Search results for: nucleic acid binding
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3975

Search results for: nucleic acid binding

2805 Study on the Neurotransmitters and Digestion of Amino Acids Affecting Psychological Chemical Imbalance

Authors: Yoonah Lee, Richard Kyung

Abstract:

With technological advances in the computational biomedical field, the ability to measure neurotransmitters’ chemical imbalances that affect depression and anxiety has been established. By comparing the thermodynamics stability of amino acid supplements, such as glutamine, tyrosine, phe-nylalanine, and methionine, this research analyzes mood-regulating neurotransmitters, amino acid supplements, and antipsychotic substances (ie. Reserpine molecule and CRF complexes) in relation to depression and anxiety and suggests alternative complexes that are low in energy to act as more efficient treatments for mood disorders. To determine a molecule’s thermodynamic stability, this research examines the molecular energy using Avogadro, a software for building virtual molecules and calculating optimized geometry using GAFF (General Amber Force Field) and UFF (Universal Force Field). The molecules, built using Avogadro, is analyzed using their theoretical values and atomic properties.

Keywords: amino acids, anxiety, depression, neurotransmitters

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2804 Molecular Characterization of Major Isolated Organism Involved in Bovine Subclinical Mastitis

Authors: H. K. Ratre, M. Roy, S. Roy, M. S. Parmar, V. Bhagat

Abstract:

Mastitis is a common problem of dairy industries. Reduction in milk production and an irreparable damage to the udder associated with the disease are common causes of culling of dairy cows. Milk from infected animals is not suitable for drinking and for making different milk products. So, it has a major economic importance in dairy cattle. The aims of this study were to investigate the bacteriological panorama in milk from udder quarters with subclinical mastitis and to carried out for the molecular characterization of the major isolated organisms, from subclinical mastitis-affected cows in and around Durg and Rajnandgaon district of Chhattisgarh. Isolation and identification of bacteria from the milk samples of subclinical mastitis-affected cows were done by standard and routine culture procedures. A total of 78 isolates were obtained from cows and among the various bacteria isolated, Staphylococcus spp. occupied prime position with occurrence rate of 51.282%. However, other bacteria isolated includeStreptococcus spp. (20.512%), Micrococcus spp. (14.102%), E. coli (8.974%), Klebsiela spp. (2.564%), Salmonella spp. (1.282%) and Proteus spp. (1.282%). Staphylococcus spp. was isolated as the major causative agent of subclinical mastitis in the studied area. Molecular characterization of Staphylococus aureusisolates was done for genetic expression of the virulence genes like ‘nuc’ encoding thermonucleaseexoenzyme, coa and spa by PCR amplification of the respective genes in 25 Staphylococcus isolates. In the present study, 15 isolates (77.27%) out of 20 coagulase positive isolates were found to be genotypically positive for ‘nuc’ where as 20 isolates (52.63%) out of 38 CNS expressed the presence of the same virulence gene. In the present study, three Staphylococcus isolates were found to be genotypically positive for coa gene. The Amplification of the coa gene yielded two different products of 627, 710 bp. The amplification of the gene segment encoding the IgG binding region of protein A (spa) revealed a size of 220 and 253bp in twostaphylococcus isolates. The X-region binding of the spa gene produced an amplicon of 315 bp in one Staphylococcal isolates. Staphylococcus aureus was found to be major isolate (51.28%) responsible for causing subclinical mastitis in cows which also showed expression of virulence genesnuc, coa and spa.

Keywords: mastitis, bacteria, characterization, expression, gene

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2803 Design and Development of Bioactive a-Hydroxy Carboxylate Group Modified MnFe₂O₄ Nanoparticle: Comparative Fluorescence Study, Magnetism and DNA Nuclease Activity

Authors: Indranil Chakraborty, Kalyan Mandal

Abstract:

Three new α-hydroxy carboxylate group functionalized MnFe₂O₄ nanoparticles (NPs) have been developed to explore the microscopic origin of ligand modified fluorescence and magnetic properties of nearly monodispersed MnFe₂O₄ NPs. The surface functionalization has been carried out with three small organic ligands (tartrate, malate, and citrate) having different number of α-hydroxy carboxylate functional group along with steric effect. Detailed study unveils that α-hydroxy carboxylate moiety of the ligands plays key role to generate intrinsic fluorescence in functionalized MnFe₂O₄ NPs through the activation of ligand to metal charge transfer transitions, associated with ligand-Mn²⁺/Fe³⁺ interactions along with d-d transition corresponding to d-orbital energy level splitting of Fe³⁺ ions on NP surface. Further, MnFe₂O₄ NPs show a maximum 140.88% increase in coercivity and 97.95% decrease in magnetization compared to its bare one upon functionalization. The ligands that induce smallest crystal field splitting of d-orbital energy level of transition metal ions are found to result in strongest ferromagnetic activation of the NPs. Finally, our developed tartrate functionalized MnFe₂O₄ (T-MnFe₂O₄) NPs have been utilized for studying DNA binding interaction and nuclease activity for stimulating their beneficial activities toward diverse biomedical applications. The spectroscopic measurements indicate that T-MnFe₂O₄ NPs bind calf thymus DNA by intercalative mode. The ability of T-MnFe₂O₄ NPs to induce DNA cleavage was studied by gel electrophoresis technique where the complex is found to promote the cleavage of pBR322 plasmid DNA from the super coiled form I to linear coiled form II and nicked coiled form III with good efficiency. This may be taken into account for designing new biomolecular detection agents and anti-cancer drug which can open up a new door toward diverse non-invasive biomedical applications.

Keywords: MnFe₂O₄ nanoparticle, α-hydroxy carboxylic acid, comparative fluorescence, magnetism study, DNA interaction, nuclease activity

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2802 Enhancement Effect of Compound 4-Hydroxybenzoic Acid from Petung Bamboo (Dendrocalamus Asper) Shoots on α1β2γ2S of GABA (A) Receptor Expressed in Xenopus laevis Oocytes- Preliminary Study on Its Anti-Epileptic Potential

Authors: Muhammad Bilal, Amelia Jane Llyod, Habsah Mohamad, Jia Hui Wong, Abdul Aziz Mohamed Yusoff, Jafri Malin Abdullah, Jingli Zhang

Abstract:

Epilepsy is one of the major brain afflictions occurs with uncontrolled excitation of cortex; disturbed 50 million of world’s population. About 25 percent of patients subjected to adverse effects from antiepileptic drugs (AEDs) such as depression, nausea, tremors, gastrointestinal symptoms, osteoporosis, dizziness, weight change, drowsiness, fatigue are commonly observed indications; therefore, new drugs are required to cure epilepsy. GABA is principle inhibitory neurotransmitter, control excitation of the brain. Mutation or dysfunction of GABA receptor is one of the primary causes of epilepsy, which is confirmed from many acquired models of epilepsy like traumatic brain injury, kindling, and status epilepticus models of epilepsy. GABA receptor has 3 distinct types such as GABA (A), GABA (B), GABA(C).GABA (A) receptor has 20 different subunits, α1β2γ2 subunits composition of GABA (A) receptor is the most used combination of subunits for screening of compounds against epilepsy. We expressed α1β2γ2s subunits of GABA (A) Receptor in Xenopus leavis oocytes and examined the enhancement potential of 4-Hydroxybenzoic acid compound on GABA (A) receptor via two-electrode voltage clamp current recording technique. Bamboo shoots are the young, tender offspring of bamboo, which are usually harvested after a cultivating period of 2 weeks. Proteins, acids, fat, starch, carbohydrate, fatty acid, vitamin, dietary fiber, and minerals are the major constituent found systematically in bamboo shoots. These shoots reported to have anticancer, antiviral, antibacterial activity, also possess antioxidant properties due to the presence of phenolic compounds. Student t-test analysis suggested that 4- hydroxybenzoic acid positively allosteric GABA (A) receptor, increased normalized current amplitude to 1.0304±0.0464(p value 0.032) compared with vehicle. 4-Hydrobenzoic acid, a compound from Dendrocalamus Asper bamboo shoot gives new insights for future studies on bamboo shoots with motivation for extraction of more compounds to investigate their effects on human and rodents against epilepsy, insomnia, and anxiety.

