Search results for: laboratory molecular diagnostics

Authors: Andres Winston Oreta

Abstract:

Experiential Learning (ELE) is a strategy for enhancing the teaching and learning of courses especially in civil engineering. This paper presents the adaption of the ELE framework in the delivery of various course requirements in an earthquake engineering course. Examples of how ELE is integrated using online tools and hands-on laboratory technology to address the course learning outcomes on earthquake engineering are presented. Student feedback shows that ELE using online tools and technology strengthens students’ understanding and intuition of seismic design and earthquake engineering concepts.

Keywords: earthquake engineering, experiential learning, shake table, online, internet, civil engineering

Procedia PDF Downloads 22

Authors: Mohammad Sadegh Khavarinejad

Abstract:

In this study, genetic diversity of 10 bread wheat genotypes by SSR and RAPD markers was evaluated. 11 primers were used included 6 RAPD primers and 5 SSR primers. RAPDs and SSRs could find 33 and 17 polymorphism respectively. In RAPDs, primers UBC 350 and UBC 109 and in SSRs, Primers Xgwm 469-6D and Xgwm120-2B showed genetic diversity among genotypes more than others.

Keywords: wheat, molecular markers, SSR, RAPD

Procedia PDF Downloads 433

Authors: Khwaja Naweed Seddiqi, Shigeo Honma

Abstract:

Relative permeability is the important parameter controlling the immiscible displacement of multiphase fluids flow in porous medium. The relative permeability for immiscible displacement of two-phase fluids flow (oil and water) in porous medium has been measured in this paper. As a result of the experiment, irreducible water saturation, Swi, residual oil saturation, Sor, and relative permeability curves for Kerosene, Heavy oil and Lubricant oil were determined successfully.

Keywords: relative permeability, two-phase flow, immiscible displacement, porous medium

Procedia PDF Downloads 310

Authors: K. Tunde Olagunju, C. Scott Allen, Freek Van Der Meer

Abstract:

Identification of hydrocarbon oil in remote sensing images is often the first step in monitoring oil during spill events. Most remote sensing methods adopt techniques for hydrocarbon identification to achieve detection in order to model an appropriate cleanup program. Identification on optical sensors does not only allow for detection but also for characterization and quantification. Until recently, in optical remote sensing, quantification and characterization are only potentially possible using high-resolution laboratory and airborne imaging spectrometers (hyperspectral data). Unlike multispectral, hyperspectral data are not freely available, as this data category is mainly obtained via airborne survey at present. In this research, two (2) operational high-resolution multispectral satellites (WorldView-3 and Sentinel-2) are theoretically assessed for their suitability for hydrocarbon characterization, using the hydrocarbon spectral slope model (HYSS). This method utilized the two most persistent hydrocarbon diagnostic/absorption features at 1.73 µm and 2.30 µm for hydrocarbon mapping on multispectral data. In this research, spectra measurement of seven (7) different hydrocarbon oils (crude and refined oil) taken on ten (10) different substrates with the use of laboratory ASD Fieldspec were convolved to Sentinel-2 and WorldView-3 resolution, using their full width half maximum (FWHM) parameter. The resulting hydrocarbon slope values obtained from the studied samples enable clear qualitative discrimination of most hydrocarbons, despite the presence of different background substrates, particularly on WorldView-3. Due to close conformity of central wavelengths and narrow bandwidths to key hydrocarbon bands used in HYSS, the statistical significance for qualitative analysis on WorldView-3 sensors for all studied hydrocarbon oil returned with 95% confidence level (P-value ˂ 0.01), except for Diesel. Using multifactor analysis of variance (MANOVA), the discriminating power of HYSS is statistically significant for most hydrocarbon-substrate combinations on Sentinel-2 and WorldView-3 FWHM, revealing the potential of these two operational multispectral sensors as rapid response tools for hydrocarbon mapping. One notable exception is highly transmissive hydrocarbons on Sentinel-2 data due to the non-conformity of spectral bands with key hydrocarbon absorptions and the relatively coarse bandwidth (> 100 nm).

Keywords: hydrocarbon, oil spill, remote sensing, hyperspectral, multispectral, hydrocarbon-substrate combination, Sentinel-2, WorldView-3

Procedia PDF Downloads 216

Authors: John Masani Nduko, Joseph Wafula Matofari

Abstract:

Many spontaneously fermented milk products are produced in Kenya, where they are integral to the human diet and play a central role in enhancing food security and income generation via small-scale enterprises. Fermentation enhances product properties such as taste, aroma, shelf-life, safety, texture, and nutritional value. Some of these products have demonstrated therapeutic and probiotic effects although recent reports have linked some to death, biotoxin infections, and esophageal cancer. These products are mostly processed from poor quality raw materials under unhygienic conditions resulting to inconsistent product quality and limited shelf-lives. Though very popular, research on their processing technologies is low, and none of the products has been produced under controlled conditions using starter cultures. To modernize the processing technologies for these products, our study aims at describing the microbiology and biochemistry of a representative Kenyan spontaneously fermented milk product, Mursik using modern biotechnology (DNA sequencing) and their chemical composition. Moreover, co-creation processes reflecting stakeholders’ experiences on traditional fermented milk production technologies and utilization, ideals and senses of value, which will allow the generation of products based on common ground for rapid progress will be discussed. Knowledge of the value of clean starting raw material will be emphasized, the need for the definition of fermentation parameters highlighted, and standard equipment employment to attain controlled fermentation discussed. This presentation will review the available information regarding traditional fermented milk (Mursik) and highlight our current research work on the application of molecular approaches (metagenomics) for the valorization of Mursik production process through starter culture/ probiotic strains isolation and identification, and quality and safety aspects of the product. The importance of the research and future research areas on the same subject will also be highlighted.

Keywords: lactic acid bacteria, high throughput biotechnology, spontaneous fermentation, Mursik

Procedia PDF Downloads 293

Authors: Judith Hanke

Abstract:

Due to the increase of students with reading difficulties, digital reading support with diagnostics was developed to foster the individual student's reading comprehension. The digital reading support focused on the reading comprehension of elementary school students. The digital reading packages consist of literary texts with aligned reading exercises. The number of students with German as a second language is growing in Germany. Students with multilingualism, language 1, and language 2 learn German together in school. The research's focus is on determining whether and to what extent multilingualism, language 1, and language 2 affect reading comprehension. For the methodology, an ABA design was selected for the intervention study to examine the reading support. The study was expedited from April 2023 until July 2023 and collected quantitative data of individuals, groups, and classes. It comprised a survey group (N = 58) and a control group (N = 53). The quantitative data was collected from 3 classes of 3 teachers and 47 students for all three test times. To show differences between the groups, a standardized reading comprehension test was used for the three test times, pretest, posttest, and follow-up. The standardized test consists of three subtests regarding word comprehension, sentence comprehension, and text comprehension. The main findings include that students who spoke German as their first language had the best test scores. Interestingly, students with a different language had better testing scores than students with German as the first language and (an) other language/s. Also, the students with another language outperformed the native language speakers in one of the subtests of the post-testing. The variables of spoken language at home and German as a second language were also examined and correlated with the test results. One significant correlation was found between spoken language at home and the text comprehension test of the pretesting. Additionally, the variable German as a second language had multiple significant correlations in the pretest, posttest and follow-up. The study's significance is to understand the influence of several languages, language 1, and language 2, on reading comprehension.

