Search results for: gene concordance factor confidence
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 7621

Search results for: gene concordance factor confidence

6301 Functional Switching of Serratia marcescens Transcriptional Regulator from Activator to Inhibitor of Quorum Sensing by Exogenous Addition

Authors: Norihiro Kato, Yuriko Takayama

Abstract:

Some gram-negative bacteria enable the simultaneous activation of gene expression involved in N-acylhomoserine lactone (AHL) dependent cell-to-cell communication system. Such regulatory system for the bacterial group behavior is termed as quorum sensing (QS) because a diffusible AHL signal can accumulate around the cell during the increase of the cell density and trigger activation of the sequential QS process. By blocking the QS, the expression of diverse genes related to infection, antibiotic production, and biofilm formation is inhibited. Conditioning of QS by regulation of the DNA-receptor-AHL interaction is a potential target for enhancing host defenses against pathogenicity. We focused on engineered application of transcriptional regulator SpnR produced in opportunistic human pathogen Serratia marcescens. The SpnR can interact with AHL signals at an N-terminal domain and also with a promoter region of a QS target gene at a C-terminal domain. As the initial process of the QS activation, the SpnR forms a complex with the AHL to enhance the expression of pig cluster; the SpnR normally acts as an activator for the expression of the QS-dependent gene. In this research, we attempt to artificially control QS by changing the role of SpnR. The QS-dependent prodigiosin production is expected to inhibit by externally added SpnR in the culture broth of AS-1 strain because the AHL concentration was kept below the threshold by AHL-SpnR complex formation. Maltose-binding protein (MBP)-tagged SpnR (MBP-SpnR) was overexpressed in Escherichia coli and purified using an affinity chromatography equipped with an amylose resin column. The specific interaction between AHL and MBP-SpnR was demonstrated by quartz crystal microbalance (QCM) sensor. AHL with amino end-group was coupled with COOH-terminated self-assembled monolayer prepared on a gold electrode of 27-MHz quartz crystal sensor using water-soluble carbodiimide. After the injection of MBP-SpnR into a cup-type sensor cell filled with the buffer solution, time course of resonant frequency change (ΔFs) was determined. A decrease of ΔFs clearly showed the uptake of MBP-SpnR onto the AHL-immobilized electrode. Furthermore, no binding affinity was observed after the heat-inactivation of MBP-SpnR at 80ºC. These results suggest that MBP-SpnR possesses a specific affinity for AHL. MBP-SpnR was added to the culture medium as an AHL trap to study inhibitory effects on intracellularly accumulated prodigiosin. With approximately 2 µM MBP-SpnR, the amount of prodigiosin induced was half that of the control without any additives. In conclusion, the function of SpnR could be switched by adding it to the cell culture. Exogenously added MBP-SpnR possesses high affinity for AHL derived from cells and acts as an inhibitor of AHL-mediated QS.

Keywords: intracellular signaling, microbial biotechnology, quorum sensing, transcriptional regulator

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6300 Applying Multivariate and Univariate Analysis of Variance on Socioeconomic, Health, and Security Variables in Jordan

Authors: Faisal G. Khamis, Ghaleb A. El-Refae

Abstract:

Many researchers have studied socioeconomic, health, and security variables in the developed countries; however, very few studies used multivariate analysis in developing countries. The current study contributes to the scarce literature about the determinants of the variance in socioeconomic, health, and security factors. Questions raised were whether the independent variables (IVs) of governorate and year impact the socioeconomic, health, and security dependent variables (DVs) in Jordan, whether the marginal mean of each DV in each governorate and in each year is significant, which governorates are similar in difference means of each DV, and whether these DVs vary. The main objectives were to determine the source of variances in DVs, collectively and separately, testing which governorates are similar and which diverge for each DV. The research design was time series and cross-sectional analysis. The main hypotheses are that IVs affect DVs collectively and separately. Multivariate and univariate analyses of variance were carried out to test these hypotheses. The population of 12 governorates in Jordan and the available data of 15 years (2000–2015) accrued from several Jordanian statistical yearbooks. We investigated the effect of two factors of governorate and year on the four DVs of divorce rate, mortality rate, unemployment percentage, and crime rate. All DVs were transformed to multivariate normal distribution. We calculated descriptive statistics for each DV. Based on the multivariate analysis of variance, we found a significant effect in IVs on DVs with p < .001. Based on the univariate analysis, we found a significant effect of IVs on each DV with p < .001, except the effect of the year factor on unemployment was not significant with p = .642. The grand and marginal means of each DV in each governorate and each year were significant based on a 95% confidence interval. Most governorates are not similar in DVs with p < .001. We concluded that the two factors produce significant effects on DVs, collectively and separately. Based on these findings, the government can distribute its financial and physical resources to governorates more efficiently. By identifying the sources of variance that contribute to the variation in DVs, insights can help inform focused variation prevention efforts.

Keywords: ANOVA, crime, divorce, governorate, hypothesis test, Jordan, MANOVA, means, mortality, unemployment, year

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6299 Oxygen Absorption Enhancement during Sulfite Forced Oxidation in the Presence of Nano-Particles

Authors: Zhao Bo

Abstract:

The TiO2-Na2SO3 and SiO2-Na2SO3 nano-fluids were prepared using ultrasonic dispertion method without any surfactant addition to study the influence of nano-fluids on the mass transfer during forced sulfite oxidation in a thermostatic stirred tank, and the kinetic viscosity of nano-fluids was measured. The influence of temperature (30 ℃ ~ 50 ℃), solid loading of fine particle (0 Kg/m³~1.0 Kg/m³), stirring speed (50 r/min ~ 400 r/min), and particle size (10 nm~100 nm) on the average oxygen absorption rate were investigated in detail. Both TiO2 nano-particles and SiO2 nano-particles could remarkably improve the gas-liquid mass transfer. Oxygen absorption enhancement factor increases with the increase of solid loading of nano-particles to a critical value and then decreases with further increase of solid loading under 30℃. Oxygen absorption rate together with absorption enhancement factor increases with stirring speed. However, oxygen absorption enhancement factor decreases with the increase of temperature due to aggregation of nano-particles. Further inherent relationship between particle size, loading, surface area, viscosity, stirring speed, temperature, adsorption, desorption, and mass transfer was discussed in depth by analyzing the interaction mechanism.

Keywords: fine particles, nano-fluid, mass transfer enhancement, solid loading

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6298 FEM for Stress Reduction by Optimal Auxiliary Holes in a Uniaxially Loaded Plate

Authors: Basavaraj R. Endigeri, Shriharsh Desphande

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Optimization and reduction of stress concentration around holes in a uniaxially loaded plate is one of the important design criteria in many of the engineering applications. These stress risers will lead to failure of the component at the region of high stress concentration which has to be avoided by means of providing auxiliary holes on either side of the parent hole. By literature survey it is known that till date, there is no analytical solution documented to reduce the stress concentration by providing auxiliary holes expect for fever geometries. In the present work, plate with a hole subjected to uniaxial load is analyzed with the numerical method to determine the optimum sizes and locations for the auxillary holes for different center hole diameter to plate width ratios. The introduction of auxiliary holes at a optimum location and radii with its effect on stress concentration is also represented graphically. The finite element analysis package ANSYS 8.0 is used to carry out analysis and optimization is performed to determine the location and radii for optimum values of auxiliary holes to reduce stress concentration. All the results for different diameter to plate width ratio are presented graphically. It is found from the work that introduction of auxiliary holes on either side of central circular hole will reduce stress concentration factor by a factor of 19 to 21 percentage.

