Search results for: branched chain amino acid

Authors: Nazish Badar

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In early 2009, Pandemic type A (H1N1) Influenza virus emerged globally. Since then, it has continued circulation causing considerable morbidity and mortality. The purpose of this study was to evaluate the evolutionary changes in Influenza A (H1N1) pdm09 viruses from 2009-15 and their relevance with the current vaccine viruses. Methods: Respiratory specimens were collected with influenza-like illness and Severe Acute Respiratory Illness. Samples were processed according to CDC protocol. Sequencing and phylogenetic analysis of Haemagglutinin (HA) and neuraminidase (NA) genes was carried out comparing representative isolates from Pakistan viruses. Results: Between Jan2009 - Feb 2016, 1870 (13.2%) samples were positive for influenza A out of 14086. During the pandemic period (2009–10), Influenza A/ H1N1pdm 09 was the dominant strain with 366 (45%) of total influenza positives. In the post-pandemic period (2011–2016), a total of 1066 (59.6%) cases were positive Influenza A/ H1N1pdm 09 with co-circulation of different Influenza A subtypes. Overall, the Pakistan A(H1N1) pdm09 viruses grouped in two genetic clades. Influenza A(H1N1)pdm09 viruses only ascribed to Clade 7 during the pandemic period whereas viruses belong to clade 7 (2011) and clade 6B (2015) during the post-pandemic years. Amino acid analysis of the HA gene revealed mutations at positions S220T, I338V and P100S specially associated with outbreaks in all the analyzed strains. Sequence analyses of post-pandemic A(H1N1)pdm09 viruses showed additional substitutions at antigenic sites; S179N,K180Q (SA), D185N, D239G (CA), S202A (SB) and at receptor binding sites; A13T, S200P when compared with pandemic period. Substitution at Genetic markers; A273T (69%), S200P/T (15%) and D239G (7.6%) associated with severity and E391K (69%) associated with virulence was identified in viruses isolated during 2015. Analysis of NA gene revealed outbreak markers; V106I (23%) among pandemic and N248D (100%) during post-pandemic Pakistan viruses. Additional N-Glycosylation site; HA S179N (23%), NA I23T(7.6%) and N44S (77%) in place of N386K(77%) were only found in post-pandemic viruses. All isolates showed histidine (H) at position 275 in NA indicating sensitivity to neuraminidase inhibitors. Conclusion: This study shows that the Influenza A(H1N1)pdm09 viruses from Pakistan clustered into two genetic clades, with co-circulation of some variants. Certain key substitutions in the receptor binding site and few changes indicative of virulence were also detected in post-pandemic strains. Therefore, it is imperative to continue monitoring of the viruses for early identification of potential variants of high virulence or emergence of drug-resistant variants.

Keywords: Influenza A (H1N1) pdm09, evolutionary analysis, post pandemic period, Pakistan

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Authors: Jianuo Sun, Jinghan Wang, Sirui Zhang, Chenhan Xu, Hongxia Hao, Hong Zhou

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In recent years, the diversification and industrialization of drug-related crimes have posed significant threats to public health and safety globally. The widespread and increasingly younger demographics of drug users and the persistence of drug-impaired driving incidents underscore the urgency of this issue. Drug detection, a specialized forensic activity, is pivotal in identifying and analyzing substances involved in drug crimes. It relies on pharmacological and chemical knowledge and employs analytical chemistry and modern detection techniques. However, current drug detection methods are limited by their inability to perform semi-quantitative, real-time field analyses. They require extensive, complex laboratory-based preprocessing, expensive equipment, and specialized personnel and are hindered by long processing times. This study introduces an alternative approach using nucleic acid aptamers and Aggregation-Induced Emission (AIE) technology. Nucleic acid aptamers, selected artificially for their specific binding to target molecules and stable spatial structures, represent a new generation of biosensors following antibodies. Rapid advancements in AIE technology, particularly in tetraphenyl ethene-based luminous, offer simplicity in synthesis and versatility in modifications, making them ideal for fluorescence analysis. This work successfully synthesized, isolated, and purified an AIE molecule and constructed a probe comprising the AIE molecule, nucleic acid aptamers, and exonuclease for cocaine detection. The probe demonstrated significant relative fluorescence intensity changes and selectivity towards cocaine over other drugs. Using 4-Butoxytriethylammonium Bromide Tetraphenylethene (TPE-TTA) as the fluorescent probe, the aptamer as the recognition unit, and Exo I as an auxiliary, the system achieved rapid detection of cocaine within 5 mins in aqueous and urine, with detection limits of 1.0 and 5.0 µmol/L respectively. The probe-maintained stability and interference resistance in urine, enabling quantitative cocaine detection within a certain concentration range. This fluorescent sensor significantly reduces sample preprocessing time, offers a basis for rapid onsite cocaine detection, and promises potential for miniaturized testing setups.

Keywords: drug detection, aggregation-induced emission (AIE), nucleic acid aptamer, exonuclease, cocaine

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Authors: Amina Adala, Nadra Debbache, Tahar Sehili

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Coordination polymers and uniformly {[Zn(II)(BIPY)(Pht)]n} (1), {[Zn (HYD)(Pht)]n} (2) (BIPY = 4,4’ bipyridine, Pht = terephtalic acid, HYD = 8-hydroxyquinoline) have been successfully synthesized by a hydrothermal process using aqueous zinc solution. The as-prepared compounds phases were characterized by X-ray diffraction (XRD), Fourier Transform Infrared spectroscopy, UV-visible spectroscopy, thermogravimetric analysis (TGA), and the electrochemistry study by the voltammetry cyclic. The results showed a crystalline phase for CP1 however, CP2 requires recrystallization; the FTIR showed the presence of characteristic bands of all ligands; besides that, TGA shows thermal stability up to 300°C. The electrochemistry study showed a good charge transfer between the ligands and Zn metal for the two components. UV-Vis measurement showed strong absorption in a wide range from UV to visible light with a band gap of 2.69 eV for CP1 and 2.56 eV for CP2, smaller than that of ZnO. This represents an alternative to using ZnO. The Ibuprofen IBP decomposition kinetics of 5.10⁻⁵ mol.L⁻¹ under solar and artificial light were studied for different irradiation conditions. Good photocatalytic properties were observed due to their high surface area.

