Search results for: protein synthesis
Commenced in January 2007
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Edition: International
Paper Count: 4359

Search results for: protein synthesis

549 Recursion, Merge and Event Sequence: A Bio-Mathematical Perspective

Authors: Noury Bakrim

Abstract:

Formalization is indeed a foundational Mathematical Linguistics as demonstrated by the pioneering works. While dialoguing with this frame, we nonetheless propone, in our approach of language as a real object, a mathematical linguistics/biosemiotics defined as a dialectical synthesis between induction and computational deduction. Therefore, relying on the parametric interaction of cycles, rules, and features giving way to a sub-hypothetic biological point of view, we first hypothesize a factorial equation as an explanatory principle within Category Mathematics of the Ergobrain: our computation proposal of Universal Grammar rules per cycle or a scalar determination (multiplying right/left columns of the determinant matrix and right/left columns of the logarithmic matrix) of the transformable matrix for rule addition/deletion and cycles within representational mapping/cycle heredity basing on the factorial example, being the logarithmic exponent or power of rule deletion/addition. It enables us to propone an extension of minimalist merge/label notions to a Language Merge (as a computing principle) within cycle recursion relying on combinatorial mapping of rules hierarchies on external Entax of the Event Sequence. Therefore, to define combinatorial maps as language merge of features and combinatorial hierarchical restrictions (governing, commanding, and other rules), we secondly hypothesize from our results feature/hierarchy exponentiation on graph representation deriving from Gromov's Symbolic Dynamics where combinatorial vertices from Fe are set to combinatorial vertices of Hie and edges from Fe to Hie such as for all combinatorial group, there are restriction maps representing different derivational levels that are subgraphs: the intersection on I defines pullbacks and deletion rules (under restriction maps) then under disjunction edges H such that for the combinatorial map P belonging to Hie exponentiation by intersection there are pullbacks and projections that are equal to restriction maps RM₁ and RM₂. The model will draw on experimental biomathematics as well as structural frames with focus on Amazigh and English (cases from phonology/micro-semantics, Syntax) shift from Structure to event (especially Amazigh formant principle resolving its morphological heterogeneity).

Keywords: rule/cycle addition/deletion, bio-mathematical methodology, general merge calculation, feature exponentiation, combinatorial maps, event sequence

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548 Let-7 Mirnas Regulate Inflammatory Cytokine Production in Bovine Endometrial Cells after Lipopolysaccharide Challenge by Targeting TNFα

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

Abstract:

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

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547 Irradiated-Chitosan and Methyl Jasmonate Modulate the Growth, Physiology and Alkaloids Production in Catharanthus roseus (l.) G. Don.

Authors: Moin Uddin, M. Masroor A. Khan, Faisal Rasheed, Tariq Ahmad Dar, Akbar Ali, Lalit Varshney

Abstract:

Oligomers, obtained by exposing the natural polysaccharides (alginate, carrageenan, chitosan, etc.) to cobalt-60 generated gamma radiation may prove as potent plant growth promoters when applied as foliar sprays to the plants. They function as endogenous growth elicitors, triggering the synthesis of different enzymes and modulating various plant responses by exploiting the gene expression. Exogenous application of Jasmonic acid or of its methyl ester, methyl jasmonate (MeJ) has been reported to increase the secondary metabolites production in medicinal and aromatic plants. Keeping this in mind, three pot experiments were conducted to test whether the foliar application of irradiated-chitosan (IC) and MeJ, applied alone or in combination, could augment the active constituents as well as growth, physiological and yield attributes of Catharanthus roseus, which carries anticancer alkaloids, viz. vincristine and vinblastine, in its leaves in addition to various other useful alkaloids. Totally, 5 spray treatments, comprising various aqueous solutions of IC [20, 40, 80 and 160 mg L-1 (Experiment 1)], MeJ (10, 20, 30 and 40 mg L-1 (Experiment 2)] and those of IC+MeJ [40+20, 40+30, 80+20, 80+30, 160+20 and 160+30 mg L-1 (Experiment 3)], were applied at seven days interval. Total leaf-alkaloids content as well as growth, physiological and yield parameters, evaluated at 120 days after sowing, were significantly enhanced by IC application. IC application could not increase the leaf-content of vincristine and vinblastine; nonetheless, it significantly augmented the yield of these alkaloids owing to enhancing the dry mass of leaves per plant. MeJ application, particularly at 30 mg L-1, increased both content (17%) and yield (48%) of total leaf-alkaloids as well as the content and yield of vincristine ( 29 and 63%, respectively) and vinblastine (14 and 44%, respectively) alkaloids, though it significantly decreased most other parameters studied, particularly at higher concentrations (30 and 40 mg L-1 of MeJ). As compared to the control (water-spray treatment), collective application of IC (80 mg L-1) and MeJ (20 mg L-1) resulted in the highest values of most of the parameters studied. However, 80 mg L-1 of IC applied with 30 mg L-1 of MeJ gave the best results for the content and yield of total as well as anticancer leaf-alkaloids (vincristine and vinblastine). Comparing the control, it increased the content and yield of total leaf-alkaloids (37 and 118%, respectively) and those of vincristine (65 and 163%, respectively) and vinblastine (31 and 107%, respectively). Conclusively, the applied technique significantly enhanced the production of total as well as anticancer alkaloids of Catharanthus roseus.