Keywords: finite element method, optimization, stress concentration factor, auxiliary holes

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6297 Biochemical Characterization of CTX-M-15 from Enterobacter cloacae and Designing a Novel Non-β-Lactam-β-Lactamase Inhibitor

Authors: Mohammad Faheem, M. Tabish Rehman, Mohd Danishuddin, Asad U. Khan

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The worldwide dissemination of CTX-M type β-lactamases is a threat to human health. Previously, we have reported the spread of blaCTX-M-15 gene in different clinical strains of Enterobacteriaceae from the hospital settings of Aligarh in north India. In view of the varying resistance pattern against cephalosporins and other β-lactam antibiotics, we intended to understand the correlation between MICs and catalytic activity of CTX-M-15. In this study, steady-state kinetic parameters and MICs were determined on E. coli DH5α transformed with blaCTX-M-15 gene that was cloned from Enterobacter cloacae (EC-15) strain of clinical background. The effect of conventional β-lactamase inhibitors (clavulanic acid, sulbactam and tazobactam) on CTX-M-15 was also studied. We have found that tazobactam is the best among these inhibitors against CTX-M-15. The inhibition characteristic of tazobactam is defined by its very low IC50 value (6 nM), high affinity (Ki = 0.017 µM) and better acylation efficiency (k+2/K9 = 0.44 µM-1s-1). It forms an acyl-enzyme covalent complex, which is quite stable (k+3 = 0.0057 s-1). Since increasing resistance has been reported against conventional b-lactam antibiotic-inhibitor combinations, we aspire to design a non-b-lactam core containing b-lactamase inhibitor. For this, we screened ZINC database and performed molecular docking to identify a potential non-β-lactam based inhibitor (ZINC03787097). The MICs of cephalosporin antibiotics in combination with this inhibitor gave promising results. Steady-state kinetics and molecular docking studies showed that ZINC03787097 is a reversible inhibitor which binds non-covalently to the active site of the enzyme through hydrogen bonds and hydrophobic interactions. Though, it’s IC50 (180 nM) is much higher than tazobactam, it has good affinity for CTX-M-15 (Ki = 0.388 µM). This study concludes that ZINC03787097 compound can be used as seed molecule to design more efficient non-b-lactam containing b-lactamase inhibitor that could evade pre-existing bacterial resistance mechanisms.

Keywords: ESBL, non-b-lactam-b-lactamase inhibitor, bioinformatics, biomedicine

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6296 The Employees' Classification Method in the Space of Their Job Satisfaction, Loyalty and Involvement

Authors: Svetlana Ignatjeva, Jelena Slesareva

Abstract:

The aim of the study is development and adaptation of the method to analyze and quantify the indicators characterizing the relationship between a company and its employees. Diagnostics of such indicators is one of the most complex and actual issues in psychology of labour. The offered method is based on the questionnaire; its indicators reflect cognitive, affective and connotative components of socio-psychological attitude of employees to be as efficient as possible in their professional activities. This approach allows measure not only the selected factors but also such parameters as cognitive and behavioural dissonances. Adaptation of the questionnaire includes factor structure analysis and suitability analysis of phenomena indicators measured in terms of internal consistency of individual factors. Structural validity of the questionnaire was tested by exploratory factor analysis. Extraction Method: Principal Component Analysis. Rotation Method: Varimax with Kaiser Normalization. Factor analysis allows reduce dimension of the phenomena moving from the indicators to aggregative indexes and latent variables. Aggregative indexes are obtained as the sum of relevant indicators followed by standardization. The coefficient Cronbach's Alpha was used to assess the reliability-consistency of the questionnaire items. The two-step cluster analysis in the space of allocated factors allows classify employees according to their attitude to work in the company. The results of psychometric testing indicate possibility of using the developed technique for the analysis of employees’ attitude towards their work in companies and development of recommendations on their optimization.

Keywords: involved in the organization, loyalty, organizations, method

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6295 Teachers’ Language Insecurity in English as a Second Language Instruction: Developing Effective In-Service Training

Authors: Mamiko Orii

Abstract:

This study reports on primary school second language teachers’ sources of language insecurity. Furthermore, it aims to develop an in-service training course to reduce anxiety and build sufficient English communication skills. Language/Linguistic insecurity refers to a lack of confidence experienced by language speakers. In particular, second language/non-native learners often experience insecurity, influencing their learning efficacy. While language learner insecurity has been well-documented, research on the insecurity of language teaching professionals is limited. Teachers’ language insecurity or anxiety in target language use may adversely affect language instruction. For example, they may avoid classroom activities requiring intensive language use. Therefore, understanding teachers’ language insecurity and providing continuing education to help teachers to improve their proficiency is vital to improve teaching quality. This study investigated Japanese primary school teachers’ language insecurity. In Japan, teachers are responsible for teaching most subjects, including English, which was recently added as compulsory. Most teachers have never been professionally trained in second language instruction during college teacher certificate preparation, leading to low confidence in English teaching. Primary source of language insecurity is a lack of confidence regarding English communication skills. Their actual use of English in classrooms remains unclear. Teachers’ classroom speech remains a neglected area requiring improvement. A more refined programme for second language teachers could be constructed if we can identify areas of need. Two questionnaires were administered to primary school teachers in Tokyo: (1) Questionnaire A: 396 teachers answered questions (using a 5-point scale) concerning classroom teaching anxiety and general English use and needs for in-service training (Summer 2021); (2) Questionnaire B: 20 teachers answered detailed questions concerning their English use (Autumn 2022). Questionnaire A’s responses showed that over 80% of teachers have significant language insecurity and anxiety, mainly when speaking English in class or teaching independently. Most teachers relied on a team-teaching partner (e.g., ALT) and avoided speaking English. Over 70% of the teachers said they would like to participate in training courses in classroom English. Questionnaire B’s results showed that teachers could use simple classroom English, such as greetings and basic instructions (e.g., stand up, repeat after me), and initiate conversation (e.g., asking questions). In contrast, teachers reported that conversations were mainly carried on in a simple question-answer style. They had difficulty continuing conversations. Responding to learners’ ‘on-the-spot’ utterances was particularly difficult. Instruction in turn-taking patterns suitable in the classroom communication context is needed. Most teachers received grammar-based instruction during their entire English education. They were predominantly exposed to displayed questions and form-focused corrective feedback. Therefore, strategies such as encouraging teachers to ask genuine questions (i.e., referential questions) and responding to students with content feedback are crucial. When learners’ utterances are incorrect or unsatisfactory, teachers should rephrase or extend (recast) them instead of offering explicit corrections. These strategies support a continuous conversational flow. These results offer benefits beyond Japan’s English as a second Language context. They will be valuable in any context where primary school teachers are underprepared but must provide English-language instruction.

Keywords: english as a second/non-native language, in-service training, primary school, teachers’ language insecurity

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6294 Characteristic Study on Conventional and Soliton Based Transmission System

Authors: Bhupeshwaran Mani, S. Radha, A. Jawahar, A. Sivasubramanian

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Here, we study the characteristic feature of conventional (ON-OFF keying) and soliton based transmission system. We consider 20 Gbps transmission system implemented with Conventional Single Mode Fiber (C-SMF) to examine the role of Gaussian pulse which is the characteristic of conventional propagation and hyperbolic-secant pulse which is the characteristic of soliton propagation in it. We note the influence of these pulses with respect to different dispersion lengths and soliton period in conventional and soliton system, respectively, and evaluate the system performance in terms of quality factor. From the analysis, we could prove that the soliton pulse has more consistent performance even for long distance without dispersion compensation than the conventional system as it is robust to dispersion. For the length of transmission of 200 Km, soliton system yielded Q of 33.958 while the conventional system totally exhausted with Q=0.