Keywords: metal-organic frameworks, photocatalysis, photodegradation, organic pollutant, ibuprofen

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Authors: Ässia Boukhatmi, Roger Nyffenegger

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The expansion of solar energy as a means to address the climate crisis is undisputed, but the increasing number of new photovoltaic (PV) modules being put on the market is simultaneously leading to increased challenges in terms of managing the growing waste stream. Many of the discarded modules are still fully functional but are often damaged by improper handling after disassembly or not properly tested to be considered for a second life. In addition, the collection rate for dismantled PV modules in several European countries is only a fraction of previous projections, partly due to the increased number of illegal exports. The underlying problem for those market imperfections is an insufficient data exchange between the different actors along the PV value chain, as well as the limited traceability of PV panels during their lifetime. As part of the Horizon 2020 project CIRCUSOL, an asset database prototype was developed to tackle the described problems. In an iterative process applying the design science research methodology, different business models, as well as the technical implementation of the database, were established and evaluated. To explore the requirements of different stakeholders for the development of the database, surveys and in-depth interviews were conducted with various representatives of the solar industry. The proposed database prototype maps the entire value chain of PV modules, beginning with the digital product passport, which provides information about materials and components contained in every module. Product-related information can then be expanded with performance data of existing installations. This information forms the basis for the application of data analysis methods to forecast the appropriate end-of-life strategy, as well as the circular economy potential of PV modules, already before they arrive at the recycling facility. The database prototype could already be enriched with data from different data sources along the value chain. From a business model perspective, the database offers opportunities both in the area of reuse as well as with regard to the certification of sustainable modules. Here, participating actors have the opportunity to differentiate their business and exploit new revenue streams. Future research can apply this approach to further industry and product sectors, validate the database prototype in a practical context, and can serve as a basis for standardization efforts to strengthen the circular economy.

Keywords: business model, circular economy, database, design science research, solar industry

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Authors: Emad Al-Abdallat, Faris G. Bakri, Azmi Mahafza, Rayyan Al Ali, Nidaa Ababneh, Ahmed Idhair

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Background: Morgues are high-risk areas for the spread of infection from the cadavers to the staff during the postmortem examination. Infection can spread from corpses to workers by the airborne route, by direct contact, or from needle and sharp object injuries. Objective: Knowledge about the prevalence of these infections among autopsies is prudent to appreciate any risk of transmission and to further enforce safety measures. Method: A total of 242 autopsies were tested. Age ranged from 3 days to 94 years (median 75.5 years, mean 45.3 (21.9 ± SD)). There were 172 (71%) males. Results: The cause of death was considered natural in 137 (56.6%) cases, accidental in 89 (36.8%), homicidal in 9 (3.7%), suicidal in 4 (1.7%), and unknown in 3 (1.2%). Hepatitis B surface antigen was positive in 5 (2.1%) cases. Hepatitis C virus antibody was detected in 5 (2.1%) cases and the hepatitis C virus polymerase chain reaction was positive in 2 of them (0.8%). HIV antibody was not detected in any of the cases. Conclusions: Autopsies can be associated with exposure to blood borne viruses. Autopsies performed during the study period were tested for hepatitis B surface antigen, hepatitis C virus antibody, and human immunodeficiency virus antibody. Positive tests were subsequently confirmed by polymerase chain reaction. There is low prevalence of infections with these viruses in our autopsy cases. However, the risk of transmission remains a threat. Healthcare workers in the forensic departments should adhere to standard precautions.

Keywords: autopsy, hepatitis B virus, hepatitis C virus, human immunodeficiency virus, Jordan

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Authors: Boris M. Popović, Tatjana Jurić, Bojana Blagojević, Denis Uka, Ružica Ždero Pavlović

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Green solvents are environmentally friendly and greatly improve the sustainability of chemical processes. There is a growing interest in the green extraction of polyphenols from fruits. In this study, we consider three Natural Deep Eutectic Solvents (NADES) systems based on choline chloride as a hydrogen bond acceptor and malic acid, urea, and fructose as hydrogen bond donors. NADES systems were prepared by heating and stirring, ultrasound, and microwave (MW) methods. Sour cherry pomace was used as a natural source of polyphenols. Polyphenol extraction from cherry pomace was performed by ultrasound-assisted extraction and microwave-assisted extraction and compared with conventional heat and stirring method extraction. It was found that MW-assisted preparation of NADES was the fastest, requiring less than 30 s. Also, MW extraction of polyphenols was the most rapid, with less than 5 min necessary for the extract preparation. All three NADES systems were highly efficient for anthocyanin extraction, but the most efficient was the system with malic acid as a hydrogen bond donor (yield of anthocyanin content was enhanced by 62.33% after MW extraction with NADES compared with the conventional solvent).

Keywords: anthocyanins, green extraction, NADES, polyphenols

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Authors: Abhishekkumar Ramasamy, Parameshwari

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Chitosan is the deacyelitated derivative of chitin, the second most abundant biopolymer just after cellulose. Without doubt, its biomedical usages have gained more importance among the vast variety of chitosan applications owing to its good biocompatibility and biodegradability. In recent years, particular interest has been devoted to chitosan hydrogels as a promising alternative in competition with conventional sutures or bioadhesives. Different parameters such as acid type and concentration, and degree of deacetylation (DD%) of chitosan, were altered to modify hydrogel properties including viscosity, pH, cohesive strength, and tissue bioadhesiveness. In the current work, we have investigated the effectiveness of chitosan hydrogel encapsulated with tanexamic acid to stop bleeding. Chitosan film was obtained with solubilization of chitosan powder in aqueous acidic media. In vivo experiments have been conducted on rat and rabbit models that provide a convenient way to evaluate the efficacy of prepared samples. The arteries vein was punctured on the hind limb of the rat and the gauze was been applied on the punchered area. Bioadhesive strength as well as irritant effects were discussed. Samples with higher degree of deacetylation, including Chs-16 and Chs-19 that were dissolved in lactic media showed best sealing effect.

Keywords: chitosan, biocomaptibility, biodegradability, bioadhersive, deacetylation

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Authors: Naruphorn Dararatana, Farzad Seidi, Daniel Crespy

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Protection of metals is a critical issue in industry. The annual cost of corrosion in the world is estimated to be about 2.5 trillion dollars and continuously increases. Therefore, there is a need for developing novel protection approaches to improve corrosion protection. We designed and synthesized smart polymer/corrosion inhibitor conjugates as new generations of corrosion protecting materials. Firstly, a polymerizable acrylate derivative of 8-hydroxyquinoline (8HQ), an effective corrosion inhibitor, containing acid-labile β-thiopropionate linkage was prepared in three steps. Then, it was copolymerized with ethyl acrylate in the presence of 1,1′-azobis(cyclohexanecarbonitrile) (ABCN) by radical polymerization. Nanoparticles with an average diameter of 140 nm were prepared from the polymer conjugate by the miniemulsion-solvent evaporation process. The release behavior of 8HQ from the the nanoparticles was studied in acidic (pH 3.5) and neutral media (pH 7.0). The release profile showed a faster release of 8HQ in acidic medium in comparison with neutral medium. Indeed 100% of 8HQ was released after 14 days in acidic medium whereas only around 15% of 8HQ was released during the same period at neutral pH. Therefore, the polymer conjugate nanoparticles are suitable materials as additives or to form coatings on metal substrates for corrosion protection.