Keywords: anticancer alkaloids (vincristine and vinblastine), catharanthus roseus, irradiated chitosan, methyl jasmonate

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546 Intracellular Sphingosine-1-Phosphate Receptor 3 Contributes to Lung Tumor Cell Proliferation

Authors: Michela Terlizzi, Chiara Colarusso, Aldo Pinto, Rosalinda Sorrentino

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Sphingosine-1-phosphate (S1P) is a membrane-derived bioactive phospholipid exerting a multitude of effects on respiratory cell physiology and pathology through five S1P receptors (S1PR1-5). Higher levels of S1P have been registered in a broad range of respiratory diseases, including inflammatory disorders and cancer, although its exact role is still elusive. Based on our previous study in which we found that S1P/S1PR3 is involved in an inflammatory pattern via the activation of Toll-like Receptor 9 (TLR9), highly expressed on lung cancer cells, the main goal of the current study was to better understand the involvement of S1P/S1PR3 pathway/signaling during lung carcinogenesis, taking advantage of a mouse model of first-hand smoke exposure and of carcinogen-induced lung cancer. We used human samples of Non-Small Cell Lung Cancer (NSCLC), a mouse model of first-hand smoking, and of Benzo(a)pyrene (BaP)-induced tumor-bearing mice and A549 lung adenocarcinoma cells. We found that the intranuclear, but not the membrane, localization of S1PR3 was associated to the proliferation of lung adenocarcinoma cells, the mechanism that was correlated to human and mouse samples of smoke-exposure and carcinogen-induced lung cancer, which were characterized by higher utilization of S1P. Indeed, the inhibition of the membrane S1PR3 did not alter tumor cell proliferation after TLR9 activation. Instead, according to the nuclear localization of sphingosine kinase (SPHK) II, the enzyme responsible for the catalysis of the S1P last step synthesis, the inhibition of the kinase completely blocked the endogenous S1P-induced tumor cell proliferation. These results prove that the endogenous TLR9-induced S1P can on one side favor pro-inflammatory mechanisms in the tumor microenvironment via the activation of cell surface receptors, but on the other tumor progression via the nuclear S1PR3/SPHK II axis, highlighting a novel molecular mechanism that identifies S1P as one of the crucial mediators for lung carcinogenesis-associated inflammatory processes and that could provide differential therapeutic approaches especially in non-responsive lung cancer patients.

Keywords: sphingosine-1-phosphate (S1P), S1P Receptor 3 (S1PR3), smoking-mice, lung inflammation, lung cancer

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545 Assessment of the Physical Activity Level and the Nutritional Status among Students in Bowen University, Iwo, Osun State, Nigeria

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

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544 The Effects of Drought and Nitrogen on Soybean (Glycine max (L.) Merrill) Physiology and Yield

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

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543 Estimation of Carbon Losses in Rice: Wheat Cropping System of Punjab, Pakistan

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

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542 Core-Shell Nanofibers for Prevention of Postsurgical Adhesion

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

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541 Dry-Extrusion of Asian Carp, a Sustainable Source of Natural Methionine for Organic Poultry Production

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

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540 Assessment of Cattle Welfare Traveling Long Distance from Jessore (Indian Border) to Chittagong, Bangladesh

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

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539 Determination of the Vaccine Induced Immunodominant Regions of Nucleoprotein Crimean-Congo Hemorrhagic Fever Virus

