Search results for: complete genome sequencing
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3078

Search results for: complete genome sequencing

2448 Synergism in the Inquiry Lab: An Analysis of Time Targets and Achievement

Authors: John M. Basey, Clinton D. Francis, Maxwell B. Joseph

Abstract:

After gathering data from experimental procedures, inquiry-oriented-science labs often allow students the freedom to stay and complete the write up in class or leave lab early and complete the write up later. Teachers must decide whether to allow students this freedom to self-regulate this time. Student interviews have indicated four time-target strategies that may influence how students utilize this time: grade-target-A, grade-target-C, time-limited, and proficiency. The hypothesis tested was that variability in class composition relative to the four grade-target strategies has an impact on when students leave class, which in turn may influence their overall learning as exemplified by grades. Students were divided into the four indicated groups with a survey. Class composition and the GTA teaching the class had significant impacts on how long students stayed in class with class composition having the greatest impact. A factor analysis identified two factors. Factor 1 included classes with percentages of grade-target students opposite time-limited/proficiency students and explained 43% of the variance. Factor 2 included classes with percentages of grade-target-A/proficiency students opposite grade-target-C students and explained 33% of the variance. Students who stayed longer received significantly higher grades (P = 0.008) with no significant relationships between grade and Factor 1 or Factor 2 (P > 0.05). The time students stayed in class was significantly positively related to Factor 1 (P = 0.006) and significantly negatively related to Factor 2 (P = 0.008). These results support the hypothesis and indicate that teachers may want to know the composition of student-target strategies before deciding on how to have students allocate study time at the end of inquiry-oriented labs. According to these results, ideal classes for self-regulation have a high proportion of proficiency and time-limited students and a low proportion of grade-target students, or a high proportion of grade-target-A and proficiency students and a low proportion of grade-target-C students. Non-ideal classes for self-regulation were comprised of the inverse proportions.

Keywords: grades, inquiry lab design, synergism in student motivation, class composition

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2447 Allelic Diversity of Productive, Reproductive and Fertility Traits Genes of Buffalo and Cattle

Authors: M. Moaeen-ud-Din, G. Bilal, M. Yaqoob

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Identification of genes of importance regarding production traits in buffalo is impaired by a paucity of genomic resources. Choice to fill this gap is to exploit data available for cow. The cross-species application of comparative genomics tools is potential gear to investigate the buffalo genome. However, this is dependent on nucleotide sequences similarity. In this study gene diversity between buffalo and cattle was determined by using 86 gene orthologues. There was about 3% difference in all genes in term of nucleotide diversity; and 0.267±0.134 in amino acids indicating the possibility for successfully using cross-species strategies for genomic studies. There were significantly higher non synonymous substitutions both in cattle and buffalo however, there was similar difference in term of dN – dS (4.414 vs 4.745) in buffalo and cattle respectively. Higher rate of non-synonymous substitutions at similar level in buffalo and cattle indicated a similar positive selection pressure. Results for relative rate test were assessed with the chi-squared test. There was no significance difference on unique mutations between cattle and buffalo lineages at synonymous sites. However, there was a significance difference on unique mutations for non synonymous sites indicating ongoing mutagenic process that generates substitutional mutation at approximately the same rate at silent sites. Moreover, despite of common ancestry, our results indicate a different divergent time among genes of cattle and buffalo. This is the first demonstration that variable rates of molecular evolution may be present within the family Bovidae.

Keywords: buffalo, cattle, gene diversity, molecular evolution

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2446 Modern Proteomics and the Application of Machine Learning Analyses in Proteomic Studies of Chronic Kidney Disease of Unknown Etiology

Authors: Dulanjali Ranasinghe, Isuru Supasan, Kaushalya Premachandra, Ranjan Dissanayake, Ajith Rajapaksha, Eustace Fernando

Abstract:

Proteomics studies of organisms are considered to be significantly information-rich compared to their genomic counterparts because proteomes of organisms represent the expressed state of all proteins of an organism at a given time. In modern top-down and bottom-up proteomics workflows, the primary analysis methods employed are gel–based methods such as two-dimensional (2D) electrophoresis and mass spectrometry based methods. Machine learning (ML) and artificial intelligence (AI) have been used increasingly in modern biological data analyses. In particular, the fields of genomics, DNA sequencing, and bioinformatics have seen an incremental trend in the usage of ML and AI techniques in recent years. The use of aforesaid techniques in the field of proteomics studies is only beginning to be materialised now. Although there is a wealth of information available in the scientific literature pertaining to proteomics workflows, no comprehensive review addresses various aspects of the combined use of proteomics and machine learning. The objective of this review is to provide a comprehensive outlook on the application of machine learning into the known proteomics workflows in order to extract more meaningful information that could be useful in a plethora of applications such as medicine, agriculture, and biotechnology.

Keywords: proteomics, machine learning, gel-based proteomics, mass spectrometry

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2445 Evaluation of Hollocelulase Production for Lignocellulosic Biomass Degradation by Penicillium polonicum

Authors: H. M. Takematsu, B. R. De Camargo, E. F. Noronha

Abstract:

The use of hydrolyzing enzymes for degradation of lignocellulosic biomass is of great concern for the production of second generation ethanol. Although many hollocelulases have already been described in the literature, much more has to be discovered. Therefore, the aim of this study to evaluate hollocelulase production of P. polonicum grown in liquid media containing sugarcane bagasse as the carbon source. From a collection of twenty fungi isolated from Cerrado biome soil, P. polonicum was molecular identified by sequencing of ITS4, βtubulin and Calmodulin genes, and has been chosen to be further investigated regarding its potential production of hydrolyzing enzymes. Spore suspension (1x10-6 ml-1) solution was inoculated in sterilized minimal liquid medium containing 0,5%(w/v) of non-pretreated sugarcane bagasse as the carbon source, and incubated in shaker incubator at 28°C and 120 rpm. The supernatant obtained, was subjected to enzymatic assays to analyze xylanase, mannanase, pectinase and endoglucanase activities. Xylanase activity showed better results (67,36 UI/mg). Xylanases bands were indicated by zymogram and SDS-PAGE, and one of them was partially purified and characterized. It showed maximum activity at 50 °C, was thermostable for 72h at 40°C, and pH5.0 was the optimum observed. This study presents P. polonicum as an interesting source of hollocelulases, especially xylanase, for lignocellulose bio-conversion processes with commercial use.

