortness of breath, cough and neurological disorders. Chenopodium quinoa Willd. (Quinoa) is native to Andes region of South America (especially Peru and Bolivia) and cultivated in many countries include also Turkey in the world nowadays. The seeds of quinoa are rich in protein, and the phytochemical composition consists of antioxidant substances such as polyphenolic compounds, flavonoids, vitamins, and minerals; anticancer and neuroprotective compounds such as tocotrienols; anti-inflammatory compounds such as carotenoids and anthocyanins and also saponins and starch. Food products of quinoa such as quinoa cereal bar, pasta and cornflakes are used in the diet made during many disorders like obesity, cardiovascular disorder, hypertension and Celiac disease. Also quinoa seems to have antimicrobial, anti-inflammatory and cholesterol-lowering properties because of its bioactive compounds. In this present study, the aqueous ethanolic extracts of the seeds of three different coloured genotypes of quinoa were investigated for their antioxidant activities using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, ferrous ion-chelating effect, ferric-reducing antioxidant power, ABTS radical cation decolorization assays and total phenolic contents using Folin-Ciocalteu assay. Among the three genotypes of quinoa; the aqueous ethanolic extract of the red genotype had the highest total phenolic content (83.54 ± 2.12 mg gallic acid/100 g extract) whereas the extract of the white genotype had the lowest total phenolic content (70.66 ± 0.25 mg gallic acid/100 g). According to the antioxidant activity results; the extracts showed moderate reducing power effect whereas weak ABTS radical cation decolorization and ferrous ion-chelating effect and also too weak DPPH radical scavenging activity when compared to the positive standards.

Keywords: amaranthaceae, antioxidant activity, Chenopodium quinoa willd., total phenolic content

Procedia PDF Downloads 159