Search results for: acid lactid bacteria
431 The Importance of Oral Mucosal Biopsy Selection Site in Areas of Field Change: A Case Report
Authors: Timmis W., Simms M., Thomas C.
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This case discusses the management of two floors of mouth (FOM) Squamous Cell Carcinomas (SCC) not identified upon initial biopsy. A 51 year-old male presented with right FOM erythroleukoplakia. Relevant medical history included alcoholic dependence syndrome and alcoholic liver disease. Relevant drug therapy encompassed acamprosate, folic acid, hydroxocobalamin and thiamine. The patient had a 55.5 pack-year smoking history and alcohol dependence from age 14, drinking 16 units/day. FOM incisional biopsy and histopathological analysis diagnosed Carcinoma in situ. Treatment involved wide local excision. Specimen analysis revealed two separate foci of pT1 moderately differentiated SCCs. Carcinoma staging scans revealed no pathological lymphadenopathy, no local invasion or metastasis. SCCs had been excised in completion with narrow margins. MDT discussion concluded that in view of the field changes it would be difficult to identify specific areas needing further excision, although techniques such as Lugol’s Iodine were considered. Further surgical resection, surgical neck management and sentinel lymph node biopsy was offered. The patient declined intervention, primary management involved close monitoring alongside alcohol and smoking cessation referral. Narrow excisional margins can increase carcinoma recurrence risk. Biopsy failed to identify SCCs, despite sampling an area of clinical concern. For gross field change multiple incisional biopsies should be considered to increase chance of accurate diagnosis and appropriate treatment. Coupling of tobacco and alcohol has a synergistic effect, exponentially increasing the relative risk of oral carcinoma development. Tobacco and alcoholic control is fundamental in reducing treatment‑related side effects, recurrence risk and second primary cancer development.Keywords: alcohol dependence, biopsy, oral carcinoma, tobacco
Procedia PDF Downloads 112430 Citrullinated Myelin Basic Protein Mediated Inflammation in Astrocytes
Authors: Lali Shanshiashvili, Marika Chikviladze, Nino Mamulashvili, Maia Sepashvili, Nana Narmania, David Mikeladze
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Purpose: During demyelinating inflammatory diseases and after the damage of the myelin sheet, myelin-derived proteins, including myelin basic protein (MBP), are secreted into the extracellular space. MBP shows extensive post-translational modifications, including the deimination of arginine residues. Deiminated MBP is structurally less ordered, susceptible to proteolytic attack, and more immunogenic than the unmodified one. It is hypothesized that MBP could change the inflammatory response in astrocytes. Methods: MBP was isolated and purified from bovine brain white matter. Primary astrocyte cultures were prepared from whole brains of 2-day-old Wistar rats. For evaluation of glutamate uptake/release in astrocytes following treatment of cells with MBP charge isomers, Glutamate Assay Kit was used. The expression of EAAT-2 (excitatory amino acid transporters), peroxisome proliferator-activated receptor gamma (PPAR- γ), inhibitor of nuclear factor kappa B (IkB), and high mobility group protein B1 (HMGB1) in astrocytes were assayed by Western Blot analysis. Results: This study investigated the action of deiminated isomer (C8) on the cultured primary astrocytes and compared its effects with the effects of unmodified C1 isomers. The study found that C8 and C1 MBP differently act on the uptake and release of glutamate in astrocytes: nonmodified C1 MBP increases the uptake of glutamate and does not change the release, whereas C8 decreases the release of glutamate but does not alter the uptake. Nevertheless, both isomers increased the expression of PPAR-γ and EAAT2 in the same intensity. However, immunostaining and Western Blots of cell lysates showed a decrease of IkB and increased expression of HMGB1 after the treatment of astrocytes by C8. Moreover, in the presence of C8, astrocytes release more nitric oxide than unmodified C1 isomers. Conclusion: These data suggest that the deiminated isomer of MBP evokes an inflammatory response and enhances the ability of astrocytes to release proinflammatory mediators through activation of NF-kB after the breakdown of myelin sheets. Acknowledgment: This research was supported by the SRNSF Georgia RF17_534 grant.Keywords: myelin basic protein, glutamate, deimination, astrocytes, inflammation
Procedia PDF Downloads 205429 Reactive Transport Modeling in Carbonate Rocks: A Single Pore Model
Authors: Priyanka Agrawal, Janou Koskamp, Amir Raoof, Mariette Wolthers
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Calcite is the main mineral found in carbonate rocks, which form significant hydrocarbon reservoirs and subsurface repositories for CO2 sequestration. The injected CO2 mixes with the reservoir fluid and disturbs the geochemical equilibrium, triggering calcite dissolution. Different combinations of fluid chemistry and injection rate may therefore result in different evolution of porosity, permeability and dissolution patterns. To model the changes in porosity and permeability Kozeny-Carman equation K∝〖(∅)〗^n is used, where K is permeability and ∅ is porosity. The value of n is mostly based on experimental data or pore network models. In pore network models, this derivation is based on accuracy of relation used for conductivity and pore volume change. In fact, at a single pore scale, this relationship is the result of the pore shape development due to dissolution. We have prepared a new reactive transport model for a single pore which simulates the complex chemical reaction of carbonic-acid induced calcite dissolution and subsequent pore-geometry evolution at a single pore scale. We use COMSOL Multiphysics package 5.3 for the simulation. COMSOL utilizes the arbitary-Lagrangian Eulerian (ALE) method for the free-moving domain boundary. We examined the effect of flow rate on the evolution of single pore shape profiles due to calcite dissolution. We used three flow rates to cover diffusion dominated and advection-dominated transport regimes. The fluid in diffusion dominated flow (Pe number 0.037 and 0.37) becomes less reactive along the pore length and thus produced non-uniform pore shapes. However, for the advection-dominated flow (Pe number 3.75), the fast velocity of the fluid keeps the fluid relatively more reactive towards the end of the pore length, thus yielding uniform pore shape. Different pore shapes in terms of inlet opening vs overall pore opening will have an impact on the relation between changing volumes and conductivity. We have related the shape of pore with the Pe number which controls the transport regimes. For every Pe number, we have derived the relation between conductivity and porosity. These relations will be used in the pore network model to get the porosity and permeability variation.Keywords: single pore, reactive transport, calcite system, moving boundary
Procedia PDF Downloads 374428 Positive Interactions among Plants in Pinegroves over Quarzitic Sands
Authors: Enrique González Pendás, Vidal Pérez Hernández, Jorge Ferro Díaz, Nelson Careaga Pendás
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The investigation is carried out on the Protected Area of San Ubaldo, toward the interior of an open pinegrove with palm trees in a dry plainness of quar zitic sands, belonging to the Floristic Managed Reservation San Ubaldo-Sabanalamar, Guane, Pinar del Río, Cuba. This area is characterized by drastic seasonal variations, high temperatures and water evaporation, strong solar radiation, with sandy soils of almost pure quartz, which are very acid and poor in nutrients. The objective of the present work is to determine evidence of facilitation and its relationship with the structure and composition of plant communities in these peculiar ecosystems. For this study six lineal parallel transepts of 100 m are traced, in those, a general recording of the flora is carried out. To establish which plants act as nurses, is taken into account a height over 1 meter, canopy over 1.5 meter and the occurrence of several species under it. Covering was recorded using the line intercept method; the medium values of species richness for the taxa under nurses is compared with those that are located in open spaces among them. Then, it is determined which plants are better recruiter of other species (better nurses). An experiment is made to measure and compare some parameters in pine seedlings under the canopy of the Byrsonima crassifolia (L.) Kunth. and in open spaces, also the number of individuals is counted by species to calculate the frequency and total abundance in the study area. As a result, it is offered an up-to-date floristic list, a phylogenetic tree of the plant community showing a high phylodiversity, it is proven that the medium values of species richness and abundance of species under the nurses, is significantly superior to those occurring in open spaces. Furthermore, by means of phylogenetic trees it is shown that the species which cohabit under the nurses are not phylogenetically related. The former results are cited evidences of facilitation among plants, as well as it is one more time shown the importance of the nurse effect in preserving plant diversity on extreme environments.Keywords: facilitation, nurse plants, positive interactions, quarzitic sands
Procedia PDF Downloads 341427 Zinc Sorption by Six Agricultural Soils Amended with Municipal Biosolids
Authors: Antoine Karam, Lotfi Khiari, Bruno Breton, Alfred Jaouich