Keywords: sugarcane bagasse, Cerrado biome , hollocelulase, lignocellulosic biomass

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2444 Wall Shear Stress Under an Impinging Planar Jet Using the Razor Blade Technique

Authors: A. Ritcey, J. R. Mcdermid, S. Ziada

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Wall shear stress was experimentally measured under a planar impinging air jet as a function of jet Reynolds number (Rejet = 5000, 8000, 11000) and different normalized impingement distances (H/D = 4, 6, 8, 10, 12) using the razor blade technique to complete a parametric study. The wall pressure, wall pressure gradient, and wall shear stress information were obtained.

Keywords: experimental fluid mechanics, impinging planar jets, skin friction factor, wall shear stress

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2443 Field Study of Chlorinated Aliphatic Hydrocarbons Degradation in Contaminated Groundwater via Micron Zero-Valent Iron Coupled with Biostimulation

Authors: Naijin Wu, Peizhong Li, Haijian Wang, Wenxia Wei, Yun Song

Abstract:

Chlorinated aliphatic hydrocarbons (CAHs) pollution poses a severe threat to human health and is persistent in groundwater. Although chemical reduction or bioremediation is effective, it is still hard to achieve their complete and rapid dechlorination. Recently, the combination of zero-valent iron and biostimulation has been considered to be one of the most promising strategies, but field studies of this technology are scarce. In a typical site contaminated by various types of CAHs, basic physicochemical parameters of groundwater, CAHs and their product concentrations, and microbial abundance and diversity were monitored after a remediation slurry containing both micron zero-valent iron (mZVI) and biostimulation components were directly injected into the aquifer. Results showed that groundwater could form and keep low oxidation-reduction potential (ORP), a neutral pH, and anoxic conditions after different degrees of fluctuations, which was benefit for the reductive dechlorination of CAHs. The injection also caused an obvious increase in the total organic carbon (TOC) concentration and sulfate reduction. After 253 days post-injection, the mean concentration of total chlorinated ethylene (CEE) from two monitoring wells decreased from 304 μg/L to 8 μg/L, and total chlorinated ethane (CEA) decreased from 548 μg/L to 108 μg/L. Occurrence of chloroethane (CA) suggested that hydrogenolysis dechlorination was one of the main degradation pathways for CEA, and also hints that biological dechlorination was activated. A significant increase of ethylene at day 67 post-injection indicated that dechlorination was complete. Additionally, the total bacterial counts increased by 2-3 orders of magnitude after 253 days post-injection. And the microbial species richness decreased and gradually changed to anaerobic/fermentative bacteria. The relative abundance of potential degradation bacteria increased corresponding to the degradation of CAHs. This work demonstrates that mZVI and biostimulation can be combined to achieve the efficient removal of various CAHs from contaminated groundwater sources.

Keywords: chlorinated aliphatic hydrocarbons, groundwater, field study, zero-valent iron, biostimulation

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2442 Framework Proposal on How to Use Game-Based Learning, Collaboration and Design Challenges to Teach Mechatronics

Authors: Michael Wendland

Abstract:

This paper presents a framework to teach a methodical design approach by the help of using a mixture of game-based learning, design challenges and competitions as forms of direct assessment. In today’s world, developing products is more complex than ever. Conflicting goals of product cost and quality with limited time as well as post-pandemic part shortages increase the difficulty. Common design approaches for mechatronic products mitigate some of these effects by helping the users with their methodical framework. Due to the inherent complexity of these products, the number of involved resources and the comprehensive design processes, students very rarely have enough time or motivation to experience a complete approach in one semester course. But, for students to be successful in the industrial world, it is crucial to know these methodical frameworks and to gain first-hand experience. Therefore, it is necessary to teach these design approaches in a real-world setting and keep the motivation high as well as learning to manage upcoming problems. This is achieved by using a game-based approach and a set of design challenges that are given to the students. In order to mimic industrial collaboration, they work in teams of up to six participants and are given the main development target to design a remote-controlled robot that can manipulate a specified object. By setting this clear goal without a given solution path, a constricted time-frame and limited maximal cost, the students are subjected to similar boundary conditions as in the real world. They must follow the methodical approach steps by specifying requirements, conceptualizing their ideas, drafting, designing, manufacturing and building a prototype using rapid prototyping. At the end of the course, the prototypes will be entered into a contest against the other teams. The complete design process is accompanied by theoretical input via lectures which is immediately transferred by the students to their own design problem in practical sessions. To increase motivation in these sessions, a playful learning approach has been chosen, i.e. designing the first concepts is supported by using lego construction kits. After each challenge, mandatory online quizzes help to deepen the acquired knowledge of the students and badges are awarded to those who complete a quiz, resulting in higher motivation and a level-up on a fictional leaderboard. The final contest is held in presence and involves all teams with their functional prototypes that now need to contest against each other. Prices for the best mechanical design, the most innovative approach and for the winner of the robotic contest are awarded. Each robot design gets evaluated with regards to the specified requirements and partial grades are derived from the results. This paper concludes with a critical review of the proposed framework, the game-based approach for the designed prototypes, the reality of the boundary conditions, the problems that occurred during the design and manufacturing process, the experiences and feedback of the students and the effectiveness of their collaboration as well as a discussion of the potential transfer to other educational areas.

Keywords: design challenges, game-based learning, playful learning, methodical framework, mechatronics, student assessment, constructive alignment

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2441 Identification of Cellulose-Hydrolytic Thermophiles Isolated from Sg. Klah Hot Spring Based on 16S rDNA Gene Sequence

Authors: M. J. Norashirene, Y. Zakiah, S. Nurdiana, I. Nur Hilwani, M. H. Siti Khairiyah, M. J. Muhamad Arif

Abstract:

In this study, six bacterial isolates of a slightly thermophilic organism from the Sg. Klah hot spring, Malaysia were successfully isolated and designated as M7T55D1, M7T55D2, M7T55D3, M7T53D1, M7T53D2 and M7T53D3 respectively. The bacterial isolates were screened for their cellulose hydrolytic ability on Carboxymethlycellulose agar medium. The isolated bacterial strains were identified morphologically, biochemically and molecularly with the aid of 16S rDNA sequencing. All of the bacteria showed their optimum growth at a slightly alkaline pH of 7.5 with a temperature of 55°C. All strains were Gram-negative, non-spore forming type, strictly aerobic, catalase-positive and oxidase-positive with the ability to produce thermostable cellulase. Based on BLASTn results, bacterial isolates of M7T55D2 and M7T53D1 gave the highest homology (97%) with similarity to Tepidimonas ignava while isolates M7T55D1, M7T55D3, M7T53D2 and M7T53D3 showed their closest homology (97%-98%) with Tepidimonas thermarum. These cellulolytic thermophiles might have a commercial potential to produce valuable thermostable cellulase.