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

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538 Profiling the Volatile Metabolome in Pear Leaves with Different Resistance to the Pear Psylla Cacopsylla bidens (Sulc) and Characterization of Phenolic Acid Decarboxylase

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

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537 Existing Cardiovascular Risk among Children Diagnosed with Type 1 Diabetes Mellitus at the Emergency Clinic

Authors: Masuma Novak, Daniel Novak

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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

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536 Fastidious Enteric Pathogens in HIV

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 373
535 Community Engagement Strategies to Assist with the Development of an RCT Among People Living with HIV

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 81
534 Synthesis of Iron Oxide Nanoparticles Using Different Stabilizers and Study of Their Size and Properties

Authors: Mohammad Hassan Ramezan zadeh 1 , Majid Seifi 2 , Hoda Hekmat ara 2 1Biomedical Engineering Department, Near East University, Nicosia, Cyprus 2Physics Department, Guilan University , P.O. Box 41335-1914, Rasht, Iran.

Abstract:

Magnetic nano particles of ferric chloride were synthesised using a co-precipitation technique. For the optimal results, ferric chloride at room temperature was added to different surfactant with different ratio of metal ions/surfactant. The samples were characterised using transmission electron microscopy, X-ray diffraction and Fourier transform infrared spectrum to show the presence of nanoparticles, structure and morphology. Magnetic measurements were also carried out on samples using a Vibrating Sample Magnetometer. To show the effect of surfactant on size distribution and crystalline structure of produced nanoparticles, surfactants with various charge such as anionic cetyl trimethyl ammonium bromide (CTAB), cationic sodium dodecyl sulphate (SDS) and neutral TritonX-100 was employed. By changing the surfactant and ratio of metal ions/surfactant the size and crystalline structure of these nanoparticles were controlled. We also show that using anionic stabilizer leads to smallest size and narrowest size distribution and the most crystalline (polycrystalline) structure. In developing our production technique, many parameters were varied. Efforts at reproducing good yields indicated which of the experimental parameters were the most critical and how carefully they had to be controlled. The conditions reported here were the best that we encountered but the range of possible parameter choice is so large that these probably only represent a local optimum. The samples for our chemical process were prepared by adding 0.675 gr ferric chloride (FeCl3, 6H2O) to three different surfactant in water solution. The solution was sonicated for about 30 min until a transparent solution was achieved. Then 0.5 gr sodium hydroxide (NaOH) as a reduction agent was poured to the reaction drop by drop which resulted to participate reddish brown Fe2O3 nanoparticles. After washing with ethanol the obtained powder was calcinated in 600°C for 2h. Here, the sample 1 contained CTAB as a surfactant with ratio of metal ions/surfactant 1/2, sample 2 with CTAB and ratio 1/1, sample 3 with SDS and ratio 1/2, sample 4 SDS 1/1, sample 5 is triton-X-100 with 1/2 and sample 6 triton-X-100 with 1/1.

Keywords: iron oxide nanoparticles, stabilizer, co-precipitation, surfactant

Procedia PDF Downloads 246
533 Engineering a Tumor Extracellular Matrix Towards an in vivo Mimicking 3D Tumor Microenvironment

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 89
532 The Microstructure and Corrosion Behavior of High Entropy Metallic Layers Electrodeposited by Low and High-Temperature Methods

Authors: Zbigniew Szklarz, Aldona Garbacz-Klempka, Magdalena Bisztyga-Szklarz

Abstract:

Typical metallic alloys bases on one major alloying component, where the addition of other elements is intended to improve or modify certain properties, most of all the mechanical properties. However, in 1995 a new concept of metallic alloys was described and defined. High Entropy Alloys (HEA) contains at least five alloying elements in an amount from 5 to 20 at.%. A common feature this type of alloys is an absence of intermetallic phases, high homogeneity of the microstructure and unique chemical composition, what leads to obtaining materials with very high strength indicators, stable structures (also at high temperatures) and excellent corrosion resistance. Hence, HEA can be successfully used as a substitutes for typical metallic alloys in various applications where a sufficiently high properties are desirable. For fabricating HEA, a few ways are applied: 1/ from liquid phase i.e. casting (usually arc melting); 2/ from solid phase i.e. powder metallurgy (sintering methods preceded by mechanical synthesis) and 3/ from gas phase e.g. sputtering or 4/ other deposition methods like electrodeposition from liquids. Application of different production methods creates different microstructures of HEA, which can entail differences in their properties. The last two methods also allows to obtain coatings with HEA structures, hereinafter referred to as High Entropy Films (HEF). With reference to above, the crucial aim of this work was the optimization of the manufacturing process of the multi-component metallic layers (HEF) by the low- and high temperature electrochemical deposition ( ED). The low-temperature deposition process was crried out at ambient or elevated temperature (up to 100 ᵒC) in organic electrolyte. The high-temperature electrodeposition (several hundred Celcius degrees), in turn, allowed to form the HEF layer by electrochemical reduction of metals from molten salts. The basic chemical composition of the coatings was CoCrFeMnNi (known as Cantor’s alloy). However, it was modified by other, selected elements like Al or Cu. The optimization of the parameters that allow to obtain as far as it possible homogeneous and equimolar composition of HEF is the main result of presented studies. In order to analyse and compare the microstructure, SEM/EBSD, TEM and XRD techniques were employed. Morover, the determination of corrosion resistance of the CoCrFeMnNi(Cu or Al) layers in selected electrolytes (i.e. organic and non-organic liquids) was no less important than the above mentioned objectives.

Keywords: high entropy alloys, electrodeposition, corrosion behavior, microstructure

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531 A Comparative Study on the Positive and Negative of Electronic Word-of-Mouth on the SERVQUAL Scale-Take A Certain Armed Forces General Hospital in Taiwan As An Example

Authors: Po-Chun Lee, Li-Lin Liang, Ching-Yuan Huang

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Purpose: Research on electronic word-of-mouth (eWOM)& online review has been widely used in service industry management research in recent years. The SERVQUAL scale is the most commonly used method to measure service quality. Therefore, the purpose of this research is to combine electronic word of mouth & online review with the SERVQUAL scale. To explore the comparative study of positive and negative electronic word-of-mouth reviews of a certain armed force general hospital in Taiwan. Data sources: This research obtained online word-of-mouth comment data on google maps from a military hospital in Taiwan in the past ten years through Internet data mining technology. Research methods: This study uses the semantic content analysis method to classify word-of-mouth reviews according to the revised PZB SERVQUAL scale. Then carry out statistical analysis. Results of data synthesis: The results of this study disclosed that the negative reviews of this military hospital in Taiwan have been increasing year by year. Under the COVID-19 epidemic, positive word-of-mouth has a downward trend. Among the five determiners of SERVQUAL of PZB, positive word-of-mouth reviews performed best in “Assurance,” with a positive review rate of 58.89%, Followed by 43.33% of “Responsiveness.” In negative word-of-mouth reviews, “Assurance” performed the worst, with a positive rate of 70.99%, followed by responsive 29.01%. Conclusions: The important conclusions of this study disclosed that the total number of electronic word-of-mouth reviews of the military hospital has revealed positive growth in recent years, and the positive word-of-mouth growth has revealed negative growth after the epidemic of COVID-19, while the negative word-of-mouth has grown substantially. Regardless of the positive and negative comments, what patients care most about is “Assurance” of the professional attitude and skills of the medical staff, which needs to be strengthened most urgently. In addition, good “Reliability” will help build positive word-of-mouth. However, poor “Responsiveness” can easily lead to the spread of negative word-of-mouth. This study suggests that the hospital should focus on these few service-oriented quality management and audits.

Keywords: quality of medical service, electronic word-of-mouth, armed forces general hospital

Procedia PDF Downloads 174
530 Influence of Genotype, Explant, and Hormone Treatment on Agrobacterium-Transformation Success in Salix Callus Culture

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

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529 Identification and Characterization of in Vivo, in Vitro and Reactive Metabolites of Zorifertinib Using Liquid Chromatography Lon Trap Mass Spectrometry

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 193
528 Novel Adomet Analogs as Tools for Nucleic Acids Labeling

