Search results for: RNA binding
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 842

Search results for: RNA binding

512 In Silico Study of the Biological and Pharmacological Activity of Nigella sativa

Authors: Ammar Ouahab, Meriem Houichi , Sanna Mihoubi

Abstract:

Background: Nigella sativa is an annual flowering plant, belongs to the Ranunculaceae family. It has many pharmacological activities such as anti-inflammatory; anti-bacterial; anti-hepatotoxic activities etc. Materials: In order to predict the pharmacological activity of Nigella Sativa’s compounds, some web based servers were used, namely, PubChem, Molinspiration, ADMET-SAR, PASS online and PharMapper. In addition to that, AutoDOCK was used to investigate the different molecular interactions between the selected compounds and their target proteins. Results: All compounds displayed a stable interaction with their targets and satisfactory binding energies, which means that they are active on their targets. Conclusion: Nigella sativa is an effective medicinal plant that has several ethno-medical uses; the latter uses are proven herein via an in-silico study of their pharmacological activities.

Keywords: Nigella sativa, AutoDOCK, PubChem, Molinspiration, ADMET-SAR, PharMapper, PASS online server, docking

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511 The Structural Alteration of DNA Native Structure of Staphylococcus aureus Bacteria by Designed Quinoxaline Small Molecules Result in Their Antibacterial Properties

Authors: Jeet Chakraborty, Sanjay Dutta

Abstract:

Antibiotic resistance by bacteria has proved to be a severe threat to mankind in recent times, and this fortifies an urgency to design and develop potent antibacterial small molecules/compounds with nonconventional mechanisms than the conventional ones. DNA carries the genetic signature of any organism, and bacteria maintain their genomic DNA inside the cell in a well-regulated compact form with the help of various nucleoid associated proteins like HU, HNS, etc. These proteins control various fundamental processes like gene expression, replication, etc., inside the cell. Alteration of the native DNA structure of bacteria can lead to severe consequences in cellular processes inside the bacterial cell that ultimately result in the death of the organism. The change in the global DNA structure by small molecules initiates a plethora of cellular responses that have not been very well investigated. Echinomycin and Triostin-A are biologically active Quinoxaline small molecules that typically consist of a quinoxaline chromophore attached with an octadepsipeptide ring. They bind to double-stranded DNA in a sequence-specific way and have high activity against a wide variety of bacteria, mainly against Gram-positive ones. To date, few synthetic quinoxaline scaffolds were synthesized, displaying antibacterial potential against a broad scale of pathogenic bacteria. QNOs (Quinoxaline N-oxides) are known to target DNA and instigate reactive oxygen species (ROS) production in bacteria, thereby exhibiting antibacterial properties. The divergent role of Quinoxaline small molecules in medicinal research qualifies them for the evaluation of their antimicrobial properties as a potential candidate. The previous study from our lab has given new insights on a 6-nitroquinoxaline derivative 1d as an intercalator of DNA, which induces conformational changes in DNA upon binding.7 The binding event observed was dependent on the presence of a crucial benzyl substituent on the quinoxaline moiety. This was associated with a large induced CD (ICD) appearing in a sigmoidal pattern upon the interaction of 1d with dsDNA. The induction of DNA superstructures by 1d at high Drug:DNA ratios was observed that ultimately led to DNA condensation. Eviction of invitro-assembled nucleosome upon treatment with a high dose of 1d was also observed. In this work, monoquinoxaline derivatives of 1d were synthesized by various modifications of the 1d scaffold. The set of synthesized 6-nitroquinoxaline derivatives along with 1d were all subjected to antibacterial evaluation across five different bacteria species. Among the compound set, 3a displayed potent antibacterial activity against Staphylococcus aureus bacteria. 3a was further subjected to various biophysical studies to check whether the DNA structural alteration potential was still intact. The biological response of S. aureus cells upon treatment with 3a was studied using various cell biology processes, which led to the conclusion that 3d can initiate DNA damage in the S. aureus cells. Finally, the potential of 3a in disrupting preformed S.aureus and S.epidermidis biofilms was also studied.

Keywords: DNA structural change, antibacterial, intercalator, DNA superstructures, biofilms

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510 Investigation of Chlorophylls a and b Interaction with Inner and Outer Surfaces of Single-Walled Carbon Nanotube Using Molecular Dynamics Simulation

Authors: M. Dehestani, M. Ghasemi-Kooch

Abstract:

In this work, adsorption of chlorophylls a and b pigments in aqueous solution on the inner and outer surfaces of single-walled carbon nanotube (SWCNT) has been studied using molecular dynamics simulation. The linear interaction energy algorithm has been used to calculate the binding free energy. The results show that the adsorption of two pigments is fine on the both positions. Although there is the close similarity between these two pigments, their interaction with the nanotube is different. This result is useful to separate these pigments from one another. According to interaction energy between the pigments and carbon nanotube, interaction between these pigments-SWCNT on the inner surface is stronger than the outer surface. The interaction of SWCNT with chlorophylls phytol tail is stronger than the interaction of SWCNT with porphyrin ring of chlorophylls.