Keywords: cellulase, cellulolytic, thermophiles, 16S rDNA gene

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2440 Molecular Detection and Isolation of Benzimidazole Resistant Haemonchus contortus from Pakistan

Authors: K. Ali, M. F. Qamar, M. A. Zaman, M. Younus, I. Khan, S. Ehtisham-ul-Haque, R. Tamkeen, M. I. Rashid, Q. Ali

Abstract:

This study centers on molecular identification of Haemonchus contortus and isolation of Benz-imidazoles (BZ) resistant strains. Different abattoirs’ of two geographic regions of Punjab (Pakistan) were frequently visited for the collection of worms. Out of 1500 (n=1500) samples that were morphologically confirmed as H. contortus, 30 worms were subjected to molecular procedures for isolation of resistant strains. Resistant worms (n=8) were further subjected to DNA gene sequencing. Bio edit sequence alignment editor software was used to detect the possible mutation, deletion, replacement of nucleotides. Genetic diversity was noticed and genetic variation existing in β-tubulin isotype 1 of the H. contortus population of small ruminants of different regions considered in this study. H. contortus showed three different type of genetic sequences. 75%, 37.5%, 25% and 12.5% of the studied samples showed 100% query cover and identity with isolates and clones of China, UK, Australia and other countries, respectively. Interestingly the neighbor countries such as India and Iran haven’t many similarities with the Pakistani isolates. Thus, it suggests that population density of same genetic makeup H. contortus is scattered worldwide rather than clustering in a single region.

Keywords: Haemonchus contortus, Benzimidazole resistant, β-tubulin-1 gene, abattoirs

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2439 Correlation of P53 Gene Expression With Serum Alanine Transaminase Levels and Hepatitis B Viral Load in Cirrhosis and Hepatocellular Carcinoma Patients

Authors: Umme Shahera, Saifullah Munshi, Munira Jahan, Afzalun Nessa, Shahinul Alam, Shahina Tabassum

Abstract:

The development of HCC is a multi-stage process. Several extrinsic factors, such as aflatoxin, HBV, nutrition, alcohol, and trace elements are thought to initiate or/and promote the hepatocarcinogenesis. Alteration of p53 status is an important intrinsic factor in this process as p53 is essential for preventing inappropriate cell proliferation and maintaining genome integrity following genotoxic stress. This study was designed to assess the correlation of p53 gene expression with HBV-DNA and serum Alanine transaminase (ALT) in patients with cirrhosis and HCC. The study was conducted among 60 patients. The study population were divided into four groups (15 in each groups)-HBV positive cirrhosis, HBV negative cirrhosis, HBV positive HCC and HBV negative HCC. Expression of p53 gene was observed using real time PCR. P53 gene expressions in the above mentioned groups were correlated with serum ALT level and HBV viral load. p53 gene was significantly higher in HBV-positive patients with HCC than HBV-positive cirrhosis. Similarly, the expression of p53 was significantly higher in HBV-positive HCC than HBV-negative HCC patients. However, the expression of p53 was reduced in HBV-positive cirrhosis in comparison with HBV-negative cirrhosis. P53 gene expression in liver was not correlated with the serum levels of ALT in any of the study groups. HBV- DNA load also did not correlated with p53 gene expression in HBV positive HCC and HBV positive cirrhosis patients. This study shows that there was no significant change with the expression of p53 gene in any of the study groups with ALT level or viral load, though differential expression of p53 gene were observed in cirrhosis and HCC patients.

Keywords: P53, ALT, HBV-DNA, liver cirrhosis, hepatocellular carcinoma

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2438 Region-Specific Secretory Protein, α2M, in Male Reproductive Tract of the Blue Crab And Its Dynamics during Sperm transit towards Female Spermatheca

Authors: Thanyaporn Senarai, Rapeepun Vanichviriyakit, Shinji Miyata, Chihiro Sato, Prapee Sretarugsa, Wattana Weerachatyanukul, Ken Kitajima

Abstract:

In this study, we characterized a region-specific 250 kDa protein that was secreted of MSD fluid, which is believed to play dual functions in forming a spermatophoric wall for sperm physical protection, and in sperm membrane modification as part of sperm maturation process. The partial amino acid sequence and N-terminal sequencing revealed that the MSD-specific 250 kDa protein showed a high similarity with a plasma-rich protein, α-2 macroglobulin (α2M), so termed pp-α2M. This protein was a large glycoprotein contained predominantly mannose and GlcNAc. The expression of pp-α2M mRNA was detected in spermatic duct (SD), androgenic gland (AG) and hematopoietic tissue, while the protein expression was rather specific to the apical cytoplasm of MSD epithelium. The secretory pp-α2M in MSD fluid was acquired onto the MSD sperm membrane and was also found within the matrix of the acrosome. Distally, pp-α2M was removed from spermathecal sperm membrane, while its level kept constant in the sperm AC. Together the results indicate that pp-α2M is a 250 kDa region-specific secretory protein which plays roles in sperm physical protection and also acts as maturation factor in the P. pelagicus sperm.