Keywords: dispersion length, retrun-to-zero (rz), soliton, soliton period, q-factor

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6293 Biomedical Application of Green Biosynthesis Magnetic Iron Oxide (Fe3O4) Nanoparticles Using Seaweed (Sargassum muticum) Aqueous Extract

Authors: Farideh Namvar, Rosfarizan Mohamed

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In the field of nanotechnology, the use of various biological units instead of toxic chemicals for the reduction and stabilization of nanoparticles, has received extensive attention. This use of biological entities to create nanoparticles has designated as “Green” synthesis and it is considered to be far more beneficial due to being economical, eco-friendly and applicable for large-scale synthesis as it operates on low pressure, less input of energy and low temperatures. The lack of toxic byproducts and consequent decrease in degradation of the product renders this technique more preferable over physical and classical chemical methods. The variety of biomass having reduction properties to produce nanoparticles makes them an ideal candidate for fabrication. Metal oxide nanoparticles have been said to represent a "fundamental cornerstone of nanoscience and nanotechnology" due to their variety of properties and potential applications. However, this also provides evidence of the fact that metal oxides include many diverse types of nanoparticles with large differences in chemical composition and behaviour. In this study, iron oxide nanoparticles (Fe3O4-NPs) were synthesized using a rapid, single step and completely green biosynthetic method by reduction of ferric chloride solution with brown seaweed (Sargassum muticum) water extract containing polysaccharides as a main factor which acts as reducing agent and efficient stabilizer. Antimicrobial activity against six microorganisms was tested using well diffusion method. The resulting S-IONPs are crystalline in nature, with a cubic shape. The average particle diameter, as determined by TEM, was found to be 18.01 nm. The S-IONPs were efficiently inhibited the growth of Listeria monocytogenes, Escherichia coli and Candida species. Our favorable results suggest that S-IONPs could be a promising candidate for development of future antimicrobial therapies. The nature of biosynthesis and the therapeutic potential by S-IONPs could pave the way for further research on design of green synthesis therapeutic agents, particularly nanomedicine, to deal with treatment of infections. Further studies are needed to fully characterize the toxicity and the mechanisms involved with the antimicrobial activity of these particles. Antioxidant activity of S-IONPs synthesized by green method was measured by ABTS (2, 2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (IC50= 1000µg) radical scavenging activity. Also, with the increasing concentration of S-IONPs, catalase gene expression compared to control gene GAPDH increased. For anti-angiogenesis study the Ross fertilized eggs were divided into four groups; the control and three experimental groups. The gelatin sponges containing albumin were placed on the chorioalantoic membrane and soaked with different concentrations of S-IONPs. All the cases were photographed using a photo stereomicroscope. The number and the lengths of the vessels were measured using Image J software. The crown rump (CR) and weight of the embryo were also recorded. According to the data analysis, the number and length of the blood vessels, as well as the CR and weight of the embryos reduced significantly compared to the control (p < 0.05), dose dependently. The total hemoglobin was quantified as an indicator of the blood vessel formation, and in the treated samples decreased, which showed its inhibitory effect on angiogenesis.

Keywords: anti-angiogenesis, antimicrobial, antioxidant, biosynthesis, iron oxide (fe3o4) nanoparticles, sargassum muticum, seaweed

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6292 Fabrication of Electrospun Green Fluorescent Protein Nano-Fibers for Biomedical Applications

Authors: Yakup Ulusu, Faruk Ozel, Numan Eczacioglu, Abdurrahman Ozen, Sabriye Acikgoz

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GFP discovered in the mid-1970s, has been used as a marker after replicated genetic study by scientists. In biotechnology, cell, molecular biology, the GFP gene is frequently used as a reporter of expression. In modified forms, it has been used to make biosensors. Many animals have been created that express GFP as an evidence that a gene can be expressed throughout a given organism. Proteins labeled with GFP identified locations are determined. And so, cell connections can be monitored, gene expression can be reported, protein-protein interactions can be observed and signals that create events can be detected. Additionally, monitoring GFP is noninvasive; it can be detected by under UV-light because of simply generating fluorescence. Moreover, GFP is a relatively small and inert molecule, that does not seem to treat any biological processes of interest. The synthesis of GFP has some steps like, to construct the plasmid system, transformation in E. coli, production and purification of protein. GFP carrying plasmid vector pBAD–GFPuv was digested using two different restriction endonuclease enzymes (NheI and Eco RI) and DNA fragment of GFP was gel purified before cloning. The GFP-encoding DNA fragment was ligated into pET28a plasmid using NheI and Eco RI restriction sites. The final plasmid was named pETGFP and DNA sequencing of this plasmid indicated that the hexa histidine-tagged GFP was correctly inserted. Histidine-tagged GFP was expressed in an Escherichia coli BL21 DE3 (pLysE) strain. The strain was transformed with pETGFP plasmid and grown on LuiraBertoni (LB) plates with kanamycin and chloramphenicol selection. E. coli cells were grown up to an optical density (OD 600) of 0.8 and induced by the addition of a final concentration of 1mM isopropyl-thiogalactopyranoside (IPTG) and then grown for additional 4 h. The amino-terminal hexa-histidine-tag facilitated purification of the GFP by using a His Bind affinity chromatography resin (Novagen). Purity of GFP protein was analyzed by a 12 % sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The concentration of protein was determined by UV absorption at 280 nm (Varian Cary 50 Scan UV/VIS spectrophotometer). Synthesis of GFP-Polymer composite nanofibers was produced by using GFP solution (10mg/mL) and polymer precursor Polyvinylpyrrolidone, (PVP, Mw=1300000) as starting materials and template, respectively. For the fabrication of nanofibers with the different fiber diameter; a sol–gel solution comprising of 0.40, 0.60 and 0.80 g PVP (depending upon the desired fiber diameter) and 100 mg GFP in 10 mL water: ethanol (3:2) mixtures were prepared and then the solution was covered on collecting plate via electro spinning at 10 kV with a feed-rate of 0.25 mL h-1 using Spellman electro spinning system. Results show that GFP-based nano-fiber can be used plenty of biomedical applications such as bio-imaging, bio-mechanic, bio-material and tissue engineering.

Keywords: biomaterial, GFP, nano-fibers, protein expression

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6291 First Report of Rahnella Victoriana Associated with Walnut Decline

Authors: Mohammadreza Hajialigol, Nargues Falahi Charkhabi, Fatemeh Shahryari, Saadat Sarikhani

Abstract:

BACKGROUND AND OBJECTIVES Iran is the third producer of Persian walnut worldwide. However, its walnut trees have been under threat from decline during last decade. Walnut canker caused by B. nigrifluens and B. rubrifaciens was recorded in multiple regions of Iran. Furthermore, Brenneria rosae subsp. rosae and Gibbsiella quercinecans were recently recognized as responsible for walnut decline in northwestern Iran. This study aimed to identify the causal agent of walnut decline in Kermanshah and Isfahan. MATERIAL AND METHODS Symptomatic samples were collected from affected walnut trees of Kermanshah and Isfahan provinces. The pathogenicity of strains was proved on immature walnut fruits cv. ‘Hartley’ and young green twigs of two-year-old walnut seedling cv. ‘Chandler’. Pathogenic strains were subjected to conventional phenotypic tests. 16S rRNA, gyrB, and infB genes were partially amplified and sequenced. RESULTS Irregular longitudinal cankers and dark lesions were observed in the outer and inner bark, respectively. Twenty-four strains were isolated on EMB-agar media. Fourteen strains were able to cause necrosis and a dark-colored region in the mesocarp and on young green twigs around the inoculation site 14 and 30 days post-inoculation, respectively. Strains were able to hydrolyze Tween 20, Tween 80, gelatin and esculin, however, did not produce indole or urease. Pairwise comparison, the 16S rRNA gene nucleotide sequences of strain I2 were 100% identical with those of Rahnella victoriana FRB 225T. Moreover, a phylogenetic tree reconstructed based on the concatenated sequences of two housekeeping gene fragments, gyrB (601 bp) and infB (615 bp), revealed that the strains I2, I5, and KE6 were clustered with R. victoriana FRB 225T. CONCLUSION To the best of our knowledge, this is the first report of R. victoriana in association with walnut decline. This result is necessary to find resistant genotypes.