Keywords: adsorption, chlorophyll, interaction, molecular dynamics simulation, nanotube

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509 An Investigation of Tetraspanin Proteins’ Role in UPEC Infection

Authors: Fawzyah Albaldi

Abstract:

Urinary tract infections (UTIs) are the most prevalent of infectious diseases and > 80% are caused by uropathogenic E. coli (UPEC). Infection occurs following adhesion to urothelial plaques on bladder epithelial cells, whose major protein constituent are the uroplakins (UPs). Two of the four uroplakins (UPIa and UPIb) are members of the tetraspanin superfamily. The UPEC adhesin FimH is known to interact directly with UPIa. Tetraspanins are a diverse family of transmembrane proteins that generally act as “molecular organizers” by binding different proteins and lipids to form tetraspanin enriched microdomains (TEMs). Previous work by our group has shown that TEMs are involved in the adhesion of many pathogenic bacteria to human cells. Adhesion can be blocked by tetraspanin-derived synthetic peptides, suggesting that tetraspanins may be valuable drug targets. In this study, we investigate the role of tetraspanins in UPEC adherence to bladder epithelial cells. Human bladder cancer cell lines (T24, 5637, RT4), commonly used as in-vitro models to investigate UPEC infection, along with primary human bladder cells, were used in this project. The aim was to establish a model for UPEC adhesion/infection with the objective of evaluating the impact of tetraspanin-derived reagents on this process. Such reagents could reduce the progression of UTI, particularly in patients with indwelling catheters. Tetraspanin expression on the bladder cells was investigated by q-PCR and flow cytometry, with CD9 and CD81 generally highly expressed. Interestingly, despite these cell lines being used by other groups to investigate FimH antagonists, uroplakin proteins (UPIa, UPIb and UPIII) were poorly expressed at the cell surface, although some were present intracellularly. Attempts were made to differentiate the cell lines, to induce cell surface expression of these UPs, but these were largely unsuccessful. Pre-treatment of bladder epithelial cells with anti-CD9 monoclonal antibody significantly decreased UPEC infection, whilst anti-CD81 had no effects. A short (15aa) synthetic peptide corresponding to the large extracellular region (EC2) of CD9 also significantly reduced UPEC adherence. Furthermore, we demonstrated specific binding of that fluorescently tagged peptide to the cells. CD9 is known to associate with a number of heparan sulphate proteoglycans (HSPGs) that have also been implicated in bacterial adhesion. Here, we demonstrated that unfractionated heparin (UFH)and heparin analogs significantly inhibited UPEC adhesion to RT4 cells, as did pre-treatment of the cells with heparinases. Pre-treatment with chondroitin sulphate (CS) and chondroitinase also significantly decreased UPEC adherence to RT4 cells. This study may shed light on a common pathogenicity mechanism involving the organisation of HSPGs by tetraspanins. In summary, although we determined that the bladder cell lines were not suitable to investigate the role of uroplakins in UPEC adhesion, we demonstrated roles for CD9 and cell surface proteoglycans in this interaction. Agents that target these may be useful in treating/preventing UTIs.

Keywords: UTIs, tspan, uroplakins, CD9

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508 An Electrochemical DNA Biosensor Based on Oracet Blue as a Label for Detection of Helicobacter pylori

Authors: Saeedeh Hajihosseini, Zahra Aghili, Navid Nasirizadeh

Abstract:

An innovative method of a DNA electrochemical biosensor based on Oracet Blue (OB) as an electroactive label and gold electrode (AuE) for detection of Helicobacter pylori, was offered. A single–stranded DNA probe with a thiol modification was covalently immobilized on the surface of the AuE by forming an Au–S bond. Differential pulse voltammetry (DPV) was used to monitor DNA hybridization by measuring the electrochemical signals of reduction of the OB binding to double– stranded DNA (ds–DNA). Our results showed that OB–based DNA biosensor has a decent potential for detection of single–base mismatch in target DNA. Selectivity of the proposed DNA biosensor was further confirmed in the presence of non–complementary and complementary DNA strands. Under optimum conditions, the electrochemical signal had a linear relationship with the concentration of the target DNA ranging from 0.3 nmol L-1 to 240.0 nmol L-1, and the detection limit was 0.17 nmol L-1, whit a promising reproducibility and repeatability.

Keywords: DNA biosensor, oracet blue, Helicobacter pylori, electrode (AuE)

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507 Effect of Resveratrol and Ascorbic Acid on the Stability of Alfa-Tocopherol in Whey Protein Isolate Stabilized O/W Emulsions

Authors: Lei Wang, Yingzhou Ni, Amr M. Bakry, Hao Cheng, Li Liang

Abstract:

Food proteins have been widely used as carrier materials because of their multiple functional properties. In this study, alfa-tocopherol was encapsulated in the oil phase of an oil-in-water emulsion stabilized with whey protein isolate (WPI). The influence of WPI concentration and resveratrol or ascorbic acid on the decomposition of alfa-tocopherol in the emulsion during storage is discussed. Decomposition decreased as WPI concentrations increased. Decomposition was delayed at ascorbic acid/WPI molar ratios lower than 5 but was promoted at higher ratios. Resveratrol partitioned into the oil-water interface by binding to WPI and its cis-isomer is believed to have contributed most of the protective effect of this polyphenol. These results suggest the possibility of using the emulsifying and ligand-binging properties of WPI to produce carriers for simultaneous encapsulation of alfa-tocopherol and resveratrol in a single emulsion system.

Keywords: stability, alfa-tocopherol, resveratrol, whey protein isolate

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506 Foslip Loaded and CEA-Affimer Functionalised Silica Nanoparticles for Fluorescent Imaging of Colorectal Cancer Cells

Authors: Yazan S. Khaled, Shazana Shamsuddin, Jim Tiernan, Mike McPherson, Thomas Hughes, Paul Millner, David G. Jayne

Abstract:

Introduction: There is a need for real-time imaging of colorectal cancer (CRC) to allow tailored surgery to the disease stage. Fluorescence guided laparoscopic imaging of primary colorectal cancer and the draining lymphatics would potentially bring stratified surgery into clinical practice and realign future CRC management to the needs of patients. Fluorescent nanoparticles can offer many advantages in terms of intra-operative imaging and therapy (theranostic) in comparison with traditional soluble reagents. Nanoparticles can be functionalised with diverse reagents and then targeted to the correct tissue using an antibody or Affimer (artificial binding protein). We aimed to develop and test fluorescent silica nanoparticles and targeted against CRC using an anti-carcinoembryonic antigen (CEA) Affimer (Aff). Methods: Anti-CEA and control Myoglobin Affimer binders were subcloned into the expressing vector pET11 followed by transformation into BL21 Star™ (DE3) E.coli. The expression of Affimer binders was induced using 0.1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG). Cells were harvested, lysed and purified using nickle chelating affinity chromatography. The photosensitiser Foslip (soluble analogue of 5,10,15,20-Tetra(m-hydroxyphenyl) chlorin) was incorporated into the core of silica nanoparticles using water-in-oil microemulsion technique. Anti-CEA or control Affs were conjugated to silica nanoparticles surface using sulfosuccinimidyl-4-(N-maleimidomethyl) cyclohexane-1-carboxylate (sulfo SMCC) chemical linker. Binding of CEA-Aff or control nanoparticles to colorectal cancer cells (LoVo, LS174T and HC116) was quantified in vitro using confocal microscopy. Results: The molecular weights of the obtained band of Affimers were ~12.5KDa while the diameter of functionalised silica nanoparticles was ~80nm. CEA-Affimer targeted nanoparticles demonstrated 9.4, 5.8 and 2.5 fold greater fluorescence than control in, LoVo, LS174T and HCT116 cells respectively (p < 0.002) for the single slice analysis. A similar pattern of successful CEA-targeted fluorescence was observed in the maximum image projection analysis, with CEA-targeted nanoparticles demonstrating 4.1, 2.9 and 2.4 fold greater fluorescence than control particles in LoVo, LS174T, and HCT116 cells respectively (p < 0.0002). There was no significant difference in fluorescence for CEA-Affimer vs. CEA-Antibody targeted nanoparticles. Conclusion: We are the first to demonstrate that Foslip-doped silica nanoparticles conjugated to anti-CEA Affimers via SMCC allowed tumour cell-specific fluorescent targeting in vitro, and had shown sufficient promise to justify testing in an animal model of colorectal cancer. CEA-Affimer appears to be a suitable targeting molecule to replace CEA-Antibody. Targeted silica nanoparticles loaded with Foslip photosensitiser is now being optimised to drive photodynamic killing, via reactive oxygen generation.

Keywords: colorectal cancer, silica nanoparticles, Affimers, antibodies, imaging

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505 Preparation of Geopolymer Cements from Tunisian Illito-Kaolinitic Clay Mineral

Authors: N. Hamdi, E. Srasra

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In this work geopolymer cement are synthesized from Tunisian (illito-kaolinitic) clay. This product can be used as binding material in place of cement Portland. The clay fractions used were characterized with physico-chemical and thermal analyses. The clays materials react with alkaline solution (10, 14 and 18 mol(NaOH)/L) in order to produce geopolymer cements whose pastes were characterized by determining their water adsorption and compressive strength. The compressive strength of the hardened geopolymer cement paste samples aged 28 days attained its highest value (32.3MPa) around 950°C for NaOH concentration of 14M. The water adsorption value of the prepared samples decreased with increasing the calcination temperature of clay fractions. It can be concluded that the most suitable temperature for the calcination of illitio-kaolinitic clays in view of producing geopolymer cements is around 950°C.

Keywords: compressive strength, geopolymer cement, illitio-kaolinitic clay, mineral

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504 Recovery of Post-Consumer PET Bottles in a Composite Material Preparation

Authors: Rafenomananjara Tsinjo Nirina, Tomoo Sekito, Andrianaivoravelona Jaconnet Oliva

Abstract:

Manufacturing a composite material from post-consumer bottles is an interesting outlet since Madagascar is still facing the challenges of managing plastic waste on the one hand and appropriate waste treatment facilities are not yet developed on the other hand. New waste management options are needed to divert End-Of-Life (EOL) soft plastic wastes from landfills and incineration. Waste polyethylene terephthalate (PET) bottles might be considered as a valuable resource and recovered into polymer concrete. The methodology is easy to implement and appropriate to the local context in Madagascar. This approach will contribute to the production of ecological building materials that might be profitable for the environment and the construction sector. This work aims to study the feasibility of using the post-consumer PET bottles as an alternative binding agent instead of the conventional Portland cement and water. Then, the mechanical and physical properties of the materials were evaluated.

Keywords: PET recycling, polymer concrete, ecological building materials, pollution mitigation

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503 Flexible Polyaniline-Based Composite Films for High-Performance Super Capacitors

Authors: A. Khosrozadeh, M. A. Darabi, M. Xing, Q. Wang

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Fabrication of a high-performance supercapacitor (SC) using a flexible cellulose-based composite film of polyaniline (PANI), reduced graphene oxide (RGO), and silver nanowires (AgNWs) is reported. The flexibility, high capacitive behaviour, and cyclic stability of the entire device make it a good candidate for wearable SCs. The results show that a capacitance as high as 73.4 F/g (1.6 F/cm2) at a discharge rate of 1.1 A/g is achieved by the device. In addition, the SC demonstrates a power density up to 468.8 W/kg and an energy density up to 5.1 wh/kg. The flexibility of the composite film is attributed to the binding effect of cellulose fibers as well as reinforcing effect of AgNWs. The excellent electrochemical performance of the device is found to be owing to the synergistic effect between PANI/RGO/AgNWs ternary in a cushiony cellulose matrix and porous structure of the composite.