Authors: Milda Nainyte, Viktoras Masevicius

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Biological methylation is a methyl group transfer from S-adenosyl-L-methionine (AdoMet) onto N-, C-, O- or S-nucleophiles in DNA, RNA, proteins or small biomolecules. The reaction is catalyzed by enzymes called AdoMet-dependent methyltransferases (MTases), which represent more than 3 % of the proteins in the cell. As a general mechanism, the methyl group from AdoMet replaces a hydrogen atom of nucleophilic center producing methylated DNA and S-adenosyl-L-homocysteine (AdoHcy). Recently, DNA methyltransferases have been used for the sequence-specific, covalent labeling of biopolymers. Two types of MTase catalyzed labeling of biopolymers are known, referred as two-step and one-step. During two-step labeling, an alkylating fragment is transferred onto DNA in a sequence-specific manner and then the reporter group, such as biotin, is attached for selective visualization using suitable chemistries of coupling. This approach of labeling is quite difficult and the chemical hitching does not always proceed at 100 %, but in the second step the variety of reporter groups can be selected and that gives the flexibility for this labeling method. In the one-step labeling, AdoMet analog is designed with the reporter group already attached to the functional group. Thus, the one-step labeling method would be more comfortable tool for labeling of biopolymers in order to prevent additional chemical reactions and selection of reaction conditions. Also, time costs would be reduced. However, effective AdoMet analog appropriate for one-step labeling of biopolymers and containing cleavable bond, required for reduction of PCR interferation, is still not known. To expand the practical utility of this important enzymatic reaction, cofactors with activated sulfonium-bound side-chains have been produced and can serve as surrogate cofactors for a variety of wild-type and mutant DNA and RNA MTases enabling covalent attachment of these chains to their target sites in DNA, RNA or proteins (the approach named methyltransferase-directed Transfer of Activated Groups, mTAG). Compounds containing hex-2-yn-1-yl moiety has proved to be efficient alkylating agents for labeling of DNA. Herein we describe synthetic procedures for the preparation of N-biotinoyl-N’-(pent-4-ynoyl)cystamine starting from the coupling of cystamine with pentynoic acid and finally attaching the biotin as a reporter group. The synthesis of the first AdoMet based cofactor containing a cleavable reporter group and appropriate for one-step labeling was developed.

Keywords: adoMet analogs, DNA alkylation, cofactor, methyltransferases

Procedia PDF Downloads 193
527 Statistical Pattern Recognition for Biotechnological Process Characterization Based on High Resolution Mass Spectrometry

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

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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 245
526 Effect of Different Feed Composition on the Growth Performance in Early Weaned Piglets

Authors: Obuzor Eze Obuzor, Ekpoke Okurube Sliver

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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

Procedia PDF Downloads 63
525 Determination of Phenolic Contents and Antioxidant Activities of Chenopodium quinoa Willd. Seed Extracts

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

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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 176
524 Design of Identification Based Adaptive Control for Fermentation Process in Bioreactor

Authors: J. Ritonja

Abstract:

The biochemical technology has been developing extremely fast since the middle of the last century. The main reason for such development represents a requirement for large production of high-quality biologically manufactured products such as pharmaceuticals, foods, and beverages. The impact of the biochemical industry on the world economy is enormous. The great importance of this industry also results in intensive development in scientific disciplines relevant to the development of biochemical technology. In addition to developments in the fields of biology and chemistry, which enable to understand complex biochemical processes, development in the field of control theory and applications is also very important. In the paper, the control for the biochemical reactor for the milk fermentation was studied. During the fermentation process, the biophysical quantities must be precisely controlled to obtain the high-quality product. To control these quantities, the bioreactor’s stirring drive and/or heating system can be used. Available commercial biochemical reactors are equipped with open loop or conventional linear closed loop control system. Due to the outstanding parameters variations and the partial nonlinearity of the biochemical process, the results obtained with these control systems are not satisfactory. To improve the fermentation process, the self-tuning adaptive control system was proposed. The use of the self-tuning adaptive control is suggested because the parameters’ variations of the studied biochemical process are very slow in most cases. To determine the linearized mathematical model of the fermentation process, the recursive least square identification method was used. Based on the obtained mathematical model the linear quadratic regulator was tuned. The parameters’ identification and the controller’s synthesis are executed on-line and adapt the controller’s parameters to the fermentation process’ dynamics during the operation. The use of the proposed combination represents the original solution for the control of the milk fermentation process. The purpose of the paper is to contribute to the progress of the control systems for the biochemical reactors. The proposed adaptive control system was tested thoroughly. From the obtained results it is obvious that the proposed adaptive control system assures much better following of the reference signal as a conventional linear control system with fixed control parameters.