Keywords: Corrosion inhibitor, 8-Hydroxyquinoline, Polymer conjugated, β-Thiopropionate

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Authors: S. Ibrahim, U. B. Abubakar, S. Danbirni, A. Usman, F. M. Ballah, C. A. Kudi, L. Lawson, G. H. Abdulrazak, I. A. Abdulkadir

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Performing culture and characterization of mycobacteria in low resource settings like Nigeria is a very difficult task to undertake because of the very few and limited laboratories carrying out such an experiment; this is a largely due to stringent and laborious nature of the tests. Hence, a rapid, simple and accurate test for characterization is needed. The “SD BIOLINE TB Ag MPT 64 Rapid ®” is a simple and rapid immunochromatographic test used in differentiating Mycobacteria into Mycobacterium tuberculosis (NTM). The 100 sputa were obtained from patients suspected to be infected with tuberculosis and presented themselves to hospitals for check-up and treatment were involved in the study. The samples were cultured in a class III Biosafety cabinet and level III biosafety practices were followed. Forty isolates were obtained from the cultured sputa, and there were identified as Acid-fast bacilli (AFB) using Zeihl-Neelsen acid-fast stain. All the isolates (AFB positive) were then subjected to the SD BIOLINE Analyses. A total of 31 (77.5%) were characterized as MTBC, while nine (22.5%) were NTM. The total turnaround time for the rapid assay was just 30 minutes as compared to a few days of phenotypic and genotypic method. It was simple, rapid and reliable test to differentiate MTBC from NTM.

Keywords: culture, mycobacteria, non tuberculous mycobacterium, SD Bioline

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Authors: Al-Braa Kashegari, Ali M. El-Halawany, Akram A. Shalabi, Sabrin R. M. Ibrahim, Hossam M. Abdallah

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Chemical investigation of Chrozophora oblongifolia resulted in the isolation of five major compounds that were identified as apeginin-7-O-glucoside (1), quercetin-3-O-glucuronic acid (2), quercetin-3-O-glacturonic acid (3), rutin (4), and 1,3,6-trigalloyl glucose (5). The identity of isolated compounds was assessed by different spectroscopic methods, including one- and two-dimensional NMR. The isolated compounds were tested for their antioxidant activity using different assays viz., DPPH, FRAP, ABTS, ORAC, and metal chelation effects. In addition, the inhibition of target enzymes involved in the metabolic syndrome, such as alpha-glucosidase and pancreatic lipase, were carried out. Moreover, the effect of the compounds on the advanced glycation end-products (AGEs) as one of the major complications of oxidative stress and hyperglycemia in metabolic syndromes were carried out using BSA‐fructose (bovine serum albumin), BSA-methylglyoxal, and arginine methylglyoxal models. The pure isolates showed a protective effect in metabolic syndromes as well as promising antioxidant activity. The results showed potent activity of compound 5 in all measured parameters meanwhile, none of the tested compounds showed activity against pancreatic lipase.

Keywords: Chrozophora oblongifolia, antioxidant, pancreatic lipase, metabolic syndromes

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Authors: El-Said I. El-Shafey, Syeda Naheed F. Ali, Saleh S. Al-Busafi, Haider A. J. Al-Lawati

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Date palm leaflets were utilized as a precursor for activated carbon (AC) preparation using KOH activation. AC produced was oxidized using nitric acid producing oxidized activated carbon (OAC). OAC that possesses acidic surface was surface functionalized to produce basic activated carbons using linear diamine compounds (ethylene diamine and propylene diamine). OAC was also functionalized to produce hydrophobic activated carbons using ethylamine (EA) and aniline (AN). Dehydrated carbon was also prepared from date palm leaflets using sulfuric acid dehydration/ oxidation and was surface functionalized in the same way as AC. Nitric acid oxidation was not necessary for DC as it is acidic carbon. The surface area of AC is high (823 m2/g) with microporosity domination, however, after oxidation and surface functionalization, both the surface area and surface microporosity decrease tremendously. DC surface area was low (15 m2/g) with mesoporosity domination. Surface functionalization has decreased the surface area of activated carbons. FTIR spectra show that -COOH group on DC and OAC almost disappeared after surface functionalization. The surface chemistry of all carbons produced was tested for pHzpc, basic sites, boehm titration, thermogravimetric analysis and zeta potential measurement. Scanning electron microscopy and energy dispersive spectroscopy in addition to CHN elemental analysis were also carried out. DC and OAC possess low pHzpc and high surface functionality, however, basic and hydrophobic carbons possess high pHzpc and low surface functionality. The different behavior of carbons is related to their different surface chemistry. Methylene blue adsorption was found to be faster on hydrophobic carbons based on AC and DC. The Larger adsorption capacity of methylene blue was found for hydrophobic carbons. Dominating adsorption forces of methylene blue varies from carbon to another depending on its surface nature. Sorption forces include hydrophobic forces, H-bonding, electrostatic interactions and van der Waals forces.

Keywords: carbon, acidic, basic, hydrophobic

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Authors: Attila Kiss, Erzsébet Némedi, Zoltán Naár

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As natural prebiotic (non-digestible) carbohydrates stimulate the growth of colon microflora and contribute to maintain the health of the host, analytical studies aiming at revealing the chemical behavior of these beneficial food components came to the forefront of interest. Food processing (especially baking) may lead to a significant conversion of the parent compounds, hence it is of utmost importance to characterize the transformation patterns and the plausible decomposition products formed by thermal degradation. The relevance of this work is confirmed by the wide-spread use of these carbohydrates (fructo-oligosaccharides, cyclodextrins, raffinose and resistant starch) in the food industry. More and more functional foodstuffs are being developed based on prebiotics as bioactive components. 12 different types of oligosaccharides have been investigated in order to reveal their thermal degradation characteristics. Different carbohydrate derivatives (D-fructose and D-glucose oligomers and polymers) have been exposed to elevated temperatures (150 °C 170 °C, 190 °C, 210 °C, and 220 °C) for 10 min. An advanced HPLC method was developed and used to identify the decomposition products of carbohydrates formed as a consequence of thermal treatment. Gradient elution was applied with binary solvent elution (acetonitrile, water) through amine based carbohydrate column. Evaporative light scattering (ELS) proved to be suitable for the reliable detection of the UV/VIS inactive carbohydrate degradation products. These experimental conditions and applied advanced techniques made it possible to survey all the formed intermediers. Change in oligomer distribution was established in cases of all studied prebiotics throughout the thermal treatments. The obtained results indicate increased extent of chain degradation of the carbohydrate moiety at elevated temperatures. Prevalence of oligomers with shorter chain length and even the formation of monomer sugars (D-glucose and D-fructose) might be observed at higher temperatures. Unique oligomer distributions, which have not been described previously are revealed in the case of each studied, specific carbohydrate, which might result in various prebiotic activities. Resistant starches exhibited high stability when being thermal treated. The degradation process has been modeled by a plausible reaction mechanism, in which proton catalyzed degradation and chain cleavage take place.

