Search results for: bacterial gene clusters
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2971

Search results for: bacterial gene clusters

331 Mixotrophic Growth as a Tool for Increasing Polyhydroxyalkanoates (PHA) Production in Cyanobacteria

Authors: Zuzana Sedrlova, Eva Slaninova, Ines Fritz, Christina Daffert, Stanislav Obruca

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Cyanobacteria are ecologically extremely important phototrophic gram-negative bacteria capable of oxygenic photosynthesis. They synthesize many interesting metabolites such as glycogen, carotenoids, but the most interesting metabolites are polyhydroxyalkanoates (PHA). The main advantage of cyanobacteria is the fact they do not require costly organic substrate and, oppositely, cyanobacteria can fix CO₂. PHA serves primarily as a carbon and energy source and occurs in the form of intracellular granules in bacterial cells. It is possible, PHA helps cyanobacteria to survive stress conditions since increased PHA synthesis was observed during cultivation in stress conditions. PHA is microbial biopolymers that are biodegradable with similar properties as petrochemical synthetic plastics. Production of PHA by heterotrophic bacteria is expensive; for price reduction waste materials as input, materials are used. Positively, cyanobacteria principally do not require organic carbon substrate since they are capable of CO₂ fixation. In this work, we demonstrated that stress conditions lead to the highest obtained yields of PHA in cyanobacterial cultures. Two cyanobacterial cultures from genera Synechocystis were used in this work. Cultivations were performed either in Erlenmayer flask or in tube multicultivator. Multiple stressors were applied on cyanobacterial cultures, and stressors include PHA precursors. PHA precursors are chemical substances and some of them do not occur naturally in the environment. Cultivation with the same PHA precursors in the same concentration led to a 1,6x higher amount of PHA when a multicultivator was used. The highest amount of PHA reached 25 % of PHA in dry cyanobacterial biomass. Both strains are capable of co-polymer synthesis in the presence of their structural precursor. The composition of co-polymer differs in Synechocystis sp. PCC 6803 and Synechocystis salina CCALA 192. Synechocystis sp. PCC 6803 cultivated with γ-butyrolakton accumulated co-polymer of 3-hydroxybutyrate (3HB) and 4-hydroxybutyrate (4HB) the composition of the copolymer was 56 % of 4HB and 44 % of 3HB. The total amount of PHA, as well as yield of biomass, was lower than in control due to the toxic properties of γ-butyrolakton. Funding: This study was partly funded by the project GA19- 19-29651L of the Czech Science Foundation (GACR) and partly funded by the Austrian Science Fund (FWF), a project I 4082-B25. This work was supported by Brno, Ph.D. Talent – Funded by the Brno City Municipality.

Keywords: co-polymer, cyanobacteria, PHA, synechocystis

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330 The MicroRNA-2110 Suppressed Cell Proliferation and Migration Capacity in Hepatocellular Carcinoma Cells

Authors: Pelin Balcik Ercin

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Introduction: ZEB transcription factor family member ZEB2, has a role in epithelial to mesenchymal transition during development and metastasis. The altered circulating extracellular miRNAs expression is observed in diseases, and extracellular miRNAs have an important role in cancer cell microenvironment. In ChIP-Seq study, the expression of miR-2110 was found to be regulated by ZEB2. In this study, the effects of miR2110 on cell proliferation and migration of hepatocellular carcinoma (HCC) cells were examined. Material and Methods: SNU398 cells transfected with mimic miR2110 (20nM) (HMI0375, Sigma-Aldrich) and negative control miR (HMC0002, Sigma-Aldrich). MicroRNA isolation was accomplished with miRVANA isolation kit according to manufacturer instructions. cDNA synthesis was performed expression, respectively, and calibrated with Ct of controls. The real-time quantitative PCR (RT-qPCR) reaction was performed using the TaqMan Fast Advanced Master Mix (Thermo Sci.). Ct values of miR2110 were normalized to miR-186-5p and miR16-5p for the intracellular gene. Cell proliferation analysis was analyzed with the xCELLigence RTCA System. Wound healing assay was analyzed with the ImageJ program and relative fold change calculated. Results: The mimic-miR-2110 transfected SNU398 cells nearly nine-fold (log2) more miR-2110 expressed compared to negative control transfected cells. The mimic-miR-2110 transfected HCC cell proliferation significantly inhibited compared to the negative control cells. Furthermore, miR-2110-SNU398 cell migration capacity was relatively four-fold decreased compared to negative control-miR-SNU398 cells. Conclusion: Our results suggest the miR-2110 inhibited cell proliferation and also miR-2110 negatively affect cell migration compared to control groups in HCC cells. These data suggest the complexity of microRNA EMT transcription factors regulation. These initial results are pointed out the predictive biomarker capacity of miR-2110 in HCC.