Keywords: cellulose, polyaniline, reduced graphene oxide, silver, super capacitor

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502 Exploring Structure of Human Chromosomes Using Fluorescence Lifetime Imaging

Authors: A. Bhartiya, S. Botchway, M. Yusuf, I. Robinson

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Chromatin condensation is maintained by DNA-based proteins and some divalent cations (Mg²⁺, Ca²⁺, etc.). Condensation process during cell division maintains structural and functional organizations of chromosomes by transferring genetic information correctly to daughter cells. Fluorescence Lifetime Imaging (FLIM) technique measures the fluorescence decay of fixed human chromosomes by calculating the lifetime of fluorophores at a pixel x of the arrival of each photon as a function of time delay t, following excitation with a laser pulse. Fixed metaphase human chromosomes were labelled with DNA-binding dye, DAPI and later DAPI fluorescence lifetime measured using multiphoton microscopy. 5 out of 23 pairs of human chromosomes shown shorter lifetime at the centromere region, differentiating proportion of compaction along the length of chromosomes. Different lifetime was observed in a condensed and de-condensed chromosome. It clearly indicates the involvement of divalent cations in the process of condensation.

Keywords: divalent cations, FLIM (Fluorescence Lifetime Imaging), human chromosomes, multiphoton microscopy

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501 Inhibition of α-Glucosidase and Xanthine Oxidase by Curcumin and Its Analogs

Authors: Jung-Feng Hsieh, Chu Ze Chen

Abstract:

Curcumin is the main active compound of turmeric that can inhibit the activities of α-glucosidase and xanthine oxidase (XO). α-Glucosidase and XO inhibitors are widely used to treat patients with diabetes mellitus and gout, respectively; therefore, the objective of this research was to evaluate the inhibitory activities of curcumin and its analogs against α-glucosidase and XO. Our results demonstrated that CM-F had the strongest antioxidant activity with a half-maximal effective concentration (EC50) of 9.39 ± 0.16 μM, which was superior to vitamin E (EC50=17.03 ± 0.09 μM). CM-F also exhibited potent inhibitory activity against XO with an IC50 value of 6.14 ± 0.38 μM and enzyme kinetic results revealed competitive inhibition of XO. We also found that CM-1 and CM-2 inhibited α-glucosidase with IC50 values of 21.06 ± 0.92 μM and 5.95 ± 0.09 μM, respectively, and kinetic studies indicated that both CM-1 and CM-2 are mixed competitive inhibitors of α-glucosidase. Furthermore, docking simulation identified five hydrogen bonds between XO and CM-F; however, only one and two hydrogen bonds are involved in CM-1 and CM-2 binding to α-glucosidase, respectively. Accordingly, curcumin and its analogs have the potential to be used in the treatment of patients with diabetes mellitus and gout.

Keywords: curcumin, α-glucosidase, inhibitor, xanthine oxidase

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500 Performance of Armchair Graphene Nanoribbon Resonant Tunneling Diode under Uniaxial Strain

Authors: Milad Zoghi, M. Zahangir Kabir

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Performance of armchair graphene nanoribbon (AGNR) resonant tunneling diodes (RTD) alter if they go under strain. This may happen due to either using stretchable substrates or real working conditions such as heat generation. Therefore, it is informative to understand how mechanical deformations such as uniaxial strain can impact the performance of AGNR RTDs. In this paper, two platforms of AGNR RTD consist of width-modified AGNR RTD and electric-field modified AGNR RTD are subjected to both compressive and tensile uniaxial strain ranging from -2% to +2%. It is found that characteristics of AGNR RTD markedly change under both compressive and tensile strain. In particular, peak to valley ratio (PVR) can be totally disappeared upon strong enough strain deformation. Numerical tight binding (TB) coupled with Non-Equilibrium Green's Function (NEGF) is derived for this study to calculate corresponding Hamiltonian matrices and transport properties.

Keywords: armchair graphene nanoribbon, resonant tunneling diode, uniaxial strain, peak to valley ratio

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499 Previously Undescribed Cardiac Abnormalities in Two Unrelated Autistic Males with Causative Variants in CHD8

Authors: Mariia A. Parfenenko, Ilya S. Dantsev, Sergei V. Bochenkov, Natalia V. Vinogradova, Olga S. Groznova, Victoria Yu. Voinova

Abstract:

Introduction: Autism is the most common neurodevelopmental disorder. Autism is characterized by difficulties in social interaction and adherence to stereotypic behavioral patterns and frequently co-occurs with epilepsy, intellectual disabilities, connective tissue disorders, and other conditions. CHD8 codes for chromodomain-helicase-DNA-binding protein 8 - a chromatin remodeler that regulates cellular proliferation and neurodevelopment in embryogenesis. CHD8 is one of the genes most frequently involved in autism. Patients and methods: 2 unrelated male patients, P3 and P12, aged 3 and 12 years old, underwent whole genome sequencing, which determined that they both had different likely pathogenic variants, both previously undescribed in literature. Sanger sequencing later determined that P12 inherited the variant from his affected mother. Results: P3 and P12 presented with autism, a developmental delay, ataxia, sleep disorders, overgrowth, and macrocephaly, as well as other clinical features typically present in patients with causative variants in CHD8. The mother of P12 also has autistic traits, as well as ataxia, hypotonia, sleep disorders, and other symptoms. However, P3 and P12 also have different cardiac abnormalities. P3 had signs of a repolarization disorder: a flattened T wave in the III and aVF derivations and a negative T wave in the V1-V2 derivations. He also had structural valve anomalies with associated regurgitation, local contractility impairment of the left ventricular, and diastolic dysfunction of the right ventricle. Meanwhile, P12 had Wolff-Parkinson-White syndrome and underwent radiofrequency ablation at the age of 2 years. At the time of observation, P12 had mild sinus arrhythmia and an incomplete right bundle branch block, as well as arterial hypertension. Discussion: Cardiac abnormalities were not previously reported in patients with causative variants in CHD8. The underlying mechanism for the formation of those abnormalities is currently unknown. However, the two hypotheses are either a disordered interaction with CHD7 – another chromodomain remodeler known to be directly involved in the cardiophenotype of CHARGE syndrome – a rare condition characterized by coloboma, heart defects and growth abnormalities, or the disrupted functioning of CHD8 as an A-Kinase Anchoring Protein, which are known to modulate cardiac function. Conclusion: We observed 2 unrelated autistic males with likely pathogenic variants in CHD8 that presented with typical symptoms of CHD8-related neurodevelopmental disorder, as well as cardiac abnormalities. Cardiac abnormalities have, until now, been considered uncharacteristic for patients with causative variants in CHD8. Further accumulation of data, including experimental evidence of the involvement of CHD8 in heart formation, will elucidate the mechanism underlying the cardiophenotype of those patients. Acknowledgements: Molecular genetic testing of the patients was made possible by the Charity Fund for medical and social genetic aid projects «Life Genome.»