Keywords: α1β2γ2S, antiepileptic, bamboo shoots, epilepsy GABA (A) receptor, two-microelectrode voltage clamp, xenopus laevis oocytes

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2801 Evaluation of Antagonistic and Aggregation Property of Probiotic Lactic Acid Bacteria Isolated from Bovine Milk

Authors: Alazar Nebyou, Sujata Pandit

Abstract:

Lactic acid bacteria (LAB) are essential ingredients in probiotic foods, intestinal microflora, and dairy products that are capable of coping up with harsh gastrointestinal tract conditions and are available in a variety of environments. The objective of this study is to evaluate the probiotic property of LAB isolated from bovine milk. Milk samples were collected from local dairy farms. Samples were obtained using sterile test tubes and transported to a laboratory in the icebox for further biochemical characterization. Preliminary physiological and biochemical identification of LAB isolates was conducted by growing on MRS agar after ten-fold serial dilution. Seven of the best isolates were selected for the evaluation of the probiotic property. The LAB isolates were checked for resistance to antibiotics and their antimicrobial activity by disc diffusion assay and agar well diffusion assay respectively. Bile salt hydrolase activity of isolates was studied by growing isolates in a BSH medium with bile salt. Cell surface property of isolates was assayed by studying their autoaggregation and coaggregation percentage with S. aerues. All isolates were found BSH positive. In addition, BCM2 and BGM1 were susceptible to all antibiotic disks except BBM1 which was resistant to all antibiotic disks. BCM1 and BGM1 had the highest autoaggregation and coaggregation potential respectively. Since all LAB isolates showed gastrointestinal tolerance and good cell surface property they could be considered as good potential probiotic candidates for treatment and probiotic starter culture preparation.

Keywords: probiotic, aggregation, lactic acid bacteria, antimicrobial activity

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2800 Full-Face Hyaluronic Acid Implants Assisted by Artificial Intelligence-Generated Post-treatment 3D Models

Authors: Ciro Cursio, Pio Luigi Cursio, Giulia Cursio, Isabella Chiardi, Luigi Cursio

Abstract:

Introduction: Full-face aesthetic treatments often present a difficult task: since different patients possess different anatomical and tissue characteristics, there is no guarantee that the same treatment will have the same effect on multiple patients; additionally, full-face rejuvenation and beautification treatments require not only a high degree of technical skill but also the ability to choose the right product for each area and a keen artistic eye. Method: We present an artificial intelligence-based algorithm that can generate realistic post-treatment 3D models based on the patient’s requests together with the doctor’s input. These 3-dimensional predictions can be used by the practitioner for two purposes: firstly, they help ensure that the patient and the doctor are completely aligned on the expectations of the treatment; secondly, the doctor can use them as a visual guide, obtaining a natural result that would normally stem from the practitioner's artistic skills. To this end, the algorithm is able to predict injection zones, the type and quantity of hyaluronic acid, the injection depth, and the technique to use. Results: Our innovation consists in providing an objective visual representation of the patient that is helpful in the patient-doctor dialogue. The patient, based on this information, can express her desire to undergo a specific treatment or make changes to the therapeutic plan. In short, the patient becomes an active agent in the choices made before the treatment. Conclusion: We believe that this algorithm will reveal itself as a useful tool in the pre-treatment decision-making process to prevent both the patient and the doctor from making a leap into the dark.

Keywords: hyaluronic acid, fillers, full face, artificial intelligence, 3D

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2799 Enhanced Visible-Light Photocatalytic Activity of TiO2 Doped in Degradation of Acid Dye

Authors: B. Benalioua, I. Benyamina, M. Mansour, A. Bentouami, B. Boury

Abstract:

The objective of this study is based on the synthesis of a new photocatalyst based on TiO2 and its application in the photo-degradation of an acid dye under the visible light. The material obtained was characterized by XRD, BET and UV- vis DRS. The photocatalytic efficiency of the Zn -Fe TiO2 treated at 500°C was tested on the Indigo Carmine under the irradiation of visible light and compared with that of the commercial titanium oxide TiO2-P25 (Degussa). The XRD characterization of the material Zn-Fe-TiO2 (500°C) revealed the presence of the anatase phase and the absence of the Rutile phase in comparison of the TiO2 P25 diffractogram. Characterization by UV-visible diffuse reflection material showed that the Fe-Zn-TiO2 exhibits redshift (move visible) relative to commercial titanium oxide TiO2-P25, this property promises a photocatalytic activity of Zn -Fe- TiO2 under visible light. Indeed, the efficiency of photocatalytic Fe-Zn-TiO2 as a visible light is shown by a complete discoloration of indigo carmine solution of 16 mg/L after 40 minutes, whereas with the P25-TiO2 discoloration is achieved after 90 minutes.

Keywords: POA, heterogeneous photocatalysis, TiO2, doping

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2798 Development and Optimization of Colon Targeted Drug Delivery System of Ayurvedic Churna Formulation Using Eudragit L100 and Ethyl Cellulose as Coating Material

Authors: Anil Bhandari, Imran Khan Pathan, Peeyush K. Sharma, Rakesh K. Patel, Suresh Purohit

Abstract:

The purpose of this study was to prepare time and pH dependent release tablets of Ayurvedic Churna formulation and evaluate their advantages as colon targeted drug delivery system. The Vidangadi Churna was selected for this study which contains Embelin and Gallic acid. Embelin is used in Helminthiasis as therapeutic agent. Embelin is insoluble in water and unstable in gastric environment so it was formulated in time and pH dependent tablets coated with combination of two polymers Eudragit L100 and ethyl cellulose. The 150mg of core tablet of dried extract and lactose were prepared by wet granulation method. The compression coating was used in the polymer concentration of 150mg for both the layer as upper and lower coating tablet was investigated. The results showed that no release was found in 0.1 N HCl and pH 6.8 phosphate buffers for initial 5 hours and about 98.97% of the drug was released in pH 7.4 phosphate buffer in total 17 hours. The in vitro release profiles of drug from the formulation could be best expressed first order kinetics as highest linearity (r2= 0.9943). The results of the present study have demonstrated that the time and pH dependent tablets system is a promising vehicle for preventing rapid hydrolysis in gastric environment and improving oral bioavailability of Embelin and Gallic acid for treatment of Helminthiasis.