Keywords: epithelial to mesenchymal transition, EMT, hepatocellular carcinoma cells, micro-RNA-2110, ZEB2

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329 Six Years Antimicrobial Resistance Trends among Bacterial Isolates in Amhara National Regional State, Ethiopia

Authors: Asrat Agalu Abejew

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Background: Antimicrobial resistance (AMR) is a silent tsunami and one of the top global threats to health care and public health. It is one of the common agendas globally and in Ethiopia. Emerging AMR will be a double burden to Ethiopia, which is facing a series of problems from infectious disease morbidity and mortality. In Ethiopia, although there are attempts to document AMR in healthcare institutions, comprehensive and all-inclusive analysis is still lacking. Thus, this study is aimed to determine trends in AMR from 2016-2021. Methods: A retrospective analysis of secondary data recorded in the Amhara Public Health Institute (APHI) from 2016 to 2021 G.C was conducted. Blood, Urine, Stool, Swabs, Discharge, body effusions, and other Microbiological specimens were collected from each study participants, and Bacteria identification and Resistance tests were done using the standard microbiologic procedure. Data was extracted from excel in August 2022, Trends in AMR were analyzed, and the results were described. In addition, the chi-square (X2) test and binary logistic regression were used, and a P. value < 0.05 was used to determine a significant association. Results: During 6 years period, there were 25143 culture and susceptibility tests. Overall, 265 (46.2%) bacteria were resistant to 2-4 antibiotics, 253 (44.2%) to 5-7 antibiotics, and 56 (9.7%) to >=8 antibiotics. The gram-negative bacteria were 166 (43.9%), 155 (41.5%), and 55 (14.6%) resistant to 2-4, 5-7, and ≥8 antibiotics, respectively, whereas 99(50.8%), 96(49.2% and 1 (0.5%) of gram-positive bacteria were resistant to 2-4, 5-7 and ≥8 antibiotics respectively. K. pneumonia 3783 (15.67%) and E. coli 3199 (13.25%) were the most commonly isolated bacteria, and the overall prevalence of AMR was 2605 (59.9%), where K. pneumonia 743 (80.24%), E. cloacae 196 (74.81%), A. baumannii 213 (66.56%) being the most common resistant bacteria for antibiotics tested. Except for a slight decline during 2020 (6469 (25.4%)), the overall trend of AMR is rising from year to year, with a peak in 2019 (8480 (33.7%)) and in 2021 (7508 (29.9%). If left un-intervened, the trend in AMR will increase by 78% of variation from the study period, as explained by the differences in years (R2=0.7799). Ampicillin, Augmentin, ciprofloxacin, cotrimoxazole, tetracycline, and Tobramycin were almost resistant to common bacteria they were tested. Conclusion: AMR is linearly increasing during the last 6 years. If left as it is without appropriate intervention after 15 years (2030 E.C), AMR will increase by 338.7%. A growing number of multi-drug resistant bacteria is an alarm to awake policymakers and those who do have the concern to intervene before it is too late. This calls for a periodic, integrated, and continuous system to determine the prevalence of AMR in commonly used antibiotics.

Keywords: AMR, trend, pattern, MDR

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328 Identification and Characterization of Antimicrobial Peptides Isolated from Entophytic Bacteria and Their Activity against Multidrug-Resistance Gram-Negative Bacteria in South Korea