Keywords: alpha-2 macroglobulin, blue swimming crab, sperm maturation, spermatic duct

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2437 Comparison of 18F-FDG and 11C-Methionine PET-CT for Assessment of Response to Neoadjuvant Chemotherapy in Locally Advanced Breast Carcinoma

Authors: Sonia Mahajan Dinesh, Anant Dinesh, Madhavi Tripathi, Vinod Kumar Ramteke, Rajnish Sharma, Anupam Mondal

Abstract:

Background: Neo-adjuvant chemotherapy plays an important role in treatment of breast cancer by decreasing the tumour load and it offers an opportunity to evaluate response of primary tumour to chemotherapy. Standard anatomical imaging modalities are unable to accurately reflect the response to chemotherapy until several cycles of drug treatment have been completed. Metabolic imaging using tracers like 18F-fluorodeoxyglucose (FDG) as a marker of glucose metabolism or amino acid tracers like L-methyl-11C methionine (MET) have potential role for the measurement of treatment response. In this study, our objective was to compare these two PET tracers for assessment of response to neoadjuvant chemotherapy, in locally advanced breast carcinoma. Methods: In our prospective study, 20 female patients with histology proven locally advanced breast carcinoma underwent PET-CT imaging using FDG and MET before and after three cycles of neoadjuvant chemotherapy (CAF regimen). Thereafter, all patients were taken for MRM and the resected specimen was sent for histo-pathological analysis. Tumour response to the neoadjuvant chemotherapy was evaluated by PET-CT imaging using PERCIST criteria and correlated with histological results. Responses calculated were compared for statistical significance using paired t- test. Results: Mean SUVmax for primary lesion in FDG PET and MET PET was 15.88±11.12 and 5.01±2.14 respectively (p<0.001) and for axillary lymph nodes was 7.61±7.31 and 2.75±2.27 respectively (p=0.001). Statistically significant response in primary tumour and axilla was noted on both FDG and MET PET after three cycles of NAC. Complete response in primary tumour was seen in only 1 patient in FDG and 7 patients in MET PET (p=0.001) whereas there was no histological complete resolution of tumor in any patient. Response to therapy in axillary nodes noted on both PET scans were similar (p=0.45) and correlated well with histological findings. Conclusions: For the primary breast tumour, FDG PET has a higher sensitivity and accuracy than MET PET and for axilla both have comparable sensitivity and specificity. FDG PET shows higher target to background ratios so response is better predicted for primary breast tumour and axilla. Also, FDG-PET is widely available and has the advantage of a whole body evaluation in one study.

Keywords: 11C-methionine, 18F-FDG, breast carcinoma, neoadjuvant chemotherapy

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2436 Wind Speed Data Analysis in Colombia in 2013 and 2015

Authors: Harold P. Villota, Alejandro Osorio B.

Abstract:

The energy meteorology is an area for study energy complementarity and the use of renewable sources in interconnected systems. Due to diversify the energy matrix in Colombia with wind sources, is necessary to know the data bases about this one. However, the time series given by 260 automatic weather stations have empty, and no apply data, so the purpose is to fill the time series selecting two years to characterize, impute and use like base to complete the data between 2005 and 2020.

Keywords: complementarity, wind speed, renewable, colombia, characteri, characterization, imputation

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2435 Design and in Slico Study of the Truncated Spike-M-N SARS-CoV-2 as a Novel Effective Vaccine Candidate

Authors: Aghasadeghi MR., Bahramali G., Sadat SM., Sadeghi SA., Yousefi M., Khodaei K., Ghorbani M., Sadat Larijani M.

Abstract:

Background:The emerging COVID-19 pandemic is a serious concernfor the public health worldwide. Despite the many mutations in the virus genome, it is important to find an effective vaccine against viral mutations. Therefore, in current study, we aimed at immunoinformatic evaluation of the virus proteins immunogenicity to design a preventive vaccine candidate, which could elicit humoral and cellular immune responses as well. Methods:Three antigenic regions are included;Spike, Membrane, and Nucleocapsid amino acid sequences were obtained, and possible fusion proteins were assessed andcompared by immunogenicity, structural features, and population coverage. The best fusion protein was also evaluated for MHC-I and MHC-II T-cell epitopes and the linear and conformational B-cell epitopes. Results: Among the four predicted models, the truncated Spike protein in fusion with M and N proteins is composed of 24 highly immunogenic human MHC class I and 29 MHC class II, along with 14 B-cell linear and 61 discontinues epitopes. Also, the selected protein has high antigenicity and acceptable population coverage of 82.95% in Iran and 92.51% in Europe. Conclusion: The data indicate that the truncated Spike-M-N SARS-CoV-2form which could be potential targets of neutralizing antibodies. The protein also has the ability to stimulate humoral and cellular immunity. The in silico study provided the fusion protein as a potential preventive vaccine candidate for further in vivo evaluation.

Keywords: SARS-CoV-2, immunoinformatic, protein, vaccine

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2434 Effects of Twitter Interactions on Self-Esteem and Narcissistic Behaviour

Authors: Leena-Maria Alyedreessy

Abstract:

Self-esteem is thought to be determined by ones’ own feeling of being included, liked and accepted by others. This research explores whether this concept may also be applied in the virtual world and assesses whether there is any relationship between Twitter users' self-esteem and the amount of interactions they receive. 20 female Arab participants were given a survey asking them about their Twitter interactions and their feelings of having an imagined audience to fill out and a Rosenberg Self-Esteem Assessment to complete. After completion and statistical analysis, results showed a significant correlation between the feeling of being Twitter elite, the feeling of having a lot of people listening to your tweets and having a lot of interactions with high self-esteem. However, no correlations were detected for low-self-esteem and low interactions.

Keywords: twitter, social media, self-esteem, narcissism, interactions

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2433 Carbohydrates Quantification from Agro-Industrial Waste and Fermentation with Lactic Acid Bacteria

Authors: Prittesh Patel, Bhavika Patel, Ramar Krishnamurthy

Abstract:

Present study was conducted to isolate lactic acid bacteria (LAB) from Oreochromis niloticus and Nemipterus japonicus fish gut. The LAB isolated were confirmed through 16s rRNA sequencing. It was observed that isolated Lactococcus spp. were able to tolerate NaCl and bile acid up to certain range. The isolated Lactococcus spp. were also able to survive in acidic and alkaline conditions. Further agro-industrial waste like peels of pineapple, orange, lemon, sugarcane, pomegranate; sweet lemon was analyzed for their polysaccharide contents and prebiotic properties. In the present study, orange peels, sweet lemon peels, and pineapple peels give maximum indigestible polysaccharide. To evaluate synbiotic effect combination of probiotic and prebiotic were analyzed under in vitro conditions. Isolates Lactococcus garvieae R3 and Lactococcus sp. R4 reported to have better fermentation efficiency with orange, sweet lemon and pineapple compare to lemon, sugarcane and pomegranate. The different agro-industrial waste evaluated in this research resulted in being a cheap and fermentable carbon source by LAB.