Keywords: autism spectrum disorders, chromodomain-helicase-DNA-binding protein 8, neurodevelopmental disorder, cardio phenotype

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498 Loss in Efficacy of Viscoelastic Ionic Liquid Surfactants under High Salinity during Surfactant Flooding

Authors: Shilpa K. Nandwani, Mousumi Chakraborty, Smita Gupta

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When selecting surfactants for surfactant flooding during enhanced oil recovery, the most important criteria is that the surfactant system should reduce the interfacial tension between water and oil to ultralow values. In the present study, a mixture of ionic liquid surfactant and commercially available binding agent sodium tosylate has been used as a surfactant mixture. Presence of wormlike micelles indicates the possibility of achieving ultralow interfacial tension. Surface tension measurements of the mixed surfactant system have been studied. The emulsion size distribution of the mixed surfactant system at varying salinities has been studied. It has been found that at high salinities the viscoelastic surfactant system loses their efficacy and degenerate. Hence the given system may find application in low salinity reservoirs, providing good mobility to the flood during tertiary oil recovery process.

Keywords: ionic liquis, interfacial tension, Na-tosylate, viscoelastic surfactants

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497 Detection of Nanotoxic Material Using DNA Based QCM

Authors: Juneseok You, Chanho Park, Kuehwan Jang, Sungsoo Na

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Sensing of nanotoxic materials is strongly important, as their engineering applications are growing recently and results in that nanotoxic material can harmfully influence human health and environment. In current study we report the quartz crystal microbalance (QCM)-based, in situ and real-time sensing of nanotoxic-material by frequency shift. We propose the in situ detection of nanotoxic material of zinc oxice by using QCM functionalized with a taget-specific DNA. Since the mass of a target material is comparable to that of an atom, the mass change caused by target binding to DNA on the quartz electrode is so small that it is practically difficult to detect the ions at low concentrations. In our study, we have demonstrated the in-situ and fast detection of zinc oxide using the quartz crystal microbalance (QCM). The detection was derived from the DNA hybridization between the DNA on the quartz electrode. The results suggest that QCM-based detection opens a new avenue for the development of a practical water-testing sensor.

Keywords: nanotoxic material, qcm, frequency, in situ sensing

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496 FTIR Characterization of EPS Ligands from Mercury Resistant Bacterial Isolate, Paenibacillus jamilae PKR1

Authors: Debajit Kalita, Macmillan Nongkhlaw, S. R. Joshi

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Mercury (Hg) is a highly toxic heavy metal released both from naturally occurring volcanoes and anthropogenic activities like alkali and mining industries as well as biomedical wastes. Exposure to mercury is known to affect the nervous, gastrointestinal and renal systems. In the present study, a bacterial isolate identified using 16S rRNA marker as Paenibacillus jamilae PKR1 isolated from India’s largest sandstone-type uranium deposits, containing an average of 0.1% U3O8, was found to be resistance to Hg contamination under culture conditions. It showed strong hydrophobicity as revealed by SAT, MATH, PAT, SAA adherence assays. The Fourier Transform Infrared (FTIR) spectra showed the presence of hydroxyl, amino and carboxylic functional groups on the cell surface EPS which are known to contribute in the binding of metals. It is proposed that the characterized isolate tolerating up to 4.0mM of mercury provides scope for its application in bioremediation of mercury from contaminated sites.

Keywords: mercury, Domiasiat, uranium, paenibacillus jamilae, hydrophobicity, FTIR

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495 Impact of Flavor on Food Product Quality, A Case Study of Vanillin Stability during Biscuit Preparation

Authors: N. Yang, R. Linforth, I. Fisk

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The influence of food processing and choice of flavour solvent was investigated using biscuits prepared with vanillin flavour as an example. Powder vanillin either was added directly into the dough or dissolved into flavour solvent then mixed into the dough. The impact of two commonly used flavour solvents on food quality was compared: propylene glycol (PG) or triacetin (TA). The analytical approach for vanillin detection was developed by chromatography (HPLC-PDA), and the standard extraction method for vanillin was also established. The results indicated the impact of solvent choice on vanillin level during biscuit preparation. After baking, TA as a more heat resistant solvent retained more vanillin than PG, so TA is a better solvent for products that undergo a heating process. The results also illustrated the impact of mixing and baking on vanillin stability in the matrices. The average loss of vanillin was 33% during mixing and 13% during baking, which indicated that the binding of vanillin to fat or flour before baking might cause larger loss than evaporation loss during baking.