Keywords: adaptive control, biochemical reactor, linear quadratic regulator, recursive least square identification

Procedia PDF Downloads 119
523 Evaluation of Nutrient Intake, Body Weight Gain and Carcass Characteristics of Growing Washera Lamb Fed Grass Hay as a Basal Diet with Supplementation of Dried Atella and Niger Seed Cake in Different Combinations

Authors: Fana Woldetsadik

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Ethiopia has a huge livestock population, including sheep, that has been contributing a considerable portion to the economy of the country and still promising to rally around the economic advancement of the country. However, feed shortage is a limiting factor in the production and productivity of sheep among Ethiopian smallholder farmers. Therefore, the aim of this study was to prove the role of the locally available brewery by-products called dried Atella as a supplement in feed intake, digestibility, live weight gain, carcass yield, and economic benefit in comparison with commercially purchased supplements known as niger seed cake (NSC). This on-station feeding experiment was conducted on the Zenzelma Campus of Bahir Dar University animal farm. The experimental design used for this research was a completely randomized design (CRD) with five replications. The crude protein (CP) content of dried Atella, wheat bran (WB), natural pasture hay (NPH) and NSC were about 25.07%, 16.57%, 4.48% and 38.04%, respectively, while the neutral detergent fibre (NDF),acid detergent fibre (ADF) and acid detergent lignin (ADL) content of dried Atella, WB, NPH and NSC were around 31.75%, 8.31%, 8.14%; 42.05%, 22.64%, 4.04%; 74.21%, 50.81%, 8.66%; 42.31%, 26.95% and 6.9%, respectively. The result depicted that a higher(P < 0.001) feed intake, nutrient intake, and digestibility for lambs supplemented with Atella than those supplemented with NSC. Furthermore, daily body weight gain and carcass characteristics were better (P < 0.05) for the sheep supplemented with dried Atella than NSC. On the other hand, in terms of profitability, although there was no substantial difference (P > 0.05) between T2 (animals fed NPH,NSC and WB) and T3 (animals fed NPH, Atella and WB), slightly better benefit was recorded in T3 groups. However, loss of money was recorded in T1 (animals fed NPH and WB). Hence, from the biological performance of lambs, it was concluded that Atella could be a potential supplementary feed for sheep fattening among smallholder farmers than NSC despite no profitability difference. Nevertheless, further investigation is recommended to examine the consequence of supplementation of NPH with NSC and NPH with Atella on fatty acid profile analysis, the physicochemical composition of meat, and meat composition.

Keywords: Attela, Bahir Dar university, Carcass yield, digestibility, natural pasture hay, Niger seed cake, smallholder farmers, weight gain, Ethiopia

Procedia PDF Downloads 147
522 Potential Serological Biomarker for Early Detection of Pregnancy in Cows

Authors: Shveta Bathla, Preeti Rawat, Sudarshan Kumar, Rubina Baithalu, Jogender Singh Rana, Tushar Kumar Mohanty, Ashok Kumar Mohanty

Abstract:

Pregnancy is a complex process which includes series of events such as fertilization, formation of blastocyst, implantation of embryo, placental formation and development of fetus. The success of these events depends on various interactions which are synchronized by endocrine interaction between a receptive dam and competent embryo. These interactions lead to change in expression of hormones and proteins. But till date no protein biomarker is available which can be used to detect successful completion of these events. We employed quantitative proteomics approach to develop putative serological biomarker which has diagnostic applicability for early detection of pregnancy in cows. For this study, sera were collected from control (non-pregnant, n=6) and pregnant animals on successive days of pregnancy (7, 19, 45, n=6). The sera were subjected to depletion for removal of albumin using Norgen depletion kit. The tryptic peptides were labeled with iTRAQ. The peptides were pooled and fractionated using bRPLC over 80 min gradient. Then 12 fractions were injected to nLC for identification and quantitation in DDA mode using ESI. Identification using Mascot search revealed 2056 proteins out of which 352 proteins were differentially expressed. Twenty proteins were upregulated and twelve proteins were down-regulated with fold change > 1.5 and < 0.6 respectively (p < 0.05). The gene ontology studies of DEPs using Panther software revealed that majority of proteins are actively involved in catalytic activities, binding and enzyme regulatory activities. The DEP'S such as NF2, MAPK, GRIPI, UGT1A1, PARP, CD68 were further subjected to pathway analysis using KEGG and Cytoscape plugin Cluego that showed involvement of proteins in successful implantation, maintenance of pluripotency, regulation of luteal function, differentiation of endometrial macrophages, protection from oxidative stress and developmental pathways such as Hippo. Further efforts are continuing for targeted proteomics, western blot to validate potential biomarkers and development of diagnostic kit for early pregnancy diagnosis in cows.