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Anthropogenic sources of zinc (Zn), including industrial emissions and effluents, Zn–rich fertilizer materials and pesticides containing Zn, can contribute to increasing the concentration of soluble Zn at levels toxic to plants in acid sandy soils. The application of municipal sewage sludge or biosolids (MBS) which contain metal immobilizing agents on coarse-textured soils could improve the metal sorption capacity of the low-CEC soils. The purpose of this experiment was to evaluate the sorption of Zn in surface samples (0-15 cm) of six Quebec (Canada) soils amended with MBS (pH 6.9) from Val d’Or (Quebec, Canada). Soil samples amended with increasing amounts (0 to 20%) of MBS were equilibrated with various amounts of Zn as ZnCl2 in 0.01 M CaCl2 for 48 hours at room temperature. Sorbed Zn was calculated from the difference between the initial and final Zn concentration in solution. Zn sorption data conformed to the linear form of Freundlich equation. The amount of sorbed Zn increased considerably with increasing MBS rate. Analysis of variance revealed a highly significant effect (p ≤ 0.001) of soil texture and MBS rate on the amount of sorbed Zn. The average values of the Zn-sorption capacity of MBS-amended coarse-textured soils were lower than those of MBS-amended fine textured soils. The two sandy soils (86-99% sand) amended with MBS retained 2- to 5-fold Zn than those without MBS (control). Significant Pearson correlation coefficients between the Zn sorption isotherm parameter, i.e. the Freundlich sorption isotherm (KF), and commonly measured physical and chemical entities were obtained. Among all the soil properties measured, soil pH gave the best significant correlation coefficients (p ≤ 0.001) for soils receiving 0, 5 and 10% MBS. Furthermore, KF values were positively correlated with soil clay content, exchangeable basic cations (Ca, Mg or K), CEC and clay content to CEC ratio. From these results, it can be concluded that (i) municipal biosolids provide sorption sites that have a strong affinity for Zn, (ii) both soil texture, especially clay content, and soil pH are the main factors controlling anthropogenic Zn sorption in the municipal biosolids-amended soils, and (iii) the effect of municipal biosolids on Zn sorption will be more pronounced for a sandy soil than for a clay soil.Keywords: metal, recycling, sewage sludge, trace element
Procedia PDF Downloads 284426 Improved Intracellular Protein Degradation System for Rapid Screening and Quantitative Study of Essential Fungal Proteins in Biopharmaceutical Development
Authors: Patarasuda Chaisupa, R. Clay Wright
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The selection of appropriate biomolecular targets is a crucial aspect of biopharmaceutical development. The Auxin-Inducible Degron Degradation (AID) technology has demonstrated remarkable potential in efficiently and rapidly degrading target proteins, thereby enabling the identification and acquisition of drug targets. The AID system also offers a viable method to deplete specific proteins, particularly in cases where the degradation pathway has not been exploited or when the adaptation of proteins, including the cell environment, occurs to compensate for the mutation or gene knockout. In this study, we have engineered an improved AID system tailored to deplete proteins of interest. This AID construct combines the auxin-responsive E3 ubiquitin ligase binding domain, AFB2, and the substrate degron, IAA17, fused to the target genes. Essential genes of fungi with the lowest percent amino acid similarity to human and plant orthologs, according to the Basic Local Alignment Search Tool (BLAST), were cloned into the AID construct in S. cerevisiae (AID-tagged strains) using a modular yeast cloning toolkit for multipart assembly and direct genetic modification. Each E3 ubiquitin ligase and IAA17 degron was fused to a fluorescence protein, allowing for real-time monitoring of protein levels in response to different auxin doses via cytometry. Our AID system exhibited high sensitivity, with an EC50 value of 0.040 µM (SE = 0.016) for AFB2, enabling the specific promotion of IAA17::target protein degradation. Furthermore, we demonstrate how this improved AID system enhances quantitative functional studies of various proteins in fungi. The advancements made in auxin-inducible protein degradation in this study offer a powerful approach to investigating critical target protein viability in fungi, screening protein targets for drugs, and regulating intracellular protein abundance, thus revolutionizing the study of protein function underlying a diverse range of biological processes.Keywords: synthetic biology, bioengineering, molecular biology, biotechnology
Procedia PDF Downloads 92425 Sustainable Development Change within Our Environs
Authors: Akinwale Adeyinka
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Critical natural resources such as clean ground water, fertile topsoil, and biodiversity are diminishing at an exponential rate, orders of magnitude above that at which they can be regenerated. Based on news on world population record, over 6 billion people on earth, and almost a quarter million added each day, the scale of human activity and environmental impact is unprecedented. Soaring human population growth over the past century has created a visible challenge to earth’s life support systems. In addition, the world faces an onslaught of other environmental threats including degenerated global climate change, global warming, intensified acid rain, stratospheric ozone depletion and health threatening pollution. Overpopulation and the use of deleterious technologies combine to increase the scale of human activities to a level that underlies these entire problems. These intensifying trends cannot continue indefinitely, hopefully, through increased understanding and valuation of ecosystems and their services, earth’s basic life-support system will be protected for the future.To say the fact, human civilization is now the dominant cause of change in the global environment. Now that our relationship to the earth has change so utterly, we have to see that change and understand its implication. These are actually 2 aspects to the challenges which we should believe. The first is to realize that our power to harm the earth can indeed have global and even permanent effects. Second is to realize that the only way to understand our new role as a co-architect of nature is to see ourselves as part of a complex system that does operate according to the same simple rules of cause and effect we are used to. So understanding the physical/biological dimension of earth system is an important precondition for making sensible policy to protect our environment. Because we believe Sustainable Development Is a matter of reconciling respect for the environment, social equity and economic profitability. Also, we strongly believe that environmental protection is naturally about reducing air and water pollution, but it also includes the improvement of the environmental performance of existing process. That is why we should always have it at the heart of our business that the environmental problem is not our effect on the environment so much as our relationship with the environment. We should always think of being environmental friendly in our operation.Keywords: Stratospheric ozone depletion ion , Climate Change, global warming, social equity and economic profitability
Procedia PDF Downloads 337424 Transformation of ectA Gene From Halomonas elongata in Tomato Plant
Authors: Narayan Moger, Divya B., Preethi Jambagi, Krishnaveni C. K., Apsana M. R., B. R. Patil, Basvaraj Bagewadi
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Salinity is one of the major threats to world food security. Considering the requirement for salt tolerant crop plants in the present study was undertaken to clone and transferred the salt tolerant ectA gene from marine ecosystem into agriculture crop system to impart salinity tolerance. Ectoine is the compatible solute which accumulates in the cell membrane, is known to be involved in salt tolerance activity in most of the Halophiles. The present situation is insisting to development of salt tolerant transgenic lines to combat abiotic stress. In this background, the investigation was conducted to develop transgenic tomato lines by cloning and transferring of ectA gene is an ectoine derivative capable of enzymatic action for the production of acetyl-diaminobutyric acid. The gene ectA is involved in maintaining the osmotic balance of plants. The PCR amplified ectA gene (579bp) was cloned into T/A cloning vector (pTZ57R/T). The construct pDBJ26 containing ectA gene was sequenced by using gene specific forward and reverse primers. Sequence was analyzed using BLAST algorithm to check similarity of ectA gene with other isolates. Highest homology of 99.66 per cent was found with ectA gene sequences of isolates Halomonas elongata with the available sequence information in NCBI database. The ectA gene was further sub cloned into pRI101-AN plant expression vector and transferred into E. coli DH5α for its maintenance. Further pDNM27 was mobilized into A. tumefaciens LBA4404 through tri-parental mating system. The recombinant Agrobacterium containing pDNM27 was transferred into tomato plants through In planta plant transformation method. Out of 300 seedlings, co-cultivated only twenty-seven plants were able to well establish under the greenhouse condition. Among twenty-seven transformants only twelve plants showed amplification with gene specific primers. Further work must be extended to evaluate the transformants at T1 and T2 generations for ectoine accumulation, salinity tolerance, plant growth and development and yield.Keywords: salinity, computable solutes, ectA, transgenic, in planta transformation
Procedia PDF Downloads 81423 A Single-Use Endoscopy System for Identification of Abnormalities in the Distal Oesophagus of Individuals with Chronic Reflux
