Search results for: antibacterial peptides
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 693

Search results for: antibacterial peptides

183 Quantitative Structure-Property Relationship Study of Base Dissociation Constants of Some Benzimidazoles

Authors: Sanja O. Podunavac-Kuzmanović, Lidija R. Jevrić, Strahinja Z. Kovačević

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Benzimidazoles are a group of compounds with significant antibacterial, antifungal and anticancer activity. The studied compounds consist of the main benzimidazole structure with different combinations of substituens. This study is based on the two-dimensional and three-dimensional molecular modeling and calculation of molecular descriptors (physicochemical and lipophilicity descriptors) of structurally diverse benzimidazoles. Molecular modeling was carried out by using ChemBio3D Ultra version 14.0 software. The obtained 3D models were subjected to energy minimization using molecular mechanics force field method (MM2). The cutoff for structure optimization was set at a gradient of 0.1 kcal/Åmol. The obtained set of molecular descriptors was used in principal component analysis (PCA) of possible similarities and dissimilarities among the studied derivatives. After the molecular modeling, the quantitative structure-property relationship (QSPR) analysis was applied in order to get the mathematical models which can be used in prediction of pKb values of structurally similar benzimidazoles. The obtained models are based on statistically valid multiple linear regression (MLR) equations. The calculated cross-validation parameters indicate the high prediction ability of the established QSPR models. This study is financially supported by COST action CM1306 and the project No. 114-451-347/2015-02, financially supported by the Provincial Secretariat for Science and Technological Development of Vojvodina.

Keywords: benzimidazoles, chemometrics, molecular modeling, molecular descriptors, QSPR

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182 Production of Novel Antibiotics of Tylosin by Importing eryK and eryG Genes in Streptomyces fradiae

Authors: Neda Gegar Goshe, M. Moradi, Hossein Rassi

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The antibacterial properties of macrolide antibiotics (such as erythromycin and tylosin) depend ultimately on the glycosylation of otherwise inactive polyketide lactones. Among the sugars commonly found in such macrolides are various 6-deoxyhexoses including the 3-dimethylamino sugars mycaminose and desosamine (4-deoxymycaminose). Some macrolides (such as tylosin) possess multiple sugar moieties, whereas others (such as erythromycin) have two sugar substituents. Streptomyces fradiae is an ideal host for development of generic polyketide-overproducing strains because it contains three of the most common precursors-malonyl-CoA, methylmalonyl-CoA and ethylmalonyl-CoA-used by modular PKS, and is a host that is amenable to genetic manipulation. As patterns of glycosylation markedly influence a macrolide's drug activity, there is considerable interest in the possibility of using combinatorial biosynthesis to generate new pairings of polyketide lactones with sugars, especially 6-deoxyhexoses. Here, we report a successful attempt to alter the aminodeoxyhexose-biosynthetic capacity of Streptomyces fradiae (a producer of tylosin) by importing genes from the erythromycin producer Saccharopolyspora erythraea. The bio transformation of erythromycin-D into the desired major component erythromycin-A involves two final enzymatic reactions, EryK-catalyzed hydroxylation at the C-12 position of the aglycone and EryG-catalyzed O methylation at the C-3 position of macrose. This engineered S. fradiae produced substantial amounts of two potentially useful macrolides that had not previously been obtained by fermentation.

Keywords: tylosin, eryK and eryG genes, streptomyces fradiae

Procedia PDF Downloads 330
181 Liposomal Encapsulation of Silver Nanoparticle for Improved Delivery and Enhanced Anticancer Properties

Authors: Azeez Yusuf, Alan Casey

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Silver nanoparticles (AgNP) are one of the most widely investigated metallic nanoparticles due to their promising antibacterial activities. In recent years, AgNP research has shifted beyond antimicrobial use to potential applications in the medical arena. This shift coupled with the extensive commercial applications of AgNP will further increase human exposure, and the subsequent risk of adverse effects that may result from repeated exposures and inefficient delivery meaning research into improved AgNP delivery is of paramount importance. In this study, AgNP were encapsulated in a natural bio-surfactant, dipalmitoylphosphatyidyl choline (DPPC), in an attempt to enhance the intracellular delivery and simultaneously mediate the associated cytotoxicity of the AgNP. It was noted that as a result of the encapsulation, liposomal-AgNP (Lipo-AgNP) at 0.625 μg/ml induced significant cell death in THP1 cell lines a notably lower dose than that of the uncoated AgNP induced cytotoxicity. The induced cytotoxicity was shown to result in an increased level of DNA fragmentation resulting in a cell cycle interruption at the S phase of the cell cycle. It was shown that the predominate form of cell death upon exposure to both uncoated and Lipo-AgNP was apoptosis, however, a ROS-independent activation of the executioner caspases 3/7 occurred when exposed to the Lipo-AgNP. These findings showed that encapsulation of AgNP enhances AgNP cytotoxicity and mediates an ROS-independent induction of apoptosis.

Keywords: silver nanoparticles, AgNP, cytotoxicity, encapsulation, liposome

Procedia PDF Downloads 127
180 Advanced Textiles for Soldier Clothes Based on Coordination Polymers

Authors: Hossam E. Emam

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The functional textiles development history in the military field could be ascribed as a uniquely interesting research topic. Soldiers are like a high-performance athletes, where monitoring their physical and physiological capabilities is a vital requirement. Functional clothes represent a “second skin” that has a close, “intimate” relationship with the human body. For the application of textiles in military purposes, which is normally required in difficult weather and environmental conditions, several functions are required. The requirements for designing functional military textiles for soldier's protection can be categorized into three categories; i) battle field (protection from chemical warfare agents, flames, and thermal radiation), ii) environmental (water proof, air permeable, UV-protection, antibacterial), iii) physiological (minimize heat stress, low weight, insulative, durability). All of these requirements are important, but the means to fulfill these requirements are not simple and straight forward. Additionally, the combination of more than one function is reported to be very expensive and requires many complicated steps, and the final product is found to be low durability. Not only do all of these requirements are overlapping, but they are also contradicting each other at various levels. Thus, we plan to produce multi-functional textiles (e.g., anti-microbial, UV-protection, fire retardant, photoluminescent) to be applied in military clothes. The current project aims to use quite a simple and applicable technique through the modification of textiles with different coordination polymers and functionalized coordination polymers.