Keywords: emerging pathogens, Iran, juglans regia, MLSA

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6290 Biodegradation of 2,4-Dichlorophenol by Pseudomonas chlororaphis Strain Isolated from Activated Sludge Sample from a Wastewater Treatment Plant in Durban, South Africa

Authors: Boitumelo Setlhare, Mduduzi P. Mokoena, Ademola O. Olaniran

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Agricultural and industrial activities have led to increasing production of xenobiotics such as 2,4-dichlorophenol (2,4-DCP), a derivative of 2,4-dichlorophenoxyacetic acid (2,4-D), which is a widely used herbicide. Bioremediation offers an efficient, cost-effective and environmentally friendly method for degradation of the compound through the activities of the various microbial enzymes involved in the catabolic pathway. The aim of this study was to isolate and characterize bacterial isolate indigenous to contaminated sites in Durban, South Africa for 2,4-DCP degradation. One bacterium capable of utilizing 2,4-DCP as sole carbon source was isolated using culture enrichment technique and identified as Pseudomonas chlororaphis strain UFB2 via PCR amplification and analysis of 16S rRNA gene sequence. This isolate was able to degrade up to 75.11% of 2,4-DCP in batch cultures within 10 days, with the degradation rate constant of 0.14 mg/l/d. Phylogenetic analysis revealed the relatedness of this bacterial isolate to other Pseudomonas sp. previously characterized for chlorophenol degradation. PCR amplification of the catabolic genes involved in 2,4-DCP degradation revealed the presence of the correct amplicons for phenol hydroxylase (600 bp), catechol 1,2-dioxygenase (214 bp), muconate isomerase (851 bp), cis-dienelactone hydrolase (577 bp), and trans-dienelactone hydrolase (491 bp) genes. Enzyme assays revealed activity as high as 21840 mU/mg, 15630 mU/mg, 2340 mU/mg and 1490 mU/mg obtained for phenol hydroxylase, catechol 1,2-dioxygenase, cis-dienelactone hydroxylase and trans-dienelactone hydroxylase, respectively. The absence of catechol 2,3-dioxygenase gene and the corresponding enzyme in this isolate suggests that the organism followed ortho-pathway for 2,4-DCP degradation. Furthermore, the absence of malaycetate reductase genes showed that the bacterium may not be able to completely mineralize 2,4-DCP. Further studies are required to optimize 2,4-DCP degradation by this isolate as well as to elucidate the mechanism of 2,4-DCP degradation.

Keywords: biodegradation, catechol 1, 2-dioxygenase, 2, 4-dichlorophenol, phenol hydroxylase, Pseudomonas chlororaphis

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6289 Effect of the Diverse Standardized Patient Simulation Cultural Competence Education Strategy on Nursing Students' Transcultural Self-Efficacy Perceptions

Authors: Eda Ozkara San

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Nurse educators have been charged by several nursing organizations and accrediting bodies to provide innovative and evidence-based educational experiences, both didactic and clinical, to help students to develop the knowledge, skills, and attitudes needed to provide culturally competent nursing care to patients. Clinical simulation, which offers the opportunity for students to practice nursing skills in a risk-free, controlled environment and helps develop self-efficacy (confidence) within the nursing role. As one simulation method, the standardized patients (SPs) simulation helps educators to teach nursing students variety of skills in nursing, medicine, and other health professions. It can be a helpful tool for nurse educators to enhance cultural competence of nursing students. An alarming gap exists within the literature concerning the effectiveness of SP strategy to enhance cultural competence development of diverse student groups, who must work with patients from various backgrounds. This grant-supported, longitudinal, one-group, pretest and post-test educational intervention study aimed to examine the effect of the Diverse Standardized Patient Simulation (DSPS) cultural competence education strategy on students’ (n = 53) transcultural self-efficacy (TSE). The researcher-developed multidimensional DSPS strategy involved careful integration of transcultural nursing skills guided by the Cultural Competence and Confidence (CCC) model. As a carefully orchestrated teaching and learning strategy by specifically utilizing the SP pedagogy, the DSPS also followed international guidelines and standards for the design, implementation, evaluation, and SP training; and had content validity review. The DSPS strategy involved two simulation scenarios targeting underrepresented patient populations (Muslim immigrant woman with limited English proficiency and Irish-Italian American gay man with his partner (Puerto Rican) to be utilized in a second-semester, nine-credit, 15-week medical-surgical nursing course at an urban public US university. Five doctorally prepared content experts reviewed the DSPS strategy for content validity. The item-level content validity index (I-CVI) score was calculated between .80-1.0 on the evaluation forms. Jeffreys’ Transcultural Self-Efficacy Tool (TSET) was administered as a pretest and post-test to assess students’ changes in cognitive, practical, and affective dimensions of TSE. Results gained from this study support that the DSPS cultural competence education strategy assisted students to develop cultural competence and caused statistically significant changes (increase) in students’ TSE perceptions. Results also supported that all students, regardless of their background, benefit (and require) well designed cultural competence education strategies. The multidimensional DSPS strategy is found to be an effective way to foster nursing students’ cultural competence development. Step-by-step description of the DSPS provides an easy adaptation of this strategy with different student populations and settings.

Keywords: cultural competence development, the cultural competence and confidence model, CCC model, educational intervention, transcultural self-efficacy, TSE, transcultural self-efficacy tool, TSET

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6288 The Effect of the Contributory Pension Scheme on Employees’ Performance

Authors: Oladipo Jimoh Ayanda, Fashagba Mathew Olasehinde

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Pension is a post retirement benefit paid to employees after retirement to cushion the effects of severance from monthly emoluments. It serves the dual purpose of providing financial succour to retired employees as well as motivating employees currently in service to greater performance on duty. However, the scheme, as operated in Nigeria, is prone to some pitfalls such as delayed and irregular payments, inadequate budgetary provisions, employee sufferings and deaths arising from the rigors of verification exercises, among others. This necessitated the replacement of the old scheme with the contributory pension scheme through an enabling law in 2004. The implementation of the new scheme has its own challenges especially in connection with administration. These challenges pose a fundamental problem of establishing a nexus between pension benefits and work performance which represent the focus of the study. The study objectives were to: determine the effect of contributory pension scheme on employees’ performance. The study population consisted of National Universities Commission recognized public and private universities in the South West Nigeria. Multi-stage sampling method involving stratified sampling and systematic sampling was used in selecting 359 respondents while data were collected through questionnaire administration. The procedure for analyzing the data included descriptive statistic, normal distribution test and cross-tabulation (gamma coefficient). The findings of the study showed that the existence of the scheme positively enhances employees’ performance as indicated by normal distribution test with Z-score (10.169) which is greater than the table value (1.96) at 0.05 level. The study concluded that the scope for enhancing employee current job performance can be quite elastic if future retirement benefits are guaranteed through proper and efficient administration and management of the contributory pension scheme. The study recommended that certain factors such as employers’ commitment which account for different levels of confidence between public and private universities should be looked into in order to improve confidence across board while the provisions of the scheme as they affect the PFAs should be properly monitored to ensure compliance.