Keywords: biscuit, flavour stability, food quality, vanillin

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494 Joint Physical Custody: Lessons from the European Union

Authors: Katarzyna Kamińska

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When thinking about custodial arrangements after divorce or separation, there has been a shift from sole custody, particularly maternal preference, to joint physical custody. In many Western countries, an increasing of children with separated parents have joint physical custody, which is believed to be in the best interests of the child, as children can maintain personal relations and direct contact with both parents on a regular basis. The aim of the article is to examine joint physical custody, both from the perspective of the binding legal instruments that are relevant to joint physical custody, the Principles of European Family Law drafted by the CEFL, as well as the international research on this matter. The thesis underlying this paper is that joint physical custody is in itself neither good nor bad, and it depends on how the arrangements are managed by the parents. The paper includes a reflection on joint physical custody in the face of the COVID-19 crisis. The results indicate that in normal circumstances, joint physical custody demands broad communication, and now it times of crisis, we need over-communication about children and plans. Only a very tight and coordinated co-parenting plan make the whole family safer.

Keywords: joint physical custody, co-parenting, child welfare, COVID-19

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493 AI-Based Technologies in International Arbitration: An Exploratory Study on the Practicability of Applying AI Tools in International Arbitration

Authors: Annabelle Onyefulu-Kingston

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One of the major purposes of AI today is to evaluate and analyze millions of micro and macro data in order to determine what is relevant in a particular case and proffer it in an adequate manner. Microdata, as far as it relates to AI in international arbitration, is the millions of key issues specifically mentioned by either one or both parties or by their counsels, arbitrators, or arbitral tribunals in arbitral proceedings. This can be qualifications of expert witness and admissibility of evidence, amongst others. Macro data, on the other hand, refers to data derived from the resolution of the dispute and, consequently, the final and binding award. A notable example of this includes the rationale of the award and specific and general damages awarded, amongst others. This paper aims to critically evaluate and analyze the possibility of technological inclusion in international arbitration. This research will be imploring the qualitative method by evaluating existing literature on the consequence of applying AI to both micro and macro data in international arbitration, and how this can be of assistance to parties, counsels, and arbitrators.

Keywords: AI-based technologies, algorithms, arbitrators, international arbitration

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492 Promissing Antifungal Chitinase from Marine Strain of Bacillus

Authors: Ben Amar Cheba, Taha Ibrahim Zaghloul, Mohamad Hisham El-Massry, Ahmad Rafik El-Mahdy

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Seventy two bacterial strains with ability to degrade chitin were isolated during a screening program. One of the most potent isolates (strain R2) was identified as Bacillus sp. using conventional methods as well as 16S rRNA technique and submitted in the Gen Bank sequence database as Bacillus sp. R2 with a given accession number DQ 923161. This strain was able to produce high levels of extracellular chitinase. The chitinase of Bacillus sp. R2 hydrolyzed several chitinous substrates preferentially and showed a maximum activity toward the β chitin such as Calmar pen and squid bone chitins with the folds 1.47 and 1.23 respectively. The enzyme also exhibited a substrate binding capacity of more than 70% for squid chitin, shrimp shell colloidal chitin, chitosan and prawn shell chitin. The chitinase showed a moderate antifungal activity against many phytopathogenic fungi such as Aspergillus niger, A. flavus, Penicillium degitatum and Fusarium calmorum.This strain could be a suitable candidate for chitinase production on an industrial scale for using as promising antifungal biopestecide.

Keywords: antifungal activity, Bacillus sp. R2, chitinase, substrate specificity

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491 Investigation of Biofilm Formation in Clinical Strains of Klebsiella pneumoniae and Klebsiella rhinoscleromatis

Authors: Gulcan Sahal, Nermin Hande Avcioglu, Isil Seyis Bilkay

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Klebsiella species which are natural colonizers of human upper respiratory and human gastrointestinal tracts are also responsible for every reoccurring nosocomial infections by means of having ability to form slimy layers known as biofilm on many surfaces. Therefore, in this study, investigation of biofilm formation in K. pneumoniae and K. rhinoscleromatis and examination of each Klebsiella strains’ clinical information in the light of their biofilm formation results were aimed. In this respect, biofilm formation of Klebsiella strains was analyzed via crystal violet binding assay. According to our results, biofilm formation levels of K. pneumoniae and K. rhinoscleromatis strains were different from each other. Additionally, in comparison to K. rhinoscleromatis strains, K. pneumoniae was observed to include higher amounts of strong biofilm forming strains. Besides, it was also seen that clinical information of patients from which strong biofilm forming Klebsiella strains were isolated were similar to each other. Our results indicate that there should be more precautions against K. pneumoniae which includes higher amount of strong biofilm forming strains.

Keywords: biofilm formation, Klebsiella pneumoniae, Klebsiella rhinoscleromatis, biosystems engineering

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490 Biophysical and Structural Characterization of Transcription Factor Rv0047c of Mycobacterium Tuberculosis H37Rv

Authors: Md. Samsuddin Ansari, Ashish Arora

Abstract:

Every year 10 million people fall ill with one of the oldest diseases known as tuberculosis, caused by Mycobacterium tuberculosis. The success of M. tuberculosis as a pathogen is because of its ability to persist in host tissues. Multidrug resistance (MDR) mycobacteria cases increase every day, which is associated with efflux pumps controlled at the level of transcription. The transcription regulators of MDR transporters in bacteria belong to one of the following four regulatory protein families: AraC, MarR, MerR, and TetR. Phenolic acid decarboxylase repressor (PadR), like a family of transcription regulators, is closely related to the MarR family. Phenolic acid decarboxylase repressor (PadR) was first identified as a transcription factor involved in the regulation of phenolic acid stress response in various microorganisms (including Mycobacterium tuberculosis H37Rv). Recently research has shown that the PadR family transcription factors are global, multifunction transcription regulators. Rv0047c is a PadR subfamily-1 protein. We are exploring the biophysical and structural characterization of Rv0047c. The Rv0047 gene was amplified by PCR using the primers containing EcoRI and HindIII restriction enzyme sites cloned in pET-NH6 vector and overexpressed in DH5α and BL21 (λDE3) cells of E. coli following purification with Ni2+-NTA column and size exclusion chromatography. We did DSC to know the thermal stability; the Tm (transition temperature) of protein is 55.29ºC, and ΔH (enthalpy change) of 6.92 kcal/mol. Circular dichroism to know the secondary structure and conformation and fluorescence spectroscopy for tertiary structure study of protein. To understand the effect of pH on the structure, function, and stability of Rv0047c we employed spectroscopy techniques such as circular dichroism, fluorescence, and absorbance measurements in a wide range of pH (from pH-2.0 to pH-12). At low and high pH, it shows drastic changes in the secondary and tertiary structure of the protein. EMSA studies showed the specific binding of Rv0047c with its own 30-bp promoter region. To determine the effect of complex formation on the secondary structure of Rv0047c, we examined the CD spectra of the complex of Rv0047c with promoter DNA of rv0047. The functional role of Rv0047c was characterized by over-expressing the Rv0047c gene under the control of hsp60 promoter in Mycobacterium tuberculosis H37Rv. We have predicted the three-dimensional structure of Rv0047c using the Swiss Model and Modeller, with validity checked by the Ramachandra plot. We did molecular docking of Rv0047c with dnaA, through PatchDock following refinement through FireDock. Through this, it is possible to easily identify the binding hot-stop of the receptor molecule with that of the ligand, the nature of the interface itself, and the conformational change undergone by the protein pattern. We are using X-crystallography to unravel the structure of Rv0047c. Overall the studies show that Rv0047c may have transcription regulation along with providing an insight into the activity of Rv0047c in the pH range of subcellular environment and helps to understand the protein-protein interaction, a novel target to kill dormant bacteria and potential strategy for tuberculosis control.

Keywords: mycobacterium tuberculosis, phenolic acid decarboxylase repressor, Rv0047c, Circular dichroism, fluorescence spectroscopy, docking, protein-protein interaction

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489 Binding Ability of Carbazolylphenyl Dendrimers with Zinc (II) Tetraphenylporphyrin Core towards Cryptands

Authors: Galina Mamardashvili, Nugzar Mamardashvili, Win Dehaen

Abstract:

The processes of complexation of the Zn-tetraarylporphyrins with eight 4-(4-(3,6-bis(t-butyl)carbazol-9-yl-phenyl)-1,2,3-triazole (ZnP1) and eight 4-(4-(3,6-di-tert-butyl-9-H-carbazol-9-yl)phenoxy)methyl)-2,4,6-trimethylphenyl (ZnP2)with the 1,10-diaza-4,7,13,18tetraoxabicyclo[8.5.5]eicosane (L1),1,10-diaza-4,7,13,16,21,24-hexaoxabicyclo[8.8.8]hexacosane (L2)and 1,10-diaza-5,6,14,15-dibenzo-4,7,13,16,21,24 hexaoxabicyclo[8.8.8] hexacosane (L3) were investigated by the method of spectrophotometric titration and 1H NMR-spectroscopy. We determined the structures of the host-guest complexes, and their stability constants in toluene were calculated. It was found out that the ZnP1 interacts with the guest molecules L1, L2 with the formation of stable "nest" type complexes and does not form similar complexes with the L3 (presumably due to the fact that the L3 does not match the size of the porphyrin ZnP(1) cavity). On the other hand, the porphyrin ZnP2 binds all of the ligands L1-L3, however complexes thus formed are less stable than complexes ZnP1-L1, ZnP1-L2. In the report, we will also discuss the influence of the alkali cations additives on the stability of the complexes between the porphyrin ZnP1, ZnP2 hosts and guest molecules of the ligands L1-L3.

Keywords: porphyrin, cryptand, cation, complex guest-host

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488 Screening Active Components in YPFS for Regulating Initiative Key Factors in Allergic Inflammation

Authors: Dandan Shen, Hui-zhu Wang, Xi Yu, LiLi Gui, Xiao Wei, Xiao-yan Jiang, Da-wei Wang, Min Hong

Abstract:

Yu-ping-feng-san (YPFS) is a clinical medicine for asthma and other allergic diseases, but the mechanism of YPFS on relapse of allergy is unclear. Currently, people come to realize the epithelial cells(EC) play a key role in stimulating and regulating local immune response. The study of thymic stromal lymphopoietin(TSLP derived from EC provides an important evidence that the EC can regulate immune response to stimulate allergic response. In this study, we observed the effect of YPFS on TSLP in vivo and in vitro. We established a method by using bronchial epithelial cells (16HBE) for screening potential bioactive components by HPLC-MS in YPFS and then analyzed the components in serum containing YPFS by UPLC-MS. The results showed that YPFS could decrease TSLP protein level in OVA-sensitized mice and 16HBE cells. Five components combing with the 16HBE cells were both detected in the serum.

Keywords: TSLP, bronchial epithelial cells, cell-binding, drug-containing serum

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487 Synthesis of Quinazoline Derivatives as Selective Inhibitors of Cyclooxygenase-1 Enzyme

Authors: Marcela Dvorakova, Lenka Langhansova, Premysl Landa

Abstract:

A series of quinazoline derivatives bearing aromatic rings in 2- and 4-positions were prepared and tested for their biological activity. Firstly, the compounds were evaluated for their potential to inhibit various kinases, such as autophagy activating kinase ULK1, 3-Phosphoinositide-dependent kinase 1, and TANK-binding kinase 1. None of the compounds displayed any activity on these kinases. Secondly, the compounds were tested for their anti-inflammatory activity expressed as cyclooxygenase (COX) isoforms and 5-lipoxygenase (5-LOX) inhibition. Three of the compounds showed significant selectivity towards COX-1 isoform (COX-2/COX-1 SI = 20-30). They inhibited COX-1 in a single-digit µM range. There was also one compound that exhibited inhibitory activity towards all three tested enzymes in µM range (IC50COX-1 = 1.9 µM; IC50COX-2 and 5-LOX = 10.1µM. COX-1 inhibition was until recently considered undesirable due to COX-1 constitutive expression in most cell types and tissues. Thus, there are not many compounds known with selective COX-1 activity. However, it is now believed that COX-1 plays an important role in the pathophysiology of several acute and chronic disorders, including cancer or neurodegenerative diseases. Thus, the discovery of effective COX-1 selective inhibitors is desirable and important.