Keywords: multilingualism, language 1, language 2, reading comprehension, second language

Procedia PDF Downloads 29

Authors: Japhet N. Mwambusi

Abstract:

High tropical forests deforestation for solid wood furniture industry is among of climate change contributing agents. This pressure indirectly is caused by furniture joints failure due to poor gluing technology based on improper use of different glues to different wood species which lead to low quality and weak wood-glue joints. This study was carried in order to run performance tests of wood glues on different wood species used in wood workshops: Morogoro Tanzania whereby three popular wood species of C. lusitanica, T. glandis and E. maidenii were tested against five glues of Woodfix, Bullbond, Ponal, Fevicol and Coral found in the market. The findings were necessary on developing a guideline for proper glue selection for a particular wood species joining. Random sampling was employed to interview carpenters while conducting a survey on the background of carpenters like their education level and to determine factors that influence their glues choice. Monsanto Tensiometer was used to determine bonding strength of identified wood glues to different wood species in use under British Standard of testing wood shear strength (BS EN 205) procedures. Data obtained from interviewing carpenters were analyzed through Statistical Package of Social Science software (SPSS) to allow the comparison of different data while laboratory data were compiled, related and compared by the use of MS Excel worksheet software as well as Analysis of Variance (ANOVA). Results revealed that among all five wood glues tested in the laboratory to three different wood species, Coral performed much better with the average shear strength 4.18 N/mm², 3.23 N/mm² and 5.42 N/mm² for Cypress, Teak and Eucalyptus respectively. This displays that for a strong joint to be formed to all tree wood species for soft wood and hard wood, Coral has a first priority in use. The developed table of guideline from this research can be useful to carpenters on proper glue selection to a particular wood species so as to meet glue-bond strength. This will secure furniture market as well as reduce pressure to the forests for furniture production because of the strong existing furniture due to their strong joints. Indeed, this can be a good strategy on reducing climate change speed in tropics which result from high deforestation of trees for furniture production.

Keywords: climate change, deforestation, gluing technology, joint failure, wood-glue, wood species

Procedia PDF Downloads 240

Authors: E. Syngelaki, C. C. F. Schinkel, S. Klatt, E. Hörandl

Abstract:

Environmental influence can alter the conditions for plant development and can trigger changes in epigenetic variation. Thus, the exposure to abiotic environmental stress can lead to different DNA methylation profiles and may have evolutionary consequences for adaptation. Epigenetic control mechanisms may further influence mode of reproduction. The alpine species R. kuepferi has diploid and tetraploid cytotypes, that are mostly sexual and facultative apomicts, respectively. Hence, it is a suitable model system for studying the correlations of mode of reproduction, ploidy, and environmental stress. Diploid and tetraploid individuals were placed in two climate chambers and treated with low (+7°C day/+2°C night, -1°C cold shocks for three nights per week) and warm (control) temperatures (+15°C day/+10°C night). Subsequently, methylation sensitive-Amplified Fragment-Length Polymorphism (AFPL) markers were used to screen genome-wide methylation alterations triggered by stress treatments. The dataset was analyzed for four groups regarding treatment (cold/warm) and ploidy level (diploid/tetraploid), and also separately for full methylated, hemi-methylated and unmethylated sites. Patterns of epigenetic variation suggested that diploids differed significantly in their profiles from tetraploids independent from treatment, while treatments did not differ significantly within cytotypes. Furthermore, diploids are more differentiated than the tetraploids in overall methylation profiles of both treatments. This observation is in accordance with the increased frequency of apomictic seed formation in diploids and maintenance of facultative apomixis in tetraploids during the experiment. Global analysis of molecular variance showed higher epigenetic variation within groups than among them, while locus-by-locus analysis of molecular variance showed a high number (54.7%) of significantly differentiated un-methylated loci. To summarise, epigenetic variation seems to depend on ploidy level, and in diploids may be correlated to changes in mode of reproduction. However, further studies are needed to elucidate the mechanism and possible functional significance of these correlations.

Keywords: apomixis, cold stress, DNA methylation, Ranunculus kuepferi

Procedia PDF Downloads 160

Authors: Lubomir Machuca, Vit Fara

Abstract:

Ammonium nitrate (AN) is produced by the reaction of ammonia and nitric acid, and a waste condensate is obtained. The condensate contains pure AN in concentration up to 10g/L. The salt content in the condensate is too high to discharge immediately into the river thus it must be treated. This study is concerned with the treatment of condensates from an industrial AN production by combination of electrodialysis (ED) and electrodeionization (EDI). The condensate concentration was in range 1.9–2.5g/L of AN. A pilot ED module with 25 membrane pairs following by a laboratory EDI module with 10 membrane pairs operated continuously during 800 hours. Results confirmed that the combination of ED and EDI is suitable for the condensate treatment.

Keywords: desalination, electrodialysis, electrodeionization, fertilizer industry

Procedia PDF Downloads 240

Authors: Ayman K. El Essawy, Amal M. Hosny, Hala M. Abu Shady

Abstract:

The rapid detection of TB and drug resistance, both optimizes treatment and improves outcomes. In the current study, respiratory specimens were collected from 155 patients. Conventional susceptibility testing and MIC determination were performed for rifampicin (RIF) and isoniazid (INH). Genotype MTBDRplus assay, which is a molecular genetic assay based on the DNA-STRIP technology and specific gene sequencing with primers for rpoB, KatG, and mab-inhA genes were used to detect mutations associated with resistance to rifampicin and isoniazid. In comparison to other categories, most of rifampicin resistant (61.5%) and isoniazid resistant isolates (47.1%) were from patients relapsed in treatment. The genotypic profile (using Genotype MTBDRplus assay) of multi-drug resistant (MDR) isolates showed missing of katG wild type 1 (WT1) band and appearance of mutation band katG MUT2. For isoniazid mono-resistant isolates, 80% showed katG MUT1, 20% showed katG MUT1, and inhA MUT1, 20% showed only inhA MUT1. Accordingly, 100% of isoniazid resistant strains were detected by this assay. Out of 17 resistant strains, 16 had mutation bands for katG distinguished high resistance to isoniazid. The assay could clearly detect rifampicin resistance among 66.7% of MDR isolates that showed mutation band rpoB MUT3 while 33.3% of them were considered as unknown. One mono-resistant rifampicin isolate did not show rifampicin mutation bands by Genotype MTBDRplus assay, but it showed an unexpected mutation in Codon 531 of rpoB by DNA sequence analysis. Rifampicin resistance in this strain could be associated with a mutation in codon 531 of rpoB (based on molecular sequencing), and Genotype MTBDRplus assay could not detect the associated mutation. If the results of Genotype MTBDRplus assay and sequencing were combined, this strain shows hetero-resistance pattern. Gene sequencing of eight selected isolates, previously tested by Genotype MTBDRplus assay, could detect resistance mutations mainly in codon 315 (katG gene), position -15 in inhA promotes gene for isoniazid resistance and codon 531 (rpoB gene) for rifampicin resistance. Genotyping techniques allow distinguishing between recurrent cases of reinfection or reactivation and supports epidemiological studies.