Keywords: pension, retirement, performance, employees, benefit

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6287 Persistent Ribosomal In-Frame Mis-Translation of Stop Codons as Amino Acids in Multiple Open Reading Frames of a Human Long Non-Coding RNA

Authors: Leonard Lipovich, Pattaraporn Thepsuwan, Anton-Scott Goustin, Juan Cai, Donghong Ju, James B. Brown

Abstract:

Two-thirds of human genes do not encode any known proteins. Aside from long non-coding RNA (lncRNA) genes with recently-discovered functions, the ~40,000 non-protein-coding human genes remain poorly understood, and a role for their transcripts as de-facto unconventional messenger RNAs has not been formally excluded. Ribosome profiling (Riboseq) predicts translational potential, but without independent evidence of proteins from lncRNA open reading frames (ORFs), ribosome binding of lncRNAs does not prove translation. Previously, we mass-spectrometrically documented translation of specific lncRNAs in human K562 and GM12878 cells. We now examined lncRNA translation in human MCF7 cells, integrating strand-specific Illumina RNAseq, Riboseq, and deep mass spectrometry in biological quadruplicates performed at two core facilities (BGI, China; City of Hope, USA). We excluded known-protein matches. UCSC Genome Browser-assisted manual annotation of imperfect (tryptic-digest-peptides)-to-(lncRNA-three-frame-translations) alignments revealed three peptides hypothetically explicable by 'stop-to-nonstop' in-frame replacement of stop codons by amino acids in two ORFs of the lncRNA MMP24-AS1. To search for this phenomenon genomewide, we designed and implemented a novel pipeline, matching tryptic-digest spectra to wildcard-instead-of-stop versions of repeat-masked, six-frame, whole-genome translations. Along with singleton putative stop-to-nonstop events affecting four other lncRNAs, we identified 24 additional peptides with stop-to-nonstop in-frame substitutions from multiple positive-strand MMP24-AS1 ORFs. Only UAG and UGA, never UAA, stop codons were impacted. All MMP24-AS1-matching spectra met the same significance thresholds as high-confidence known-protein signatures. Targeted resequencing of MMP24-AS1 genomic DNA and cDNA from the same samples did not reveal any mutations, polymorphisms, or sequencing-detectable RNA editing. This unprecedented apparent gene-specific violation of the genetic code highlights the importance of matching peptides to whole-genome, not known-genes-only, ORFs in mass-spectrometry workflows, and suggests a new mechanism enhancing the combinatorial complexity of the proteome. Funding: NIH Director’s New Innovator Award 1DP2-CA196375 to LL.

Keywords: genetic code, lncRNA, long non-coding RNA, mass spectrometry, proteogenomics, ribo-seq, ribosome, RNAseq

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6286 Breast Cancer in Very Young (Less Than 25 Yeras) Women: An Institutional Analysis from Developing Country

Authors: Ajay Gogia, Svs Deo, Dn Sharma, Atul Batra, Ashutash Mishra

Abstract:

Background and Aims: Breast cancer in women aged less than 25 years (defined as very young breast cancer, VYBC) is rare and accounts for 0.25% of all breast cancer in the West. There is no data available on VYBC from developing countries. The aim of this study was to analyze the clinical, pathological, and prognostic factors and outcomes in VYBC. Methods: This retrospective analysis was performed on 80 patients aged 25 years or less (screened 8000 files of female BC) who were registered at All India Institute of Medical Sciences (AIIMS), New Delhi, India, over a 15-year period between 2011 and 2023. Results: The median age was 21.5 years (range 16-25). A positive family history (siblings and parents) was elicited in 30% of cases, and breast cancer gene (BRCA1/2) mutation was found in 33% of cases patients. Ten patients (12.5%) patients have pregnancy-associated breast cancer (BC detected during pregnancy or 1 year after postpartum period). The TNM stage distribution was Stage I was 0, stage II -30%, stage III –60% and Stage IV -10 %patients. Seventy percent of tumors were high grade, and 90% had pathological node-positive disease. Estrogen, Progesterone, and human epidermal growth factor receptor 2 (HER2)/neu positivity were 25%,25% and 35%, respectively. Triple-negative breast cancer constituted 40% of patients. With a median follow-up of 42 months, 3 years, relapse-free survival (nonmetastatic disease), progression-free survival (metastatic disease) and overall survival were 30%, 15% and 50%, respectively. Conclusions: Very young women constituted 1% of all breast cancer cases. Advanced disease at presentation and high-risk pathological features result in poor outcomes. One-third of VYBCs are associated with BRCA mutation, which requires genetic counseling and risk reduction surgery if required. Due to the aggressive behavior of BC in this age group, need early diagnosis and prompt treatment

Keywords: very young, breast cancer, outcome, developing country, India

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6285 Identification of the Key Enzyme of Roseoflavin Biosynthesis

Authors: V. Konjik, J. Schwartz, R. Sandhoff, M. Mack

Abstract:

The rising number of multi-resistant pathogens demands the development of new antibiotics in order to reduce the lethal risk of infections. Here, we investigate roseoflavin, a vitamin B2 analogue which is produced by Streptomyces davawensis and Streptomyces cinnabarinus. We consider roseoflavin to be a 'Trojan horse' compound. Its chemical structure is very similar to riboflavin but in fact it is a toxin. Furthermore, it is a clever strategy with regard to the delivery of an antibiotic to its site of action but also with regard to the production of this chemical: The producer cell has only to convert a vitamin (which is already present in the cytoplasm) into a vitamin analog. Roseoflavin inhibits the activity of Flavin depending proteins, which makes up to 3.5 % of predicted proteins in organisms sequenced so far. We sequentially knocked out gene clusters and later on single genes in order to find the ones which are involved in the roseoflavin biosynthesis. Consequently, we identified the gene rosB, coding for the protein carrying out the first step of roseoflavin biosynthesis, starting form Flavin mononucleotide. Here we show, that the protein RosB has so far unknown features. It is per se an oxidoreductase, a decarboxylase and an aminotransferase, all rolled into one enzyme. A screen of cofactors revealed needs of oxygen, NAD+, thiamine and glutamic acid to carry out its function. Surprisingly, thiamine is not only needed for the decaboxylation step, but also for the oxidation of 8-demethyl-8-formyl Flavin mononucleotide. We had managed to isolate three different Flavin intermediates with different oxidation states, which gave us a mechanistic insight of RosB functionality. Our work points to a so far new function of thiamine in Streptomyces davawensis. Additionally, RosB could be extremely useful for chemical synthesis. Careful engineering of RosB may allow the site-specific replacement of methyl groups by amino groups in polyaromatic compounds of commercial interest. Finally, the complete clarification of the roseoflavin biosynthesis opens the possibility of engineering cost-effective roseoflavin producing strains.

Keywords: antibiotic, flavin analogue, roseoflavin biosynthesis, vitamin B2

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6284 Relational and Personal Variables Predicting Marital Satisfaction

Authors: Sezen Gulec, Bilge Uzun

Abstract:

Almost all of the world population marries at least once in their lifetime. Nevertheless, in reality, only half of all marriages last a lifetime. The most important factor in marriage to manage is the satisfaction that they obtain. It is reality that marital satisfaction does not only related to maintain the relationship but also related to the social and work relationships. In this respect, the purpose of the present research is to find the personal and relational factors predicted marital satisfaction. The sample including 378 (178 male and 200 females) married individuals were administered to marital life scale, multidimensional perfectionism scale, trait forgivingness scale, adjective based personality test and relationship happiness questionnaire. The findings revealed marital happiness, forgiveness and extravertedness and emotional inconsistency factors were found to be significant predictors of marital satisfaction.