Authors: Nafiseh Mirabdolhosseini, Jerry Zhou, Vincent Ho
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The dramatic global rise in acid reflux has also led to oesophageal adenocarcinoma (OAC) becoming the fastest-growing cancer in developed countries. While gastroscopy with biopsy is used to diagnose OAC patients, this labour-intensive and expensive process is not suitable for population screening. This study aims to design, develop, and implement a minimally invasive system to capture optical data of the distal oesophagus for rapid screening of potential abnormalities. To develop the system and understand user requirements, a user-centric approach was employed by utilising co-design strategies. Target users’ segments were identified, and 38 patients and 14 health providers were interviewed. Next, the technical requirements were developed based on consultations with the industry. A minimally invasive optical system was designed and developed considering patient comfort. This system consists of the sensing catheter, controller unit, and analysis program. Its procedure only takes 10 minutes to perform and does not require cleaning afterward since it has a single-use catheter. A prototype system was evaluated for safety and efficacy for both laboratory and clinical performance. This prototype performed successfully when submerged in simulated gastric fluid without showing evidence of erosion after 24 hours. The system effectively recorded a video of the mid-distal oesophagus of a healthy volunteer (34-year-old male). The recorded images were used to develop an automated program to identify abnormalities in the distal oesophagus. Further data from a larger clinical study will be used to train the automated program. This system allows for quick visual assessment of the lower oesophagus in primary care settings and can serve as a screening tool for oesophageal adenocarcinoma. In addition, this system is able to be coupled with 24hr ambulatory pH monitoring to better correlate oesophageal physiological changes with reflux symptoms. It also can provide additional information on lower oesophageal sphincter functions such as opening times and bolus retention.Keywords: endoscopy, MedTech, oesophageal adenocarcinoma, optical system, screening tool
Procedia PDF Downloads 88422 Black Soybeans Show Acute and Chronic Liver Protective Functions against CCl4 Induced Liver Damage
Authors: Cheng-Kuang Hsu, Chih-Hsiang Chang, Chi-Chih Wang
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Black soybeans contain high amount of antioxidants including polyphenols, anthocyanins and flavones. The protective function of black soybean against CCl4 (a strong oxidant) induced acute and chronic liver damage was investigated in vivo using SD rats or ICR mouse. The evaluation of CCl4 induced oxidative stress in the liver tissues included the measurements of the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST), the concentration of thiobarbituric acid reactive substances (TBARS), the activities of antioxidant enzymes (superoxide dismutase SOD, catalase, and glutathione peroxidase GPx), as well as the level of histological lesion in the liver tissues. For chronic experiment, a decoction at the concentration of 100 or 1000 mg/kg of body weight, produced by baking black soybean at 130°C for 5 min and followed by immerging in 100°C hot water for 20 min, showed the inhibitory effect against CCl4 induced liver damage in SD rats. Hot-water extract (80 °C for 30 min) from un-preheated black soybean at the concentration of 200 mg/kg of body weight could not reduce ALT and AST levels in CCl4 treated SD rats, but the hot-water extract from preheated black soybean did enhance antioxidant enzymes activities, decline ALT and AST levels. Specially, the hot-water extract from the seed cost of black soybean had the highest liver protective function since it can reduce vacuolization and necrosis in the liver tissues. For acute experiment, the hot-water extracts from black soybean and the seed coat, as well as pure cyanidin-3-glucoside (C3G) could reduce ALT and AST levels of CCl4 induced ICR mouse. The decoction and hot-water extract from the seed coat of black soybean had higher total polyphenols, anthocyanins and flavones contents than those extracts from whole black soybean. Such results agreed with high liver protective function in the decoction and hot-water from the seed coat of black soybean. Black soybean showed protective function only after preheating process (baking at 130°C for 5 to 10 min) because preheating treatment damaged the cell wall and made the extraction of the antioxidants more effectively.Keywords: black soybean, liver protective function, antioxidant, antioxidative stress
Procedia PDF Downloads 481421 Additional Method for the Purification of Lanthanide-Labeled Peptide Compounds Pre-Purified by Weak Cation Exchange Cartridge
Authors: K. Eryilmaz, G. Mercanoglu
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Aim: Purification of the final product, which is the last step in the synthesis of lanthanide-labeled peptide compounds, can be accomplished by different methods. Among these methods, the two most commonly used methods are C18 solid phase extraction (SPE) and weak cation exchanger cartridge elution. SPE C18 solid phase extraction method yields high purity final product, while elution from the weak cation exchanger cartridge is pH dependent and ineffective in removing colloidal impurities. The aim of this work is to develop an additional purification method for the lanthanide-labeled peptide compound in cases where the desired radionuclidic and radiochemical purity of the final product can not be achieved because of pH problem or colloidal impurity. Material and Methods: For colloidal impurity formation, 3 mL of water for injection (WFI) was added to 30 mCi of 177LuCl3 solution and allowed to stand for 1 day. 177Lu-DOTATATE was synthesized using EZAG ML-EAZY module (10 mCi/mL). After synthesis, the final product was mixed with the colloidal impurity solution (total volume:13 mL, total activity: 40 mCi). The resulting mixture was trapped in SPE-C18 cartridge. The cartridge was washed with 10 ml saline to remove impurities to the waste vial. The product trapped in the cartridge was eluted with 2 ml of 50% ethanol and collected to the final product vial via passing through a 0.22μm filter. The final product was diluted with 10 mL of saline. Radiochemical purity before and after purification was analysed by HPLC method. (column: ACE C18-100A. 3µm. 150 x 3.0mm, mobile phase: Water-Acetonitrile-Trifluoro acetic acid (75:25:1), flow rate: 0.6 mL/min). Results: UV and radioactivity detector results in HPLC analysis showed that colloidal impurities were completely removed from the 177Lu-DOTATATE/ colloidal impurity mixture by purification method. Conclusion: The improved purification method can be used as an additional method to remove impurities that may result from the lanthanide-peptide synthesis in which the weak cation exchange purification technique is used as the last step. The purification of the final product and the GMP compliance (the final aseptic filtration and the sterile disposable system components) are two major advantages.Keywords: lanthanide, peptide, labeling, purification, radionuclide, radiopharmaceutical, synthesis
Procedia PDF Downloads 163420 Structure-Reactivity Relationship of Some Rhᴵᴵᴵ and Osᴵᴵᴵ Complexes with N-Inert Ligands in Ionic Liquids
Authors: Jovana Bogojeski, Dusan Cocic, Nenad Jankovic, Angelina Petrovic
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Kinetically-inert transition metal complexes, such as Rh(III) and Os(III) complexes, attract increasing attention as leading scaffolds for the development of potential pharmacological agents due to their inertness and stability. Therefore, we have designed and fully characterized a few novel rhodium(III) and osmium(III) complexes with a tridentate nitrogen−donor chelate system. For some complexes, the crystal X-ray structure analysis was performed. Reactivity of the newly synthesized complexes towards small biomolecules, such as L-methionine (L-Met), guanosine-5’-monophosphate (5’-GMP), and glutathione (GSH) has been examined. Also, the reactivity of these complexes towards the DNA/RNA (Ribonucleic acid) duplexes was investigated. Obtained results show that the newly synthesized complexes exhibit good affinity towards the studied ligands. Results also show that the complexes react faster with the RNA duplex than with the DNA and that in the DNA duplex reaction is faster with 15mer GG than with the 22mer GG. The UV-Vis (Ultraviolet-visible spectroscopy) is absorption spectroscopy, and the EB (Ethidium bromide) displacement studies were used to examine the interaction of these complexes with CT-DNA and BSA (Bovine serum albumin). All studied complex showed good interaction ability with both the DNA and BSA. Furthermore, the DFT (Density-functional theory) calculation and docking studies were performed. The impact of the metal complex on the cytotoxicity was tested by MTT assay (a colorimetric assay for assessing cell metabolic activity) on HCT-116 lines (human colon cancer cell line). In addition, all these tests were repeated in the presence of several water-soluble biologically active ionic liquids. Attained results indicate that the ionic liquids increase the activity of the investigated complexes. All obtained results in this study imply that the introduction of different spectator ligand can be used to improve the reactivity of rhodium(III) and osmium(III) complexes. Finally, these results indicate that the examined complexes show reactivity characteristics needed for potential anti-tumor agents, with possible targets being both the DNA and proteins. Every new contribution in this field is highly warranted due to the current lack of clinically used Metallo-based alternatives to cisplatin.Keywords: biomolecules, ionic liquids, osmium(III), rhodium(III)