Keywords: bRPLC, Cluego, ESI, iTRAQ, KEGG, Panther

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521 Digital Transformation and Digitalization of Public Administration

Authors: Govind Kumar

Abstract:

The concept of ‘e-governance’ that was brought about by the new wave of reforms, namely ‘LPG’ in the early 1990s, has been enabling governments across the globe to digitally transform themselves. Digital transformation is leading the governments with qualitative decisions, optimization in rational use of resources, facilitation of cost-benefit analyses, and elimination of redundancy and corruption with the help of ICT-based applications interface. ICT-based applications/technologies have enormous potential for impacting positive change in the social lives of the global citizenry. Supercomputers test and analyze millions of drug molecules for developing candidate vaccines to combat the global pandemic. Further, e-commerce portals help distribute and supply household items and medicines, while videoconferencing tools provide a visual interface between the clients and hosts. Besides, crop yields are being maximized with the help of drones and machine learning, whereas satellite data, artificial intelligence, and cloud computing help governments with the detection of illegal mining, tackling deforestation, and managing freshwater resources. Such e-applications have the potential to take governance an extra mile by achieving 5 Es (effective, efficient, easy, empower, and equity) of e-governance and six Rs (reduce, reuse, recycle, recover, redesign and remanufacture) of sustainable development. If such digital transformation gains traction within the government framework, it will replace the traditional administration with the digitalization of public administration. On the other hand, it has brought in a new set of challenges, like the digital divide, e-illiteracy, technological divide, etc., and problems like handling e-waste, technological obsolescence, cyber terrorism, e-fraud, hacking, phishing, etc. before the governments. Therefore, it would be essential to bring in a rightful mixture of technological and humanistic interventions for addressing the above issues. This is on account of the reason that technology lacks an emotional quotient, and the administration does not work like technology. Both are self-effacing unless a blend of technology and a humane face are brought in into the administration. The paper will empirically analyze the significance of the technological framework of digital transformation within the government set up for the digitalization of public administration on the basis of the synthesis of two case studies undertaken from two diverse fields of administration and present a future framework of the study.

Keywords: digital transformation, electronic governance, public administration, knowledge framework

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520 Modernization of Translation Studies Curriculum at Higher Education Level in Armenia

Authors: A. Vahanyan

Abstract:

The paper touches upon the problem of revision and modernization of the current curriculum on translation studies at the Armenian Higher Education Institutions (HEIs). In the contemporary world where quality and speed of services provided are mostly valued, certain higher education centers in Armenia though do not demonstrate enough flexibility in terms of the revision and amendment of courses taught. This issue is present for various curricula at the university level and Translation Studies related curriculum, in particular. Technological innovations that are of great help for translators have been long ago smoothly implemented into the global Translation Industry. According to the European Master's in Translation (EMT) framework, translation service provision comprises linguistic, intercultural, information mining, thematic, and technological competencies. Therefore, to form the competencies mentioned above, the curriculum should be seriously restructured to meet the modern education and job market requirements, relevant courses should be proposed. New courses, in particular, should focus on the formation of technological competences. These suggestions have been made upon the author’s research of the problem across various HEIs in Armenia. The updated curricula should include courses aimed at familiarization with various computer-assisted translation (CAT) tools (MemoQ, Trados, OmegaT, Wordfast, etc.) in the translation process, creation of glossaries and termbases compatible with different platforms), which will ensure consistency in translation of similar texts and speeding up the translation process itself. Another aspect that may be strengthened via curriculum modification is the introduction of interdisciplinary and Project-Based Learning courses, which will enable info mining and thematic competences, which are of great importance as well. Of course, the amendment of the existing curriculum with the mentioned courses will require corresponding faculty development via training, workshops, and seminars. Finally, the provision of extensive internship with translation agencies is strongly recommended as it will ensure the synthesis of theoretical background and practical skills highly required for the specific area. Summing up, restructuring and modernization of the existing curricula on Translation Studies should focus on three major aspects, i.e., introduction of new courses that meet the global quality standards of education, professional development for faculty, and integration of extensive internship supervised by experts in the field.

Keywords: competencies, curriculum, modernization, technical literacy, translation studies

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