Keywords: M. tuberculosis, rpoB, KatG, inhA, genotype MTBDRplus

Procedia PDF Downloads 167

Authors: Faijal Ali, Darga Kumar N., Ravikant Singh, Rajnil Lal

Abstract:

Fiji has three major landforms such as plains, low mountains, and hills. The low land soils are formed on beach sand. Fiji soils contain high concentration of iron (III), aluminum oxides and hydroxides. The soil possesses reddish or yellowish colour. The characterization of lateritic soils collected from different locations along the national highway in Viti Levu, Fiji Islands. The research has been carried out mainly to understand the physical and strength properties to assess their suitability for the highway and building construction. In this paper, the field tests such as dynamic cone penetrometer test, field vane shear, field density and laboratory tests such as unconfined compression stress, compaction, grain size analysis and Atterberg limits are conducted. The test results are analyzed and presented. From the results, it is revealed that the soils are having more percentage of silt and clay which is more than 80% and 5 to 15% of fine to medium sand is noticed. The dynamic cone penetrometer results up to 3m depth had similar penetration resistance. For the first 1m depth, the rate of penetration is found 300mm per 3 to 4 blows. In all the sites it is further noticed that the rate of penetration at depths beyond 1.5 m is decreasing for the same number of blows as compared to the top soil. From the penetration resistance measured through dynamic cone penetrometer test, the California bearing ratio and allowable bearing capacities are 4 to 5% and 50 to 100 kPa for the top 1m layer and below 1m these values are increasing. The California bearing ratio of these soils for below 1m depth is in the order of 10% to 20%. The safe bearing capacity of these soils below 1m and up to 3m depth is varying from 150 kPa to 250 kPa. The field vane shear was measured within a depth of 1m from the surface and the values were almost similar varying from 60 kPa to 120 kPa. The liquid limit and plastic limits of these soils are in the range of 40 to 60% and 20 to 25%. Overall it is found that the top 1m soil along the national highway in majority places possess a soft to medium stiff behavior with low to medium bearing capacity as well low California bearing ratio values. It is recommended to ascertain these soils behavior in terms of geotechnical parameters before taking up any construction activity.

Keywords: California bearing ratio, dynamic cone penetrometer test, field vane shear, unconfined compression stress.

Procedia PDF Downloads 186

Authors: A. Berraf-Tebbal, Z. Bouznad, , A.J.L. Phillips

Abstract:

Phaeomoniella is a fungus genus in the mitosporic ascomycota which includes Phaeomoniella chlamydospora specie associated with two declining diseases on grapevine (Vitis vinifera) namely Petri disease and esca. Recent studies have shown that several Phaeomoniella species also cause disease on many other woody crops, such as forest trees and woody ornamentals. Two new species, Phaeomoniella zymoides and Phaeomoniella pinifoliorum H.B. Lee, J.Y. Park, R.C. Summerbell et H.S. Jung, were isolated from the needle surface of Pinus densiflora Sieb. et Zucc. in Korea. The identification of species in Phaeomoniella genus can be a difficult task if based solely on morphological and cultural characters. In this respect, the application of molecular methods, particularly PCR-based techniques, may provide an important contribution. MSP-PCR (microsatellite primed-PCR) fingerprinting has proven useful in the molecular typing of fungal strains. The high discriminatory potential of this method is particularly useful when dealing with closely related or cryptic species. In the present study, the application of PCR fingerprinting was performed using the micro satellite primer M13 for the purpose of species identification and strain typing of 84 Phaeomoniella -like isolates collected from grapevines with typical symptoms of dieback. The bands produced by MSP-PCR profiles divided the strains into 3 clusters and 5 singletons with a reproducibility level of 80%. Representative isolates from each group and, when possible, isolates from Eutypa dieback and esca symptoms were selected for sequencing of the ITS region. The ITS sequences for the 16 isolates selected from the MSP-PCR profiles were combined and aligned with sequences of 18 isolates retrieved from GenBank, representing a selection of all known Phaeomoniella species. DNA sequences were compared with those available in GenBank using Neighbor-joining (NJ) and Maximum-parsimony (MP) analyses. The phylogenetic trees of the ITS region revealed that the Phaeomoniella isolates clustered with Phaeomoniella chlamydospora reference sequences with a bootstrap support of 100 %. The complexity of the pathosystems vine-trunk diseases shows clearly the need to identify unambiguously the fungal component in order to allow a better understanding of the etiology of these diseases and justify the establishment of control strategies against these fungal agents.

Keywords: Genotyping, MSP-PCR, ITS, phylogeny, trunk diseases

Procedia PDF Downloads 480

Authors: M. Velana, G. Rinkenauer

Abstract:

Nowadays, nurses are facing unprecedented amounts of pressure due to the ongoing global health demands. Work-related stress can cause a high physical and psychological workload, which can lead, in turn, to burnout. On the physiological level, stress triggers an initial activation of the sympathetic nervous and adrenomedullary systems resulting in increases in cardiac activity. Furthermore, activation of the hypothalamus-pituitary-adrenal axis provokes endocrine and immune changes leading to the release of cortisol and cytokines in an effort to re-establish body balance. Based on the current state of the literature, it has been identified that resilience and mindfulness exercises among nurses can effectively decrease stress and improve mood. However, it is still unknown what relaxation techniques would be suitable for and to what extent would be effective to decrease psychophysiological arousal deriving from either a physiological or a psychological stressor. Moreover, although cardiac activity and cortisol are promising candidates to examine the effectiveness of relaxation to reduce stress, it still remains to shed light on the role of cytokines in this process so as to thoroughly understand the body’s response to stress and to relaxation. Therefore, the main aim of the present study is to develop a comprehensive experimental paradigm and assess different relaxation techniques, namely progressive muscle relaxation and a mindfulness exercise originating from cognitive therapy by means of biofeedback, under highly controlled laboratory conditions. An experimental between-subject design will be employed, where 120 participants will be randomized either to a physiological or a psychological stress-related experiment. Particularly, the cold pressor test refers to a procedure in which the participants have to immerse their non-dominant hands into ice water (2-3 °C) for 3 min. The participants are requested to keep their hands in the water throughout the whole duration. However, they can immediately terminate the test in case it would be barely tolerable. A pre-test anticipation phase and a post-stress period of 3 min, respectively, are planned. The Trier Social Stress Test will be employed to induce psychological stress. During this laboratory stressor, the participants are instructed to give a 5-min speech in front of a committee of communication specialists. Before the main task, there is a 10-min anticipation period. Subsequently, participants are requested to perform an unexpected arithmetic task. After stress exposure, the participants will perform one of the relaxation exercises (treatment condition) or watch a neutral video (control condition). Electrocardiography, salivary samples, and self-report will be collected at different time points. The preliminary results deriving from the pilot study showed that the aforementioned paradigm could effectively induce stress reactions and that relaxation might decrease the impact of stress exposure. It is of utmost importance to assess how the human body responds under different stressors and relaxation exercises so that an evidence-based intervention could be transferred in a clinical setting to improve nurses’ general health. Based on suggestive future laboratory findings, the research group plans to conduct a pilot-level randomized study to decrease stress and promote well-being among nurses who work in the stress-riddled environment of a hospital located in Northern Germany.

Keywords: nurses, psychophysiology, relaxation, stress

Procedia PDF Downloads 110

Authors: Murat Iskender Aktaş, Sadi Ergin, Rasime Acar Aktaş

Abstract:

In the light of fast-paced changes and developments in the health sector, the Ministry of Health started a new structuring with decree law numbered 663 within the scope of the Project of Transformation in Health. Accordingly, hospitals should ensure patient satisfaction through more efficient, more effective use of resources and sustainable finance by placing patients in the centre and should operate to increase efficiency to its maximum level while doing these. Within this study, in order to find out how efficient the hospitals were in terms of medical waste management between the years 2011-2014, the data from six hospitals of Amasya Public Hospitals Union were evaluated separately through Data Envelopment Analysis (DEA) method. First of all, input variables were determined. Input variables were the number of patients admitted to polyclinics, the number of inpatients in clinics, the number of patients who were operated and the number of patients who applied to the laboratory. Output variable was the cost of medical wastes in Turkish liras. Each hospital’s total medical waste level before and after public hospitals union; the amounts of average medical waste per patient admitted to polyclinics, per inpatient in clinics, per patient admitted to laboratory and per operated patient were compared within each group. In addition, average medical waste levels and costs were compared for Turkey in general and Europe in general. Paired samples t-test was used to find out whether the changes (increase-decrease) after public hospitals union were statistically significant. The health facilities that were unsuccessful in terms of medical waste management before and after public hospital union and the factors that caused this failure were determined. Based on the results, for each health facility that was ineffective in terms of medical waste management, the level of improvement required for each input was determined. The results of the study showed that there was an improvement in medical waste management applications after the health facilities became a member of public hospitals union; their medical waste levels were lower than the average of Turkey and Europe while the averages of cost of disposal were the highest.

Keywords: medical waste management, cost of medical waste, public hospitals, data envelopment analysis

Procedia PDF Downloads 415

Authors: Ardak Myrzabekova

Abstract:

Sentinel epidemiological surveillance (SES) of severe acute respiratory infections (SARI) was introduced in the Republic of Kazakhstan in 2008. The purpose of this study was to analyze SES of flu among SARI patients in the Republic of Kazakhstan during last two flu seasons. Comparative analysis was conducted of SARI morbidity during 40 – 23 weeks of 2014/2015 (season 2014) and 2015/2016 (season 2015) in online base (http:\\ses.dec.kz). In the database during season 2014 were 1,398 SARI patients and 1,985 patients during season 2015. Individual data (clinical, epidemiological and laboratory) of SARI cases were collected based on the questionnaire and were put into the flu electronic system. The studied population was residents of the Republic of Kazakhstan who addressed for medical help in 24 sentinel in-patient clinics in 9 sentinel regions of the country. Swabs from nose and throat were taken for laboratory testing from SARI patients who met the standard case definition. The samples were examined in virology labs of sentinel regions using PCR and 'AmpliSens' test systems made in Russia. The first positive results for flu during season 2014 were obtained on 48 week, during season 2015 – on 46 week. The increase of the number of hospitalized SARI patients was observed during 42 week of 2015 – 01 week of 2016, and during 03 - 06 weeks of 2016, with fluctuating SARI incidence rate from 171 to 444 per 1,000 hospitalized. The highest SARI incidence rate during season 2014 were observed during 01 - 03 weeks of 2015: from 389 to 466 per 1,000 hospitalized. Patients admitted to the ICU during season 2015 were 3.0% (60) SARI patients, compared to 2.7% (38) in 2014 (p=0.3), obtaining oxygen therapy 1.0% (21) compared to 0.3% (5), accordingly, (р=0.009); with shortness of breath 74.8% (1,486) compared to 72.6% (1,015), (р=0.07); with impairment of consciousness 1.0% (21) compared to 0.6% (9), (р=0.11); with muscle pain 19.3% (384) compared to 13.6% (191), (р < 0.001); with joint pain 13.3% (265) compared to 9.3% (131), (p < 0.001). During season 2015 the prevailing subtype of flu А was А/Н1N1-09, it was observed mainly in the age group 30-64: 32.5% (169/520). During season 2014 flu А/Н3N2 was observed mainly in the age group 15-29: 43.6% (106/243). Among children under 14 flu А/Н1N1-09 during season 2015 was 37.3% (194/520), during season 2014 flu А/Н3N2 – 34.9% (85/243). Earlier beginning of the flu season was noted in 2015-2016 and a longer period of hospitalization of SARI patients, with high SARI morbidity rates, unlike season 2014-2015. Season 2015-2016 was characterized by prevailing circulation of virus of flu А/Н1N1-09, mainly in the age group 30-64, and also among children under 14. During season 2014-2015 the virus circulating in the country was А/Н3N2, which was observed mainly in the age group 15-29 and among children under 14.

Keywords: flu, electronic system, sentinel epidemiological surveillance, severe acute respiratory infections

Procedia PDF Downloads 227

Authors: Itji Diana Daud, Nuniek Widiayani

Abstract:

The fungus Beauveria bassiana can be endophytic in maize. The fungus was recovered in culture from stems, leaves and roots after a month planting. This phenomenon was shown until the third generation of the corn. The result from laboratory shows that B. bassiana appear in F1, F2 and F3 in order 70, 80 and 90% in the roots, 80% in the stems in all generation, 90, 80 and 70% in leaves. In CFU’s ml-1 of B. bassiana in corn seed, show F1 was 8.9 x 106, F2 was 8.1 x 106 and F3 was 7.8 x 106. The research showed that B. Bassiana as endophyte still remain to the third generation. Innovation to the corn seed which is endophyte seed is essential to protect from the attack of corn borer and to avoid the usage of insecticide.