Procedia PDF Downloads 150419 Immobilizing Quorum Sensing Inhibitors on Biomaterial Surfaces
Authors: Aditi Taunk, George Iskander, Kitty Ka Kit Ho, Mark Willcox, Naresh Kumar
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Bacterial infections on biomaterial implants and medical devices accounts for 60-70% of all hospital acquired infections (HAIs). Treatment or removal of these infected devices results in high patient mortality and morbidity along with increased hospital expenses. In addition, with no effective strategies currently available and rapid development of antibacterial resistance has made device-related infections extremely difficult to treat. Therefore, in this project we have developed biomaterial surfaces using antibacterial compounds that inhibit biofilm formation by interfering with the bacterial communication mechanism known as quorum sensing (QS). This study focuses on covalent attachment of potent quorum sensing (QS) inhibiting compounds, halogenated furanones (FUs) and dihydropyrrol-2-ones (DHPs), onto glass surfaces. The FUs were attached by photoactivating the azide groups on the surface, and the acid functionalized DHPs were immobilized on amine surface via EDC/NHS coupling. The modified surfaces were tested in vitro against pathogenic organisms such as Staphylococcus aureus and Pseudomonas aeruginosa using confocal laser scanning microscopy (CLSM). Successful attachment of compounds on the substrates was confirmed by X-ray photoelectron spectroscopy (XPS) and contact angle measurements. The antibacterial efficacy was assessed, and significant reduction in bacterial adhesion and biofilm formation was observed on the FU and DHP coated surfaces. The activity of the coating was dependent upon the type of substituent present on the phenyl group of the DHP compound. For example, the ortho-fluorophenyl DHP (DHP-2) exhibited 79% reduction in bacterial adhesion against S. aureus and para-fluorophenyl DHP (DHP-3) exhibited 70% reduction against P. aeruginosa. The results were found to be comparable to DHP coated surfaces prepared in earlier study via Michael addition reaction. FUs and DHPs were able to retain their in vitro antibacterial efficacy after covalent attachment via azide chemistry. This approach is a promising strategy to develop efficient antibacterial biomaterials to reduce device related infections.Keywords: antibacterial biomaterials, biomedical device-related infections, quorum sensing, surface functionalization
Procedia PDF Downloads 268418 Efficiently Dispersed MnOx on Mesoporous 3D Cubic Support for Cyclohexene Epoxidation
Authors: G. Imran, A. Pandurangan
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Epoxides constitute important intermediates for the production of fine and bulk chemicals as well as valuable building blocks for the synthesis of a variety of bioactive molecules. Manganese oxides are used as selective catalyst for various redox type reactions and also effectively used in the field of catalytic disposal of pollutants. Non-toxic, cost efficient factor and more over existence of wide range of oxidation state (+2 to +7) makes catalyst more interesting for both academic research and industrial applications. However, the serious drawback lying is the lower surface area. Exceedingly dispersed manganese oxide grafted over mesoporous solid material KIT-6 through ALD (Atomic Layer Deposition) technique effectively catalyze cyclohexene with H2O2 (30% in water) to corresponding epoxides. Highly selective epoxide >99% with 55.7% conversion of cyclohexene was achieved using huge dispersed active sites of MnOx species containing catalysts. Various weight percent such as (1, 3, 5, 7 & 10 wt %) of manganese (II) acetylacetonate complex was employed as Mn source to post-graft via active silanol groups of KIT-6 and are designated as (Mn-G-KIT-6). XRD, N2 sorption, HR-TEM, DRS-UV-VIS, EPR and H2-TPR were employed for structural and textural properties. Immense Mn species of about 95% proportion on silica matrix obtained was evident from ICP-OES.The resulting materials exhibited Type IV adsorption isotherms indiacting mesopore in nanorange. Si-KIT-6 and Mn-G-KIT-6 materials exhibited surface area of 519-289 m2/g and with decrease in pore volume of 0.96-0.49 cm3/g with pore diameter ranging 7.9- 7.2 with increase in wt%. DRS-UV-VIS spectroscopy and EPR studies reveal that manganese coexists as Mn2+/3+ species as extra-framework sites and frame-work sites that result in dispersion on surface of silica matrix of KIT-6 and incorporated manganese sites with silanol groups along with small sized MnO cluster, evident from HR-TEM which increase with Mn content. Conventional production of epoxides by the intramolecular etherification of chlorohydrins formed by the reaction of alkenes with hypochlorous acid is the major drawbacks obtained recently. The most efficient synthesis of oxiranes (epoxides) is obtained by mesoporous catalysts (Mn-G-KIT-6) are presented here and discussed.Keywords: ALD, epoxidation, mesoporous, MnOx
Procedia PDF Downloads 184417 Comparison of Existing Predictor and Development of Computational Method for S- Palmitoylation Site Identification in Arabidopsis Thaliana
Authors: Ayesha Sanjana Kawser Parsha
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S-acylation is an irreversible bond in which cysteine residues are linked to fatty acids palmitate (74%) or stearate (22%), either at the COOH or NH2 terminal, via a thioester linkage. There are several experimental methods that can be used to identify the S-palmitoylation site; however, since they require a lot of time, computational methods are becoming increasingly necessary. There aren't many predictors, however, that can locate S- palmitoylation sites in Arabidopsis Thaliana with sufficient accuracy. This research is based on the importance of building a better prediction tool. To identify the type of machine learning algorithm that predicts this site more accurately for the experimental dataset, several prediction tools were examined in this research, including the GPS PALM 6.0, pCysMod, GPS LIPID 1.0, CSS PALM 4.0, and NBA PALM. These analyses were conducted by constructing the receiver operating characteristics plot and the area under the curve score. An AI-driven deep learning-based prediction tool has been developed utilizing the analysis and three sequence-based input data, such as the amino acid composition, binary encoding profile, and autocorrelation features. The model was developed using five layers, two activation functions, associated parameters, and hyperparameters. The model was built using various combinations of features, and after training and validation, it performed better when all the features were present while using the experimental dataset for 8 and 10-fold cross-validations. While testing the model with unseen and new data, such as the GPS PALM 6.0 plant and pCysMod mouse, the model performed better, and the area under the curve score was near 1. It can be demonstrated that this model outperforms the prior tools in predicting the S- palmitoylation site in the experimental data set by comparing the area under curve score of 10-fold cross-validation of the new model with the established tools' area under curve score with their respective training sets. The objective of this study is to develop a prediction tool for Arabidopsis Thaliana that is more accurate than current tools, as measured by the area under the curve score. Plant food production and immunological treatment targets can both be managed by utilizing this method to forecast S- palmitoylation sites.Keywords: S- palmitoylation, ROC PLOT, area under the curve, cross- validation score
Procedia PDF Downloads 77416 Bacterial Community Diversity in Soil under Two Tillage Systems
Authors: Dalia Ambrazaitienė, Monika Vilkienė, Danute Karcauskienė, Gintaras Siaudinis
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The soil is a complex ecosystem that is part of our biosphere. The ability of soil to provide ecosystem services is dependent on microbial diversity. T Tillage is one of the major factors that affect soil properties. The no-till systems or shallow ploughless tillage are opposite of traditional deep ploughing, no-tillage systems, for instance, increase soil organic matter by reducing mineralization rates and stimulating litter concentrations of the top soil layer, whereas deep ploughing increases the biological activity of arable soil layer and reduces the incidence of weeds. The role of soil organisms is central to soil processes. Although the number of microbial species in soil is still being debated, the metagenomic approach to estimate microbial diversity predicted about 2000 – 18 000 bacterial genomes in 1 g of soil. Despite the key role of bacteria in soil processes, there is still lack of information about the bacterial diversity of soils as affected by tillage practices. This study focused on metagenomic analysis of bacterial diversity in long-term experimental plots of Dystric Epihypogleyic Albeluvisols in western part of Lithuania. The experiment was set up in 2013 and had a split-plot design where the whole-plot treatments were laid out in a randomized design with three replicates. The whole-plot treatments consisted of two tillage methods - deep ploughing (22-25 cm) (DP), ploughless tillage (7-10 cm) (PT). Three subsamples (0-20 cm) were collected on October 22, 2015 for each of the three replicates. Subsamples from the DP and PT systems were pooled together wise to make two composition samples, one representing deep ploughing (DP) and the other ploughless tillage (PT). Genomic DNA from soil sample was extracted from approximately 200 mg field-moist soil by using