Keywords: agro-industrial waste, lactic acid bacteria, prebiotic, probiotic, synbiotic

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2432 Characterization of Enterotoxigenic Escherichia coli CS6 Promoter

Authors: Mondal Indranil, Bhakat Debjyoti, Mukhopadayay Asish K., Chatterjee Nabendu S.

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CS6 is the prevalent CF in our region and deciphering its molecular regulators would play a pivotal role in reducing the burden of ETEC pathogenesis. In prokaryotes, most of the genes are under the control of one operon and the promoter present upstream of the gene regulates the transcription of that gene. Here the promoter of CS6 was characterized by computational method and further analyzed by β-galactosidase assay and sequencing. Promoter constructs and deletions were prepared as required to analyze promoter activity. The effect of different additives on the CS6 promoter was analysed by the β-galactosidase assay. Bioinformatics analysis done by Softberry/BPROM predicted fur, lrp, and crp boxes, -10 and -35 region upstream of the CS6 gene. The promoter construction in no promoter plasmid pTL61T showed that region -573 to +1 is actually the promoter region as predicted. Sequential deletion of the region upstream of CS6 revealed that promoter activity remains the same when -573bp to -350bp is deleted. But after the deletion of the upstream region -350 bp to -255bp, promoter expression decreases drastically to 26%. Further deletion also decreases promoter activity up to a little range. So the region -355bp to -255bp holds the promoter sequence for the CS6 gene. Additives like iron, NaCl, etc., modulate promoter activity in a dose-dependent manner. From the promoter analysis, it can be said that the minimum region lies between -254 and +1. Important region(s) lies between -350 bp to -255 bp upstream in the promoter, which might have important elements needed to control CS6 gene expression.

Keywords: microbiology, promoter, colonization factor, ETEC

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2431 Anaerobic Digestion of Coffee Wastewater from a Fast Inoculum Adaptation Stage: Replacement of Complex Substrate

Authors: D. Lepe-Cervantes, E. Leon-Becerril, J. Gomez-Romero, O. Garcia-Depraect, A. Lopez-Lopez

Abstract:

In this study, raw coffee wastewater (CWW) was used as a complex substrate for anaerobic digestion. The inoculum adaptation stage, microbial diversity analysis and biomethane potential (BMP) tests were performed. A fast inoculum adaptation stage was used by the replacement of vinasse to CWW in an anaerobic sequential batch reactor (AnSBR) operated at mesophilic conditions. Illumina MiSeq sequencing was used to analyze the microbial diversity. While, BMP tests using inoculum adapted to CWW were carried out at different inoculum to substrate (I/S) ratios (2:1, 3:1 and 4:1, on a VS basis). Results show that the adaptability percentage was increased gradually until it reaches the highest theoretical value in a short time of 10 d; with a methane yield of 359.10 NmL CH4/g COD-removed; Methanobacterium beijingense was the most abundant microbial (75%) and the greatest specific methane production was achieved at I/S ratio 4:1, whereas the lowest was obtained at 2:1, with BMP values of 320 NmL CH4/g VS and 151 NmL CH4/g VS, respectively. In conclusion, gradual replacement of substrate was a feasible method to adapt the inoculum in a short time even using complex raw substrates, whereas in the BMP tests, the specific methane production was proportional to the initial amount of inoculum.

Keywords: anaerobic digestion, biomethane potential test, coffee wastewater, fast inoculum adaptation

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2430 Biodegradation Study of Diethyl Phthalate Using Bacteria Isolated from Plastic Industry Wastewater Discharge Site

Authors: Sangram Shamrao Patil, Hara Mohan Jena

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Phthalates are among the most common organic pollutant since they have become widespread in the environment and found in sediments, natural waters, soils, plants, landfill leachates, biota including human tissue and aquatic organisms. Diethyl phthalate (DEP) is a low molecular weight phthalate which has wide applications as plasticizer and become a major cause of environmental pollution. Environmental protection agency (EPA) listed DEP as priority pollutant because of its toxicity and they recommended human health ambient water quality criterion for diethyl phthalate (DEP) as 4 mg/l. Therefore, wastes containing phthalates require proper treatment before being discharged into the environment. Biodegradation is attractive and efficient treatment method as it is cost effective and produces non-toxic end products. In the present study, a DEP degrading aerobic bacterium was isolated from soil contaminated with plastic industry wastewater. Morphological and biochemical characteristics of isolate were performed. 16S rRNA sequencing and phylogenetic analysis of isolate was carried out and it was identified as Empedobacter brevis. Isolate has been found to tolerate up to 1650 mg/l of DEP. This study will be significant for exploring an application of microbes for remediation of phthalates and development of a suitable bioreactor.

Keywords: diethyl phthalate, plasticizer, pollutant, biodegradation

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2429 Geriatric Nutrition: An Important Marker for Oral Health Related Quality of Life of Complete Denture Patients

Authors: Rajlakshmi Banerjee, Sujoy Banerjee

Abstract:

The prevalence of malnutrition, as the most common cause of morbidity and mortality increases with old age, especially in developing countries like India, because of many factors out of which dentate status is one. It affects diet, nutritional status and general health due to reduced chewing ability and poor food choices. Aim and Objective: This study aimed to evaluate the effect of nutritional status, dietary intake on the Oral Health-related Quality of life (OHRQoL) of elderly edentulous complete denture wearing patients and to know whether they are at a higher risk of malnutrition. The objective was to assess the need to include dietary and nutritional counselling during Prosthodontic rehabilitation of elderly edentulous patients. Materials and method: A cross-sectional study was conducted among 200 elderly denture wearing patients above 60 yrs of age from Nagpur, Maharashtra. The majority of study subjects (60%) were between the age group 60 and 75 years. Mini-Nutritional Assessment (MNA) Questionnaire was used to assess nutritional status and General Oral Health Assessment Index (GOHAI) questionnaire was used to determine the Oral health related Quality of life of these patients.Descriptive statistics was used to analyze data using SPSS version 21. Results: Among the assessed subjects, nearly 80% of them had total scores of GOHAI between 12 and 57 which require ‘needed dental care.’ As per MNA, 10.5% had adequate nutrition, 70 % were at risk of malnutrition, and remaining 19.5 % of subjects were malnourished. There was a significant correlation between GOHAI and MNA scores. A strong association was found between mean GOHAI and MNA scores and thereby nutritional status and OHRQOL.Conclusion: The use of conventional dentures increases the risk of malnutrition in the elderly due to inability to eat and chew food properly and therefore severely affecting the quality of life. Dietary analysis and counselling should be strictly incorporated into geriatric treatment planning during Prosthetic rehabilitation.