Keywords: marital satisfaction, happiness, perfectionism, forgiveness, five factor personality

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6283 Atmospheric Circulation Drivers Of Nationally-Aggregated Wind Energy Production Over Greece

Authors: Kostas Philippopoulos, Chris G. Tzanis, Despina Deligiorgi

Abstract:

Climate change adaptation requires the exploitation of renewable energy sources such as wind. However, climate variability can affect the regional wind energy potential and consequently the available wind power production. The goal of the research project is to examine the impact of atmospheric circulation on wind energy production over Greece. In the context of synoptic climatology, the proposed novel methodology employs Self-Organizing Maps for grouping and classifying the atmospheric circulation and nationally-aggregated capacity factor time series for a 30-year period. The results indicate the critical effect of atmospheric circulation on the national aggregated wind energy production values and therefore address the issue of optimum distribution of wind farms for a specific region.

Keywords: wind energy, atmospheric circulation, capacity factor, self-organizing maps

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6282 Development and Validation of Sense of Humor Questionnaire in China

Authors: Yunshi Peng, Shanshan Gao, Sang Qin

Abstract:

The sense of humor is an integration of cognition, emotion and behavioral tendencies in the process of expressing humor. Previous studies evidenced the positive impact of sense of humor on promoting mental health. However, very few studies investigated this with Chinese populations. The absence of a validated questionnaire limits empirical research on sense of humor in China. This study aimed to develop a Chinese instrument to examine the sense of humor among college students in China. A pool of 72 items was developed by conducting a series of qualitative methods including open-ended questionnaire, individual interviews and literature analysis, followed by an expert rating. A total of 500 college students were recruited from 7 provinces in China to complete all 72 items. The factor structure of sense of humor was established and 25 items were eventually formed by utilizing the exploratory factor analyses (EFA). The questionnaire composed 4 subscales: humor comprehension, humor creativity, attitudes towards humor and optimism level. Confirmatory factor analyses (CFA) from a follow-up study with a different sample of 1200 colleges students showed good model fit. All subscales and the overall questionnaire display satisfying internal consistency. Correlations with criterion variables demonstrated good convergent and discriminant validity. The sense of humor questionnaire is a psychometrically-sound instrument for the population of college students in China. This is applicable for future studies to identify the structure of sense of humor and evaluate the levels of humor for individuals.

Keywords: college students, EFA and CFA, questionnaire, sense of humor

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6281 Surgical Skills in Mulanje

Authors: Nick Toossi, Joseph Hartland

Abstract:

Background: Malawi is an example of a low resource setting which faces a chronic shortage of doctors and other medical staff. This shortfall is made up for by clinical officers (COs), who are para-medicals trained for 4 years. The literature suggests to improve outcomes surgical skills training specifically should be promoted for COs in district and mission hospitals. Accordingly, the primary author was tasked with developing a basic surgical skills teaching package for COs of Mulanje Mission Hospital (MMH), Malawi, as part of a 4th year medical student External Student Selected Component field trip. MMH is a hospital based in the South of Malawi near the base of Mulanje Mountain and works in an extremely isolated environment with some of the poorest communities in the country. Traveling to Malawi the medical student author performed an educational needs assessment to develop and deliver a bespoke basic surgical skills teaching package. Methodology: An initial needs assessment identified the following domains: basic surgical skills (instrument naming & handling, knot tying, suturing principles and suturing techniques) and perineal repair. Five COs took part in a teaching package involving an interactive group simulation session, overseen by senior clinical officers and surgical trainees from the UK. Non-organic and animal models were used for simulation practice. This included the use of surgical skills boards to practice knot tying and ox tongue to simulate perineal repair. All participants spoke and read English. The impact of the session was analysed in two different ways. The first was via a pre and post Single Best Answer test and the second a questionnaire including likert’s scales and free text response questions. Results: There was a positive trend in pre and post test scores on competition of the course. There was increase in the mean confidence of learners before and after the delivery of teaching in basic surgical skills and simulated perineal repair, especially in ‘instrument naming and handling’. Whilst positively received it was discovered that learners desire more frequent surgical skills teaching sessions in order to improve and revise skills. Feedback suggests that the learners were not confident in retaining the skills without regular input. Discussion: Skills and confidence were improved as a result of the teaching provided. Learner's written feedback suggested there was an overall appetite for regular surgical skills teaching in the clinical environment and further opportunities to allow for deliberate self-practice. Surgical mentorship schemes facilitating supervised theatre time among trainees and lead surgeons along with improving access to surgical models/textbooks were some of the simple suggestions to improve surgical skills and confidence among COs. Although, this study is limited by population size it is reflective of the small, isolated and low resource environment in which this healthcare is delivered. This project does suggest that current surgical skills packages used in the UK could be adapted for employment in low resource settings, but it is consistency and sustainability that staff seek above all in their on-going education.

Keywords: clinical officers, education, Malawi, surgical skills

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6280 Electron Density Discrepancy Analysis of Energy Metabolism Coenzymes

Authors: Alan Luo, Hunter N. B. Moseley

Abstract:

Many macromolecular structure entries in the Protein Data Bank (PDB) have a range of regional (localized) quality issues, be it derived from x-ray crystallography, Nuclear Magnetic Resonance (NMR) spectroscopy, or other experimental approaches. However, most PDB entries are judged by global quality metrics like R-factor, R-free, and resolution for x-ray crystallography or backbone phi-psi distribution statistics and average restraint violations for NMR. Regional quality is often ignored when PDB entries are re-used for a variety of structurally based analyses. The binding of ligands, especially ligands involved in energy metabolism, is of particular interest in many structurally focused protein studies. Using a regional quality metric that provides chemically interpretable information from electron density maps, a significant number of outliers in regional structural quality was detected across x-ray crystallographic PDB entries for proteins bound to biochemically critical ligands. In this study, a series of analyses was performed to evaluate both specific and general potential factors that could promote these outliers. In particular, these potential factors were the minimum distance to a metal ion, the minimum distance to a crystal contact, and the isotropic atomic b-factor. To evaluate these potential factors, Fisher’s exact tests were performed, using regional quality criteria of outlier (top 1%, 2.5%, 5%, or 10%) versus non-outlier compared to a potential factor metric above versus below a certain outlier cutoff. The results revealed a consistent general effect from region-specific normalized b-factors but no specific effect from metal ion contact distances and only a very weak effect from crystal contact distance as compared to the b-factor results. These findings indicate that no single specific potential factor explains a majority of the outlier ligand-bound regions, implying that human error is likely as important as these other factors. Thus, all factors, including human error, should be considered when regions of low structural quality are detected. Also, the downstream re-use of protein structures for studying ligand-bound conformations should screen the regional quality of the binding sites. Doing so prevents misinterpretation due to the presence of structural uncertainty or flaws in regions of interest.