the D6005 Fungal/Bacterial Miniprep set (Zymo Research®) following the manufacturer’s instructions. To determine bacterial diversity and community composition, we employed a culture – independent approach of high-throughput pyrosequencing of the 16S rRNA gene. Metagenomic sequencing was made with Illumina MiSeq platform in Base Clear Company. The microbial component of soil plays a crucial role in cycling of nutrients in biosphere. Our study was a preliminary attempt at observing bacterial diversity in soil under two common but contrasting tillage practices. The number of sequenced reads obtained for PT (161 917) was higher than DP (131 194). The 10 most abundant genus in soil sample were the same (Arthrobacter, Candidatus Saccharibacteria, Actinobacteria, Acidobacterium, Mycobacterium, Bacillus, Alphaproteobacteria, Longilinea, Gemmatimonas, Solirubrobacter), just the percent of community part was different. In DP the Arthrobacter and Acidobacterium consist respectively 8.4 % and 2.5%, meanwhile in PT just 5.8% and 2.1% of all community. The Nocardioides and Terrabacter were observed just in PT. This work was supported by the project VP1-3.1-ŠMM-01-V-03-001 NKPDOKT and National Science Program: The effect of long-term, different-intensity management of resources on the soils of different genesis and on other components of the agro-ecosystems [grant number SIT-9/2015] funded by the Research Council of Lithuania.Keywords: deep ploughing, metagenomics, ploughless tillage, soil community analysis
Procedia PDF Downloads 246415 Gossypol Extraction from Cotton Seed and Evaluation of Cotton Seed and Boll-cotton-pol Extract on Treatment of Cutaneous Leishmaniasis Resistant to Drugs
Authors: M. Mirmohammadi, S. Taghdisi, F. Anali
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Gossypol is a yellow anti-nutritional compound found in the cotton plant. This substance exists in the cottonseed and other parts of the cotton plant, such as bark, leaves, and stems. Chemically, gossypol is a very active polyphenolic aldehyde compound, and due to this polyphenolic structure, it has antioxidant and therapeutic properties. On the other hand, this compound, especially in free form, has many toxic effects, that its excessive consumption can be very dangerous for humans and animals. In this study, gossypol was extracted as a derivative compound of gossypol acetic acid from cottonseed using the n-hexane solvent with an efficiency of 0.84 ± 0.04, which compared to the Gossypol extracted from cottonseed oil with the same method (cold press) showed a significant difference with its efficiency of 1.14 ± 0.06. Therefore, it can be suggested to use cottonseed oil to extract this valuable compound. In the other part of this research, cottonseed extracts and cotton bolls extracts were obtained by two methods of soaking and Soxhlet with hydroalcoholic solvent taken with a ratio of (25:75), then by using extracts and corn starch powder, four herbal medicine code was created and after receiving the code of ethics (IR.SSU.REC.1398.136) the therapeutic effect of each one on the Cutaneous leishmaniasis resistant to drugs (caused by the leishmaniasis parasite) was investigated in real patients and its results was compared with the common drug glucantime (local ampoule) (n = 36). Statistical studies showed that the use of herbal medicines prepared with cottonseed extract and cotton bolls extract has a significant positive effect on the treatment of the disease’s wounds (p-value > 0.05) compared to the control group (only ethanol). Also, by comparing the average diameter of the wounds after a two-month treatment period, no significant difference was found between the use of ointment containing extracts and local glucantime ampoules (p-value < 0.05). Bolls extract extracted with the Soxhlet method showed the best therapeutic effects, although there was no significant difference between them (p-value < 0.05). Therefore, there is acceptable reliability to recommend this medicine for the treatment of Cutaneous leishmaniasis resistant to drugs without the side effects of the chemical drug glucantime and the pain of injecting the ampoule.Keywords: cottonseed oil, gossypol, cotton boll, cutaneous leishmaniasis
Procedia PDF Downloads 95414 Detecting Impact of Allowance Trading Behaviors on Distribution of NOx Emission Reductions under the Clean Air Interstate Rule
Authors: Yuanxiaoyue Yang
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Emissions trading, or ‘cap-and-trade', has been long promoted by economists as a more cost-effective pollution control approach than traditional performance standard approaches. While there is a large body of empirical evidence for the overall effectiveness of emissions trading, relatively little attention has been paid to other unintended consequences brought by emissions trading. One important consequence is that cap-and-trade could introduce the risk of creating high-level emission concentrations in areas where emitting facilities purchase a large number of emission allowances, which may cause an unequal distribution of environmental benefits. This study will contribute to the current environmental policy literature by linking trading activity with environmental injustice concerns and empirically analyzing the causal relationship between trading activity and emissions reduction under a cap-and-trade program for the first time. To investigate the potential environmental injustice concern in cap-and-trade, this paper uses a differences-in-differences (DID) with instrumental variable method to identify the causal effect of allowance trading behaviors on emission reduction levels under the clean air interstate rule (CAIR), a cap-and-trade program targeting on the power sector in the eastern US. The major data source is the facility-year level emissions and allowance transaction data collected from US EPA air market databases. While polluting facilities from CAIR are the treatment group under our DID identification, we use non-CAIR facilities from the Acid Rain Program - another NOx control program without a trading scheme – as the control group. To isolate the causal effects of trading behaviors on emissions reduction, we also use eligibility for CAIR participation as the instrumental variable. The DID results indicate that the CAIR program was able to reduce NOx emissions from affected facilities by about 10% more than facilities who did not participate in the CAIR program. Therefore, CAIR achieves excellent overall performance in emissions reduction. The IV regression results also indicate that compared with non-CAIR facilities, purchasing emission permits still decreases a CAIR participating facility’s emissions level significantly. This result implies that even buyers under the cap-and-trade program have achieved a great amount of emissions reduction. Therefore, we conclude little evidence of environmental injustice from the CAIR program.Keywords: air pollution, cap-and-trade, emissions trading, environmental justice
Procedia PDF Downloads 151413 Development of Antioxidant Rich Bakery Products by Applying Lysine and Maillard Reaction Products
Authors: Attila Kiss, Erzsébet Némedi, Zoltán Naár
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Due to the rapidly growing number of conscious customers in the recent years, more and more people look for products with positive physiological effects which may contribute to the preservation of their health. In response to these demands Food Science Research Institute of Budapest develops and introduces into the market new functional foods of guaranteed positive effect that contain bioactive agents. New, efficient technologies are also elaborated in order to preserve the maximum biological effect of the produced foods. The main objective of our work was the development of new functional biscuits fortified with physiologically beneficial ingredients. Bakery products constitute the base of the food nutrients’ pyramid, thus they might be regarded as foodstuffs of the largest consumed quantity. In addition to the well-known and certified physiological benefits of lysine, as an essential amino acid, a series of antioxidant type compounds is formed as a consequence of the occurring Maillard-reaction. Progress of the evoked Maillard-reaction was studied by applying diverse sugars (glucose, fructose, saccharose, isosugar) and lysine at several temperatures (120-170°C). Interval of thermal treatment was also varied (10-30 min). The composition and production technologies were tailored in order to reach the maximum of the possible biological benefits, so as to the highest antioxidant capacity in the biscuits. Out of the examined sugar components, theextent of the Maillard-reaction-driven transformation of glucose was the most pronounced at both applied temperatures. For the precise assessment of the antioxidant activity of the products FRAP and DPPH methods were adapted and optimised. To acquire an authentic and extensive mechanism of the occurring transformations, Maillard-reaction products were identified, and relevant reaction pathways were revealed. GC-MS and HPLC-MS techniques were applied for the analysis of the 60 generated MRPs and characterisation of actual transformation processes. 3 plausible major transformation routes might have been suggested based on the analytical result and the deductive sequence of possible occurring conversions between lysine and the sugars.Keywords: Maillard-reaction, lysine, antioxidant activity, GC-MS and HPLC-MS techniques
Procedia PDF Downloads 482412 Purple Spots on Historical Parchments: Confirming the Microbial Succession at the Basis of Biodeterioration
Authors: N. Perini, M. C. Thaller, F. Mercuri, S. Orlanducci, A. Rubechini, L. Migliore