Keywords: functional textiles, military clothes, coordination polymers, antimicrobial, fire retardant, photolumenscent

Procedia PDF Downloads 164
179 Designing Modified Nanocarriers Containing Selenium Nanoparticles Extracted from the Lactobacillus acidophilus and Their Anticancer Properties

Authors: Mahnoosh Aliahmadi, Akbar Esmaeili

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This study synthesized new modified imaging nanocapsules (NCs) of gallium@deferoxamine/folic acid/chitosan/polyaniline/polyvinyl alcohol (Ga@DFA/FA/CS/PANI/PVA) containing Morus nigra extract by selenium nanoparticles prepared from Lactobacillus acidophilus. Se nanoparticles were then deposited on (Ga@DFA/FA/CS/PANI/PVA) using the impregnation method. The modified contrast agents were mixed with M. nigra extract, and their antibacterial activities were investigated by applying them to L929 cell lines. The influence of variable factors including surfactant, solvent, aqueous phase, pH, buffer, minimum Inhibitory concentration (MIC), minimum bactericidal concentration (MBC), cytotoxicity on cancer cells, antibiotic, antibiogram, release and loading, stirring effect, the concentration of nanoparticle, olive oil, and thermotical methods was investigated. The structure and morphology of the synthesized contrast agents were characterized by zeta potential sizer analysis (ZPS), X-Ray diffraction (XRD), Fourier-transform infrared (FT-IR), and energy-dispersive X-ray (EDX), ultraviolet-visible (UV-Vis) spectra, and scanning electron microscope (SEM). The experimental section was conducted and monitored by response surface methods (RSM) and MTT conversion assay. Antibiogram testing of NCs on Pseudomonas aeruginosa bacteria was successful, and the MIC=2 factor was obtained with a less harmful effect.

Keywords: imaging contrast agent, nanoparticles, response surface method, Lactobacillus acidophilus, selenium

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178 Recent Advancements and Future Trends in the Development of Antimicrobial Edible Films for Food Preservation

Authors: Raana Babadi Fathipour

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Food packaging plays a crucial role in protecting food from unwanted external factors. Antibacterial edible films are a promising option for food packaging due to their biodegradability, environmental friendliness, and safety. This paper reviews recent research progress on antimicrobial edible films, focusing on those made from polysaccharides, proteins, and lipids. Polysaccharides and proteins are the primary components of antimicrobial edible films, while lipids primarily serve as plasticizers and carriers for active substances in composite films. For instance, second-generation liposomes have shown great potential as carriers for antimicrobial substances and other bioactive compounds due to their exceptional stability. Furthermore, this paper analyzes recent advancements and future trends in antimicrobial edible films. One promising direction is the integration of antimicrobial edible film materials with delivery systems, such as nanoemulsion and microencapsulation technologies, to ensure stable loading of bioactive substances. Another emerging area of interest is the development of smart and active packaging that allows consumers to assess the freshness of food products without opening the package. pH-sensitive films and smart fluorescent "on-off" sensors for humidity are currently being explored as materials for smart and active packaging to monitor food product freshness, with further exploration anticipated in the future.

Keywords: antimicrobial edible film, biopolymer, antimicrobial agent, encapsulation, antimicrobial assay

Procedia PDF Downloads 38
177 Determination of Cyclic Citrullinated Peptide Antibodies on Quartz Crystal Microbalance Based Nanosensors

Authors: Y. Saylan, F. Yılmaz, A. Denizli

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Rheumatoid arthritis (RA) which is the most common autoimmune disorder of the body's own immune system attacking healthy cells. RA has both articular and systemic effects.Until now romatiod factor (RF) assay is used the most commonly diagnosed RA but it is not specific. Anti-cyclic citrullinated peptide (anti-CCP) antibodies are IgG autoantibodies which recognize citrullinated peptides and offer improved specificity in early diagnosis of RA compared to RF. Anti-CCP antibodies have specificity for the diagnosis of RA from 91 to 98% and the sensitivity rate of 41-68%. Molecularly imprinted polymers (MIP) are materials that are easy to prepare, less expensive, stable have a talent for molecular recognition and also can be manufactured in large quantities with good reproducibility. Molecular recognition-based adsorption techniques have received much attention in several fields because of their high selectivity for target molecules. Quartz crystal microbalance (QCM) is an effective, simple, inexpensive approach mass changes that can be converted into an electrical signal. The applications for specific determination of chemical substances or biomolecules, crystal electrodes, cover by the thin films for bind or adsorption of molecules. In this study, we have focused our attention on combining of molecular imprinting into nanofilms and QCM nanosensor approaches and producing QCM nanosensor for anti-CCP, chosen as a model protein, using anti-CCP imprinted nanofilms. For this aim, anti-CCP imprinted QCM nanosensor was characterized by Fourier transform infrared spectroscopy, atomic force microscopy, contact angle measurements and ellipsometry. The non-imprinted nanosensor was also prepared to evaluate the selectivity of the imprinted nanosensor. Anti-CCP imprinted QCM nanosensor was tested for real-time detection of anti-CCP from aqueous solution. The kinetic and affinity studies were determined by using anti-CCP solutions with different concentrations. The responses related with mass shifts (Δm) and frequency shifts (Δf) were used to evaluate adsorption properties and to calculate binding (Ka) and dissociation (Kd) constants. To show the selectivity of the anti-CCP imprinted QCM nanosensor, competitive adsorption of anti-CCP and IgM was investigated.The results indicate that anti-CCP imprinted QCM nanosensor has a higher adsorption capabilities for anti-CCP than for IgM, due to selective cavities in the polymer structure.

Keywords: anti-CCP, molecular imprinting, nanosensor, rheumatoid arthritis, QCM

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176 Bifidobacterium lactis Fermented Milk Was Not Effective to Eradication of Helicobacter Pylori Infection: A Prospective, Randomized, Double-Blind, Controlled Study

Authors: R. C. Barbuti, M. N. Oliveira, N. P. Perina, C. Haro, P. Bosch, C. S. Bogsan, J. N. Eisig, T. Navarro-Rodriguez

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Background: The management of Helicobacter pylori (H. pylori) eradication is still a matter of discussion, full effectiveness is rarely achieved and it has many adverse effects. Probiotics are believed to have a role in eradicating and possibly preventing H. pylori infection as an adjunctive treatment. The present clinical study was undertaken to see the efficacy of a specially designed fermented milk product containing Bifidobacterium lactis B420 on the eradication of H. pylori infection in a prospective, randomized, double-blind, controlled study in humans. Method: Four test products were specially designed fermented milks, counts of viable cells in all products were 1010 Log CFU. 100 mL-1 for Bifidobacterium lactis-Bifidobacterium species 420, and 1011 Log CFU. 100 mL-1 for Streptococcus thermophiles were administered to subjects infected with H. pylori with a previous diagnosis of functional dyspepsia according to the Rome III criteria in a prospective, randomized, double-blind, placebo-controlled study in humans. Results: After FM supplementation, not all subjects showed a reduction in H. pylori colonization. Conclusion: Bifidobacterium lactis B420, administered twice a day for 90 days did not show an increase in H. pylori eradication effectiveness in Brazilian patients with functional dyspepsia.