Keywords: embelin, gallic acid, Vidangadi Churna, colon targeted drug delivery

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2797 Preparation of Allyl BODIPY for the Click Reaction with Thioglycolic Acid

Authors: Chrislaura Carmo, Luca Deiana, Mafalda Laranjo, Abilio Sobral, Armando Cordova

Abstract:

Photodynamic therapy (PDT) is currently used for the treatment of malignancies and premalignant tumors. It is based on the capture of a photosensitizing molecule (PS) which, when excited by light at a certain wavelength, reacts with oxygen and generates oxidizing species (radicals, singlet oxygen, triplet species) in target tissues, leading to cell death. BODIPY (4,4-difluoro-4-bora-3a,4a-diaza-s-indaceno) derivatives are emerging as important candidates for photosensitizer in photodynamic therapy of cancer cells due to their high triplet quantum yield. Today these dyes are relevant molecules in photovoltaic materials and fluorescent sensors. In this study, it will be demonstrated the possibility that BODIPY can be covalently linked to thioglycolic acid through the click reaction. Thiol−ene click chemistry has become a powerful synthesis method in materials science and surface modification. The design of biobased allyl-terminated precursors with high renewable carbon content for the construction of the thiol-ene polymer networks is essential for sustainable development and green chemistry. The work aims to synthesize the BODIPY (10-(4-(allyloxy) phenyl)-2,8-diethyl-5,5-difluoro-1,3,7,9-tetramethyl-5H-dipyrrolo[1,2-c:2',1'-f] [1,3,2] diazaborinin-4-ium-5-uide) and to click reaction with Thioglycolic acid. BODIPY was synthesized by the condensation reaction between aldehyde and pyrrole in dichloromethane, followed by in situ complexation with BF3·OEt2 in the presence of the base. Then it was functionalized with allyl bromide to achieve the double bond and thus be able to carry out the click reaction. The thiol−ene click was performed using DMPA (2,2-Dimethoxy-2-phenylacetophenone) as a photo-initiator in the presence of UV light (320–500 nm) in DMF at room temperature for 24 hours. Compounds were characterized by standard analytical techniques, including UV-Vis Spectroscopy, 1H, 13C, 19F NMR and mass spectroscopy. The results of this study will be important to link BODIPY to polymers through the thiol group offering a diversity of applications and functionalization. This new molecule can be tested as third-generation photosensitizers, in which the dye is targeted by antibodies or nanocarriers by cells, mainly in cancer cells, PDT and Photodynamic Antimicrobial Chemotherapy (PACT). According to our studies, it was possible to visualize a click reaction between allyl BODIPY and thioglycolic acid. Our team will also test the reaction with other thiol groups for comparison. Further, we will do the click reaction of BODIPY with a natural polymer linked with a thiol group. The results of the above compounds will be tested in PDT assays on various lung cancer cell lines.

Keywords: bodipy, click reaction, thioglycolic acid, allyl, thiol-ene click

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2796 Composite Electrospun Aligned PLGA/Curcumin/Heparin Nanofibrous Membranes for Wound Dressing Application

Authors: Jyh-Ping Chen, Yu-Tin Lai

Abstract:

Wound healing is a complicated process involving overlapping hemostasis, inflammation, proliferation, and maturation phases. Ideal wound dressings can replace native skin functions in full thickness skin wounds through faster healing rate and also by reducing scar formation. Poly(lactic-co-glycolic acid) (PLGA) is an U.S. FDA approved biodegradable polymer to be used as ideal wound dressing material. Several in vitro and in vivo studies have demonstrated the effectiveness of curcumin in decreasing the release of inflammatory cytokines, inhibiting enzymes associated with inflammations, and scavenging free radicals that are the major cause of inflammation during wound healing. Heparin has binding affinities to various growth factors. With the unique and beneficial features offered by those molecules toward the complex process of wound healing, we postulate a composite wound dressing constructed from PLGA, curcumin and heparin would be a good candidate to accelerate scarless wound healing. In this work, we use electrospinning to prepare curcumin-loaded aligned PLGA nanofibrous membranes (PC NFMs). PC NFMs were further subject to oxygen plasma modification and surfaced-grafted with heparin through carbodiimide-mediated covalent bond formation to prepare curcumin-loaded PLGA-g-heparin (PCH) NFMs. The nanofibrous membranes could act as three-dimensional scaffolds to attract fibroblast migration, reduce inflammation, and increase wound-healing related growth factors concentrations at wound sites. From scanning electron microscopy analysis, the nanofibers in each NFM are with diameters ranging from 456 to 479 nm and with alignment angles within  0.5°. The NFMs show high tensile strength and good water absorptivity and provide suitable pore size for nutrients/wastes transport. Exposure of human dermal fibroblasts to the extraction medium of PC or PCH NFM showed significant protective effects against hydrogen peroxide than PLGA NFM. In vitro wound healing assays also showed that the extraction medium of PCH NFM showed significantly better migration ability toward fibroblasts than PC NFM, which is further better than PLGA NFM. The in vivo healing efficiency of the NFMs was further evaluated by a full thickness excisional wound healing diabetic rat model. After 14 days, PCH NFMs exhibits 86% wound closure rate, which is significantly different from other groups (79% for PC and 73% for PLGA NFM). Real-time PCR analysis indicated PC and PCH NFMs down regulated anti-oxidative enzymes like glutathione peroxidase (GPx) and superoxide dismutase (SOD), which are well-known transcription factors involved in cellular inflammatory responses to stimuli. From histology, the wound area treated with PCH NFMs showed more vascular lumen formation from immunohistochemistry of α-smooth muscle actin. The wound site also had more collagen type III (65.8%) expression and less collagen type I (3.5%) expression, indicating scar-less wound healing. From Western blot analysis, the PCH NFM showed good affinity toward growth factors from increased concentration of transforming growth factor-β (TGF-β) and fibroblast growth factor-2 (FGF-2) at the wound site to accelerate wound healing. From the results, we suggest PCH NFM as a promising candidate for wound dressing applications.

Keywords: Curcumin, heparin, nanofibrous membrane, poly(lactic-co-glycolic acid) (PLGA), wound dressing

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2795 Improvement of Antibacterial Activity for Ceftazidime by Partially Purified Tannase from Penicillium expansum

Authors: Sahira N. Muslim, Alaa N. Mohammed, Saba Saadoon Khazaal, Batool Kadham Salman, Israa M. S. AL-Kadmy, Sraa N. Muslim, Ahmed S. Dwaish, Sawsan Mohammed Kareem, Sarah N. Aziz, Ruaa A. Jasim

Abstract:

Tannase has wide applications in food, beverage, brewing, cosmetics and chemical industries and one of the major applications of tannase is the production of gallic acid. Gallic acid is used for manufacturing of trimethoprim. In the present study, a local fungal strain of Penicillium expansum A4 isolated from spoilt apple samples gave the highest production level of tannase. Tannase was partially purified with a recovery yield of 92.52% and 6.32 fold of purification by precipitation using ammonium sulfate at 50% saturation. Tannase led to increased antimicrobial activity of ceftazidime against Pseudomonas aeruginosa and S. aureus and had a synergism effect at low concentrations of ceftazidime, and thus, tannase may be a useful adjuvant agent for the treatment of many bacterial infections in combination with ceftazidime.