Keywords: prebiotics, thermal degradation, fructo-oligosaccharide, HPLC, ELS detection

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Authors: Van-Thanh Ho, Jaiyoung Ryu

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In soft robotics, the optimization of fluid dynamics through pneumatic methods plays a pivotal role in enhancing operational efficiency and reducing energy loss. This is particularly crucial when replacing conventional techniques such as cable-driven electromechanical systems. The pneumatic model employed in this study represents a sophisticated framework designed to efficiently channel pressure from a high-pressure reservoir to various muscle locations on the robot's body. This intricate network involves a branching system of tubes. The study introduces a comprehensive pneumatic model, encompassing the components of a reservoir, tubes, and Pneumatically Actuated Muscles (PAM). The development of this model is rooted in the principles of shock tube theory. Notably, the study leverages experimental data to enhance the understanding of the interplay between the PAM structure and the surrounding fluid. This improved interactive approach involves the use of morphing motion, guided by a contraction function. The study's findings demonstrate a high degree of accuracy in predicting pressure distribution within the PAM. The model's predictive capabilities ensure that the error in comparison to experimental data remains below a threshold of 10%. Additionally, the research employs a machine learning model, specifically a surrogate model based on the Kriging method, to assess and quantify uncertainty factors related to the initial reservoir pressure and tube diameter. This comprehensive approach enhances our understanding of pneumatic soft robotics and its potential for improved operational efficiency.

Keywords: pneumatic artificial muscles, pressure drop, morhing motion, branched network, surrogate model

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Authors: Makhdoom Saad Wasim Ghouri, Muneeba Butt, Mohammad Ashraf Tahir, Rashid Bhatti, Akbar Ali, Abdul Rehman, Abdul Basit, Muzzamel Rehman, Shahbaz Aslam, Farakh Mansoor, Ahmad Fayyaz, Hadia Siddiqui

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Lip print analysis (Cheiloscopy) is the new emerging technique that might be the guardian angel in establishing the personal identity. Cheiloscopy is basically the study of elevations and depressions present on the external surface of the lips. In our study, 600 lip prints samples were taken (300 males and 300 females). Lip prints of each individual were divided into four quadrants and the upper middle portion. For general classification, middle part of the lower lip almost 10 mm wide would be taken into consideration. After analysis of lip-prints, our results show that lip prints are the unique and permanent character of every individual. No two lip print was matched with each other even of the identical twins. Our study reveals that there is equal distribution of lip print patterns among all the four quadrants of lips and the upper middle portion; these distributions were statistically analyzed by applying chi-square test which shows the significant results. In general classification, 5 lip print types/patterns were studied, Type 1 (Vertical lines), Type 2 (Branched pattern), Type 3 (Intersected pattern), Type 4 (Reticular pattern) and Type 5 (Undetermined). Type 1 and Type 2 were found to be the most frequent patterns in female population, while Type 3 and Type 4 most commonly found in male population. These results were also analyzed by applying Chi-square test, and the results show significance statistically. Thus, establishing sex determination on the basis of lip print types among the gender. Type 5 was the least common pattern among genders.

Keywords: cheiloscopy, distribution, quadrants, sex determination

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Authors: Baker Ssekitto, Ambrose Mbogo

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The study reveals that indeed agriculture employs over 70% of Uganda’s population, of which majority are youth and women. The study further reveals that over 70% of the farmers are smallholder farmers based in rural areas, whose operations are greatly affected by; climate change, weak digital skills, limited access to productivity knowledge along value chains, limited access to quality farm inputs, weak logistics systems, limited access to quality extension services, weak business intelligence, limited access to quality markets among others. It finds that the emerging 4th industrial revolution powered by artificial intelligence, 5G and data science will provide possibilities of addressing some of these challenges. Furthermore, the study finds that despite rapid development of ICT4Agric Innovation, their uptake is constrained by a number of factors including; limited awareness of these innovations, low internet and smart phone penetration especially in rural areas, lack of appropriate digital skills, inappropriate programmes implementation models which are project and donor driven, limited articulation of value addition to various stakeholders among others. Majority of farmers and other value chain actors lacked knowledge and skills to harness the power of ICTs, especially their application of ICTs in monitoring and evaluation on quality of service in the extension system and farm level processes.

Keywords: artificial intelligence, productivity, ICT4agriculture, value chain, logistics

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Authors: Soma Banerjee, Aamen Talukdar, Argha Mandal, Dipankar Chaudhuri

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Japanese Encephalitis is a major infectious disease with nearly half the world’s population living in areas where it is prevalent. Currently, treatment for it involves only supportive care and symptom management through vaccination. Due to the lack of antiviral drugs against Japanese Encephalitis Virus (JEV), the quest for such agents remains a priority. For these reasons, simulation studies of drug targets against JEV are important. Towards this purpose, docking experiments of the kinase inhibitors were done against the chosen target NS3 helicase as it is a nucleoside binding protein. Previous efforts regarding computational drug design against JEV revealed some lead molecules by virtual screening using public domain software. To be more specific and accurate regarding finding leads, in this study a proprietary software Schrödinger-GLIDE has been used. Druggability of the pockets in the NS3 helicase crystal structure was first calculated by SITEMAP. Then the sites were screened according to compatibility with ATP. The site which is most compatible with ATP was selected as target. Virtual screening was performed by acquiring ligands from databases: KinaseSARfari, KinaseKnowledgebase and Published inhibitor Set using GLIDE. The 25 ligands with best docking scores from each database were re-docked in XP mode. Protein structure alignment of NS3 was performed using VAST against MMDB, and similar human proteins were docked to all the best scoring ligands. The low scoring ligands were chosen for further studies and the high scoring ligands were screened. Seventy-three ligands were listed as the best scoring ones after performing HTVS. Protein structure alignment of NS3 revealed 3 human proteins with RMSD values lesser than 2Å. Docking results with these three proteins revealed the inhibitors that can interfere and inhibit human proteins. Those inhibitors were screened. Among the ones left, those with docking scores worse than a threshold value were also removed to get the final hits. Analysis of the docked complexes through 2D interaction diagrams revealed the amino acid residues that are essential for ligand binding within the active site. Interaction analysis will help to find a strongly interacting scaffold among the hits. This experiment yielded 21 hits with the best docking scores which could be investigated further for their drug like properties. Aside from getting suitable leads, specific NS3 helicase-inhibitor interactions were identified. Selection of Target modification strategies complementing docking methodologies which can result in choosing better lead compounds are in progress. Those enhanced leads can lead to better in vitro testing.