Keywords: antibacterial therapy, Bifidobacteria fermented milk, Helicobacter pylori, probiotics

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175 Design of Liquid Crystal Based Interface to Study the Interaction of Gram Negative Bacterial Endotoxin with Milk Protein Lactoferrin

Authors: Dibyendu Das, Santanu Kumar Pal

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Milk protein lactoferrin (Lf) exhibits potent antibacterial activity due to its interaction with Gram-negative bacterial cell membrane component, lipopolysaccharide (LPS). This paper represents fabrication of new Liquid crystals (LCs) based biosensors to explore the interaction between Lf and LPS. LPS self-assembled at aqueous/LCs interface and orients interfacial nematic 4-cyano-4’- pentylbiphenyl (5CB) LCs in a homeotropic fashion (exhibiting dark optical image under polarized optical microscope). Interestingly, on the exposure of Lf on LPS decorated aqueous/LCs interface, an optical image of LCs changed from dark to bright indicating an ordering alteration of interfacial LCs from homeotropic to tilted/planar state. The ordering transition reflects strong binding between Lf and interfacial LPS that, in turn, perturbs the orientation of LCs. With the help of epifluorescence microscopy, we further affirmed the interfacial LPS-Lf binding event by imaging the presence of FITC tagged Lf at the LPS laden aqueous/LCs interface. Finally, we have investigated the conformational behavior of Lf in solution as well as in the presence of LPS using Circular Dichroism (CD) spectroscopy and further reconfirmed with Vibrational Circular Dichroism (VCD) spectroscopy where we found that Lf undergoes alpha-helix to random coil-like structure in the presence of LPS. As a whole the entire results described in this paper establish a robust approach to envisage the interaction between LPS and Lf through the ordering transitions of LCs at aqueous/LCs interface.

Keywords: endotoxin, interface, lactoferrin, lipopolysaccharide

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174 An Investigation of How Salad Rocket May Provide Its Own Defence Against Spoilage Bacteria

Authors: Huda Aldossari

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Members of the Brassicaceae family, such as rocket species, have high concentrations of glucosinolates (GLSs). GSLs and isothiocyanates (ITCs), the product of GLSs hydrolysis, are the most influential compounds that affect flavour in rocket species. Aside from their contribution to the flavour, GSLs and ITCs are of particular interest due to their potential ability to inhibit the growth of human pathogenic bacteria such as E. coli O157. Quantitative and qualitative analysis of glucosinolate compounds in rocket extracts was obtained by Liquid Chromatography-Mass Spectrometry (LC–MS).Each individual component of non-volatile GLSs and ITCs was isolated by High-Performance Liquid Chromatography (HPLC) fractionation. The identity and purity of each fraction were confirmed using Ultra High-Performance Liquid Chromatography (UPLC). The separation of glucosinolates in the complex rocket extractions was performed by optimizing a HPLC fractionation method through changing the mobile phase composition, solvent gradient, and the flow rate. As a result, six glucosinolates compounds (Glucosativin, 4-Methoxyglucobrassicin, Glucotropaeolin GTP, Glucoiberin GIB, Diglucothiobenin, and Sinigrin) have been isolated, identified and quantified in the complex samples. This step aims to evaluate the antibacterial activity of glucosinolates and their enzymatic hydrolysis against bacterial growth of E.coli k12. Therefore, fractions from this study will be used to determine the most active compounds by investigating the efficacy of each component of GLSs and ITCs at inhibiting bacterial growth.

Keywords: rocket, glucosinolates, E.coli k12., HPLC fractionatio

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173 Multifunctional Polydopamine-Silver-Polydopamine Nanofilm With Applications in Digital Microfluidics and SERS

Authors: Yilei Xue, Yat-Hing Ham, Wenting Qiu, Wan Chan, Stefan Nagl

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Polydopamine (PDA) is a popular material in biological and medical applications due to its excellent biocompatibility, outstanding physicochemical properties, and facile fabrication. In this project, a new sandwich-structured PDA and silver (Ag) hybrid material named PDA-Ag-PDA was synthesized and characterized layer-by-layer, where silver nanoparticles (Ag NPs) are wrapped in PDA coatings, using SEM, AFM, 3D surface metrology, and contact angle meter. The silver loading capacity is positively proportional to the roughness value of the initial PDA film. This designed film was subsequently integrated within a digital microfluidic (DMF) platform coupling with an oxygen sensor layer for on-chip antibacterial assay. The concentration of E. coli was quantified on DMF by real-time monitoring oxygen consumption during E. coli growth with the optical oxygen sensor layer. The PDA-Ag-PDA coating shows an 99.9% reduction in E. coli population under non-nutritive condition with 1-hour treatment and has a strong growth inhibition of E. coliin nutrient LB broth as well. Furthermore, PDA-Ag-PDA film maintaining a low cytotoxicity effect to human cells. After treating with PDA-Ag-PDA film for 24 hours, 82% HEK 293 and 86% HeLa cells were viable. The SERS enhancement factor of PDA-Ag-PDA is estimated to be 1.9 × 104 using Rhodamine 6G (R6G). Multifunctional PDA-Ag-PDA coating provides an alternative platform to conjugate biomolecules and perform biological applications on DMF, in particular, for the adhesive protein and cell study.

Keywords: polydopamine, silver nanoparticles, digital microfluidic, optical sensor, antimicrobial assay, SERS

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172 Cocoa Stimulates the Production Bioactive Components of Lactobacillus Casei and Competitively Excludes Foodborne Pathogens

Authors: Mengfei Peng, Serajus Salaheen, Debabrata Biswas

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Lactobacillus casei found in the human intestine and mouth is commonly applied for dairy production. Recently, it was found that some byproducts produced by Lactobacillus exhibited antimicrobial activities against multiple bacteria. Meanwhile, introduction of prebiotic-like foods (e.g. cocoa) or probiotics or both of them as food supplements in human diets as well as in farm animal feeds is believed to be an effective ways in control/reduce the colonization of foodborne bacterial pathogens infection in the gut environment. We hypothesized that cocoa may stimulate the production antimicrobial components of Lactobacillus casei and may potentially inhibit/reduce the colonization and infection of foodborne bacterial pathogens in the gut. Mixed culture of L. casei (LC) with enterohemorrhagic E. coli EDL933 (EHEC), Salmonella Typhimurium LT2 (ST), or Listeria monocytogenes LM2 (LM) showed that LC could competitively exclude (100%) them within 72 h. Further, investigation of cell-free culture supernatant (CFCS) revealed that the antimicrobial effects of LC came from CFCS. CFCS of LC eliminated (100%) EHEC, ST, and LM within 72 h, and 2 h CFCS treatment increased the hydrophobicity of EHEC (5.10 folds), ST (8.48 folds), and LM (2.03 folds). In addition, LC cells exhibited more inhibitive effects than CFCS on cell adhesive and invasive activities of EHEC (52.14% & 90.45%), ST (66.89% & 93.83%), and LM (61.10% & 83.40%). Two clusters of poly-peptides in CFCS were identified by SDS-PAGE, the molecular weights of which are ≈5 KD and 40-45 KD. LC CFCS with overnight growth in the presence of 3% strengthened all of the antimicrobial activities (growth inhibition, outer membrane disruption, and cell infective ability reduction). Liquid chromatography/Mass spectrometry analysis detected 5 unique components in class of flavonoids in LC CFCS with overnight 3% cocoa supplement. Furthermore, qPCR results showed that CFCSs up-regulated the expression level of genes responsible for flagellin synthesis and motility, but down-regulated genes for specific binding and invasion-associated proteins synthesis. The stimulatory effects of cocoa in producing bioactive components of probiotics may aid prevention of foodborne illness caused by major foodborne enteric bacterial pathogens.