Authors: Maryam Beiranvand

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Multi-drug resistance in various microorganisms has increased globally in many healthcare facilities. Less effective antimicrobial activity of drug therapies for infection control becomes trouble. Since 1980, no new type of antimicrobial drug has been identified, even though combinations of antibiotic drugs have been discovered almost every decade. Between 1981 and 2006, over 70% of novel pharmaceuticals and chemical agents came from natural sources. Microorganisms have yielded almost 22,000 natural compounds. The identification of antimicrobial components from endophytes bacteria could help overcome the threat posed by multi-drug resistant strains. The project aims to analyze and identify antimicrobial peptides isolated from entophytic bacteria and their activity against multidrug-resistant Gram-negative bacteria in South Korea. Endophytic Paenibacillus polymyxa. 4G3 isolated from the plant, Gynura procumbery exhibited considerable antimicrobial activity against Methicillin-resistant Staphylococcus aureus, and Escherichia coli. The Rapid Annotations using Subsystems Technology showed that the total size of the draft genome was 5,739,603bp, containing 5178 genes with 45.8% G+C content. Genome annotation using antiSMASH version 6.0.0 was performed, which predicted the most common types of non-ribosomal peptide synthetase (NRPS) and polyketide synthase (PKS). In this study, diethyl aminoethyl cellulose (DEAEC) resin was used as the first step in purifying for unknown peptides, and then the target protein was identified using hydrophilic and hydrophobic solutions, optimal pH, and step-by-step tests for antimicrobial activity. This crude was subjected to C18 chromatography and elution with 0, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, and 100% methanol, respectively. Only the fraction eluted with 20% -60% methanol demonstrated good antimicrobial activity against MDR E. coli. The concentration of the active fragment was measured by the Brad-ford test, and Protein A280 - Thermo Fisher Scientific at the end by examining the SDS PAGE Resolving Gel, 10% Acrylamide and purity were confirmed. Our study showed that, based on the combined results of the analysis and purification. P polymyxa. 4G3 has a high potential exists for producing novel functions of polymyxin E and bacitracin against bacterial pathogens.

Keywords: endophytic bacteria, antimicrobial activity, antimicrobial peptide, whole genome sequencing analysis, multi -drug resistance gram negative bacteria

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327 Photobiomodulation Activates WNT/β-catenin Signaling for Wound Healing in an in Vitro Diabetic Wound Model

Authors: Dimakatso B. Gumede, Nicolette N. Houreld

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Diabetic foot ulcers (DFUs) are a complication of diabetes mellitus (DM), a metabolic disease caused by insulin resistance or insufficiency, resulting in hyperglycaemia and low-grade chronic inflammation. Current therapies for treating DFUs include wound debridement, glycaemic control, and wound dressing. However, these therapies are moderately effective as there is a recurrence of these ulcers and an increased risk of lower limb amputations. Photobiomodulation (PBM), which is the application of non-invasive low-level light for wound healing at the spectrum of 660-1000 nm, has shown great promise in accelerating the healing of chronic wounds. However, its underlying mechanisms are not clearly defined. Studies have indicated that PBM induces wound healing via the activation of signaling pathways that are involved in tissue repair, such as the transforming growth factor-β (TGF-β). However, other signaling pathways, such as the WNT/β-catenin pathway, which is also critical for wound repair, have not been investigated. This study aimed to elucidate if PBM at 660 nm and a fluence of 5 J/cm² activates the WNT/β-catenin signaling pathway for wound healing in a diabetic cellular model. Human dermal fibroblasts (WS1) were continuously cultured high-glucose (26.5 mM D-glucose) environment to create a diabetic cellular model. A central scratch was created in the diabetic model to ‘wound’ the cells. The diabetic wounded (DW) cells were thereafter irradiated at 660 nm and a fluence of 5 J/cm². Cell migration, gene expression and protein assays were conducted at 24- and 48-h post-PBM. The results showed that PBM at 660 nm and a fluence of 5 J/cm² significantly increased cell migration in diabetic wounded cells at 24-h post-PBM. The expression of CTNNB1, ACTA2, COL1A1 and COL3A1 genes was also increased in DW cells post-PBM. Furthermore, there was increased cytoplasmic accumulation and nuclear localization of β-catenin at 24 h post-PBM. The findings in this study demonstrate that PBM activates the WNT/β-catenin signaling pathway by inducing the accumulation of β-catenin in diabetic wounded cells, leading to increased cell migration and expression of wound repair markers. These results thus indicate that PBM has the potential to improve wound healing in diabetic ulcers via activation of the WNT/β-catenin signaling pathway.