Keywords: cyclooxygenases, kinases, lipoxygenases, quinazolines

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486 Sustainable Rehabilation of Ancient Structure

Authors: Ram Narayan Khare, Aradhna Shrivastava, Adhyatma Khare

Abstract:

This paper focuses on the damage that has been occurred in the Ancient structures due to various factors such as rainfall, climate, insects, lifespan and also most important lack of technologies in the era of its construction. The structure is of lime surkhi masonry and is made a century ago. It has crossed its durability but is of historical importance for the area, that is the reason why it needs utmost importance for its Rehabilitation. The paper deals with the damage that has been occurred in the structure and how to repair and renovate the same keeping in mind that the material deviation could not take place because it shows how in ancient era structures are made of. The building has used lime surkhi mortar along with wood apple as fibrous material for providing adhesiveness in masonry binding. The paper helps in sustainable retrofitting of the structure without changing the integrity of the structure. This helps in maintaining the originality of structure in present era and also help in providing information to the upcoming generation how ancient civil construction has been carried out that withstand even more than a century.

Keywords: Lime Surkhi masonry, rehabilitation, sustainable development, historical building

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485 The White Stork (Ciconia ciconia) in the Wetlands of North East of Algeria

Authors: Aicha Beya Mammeria, Idir Bitam

Abstract:

Our study focuses on the distribution of the white stork "Ciconia ciconia L. 1758" in the wetlands of El Tarf (North eastern of Algeria): recognized by its remarkable number of breeding pairs, monitoring of nesting, using a GPS has been performed in an attempt to explain the functioning of populations and population strategies for an overall design of its distribution, which has not so far been investigated in this region. Between 2012 and 2013, the number of breeding pairs has increased considerably from 174 in 1996 to 475 in 2007 and 968 in 2013. It should be noted that in the distribution of breeding pairs between 1996 and 2011, there is a significant development since the density of nests increased from 25.22 in 1996 to 84.16 couples/100 km² in 2013. More endemic bread apears in the region, this fluctuation is related to climatic change and changing season. Changes related to local climatic conditions might induce binding conditions for the development of this species.

Keywords: white stork, Ciconia ciconia, wetland El Tarf, northeast Algeria, climatic changing, density

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484 Biosynthesis of Titanium Dioxide Nanoparticles and Their Antibacterial Property

Authors: Prachi Singh

Abstract:

This paper presents a low-cost, eco-friendly and reproducible microbe mediated biosynthesis of TiO2 nanoparticles. TiO2 nanoparticles synthesized using the bacterium, Bacillus subtilis, from titanium as a precursor, were confirmed by TEM analysis. The morphological characteristics state spherical shape, with the size of individual or aggregate nanoparticles, around 30-40 nm. Microbial resistance represents a challenge for the scientific community to develop new bioactive compounds. Here, the antibacterial effect of TiO2 nanoparticles on Escherichia coli was investigated, which was confirmed by CFU (Colony-forming unit). Further, growth curve study of E. coli Hb101 in the presence and absence of TiO2 nanoparticles was done. Optical density decrease was observed with the increase in the concentration of TiO2. It could be attributed to the inactivation of cellular enzymes and DNA by binding to electron-donating groups such as carboxylates, amides, indoles, hydroxyls, thiols, etc. which cause little pores in bacterial cell walls, leading to increased permeability and cell death. This justifies that TiO2 nanoparticles have efficient antibacterial effect and have potential to be used as an antibacterial agent for different purposes.

Keywords: antibacterial effect, CFU, Escherichia coli Hb101, growth curve, TEM, TiO2 nanoparticle, Toxicity, UV-Vis

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483 Synthesis, Characterization and Application of Undoped and Fe Doped TiO₂ (Ti₁₋ₓFeₓO₂; X=0.01, 0.02, 0.03) Nanoparticles

Authors: Sudhakar Saroj, Satya Vir Singh

Abstract:

Undoped and Fe doped TiO₂, Ti₁₋ₓFeₓO₂ (x=0.00, 0.01, 0.03, 0.05, 0.07 and 0.09) have been synthesized by solution combustion method using Titanium (IV) oxide as a precursor, and also were characterized by XRD, DRS, FTIR, XPS, SEM, and EDX. The formation of anatase phase of undoped and Fe TiO₂ nanoparticles were confirmed by XRD, and the average crystallite size was determined by Debye-Scherer's equation. The DRS analysis indicates the shifting of light absorbance in visible region from UV region with increasing the doping concentration in TiO₂. The vibrational band of the Ti-O lattice was confirmed by the FT-IR spectrum. The XPS results confirm the presence of elements of titanium, oxygen and iron in the synthesized samples and determine the binding energy of elements. SEM image of the above-synthesized nanoparticles showed the spherical shape of nanoparticles. The purities of the synthesized nanoparticles were confirmed by EDX analysis. The photocatalytic activities of the synthesized nanoparticles were tested by studying the degradation of dye (Direct Blue 199) in the photocatalytic reactor. The Ti₀.₉₇Fe₀.₀₃O₂ photocatalyst shows highest photodegradation activity among all the synthesized undoped and Fe doped TiO₂ photocatalyst.

Keywords: direct blue 199, nanoparticles, TiO₂, photodegradation

Procedia PDF Downloads 236