Keywords: foodborne pathogens, probiotics, prebiotics, pathogen exclusion

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171 Statistical Pattern Recognition for Biotechnological Process Characterization Based on High Resolution Mass Spectrometry

Authors: S. Fröhlich, M. Herold, M. Allmer

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Early stage quantitative analysis of host cell protein (HCP) variations is challenging yet necessary for comprehensive bioprocess development. High resolution mass spectrometry (HRMS) provides a high-end technology for accurate identification alongside with quantitative information. Hereby we describe a flexible HRMS assay platform to quantify HCPs relevant in microbial expression systems such as E. Coli in both up and downstream development by means of MVDA tools. Cell pellets were lysed and proteins extracted, purified samples not further treated before applying the SMART tryptic digest kit. Peptides separation was optimized using an RP-UHPLC separation platform. HRMS-MSMS analysis was conducted on an Orbitrap Velos Elite applying CID. Quantification was performed label-free taking into account ionization properties and physicochemical peptide similarities. Results were analyzed using SIEVE 2.0 (Thermo Fisher Scientific) and SIMCA (Umetrics AG). The developed HRMS platform was applied to an E. Coli expression set with varying productivity and the corresponding downstream process. Selected HCPs were successfully quantified within the fmol range. Analysing HCP networks based on pattern analysis facilitated low level quantification and enhanced validity. This approach is of high relevance for high-throughput screening experiments during upstream development, e.g. for titer determination, dynamic HCP network analysis or product characterization. Considering the downstream purification process, physicochemical clustering of identified HCPs is of relevance to adjust buffer conditions accordingly. However, the technology provides an innovative approach for label-free MS based quantification relying on statistical pattern analysis and comparison. Absolute quantification based on physicochemical properties and peptide similarity score provides a technological approach without the need of sophisticated sample preparation strategies and is therefore proven to be straightforward, sensitive and highly reproducible in terms of product characterization.

Keywords: process analytical technology, mass spectrometry, process characterization, MVDA, pattern recognition

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170 Surpassing Antibiotic Resistance through Synergistic Effects of Polyethyleneimine-Silver Nanoparticle Complex Coated Mesoporous Silica Trio-Nanoconstructs

Authors: Ranjith Kumar Kankala, Wei-Zhi Lin, Chia-Hung Lee

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Antibiotic resistance in bacteria has become an emergency situation clinically. To improve the efficacy of antibiotics in resistant strains, advancement of nanoparticles is inevitable than ever. Herewith, we demonstrate a design by immobilizing tetracycline (TET) in copper substituted mesoporous silica nanoparticles (Cu-MSNs) through a pH-sensitive coordination link, enabling its release in the acidic environment. Subsequently, MSNs are coated with silver nanoparticles stabilized polyethyleneimine (PEI-SNP) to act against drug-resistant (MDR) bacterial strains. Silver ions released from SNP are capable of sensitizing the resistant strains and facilitate the generation of free radicals capable of damaging the cell components. In addition, copper ions in the framework are also capable of generating free radicals through Fenton-like reaction. Furthermore, the nanoparticles are well-characterized physically, and various antibacterial efficacious tests against isolated multidrug resistant bacterial strain were highly commendable. However, this formulation has no significant toxic effect on normal mammalian fibroblast cells accounting its high biocompatibility. These MSN trio-hybrids, i.e., SNP, tetracycline, and copper ions result in synergistic effects, and their advancement could bypass resistance and allow synergism for effective treatment of antibiotic clinically.

Keywords: antibiotic resistance, copper, mesoporous silica nanoparticles, Ph-sensitive release, polyethyleneimine, silver, tetracycline

Procedia PDF Downloads 178
169 Targeting APP IRE mRNA to Combat Amyloid -β Protein Expression in Alzheimer’s Disease

Authors: Mateen A Khan, Taj Mohammad, Md. Imtaiyaz Hassan

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Alzheimer’s disease is characterized by the accumulation of the processing products of the amyloid beta peptide cleaved by amyloid precursor protein (APP). Iron increases the synthesis of amyloid beta peptides, which is why iron is present in Alzheimer's disease patients' amyloid plaques. Iron misregulation in the brain is linked to the overexpression of APP protein, which is directly related to amyloid-β aggregation in Alzheimer’s disease. The APP 5'-UTR region encodes a functional iron-responsive element (IRE) stem-loop that represents a potential target for modulating amyloid production. Targeted regulation of APP gene expression through the modulation of 5’-UTR sequence function represents a novel approach for the potential treatment of AD because altering APP translation can be used to improve both the protective brain iron balance and provide anti-amyloid efficacy. The molecular docking analysis of APP IRE RNA with eukaryotic translation initiation factors yields several models exhibiting substantial binding affinity. The finding revealed that the interaction involved a set of functionally active residues within the binding sites of eIF4F. Notably, APP IRE RNA and eIF4F interaction were stabilized by multiple hydrogen bonds with residues of APP IRE RNA and eIF4F. It was evident that APP IRE RNA exhibited a structural complementarity that tightly fit within binding pockets of eIF4F. The simulation studies further revealed the stability of the complexes formed between RNA and eIF4F, which is crucial for assessing the strength of these interactions and subsequent roles in the pathophysiology of Alzheimer’s disease. In addition, MD simulations would capture conformational changes in the IRE RNA and protein molecules during their interactions, illustrating the mechanism of interaction, conformational change, and unbinding events and how it may affect aggregation propensity and subsequent therapeutic implications. Our binding studies correlated well with the translation efficiency of APP mRNA. Overall, the outcome of this study suggests that the genomic modification and/or inhibiting the expression of amyloid protein by targeting APP IRE RNA can be a viable strategy to identify potential therapeutic targets for AD and subsequently be exploited for developing novel therapeutic approaches.

Keywords: Alzheimer's disease, Protein-RNA interaction analysis, molecular docking simulations, conformational dynamics, binding stability, binding kinetics, protein synthesis.

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168 Synthesis and Preparation of Carbon Ferromagnetic Nanocontainers for Cancer Therapy

Authors: L. Szymanski, Z. Kolacinski, Z. Kamiński, G. Raniszewski, J. Fraczyk, L. Pietrzak

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In the article the development and demonstration of method and the model device for hyperthermic selective destruction of cancer cells are presented. This method was based on the synthesis and functionalization of carbon nanotubes serving as ferromagnetic material nano containers. Methodology of the production carbon - ferromagnetic nanocontainers includes: the synthesis of carbon nanotubes, chemical and physical characterization, increasing the content of ferromagnetic material and biochemical functionalization involving the attachment of the key addresses. Biochemical functionalization of ferromagnetic nanocontainers is necessary in order to increase the binding selectively with receptors presented on the surface of tumour cells. Multi-step modification procedure was finally used to attach folic acid on the surface of ferromagnetic nanocontainers. Folic acid is ligand of folate receptors which is overexpresion in tumor cells. The presence of ligand should ensure the specificity of the interaction between ferromagnetic nanocontainers and tumor cells. The chemical functionalization contains several step: oxidation reaction, transformation of carboxyl groups into more reactive ester or amide groups, incorporation of spacer molecule (linker), attaching folic acid. Activation of carboxylic groups was prepared with triazine coupling reagent (preparation of superactive ester attached on the nanocontainers). The spacer molecules were designed and synthesized. In order to ensure biocompatibillity of linkers they were built from amino acids or peptides. Spacer molecules were synthesized using the SPPS method. Synthesis was performed on 2-Chlorotrityl resin. The linker important feature is its length. Due to that fact synthesis of peptide linkers containing from 2 to 4 -Ala- residues was carried out. Independent synthesis of the conjugate of foilic acid with 6-aminocaproic acid was made. Final step of synthesis was connecting conjugat with spacer molecules and attaching it on the ferromagnetic nanocontainer surface. This article contains also information about special CVD and microvave plasma system to produce nanotubes and ferromagnetic nanocontainers. The first tests in the device for hyperthermal RF generator will be presented. The frequency of RF generator was in the ranges from 10 to 14Mhz and from 265 to 621kHz.