Keywords: wound healing, diabetic ulcers, photobiomodulation, WNT/β-catenin, signalling pathway

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326 Towards the Rapid Synthesis of High-Quality Monolayer Continuous Film of Graphene on High Surface Free Energy Existing Plasma Modified Cu Foil

Authors: Maddumage Don Sandeepa Lakshad Wimalananda, Jae-Kwan Kim, Ji-Myon Lee

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Graphene is an extraordinary 2D material that shows superior electrical, optical, and mechanical properties for the applications such as transparent contacts. Further, chemical vapor deposition (CVD) technique facilitates to synthesizing of large-area graphene, including transferability. The abstract is describing the use of high surface free energy (SFE) and nano-scale high-density surface kinks (rough) existing Cu foil for CVD graphene growth, which is an opposite approach to modern use of catalytic surfaces for high-quality graphene growth, but the controllable rough morphological nature opens new era to fast synthesis (less than the 50s with a short annealing process) of graphene as a continuous film over conventional longer process (30 min growth). The experiments were shown that high SFE condition and surface kinks on Cu(100) crystal plane existing Cu catalytic surface facilitated to synthesize graphene with high monolayer and continuous nature because it can influence the adsorption of C species with high concentration and which can be facilitated by faster nucleation and growth of graphene. The fast nucleation and growth are lowering the diffusion of C atoms to Cu-graphene interface, which is resulting in no or negligible formation of bilayer patches. High energy (500W) Ar plasma treatment (inductively Coupled plasma) was facilitated to form rough and high SFE existing (54.92 mJm-2) Cu foil. This surface was used to grow the graphene by using CVD technique at 1000C for 50s. The introduced kink-like high SFE existing point on Cu(100) crystal plane facilitated to faster nucleation of graphene with a high monolayer ratio (I2D/IG is 2.42) compared to another different kind of smooth morphological and low SFE existing Cu surfaces such as Smoother surface, which is prepared by the redeposit of Cu evaporating atoms during the annealing (RRMS is 13.3nm). Even high SFE condition was favorable to synthesize graphene with monolayer and continuous nature; It fails to maintain clean (surface contains amorphous C clusters) and defect-free condition (ID/IG is 0.46) because of high SFE of Cu foil at the graphene growth stage. A post annealing process was used to heal and overcome previously mentioned problems. Different CVD atmospheres such as CH4 and H2 were used, and it was observed that there is a negligible change in graphene nature (number of layers and continuous condition) but it was observed that there is a significant difference in graphene quality because the ID/IG ratio of the graphene was reduced to 0.21 after the post-annealing with H2 gas. Addition to the change of graphene defectiveness the FE-SEM images show there was a reduction of C cluster contamination of the surface. High SFE conditions are favorable to form graphene as a monolayer and continuous film, but it fails to provide defect-free graphene. Further, plasma modified high SFE existing surface can be used to synthesize graphene within 50s, and a post annealing process can be used to reduce the defectiveness.

Keywords: chemical vapor deposition, graphene, morphology, plasma, surface free energy

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325 Investigating the Urban Heat Island Phenomenon in A Desert City Aiming at Sustainable Buildings

Authors: Afifa Mohammed, Gloria Pignatta, Mattheos Santamouris, Evangelia Topriska

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Climate change is one of the global challenges that is exacerbated by the rapid growth of urbanizations. Urban Heat Island (UHI) phenomenon can be considered as an effect of the urbanization and it is responsible together with the Climate change of the overheating of urban cities and downtowns. The purpose of this paper is to quantify and perform analysis of UHI Intensity in Dubai, United Arab Emirates (UAE), through checking the relationship between the UHI and different meteorological parameters (e.g., temperature, winds speed, winds direction). Climate data were collected from three meteorological stations in Dubai (e.g., Dubai Airport - Station 1, Al-Maktoum Airport - Station 2 and Saih Al-Salem - Station 3) for a period of five years (e.g., 2014 – 2018) based upon hourly rates, and following clustering technique as one of the methodology tools of measurements. The collected data of each station were divided into six clusters upon the winds directions, either from the seaside or from the desert side, or from the coastal side which is in between both aforementioned winds sources, to investigate the relationship between temperature degrees and winds speed values through UHI measurements for Dubai Airport - Station 1 compared with the same of Al-Maktoum Airport - Station 2. In this case, the UHI value is determined by the temperature difference of both stations, where Station 1 is considered as located in an urban area and Station 2 is considered as located in a suburban area. The same UHI calculations has been applied for Al-Maktoum Airport - Station 2 and Saih Salem - Station 3 where Station 2 is considered as located in an urban area and Station 3 is considered as located in a suburban area. The performed analysis aims to investigate the relation between the two environmental parameters (e.g., Temperature and Winds Speed) and the Urban Heat Island (UHI) intensity when the wind comes from the seaside, from the desert, and the remaining directions. The analysis shows that the correlation between the temperatures with both UHI intensity (e.g., temperature difference between Dubai Airport - Station 1 and Saih Al-Salem - Station 3 and between Al-Maktoum Airport - Station 2 and Saih Al-Salem - Station 3 (through station 1 & 2) is strong and has a negative relationship when the wind is coming from the seaside comparing between the two stations 1 and 2, while the relationship is almost zero (no relation) when the wind is coming from the desert side. The relation is independent between the two parameters, e.g., temperature and UHI, on Station 2, during the same procedures, the correlation between the urban heat island UHI phenomenon and wind speed is weak for both stations when wind direction is coming from the seaside comparing the station 1 and 2, while it was found that there’s no relationship between urban heat island phenomenon and wind speed when wind direction is coming from desert side. The conclusion could be summarized saying that the wind coming from the seaside or from the desert side have a different effect on UHI, which is strongly affected by meteorological parameters. The output of this study will enable more determination of UHI phenomenon under desert climate, which will help to inform about the UHI phenomenon and intensity and extract recommendations in two main categories such as planning of new cities and designing of buildings.