Keywords: endophytic, recovered, third generation, Beauveria bassiana

Procedia PDF Downloads 283

Authors: Muhammad A. Ashraf, Scott Steinmetz, Matthew J. Dunn, Assaad R. Masri

Abstract:

This study focuses on the evolution of soot and soot precursors in three different piloted diffusion turbulent flames. The fuel composition is as follow flame A (ethylene/nitrogen, 2:3 by volume), flame B (ethylene/air, 2:3 by volume), and flame C (pure methane). These flames are stabilized using a 4mm diameter jet surrounded by a pilot annulus with an outer diameter of 15 mm. The pilot issues combustion products from stoichiometric premixed flames of hydrogen, acetylene, and air. In all cases, the jet Reynolds number is 10,000, and air flows in the coflow stream at a velocity of 5 m/s. Time-resolved laser-induced fluorescence (LIF) is collected at two wavelength bands in the visible (445 nm) and UV regions (266 nm) along with laser-induced incandescence (LII). The combined results are employed to study concentration, size, and growth of soot and precursors. A set of four fast photo-multiplier tubes are used to record emission data in temporal domain. A 266nm laser pulse preferentially excites smaller nanoparticles which emit a fluorescence spectrum which is analysed to track the presence, evolution, and destruction of nanoparticles. A 1064nm laser pulse excites sufficiently large soot particles, and the resulting incandescence is collected at 1064nm. At downstream and outer radial locations, intermittency becomes a relevant factor. Therefore, data collected in turbulent flames is conditioned to account for intermittency so that the resulting mean profiles for scattering, fluorescence, and incandescence are shown for the events that contain traces of soot. It is found that in the upstream regions of the ethylene-air and ethylene-nitrogen flames, the presence of soot precursors is rather similar. However, further downstream, soot concentration grows larger in the ethylene-air flames.

Keywords: laser induced incandescence, laser induced fluorescence, soot, nanoparticles

Procedia PDF Downloads 146

Authors: Siddharth Jain, Abhenil Mittal, Surendra Kumar Sharma

Abstract:

Background: With its recent emergence and re-emergence, dengue has become a major international public health concern, imposing significant financial burden especially in developing countries. Despite aggressive control measures in place, India experienced one of its largest outbreaks in 2015 with Delhi being most severely affected. There is a lack of reliable predictors of disease severity and mortality in dengue. The present study was carried out to identify these predictors during the 2015 outbreak. Methods: This prospective observational study conducted at an apex tertiary care center in Delhi, India included confirmed adult dengue patients admitted between August-November 2015. Patient demographics, clinical details, and laboratory findings were recorded in a predesigned proforma. Appropriate statistical tests were used to summarize and compare the clinical and laboratory characteristics and derive predictors of mortality and severe disease, while developing a clinical risk score for mortality. Serotype analysis was also done for 75 representative samples to identify the dominant serotypes. Results: Data of 369 patients were analyzed (mean age 30.9 years; 67% males). Of these, 198 (54%) patients had dengue fever, 125 (34%) had dengue hemorrhagic fever (DHF Grade 1,2)and 46 (12%) developed dengue shock syndrome (DSS). Twenty two (6%) patients died. Late presentation to the hospital (≥5 days after onset) and dyspnoea at rest were identified as independent predictors of severe disease. Age ≥ 24 years, dyspnoea at rest and altered sensorium were identified as independent predictors of mortality. A clinical risk score was developed (12*age + 14*sensorium + 10*dyspnoea) which, if ≥ 22, predicted mortality with a high sensitivity (81.8%) and specificity (79.2%). The predominant serotypes in Delhi (2015) were DENV-2 and DENV-4. Conclusion: Age ≥ 24 years, dyspnoea at rest and altered sensorium were identified as independent predictors of mortality. Platelet counts did not determine the outcome in dengue patients. Timely referral/access to health care is important. Development and use of validated predictors of disease severity and simple clinical risk scores, which can be applied in all healthcare settings, can help minimize mortality and morbidity, especially in resource limited settings.

Keywords: dengue, mortality, predictors, severity

Procedia PDF Downloads 307

Authors: Marius Sebastian Secula, Andreea Vajda, Benoit Cagnon, Ioan Mamaliga

Abstract:

One of the most important classes of pollutants is represented by dyes. The synthetic character and complex molecular structure make them more stable and difficult to be biodegraded in water. The treatment of wastewaters containing dyes in order to separate/degrade dyes is of major importance. Various techniques have been employed to remove and/or degrade dyes in water. Advanced oxidation processes (AOPs) are known as among the most efficient ones towards dye degradation. The aim of this work is to investigate the efficiency of a cheap Iron-impregnated activated carbon Fenton-like catalyst in order to degrade organic compounds in aqueous solutions. In the presented study an anionic dye, Indigo Carmine, is considered as a model pollutant. Various AOPs are evaluated for the degradation of Indigo Carmine to establish the effect of the prepared catalyst. It was found that the Iron(II)-embedded activated carbon composite enhances significantly the degradation process of Indigo Carmine. Using the wet impregnation procedure, 5 g of L27 AC material were contacted with Fe(II) solutions of FeSO4 precursor at a theoretical iron content in the resulted composite of 1 %. The L27 AC was impregnated for 3h at 45°C, then filtered, washed several times with water and ethanol and dried at 55 °C for 24 h. Thermogravimetric analysis, Fourier transform infrared, X-ray diffraction, and transmission electron microscopy were employed to investigate the structural, textural, and micromorphology of the catalyst. Total iron content in the obtained composites and iron leakage were determined by spectrophotometric method using phenantroline. Photo-catalytic tests were performed using an UV - Consulting Peschl Laboratory Reactor System. UV light irradiation tests were carried out to determine the performance of the prepared Iron-impregnated composite towards the degradation of Indigo Carmine in aqueous solution using different conditions (17 W UV lamps, with and without in-situ generation of O3; different concentrations of H2O2, different initial concentrations of Indigo Carmine, different values of pH, different doses of NH4-OH enhancer). The photocatalytic tests were performed after the adsorption equilibrium has been established. The obtained results emphasize an enhancement of Indigo Carmine degradation in case of the heterogeneous photo-Fenton process conducted with an O3 generating UV lamp in the presence of hydrogen peroxide. The investigated process obeys the pseudo-first order kinetics. The photo-Fenton degradation of IC was tested at different values of initial concentration. The obtained results emphasize an enhancement of Indigo Carmine degradation in case of the heterogeneous photo-Fenton process conducted with an O3 generating UV lamp in the presence of hydrogen peroxide. Acknowledgments: This work was supported by a grant of the Romanian National Authority for Scientific Research and Innovation, CNCS - UEFISCDI, project number PN-II-RU-TE-2014-4-0405.