Keywords: synthesis of carbon nanotubes, hyperthermia, ligands, carbon nanotubes

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167 Determination of the Effect of Kaolin on the Antimicrobial Activity of Metronidazole-Kaolin Interaction

Authors: Omaimah Algohary

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Kaolin is one of the principle intestinal adsorbents, has traditionally been used internally in the treatment of various enteric disorders, colitis, enteritis, dysentery, and diarrhea associated with food and alkaloidal poisoning and in traveler’s diarrhea. It binds to and traps bacteria and its toxins and gases in the gut. It also binds to water in the gut, which helps to make the stools firmer, hence giving symptomatic relief. Metronidazole is a synthetic antibacterial agent that is used primarily in the treatment of various anaerobic infections such as intra-abdominal infections, antiprotozoal, and as amebicidal. The need for safe, therapeutically effective antidiarrheal combination continuously lead to effective treatment. Metronidazol used for treatment of anaerobic bacteria and kaolin , when administered simultaneously, Metronidazole–Kaolin interactions have been reported by FDA but not studied. This project is the first to study the effect of Metronidazole–Kaolin interactions on the antimicrobial activity of metronidazole. Agar diffusion method performed to test the antimicrobial activity of metronidazole–kaolin antidiarrheal combination from aqueous solutions at an in-vivo simulated pHs conditions that obtained at 37+0.5 °C on Helicobacter pylori as anaerobic bacteria and E.coli as aerobic bacteria and used as a control for the technique. The antimicrobial activity of metronidazole combination as 1:1 and 1:2 with kaolin was abolished in acidic media as no zones of inhibition shown compared to only metronidazole that used as a control. In alkaline media metronidazole combination as 1:1 and 1:2 with kaolin showed diminutive activity compared to the control. These results proved that the kaolin adsorb metronidazole and abolish its antimicrobial activity and such combination should be avoided.

Keywords: kaolin, metronidazole, interaction, Helicobacter pylori. E. coli, antimicrobial activity

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166 Preservation of Traditional Algerian Sausage Against Microbial Activity by the Garlic (Allium Sativum L.)

Authors: Abed Hannane, Rouag Noureddine

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The present study aims to evaluate the association of fresh garlic (Allium sativum L.) and storage at 4°C in preserving the microbiological, nutritional, and sanitary quality of Merguez-type sausages prepared and sold locally from meat offal. The analysis focused on the evaluation of the microbiological quality of fifteen samples randomly taken from several butcheries in the wilaya of BBA, eastern Algeria. The bacteriological analysis revealed the presence of 6.88.10⁵ CFU/g of total aerobic bacteria, 5.39.10⁵ CFU/g of total coliforms, 2.23.10⁵ CFU/g of fecal coliforms, 2.43.103 CFU/g of Escherichia coli and 1.8.10⁵ CFU/g of coagulase-positive staphylococci, values higher than Algerian standards. The addition of fresh garlic as an antibacterial preservative at concentrations of 0.06, 0.12, 0.18, and 0.24 g/g to ground beef samples and stored in the refrigerator at 4°C for 15 days. The addition of garlic to Merguez made it possible to significantly reduce the presence of different bacterial groups during their refrigerated storage, compared to untreated meat, bringing it below the standards defined in the matter. Thus, the use of garlic as a food additive at a concentration of 0.12 g/g was sufficient to obtain levels according to Algerian standards equal to 1.8.10⁴ CFU/g of total aerobic bacteria, 9.48.10³ CFU/ g of total coliforms, 3.68.10³ UFC/g fecal coliforms, 4.56.10² UFC/g of E.coli 2.39.10⁴ UFC/g of coagulase-positive staphylococci. It is clear that thanks to the addition of garlic to Merguez, the sanitary quality has been improved by reducing the aerobic bacterial load and increasing the shelf life at 4°C.

Keywords: antimicrobial effect, garlic, sausage, storage

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165 Assessment of the Role of Plasmid in Multidrug Resistance in Extended Spectrum βEtalactamase Producing Escherichia Coli Stool Isolates from Diarrhoeal Patients in Kano Metropolis Nigeria

Authors: Abdullahi Musa, Yakubu Kukure Enebe Ibrahim, Adeshina Gujumbola

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The emergence of multidrug resistance in clinical Escherichia coli has been associated with plasmid-mediated genes. DNA transfer among bacteria is critical to the dissemination of resistance. Plasmids have proved to be the ideal vehicles for dissemination of resistance genes. Plasmids coding for antibiotic resistance were long being recognized by many researchers globally. The study aimed at determining the antibiotic susceptibility pattern of ESBL E. coli isolates claimed to be multidrug resistance using disc diffusion method. Antibacterial activity of the test isolates was carried out using disk diffusion methods. The results showed that, majority of the multidrug resistance among clinical isolates of ESBL E. coli was as a result of acquisition of plasmid carrying antibiotic-resistance genes. Production of these ESBL enzymes by these organisms which are normally carried by plasmid and transfer from one bacterium to another has greatly contributed to the rapid spread of antibiotic resistance amongst E. coli isolates, which lead to high economic burden, increase morbidity and mortality rate, complication in therapy and limit treatment options. To curtail these problems, it is of significance to checkmate the rate at which over the counter drugs are sold and antibiotic misused in animal feeds. This will play a very important role in minimizing the spread of resistance bacterial strains in our environment.