Keywords: general oral health assessment index, General Oral Health Assessment Index (GOHAI), nutritional assessment, mini-nutritional assessment, Mini-Nutritional Assessment (MNA), quality of life

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2428 Genomic Sequence Representation Learning: An Analysis of K-Mer Vector Embedding Dimensionality

Authors: James Jr. Mashiyane, Risuna Nkolele, Stephanie J. Müller, Gciniwe S. Dlamini, Rebone L. Meraba, Darlington S. Mapiye

Abstract:

When performing language tasks in natural language processing (NLP), the dimensionality of word embeddings is chosen either ad-hoc or is calculated by optimizing the Pairwise Inner Product (PIP) loss. The PIP loss is a metric that measures the dissimilarity between word embeddings, and it is obtained through matrix perturbation theory by utilizing the unitary invariance of word embeddings. Unlike in natural language, in genomics, especially in genome sequence processing, unlike in natural language processing, there is no notion of a “word,” but rather, there are sequence substrings of length k called k-mers. K-mers sizes matter, and they vary depending on the goal of the task at hand. The dimensionality of word embeddings in NLP has been studied using the matrix perturbation theory and the PIP loss. In this paper, the sufficiency and reliability of applying word-embedding algorithms to various genomic sequence datasets are investigated to understand the relationship between the k-mer size and their embedding dimension. This is completed by studying the scaling capability of three embedding algorithms, namely Latent Semantic analysis (LSA), Word2Vec, and Global Vectors (GloVe), with respect to the k-mer size. Utilising the PIP loss as a metric to train embeddings on different datasets, we also show that Word2Vec outperforms LSA and GloVe in accurate computing embeddings as both the k-mer size and vocabulary increase. Finally, the shortcomings of natural language processing embedding algorithms in performing genomic tasks are discussed.

Keywords: word embeddings, k-mer embedding, dimensionality reduction

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2427 Functional Expression and Characterization of a Novel Indigenous Endo-Beta 1,4- Glucanase from Apis mellifera

Authors: Amtul Jamil Sami

Abstract:

Apis mellifera is an insect of immense economic importance lives on rich carbohydrate diet including cellulose, nectar, honey and pollen. The carbohydrate metabolism in A mellifera has not been understood fully, as there are no data available, on the functional expression of cellulase gene. The cellulose hydrolyzing enzyme is required for the digestion of pollen cellulose wall, to release the important nutrients (amino acids, minerals, vitamins etc.) from the pollen. A dissection of Apis genome had revealed that there is one gene present for the expression of endo-beta-1,4-glucanase, for cellulose hydrolysis. In the presented work, functional expression of endo-beta-1,4 glucanase gene is reported. Total soluble proteins of the honey bee were isolated and were tested cellulose hydrolyzing enzyme activity, using carboxy-methyl cellulose, as a substrate. A mellifera proteins were able to hydrolyze carboxy-methyl cellulose, confirming its endo- type mode of action. Endo beta-1,4 glucanase enzyme was only present in the gut tissues, no activity was detected in the salivary glands. The pH optima of the enzyme were in the acidic pH range of 4-5-5-0, indicating its metabolic role in the acidic stomach of A mellifera. The reported enzyme is unique, as endo-beta- 1,4 glucanase was able to generate non reducing sugar, as an end product. The results presented, are supportive to the information that the honey bee is capable of producing its novel endo-beta-1,4 glucanase. Further it could be helpful, in understanding, the carbohydrate metabolism in A mellifera.

Keywords: honey bees, Endo-beta 1, 4- glucanase, Apis mellifera, functional expression

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2426 Influence Zone of Strip Footing on Untreated and Cement Treated Sand Mat Underlain by Soft Clay (2nd reviewed)

Authors: Sharifullah Ahmed

Abstract:

Shallow foundation on soft soils without ground improvement can represent a high level of settlement. In such a case, an alternative to pile foundations may be shallow strip footings placed on a soil system in which the upper layer is untreated or cement-treated compacted sand to limit the settlement within a permissible level. This research work deals with a rigid plane-strain strip footing of 2.5m width placed on a soil consisting of untreated or cement treated sand layer underlain by homogeneous soft clay. Both the thin and thick compared the footing width was considered. The soft inorganic cohesive NC clay layer is considered undrained for plastic loading stages and drained in consolidation stages, and the sand layer is drained in all loading stages. FEM analysis was done using PLAXIS 2D Version 8.0 with a model consisting of clay deposits of 15m thickness and 18m width. The soft clay layer was modeled using the Hardening Soil Model, Soft Soil Model, Soft Soil Creep model, and the upper improvement layer was modeled using only the Hardening Soil Model. The system is considered fully saturated. The value of natural void ratio 1.2 is used. Total displacement fields of strip footing and subsoil layers in the case of Untreated and Cement treated Sand as Upper layer are presented. For Hi/B =0.6 or above, the distribution of major deformation within an upper layer and the influence zone of footing is limited in an upper layer which indicates the complete effectiveness of the upper layer in bearing the foundation effectively in case of the untreated upper layer. For Hi/B =0.3 or above, the distribution of major deformation occurred within an upper layer, and the function of footing is limited in the upper layer. This indicates the complete effectiveness of the cement-treated upper layer. Brittle behavior of cemented sand and fracture or cracks is not considered in this analysis.