Keywords: photodegradation, heterogeneous Fenton, anionic dye, carbonaceous composite, screening factorial design

Procedia PDF Downloads 257

Authors: Harithas Aruchchunan

Abstract:

Silage production is an essential aspect of dairy farming, used to provide high-quality feed to ruminants. However, dairy farmers in Northern Province Sri Lanka are facing multiple challenges that compromise the quality and quantity of silage produced. To tackle these challenges, promoting silage feeding has become an essential component of sustainable dairy farming practices. In this study, silage bale samples were collected from a newly started silage baling factory in Jaffna, Northern province and their quality was analysed at the Veterinary Research Institute laboratory in Kandy in March 2023. The results show the nutritional composition of three Napier grass cultivars: Super Napier, CO6, and Indian Red Napier (BH18). The main parameters analysed were dry matter, pH, lactic acid, soluble carbohydrate, ammonia nitrogen, ash, crude protein, NDF, and ADF. The results indicate that Super Napier and CO6 have higher crude protein content and lower ADF levels, making them suitable for producing high-quality silage. The pH levels of all three cultivars were safe, and the ammonia nitrogen levels were considered appropriate. However, laboratory results indicate that the quality of silage bales produced can be further enhanced. Dairy farmers should be encouraged to adopt these cultivars to achieve better yields as they are high in protein and are better suited to Northern Province's soil and climate. Therefore, it is vital to educate small-scale fodder producers, who supply the raw material to silage factories, on the best practices of cultivating these new cultivars. To improve silage bale production and quality in Northern Province Sri Lanka, we recommend increasing public awareness about silage feeding, providing education and training to dairy farmers and small-scale fodder producers on modern silage production techniques and improving the availability of raw materials for silage production. Additionally, Napier grass cultivars need to be promoted among dairy farmers for better production and quality of silage bales. Failing to improve the quality and quantity of silage bale production could not only lead to the decline of dairy farming in Northern Province Sri Lanka but also the negative impact on the economy

Keywords: silage bales, dairy farming, economic crisis, Sri Lanka

Procedia PDF Downloads 92

Authors: Tuiana A. Aiusheeva, Sergey S. Tamozhnikov, Alexander E. Saprygin, Arina A. Antonenko, Valentina V. Stepanova, Natalia N. Tolstykh, Alexander N. Savostyanov

Abstract:

Situation of choice is an important condition for the formation of essential character qualities of a child, such as being initiative, responsible, hard-working. We have studied the behavioral and EEG reactions in Russian schoolchildren during recognition of syntactic errors in emotionally colored sentences that describe the choice situation. Twenty healthy children (mean age 9,0±0,3 years, 12 boys, 8 girls) were examined. Forty sentences were selected for the experiment; the half of them contained a syntactic error. The experiment additionally had the hidden condition: 50% of the sentences described the children's own choice and were emotionally colored (positive or negative). The other 50% of the sentences described the forced-choice situation, also with positive or negative coloring. EEG were recorded during execution of error-recognition task. Reaction time and quality of syntactic error detection were chosen as behavioral measures. Event-related spectral perturbation (ERSP) was applied to characterize the oscillatory brain activity of children. There were two time-frequency intervals in EEG reactions: (1) 500-800 ms in the 3-7 Hz frequency range (theta synchronization) and (2) 500-1000 ms in the 8-12 Hz range (alpha desynchronization). We found out that behavioral and brain reactions in child brain during recognition of positive and negative sentences describing forced-choice situation did not have significant differences. Theta synchronization and alpha desynchronization were stronger during recognition of sentences with children's own choice, especially with negative coloring. Also, the quality and execution time of the task were higher for this types of sentences. The results of our study will be useful for improvement of teaching methods and diagnostics of children affective disorders.

Keywords: choice situation, electroencephalogram (EEG), emotionally colored sentences, schoolchildren

Procedia PDF Downloads 269

Authors: Theresa Bacon-Baguley, Martina Reinhold

Abstract:

Introduction: Evaluation and analysis are the cornerstones of any successful program in higher education. When developing a program at a distant campus, it is essential that the process of evaluation and analysis be orchestrated in a timely manner with tools that can identify both the positive and negative components of distant education. We describe the utilization of a newly developed tool used to evaluate and analyze the successful expansion to a distant campus using Telepresence Technology. Like interactive television, Telepresence allows live interactive delivery but utilizes broadband cable. The tool developed is adaptable to any distant campus as the framework for the tool was derived from a systematic review of the literature. Methodology: Because Telepresence is a relatively new delivery system, the evaluation tool was developed based on a systematic review of literature in the area of distant education and ITV. The literature review identified four potential areas of concern: 1) technology, 2) confidence in the system, 3) faculty delivery of the content and, 4) resources at each site. Each of the four areas included multiple sub-components. Benchmark values were determined to be 80% or greater positive responses to each of the four areas and the individual sub-components. The tool was administered each semester during the didactic phase of the curriculum. Results: Data obtained identified site-specific issues (i.e., technology access, student engagement, laboratory access, and resources), as well as issues common at both sites (i.e., projection screen size). More specifically, students at the parent location did not have adequate access to printers or laboratory space, and students at the distant campus did not have adequate access to library resources. The evaluation tool identified that both sites requested larger screens for visualization of the faculty. The deficiencies were addressed by replacing printers, including additional orientation for students on library resources and increasing the screen size of the Telepresence system. When analyzed over time, the issues identified in the tool as deficiencies were resolved. Conclusions: Utilizing the tool allowed adjustments of the Telepresence delivery system in a timely manner resulting in successful implementation of an entire curriculum at a distant campus.

Keywords: physician assistant, telepresence technology, distant education, assessment

Procedia PDF Downloads 124

Authors: M. Naresh, Pratik Chaturvedi, Srishti Yadav, Varun Dutt, K. V. Uday

Abstract:

Changes in the Earth’s climate are likely to increase natural hazards such as drought, floods, earthquakes, landslides, etc. The present study focusing on to early warning systems (EWS) of landslides, major issues in Himalayan region without prominence to deforestation, encroachments and un-engineered cutting of slopes and reforming for infrastructural purposes. EWS can be depicted by conducting a series of flume tests using micro-electro mechanical systems sensors data after reaching threshold values under controlled laboratory conditions. Based on the threshold value database, an alert will be sent via SMS.