Keywords: Escherichia coli, plasmid, multidrug resistance, ESBL, pan drug resistance

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164 Engineering C₃ Plants with SbtA, a Cyanobacterial Transporter, for Enhancing CO₂ Fixation

Authors: Vandana Deopanée Tomar, Gurpreet Kaur Sidhu, Panchsheela Nogia, Rajesh Mehrotra, Sandhya Mehrotra

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The cyanobacterial CO₂ concentrating mechanism (CCM) operates to raise the levels of CO₂ in the vicinity of the main carboxylation enzyme Rubisco which is encapsulated in protein micro compartments called carboxysomes. Thus, due to the presence of CCM, cyanobacterial cells are able to work with high photosynthetic efficiency even at low Ci conditions and can accumulate 1000 folds high internal concentrations of Ci than external environment. Engineering of some useful CCM components into higher plants is one of the plausible approaches to improve their photosynthetic performance. The first step and the simplest approach for attaining this objective would be the transfer of cyanobacterial bicarbonate transporter such as SbtA to inner chloroplast envelope of C₃ plants. For this, SbtA transporter gene from Synechococcus elongatus PCC 7942 was fused to a transit peptide element to generate chimeric constructs in order to direct it to chloroplast inner envelope. Two transit peptides namely, TnaXTP (transit peptide from AT3G56160) and TMDTP (transit peptide from AT2G02590) were shortlisted from Arabidopsis thaliana genome and cloned in plant expression vector pCAMBIA1302 having mgfp5 as a reporter gene. Plant transformation was done by agro infiltration and Agrobacterium mediated co-culture. DNA, RNA, and protein were isolated from the leaves four days post infiltration, and the presence of transgene was confirmed by gene specific PCR (Polymerase Chain Reaction) analysis and by RT-PCR (Reverse Transcription Polymerase Chain Reaction). The expression was confirmed at the protein level by western blotting using anti-GFP primary antibody and horseradish peroxidase (HRP) conjugated secondary antibody. The localization of the protein was detected by confocal microscopy of isolated protoplasts. We observed chloroplastic expression for both the fusion constructs which suggest that the transit peptide sequences are capable of taking the cargo protein to the chloroplasts. These constructs are now being used to generate stable transgenic plants by Agrobacterium mediated transformation. The stability of transgene expression will be analyzed from T₀ to T₂ generation.

Keywords: agro infiltration, bicarbonate transporter, carbon concentrating mechanisms, cyanobacteria, SbtA

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163 Antibiogram Profile of Antibacterial Multidrug Resistance in Democratic Republic of Congo: Situation in Bukavu City Hospitals

Authors: Justin Ntokamunda Kadima, Christian Ahadi Irenge, Patient Birindwa Mulashe, Félicien Mushagalusa Kasali, Patient Wimba

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Background: Bacterial strains carrying multidrug resistance traits are gaining ground worldwide, especially in countries with limited resources. This study aimed to evaluate the spreading of multidrug-resistant bacteria strains in Bukavu city hospitals in the Democratic Republic of Congo. Methods: We analyzed 758 antibiogram data recorded in files of patients consulted between January 2016 and December 2017 at three reference hospitals selected as sentinel sites, namely the Panzi General Reference Hospital (HGP), BIO -PHARM hospital (HBP), and Saint Luc Clinic (CSL). Results: Of 758 isolates tested, the laboratories identified 12 bacterial strains in 712 isolates, of which 223 (29.42%) presented MDR profile, including Escherichia coli (11.48%), Klebsiella pneumonia (6.07%), Enterobacter (5.8%), Staphylococcus aureus and coagulase-negative Staphylococci (1.58%), Proteus mirabilis (1.85%), Salmonella enterica (1.19%), Pseudomonas aeruginosa (0.53%), Streptococcus pneumonia (0.4%)), Citrobacter (0.13%), Neisseria gonorrhea (0.13%), Enterococcus faecalis (0.13%), and Morganella morganii (0.13%). Infected patients were significantly more adults (73.1% vs. 21.5%) compared to children and mainly women (63.7% vs. 30.9%; p = 0.001). Conclusion: The observed expansion requires that hospital therapeutic committees set up an effective clinical management system and define the right combinations of antibiotics.

Keywords: multidrug resistance, bacteria, antibiogram, Bukavu

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162 Bioprospecting of Marine Actinobacteria: The Leading Way for Industrially Important Enzymes and Bioactive Natural Products

Authors: Ramesh Subramani, Mathivanan Narayanasamy, William Aalbersberg

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It is well accepted by last 35 years of research and on-going programmes that marine environment harbours abundant and unique biodiversity, which is currently playing as an important source in bioprospecting. It has become apparent that marine microorganisms are lead in the biodiscovery. Among marine organisms, actinobacteria are a target phylum for discovering novel antibiotics against increasing the multi-drug resistant human pathogens because of these taxa representing for novel genera and species. Marine actinomycetes are a proven source of new antibiotic leads and novel enzymes with important industrial applications. A total of 183 streptomycete and 25 non-streptomycete strains were isolated from different marine samples collected from north-eastern part of the Indian Ocean. Among them, 111 isolates displayed antibacterial activity against human pathogens and 151 exhibited antifungal activity against phytopathogens. Importantly, most of them produced various extracellular enzymes and 58 of them produced exopolysaccharides. Totally eight small bioactive compounds and a thermostable alkaline protease have been purified from a selected strain, Streptomyces fungicidicus. Besides, our on-going studies on non-streptomycete strains (rare actinomycetes) are most likely promising resource for new and unique compounds against current emerging drug-resistant pathogens. We have just recognised the chemical diversity in marine microorganisms. Therefore it is worthwhile to continue the exploration of marine microorganisms for new drug leads, novel enzymes and other bioprospecting research.

Keywords: bioactive compounds, industrial enzymes, marine actinobacteria, microbial metabolites, marine natural products

Procedia PDF Downloads 255
161 Optimization of the Enzymatic Synthesis of the Silver Core-Shell Nanoparticles

Authors: Lela Pintarić, Iva Rezić, Ana Vrsalović Presečki

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Considering an enormous increase of the use of metal nanoparticles with the exactly defined characteristics, the main goal of this research was to found the optimal and environmental friendly method of their synthesis. The synthesis of the inorganic core-shell nanoparticles was optimized as a model. The core-shell nanoparticles are composed of the enzyme core belted with the metal ions, oxides or salts as a shell. In this research, enzyme urease was the core catalyst and the shell nanoparticle was made of silver. Silver nanoparticles are widespread utilized and some of their common uses are: as an addition to disinfectants to ensure an aseptic environment for the patients, as a surface coating for neurosurgical shunts and venous catheters, as an addition to implants, in production of socks for diabetics and athletic clothing where they improve antibacterial characteristics, etc. Characteristics of synthesized nanoparticles directly depend on of their size, so the special care during this optimization was given to the determination of the size of the synthesized nanoparticles. For the purpose of the above mentioned optimization, sixteen experiments were generated by the Design of Experiments (DoE) method and conducted under various temperatures, with different initial concentration of the silver nitrate and constant concentration of the urease of two separate manufacturers. Synthesized nanoparticles were analyzed by the Nanoparticle Tracking Analysis (NTA) method on Malvern NanoSight NS300. Results showed that the initial concentration of the silver ions does not affect the concentration of the synthesized silver nanoparticles neither their size distribution. On the other hand, temperature of the experiments has affected both of the mentioned values.