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The preservation of cultural heritage is one of the major challenges of today’s society, because of the fundamental right of future generations to inherit it as the continuity with their historical and cultural identity. Parchments, consisting of a semi-solid matrix of collagen produced from animal skin (i.e., sheep or goats), are a significant part of the cultural heritage, being used as writing material for many centuries. Due to their animal origin, parchments easily undergo biodeterioration. The most common biological damage is characterized by isolated or coalescent purple spots that often leads to the detachment of the superficial layer and the loss of the written historical content of the document. Although many parchments with the same biodegradative features were analyzed, no common causative agent has been found so far. Very recently, a study was performed on a purple-damaged parchment roll dated back 1244 A.D, the A.A. Arm. I-XVIII 3328, belonging to the oldest collection of the Vatican Secret Archive (Fondo 'Archivum Arcis'), by comparing uncolored undamaged and purple damaged areas of the same document. As a whole, the study gave interesting results to hypothesize a model of biodeterioration, consisting of a microbial succession acting in two main phases: the first one, common to all the damaged parchments, is characterized by halophilic and halotolerant bacteria fostered by the salty environment within the parchment maybe induced by bringing of the hides; the second one, changing with the individual history of each parchment, determines the identity of its colonizers. The design of this model was pivotal to this study, performed by different labs of the Tor Vergata University (Rome, Italy), in collaboration with the Vatican Secret Archive. Three documents, belonging to a collection of dramatically damaged parchments archived as 'Faldone Patrizi A 19' (dated back XVII century A.D.), were analyzed through a multidisciplinary approach, including three updated technologies: (i) Next Generation Sequencing (NGS, Illumina) to describe the microbial communities colonizing the damaged and undamaged areas, (ii) RAMAN spectroscopy to analyze the purple pigments, (iii) Light Transmitted Analysis (LTA) to evaluate the kind and entity of the damage to native collagen. The metagenomic analysis obtained from NGS revealed DNA sequences belonging to Halobacterium salinarum mainly in the undamaged areas. RAMAN spectroscopy detected pigments within the purple spots, mainly bacteriorhodopsine/rhodopsin-like pigments, a purple transmembrane protein containing retinal and present in Halobacteria. The LTA technique revealed extremely damaged collagen structures in both damaged and undamaged areas of the parchments. In the light of these data, the study represents a first confirmation of the microbial succession model described above. The demonstration of this model is pivotal to start any possible new restoration strategy to bring back historical parchments to their original beauty, but also to open opportunities for intervention on a huge amount of documents.Keywords: biodeterioration, parchments, purple spots, ecological succession
Procedia PDF Downloads 171411 Gas Chromatography and Mass Spectrometry in Honey Fingerprinting: The Occurrence of 3,4-dihydro-3-oxoedulan and (E)-4-(r-1',t-2',c-4'-trihydroxy-3',6',6'-trimethylcyclohexyl)-but-3-en-2-one
Authors: Igor Jerkovic
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Owing to the attractive sensory properties and low odour thresholds, norisoprenoids (degraded carotenoid-like structures with 3,5,5-trimethylcyclohex-2-enoic unit) have been identified as aroma contributors in a number of different matrices. C₁₃-Norisoprenoids have been found among volatile organic compounds of various honey types as well as C₉//C₁₀-norisoprenoids or C₁₄/C₁₅-norisoprenoids. Besides degradation of abscisic acid (which produces, e.g., dehydrovomifoliol, vomifoliol, others), the cleavage of the C(9)=C(10) bond of other carotenoid precursors directly generates nonspecific C₁₃-norisoprenoids such as trans-β-damascenone, 3-hydroxy-trans-β-damascone, 3-oxo-α-ionol, 3-oxo-α-ionone, β-ionone found in various honey types. β-Damascenone and β-ionone smelling like honey, exhibit the lowest odour threshold values of all C₁₃-norisoprenoids. The presentation is targeted on two uncommon C₁₃-norisoprenoids in the honey flavor that could be used as specific or nonspecific chemical markers of the botanical origin. Namely, after screening of different honey types, the focus was directed on Centaruea cyanus L. and Allium ursinum L. honey. The samples were extracted by headspace solid-phase microextraction (HS-SPME) and ultrasonic solvent extraction (USE) and the extracts were analysed by gas chromatography and mass spectrometry (GC-MS). SPME fiber with divinylbenzene/carboxen/polydimethylsiloxane (DVB/CAR/PDMS) coating was applied for the research of C. cyanus honey headspace and predominant identified compound was 3,4-dihydro-3-oxoedulan (2,5,5,8a-tetramethyl-2,3,5,6,8,8a-hexahydro-7H-chromen-7-one also known as 2,3,5,6,8,8a-hexahydro-2,5,5,8a-tetramethyl-7H-1-benzo-pyran-7-one). The oxoedulan structure contains epoxide and it is more volatile in comparison with its hydroxylated precursors. This compound has not been found in other honey types and can be considered specific for C. cyanus honey. The dichloromethane extract of A. ursinum honey contained abundant (E)-4-(r-1',t-2',c-4'-trihydroxy-3',6',6'-trimethylcyclohexyl)-but-3-en-2-one that was previously isolated as dominant substance from the ether extracts of New Zealand thyme honey. Although a wide variety of degraded carotenoid-like substances have been identified from different honey types, this appears to be rare situation where 3,4-dihydro-3-oxoedulan and (E)-4-(r-1',t-2',c-4'-trihydroxy-3',6',6'-trimethylcyclohexyl)-but-3-en-2-one have been found that is of great importance for chemical fingerprinting and identification of the chemical biomarkers that can complement the pollen analysis as the major method for the honey classification.Keywords: 3, 4-dihydro-3-oxoedulan, (E)-4-(r-1', t-2', c-4'-trihydroxy-3', 6', 6'-trimethylcyclohexyl)-but-3-en-2-one, honey flavour, C₁₃-norisoprenoids
Procedia PDF Downloads 331410 Performance of a Lytic Bacteriophage Cocktail against Pseudomonas aeruginosa in Conditions That Simulate the Cystic Fibrosis Lung Environment
Authors: Isaac Martin, Abigail Lark, Sandra Morales, Eric W. Alton, Jane C. Davies
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Objectives: The cystic fibrosis (CF) lung is a unique microbiological niche, wherein harmful bacteria persist for many years despite antibiotic therapy. Pseudomonas aeruginosa (Pa), the major culprit leading to lung decline and increased mortality, thrives in the lungs of patients with CF due to several factors that have been linked with poor antibiotic performance. Our group is investigating alternative therapies including bacteriophage cocktails with which we have previously demonstrated efficacy against planktonic organisms. In this study, we explored the effects of a 4-phage cocktail on Pa grown in two different conditions, intended to mirror the CF lung: a) alongside standard antibiotic treatment in pre-formed biofilms (structures formed by Pa-secreted exopolysaccharides which provide both physical and cell division barriers to antimicrobials and host defenses and b) in an acidic environment postulated to be present in the CF airway due both to the primary defect in bicarbonate secretion and secondary effects of inflammation. Methods: 16 Pa strains from CF patients at the Royal Brompton Hospital were selected based on sensitivity to a) ceftazidime/ tobramycin and b) the phage cocktail in a conventional plaque assay. To assess efficacy of phage in biofilms, 96 well plates with Pa (5x10⁷ CFU/ ml) were incubated in static conditions, allowing adherent bacterial colonies to form for 24 hr. Ceftazidime and tobramycin (both at 2 × MIC) were added, +/- bacteriophage (4x10⁸ PFU/mL) for a further 24 hr. Cell viability and biomass were estimated using fluorescent resazurin and crystal violet assays, respectively. To evaluate the effect of pH, strains were grown planktonically in shaking 96 well plates at pH 6.0, 6.6, 7.0 and 7.5 with tobramycin or phage, at varying concentrations. Cell viability was quantified by fluorescent resazurin assay. Results: For the biofilm assay, treatment groups were compared with untreated controls and expressed as percent reduction in cell viability and biomass. Addition of the 4-phage cocktail resulted in a 1.3-fold reduction in cell viability and 1.7-fold reduction in biomass (p < 0.001) when compared to standard antibiotic treatment alone. Notably, there was a 50 ± 15% reduction in cell viability and 60 ± 12% reduction in biomass (95% CI) for the 4 biofilms demonstrating the most resistance to antibiotic treatment. 83% of strains tested (n=6) showed decreased bacterial killing by tobramycin at acidic pHs (p < 0.01). However, 25% of strains (n=12) showed improved phage killing at acidic pHs (p < 0.05), with none showing the pattern of reduced efficacy at acidic pH demonstrated by tobramycin. Conclusion: The 4-phage anti-Pa cocktail tested against Pa performs well in pre-formed biofilms and in acidic environments; two conditions intended to mimic the CF lung. To our knowledge, these are the first data looking at the effects of subtle pH changes on phage-mediated bacterial killing in the context of Pa infection. These findings contribute to a growing body of evidence supporting the use of nebulised lytic bacteriophage as a treatment in the context of lung infection.Keywords: biofilm, cystic fibrosis, pH, Pseudomonas aeruginosa, lytic bacteriophage
Procedia PDF Downloads 173409 Development of Biosensor Chip for Detection of Specific Antibodies to HSV-1
Authors: Zatovska T. V., Nesterova N. V., Baranova G. V., Zagorodnya S. D.