Keywords: antivirals, docking, glide, high-throughput virtual screening, Japanese encephalitis, ns3 helicase

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Authors: Anis A. Ansari, M. Kamil

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The development of 3D printing technology has allowed the manufacturing industry to create parts with a high degree of automation, increased design freedom, and improved mechanical performance. Fused deposition modelling (FDM) is a 3D printing technique in which successive layers of thermoplastic polymer are deposited and controlled to create a three-dimensional product. In this study, process parameters such as nozzle temperature and printing speed were chosen to investigate their effects on hardness properties. 3D printed specimens were fabricated by an FDM 3D printer from Polylactic acid (PLA) polymer. After analysis, it was observed that the hardness property is much influenced by print speed and nozzle temperature parameters. Maximum hardness was achieved at higher print speed which indicates that the Shore D hardness is directly proportional to the print speed. Moreover, at higher print speed, it has no significant dependence on the nozzle temperature. Hardness is also influenced by nozzle temperature, though to a lesser extent. The hardness slightly lowers when the nozzle temperature is raised from 190 to 210 oC, but due to improved bonding between each raster, a further rise in temperature increases the hardness property.

Keywords: 3D printing, fused deposition modeling (FDM), polylactic acid (PLA), print speed, nozzle temperature, hardness property

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Authors: Wei Xiao, Gangzhi Cai, Xingliang Qin, Hongyan Ren, Zaidong Hua, Zhe Zhu, Hongwei Xiao, Ximin Zheng, Jie Yao, Yanzhen Bi

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Myostatin (MSTN) is the master regulator of double muscling phenotype with overgrown muscle and decreased fatness in animals, but its action mode to regulate fat deposition remains to be elucidated. In this study a swin-specific pathway through which MSTN acts to regulate the fat deposition was deciphered. Deep sequenincing of the mRNA and miRNA of fat tissues of MSTN knockout (KO) and wildtype (WT) pigs discovered the positive correlation of myocyte enhancer factor 2C (MEF2C) and fat-inhibiting miR222 expression, and the inverse correlation of miR222 and stearoyl-CoA desaturase 5 (SCD5) expression. SCD5 is rodent-absent and expressed only in pig, sheep and cattle. Fatty acid spectrum of fat tissues revealed a lower percentage of oleoyl-CoA (18:1) and palmitoleyl CoA (16:1) in MSTN KO pigs, which are the catalyzing products of SCD5-mediated desaturation of steroyl CoA (18:0) and palmitoyl CoA (16:0). Blood metrics demonstrated a 45% decline of triglyceride (TG) content in MSTN KO pigs. In light of these observations we hypothesized that MSTN might act through MEF2C/miR222/SCD5 pathway to regulate desaturation of fatty acid as well as triglyceride synthesis in pigs. To this end, real-time PCR and Western blotting were carried out to detect the expression of the three genes stated above. These experiments showed that MEF2C expression was up-regulated by nearly 2-fold, miR222 up-regulated by nearly 3-fold and SCD5 down-regulated by nearly 50% in MSTN KO pigs. These data were consistent with the expression change in deep sequencing analysis. Dual luciferase reporter was then used to confirm the regulation of MEF2C upon the promoter of miR222. Ecotopic expression of MEF2C in preadipocyte cells enhanced miR222 expression by 3.48-fold. CHIP-PCR identified a putative binding site of MEF2C on -2077 to -2066 region of miR222 promoter. Electrophoretic mobility shift assay (EMSA) demonstrated the interaction of MEF2C and miR222 promoter in vitro. These data indicated that MEF2C transcriptionally regulates the expression of miR222. Next, the regulation of miR222 on SCD5 mRNA as well as its physiological consequences were examined. Dual luciferase reporter testing revealed the translational inhibition of miR222 upon the 3´ UTR (untranslated region) of SCD5 in preadipocyte cells. Transfection of miR222 mimics and inhibitors resulted in the down-regulation and up-regulation of SCD5 in preadipocyte cells respectively, consistent with the results from reporter testing. RNA interference of SCD5 in preadipocyte cells caused 26.2% reduction of TG, in agreement with the results of TG content in MSTN KO pigs. In summary, the results above supported the existence of a molecular pathway that MSTN signals through MEF2C/miR222/SCD5 to regulate the fat deposition in pigs. This swine-specific pathway offers potential molecular markers for the development and breeding of a new pig line with optimised fatty acid composition. This would benefit human health by decreasing the takeup of saturated fatty acid.

Keywords: fat deposition, MEF2C, miR222, myostatin, SCD5, pig

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Authors: Ramachandra Pradhan, Sandeep Singh Rama, Sabyasachi Mishra

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Jackfruit (Artocarpus heterophyllus L.) in its tender form is consumed as a vegetable and popular for its flavour, colour and meat like texture. In South Asian countries like Bangladesh, India, Pakistan and Indonesia the market value for tender jackfruit is very high. However, due to lack of technology the marketing and transportation of the fruit is a challenge. The processing activities like washing, sorting, peeling and cutting enhances oxidative stress in fresh cut jackfruit. It is also having the ill effects on quality of fresh cut tender jackfruit by an increase in microbial contaminations, excessive tissue softening, and depletion of phytochemicals and browning. Hence, this study was conducted as a solution to the above problem. Fresh cut tender Jackfruit slices were processed by using the independent parameters such as concentration of CaCl2 (2-5%), concentration of citric acid (1-2.5%) and treatment time (4-10 min.) and the depended variables were Browning index (BI), colour change (ΔE), Firmness (F) and Overall all acceptability (OAA) after the treatment. From the response variables the best combination of independent variables was resulted as 3% concentration of CaCl2 and 2% concentration of citric acid for 6 minutes. At these optimised processing treatments, the browning can be prevented for fresh cut tender jackfruit. This technology can be used by the researcher, scientists, industries, etc. for further processing of tender jackfruit.