Keywords: slope-instability, flume test, sensors, early warning system

Procedia PDF Downloads 266

Authors: M. Dekmouche, M. Hadjada, Z. Rahmani, M. Saidi

Abstract:

The effect of iodide ions on the corrosion inhibition of mild steel in 20% sulfuric acid in the presence of 3-méthylthio-5-p-méthoxyphényl-1,2-dithiolylium against anion (I-) A1 synthesized in our laboratory,was studied by different electrochemical techniques such as electrochemical impedance spectroscopy, potentiodynamic polarization. The obtained results showed that A1 effectively reduces the corrosion rate of steel. The adsorption of 3-méthylthio-5-p-méthoxyphényl-1,2-dithiolylium against anion (I-) followed Langmuir and temkin adsorption isotherm.

Keywords: steel XC52, corrosion, inhibition, 3-méthylthio-5-p-méthoxyphényl-1, 2-dithiolylium against anion (I-) , sulfuric acid

Procedia PDF Downloads 328

Authors: Gurbanova J., Majidova N., Ali-Zade S., Hasanova A., Mikailzade P.

Abstract:

Currently, the problem of oncogynecological diseases is widespread and remains relevant in terms of quantitative growth. It is known that due to the increase in the number of benign diseases of the cervix, the development of precancerous conditions occurs. Benign diseases of the cervix represent the most common gynecological problem, which are often precursors of malignant neoplasms, especially cervical cancer. According to statistics, benign diseases of the cervix cover 25-45% of all gynecological diseases. Among women's oncogynecological diseases, cervical cancer ranks second in the world after breast cancer and ranks first in the mortality rate among oncological diseases in economically underdeveloped countries. We performed a comprehensive clinical and laboratory examination of 130 women aged 18 to 73 with benign cervical diseases. 59 (38.5%) women of reproductive age, as well as 39 (30%) premenopausal and 41 (31.5%) menopausal patients, participated in the study. Detailed anamnesis was collected from all patients, objective and gynecological examination was performed, laboratory and instrumental examinations (USM, IPV DNA, smear microscopy, and PCR bacteriological examination of sexually transmitted infections), simple and extended colposcopy, liquid-based РАР-smear smear and РАР-classic smear examinations were conducted. As a result of the research, the following nosological forms were found in women with benign diseases of the cervix: non-specific vaginitis in 10 (7.7%) cases; ectopia, endocervicitis - 60(46.2%); cervical ectropion - 7(5.4%); cervical polyp - 9(6.9%); cervical leukoplakia - 15(11.5%); atrophic vaginitis - 7(5.4%); condyloma - 12(9.2%); cervical stenosis - 2(1.5%); endometriosis of the cervix - was noted in 8 (6.2%) cases (p<0.001), respectively. Characteristics of the menstrual cycle among the examined women: normal cycle in 97 (74.6%) cases; oligomenorrhea – 23 (17.7%); polymenorrhea – 4(3.1%); algomenorrhea – noted in 6 (4.6%) cases (p<0.001). Cytological examination showed that: the specificity of liquid-based cytology was 76.2%, and the traditional PAP test was set at 70.6%. The overall diagnostic value was calculated to be 86% in liquid-based cytology and 78.5% in conventional PAP tests. Treatment of women with benign diseases of the cervix was carried out by diathermocoagulation method and "FOTEK EA 141M" device. It should be noted that 6 months after the treatment, after treatment with the "FOTEK EA 141M" device, there was no relapse in any patient. Recurrence was found in 23.7% of patients after diathermoelectrocoagulation. Thus, it is clear from the above that the study of cervical pathologies, the determination of optimal examinations, and effective treatment methods is one of the urgent problems facing obstetrics and gynecology.

Keywords: cervical cancer, cytological examination, PAP-smear, non-specific vaginitis

Procedia PDF Downloads 117

Authors: Menna Ewida, Dalal Abou El-Ella, Dina Lasheen, Huessin El-Subbagh

Abstract:

Introduction: Vascular Endothelial Growth Factor receptor (VEGF) is a signal protein produced by cells that stimulates vasculogenesis to create new blood vessels. VEGF family binds to three trans-membrane tyrosine kinase receptors,Dihydrofolate reductase (DHFR) is an enzyme of crucial importance in medicinal chemistry. DHFR catalyzes the reduction 7,8 dihydro-folate to tetrahydrofolate and intimately couples with thymidylate synthase which is a pivotal enzyme that catalysis the reductive methylation of deoxyuridine monophosphate (dUMP) to deoxythymidine monophosphate (dTMP) utilizing N5,N10-methylene tetrahydrofolate as a cofactor which functions as the source of the methyl group. Purpose: Novel substituted Thiazole agents were designed as DHFR and VEGF-TK inhibitors with increased synergistic activity and decreased side effects. Methods: Five series of compounds were designed with a rational that mimic the pharmacophoric features present in the reported active compounds that target DHFR & VEGFR. These molecules were docked against Methotrexate & Sorafenib as controls. An in silico ADMET study was also performed to validate the bioavailability of the newly designed compounds. The in silico molecular docking & ADMET study were also applied to the non-classical antifolates for comparison. The interaction energy comparable to that of MTX for DHFRI and Sorafenib for VEGF-TKI activity were recorded. Results: Compound 5 exhibited the highest interaction energy when docked against Sorafenib, While Compound 9 showed the highest interaction energy when docked against MTX with the perfect binding mode. Comparable results were also obtained for the ADMET study. Most of the compounds showed absorption within (95-99) zone which varies according to the type of substituents. Conclusions: The Substituted Thiazole Analogues could be a suitable template for antitumor drugs that possess enhanced bioavailability and act as DHFR and VEGF-TK inhibitors.

Keywords: anti-tumor agents, DHFR, drug design, molecular modeling, VEGFR-TKIs

Procedia PDF Downloads 235

Authors: Chloe Richards, Adrian Delgado Ollero, Yan Delaure, Fiona Regan

Abstract:

Growing concern about the natural environment has accelerated the search for non-toxic, but at the same time, economically reasonable, antifouling materials. Bioinspired surfaces, due to their nano and micro topographical antifouling capabilities, provide a hopeful approach to the design of novel antifouling surfaces. Biological organisms are known to have highly evolved and complex topographies, demonstrating antifouling potential, i.e. shark skin. Previous studies have examined the antifouling ability of topographic patterns, textures and roughness scales found on natural organisms. One of the mechanisms used to explain the adhesion of cells to a substrate is called attachment point theory. Here, the fouling organism experiences increased attachment where there are multiple attachment points and reduced attachment, where the number of attachment points are decreased. In this study, an attempt to characterize the microtopography of the common brill fish, Scophthalmus rhombus, was undertaken. Scophthalmus rhombus is a small flatfish of the family Scophthalmidae, inhabiting regions from Norway to the Mediterranean and the Black Sea. They reside in shallow sandy and muddy coastal areas at depths of around 70 – 80 meters. Six engineered surfaces (inspired by the Brill fish scale) produced by a 2-photon polymerization (2PP) process were evaluated for their potential as an antifouling solution for incorporation onto tidal energy blades. The micro-textures were analyzed for their AF potential under both static and dynamic laboratory conditions using two laboratory grown diatom species, Amphora coffeaeformis and Nitzschia ovalis. The incorporation of a surface topography was observed to cause a disruption in the growth of A. coffeaeformis and N. ovalis cells on the surface in comparison to control surfaces. This work has demonstrated the importance of understanding cell-surface interaction, in particular, topography for the design of novel antifouling technology. The study concluded that biofouling can be controlled by physical modification, and has contributed significant knowledge to the use of a successful novel bioinspired AF technology, based on Brill, for the first time.