Keywords: meteorological data, subtropical desert climate, urban climate, urban heat island (UHI)

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324 Role of P53 Codon 72 Polymorphism and miR-146a Rs2910164 Polymorphism in Breast Cancer

Authors: Marjan Moradi fard, Hossein Rassi, Masoud Houshmand

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Aim: Breast cancer is one of the most common cancers affecting the morbidity and mortality of Iranian women. This disease is a result of collective alterations of oncogenes and tumor suppressor genes. Studies have produced conflicting results concerning the role of p53 codon 72 polymorphism (G>C) and miR-146a rs2910164 polymorphism (G>C) on the risk of several cancers; therefore, a research was performed to estimate the association between the p53 codon 72 polymorphism and miR-146a rs2910164 polymorphism in breast cancer. Methods and Materials: A total of 45 archival breast cancer samples from Khatam hospital and 40 healthy samples were collected. Verification of each cancer reported in a relative was sought through the pathology reports of the hospital records. Then, DNA extracted from all samples by standard methods and p53 codon 72 polymorphism genotypes and miR-146a rs2910164 polymorphism genotypes were analyzed using multiplex PCR. The tubules, mitotic activity, necrosis, polymorphism and grade of breast cancer were staged by Nottingham histological grading and immunohistochemical staining of the sections from the paraffin wax embedded tissues for the expression of ER, PR and p53 was carried out using a standard method. Finally, data analysis was performed using the 7 version of the Epi Info(TM) 2012 software and test chi-square(x2) for trend. Results: Successful DNA extraction was assessed by PCR amplification of b-actin gene (99 bp). According to the results, p53 GG genotype and miR-146a rs2910164 CC genotype was significantly associated with increased risk of breast cancer in the study population. In this study, we established that tumors of p53 GG genotype and miR-146a rs2910164 CC genotype exhibited higher mitotic activity, higher polymorphism, lower necrosis, lower tubules, higher ER- and PR-negatives and lower TP53-positives than the other genotypes. Conclusion: The present study provided preliminary evidence that a p53 GG genotype may effect breast cancer risk in the study population, interacting synergistically with miR-146a rs2910164 CC genotype. Our results demonstrate that the testing of p53 codon 72 polymorphism genotypes and miR-146a rs2910164 polymorphism genotypes in combination with clinical parameters can serve as major risk factors in the early identification of breast cancers.

Keywords: breast cancer, miR-146a rs2910164 polymorphism, p53 codon 72 polymorphism, tumors, pathology reports

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323 Polyvinyl Alcohol Incorporated with Hibiscus Extract Microcapsules as Combined Active and Intelligent Composite Film for Meat Preservation

Authors: Ahmed F. Ghanem, Marwa I. Wahba, Asmaa N. El-Dein, Mohamed A. EL-Raey, Ghada E.A. Awad