Keywords: ceftazidime, Penicillium expansum, tannase, antimicrobial activity

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2794 Co₂Fe LDH on Aromatic Acid Functionalized N Doped Graphene: Hybrid Electrocatalyst for Oxygen Evolution Reaction

Authors: Biswaranjan D. Mohapatra, Ipsha Hota, Swarna P. Mantry, Nibedita Behera, Kumar S. K. Varadwaj

Abstract:

Designing highly active and low-cost oxygen evolution (2H₂O → 4H⁺ + 4e⁻ + O₂) electrocatalyst is one of the most active areas of advanced energy research. Some precious metal-based electrocatalysts, such as IrO₂ and RuO₂, have shown excellent performance for oxygen evolution reaction (OER); however, they suffer from high-cost and low abundance which limits their applications. Recently, layered double hydroxides (LDHs), composed of layers of divalent and trivalent transition metal cations coordinated to hydroxide anions, have gathered attention as an alternative OER catalyst. However, LDHs are insulators and coupled with carbon materials for the electrocatalytic applications. Graphene covalently doped with nitrogen has been demonstrated to be an excellent electrocatalyst for energy conversion technologies such as; oxygen reduction reaction (ORR), oxygen evolution reaction (OER) & hydrogen evolution reaction (HER). However, they operate at high overpotentials, significantly above the thermodynamic standard potentials. Recently, we reported remarkably enhanced catalytic activity of benzoate or 1-pyrenebutyrate functionalized N-doped graphene towards the ORR in alkaline medium. The molecular and heteroatom co-doping on graphene is expected to tune the electronic structure of graphene. Therefore, an innovative catalyst architecture, in which LDHs are anchored on aromatic acid functionalized ‘N’ doped graphene may presumably boost the OER activity to a new benchmark. Herein, we report fabrication of Co₂Fe-LDH on aromatic acid (AA) functionalized ‘N’ doped reduced graphene oxide (NG) and studied their OER activities in alkaline medium. In the first step, a novel polyol method is applied for synthesis of AA functionalized NG, which is well dispersed in aqueous medium. In the second step, Co₂Fe LDH were grown on AA functionalized NG by co-precipitation method. The hybrid samples are abbreviated as Co₂Fe LDH/AA-NG, where AA is either Benzoic acid or 1, 3-Benzene dicarboxylic acid (BDA) or 1, 3, 5 Benzene tricarboxylic acid (BTA). The crystal structure and morphology of the samples were characterized by X-ray diffraction (XRD), scanning electron microscope (SEM) and transmission electron microscope (TEM). These studies confirmed the growth of layered single phase LDH. The electrocatalytic OER activity of these hybrid materials was investigated by rotating disc electrode (RDE) technique on a glassy carbon electrode. The linear sweep voltammetry (LSV) on these catalyst samples were taken at 1600rpm. We observed significant OER performance enhancement in terms of onset potential and current density on Co₂Fe LDH/BTA-NG hybrid, indicating the synergic effect. This exploration of molecular functionalization effect in doped graphene and LDH system may provide an excellent platform for innovative design of OER catalysts.

Keywords: π-π functionalization, layered double hydroxide, oxygen evolution reaction, reduced graphene oxide

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2793 Some Probiotic Traits of Lactobacillus Strains Isolated from Pollen

Authors: Hani Belhadj, Daoud Harzallah, Seddik Khennouf, Saliha Dahamna, Mouloud Ghadbane

Abstract:

In this study, Lactobacillus strains isolated from pollen were identified by means of phenotypic and genotypic methods, At pH 2, most strains proved to be acid resistants, with losses in cell viability ranging from 0.77 to 4.04 Log orders. In addition, at pH 3 all strains could grew and resist the acidic conditions, with losses in cell viability ranging from 0.40 to 3.61 Log orders. It seems that, 0.3% and 0.5% of bile salts does not affect greatly the survival of most strains, excluding Lactobacillus sp. BH1398. Survival ranged from 81.0±3.5 to 93.5±3.9%. In contrast, in the presence of 1.0% bile salts, survival of five strains was decreased by more than 50%. Lactobacillus fermentum BH1509 was considered the most tolerant strain (77.5% for 1% bile) followed by Lactobacillus plantarum BH1541 (59.9% for 1% bile). Furthermore, all strains were resistant to colistine, clindamycine, chloramphenicol, and ciprofloxacine, but most of the strains were susceptible to Peniciline, Oxacillin, Oxytetracyclin, and Amoxicillin. Functionally interesting Lactobacillus isolates may be used in the future as probiotic cultures for manufacturing fermented foods and as bioactive delivery systems.

Keywords: probiotics, lactobacillus, pollen, bile, acid tolerance

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2792 Zinc Borate Synthesis Using Hydrozincite and Boric Acid with Ultrasonic Method

Authors: D. S. Vardar, A. S. Kipcak, F. T. Senberber, E. M. Derun, S. Piskin, N. Tugrul

Abstract:

Zinc borate is an important inorganic hydrate borate material, which can be use as a flame retardant agent and corrosion resistance material. This compound can loss its structural water content at higher than 290°C. Due to thermal stability; Zinc Borate can be used as flame reterdant at high temperature process of plastic and gum. In this study, the ultrasonic reaction of zinc borates were studied using hydrozincite (Zn5(CO3)2•(OH)6) and boric acid (H3BO3) raw materials. Before the synthesis raw materials were characterized by X-Ray Diffraction (XRD) and Fourier Transform Infrared Spectroscopy (FT-IR). Ultrasonic method is a new application on the zinc borate synthesis. The synthesis parameters were set to 90°C reaction temperature and 55 minutes of reaction time, with 1:1, 1:2, 1:3, 1:4 and 1:5 molar ratio of starting materials (Zn5(CO3)2•(OH)6 : H3BO3). After the zinc borate synthesis, the products analyzed by XRD and FT-IR. As a result, optimum molar ratio of 1:5 (Zn5(CO3)2•(OH)6:H3BO3) is determined for the synthesis of zinc borates with ultrasonic method.

Keywords: borate, ultrasonic method, zinc borate, zinc borate synthesis

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2791 Some Quality Characteristics of Red Raspberry Jam

Authors: Cristina Damian, Eduard Malcek, Ana Leahu, Sorina Ropciuc, Andrei Lobiuc

Abstract:

Given the high rivalry nowadays, the food sector must offer the markets an attractive product, which at the same time has good quality and is safe from health aspects for the consumers. Known for their high content of antioxidant compounds, especially anthocyanins, which proven human health benefits, berries from the Rosaceae family plants have a significantly high level of phytochemicals: phenolic flavonoids, such as anthocyanins, ellagic acid (tannin), quercetin, gallic acid, cyanidin, pelargonidine, catechins, kaempferol and salicylic acid. Colour and bioactive compounds, such as vitamin C and anthocyanins, are important for the attractiveness of berries and their preserved products. The levels of bioactive compounds and sensory properties of the product as it reaches the consumer are dependent on raw material, i.e., berries used, processing and storage conditions. In this study, four varieties of raspberry jam were analyzed, 3 of them purchased commercially; they were purchased at reasonable prices, precisely to include as large a sample of the consumer population as possible. The fourth assortment was made at home according to the traditional recipe without the addition of sweeteners or preservatives. As for the homemade red raspberry jam, it had a sugar concentration of 64.9%, being the most appreciated of all assortments. The homemade raspberry jam was most appreciated due to the taste and aroma of the product. The SCHWARTAU assortment was chosen in second place by the participants in the study (sensory analysis). The quality/price ratio is also valid this time, finding that a high-quality product will have a higher purchase price. Thus, the study had the role of presenting the preferences of the sample participating in the study by age categories.