Keywords: tender jackfruit, browning index, firmness, texture

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Authors: Mesfin Angaw Tesfay

Abstract:

Uridine-5’-diphosphate (UDP)-glucose is one of the most versatile building blocks within the metabolism of prokaryotes and eukaryotes, serving as an activated sugar donor during the glycosylation of natural products. It is formed by the enzyme UDP-glucose pyrophosphorylase (UGPase) using uridine-5′-triphosphate (UTP) and α-d-glucose 1-phosphate as a substrate. Herein, two UGPase genes from Lactobacillus kefiranofaciens ZW3 (LkUGPase) and Rhodococcus wratislaviensis IFP 2016 (RwUGPase) were identified through genome mining approaches. The LkUGPase and RwUGPase have 299 and 306 amino acids, respectively. Both UGPase has the conserved UTP binding site (G-X-G-T-R-X-L-P) and the glucose -1-phosphate binding site (V-E-K-P). The LkUGPase and RwUGPase were cloned in E. coli, and SDS-PAGE analysis showed the expression of both enzymes forming about 36 KDa of protein band after induction. LkUGPase and RwUGPase have an activity of 1549.95 and 671.53 U/mg, respectively. Currently, their kinetic properties are under investigation.

Keywords: UGPase, LkUGPase, RwUGPase, UDP-glucose, glycosylation

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Authors: Pratap K. Chhotaray, Jitendriya Swain, Ashok Mishra, Ramesh L. Gardas

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Ionic liquids (ILs) are molten salts, consist predominantly of ions and found to be liquid below 100°C. The unparalleled growing interest in ILs is based upon their never ending design flexibility. The use of ILs as a co-solvent in binary as well as a ternary mixture with molecular solvents multifold it’s utility. Since polarity is one of the most widely applied solvent concepts which represents simple and straightforward means for characterizing and ranking the solvent media, its study for a binary mixture of ILs is crucial for its widespread application and development. The primary approach to the assessment of solution phase intermolecular interactions, which generally occurs on the picosecond to nanosecond time scales, is to exploit the optical response of photophysical probe. Pyrene butyric acid (PBA) is used as fluorescence probe due to its high quantum yield, longer lifetime and high solvent polarity dependence of fluorescence spectra. Propylammonium formate (PAF) is the IL used for this study. Both the UV-absorbance spectra and steady state fluorescence intensity study of PBA in different concentration of aqueous PAF, reveals that with an increase in PAF concentration, both the absorbance and fluorescence intensity increases which indicate the progressive solubilisation of PBA. Whereas, near about 50% of IL concentration, all of the PBA molecules get solubilised as there are no changes in the absorbance and fluorescence intensity. Furthermore, the ratio II/IV, where the band II corresponds to the transition from S1 (ν = 0) to S0 (ν = 0), and the band IV corresponds to transition from S1 (ν = 0) to S0 (ν = 2) of PBA, indicates that the addition of water into PAF increases the polarity of the medium. Time domain lifetime study shows an increase in lifetime of PBA towards the higher concentration of PAF. It can be attributed to the decrease in non-radiative rate constant at higher PAF concentration as the viscosity is higher. The monoexponential decay suggests that homogeneity of solvation environment whereas the uneven width at full width at half maximum (FWHM) indicates there might exist some heterogeneity around the fluorophores even in the water-IL mixed solvents.

Keywords: fluorescence, ionic liquid, lifetime, polarity, pyrene butyric acid

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Authors: Hye Rim Kang, So Yae Koh, Ji-Yeon Ryu, Chang Kyu Lee, Ji Hee Lim, Hyeon A. Kim, Geun Hyung Im, Somi Kim Cho

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In this study, antioxidant properties and compositional analysis of fermented chocolate were examined. Chocolate was fermented with Lactobacillus plantarum CK10. As fermentation time went by, pH was decreased (5.26±0.02 to 3.98±0.06) while titratable acidity was increased (5.36±0.19 to 13.31±0.34). The total polyphenol contents were maintained through the fermentation. The contents of total polyphenol were slightly increased at 8 hr (6.34±0.12 mg GAE (Gallic acid equivalent)/g), and it reached to comparable levels of the control at 24 hr (control, 5.47±0.36 mg GAE/g); 24 hr, 5.19±0.23 mg GAE/g). Similarly, the total flavonoid contents were not significantly changed during fermentation. The pronounced radical scavenging activities of chocolate, against DPPH-, ABTS-, and Alkyl radical, were observed. The levels of antioxidant activities were not dramatically altered in the course of fermentation. By gas chromatography-mass spectrometry analysis, the increase in lactic acid was measured and four major compounds, HMF, xanthosine, caffeine, and theobromine, were identified. The relative peak area of caffeine and theobromine was considerably changed during fermentation. However, no significant difference in the levels of caffeine and theobromine were observed by high-performance liquid chromatography analysis.

Keywords: antioxidant, chocolate, compositional analysis, fermentation, Lactobaillus plantarum

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Authors: Omar Boutkhoum, Mohamed Hanine, Abdessadek Bendarag

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Sustainable economic growth is nowadays driving firms to extend toward the adoption of many green supply chain management (GSCM) solutions. However, the evaluation and selection of these solutions is a matter of concern that needs very serious decisions, involving complexity owing to the presence of various associated factors. To resolve this problem, a comparative analysis approach based on multi-criteria decision-making methods is proposed for adequate evaluation of sustainable supply chain management solutions. In the present paper, we propose an integrated decision-making model based on FAHP (Fuzzy Analytic Hierarchy Process), TOPSIS (Technique for Order of Preference by Similarity to Ideal Solution) and PROMETHEE (Preference Ranking Organisation METHod for Enrichment Evaluations) to contribute to a better understanding and development of new sustainable strategies for industrial organizations. Due to the varied importance of the selected criteria, FAHP is used to identify the evaluation criteria and assign the importance weights for each criterion, while TOPSIS and PROMETHEE methods employ these weighted criteria as inputs to evaluate and rank the alternatives. The main objective is to provide a comparative analysis based on TOPSIS and PROMETHEE processes to help make sound and reasoned decisions related to the selection problem of GSCM solution.

Keywords: GSCM solutions, multi-criteria analysis, decision support system, TOPSIS, FAHP, PROMETHEE

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Authors: Muhammad SAFDAR, Mehmet Ozaslan, Musarrat Abbas Khan

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Babesiosis is a haemoparasitic disease due to the multiplication of protozoan’s parasite, Babesia ovis in the red blood cells of the host, and contributes numerous economical losses, including sheep and goat ruminants. The early identification and successful treatment of Babesia Ovis spp. belong to the key steps of control and health management of livestock resources. The objective of this study was to construct a polymerase chain reaction (PCR) based method for the detection of Babesia spp. in small ruminants and to determine the risk factors involved in the spreading of babesiosis infections. A total of 100 blood samples were collected from 50 sheep and 50 goats along with different areas of Muzaffargarh, Pakistan, from randomly selected herds. Data on the characteristics of sheep and goats were collected through questionnaires. Of 100 blood samples examined, 18 were positive for Babesia ovis upon microscopic studies, whereas 11 were positive for the presence of Babesia spp. by PCR assay. For the recognition of parasitic DNA, a set of 500bp oligonucleotide was designed by PCR amplification with sequence 18S rRNA gene for B. ovis. The prevalence of babesiosis in small ruminant’s sheep and goat detected by PCR was significantly higher in female animals (28%) than male herds (08%). PCR analysis of the reference samples showed that the detection limit of the PCR assay was 0.01%. Taken together, all data indicated that this PCR assay was a simple, fast, specific detection method for Babesia ovis species in small ruminants compared to other available methods.