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In recent years, biosensor technologies based on the phenomenon of surface plasmon resonance (SPR) are becoming increasingly used in biology and medicine. Their application facilitates exploration in real time progress of binding of biomolecules and identification of agents that specifically interact with biologically active substances immobilized on the biosensor surface (biochips). Special attention is paid to the use of Biosensor analysis in determining the antibody-antigen interaction in the diagnostics of diseases caused by viruses and bacteria. According to WHO, the diseases that are caused by the herpes simplex virus (HSV), take second place (15.8%) after influenza as a cause of death from viral infections. Current diagnostics of HSV infection include PCR and ELISA assays. The latter allows determination the degree of immune response to viral infection and respective stages of its progress. In this regard, the searches for new and available diagnostic methods are very important. This work was aimed to develop Biosensor chip for detection of specific antibodies to HSV-1 in the human blood serum. The proteins of HSV1 (strain US) were used as antigens. The viral particles were accumulated in cell culture MDBK and purified by differential centrifugation in cesium chloride density gradient. Analysis of the HSV1 proteins was performed by polyacrylamide gel electrophoresis and ELISA. The protein concentration was measured using De Novix DS-11 spectrophotometer. The device for detection of antigen-antibody interactions was an optoelectronic two-channel spectrometer ‘Plasmon-6’, using the SPR phenomenon in the Krechman optical configuration. It was developed at the Lashkarev Institute of Semiconductor Physics of NASU. The used carrier was a glass plate covered with 45 nm gold film. Screening of human blood serums was performed using the test system ‘HSV-1 IgG ELISA’ (GenWay, USA). Development of Biosensor chip included optimization of conditions of viral antigen sorption and analysis steps. For immobilization of viral proteins 0.2% solution of Dextran 17, 200 (Sigma, USA) was used. Sorption of antigen took place at 4-8°C within 18-24 hours. After washing of chip, three times with citrate buffer (pH 5,0) 1% solution of BSA was applied to block the sites not occupied by viral antigen. It was found direct dependence between the amount of immobilized HSV1 antigen and SPR response. Using obtained biochips, panels of 25 positive and 10 negative for the content of antibodies to HSV-1 human sera were analyzed. The average value of SPR response was 185 a.s. for negative sera and from 312 to. 1264 a.s. for positive sera. It was shown that SPR data were agreed with ELISA results in 96% of samples proving the great potential of SPR in such researches. It was investigated the possibility of biochip regeneration and it was shown that application of 10 mM NaOH solution leads to rupture of intermolecular bonds. This allows reuse the chip several times. Thus, in this study biosensor chip for detection of specific antibodies to HSV1 was successfully developed expanding a range of diagnostic methods for this pathogen.Keywords: biochip, herpes virus, SPR
Procedia PDF Downloads 417408 Microbial Analysis of Street Vended Ready-to-Eat Meat around Thohoyandou Area, Vhembe District, Limpopo Province, RSA
Authors: Tshimangadzo Jeanette Raedani, Edgar Musie, Afsatou Traore
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Background: Street-vended meats, including chicken, pork, and beef, are popular in urban areas worldwide due to their convenience and affordability. However, these meats often pose a significant risk of foodborne diseases. The high water activity, protein content, and nearly neutral pH of meat create conditions conducive to the growth of pathogenic bacteria. Street foods, particularly meats, are frequently linked to outbreaks of foodborne illnesses due to potential contamination from improper handling and preparation. This study aimed to assess the microbial quality and safety of street-vended ready-to-eat meat sold in the Thohoyandou area. Method: The study involved collecting 168 samples of street-vended meat, split evenly between chicken (n=84) and beef (n=84), from various vendors around Thohoyandou. The samples were randomly selected and transported in sterile conditions to the Department of Food Microbiology at the University of Venda for analysis. Each 10-gram sample was cultured in selective media: MSA for Staphylococcus aureus, EMB for E. coli O157, XLD agar for Salmonella, and Sorbitol McConkey for Shigella. After initial culturing, the presumptive colonies were sub-cultured for purification and identified through Gram staining and biochemical tests, including Catalase, API 20E, Klingler Iron Agar Test, and Vitek 2 system. Antibiotic susceptibility was tested using agents such as Ampicillin, Chloramphenicol, Penicillin, Neomycin, Tetracycline, Streptomycin, and Amoxicillin. Molecular characterization was performed to identify E. coli pathotypes using multiplex PCR. Results: Out of 168 samples tested, 32 (19%) were positive for Staphylococcus spp., with the highest prevalence found in cooked chicken meat. The most common staphylococcus species identified were S. xylosus (13.2%) and S. saprophyticus (10.5%). E. coli was present in 29 (19.3%) of the samples, with the highest prevalence in fried chicken. Antibiotic susceptibility testing showed that 100% of E. coli isolates were resistant to Ampicillin, Tetracycline, and Penicillin, but 100% were susceptible to Neomycin. Staphylococcus spp. isolates were also 100% resistant to Ampicillin and 100% susceptible to Neomycin. The study detected a range of virulence genes in E. coli, with prevalence rates from 13.33% to 86.67%. The identified pathotypes included EPEC, EHEC, ETEC, EAEC, and EIEC, with many isolates showing mixed pathotypes. Conclusion: The study highlighted that the microbial quality and safety of street-vended meats in Thohoyandou are inadequate, rendering them unsafe for consumption. The presence of pathogenic microorganisms in both beef and chicken samples indicates significant risks associated with poor personal hygiene and food preparation practices. This underscores the need for improved monitoring and stricter food safety measures to prevent foodborne diseases and ensure consumer safety.Keywords: meat, microbial analysis, street vendors, E. coli
Procedia PDF Downloads 27407 Effects of the Coagulation Bath and Reduction Process on SO2 Adsorption Capacity of Graphene Oxide Fiber
Authors: Özge Alptoğa, Nuray Uçar, Nilgün Karatepe Yavuz, Ayşen Önen
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Sulfur dioxide (SO2) is a very toxic air pollutant gas and it causes the greenhouse effect, photochemical smog, and acid rain, which threaten human health severely. Thus, the capture of SO2 gas is very important for the environment. Graphene which is two-dimensional material has excellent mechanical, chemical, thermal properties, and many application areas such as energy storage devices, gas adsorption, sensing devices, and optical electronics. Further, graphene oxide (GO) is examined as a good adsorbent because of its important features such as functional groups (epoxy, carboxyl and hydroxyl) on the surface and layered structure. The SO2 adsorption properties of the fibers are usually investigated on carbon fibers. In this study, potential adsorption capacity of GO fibers was researched. GO dispersion was first obtained with Hummers’ method from graphite, and then GO fibers were obtained via wet spinning process. These fibers were converted into a disc shape, dried, and then subjected to SO2 gas adsorption test. The SO2 gas adsorption capacity of GO fiber discs was investigated in the fields of utilization of different coagulation baths and reduction by hydrazine hydrate. As coagulation baths, single and triple baths were used. In single bath, only ethanol and CaCl2 (calcium chloride) salt were added. In triple bath, each bath has a different concentration of water/ethanol and CaCl2 salt, and the disc obtained from triple bath has been called as reference disk. The fibers which were produced with single bath were flexible and rough, and the analyses show that they had higher SO2 adsorption capacity than triple bath fibers (reference disk). However, the reduction process did not increase the adsorption capacity, because the SEM images showed that the layers and uniform structure in the fiber form were damaged, and reduction decreased the functional groups which SO2 will be attached. Scanning Electron Microscopy (SEM), Fourier Transform Infrared Spectroscopy (FTIR), X-Ray Diffraction (XRD) analyzes were performed on the fibers and discs, and the effects on the results were interpreted. In the future applications of the study, it is aimed that subjects such as pH and additives will be examined.Keywords: coagulation bath, graphene oxide fiber, reduction, SO2 gas adsorption
Procedia PDF Downloads 360406 Electrospun Fibre Networks Loaded with Hydroxyapatite and Barium Titanate as Smart Scaffolds for Tissue Regeneration
Authors: C. Busuioc, I. Stancu, A. Nicoara, A. Zamfirescu, A. Evanghelidis
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The field of tissue engineering has expanded its potential due to the use of composite biomaterials belonging to increasingly complex systems, leading to bone substitutes with properties that are continuously improving to meet the patient's specific needs. Furthermore, the development of biomaterials based on ceramic and polymeric phases is an unlimited resource for future scientific research, with the final aim of restoring the original tissue functionality. Thus, in the first stage, composite scaffolds based on polycaprolactone (PCL) or polylactic acid (PLA) and inorganic powders were prepared by employing the electrospinning technique. The targeted powders were: commercial and laboratory synthesized hydroxyapatite (HAp), as well as barium titanate (BT). By controlling the concentration of the powder within the precursor solution, together with the processing parameters, different types of three-dimensional architectures were achieved. In the second stage, both the mineral powders and hybrid composites were investigated in terms of composition, crystalline structure, and microstructure so that to demonstrate their suitability for tissue engineering applications. Regarding the scaffolds, these were proven to be homogeneous on large areas and loaded with mineral particles in different proportions. The biological assays demonstrated that the addition of inorganic powders leads to modified responses in the presence of simulated body fluid (SBF) or cell cultures. Through SBF immersion, the biodegradability coupled with bioactivity were highlighted, with fiber fragmentation and surface degradation, as well as apatite layer formation within the testing period. Moreover, the final composites represent supports accepted by the cells, favoring implant integration. Concluding, the purposed fibrous materials based on bioresorbable polymers and mineral powders, produced by the electrospinning technique, represent candidates with considerable potential in the field of tissue engineering. Future improvements can be attained by optimizing the synthesis process or by simultaneous incorporation of multiple inorganic phases with well-defined biological action in order to fabricate multifunctional composites.Keywords: barium titanate, electrospinning, fibre networks, hydroxyapatite, smart scaffolds