Keywords: attachment point theory, biofouling, Scophthalmus rhombus, topography

Procedia PDF Downloads 107

Authors: Valeria Arcucci, Musarat Ishaq, Steven A. Stacker, Greg J. Goodall, Marc G. Achen

Abstract:

Metastasis is the lethal aspect of cancer for most patients. Remodelling of lymphatic vessels associated with a tumour is a key initial step in metastasis because it facilitates the entry of cancer cells into the lymphatic vasculature and their spread to lymph nodes and distant organs. Although it is clear that vascular endothelial growth factors (VEGFs), such as VEGF-C and VEGF-D, are key drivers of lymphatic remodelling, the means by which many signaling pathways in endothelial cells are coordinately regulated to drive growth and remodelling of lymphatics in cancer is not understood. We seek to understand the broader molecular mechanisms that control cancer metastasis, and are focusing on microRNAs, which coordinately regulate signaling pathways involved in complex biological responses in health and disease. Here, using small RNA sequencing, we found that a specific microRNA, miR-132, is upregulated in expression in lymphatic endothelial cells (LECs) in response to the lymphangiogenic growth factors. Interestingly, ectopic expression of miR-132 in LECs in vitro stimulated proliferation and tube formation of these cells. Moreover, miR-132 is expressed in lymphatic vessels of a subset of human breast tumours which were previously found to express high levels of VEGF-D by immunohistochemical analysis on tumour tissue microarrays. In order to dissect the complexity of regulation by miR-132 in lymphatic biology, we performed Argonaute HITS-CLIP, which led us to identify the miR-132-mRNA interactome in LECs. We found that this microRNA in LECs is involved in the control of many different pathways mainly involved in cell proliferation and regulation of the extracellular matrix and cell-cell junctions. We are now exploring the functional significance of miR-132 targets in the biology of LECs using biochemical techniques, functional in vitro cell assays and in vivo lymphangiogenesis assays. This project will ultimately define the molecular regulation of lymphatic remodelling by miR-132, and thereby identify potential therapeutic targets for drugs designed to restrict the growth and remodelling of tumour lymphatics resulting in metastatic spread.

Keywords: argonaute HITS-CLIP, cancer, lymphatic remodelling, miR-132, VEGF

Procedia PDF Downloads 128

Authors: Eman M. Sarhan, Doaa A. Ghareeb, Gabriella Ortore, Amr A. Amara, Mohamed M. El-Sayed

Abstract:

Drug salting and nanoparticle-based drug delivery formulations are considered to be an effective means for rendering the hydrophobic drugs’ nano-scale dispersion in aqueous media, and thus circumventing the pitfalls of their poor solubility as well as enhancing their membrane permeability. The current study aims to increase the bioavailability of quaternary ammonium berberine through acid salting and biodegradable bovine serum albumin (BSA)-based nanoparticulate drug formulation. Berberine hydroxide (BBR-OH) that was chemically synthesized by alkalization of the commercially available berberine hydrochloride (BBR-HCl) was then acidified to get Di-berberine sulfate (BBR)₂SO₄. The purified crystals were spectrally characterized. The desolvation technique was optimized for the preparation of size-controlled BSA-BBR-HCl, BSA-BBR-OH, and BSA-(BBR)₂SO₄ nanoparticles. Particle size, zeta potential, drug release, encapsulation efficiency, Fourier transform infrared spectroscopy (FTIR), tandem MS-MS spectroscopy, energy-dispersive X-ray spectroscopy (EDX), scanning and transmitting electron microscopic examination (SEM, TEM), in vitro bioactivity, and in silico drug-polymer interaction were determined. BSA (PDB ID; 4OR0) protonation state at different pH values was predicted using Amber12 molecular dynamic simulation. Then blind docking was performed using Lamarkian genetic algorithm (LGA) through AutoDock4.2 software. Results proved the purity and the size-controlled synthesis of berberine-BSA-nanoparticles. The possible binding poses, hydrophobic and hydrophilic interactions of berberine on BSA at different pH values were predicted. Antioxidant, anti-hemolytic, and cell differentiated ability of tested drugs and their nano-formulations were evaluated. Thus, drug salting and the potentially effective albumin berberine nanoparticle formulations can be successfully developed using a well-optimized desolvation technique and exhibiting better in vitro cellular bioavailability.

Keywords: berberine, BSA, BBR-OH, BBR-HCl, BSA-BBR-HCl, BSA-BBR-OH, (BBR)₂SO₄, BSA-(BBR)₂SO₄, FTIR, AutoDock4.2 Software, Lamarkian genetic algorithm, SEM, TEM, EDX

Procedia PDF Downloads 174

Authors: Shayli Varasteh Moradi, Wayne A. Johnston, Dejan Gagoski, Kirill Alexandrov

Abstract:

The demand for a rapid and more efficient technique to identify protein-protein interaction particularly in the areas of therapeutics and diagnostics development is growing. The method described here is a rapid in vitro protein-protein interaction analysis approach based on AlphaLISA technology combined with Leishmania tarentolae cell-free protein production (LTE) system. Cell-free protein synthesis allows the rapid production of recombinant proteins in a multiplexed format. Among available in vitro expression systems, LTE offers several advantages over other eukaryotic cell-free systems. It is based on a fast growing fermentable organism that is inexpensive in cultivation and lysate production. High integrity of proteins produced in this system and the ability to co-express multiple proteins makes it a desirable method for screening protein interactions. Following the translation of protein pairs in LTE system, the physical interaction between proteins of interests is analysed by AlphaLISA assay. The assay is performed using unpurified in vitro translation reaction and therefore can be readily multiplexed. This approach can be used in various research applications such as epitope mapping, antigen-antibody analysis and protein interaction network mapping. The intra-viral protein interaction network of Zika virus was studied using the developed technique. The viral proteins were co-expressed pair-wise in LTE and all possible interactions among viral proteins were tested using AlphaLISA. The assay resulted to the identification of 54 intra-viral protein-protein interactions from which 19 binary interactions were found to be novel. The presented technique provides a powerful tool for rapid analysis of protein-protein interaction with high sensitivity and throughput.

Keywords: AlphaLISA technology, cell-free protein expression, epitope mapping, Leishmania tarentolae, protein-protein interaction

Procedia PDF Downloads 237