Keywords: core-shell nanoparticles, optimization, silver, urease

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160 Sensing Endocrine Disrupting Chemicals by Virus-Based Structural Colour Nanostructure

Authors: Lee Yujin, Han Jiye, Oh Jin-Woo

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The adverse effects of endocrine disrupting chemicals (EDCs) has attracted considerable public interests. The benzene-like EDCs structure mimics the mechanisms of hormones naturally occurring in vivo, and alters physiological function of the endocrine system. Although, some of the most representative EDCs such as polychlorinated biphenyls (PCBs) and phthalates compounds already have been prohibited to produce and use in many countries, however, PCBs and phthalates in plastic products as flame retardant and plasticizer are still circulated nowadays. EDCs can be released from products while using and discarding, and it causes serious environmental and health issues. Here, we developed virus-based structurally coloured nanostructure that can detect minute EDCs concentration sensitively and selectively. These structurally coloured nanostructure exhibits characteristic angel-independent colors due to the regular virus bundle structure formation through simple pulling technique. The designed number of different colour bands can be formed through controlling concentration of virus solution and pulling speed. The virus, M-13 bacteriophage, was genetically engineered to react with specific ECDs, typically PCBs and phthalates. M-13 bacteriophage surface (pVIII major coat protein) was decorated with benzene derivative binding peptides (WHW) through phage library method. In the initial assessment, virus-based color sensor was exposed to several organic chemicals including benzene, toluene, phenol, chlorobenzene, and phthalic anhydride. Along with the selectivity evaluation of virus-based colour sensor, it also been tested for sensitivity. 10 to 300 ppm of phthalic anhydride and chlorobenzene were detected by colour sensor, and showed the significant sensitivity with about 90 of dissociation constant. Noteworthy, all measurements were analyzed through principal component analysis (PCA) and linear discrimination analysis (LDA), and exhibited clear discrimination ability upon exposure to 2 categories of EDCs (PCBs and phthalates). Because of its easy fabrication, high sensitivity, and the superior selectivity, M-13 bacteriophage-based color sensor could be a simple and reliable portable sensing system for environmental monitoring, healthcare, social security, and so on.

Keywords: M-13 bacteriophage, colour sensor, genetic engineering, EDCs

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159 Synthesis of Plant-Mediated Silver Nanoparticles Using Erythrina indica Extract and Evaluation of Their Anti-Microbial Activities

Authors: Chandra Sekhar Singh, P. Chakrapani, B. Arun Jyothi, A. Roja Rani

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The green synthesis of metallic nanoparticles (NPs) involves biocompatible ingredients under physiological conditions of temperature and pressure. Moreover, the biologically active molecules involved in the green synthesis of NPs act as functionalizing ligands, making these NPs more suitable for biomedical applications. Among the most important bioreductants are plant extracts, which are relatively easy to handle, readily available, low cost, and have been well explored for the green synthesis of other nanomaterials. Various types of metallic NPs have already been synthesized using plant extracts. They have wide applicability in various areas such as electronics, catalysis, chemistry, energy, and medicine. Metallic nanoparticles are traditionally synthesized by wet chemical techniques, where the chemicals used are quite often toxic and flammable. In our study, we were described a cost effective and environment friendly technique for green synthesis of silver nanoparticles from 1mM AgNO3 solution through the aqueous extract of Erythrina indica as reducing as well as capping agent. Nanoparticles were characterized using UV–Vis absorption spectroscopy, FTIR, XRD, X-ray diffraction, SEM and TEM analysis showed the average particle size of 30 nm as well as revealed their spherical structure. Further these biologically synthesized nanoparticles were found to be highly toxic against different human pathogens viz. two Gram positive namely Klebsiella pneumonia and Bacillus subtilis bacteria and two were Gram negative bacteria namely Staphylococcus aureus and Escherichia coli (E. coli). This is for the first time reporting that Erythrina indica plant extract was used for the synthesis of nanoparticles.

Keywords: silver nanoparticles, green synthesis, antibacterial activity, FTIR, TEM, SEM

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158 Effects of Caprine Arthritis-Encephalitis Virus (CAEV) Infection on the Expression of Cathelicidin Genes in Goat Blood Leukocytes

Authors: Daria Reczynska, Justyna Jarczak, Michal Czopowicz, Danuta Sloniewska, Karina Horbanczuk, Wieslaw Jarmuz, Jaroslaw Kaba, Emilia Bagnicka

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Since people, animals and plants are constantly exposed to pathogens they have developed very complex systems of defense. Among ca. 1000 antimicrobial peptides from different families so far identified, approximately 30 belonging to cathelicidin family can be found in mammals. Cathelicidins probably constitute the first line of defense because they can act at a physiological salt concentration which is present in healthy tissues. Moreover, the low salt concentration which is present in infected tissues inhibits their activity. In goat bactenecin 7.5 (BAC7.5), bactenecin 5 (BAC5), myeloid antimicrobial peptide 28 (MAP28), myeloid antimicrobial peptide 34 (MAP34 A and B), goat bactenecin3.4 (ChBac3.4) were identified. Caprine arthritis-encephalitis (CAE) caused by small ruminant lentivirus (SRLV) is economic problem. The main CAE symptoms are weight loss, arthritis, pneumonia and mastitis (significant elevation of the somatic cell count and deterioration of some technological parameters). The study was conducted on 24 dairy goats. The animals were divided into two groups: experimental (SRLV-infected) and control (non-infected). The blood samples were collected five times: on the 1st, 7th, 30th, 90th and 150thday of lactation. The levels of transcripts of BAC7.5, BAC5, MAP28 and MAP34 genes in blood leucocytes were measured using qPCR method. There were no differences in mRNA levels of studied genes between stages of lactation. The differences were observed in expressions of BAC5, MAP28 and MAP34 genes with lower levels in the experimental group. There was no difference in BAC7.5 expression between groups. The decreased levels of transcripts of cathelicidin genes in blood leucocytes of SRLV-infected goats may indicate the disturbances of homeostasis in organisms. It can be concluded that SRLV infection seems to inhibit expression of cathelicidin genes. The study was financed by a grant from the National Scientific Center No. UMO-2013/09/B/NZ/03514.

Keywords: goat, CAEV, cathelicidins, blood leukocytes, gene expression

Procedia PDF Downloads 257
157 Effects of Beeswax Coating on the Properties of Cocoa Bean Shell Based Papers

Authors: Sri Rejeki, Tamrin Tamrin, RH. F. Faradilla, Muhammad N. Ibrahim, Mariana M., Irnawati Irnawati

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Cocoa bean shells, despite their antioxidant and antimicrobial properties, are still considered as an underutilized agricultural waste. The functional properties and their lignocelluloses content make cocoa bean shells a potential material for paper-based food packaging. In our previous research, we have successfully produced papers from cocoa bean shells that had antioxidant and antibacterial activities. However, the hydrophilic nature of the lignocelluloses of cocoa bean shells hinders the application of the paper to be used as a food packaging. In this research, we aimed to study the effects of beeswax coating on the wettability and mechanical properties of the paper. The coating was done by dipping the papers in beeswax solution several times and in three different beeswax concentrations. The number of dipping and beeswax concentration significantly (p<0.05) affected the water contact angle of the papers. Results show that the water contact angle increases dramatically due to the coating treatment. The control paper or uncoated paper had a contact angle of 40.50o, while the contact angle of the best-coated paper (D3B3: 3x dipping, 3g/10mL beeswax) reached 96.93o. Both tensile strength and percent elongation were not significantly (p>0.05) affected by the coating treatment. This showed that beeswax was a potential organic material to improve the hydrophobicity of paper from cocoa bean shells without any undesirable effects on the mechanical properties of the paper.