Keywords: red rasperry, jam, antioxidant, colour, sensory analysis

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2790 The Understanding of Biochemical and Molecular Analysis of Diabetic Rats Treated with Andrographis paniculata and Erythrina indica Methanol Extract

Authors: Chakrapani Pullagummi, Arun Jyothi Bheemagani, B. Chandra Sekhar Singh, Prem Kumar, A. Roja Rani

Abstract:

Diabetes mellitus describes a metabolic disorder of multiple aetiology characterized by chronic hyperglycaemia with disturbances of carbohydrate, fat and protein metabolism resulting from defects in insulin secretion and its action. The objective of present study was alloxan induced diabetes in S.D (Sprague Dawley) rats, treated with leaf extract of Andrographis paniculata and bark extract of Erythrina indica. Plant extract treated rats were analyzed biochemically and molecularly. on normal and diabetic rats. The changes in MDA (lipid peroxidation) and glucose (by GOD method) levels in blood of both normal and diabetic rat were analyzed. Diabetes induced rats were treated with methanolic extracts of Andrographis paniculata leaf and Erythrina indica bark which are of medicinal importance. Later after inducing diabetes the rats were treated with medicinal plant extracts, Andrographis paniculata leaf and Erythrina indica bark which are well known for their anti diabetic and antioxidative property in order to control the glucose and MDA levels. The blood plasma of diabetic and normal rats was analyzed for the levels of MDA (lipid peroxidation) and glucose levels. Results of this study suggested that the Andrographis paniculata leaf and Erythrina indica can be used as a potential natural antidiabetic agent for treating and postponing the appearance of complications that arise due to Diabetes. Molecular study deals with the analysis of binding mechanism of 2 selected natural compounds from Andrographis and Erythrina extracts against the novel target for type T2D namely PPAR-γ compared with Rosiglitazone (standard compound). The results revealed that most of the selected herbal lead compounds were effective targets against the receptors. These compounds showed favorable interactions with the amino acid residues thereby substantiating their proven efficacy as anti-diabetic compounds.

Keywords: andrographis paniculata, erythrina indica, alloxan, lipid peroxidation, blood glucose level, PPAR-γ

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2789 Risk Assessment and Haloacetic Acids Exposure in Drinking Water in Tunja, Colombia

Authors: Bibiana Matilde Bernal Gómez, Manuel Salvador Rodríguez Susa, Mildred Fernanda Lemus Perez

Abstract:

In chlorinated drinking water, Haloacetic acids have been identified and are classified as disinfection byproducts originating from reaction between natural organic matter and/or bromide ions in water sources. These byproducts can be generated through a variety of chemical and pharmaceutical processes. The term ‘Total Haloacetic Acids’ (THAAs) is used to describe the cumulative concentration of dichloroacetic acid, trichloroacetic acid, monochloroacetic acid, monobromoacetic acid, and dibromoacetic acid in water samples, which are usually measured to evaluate water quality. Chronic presence of these acids in drinking water has a risk of cancer in humans. The detection of THAAs for the first time in 15 municipalities of Boyacá was accomplished in 2023. Aim is to describe the correlation between the levels of THAAs and digestive cancer in Tunja, a city in Colombia with higher rates of digestive cancer and to compare the risk across 15 towns, taking into account factors such as water quality. A research project was conducted with the aim of comparing water sources based on the geographical features of the town, describing the disinfection process in 15 municipalities, and exploring physical properties such as water temperature and pH level. The project also involved a study of contact time based on habits documented through a survey, and a comparison of socioeconomic factors and lifestyle, in order to assess the personal risk of exposure. Data on the levels of THAAs were obtained after characterizing the water quality in urban sectors in eight months of 2022. This, based on the protocol described in the Stage 2 DBP of the United States Environmental Protection Agency (USEPA) from 2006, which takes into account the size of the population being supplied. A cancer risk assessment was conducted to evaluate the likelihood of an individual developing cancer due to exposure to pollutants THAAs. The assessment considered exposure methods like oral ingestion, skin absorption, and inhalation. The chronic daily intake (CDI) for these exposure routes was calculated using specific equations. The lifetime cancer risk (LCR) was then determined by adding the cancer risks from the three exposure routes for each HAA. The risk assessment process involved four phases: exposure assessment, toxicity evaluation, data gathering and analysis, and risk definition and management. The results conclude that there is a cumulative higher risk of digestive cancer due to THAAs exposure in drinking water.

Keywords: haloacetic acids, drinking water, water quality, cancer risk assessment

Procedia PDF Downloads 52
2788 Triggering Apoptosis to Uproot Breast Cancer: HPLC-MS/MS Profiling, in-vitro and in-silico Fascinating Results of Polyphenolics in Pomegranate Rind Extract

Authors: Alaa M. Badr Eldin, Mayar M. Shahen, Mohammed S. Sedeek, Marwa I. Ezzat, Sawsan M. ElSonbaty, Muhammed A. Saad, Manal S. Afifi, Omar M. Sabry

Abstract:

Using HPLC-MS/MS technique, 133 polyphenolic compounds were identified in the methanol extract of pomegranate rind (Punica granatum L.). In-vitro cytotoxic activity against breast cancer cell line MCF-7 was investigated, with an IC50 of 54 ug/ml. In-silico molecular docking using ellagic acid, gallagic acid, and Punicalagin as model compounds identified in pomegranate rind extract confirmed the intriguing anti-estrogenic action of the key polyphenolic components in pomegranate rind extract. Surprisingly, taxol showed low activity compared to pomegranate compounds as ERα antagonist and ERβ agonist. Pomegranate rind extract enhanced apoptosis of breast cancer cells through upregulation of the caspase-3 expression and downregulation of NF-κB transcription factor.

Keywords: HPLC-MS/MS, pomegranate rind, cytotoxicity, MCF-7, ER, caspase-3, NF-kB

Procedia PDF Downloads 111
2787 Effect of Chemical Modifier on the Properties of Polypropylene (PP) / Coconut Fiber (CF) in Automotive Application

Authors: K. Shahril, A. Nizam, M. Sabri, A. Siti Rohana, H. Salmah

Abstract:

Chemical modifier (Acrylic Acid) is used as filler treatment to improve mechanical properties and swelling behavior of polypropylene/coconut fiber (PP/CF) composites by creating more adherent bonding between CF filler and PP Matrix. Treated (with chemical modifier) and untreated (without chemical modifier) composites were prepared in the formulation of 10 wt%, 20 wt%, 30 wt%, and 40 wt%. The mechanical testing indicates that composite with 10 wt% of untreated composite has the optimum value of tensile strength, and the composite with chemical modifier shows the tensile strength was increased. By increasing of filler loading, elastic modulus was increased while the elongation at brake was decreased. Meanwhile, the swelling test discerned that the increase of filler loading increased the water absorption of composites and the presence of chemical modifier reduced the equilibrium water absorption percentage.