Keywords: Babesia ovis, PCR amplification, 18S rRNA, sheep and goat

Procedia PDF Downloads 127

Authors: M. Saidi, I. Djefour, F. Ait Medjber, A. Melouane, A. Gacem

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The objective of this research is to study the physical and mechanical properties and durability of self-compacting mortars prepared by substituting a part of cement up to a percentage of 30% pozzolan according to different Blaine specific surface area (SSB1=7000 cm²/g and SSB=9000 cm²/g)). Order to evaluate durability, mortars were subjected to chemical attacks in various aggressive environments, a solution of a mixture of nitric acid and ammonium nitrate (HNO₃ + NH₄NO₃) and a magnesium sulfate salt solution (MgSO₄)) with a concentration of 10%, for a period of one month. This study is complemented by a comparative study of the durability of mortars elaborated with sulphate resistant cement (SRC). The results show that these mortars develop long-term, mechanical and chemical resistance better than mortars based Portland cement with 5% gypsum (CEM 1) and SRC. We found that the mass losses are lowest in mortars elaborated with pozzolanic cement (30% substitution with SSB₂) in both of chemical attack solutions (3.28% in the solution acid and 1.16% in the salt solution) and the compressive strength gains of 14.68% and 8.5% respectively in the two media. This is due to the action of pozzolan which fixes portlandite to form hydrated calcium silicate (CSH) from the hydration of tricalcic silicate (C₃S).

Keywords: aggressive environments, durability, mechanical strengths, pozzolanic cement, self-compacting mortar

Procedia PDF Downloads 235

Authors: I-Yun Cheng, Min-Yu Chiang, Shwu-Jen Chang, San-Yuan Chen

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Osteoarthritis (OA) is among the most challenging joint diseases, and as far as we know, there is currently no exact and effective cure for it because it has low self-repair ability due to lack of blood vessels and low cell density in articular cartilage. So far, there have been several methods developed to treat cartilage disorder. The most common method is to treat the high molecular weight of hyaluronic acid (HA) injection, but it will degrade after a period of time, so the patients need to inject HA repeatedly. In recent years, self-healing hydrogel has drawn considerable attention because it can recover its initial mechanical properties after damaged and further increase the lifetime of the hydrogel. Here, we aim to develop a self-healable composite hydrogel combined with magnetic microspheres to trigger glutathione(GSH) release for promoting cartilage repair. We use HA-cyclodextrin (CD) as host polymer and poly(acrylic acid)-ferrocene (pAA-Fc) as guest polymer to form the self-healable HA-pAA hydrogel by host and guest interaction where various graft amount of pAA-Fc (pAA:Fc= 1:2, 1:1.5, 1:1, 2:1, 4:1) was conducted to develop different mechanical strength hydrogel. The rheology analysis showed that the 4:1 of pAA-Fc has higher mechanical strength than other formulations. On the other hand, iron oxide nanoparticle, poly(lactic-co-glycolic acid) (PLGA) and polyethyleneimine (PEI) were used to synthesize porous magnetic microspheres via double emulsification water-in-oil-in-water (W/O/W) to increase GSH loading which acted as a reductant to control the hydrogel crosslink density and promote hydrogel self-healing. The results show that the porous magnetic microspheres can be loaded with 70% of GSH and sustained release about 50% of GSH after 24 hours. More importantly, the HA-pAA composite hydrogel can self-heal rapidly within 24 hours when suffering external force destruction by releasing GSH from the magnetic microspheres. Therefore, the developed the HA-pAA composite hydrogel combined with GSH-loaded magnetic microspheres can be in-vivo guided to damaged OA surface for inducing the cartilage repair by controlling the crosslinking of self-healing hydrogel via GSH release.

Keywords: articular cartilage, magnetic microsphere, osteoarthritis, self-healing hydrogel

Procedia PDF Downloads 133

Authors: Abdelkader Basli, Jean-Claude Delaunay, Eric Pedrot, Jean-Michel Mérillon, Jean-Pierre Monti, Khodir Madani, Mohamed Chibane, Tristan Richard

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The antioxidant and antimicrobial activities of Origanum glandulosum collected in Algeria have been studied. Extract was prepared from aerial part of endemic Algerian oregano. The produced extract has been characterized in terms of total phenols (using Folin method), total flavonoid, antioxidant activities (using the DPPH radical scavenging method and ORAC assay) and microbial activity against four bacteria: Streptococcus aureus, Bacillus subtilis, Escherichia coli, Klebsiella pneumoniae one yeast: Candida albicans and one fungi: Aspergillus niger. The results pointed the antioxidant activities of the extract of O. glandulosum and antimicrobial activities against all bacteria and C. Candida, but no effect on A. niger. High performance liquid chromatography combined with mass spectrometry (LC-MS) and nuclear magnetic resonance (LC-NMR) were used to separate and identify the major compounds present in the oregano extract. Rosmarinic acid, globoidnan A and B, lithospermic acid B and three flavonoids were identified.

Keywords: origanum glandulosum, antioxidant, microbial activity, polyphenol, LC-MS, LC-NMR

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Authors: Bo Keun Lee, Doo Yeon Kwon, Ji Hoon Park, Gun Hee Lee, Ji Hye Baek, Heung Jae Chun, Young Joo Koh, Moon Suk Kim