Keywords: biomacromolecular structure, coenzyme, electron density discrepancy analysis, x-ray crystallography

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6279 Serum Granulocyte Colony Stimulating Factor is a Potent Stimulator of Hematopoeitic Progenitor Cells Mobilization in Trauma Hemorrhagic Shock

Authors: Manoj Kumar, Sujata Mohanty, D. N. Rao, Arul Selvi, Sanjeev K. Bhoi

Abstract:

Background: Hematopoietic progenitor cells (HPC) mobilized from bone marrow to peripheral blood has been observed in severe trauma and hemorrhagic shock patients. Granulocyte-colony stimulating factor (G-CSF) is a potent stimulator that mobilized HPC from bone marrow to peripheral blood. Objective: Our aim of the study was to investigate the serum G-CSF levels and correlate with HPC and outcome. Methods: Peripheral blood sample from 50 hemorrhagic shock patients was collected on arrival for determination of G-CSF and peripheral blood HPC (PBHPC) and compared with healthy control (n=15). Determination of serum levels of G-CSF by sandwich ELISA and PBHPC by Sysmex XE-2100. Data were categorized by age, sex, Injury Severity Score (ISS), and laboratory data was prospectively collected. Data are expressed as mean±SD and median (min, max). Results: Significantly increased the serum level of G-CSF (264.8 vs. 79.1 pg/ml) and peripheral blood HPC (0.1 vs. 0.01 %) in the T/HS patients when compared with control group. Conclusions: Our studies suggest serum G-CSF elevated in T/HS patients. The elevated in G-CSF was also associated with mobilization of HPC from BM to peripheral blood HPC. Increased the levels of G-CSF in T/HS may play a significant role in the alteration of the hematopoietic compartment.

Keywords: granulocyte colony stimulating factor, G-CSF, hematopoietic progenitor cells, HPC, trauma hemorrhagic shock, T/HS, outcome

Procedia PDF Downloads 332
6278 Single Pass Design of Genetic Circuits Using Absolute Binding Free Energy Measurements and Dimensionless Analysis

Authors: Iman Farasat, Howard M. Salis

Abstract:

Engineered genetic circuits reprogram cellular behavior to act as living computers with applications in detecting cancer, creating self-controlling artificial tissues, and dynamically regulating metabolic pathways. Phenemenological models are often used to simulate and design genetic circuit behavior towards a desired behavior. While such models assume that each circuit component’s function is modular and independent, even small changes in a circuit (e.g. a new promoter, a change in transcription factor expression level, or even a new media) can have significant effects on the circuit’s function. Here, we use statistical thermodynamics to account for the several factors that control transcriptional regulation in bacteria, and experimentally demonstrate the model’s accuracy across 825 measurements in several genetic contexts and hosts. We then employ our first principles model to design, experimentally construct, and characterize a family of signal amplifying genetic circuits (genetic OpAmps) that expand the dynamic range of cell sensors. To develop these models, we needed a new approach to measuring the in vivo binding free energies of transcription factors (TFs), a key ingredient of statistical thermodynamic models of gene regulation. We developed a new high-throughput assay to measure RNA polymerase and TF binding free energies, requiring the construction and characterization of only a few constructs and data analysis (Figure 1A). We experimentally verified the assay on 6 TetR-homolog repressors and a CRISPR/dCas9 guide RNA. We found that our binding free energy measurements quantitatively explains why changing TF expression levels alters circuit function. Altogether, by combining these measurements with our biophysical model of translation (the RBS Calculator) as well as other measurements (Figure 1B), our model can account for changes in TF binding sites, TF expression levels, circuit copy number, host genome size, and host growth rate (Figure 1C). Model predictions correctly accounted for how these 8 factors control a promoter’s transcription rate (Figure 1D). Using the model, we developed a design framework for engineering multi-promoter genetic circuits that greatly reduces the number of degrees of freedom (8 factors per promoter) to a single dimensionless unit. We propose the Ptashne (Pt) number to encapsulate the 8 co-dependent factors that control transcriptional regulation into a single number. Therefore, a single number controls a promoter’s output rather than these 8 co-dependent factors, and designing a genetic circuit with N promoters requires specification of only N Pt numbers. We demonstrate how to design genetic circuits in Pt number space by constructing and characterizing 15 2-repressor OpAmp circuits that act as signal amplifiers when within an optimal Pt region. We experimentally show that OpAmp circuits using different TFs and TF expression levels will only amplify the dynamic range of input signals when their corresponding Pt numbers are within the optimal region. Thus, the use of the Pt number greatly simplifies the genetic circuit design, particularly important as circuits employ more TFs to perform increasingly complex functions.

Keywords: transcription factor, synthetic biology, genetic circuit, biophysical model, binding energy measurement

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6277 Tip60’s Novel RNA-Binding Function Modulates Alternative Splicing of Pre-mRNA Targets Implicated in Alzheimer’s Disease

Authors: Felice Elefant, Akanksha Bhatnaghar, Keegan Krick, Elizabeth Heller

Abstract:

Context: The severity of Alzheimer’s Disease (AD) progression involves an interplay of genetics, age, and environmental factors orchestrated by histone acetyltransferase (HAT) mediated neuroepigenetic mechanisms. While disruption of Tip60 HAT action in neural gene control is implicated in AD, alternative mechanisms underlying Tip60 function remain unexplored. Altered RNA splicing has recently been highlighted as a widespread hallmark in the AD transcriptome that is implicated in the disease. Research Aim: The aim of this study was to identify a novel RNA binding/splicing function for Tip60 in human hippocampus and impaired in brains from AD fly models and AD patients. Methodology/Analysis: The authors used RNA immunoprecipitation using RNA isolated from 200 pooled wild type Drosophila brains for each of the 3 biological replicates. To identify Tip60’s RNA targets, they performed genome sequencing (DNB-SequencingTM technology, BGI genomics) on 3 replicates for Input RNA and RNA IPs by Tip60. Findings: The authors' transcriptomic analysis of RNA bound to Tip60 by Tip60-RNA immunoprecipitation (RIP) revealed Tip60 RNA targets enriched for critical neuronal processes implicated in AD. Remarkably, 79% of Tip60’s RNA targets overlap with its chromatin gene targets, supporting a model by which Tip60 orchestrates bi-level transcriptional regulation at both the chromatin and RNA level, a function unprecedented for any HAT to date. Since RNA splicing occurs co-transcriptionally and splicing defects are implicated in AD, the authors investigated whether Tip60-RNA targeting modulates splicing decisions and if this function is altered in AD. Replicate multivariate analysis of transcript splicing (rMATS) analysis of RNA-Seq data sets from wild-type and AD fly brains revealed a multitude of mammalian-like AS defects. Strikingly, over half of these altered RNAs were bonafide Tip60-RNA targets enriched for in the AD-gene curated database, with some AS alterations prevented against by increasing Tip60 in fly brain. Importantly, human orthologs of several Tip60-modulated spliced genes in Drosophila are well characterized aberrantly spliced genes in human AD brains, implicating disruption of Tip60’s splicing function in AD pathogenesis. Theoretical Importance: The authors' findings support a novel RNA interaction and splicing regulatory function for Tip60 that may underlie AS impairments that hallmark AD etiology. Data Collection: The authors collected data from RNA immunoprecipitation experiments using RNA isolated from 200 pooled wild type Drosophila brains for each of the 3 biological replicates. They also performed genome sequencing (DNBSequencingTM technology, BGI genomics) on 3 replicates for Input RNA and RNA IPs by Tip60. Questions: The question addressed by this study was whether Tip60 has a novel RNA binding/splicing function in human hippocampus and whether this function is impaired in brains from AD fly models and AD patients. Conclusions: The authors' findings support a novel RNA interaction and splicing regulatory function for Tip60 that may underlie AS impairments that hallmark AD etiology.