Keywords: coconut fiber, polypropylene, acid acrylic, ethanol, chemical modifier, composites

Procedia PDF Downloads 457
2786 Investigation in Gassy Ozone Influence on Flaxes Made from Biologically Activated Whole Wheat Grains Quality Parameters

Authors: Tatjana Rakcejeva, Jelena Zagorska, Elina Zvezdina

Abstract:

The aim of the current research was to investigate the gassy ozone effect on quality parameters of flaxes made form whole biologically activated wheat grains. The research was accomplished on in year 2012 harvested wheat grains variety ′Zentos′. Grains were washed, wetted; grain biological activation was performed in the climatic chamber up to 24 hours. After biological activation grains was compressed; than flaxes was dried in convective drier till constant moisture content 9±1%. For grain treatment gassy ozone concentration as 0.0002% and treatment time – 6 min was used. In the processed flaxes the content of A and G tocopherol decrease by 23% and by 9%; content of B2 and B6 vitamins – by 11% and by 10%; elaidic acid – by 46%, oleic acid – by 29%; arginine (by 80%), glutamine (by 74%), asparagine and serine (by 68%), valine (by 62%), cysteine (by 54%) and tyrosine (by 47%).

Keywords: gassy ozone, flaxes, biologically activated grains, quality parameters, treatment

Procedia PDF Downloads 233
2785 Effect of Anion and Amino Functional Group on Resin for Lipase Immobilization with Adsorption-Cross Linking Method

Authors: Heri Hermansyah, Annisa Kurnia, A. Vania Anisya, Adi Surjosatyo, Yopi Sunarya, Rita Arbianti, Tania Surya Utami

Abstract:

Lipase is one of biocatalyst which is applied commercially for the process in industries, such as bioenergy, food, and pharmaceutical industry. Nowadays, biocatalysts are preferred in industries because they work in mild condition, high specificity, and reduce energy consumption (high pressure and temperature). But, the usage of lipase for industry scale is limited by economic reason due to the high price of lipase and difficulty of the separation system. Immobilization of lipase is one of the solutions to maintain the activity of lipase and reduce separation system in the process. Therefore, we conduct a study about lipase immobilization with the adsorption-cross linking method using glutaraldehyde because this method produces high enzyme loading and stability. Lipase is immobilized on different kind of resin with the various functional group. Highest enzyme loading (76.69%) was achieved by lipase immobilized on anion macroporous which have anion functional group (OH). However, highest activity (24,69 U/g support) through olive oil emulsion method was achieved by lipase immobilized on anion macroporous-chitosan which have amino (NH2) and anion (OH-) functional group. In addition, it also success to produce biodiesel until reach yield 50,6% through interesterification reaction and after 4 cycles stable 63.9% relative with initial yield. While for Aspergillus, niger lipase immobilized on anion macroporous-kitosan have unit activity 22,84 U/g resin and yield biodiesel higher than commercial lipase (69,1%) and after 4 cycles stable reach 70.6% relative from initial yield. This shows that optimum functional group on support for immobilization with adsorption-cross linking is the support that contains amino (NH2) and anion (OH-) functional group because they can react with glutaraldehyde and binding with enzyme prevent desorption of lipase from support through binding lipase with a functional group on support.

Keywords: adsorption-cross linking, immobilization, lipase, resin

Procedia PDF Downloads 364
2784 Synthesis of Magnesium Borates from the Slurries of Magnesium Wastes by Microwave Energy

Authors: N. Tugrul, F. T. Senberber, A. S. Kipcak, E. Moroydor Derun, S. Piskin

Abstract:

In this research, it is aimed not only microwave synthesis of magnesium borates but also evaluation of magnesium wastes. Synthesis process can be described with the reaction of Mg wastes and boric acid using microwave energy. X-Ray Diffraction (XRD) and Fourier Transform Infrared Spectroscopy (FT-IR) were applied to synthesized minerals. According to XRD results, magnesium borate hydrate mixtures were obtained as mcallisterite (pdf# = 01-070-1902, Mg2(B6O7(OH)6)2.9(H2O)) at higher crystallinity properties was achieved at the mole ratio raw material 1:1. Also, other kinds of magnesium borate hydrates were obtained at lower crystallinity such as admontite (pdf # = 01-076-0540, MgO(B2O3)3.7(H2O)), inderite (pdf # = 01-072-2308, 2MgO.3B2O3.15(H2O)) and magnesium borate hydrates (pdf # = 01-076-0539, MgO(B2O3)3.6(H2O)). FT-IR spectrums indicated that minor changes were seen at the band values of characteristic stretching in each experiment. At the end of experiments it is seen that using microwave energy may contribute positive effects to design of synthesis process such as reducing reaction time and products at higher crystallinity.

Keywords: magnesium wastes, boric acid, magnesium borate, microwave energy

Procedia PDF Downloads 351
2783 Methodology for the Determination of Triterpenic Compounds in Apple Extracts

Authors: Mindaugas Liaudanskas, Darius Kviklys, Kristina Zymonė, Raimondas Raudonis, Jonas Viškelis, Norbertas Uselis, Pranas Viškelis, Valdimaras Janulis

Abstract:

Apples are among the most commonly consumed fruits in the world. Based on data from the year 2014, approximately 84.63 million tons of apples are grown per annum. Apples are widely used in food industry to produce various products and drinks (juice, wine, and cider); they are also used unprocessed. Apples in human diet are an important source of different groups of biological active compounds that can positively contribute to the prevention of various diseases. They are a source of various biologically active substances – especially vitamins, organic acids, micro- and macro-elements, pectins, and phenolic, triterpenic, and other compounds. Triterpenic compounds, which are characterized by versatile biological activity, are the biologically active compounds found in apples that are among the most promising and most significant for human health. A specific analytical procedure including sample preparation and High Performance Liquid Chromatography (HPLC) analysis was developed, optimized, and validated for the detection of triterpenic compounds in the samples of different apples, their peels, and flesh from widespread apple cultivars 'Aldas', 'Auksis', 'Connel Red', 'Ligol', 'Lodel', and 'Rajka' grown in Lithuanian climatic conditions. The conditions for triterpenic compound extraction were optimized: the solvent of the extraction was 100% (v/v) acetone, and the extraction was performed in an ultrasound bath for 10 min. Isocratic elution (the eluents ratio being 88% (solvent A) and 12% (solvent B)) for a rapid separation of triterpenic compounds was performed. The validation of the methodology was performed on the basis of the ICH recommendations. The following characteristics of validation were evaluated: the selectivity of the method (specificity), precision, the detection and quantitation limits of the analytes, and linearity. The obtained parameters values confirm suitability of methodology to perform analysis of triterpenic compounds. Using the optimised and validated HPLC technique, four triterpenic compounds were separated and identified, and their specificity was confirmed. These compounds were corosolic acid, betulinic acid, oleanolic acid, and ursolic acid. Ursolic acid was the dominant compound in all the tested apple samples. The detected amount of betulinic acid was the lowest of all the identified triterpenic compounds. The greatest amounts of triterpenic compounds were detected in whole apple and apple peel samples of the 'Lodel' cultivar, and thus apples and apple extracts of this cultivar are potentially valuable for use in medical practice, for the prevention of various diseases, for adjunct therapy, for the isolation of individual compounds with a specific biological effect, and for the development and production of dietary supplements and functional food enriched in biologically active compounds. Acknowledgements. This work was supported by a grant from the Research Council of Lithuania, project No. MIP-17-8.