Keywords: cocoa bean shell, paper, beeswax, coating, contact angle

Procedia PDF Downloads 127
156 Phage Capsid for Efficient Delivery of Cytotoxic Drugs

Authors: Simona Dostalova, Dita Munzova, Ana Maria Jimenez Jimenez, Marketa Vaculovicova, Vojtech Adam, Rene Kizek

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The boom of nanomedicine in recent years has led to the development of numerous new nanomaterials that can be used as nanocarriers in the drug delivery. These nanocarriers can either be synthetic or natural-based. The disadvantage of many synthetic nanocarriers is their toxicity in patient’s body. Protein cages that can naturally be found in human body do not exhibit such disadvantage. However, the release of cargo from some protein cages in target cells can be problematic. As a special type of protein cages can serve the capsid of many viruses, including phage. Phages infect bacterial cells; therefore they are not harmful to human cells. The targeting of phage particles to cancer cells can be solved by producing of empty phage capsids during which the targeting moieties (e.g. peptides) can be cloned into genes of phage capsid to decorate its surface. Moreover, the produced capsids do not contain viral nucleic acid and are therefore not infectious to beneficial bacteria in the patient’s body. The protein cage composed of viral capsid is larger than other frequently used apoferritin cage but its size is still small enough to benefit from passive targeting by Enhanced Permeability and Retention effect. In this work, bacteriophage λ was used, both whole and its empty capsid for delivery of different cytotoxic drugs (cisplatin, carboplatin, oxaliplatin, etoposide and doxorubicin). Large quantities of phage λ were obtained from phage λ-producing strain of E. coli cultivated in medium with 0.2 % maltose. After killing of E. coli with chloroform and its removal by centrifugation, the phage was concentrated by ultracentrifugation at 130 000 g and 4 °C for 3 h. The encapsulation of the drugs was performed by infusion method and four different concentrations of the drugs were encapsulated (200; 100; 50; 25 µg/ml). Free molecules of drugs were removed by dialysis. The encapsulation was verified using spectrophotometric and electrochemical methods. The amount of encapsulated drug linearly increased with the amount of applied drug (determination coefficient R2=0.8013). 76% of applied drug was encapsulated in phage λ particles (concentration of 10 µg/ml), even with the highest applied concentration of drugs, 200 µg/ml. Only 1% of encapsulated drug was detected in phage DNA. Similar results were obtained with encapsulation in phage empty capsid. Therefore, it can be concluded that the encapsulation of drugs into phage particles is efficient and mostly occurs by interaction of drugs with protein capsid.

Keywords: cytostatics, drug delivery, nanocarriers, phage capsid

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155 Human Xanthine Oxidase Inhibitory Effect, in vivo Antioxidant Activity of Globularia alypum L. Extracts

Authors: N. Boussoualim, H. Trabsa, I. Krache, S. Aouachria, S. Boumerfeg, L. Arrar, A. Baghiani

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The aim of this study consisted in evaluating the antioxidant in vivo properties, anti-hemolytic and XOR inhibitory effect of Globularia alypum L. (GA) extracts. GA was submitted to extraction and fractionation to give crude (CrE), chloroformique (ChE), ethyle acetate (EAE) and aqueos (AqE) extracts. Total polyphenols contents of GA extracts were determined; EAE is the most rich in polyphenols (157,74±5,27 mg GAE/mg of extract). GA Extracts inhibited XO in a concentration-dependent manner, the EAE showed the highest inhibitory properties on the XOR activity (IC50=0,083±0,001 mg/ml), followed by CrE and ChE. The antioxidant activities of the CrE, EAE, and AqE were tested by an in vivo assay in mice, the plasma ability to inhibit DPPH radical was measured, The CrE was found to exhibit the greatest scavenger activity with 48.41±2.763%, followed by AqE and EAE (40.54±7.51% and 41.79±1.654%, respectively). Total antioxidant capacity of red blood cells was measured, from the kinetics of hemolysis obtained. The calculated HT50 reveal an extension of time for half hemolysis in all treated groups compared with the control group. CrE increase significantly HT50 (112,8±2,427). The hemolysis is lagged, indicating that endogenous antioxidants in the erythrocytes can trap radicals to protect them against free-radical-induced hemolysis. Antimicrobial activities of the extracts were determined by the disc diffusion method. Test microorganisms were; 4 Gram positive, 7 gram negative bacteria, most active extracts were EAE and CrE. We deduce a great relationship between the effect on the extracts antibacterial effect and their contents in flavonoid.

Keywords: Globularia alypum, Xanthine oxidoreductase, in vivo-antioxidant activity, hemolysis, polyphenol

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154 Valorization of Seafood and Poultry By-Products as Gelatin Source and Quality Assessment

Authors: Elif Tugce Aksun Tumerkan, Umran Cansu, Gokhan Boran, Fatih Ozogul

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Gelatin is a mixture of peptides obtained from collagen by partial thermal hydrolysis. It is an important and useful biopolymer that is used in the food, pharmacy, and photography products. Generally, gelatins are sourced from pig skin and bones, beef bone and hide, but within the last decade, using alternative gelatin resources has attracted some interest. In this study, functional properties of gelatin extracted from seafood and poultry by-products were evaluated. For this purpose, skins of skipjack tuna (Katsuwonus pelamis) and frog (Rana esculata) were used as seafood by-products and chicken skin as poultry by-product as raw material for gelatin extraction. Following the extraction of gelatin, all samples were lyophilized and stored in plastic bags at room temperature. For comparing gelatins obtained; chemical composition, common quality parameters including bloom value, gel strength, and viscosity in addition to some others like melting and gelling temperatures, hydroxyproline content, and colorimetric parameters were determined. The results showed that the highest protein content obtained in frog gelatin with 90.1% and the highest hydroxyproline content was in chicken gelatin with 7.6% value. Frog gelatin showed a significantly higher (P < 0.05) melting point (42.7°C) compared to that of fish (29.7°C) and chicken (29.7°C) gelatins. The bloom value of gelatin from frog skin was found higher (363 g) than chicken and fish gelatins (352 and 336 g, respectively) (P < 0.05). While fish gelatin had higher lightness (L*) value (92.64) compared to chicken and frog gelatins, redness/greenness (a*) value was significantly higher in frog skin gelatin. Based on the results obtained, it can be concluded that skins of different animals with high commercial value may be utilized as alternative sources to produce gelatin with high yield and desirable functional properties. Functional and quality analysis of gelatin from frog, chicken, and tuna skin showed by-product of poultry and seafood can be used as an alternative gelatine source to mammalian gelatine. The functional properties, including bloom strength, melting points, and viscosity of gelatin from frog skin were more admirable than that of the chicken and tuna skin. Among gelatin groups, significant characteristic differences such as gel strength and physicochemical properties were observed based on not only raw material but also the extraction method.

Keywords: chicken skin, fish skin, food industry, frog skin, gel strength

Procedia PDF Downloads 139