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Thermo-responsive materials are viscoelastic materials that undergo a sol-to-gel phase transition at a specific temperature and many materials have been developed. MPEG-b-PCL (MPC) as a thermo-responsive material contained hydrophilic and hydrophobic segments and it formed an ordered crystalline structure of hydrophobic PCL segments in aqueous solutions. The ordered crystalline structure packed tightly or aggregated and finally induced an aggregated gel through intra- and inter-molecular interactions as a function of temperature. Thus, we introduced anionic and cationic groups into the end positions of the PCL chain to alter the hydrophobicity of the PCL segment. Introducing anionic and cationic groups into the PCL end position altered their solubility by changing the crystallinity and hydrophobicity of the PCL block domains. These results indicated that the properties of the end group in the hydrophobic PCL blockand the balance between hydrophobicity and hydrophilicity affect thermo-responsivebehavior of the copolymers in aqueous solutions. Thus, we concluded that determinant of the temperature-dependent thermo-responsive behavior of MPC depend on the ionic end group in the PCL block. So, we introduced zwitterionic end groups to investigate the thermo-responsive behavior of MPC. Methoxypoly(ethylene oxide) and ε-caprolactone (CL) were randomly copolymerized that introduced varying hydrophobic PCL lengths and an MPC featuring a zwitterionic sulfobetaine (MPC-ZW) at the chain end of the PCL segment. The MPC and MPC-ZW copolymers were obtained formed sol-state at room temperature when prepared as 20-wt% aqueous solutions. The solubility of MPC decreased when the PCL block was increased from molecular weight. The solubilization time of MPC-2.4k was around 20 min and MPC-2.8k, MPC-3.0k increased to 30 min and 1 h, respectively. MPC-3.6k was not solubilized. In case of MPC-ZW 3.6k, However, the zwitterion-modified MPC copolymers were solubilized in 3–5 min. This result indicates that the zwitterionic end group of the MPC-ZW diblock copolymer increased the aqueous solubility of the diblock copolymer even when the length of the hydrophobic PCL segment was increased. MPC and MPC-ZW diblock copolymers that featuring zwitterionic end groups were synthesized successfully. The sol-to-gel phase-transition was formed that specific temperature depend on the length of the PCL hydrophobic segments introduced and on the zwitterion groups attached to the MPC chain end. This result indicated that the zwitterionic end groups reduced the hydrophobicity in the PCL block and changed the solubilization. The MPC-ZW diblock copolymer can be utilized as a potential injectable drug and cell carrier.

Keywords: thermo-responsive material, zwitterionic, hydrophobic, crystallization, phase transition

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Authors: M. Ghasemi, S.Fazelifar

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Abstract Aim: Over-weight is not desirable and has implications for health and in the case of athletes affects performance. Exercise is a strategy used to counteract overweight owing to create a negative energy balance by increasing energy expenditure and influencing appetite regulating hormones. Interestingly, recent studies have revealed inhibitory effects of exercise on the hunger associated with these hormones in healthy subjects Neuropeptide Y(NPY) is a 36 amino acid protein that is a powerful stimulant appetite. NPY is an important central orexigenic hormone predominantly produced by the hypothalamus, and recently found to be secreted in adipose tissue. This neurotransmitter is secreted in the brain and autonomic nervous system. On the other hand, research has shown that thyme in addition to various properties, also affects the appetite. The purpose of this study was to determine Effects of eight weeks endurance training and thyme consumption on neuropeptide Y in untrained men. Methodology: 36 Healthy untrained men (mean body weight 78.25±3.2 kg, height 176±6.8 cm, age 34.32±4.54 years and BMI 29.1±4.3 kg/m2) voluntarily participated in this study . Subjects were randomly divided into four groups: 1. control, 2. Endurance training, 3. Thyme 4. Endurance training + Thyme. Amount of 10cc Blood sampling were obtained pre-test and post-test (after 8 weeks). The taken blood samples were centrifuged at 1500 × g for 15 min then plasma was stored at -20 °C until analysis. Endurance training consisted three session per week with 60% -75% of reserve heart rate for eight weeks. Exclusion criteria were history of gastrointestinal, endocrine, cardiovascular or psychological disease, and consuming any supplementation, alcohol and tobacco products. Descriptive statistics including means, standard deviations, and ranges were calculated for all measures. K-S test to determine the normality of the data and analysis of variance for repeated measures was used to analyze the data. A significant difference in the p<0/05 accepted. Results: Results showed that aerobic training significantly reduced body weight, body mass index, percent body fat, but significant increase observed in maximal oxygen consumption level (p ≤ 0/05). The neuropeptide Y levels were significantly increased after exercise. Analysis of data determined that there was no significant difference between the four groups. Conclusion: Appetite control plays a critical role in the competition between energy consumption and energy expenditure. The results of this study showed that endurance training and thyme consumption can be cause improvement in physiological parameters such as increasing aerobic capacity, reduction of fat mass and improve body composition in untrained men.

Keywords: Endurance training, neuropeptide Y, thyme, untrained men

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Authors: Kuheli Chakrabarty, Animesh Ghosh, Atanu Roy, Gourab Kanti Das

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Aldol reaction is an important class of carbon-carbon bond forming reactions. One of the popular ways to impose asymmetry in aldol reaction is the introduction of chiral auxiliary that binds the approaching reactants and create dissymmetry in the reaction environment, which finally evolves to enantiomeric excess in the aldol products. The last decade witnesses the usage of natural amino acids as chiral auxiliary to control the stereoselectivity in various carbon-carbon bond forming processes. In this context, L-proline was found to be an effective organocatalyst in asymmetric aldol additions. In last few decades the use of water as solvent or co-solvent in asymmetric organocatalytic reaction is increased sharply. Simple amino acids like L-proline does not catalyze asymmetric aldol reaction in aqueous medium not only that, In organic solvent medium high catalytic loading (~30 mol%) is required to achieve moderate to high asymmetric induction. In this context, huge efforts have been made to modify L-proline and 4-hydroxy-L-proline to prepare organocatalyst for aqueous medium asymmetric aldol reaction. Here, we report the result of our DFT calculations on asymmetric aldol reaction of benzaldehyde, p-NO2 benzaldehyde and t-butyraldehyde with a number of ketones using L-proline hydrazide as organocatalyst in wet solvent free condition. Gaussian 09 program package and Gauss View program were used for the present work. Geometry optimizations were performed using B3LYP hybrid functional and 6-31G(d,p) basis set. Transition structures were confirmed by hessian calculation and IRC calculation. As the reactions were carried out in solvent free condition, No solvent effect were studied theoretically. Present study has revealed for the first time, the direct involvement of two water molecules in the aldol transition structures. In the TS, the enamine and the aldehyde is connected through hydrogen bonding by the assistance of two intervening water molecules forming a supramolecular network. Formation of this type of supramolecular assembly is possible due to the presence of protonated -NH2 group in the L-proline hydrazide moiety, which is responsible for the favorable entropy contribution to the aldol reaction. It is also revealed from the present study that, water assisted TS is energetically more favorable than the TS without involving any water molecule. It can be concluded from this study that, insertion of polar group capable of hydrogen bond formation in the L-proline skeleton can lead to a favorable aldol reaction with significantly high enantiomeric excess in wet solvent free condition by reducing the activation barrier of this reaction.

Keywords: aldol reaction, DFT, organocatalysis, transition structure

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