Keywords: apples, HPLC, triterpenic compounds, validation

Procedia PDF Downloads 168
2782 Current Issues of Cross-Border Enforcement

Authors: Gábor Kocsmárik

Abstract:

The topic of this is coercive measures against assets in which the factor of the procedure contains a foreign element. We speak of cross-border enforcement if the debtor or the property requesting enforcement or subject to enforcement is not located in the bordering country. Given that the jurisdiction of a country cannot extend beyond its borders, the cooperation of nations and the mutual recognition of their decisions are necessary to eliminate this. In addition, it is essential to create framework rules that are binding and enforceable for each country participating in the convention. During the study, some conventions between countries that are still in force will be presented, which can serve as a starting point for dealing with existing problems.

Keywords: law, execution, civil procedure law, international

Procedia PDF Downloads 30
2781 Phenotypic and Molecular Heterogeneity Linked to the Magnesium Transporter CNNM2

Authors: Reham Khalaf-Nazzal, Imad Dweikat, Paula Gimenez, Iker Oyenarte, Alfonso Martinez-Cruz, Domonik Muller

Abstract:

Metal cation transport mediator (CNNM) gene family comprises 4 isoforms that are expressed in various human tissues. Structurally, CNNMs are complex proteins that contain an extracellular N-terminal domain preceding a DUF21 transmembrane domain, a ‘Bateman module’ and a C-terminal cNMP-binding domain. Mutations in CNNM2 cause familial dominant hypomagnesaemia. Growing evidence highlights the role of CNNM2 in neurodevelopment. Mutations in CNNM2 have been implicated in epilepsy, intellectual disability, schizophrenia, and others. In the present study, we aim to elucidate the function of CNNM2 in the developing brain. Thus, we present the genetic origin of symptoms in two family cohorts. In the first family, three siblings of a consanguineous Palestinian family in which parents are first cousins, and consanguinity ran over several generations, presented a varying degree of intellectual disability, cone-rod dystrophy, and autism spectrum disorder. Exome sequencing and segregation analysis revealed the presence of homozygous pathogenic mutation in the CNNM2 gene, the parents were heterozygous for that gene mutation. Magnesium blood levels were normal in the three children and their parents in several measurements. They had no symptoms of hypomagnesemia. The CNNM2 mutation in this family was found to locate in the CBS1 domain of the CNNM2 protein. The crystal structure of the mutated CNNM2 protein was not significantly different from the wild-type protein, and the binding of AMP or MgATP was not dramatically affected. This suggests that the CBS1 domain could be involved in pure neurodevelopmental functions independent of its magnesium-handling role, and this mutation could have affected a protein partner binding or other functions in this protein. In the second family, another autosomal dominant CNNM2 mutation was found to run in a large family with multiple individuals over three generations. All affected family members had hypomagnesemia and hypermagnesuria. Oral supplementation of magnesium did not increase the levels of magnesium in serum significantly. Some affected members of this family have defects in fine motor skills such as dyslexia and dyslalia. The detected mutation is located in the N-terminal part, which contains a signal peptide thought to be involved in the sorting and routing of the protein. In this project, we describe heterogenous clinical phenotypes related to CNNM2 mutations and protein functions. In the first family, and up to the authors’ knowledge, we report for the first time the involvement of CNNM2 in retinal photoreceptor development and function. In addition, we report the presence of a neurophenotype independent of magnesium status related to the CNNM2 protein mutation. Taking into account the different modes of inheritance and the different positions of the mutations within CNNM2 and its different structural and functional domains, it is likely that CNNM2 might be involved in a wide spectrum of neuropsychiatric comorbidities with considerable varying phenotypes.

Keywords: magnesium transport, autosomal recessive, autism, neurodevelopment, CBS domain

Procedia PDF Downloads 145
2780 Dendroremediation of a Defunct Lead Acid Battery Recycling Site

Authors: Alejandro Ruiz-Olivares, M. del Carmen González-Chávez, Rogelio Carrillo-González, Martha Reyes-Ramos, Javier Suárez Espinosa

Abstract:

Use of automobiles has increased and proportionally, the demand for batteries to impulse them. When the device is aged, all the battery materials are reused through lead acid battery recycling (LABR). Importation of used lead acid batteries in Mexico has increased in the last years since many recycling factories have been settled in the country. Inadequate disposal of lead-acid battery recycling (LABR) wastes left soil severely polluted with Pb, Cu, and salts (Na+, SO2− 4, PO3− 4). Soil organic amendments may contribute with essential nutrients and sequester (scavenger compounds) metals to allow plant establishment. The objective of this research was to revegetate a former lead-acid battery recycling site aided with organic amendments. Seven tree species (Acacia farnesiana, Casuarina equisetifolia, Cupressus lusitanica, Eucalyptus obliqua, Fraxinus excelsior, Prosopis laevigata and Pinus greggii) and two organic amendments (vermicompost and vermicompost + sawdust mixture) were tested for phytoremediation of a defunct LABR site. Plants were irrigated during the dry season. Monitoring of the soils was carried out during the experiment: Available metals, salts concentrations and their spatial pattern in soil were analyzed. Plant species and amendments were compared through analysis of covariance and longitudinal analysis. High concentrations of extractable (DTPA-TEA-CaCl₂) metals (up to 15,685 mg kg⁻¹ and 478 mg kg⁻¹ for Pb and Cu) and soluble salts (292 mg kg-1 and 23,578 mg kg-1 for PO3− 4and SO2− 4) were found in the soil after three and six months of setting up the experiment. Lead and Cu concentrations were depleted in the rhizosphere after amendments addition. Spatial pattern of PO3− 4, SO2− 4 and DTPA-extractable Pb and Cu changed slightly through time. In spite of extreme soil conditions the plant species planted: A. farnesiana, E. obliqua, C. equisetifolia and F. excelsior had 100% of survival. Available metals and salts differently affected each species. In addition, negative effect on growth due to Pb accumulated in shoots was observed only in C. lusitanica. Many specimens accumulated high concentrations of Pb ( > 1000 mg kg-1) in shoots. C. equisetifolia and C. lusitanica had the best rate of growth. Based on the results, all the evaluated species may be useful for revegetation of Pb-polluted soils. Besides their use in phytoremediation, some ecosystem services can be obtained from the woodland such as encourage wildlife, wood production, and carbon sequestration. Further research should be conducted to analyze these services.