Procedia PDF Downloads 111405 Impact of Long Term Application of Municipal Solid Waste on Physicochemical and Microbial Parameters and Heavy Metal Distribution in Soils in Accordance to Its Agricultural Uses
Authors: Rinku Dhanker, Suman Chaudhary, Tanvi Bhatia, Sneh Goyal
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Municipal Solid Waste (MSW), being a rich source of organic materials, can be used for agricultural applications as an important source of nutrients for soil and plants. This is also an alternative beneficial management practice for MSW generated in developing countries. In the present study, MSW treated soil samples from last four to six years at farmer’s field in Rohtak and Gurgaon states (Haryana, India) were collected. The samples were analyzed for all-important agricultural parameters and compared with the control untreated soil samples. The treated soil at farmer’s field showed increase in total N by 48 to 68%, P by 45.7 to 51.3%, and K by 60 to 67% compared to untreated soil samples. Application of sewage sludge at different sites led to increase in microbial biomass C by 60 to 68% compared to untreated soil. There was significant increase in total Cu, Cr, Ni, Fe, Pb, and Zn in all sewage sludge amended soil samples; however, concentration of all the metals were still below the current permitted (EU) limits. To study the adverse effect of heavy metals accumulation on various soil microbial activities, the sewage sludge samples (from wastewater treatment plant at Gurgaon) were artificially contaminated with heavy metal concentration above the EU limits. They were then applied to soil samples with different rates (0.5 to 4.0%) and incubated for 90 days under laboratory conditions. The samples were drawn at different intervals and analyzed for various parameters like pH, EC, total N, P, K, microbial biomass C, carbon mineralization, and diethylenetriaminepentaacetic acid (DTPA) exactable heavy metals. The results were compared to the uncontaminated sewage sludge. The increasing level of sewage sludge from 0.5 to 4% led to build of organic C and total N, P and K content at the early stages of incubation. But, organic C was decreased after 90 days because of decomposition of organic matter. Biomass production was significantly increased in both contaminated and uncontaminated sewage soil samples, but also led to slight increases in metal accumulation and their bioavailability in soil. The maximum metal concentrations were found in treatment with 4% of contaminated sewage sludge amendment.Keywords: heavy metal, municipal sewage sludge, sustainable agriculture, soil fertility and quality
Procedia PDF Downloads 286404 Investigations on Geopolymer Concrete Slabs
Authors: Akhila Jose
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The cement industry is one of the major contributors to the global warming due to the release of greenhouse gases. The primary binder in conventional concrete is Ordinary Portland cement (OPC) and billions of tons are produced annually all over the world. An alternative binding material to OPC is needed to reduce the environmental impact caused during the cement manufacturing process. Geopolymer concrete is an ideal material to substitute cement-based binder. Geopolymer is an inorganic alumino-silicate polymer. Geopolymer Concrete (GPC) is formed by the polymerization of aluminates and silicates formed by the reaction of solid aluminosilicates with alkali hydroxides or alkali silicates. Various Industrial bye- products like Fly Ash (FA), Rice Husk Ash (RHA), Ground granulated Blast Furnace Slag (GGBFS), Silica Fume (SF), Red mud (RM) etc. are rich in aluminates and silicates. Using by-products from other industries reduces the carbon dioxide emission and thus giving a sustainable way of reducing greenhouse gas emissions and also a way to dispose the huge wastes generated from the major industries like thermal plants, steel plants, etc. The earlier research about geopolymer were focused on heat cured fly ash based precast members and this limited its applications. The heat curing mechanism itself is highly cumbersome and costly even though they possess high compressive strength, low drying shrinkage and creep, and good resistance to sulphate and acid environments. GPC having comparable strength and durability characteristics of OPC were able to develop under ambient cured conditions is the solution making it a sustainable alternative in future. In this paper an attempt has been made to review and compare the feasibility of ambient cured GPC over heat cured geopolymer concrete with respect to strength and serviceability characteristics. The variation on the behavior of structural members is also reviewed to identify the research gaps for future development of ambient cured geopolymer concrete. The comparison and analysis of studies showed that GPC most importantly ambient cured type has a comparable behavior with respect to OPC based concrete in terms strength and durability criteria.Keywords: geopolymer concrete, oven heated, durability properties, mechanical properties
Procedia PDF Downloads 183403 Glyco-Biosensing as a Novel Tool for Prostate Cancer Early-Stage Diagnosis
Authors: Pavel Damborsky, Martina Zamorova, Jaroslav Katrlik
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Prostate cancer is annually the most common newly diagnosed cancer among men. An extensive number of evidence suggests that traditional serum Prostate-specific antigen (PSA) assay still suffers from a lack of sufficient specificity and sensitivity resulting in vast over-diagnosis and overtreatment. Thus, the early-stage detection of prostate cancer (PCa) plays undisputedly a critical role for successful treatment and improved quality of life. Over the last decade, particular altered glycans have been described that are associated with a range of chronic diseases, including cancer and inflammation. These glycans differences enable a distinction to be made between physiological and pathological state and suggest a valuable biosensing tool for diagnosis and follow-up purposes. Aberrant glycosylation is one of the major characteristics of disease progression. Consequently, the aim of this study was to develop a more reliable tool for early-stage PCa diagnosis employing lectins as glyco-recognition elements. Biosensor and biochip technology putting to use lectin-based glyco-profiling is one of the most promising strategies aimed at providing fast and efficient analysis of glycoproteins. The proof-of-concept experiments based on sandwich assay employing anti-PSA antibody and an aptamer as a capture molecules followed by lectin glycoprofiling were performed. We present a lectin-based biosensing assay for glycoprofiling of serum biomarker PSA using different biosensor and biochip platforms such as label-free surface plasmon resonance (SPR) and microarray with fluorescent label. The results suggest significant differences in interaction of particular lectins with PSA. The antibody-based assay is frequently associated with the sensitivity, reproducibility, and cross-reactivity issues. Aptamers provide remarkable advantages over antibodies due to the nucleic acid origin, stability and no glycosylation. All these data are further step for construction of highly selective, sensitive and reliable sensors for early-stage diagnosis. The experimental set-up also holds promise for the development of comparable assays with other glycosylated disease biomarkers.Keywords: biomarker, glycosylation, lectin, prostate cancer
Procedia PDF Downloads 406402 Biodiesel Production from Edible Oil Wastewater Sludge with Bioethanol Using Nano-Magnetic Catalysis
Authors: Wighens Ngoie Ilunga, Pamela J. Welz, Olewaseun O. Oyekola, Daniel Ikhu-Omoregbe
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Currently, most sludge from the wastewater treatment plants of edible oil factories is disposed to landfills, but landfill sites are finite and potential sources of environmental pollution. Production of biodiesel from wastewater sludge can contribute to energy production and waste minimization. However, conventional biodiesel production is energy and waste intensive. Generally, biodiesel is produced from the transesterification reaction of oils with alcohol (i.e., Methanol, ethanol) in the presence of a catalyst. Homogeneously catalysed transesterification is the conventional approach for large-scale production of biodiesel as reaction times are relatively short. Nevertheless, homogenous catalysis presents several challenges such as high probability of soap. The current study aimed to reuse wastewater sludge from the edible oil industry as a novel feedstock for both monounsaturated fats and bioethanol for the production of biodiesel. Preliminary results have shown that the fatty acid profile of the oilseed wastewater sludge is favourable for biodiesel production with 48% (w/w) monounsaturated fats and that the residue left after the extraction of fats from the sludge contains sufficient fermentable sugars after steam explosion followed by an enzymatic hydrolysis for the successful production of bioethanol [29% (w/w)] using a commercial strain of Saccharomyces cerevisiae. A novel nano-magnetic catalyst was synthesised from mineral processing alkaline tailings, mainly containing dolomite originating from cupriferous ores using a modified sol-gel. The catalyst elemental chemical compositions and structural properties were characterised by X-ray diffraction (XRD), scanning electron microscopy (SEM), Fourier transform infra-red (FTIR) and the BET for the surface area with 14.3 m²/g and 34.1 nm average pore diameter. The mass magnetization of the nano-magnetic catalyst was 170 emu/g. Both the catalytic properties and reusability of the catalyst were investigated. A maximum biodiesel yield of 78% was obtained, which dropped to 52% after the fourth transesterification reaction cycle. The proposed approach has the potential to reduce material costs, energy consumption and water usage associated with conventional biodiesel production technologies. It may also mitigate the impact of conventional biodiesel production on food and land security, while simultaneously reducing waste.Keywords: biodiesel, bioethanol, edible oil wastewater sludge, nano-magnetism
Procedia PDF Downloads 145