Keywords: displacement, ground improvement, influence depth, PLAXIS 2D, primary and secondary settlement, sand mat, soft clay

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2425 Population Structure Analysis of Pakistani Indigenous Cattle Population by Using High Density SNP Array

Authors: Hamid Mustafa, Huson J. Heather, Kim Eiusoo, McClure Matt, Khalid Javed, Talat Nasser Pasha, Afzal Ali1, Adeela Ajmal, Tad Sonstegard

Abstract:

Genetic differences associated with speciation, breed formation or local adaptation can help to preserve and effective utilization of animals in selection programs. Analyses of population structure and breed diversity have provided insight into the origin and evolution of cattle. In this study, we used a high-density panel of SNP markers to examine population structure and diversity among ten Pakistani indigenous cattle breeds. In total, 25 individuals from three cattle populations, including Achi (n=08), Bhagnari (n=04) and Cholistani (n=13) were genotyped for 777, 962 single nucleotide polymorphism (SNP) markers. Population structure was examined using the linkage model in the program STRUCTURE. After characterizing SNP polymorphism in the different populations, we performed a detailed analysis of genetic structure at both the individual and population levels. The whole-genome SNP panel identified several levels of population substructure in the set of examined cattle breeds. We further searched for spatial patterns of genetic diversity among these breeds under the recently developed spatial principal component analysis framework. Overall, such high throughput genotyping data confirmed a clear partitioning of the cattle genetic diversity into distinct breeds. The resulting complex historical origins associated with both natural and artificial selection have led to the differentiation of numerous different cattle breeds displaying a broad phenotypic variety over a short period of time.

Keywords: Pakistan, cattle, genetic diversity, population structure

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2424 A Novel Machine Learning Approach to Aid Agrammatism in Non-fluent Aphasia

Authors: Rohan Bhasin

Abstract:

Agrammatism in non-fluent Aphasia Cases can be defined as a language disorder wherein a patient can only use content words ( nouns, verbs and adjectives ) for communication and their speech is devoid of functional word types like conjunctions and articles, generating speech of with extremely rudimentary grammar . Past approaches involve Speech Therapy of some order with conversation analysis used to analyse pre-therapy speech patterns and qualitative changes in conversational behaviour after therapy. We describe this approach as a novel method to generate functional words (prepositions, articles, ) around content words ( nouns, verbs and adjectives ) using a combination of Natural Language Processing and Deep Learning algorithms. The applications of this approach can be used to assist communication. The approach the paper investigates is : LSTMs or Seq2Seq: A sequence2sequence approach (seq2seq) or LSTM would take in a sequence of inputs and output sequence. This approach needs a significant amount of training data, with each training data containing pairs such as (content words, complete sentence). We generate such data by starting with complete sentences from a text source, removing functional words to get just the content words. However, this approach would require a lot of training data to get a coherent input. The assumptions of this approach is that the content words received in the inputs of both text models are to be preserved, i.e, won't alter after the functional grammar is slotted in. This is a potential limit to cases of severe Agrammatism where such order might not be inherently correct. The applications of this approach can be used to assist communication mild Agrammatism in non-fluent Aphasia Cases. Thus by generating these function words around the content words, we can provide meaningful sentence options to the patient for articulate conversations. Thus our project translates the use case of generating sentences from content-specific words into an assistive technology for non-Fluent Aphasia Patients.

Keywords: aphasia, expressive aphasia, assistive algorithms, neurology, machine learning, natural language processing, language disorder, behaviour disorder, sequence to sequence, LSTM

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2423 The Effect of Subsurface Dam on Saltwater Intrusion in Heterogeneous Coastal Aquifers

Authors: Antoifi Abdoulhalik, Ashraf Ahmed

Abstract:

Saltwater intrusion (SWI) in coastal aquifers has become a growing threat for many countries around the world. While various control measures have been suggested to mitigate SWI, the construction of subsurface physical barriers remains one of the most effective solutions for this problem. In this work, we used laboratory experiments and numerical simulations to investigate the effectiveness of subsurface dams in heterogeneous layered coastal aquifer with different layering patterns. Four different cases were investigated, including a homogeneous (case H), and three heterogeneous cases in which a low permeability (K) layer was set in the top part of the system (case LH), in the middle part of the system (case HLH) and the bottom part of the system (case HL). Automated image analysis technique was implemented to quantify the main SWI parameters under high spatial and temporal resolution. The method also provides transient salt concentration maps, allowing for the first time clear visualization of the spillage of saline water over the dam (advancing wedge condition) as well as the flushing of residual saline water from the freshwater area (receding wedge condition). The SEAWAT code was adopted for the numerical simulations. The results show that the presence of an overlying layer of low permeability enhanced the ability of the dam to retain the saline water. In such conditions, the rate of saline water spillage and inland extension may considerably be reduced. Conversely, the presence of an underlying low K layer led to a faster increase of saltwater volume on the seaward side of the wall, therefore considerably facilitating the spillage. The results showed that a complete removal of the residual saline water eventually occurred in all the investigated scenarios, with a rate of removal strongly affected by the hydraulic conductivity of the lower part of the aquifer. The data showed that the addition of the underlying low K layer in case HL caused the complete flushing to be almost twice longer than in the homogeneous scenario.

Keywords: heterogeneous coastal aquifers, laboratory experiments, physical barriers, seawater intrusion control

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2422 CFD Simulation of the Pressure Distribution in the Upper Airway of an Obstructive Sleep Apnea Patient

Authors: Christina Hagen, Pragathi Kamale Gurmurthy, Thorsten M. Buzug

Abstract:

CFD simulations are performed in the upper airway of a patient suffering from obstructive sleep apnea (OSA) that is a sleep related breathing disorder characterized by repetitive partial or complete closures of the upper airways. The simulations are aimed at getting a better understanding of the pathophysiological flow patterns in an OSA patient. The simulation is compared to medical data of a sleep endoscopic examination under sedation. A digital model consisting of surface triangles of the upper airway is extracted from the MR images by a region growing segmentation process and is followed by a careful manual refinement. The computational domain includes the nasal cavity with the nostrils as the inlet areas and the pharyngeal volume with an outlet underneath the larynx. At the nostrils a flat inflow velocity profile is prescribed by choosing the velocity such that a volume flow rate of 150 ml/s is reached. Behind the larynx at the outlet a pressure of -10 Pa is prescribed. The stationary incompressible Navier-Stokes equations are numerically solved using finite elements. A grid convergence study has been performed. The results show an amplification of the maximal velocity of about 2.5 times the inlet velocity at a constriction of the pharyngeal volume in the area of the tongue. It is the same region that also shows the highest pressure drop from about 5 Pa. This is in agreement with the sleep endoscopic examinations of the same patient under sedation showing complete contractions in the area of the tongue. CFD simulations can become a useful tool in the diagnosis and therapy of obstructive sleep apnea by giving insight into the patient’s individual fluid dynamical situation in the upper airways giving a better understanding of the disease where experimental measurements are not feasible. Within this study, it could been shown on one hand that constriction areas within the upper airway lead to a significant pressure drop and on the other hand a good agreement of the area of pressure drop and the area of contraction could be shown.