Keywords: heavy metals, inadequate disposal, organic amendments, phytoremediation with trees

Procedia PDF Downloads 277
2779 Engineering of Reagentless Fluorescence Biosensors Based on Single-Chain Antibody Fragments

Authors: Christian Fercher, Jiaul Islam, Simon R. Corrie

Abstract:

Fluorescence-based immunodiagnostics are an emerging field in biosensor development and exhibit several advantages over traditional detection methods. While various affinity biosensors have been developed to generate a fluorescence signal upon sensing varying concentrations of analytes, reagentless, reversible, and continuous monitoring of complex biological samples remains challenging. Here, we aimed to genetically engineer biosensors based on single-chain antibody fragments (scFv) that are site-specifically labeled with environmentally sensitive fluorescent unnatural amino acids (UAA). A rational design approach resulted in quantifiable analyte-dependent changes in peak fluorescence emission wavelength and enabled antigen detection in vitro. Incorporation of a polarity indicator within the topological neighborhood of the antigen-binding interface generated a titratable wavelength blueshift with nanomolar detection limits. In order to ensure continuous analyte monitoring, scFv candidates with fast binding and dissociation kinetics were selected from a genetic library employing a high-throughput phage display and affinity screening approach. Initial rankings were further refined towards rapid dissociation kinetics using bio-layer interferometry (BLI) and surface plasmon resonance (SPR). The most promising candidates were expressed, purified to homogeneity, and tested for their potential to detect biomarkers in a continuous microfluidic-based assay. Variations of dissociation kinetics within an order of magnitude were achieved without compromising the specificity of the antibody fragments. This approach is generally applicable to numerous antibody/antigen combinations and currently awaits integration in a wide range of assay platforms for one-step protein quantification.

Keywords: antibody engineering, biosensor, phage display, unnatural amino acids

Procedia PDF Downloads 138
2778 Bioinformatic Design of a Non-toxic Modified Adjuvant from the Native A1 Structure of Cholera Toxin with Membrane Synthetic Peptide of Naegleria fowleri

Authors: Frida Carrillo Morales, Maria Maricela Carrasco Yépez, Saúl Rojas Hernández

Abstract:

Naegleria fowleri is the causative agent of primary amebic meningoencephalitis, this disease is acute and fulminant that affects humans. It has been reported that despite the existence of therapeutic options against this disease, its mortality rate is 97%. Therefore, the need arises to have vaccines that confer protection against this disease and, in addition to developing adjuvants to enhance the immune response. In this regard, in our work group, we obtained a peptide designed from the membrane protein MP2CL5 of Naegleria fowleri called Smp145 that was shown to be immunogenic; however, it would be of great importance to enhance its immunological response, being able to co-administer it with a non-toxic adjuvant. Therefore, the objective of this work was to carry out the bioinformatic design of a peptide of the Naegleria fowleri membrane protein MP2CL5 conjugated with a non-toxic modified adjuvant from the native A1 structure of Cholera Toxin. For which different bioinformatics tools were used to obtain a model with a modification in amino acid 61 of the A1 subunit of the CT (CTA1), to which the Smp145 peptide was added and both molecules were joined with a 13-glycine linker. As for the results obtained, the modification in CTA1 bound to the peptide produces a reduction in the toxicity of the molecule in in silico experiments, likewise, the prediction in the binding of Smp145 to the receptor of B cells suggests that the molecule is directed in specifically to the BCR receptor, decreasing its native enzymatic activity. The stereochemical evaluation showed that the generated model has a high number of adequately predicted residues. In the ERRAT test, the confidence with which it is possible to reject regions that exceed the error values was evaluated, in the generated model, a high score was obtained, which determines that the model has a good structural resolution. Therefore, the design of the conjugated peptide in this work will allow us to proceed with its chemical synthesis and subsequently be able to use it in the mouse meningitis protection model caused by N. fowleri.

Keywords: immunology, vaccines, pathogens, infectious disease

Procedia PDF Downloads 82
2777 Response of Canola Traits to Integrated Fertilization Systems

Authors: Khosro Mohammadi

Abstract:

In order to study the effect of different resources of farmyard manure, compost and biofertilizers on grain yield and quality of canola (Talaieh cultivar), an experiment was conducted at Kurdistan region. Experimental units were arranged in split-split plots design based on randomized complete blocks with three replications. Main plots consisted of two locations with difference in soil texture (L1): Agricultural Research Center of Sanandaj and (L2): Islamic Azad University of Sanandaj, as location levels. Also, five strategies for obtaining the base fertilizer requirement including (N1): farmyard manure; (N2): compost; (N3): chemical fertilizers; (N4): farm yard manure + compost and (N5): farm yard manure + compost + chemical fertilizers were considered in split plots. Four levels of biofertilizers were (B1): Bacillus lentus and Pseudomonas putida; (B2): Trichoderma harzianum; (B3): Bacillus lentus and Pseudomonas putida & Trichoderma harzianum; and (B4): control. Results showed that location, different resources of fertilizer and interactions of them have a significant effect on grain yield. The highest grain yield (4660 kg/ha) was obtained from treatment, that farmyard manure, compost and biofertilizers were co application in clay loam soil (Gerizeh station). Different methods of fertilization have a significant effect on leaf chlorophyll. Highest amount of chlorophyll (38 Spad) was obtained from co application of farmyard manure, chemical fertilizers and compost (N5 treatment). Location, basal fertilizers and biofertilizers have a significant effect on N, S and N/S of canola seed. Oil content was decreased in Gerizeh station, but oil yield had a significant increasing than Azad University station. Co application of compost and farmyard manure produced highest percent of oleic acid (61.5 %) and linoleic acid (22.9 %). Co application of compost and farmyard manure has a significant increase in oleic acid and linoleic acid. Finally, L1N5B3 treatment, that compost, farmyard manure and biofertilizers were co application in Gerizeh station in compare to other treatments, selected as a best treatment of experiment.

Keywords: soil texture, organic fertilizer, chemical fertilizer, oil, Canola

Procedia PDF Downloads 396
2776 Antimicrobial Action and Its Underlying Mechanism by Methanolic Seed Extract of Syzygium cumini on Bacillus subtilis

Authors: Alok Kumar Yadav, Saurabh Saraswat, Preeti Sirohi, Manjoo Rani, Sameer Srivastava, Manish Pratap Singh, Nand K. Singh

Abstract:

The development of antibiotic resistance in bacteria is increasing at an alarming rate, and this is considered as one of the most serious threats in the history of medicine, and an alternative solution should be derived so as to tackle this problem. In many countries, people use the medicinal plants for the treatment of various diseases as these are cheaper, easily available and least toxic. Syzygium cumini is used for the treatment of various kinds of diseases but their mechanism of action is not reported. The antimicrobial activity of Syzygium cumini was tested by the well diffusion assay and zone of inhibition was reported to be 20.06 mm as compared to control with MIC of 0.3 mg/ml. Genomic DNA fragmentation of Bacillus subtilis revealed apoptosis and FE-SEM indicate cell wall cracking on several intervals of time. Propidium iodide staining results showed that few bacterial cells were stained in the control and population of stained cells increase after exposing them for various period of time. Flow cytometric kinetic data analysis on the membrane permeabilization in bacterial cell showed the significant contribution of antimicrobial potential of the seed extract on antimicrobial-induced permeabilization. Two components of Syzygium cumini methanolic seed extract was found to be quite active against four enzymes like PDB ID- 1W5D, 4OX3, 3MFD and 5E2F which have a very crucial role in membrane synthesis in Bacillus subtilis by in silico analysis. Through in silico analysis, lupeol showed highest binding energy for macromolecule 1W5D and 4OX3 whereas stigmasterol showed the highest binding energy for macromolecule 3MFD and 5E2F respectively. It showed that methanolic seed extract of Syzygium cumini can be used for the inhibition of foodborne infections caused by Bacillus subtilis and also as an alternative of prevalent antibiotics.

Keywords: antibiotics, Bacillus subtilis, inhibition, Syzygium cumini

Procedia PDF Downloads 188