Keywords: Alzheimer's disease, cognition, aging, neuroepigenetics

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6276 Development and Validation of the University of Mindanao Needs Assessment Scale (UMNAS) for College Students

Authors: Ryan Dale B. Elnar

Abstract:

This study developed a multidimensional need assessment scale for college students called The University of Mindanao Needs Assessment Scale (UMNAS). Although there are context-specific instruments measuring the needs of clinical and non-clinical samples, literature reveals no standardized scales to measure the needs of the college students thus a four-phase item development process was initiated to support its content validity. Comprising seven broad facets namely spiritual-moral, intrapersonal, socio-personal, psycho-emotional, cognitive, physical and sexual, a pyramid model of college needs was deconstructed through FGD sample to support the literature review. Using various construct validity procedures, the model was further tested using a total of 881 Filipino college samples. The result of the study revealed evidences of the reliability and validity of the UMNAS. The reliability indices range from .929-.933. Exploratory and confirmatory factor analyses revealed a one-factor-six-dimensional instrument to measure the needs of the college students. Using multivariate regression analysis, year level and course are found predictors of students’ needs. Content analysis attested the usefulness of the instrument to diagnose students’ personal and academic issues and concerns in conjunction with other measures. The norming process includes 1728 students from the different colleges of the University of Mindanao. Further validation is recommended to establish a national norm for the instrument.

Keywords: needs assessment scale, validity, factor analysis, college students

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6275 Silver Grating for Strong and Reproducible SERS Response

Authors: Y. Kalachyova, O. Lyutakov, V. Svorcik

Abstract:

One of the most significant obstacles for the application of surface enhanced Raman spectroscopy (SERS) is the poor reproducibility of SERS active substrates: SERS intensity can be varied from one substrate to another and moreover along the one substrate surface. High enhancement of the near-field intensity is the key factor for ultrasensitive SERS realization. SERS substrate can be prepared through introduction of highly ordered metal array, where light focusing is achieved through excitation of surface plasmon-polaritons (SPPs). In this work, we report the preparation of silver nanostructures with plasmon absorption peaks tuned by the metal arrangement. Excimer laser modification of poly(methyl methacrylate) followed by silver evaporation is proposed as an effective way for the creation of reproducible and effective surface plasmon-polaritons (SPP)-based SERS substrate. Theoretical and experimental studies were performed to optimize structure parameter for effective SPP excitation. It was found that the narrow range of grating periodicity and metal thickness exist, where SPPs can be most efficiently excited. In spite of the fact, that SERS response was almost always achieved, the enhancement factor was found to vary more with the effectivity of SPP excitation. When the real structure parameters were set to optimal for SPP excitation, a SERS enhancement factor was achieved up to four times. Theoretical and experimental investigation of SPP excitation on the two-dimensional periodical silver array was performed with the aim to make SERS response as high as possible.

Keywords: grating, nanostructures, plasmon-polaritons, SERS

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6274 Restricted Boltzmann Machines and Deep Belief Nets for Market Basket Analysis: Statistical Performance and Managerial Implications

Authors: H. Hruschka

Abstract:

This paper presents the first comparison of the performance of the restricted Boltzmann machine and the deep belief net on binary market basket data relative to binary factor analysis and the two best-known topic models, namely Dirichlet allocation and the correlated topic model. This comparison shows that the restricted Boltzmann machine and the deep belief net are superior to both binary factor analysis and topic models. Managerial implications that differ between the investigated models are treated as well. The restricted Boltzmann machine is defined as joint Boltzmann distribution of hidden variables and observed variables (purchases). It comprises one layer of observed variables and one layer of hidden variables. Note that variables of the same layer are not connected. The comparison also includes deep belief nets with three layers. The first layer is a restricted Boltzmann machine based on category purchases. Hidden variables of the first layer are used as input variables by the second-layer restricted Boltzmann machine which then generates second-layer hidden variables. Finally, in the third layer hidden variables are related to purchases. A public data set is analyzed which contains one month of real-world point-of-sale transactions in a typical local grocery outlet. It consists of 9,835 market baskets referring to 169 product categories. This data set is randomly split into two halves. One half is used for estimation, the other serves as holdout data. Each model is evaluated by the log likelihood for the holdout data. Performance of the topic models is disappointing as the holdout log likelihood of the correlated topic model – which is better than Dirichlet allocation - is lower by more than 25,000 compared to the best binary factor analysis model. On the other hand, binary factor analysis on its own is clearly surpassed by both the restricted Boltzmann machine and the deep belief net whose holdout log likelihoods are higher by more than 23,000. Overall, the deep belief net performs best. We also interpret hidden variables discovered by binary factor analysis, the restricted Boltzmann machine and the deep belief net. Hidden variables characterized by the product categories to which they are related differ strongly between these three models. To derive managerial implications we assess the effect of promoting each category on total basket size, i.e., the number of purchased product categories, due to each category's interdependence with all the other categories. The investigated models lead to very different implications as they disagree about which categories are associated with higher basket size increases due to a promotion. Of course, recommendations based on better performing models should be preferred. The impressive performance advantages of the restricted Boltzmann machine and the deep belief net suggest continuing research by appropriate extensions. To include predictors, especially marketing variables such as price, seems to be an obvious next step. It might also be feasible to take a more detailed perspective by considering purchases of brands instead of purchases of product categories.

Keywords: binary factor analysis, deep belief net, market basket analysis, restricted Boltzmann machine, topic models

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6273 Production of Biodiesel Using Brine Waste as a Heterogeneous Catalyst

Authors: Hilary Rutto, Linda Sibali

Abstract:

In these modern times, we constantly search for new and innovative technologies to lift the burden of our extreme energy demand. The overall purpose of biofuel production research is to source an alternative energy source to replace the normal use of fossil fuel as liquid petroleum products. This experiment looks at the basis of biodiesel production with regards to alternative catalysts that can be used to produce biodiesel. The key factors that will be addressed during the experiments will focus on temperature variation, catalyst additions to the overall reaction, methanol to oil ratio, and the impact of agitation on the reaction. Brine samples sources from nearby plants will be evaluated and tested thoroughly and the key characteristics of these brine samples analysed for the verification of its use as a possible catalyst in biodiesel production. The one factor at a time experimental approach was used in this experiment, and the recycle and reuse characteristics of the heterogeneous catalyst was evaluated.

Keywords: brine sludge, heterogenous catalyst, biodiesel, one factor

Procedia PDF Downloads 171
6272 Identification of Blood Biomarkers Unveiling Early Alzheimer's Disease Diagnosis Through Single-Cell RNA Sequencing Data and Autoencoders

Authors: Hediyeh Talebi, Shokoofeh Ghiam, Changiz Eslahchi

Abstract:

Traditionally, Alzheimer’s disease research has focused on genes with significant fold changes, potentially neglecting subtle but biologically important alterations. Our study introduces an integrative approach that highlights genes crucial to underlying biological processes, regardless of their fold change magnitude. Alzheimer's Single-cell RNA-seq data related to the peripheral blood mononuclear cells (PBMC) was extracted from the Gene Expression Omnibus (GEO). After quality control, normalization, scaling, batch effect correction, and clustering, differentially expressed genes (DEGs) were identified with adjusted p-values less than 0.05. These DEGs were categorized based on cell-type, resulting in four datasets, each corresponding to a distinct cell type. To distinguish between cells from healthy individuals and those with Alzheimer's, an adversarial autoencoder with a classifier was employed. This allowed for the separation of healthy and diseased samples. To identify the most influential genes in this classification, the weight matrices in the network, which includes the encoder and classifier components, were multiplied, and focused on the top 20 genes. The analysis revealed that while some of these genes exhibit a high fold change, others do not. These genes, which may be overlooked by previous methods due to their low fold change, were shown to be significant in our study. The findings highlight the critical role of genes with subtle alterations in diagnosing Alzheimer's disease, a facet frequently overlooked by conventional methods. These genes demonstrate remarkable discriminatory power, underscoring the need to integrate biological relevance with statistical measures in gene prioritization. This integrative approach enhances our understanding of the molecular mechanisms in Alzheimer’s disease and provides a promising direction for identifying potential therapeutic targets.

Keywords: alzheimer's disease, single-cell RNA-seq, neural networks, blood biomarkers

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