Keywords: biomedical engineering, obstructive sleep apnea, pharynx, upper airways

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2421 Isolation and Culture of Keratinocytes and Fibroblasts to Develop Artificial Skin Equivalent in Cats

Authors: Lavrentiadou S. N., Angelou V., Chatzimisios K., Papazoglou L.

Abstract:

The aim of this study was the isolation and culture of keratinocytes and fibroblasts from feline skin to ultimately create an artificial engineered skin (including dermis and epidermis) useful for the effective treatment of large cutaneous deficits in cats. Epidermal keratinocytes and dermal fibroblasts were freshly isolated from skin biopsies using an 8 mm biopsy punch obtained from 8 healthy cats that had undergone ovariohysterectomy. The owner’s consent was obtained. All cats had a complete blood count and a serum biochemical analysis and were screened for feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV) preoperatively. The samples were cut into small pieces and incubated with collagenase (2 mg/ml) for 5-6 hours. Following digestion, cutaneous cells were filtered through a 100 μm cell strainer, washed with DMEM, and grown in DMEM supplemented with 10% FBS. The undigested epidermis was washed with DMEM and incubated with 0.05% Trypsin/0.02% EDTA (TE) solution. Keratinocytes recovered in the TE solution were filtered through a 100 μm and a 40 μm cell strainer and, following washing, were grown on a collagen type I matrix in DMEM: F12 (3:1) medium supplemented with 10% FΒS, 1 μm hydrocortisone, 1 μm isoproterenol and 0.1 μm insulin. Both fibroblasts and keratinocytes were grown in a humidified atmosphere with 5% CO2 at 37oC. The medium was changed twice a week and cells were cultured up to passage 4. Cells were grown to 70-85% confluency, at which point they were trypsinized and subcultured in a 1:4 dilution. The majority of the cells in each passage were transferred to a freezing medium and stored at -80oC. Fibroblasts were frozen in DMEM supplemented with 30% FBS and 10% DMSO, whereas keratinocytes were frozen in a complete keratinocyte growth medium supplemented with 10% DMSO. Both cell types were thawed and successfully grown as described above. Therefore, we can create a bank of fibroblasts and keratinocytes, from which we can recover cells for further culture and use for the generation of skin equivalent in vitro. In conclusion, cutaneous cell isolation and cell culture and expansion were successfully developed. To the authors’ best knowledge, this is the first study reporting isolation and culture of keratinocytes and fibroblasts from feline skin. However, these are preliminary results and thus, the development of autologous-engineered feline skin is still in process.

Keywords: cat, fibroblasts, keratinocytes, skin equivalent, wound

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2420 Perceptions of College Students on Whether an Intelligent Tutoring System Is a Tutor

Authors: Michael Smalenberger

Abstract:

Intelligent tutoring systems (ITS) are computer-based platforms which can incorporate artificial intelligence to provide step-by-step guidance as students practice problem-solving skills. ITS can replicate the benefits of one-on-one tutoring, foster transactivity in collaborative environments, and lead to substantial learning gains when used to supplement the instruction of a teacher or when used as the sole method of instruction. Developments improving the ease of ITS creation have recently increased their proliferation, leading many K-12 schools and institutions of higher education in the United States to regularly use ITS within classrooms. We investigated how students perceive their experience using an ITS. In this study, 111 undergraduate students used an ITS in a college-level introductory statistics course and were subsequently asked for feedback on their experience. Results show that their perceptions were generally favorable of the ITS, and most would seek to use an ITS both for STEM and non-STEM courses in the future. Along with detailed transaction-level data, this feedback also provides insights on the design of user-friendly interfaces, guidance on accessibility for students with impairments, the sequencing of exercises, students’ expectation of achievement, and comparisons to other tutoring experiences. We discuss how these findings are important for the creation, implementation, and evaluation of ITS as a mode and method of teaching and learning.

Keywords: college statistics course, intelligent tutoring systems, in vivo study, student perceptions of tutoring

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2419 Antimicrobial Peptide Produced by Lactococcus garvieae with a Broad Inhibition Spectrum

Authors: Hai Chi, Ibrahim Mehmeti, Kirill Ovchinnikov, Hegle Holo, Ingolf F. Nes, Dzung B. Diep

Abstract:

By using a panel of multiple indicator strains of different bacterial species and genera, we screened a large collection of bacterial isolates (over 1800 isolates) derived from raw milk, for bacteriocin producers with broad inhibition spectra (BIS). Fourteen isolates with BIS were identified, and by 16S rDNA sequencing they were found to belong to Lactococcus garvieae (10 isolates) and Enterococcus feacalis (4 isolates). Further analysis of the ten L. garvieae isolates revealed that they were very similar, if not identical, to each other in metabolic and genetic terms: they had the same fermentation profile on different types of sugars, repetitive sequence-based PCR (rep-PCR) DNA pattern as well as they all had the same inhibition profile towards over 50 isolates of different species. The bacteriocin activity from one of the L. garvieae isolates was assessed further. The bacteriocin which was termed garvicin KS, was found to be heatstable and proteinase-labile and its inhibition spectrum contained many distantly related genera of Firmicutes, comprising most lactic acid bacteria (LAB) as well as problematic species of Bacillus, Listeria, Streptococcus and Staphylococcus and their antibiotic resistant derivatives (e.g. VRE, MRSA). Taken together, the results indicate that this is a potent bacteriocin from L. garvieae and that its very broad inhibition spectrum can be a very useful property for use in food preservation as well as in infection treatments caused by gram-positive pathogens and their antibiotic-derivatives.

Keywords: bacteriocin, lactic acid bacteria, Lactococcus garvieae, antibiotics resistance

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