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Numerous attempts are being performed in order to formulate suitable packaging materials for meat products. However, to the best of our knowledge, the incorporation of free hibiscus extract or its microcapsules in the pure polyvinyl alcohol (PVA) matrix as packaging materials for meats is seldom reported. Therefore, this study aims at protection of the aqueous crude extract of hibiscus flowers utilizing spry drying encapsulation technique. Fourier transform infrared (FTIR), scanning electron microscope (SEM), and zetasizer results confirmed the successful formation of assembled capsules via strong interactions, spherical rough microparticles, and ~ 235 nm of particle size, respectively. Also, the obtained microcapsules enjoy high thermal stability, unlike the free extract. Then, the obtained spray-dried particles were incorporated into the casting solution of the pure PVA film with a concentration 10 wt. %. The segregated free-standing composite films were investigated, compared to the neat matrix, with several characterization techniques such as FTIR, SEM, thermal gravimetric analysis (TGA), mechanical tester, contact angle, water vapor permeability, and oxygen transmission. The results demonstrated variations in the physicochemical properties of the PVA film after the inclusion of the free and the extract microcapsules. Moreover, biological studies emphasized the biocidal potential of the hybrid films against microorganisms contaminating the meat. Specifically, the microcapsules imparted not only antimicrobial but also antioxidant activities to PVA. Application of the prepared films on the real meat samples displayed low bacterial growth with a slight increase in the pH over the storage time up to 10 days at 4 oC which further proved the meat safety. Moreover, the colors of the films did not significantly changed except after 21 days indicating the spoilage of the meat samples. No doubt, the dual-functional of prepared composite films pave the way towards combined active/smart food packaging applications. This would play a vital role in the food hygiene, including also quality control and assurance.

Keywords: PVA, hibiscus, extraction, encapsulation, active packaging, smart and intelligent packaging, meat spoilage

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322 Comparison of the Isolation Rates and Characteristics of Salmonella Isolated from Antibiotic-Free and Conventional Chicken Meat Samples

Authors: Jin-Hyeong Park, Hong-Seok Kim, Jin-Hyeok Yim, Young-Ji Kim, Dong-Hyeon Kim, Jung-Whan Chon, Kun-Ho Seo

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Salmonella contamination in chicken samples can cause major health problems in humans. However, not only the effects of antibiotic treatment during growth but also the impacts of poultry slaughter line on the prevalence of Salmonella in final chicken meat sold to consumers are unknown. In this study, we compared the isolation rates and antimicrobial resistance of Salmonella between antibiotic-free, conventional, conventional Korean native retail chicken meat samples and clonal divergence of Salmonella isolates by multilocus sequence typing. In addition, the distribution of extended-spectrum β-lactamase (ESBL) genes in ESBL-producing Salmonella isolates was analyzed. A total of 72 retail chicken meat samples (n = 24 antibiotic-free broiler [AFB] chickens, n = 24 conventional broiler [CB] chickens, and n = 24 conventional Korean native [CK] chickens) were collected from local retail markets in Seoul, South Korea. The isolation rates of Salmonella were 66.6% in AFB chickens, 45.8% in CB chickens, and 25% in CK chickens. By analyzing the minimum inhibitory concentrations of β -lactam antibiotics with the disc-diffusion test, we found that 81.2% of Salmonella isolates from AFB chickens, 63.6% of isolates from CB chickens, and 50% of isolates from CK chickens were ESBL producers; all ESBL-positive isolates had the CTX-M-15 genotype. Interestingly, all ESBL-producing Salmonella were revealed as ST16 by multilocus sequence typing. In addition, all CTX-M-15-positive isolates had the genetic platform of blaCTX-M gene (IS26-ISEcp1-blaCTX-M-15-IS903), to the best of our knowledge, this is the first report in Salmonella around the world. The Salmonella ST33 strain (S. Hadar) isolated in this study has never been reported in South Korea. In conclusion, our findings showed that antibiotic-free retail chicken meat products were also largely contaminated with ESBL-producing Salmonella and that their ESBL genes and genetic platforms were the same as those isolated from conventional retail chicken meat products.

Keywords: antibiotic-free poultry, conventional poultry, multilocus sequence typing, extended-spectrum β-lactamase, antimicrobial resistance

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321 Establishing a Drug Discovery Platform to Progress Compounds into the Clinic

Authors: Sheraz Gul

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The requirements for progressing a compound to clinical trials is well established and relies on the results from in-vitro and in-vivo animal tests to indicate that it is likely to be safe and efficacious when testing in humans. The typical data package required will include demonstrating compound safety, toxicity, bioavailability, pharmacodynamics (potential effects of the compound on body systems) and pharmacokinetics (how the compound is potentially absorbed, distributed, metabolised and eliminated after dosing in humans). If the desired criteria are met and the compound meets the clinical Candidate criteria and is deemed worthy of further development, a submission to regulatory bodies such as the US Food & Drug Administration for an exploratory Investigational New Drug Study can be made. The purpose of this study is to collect data to establish that the compound will not expose humans to unreasonable risks when used in limited, early-stage clinical studies in patients or normal volunteer subjects (Phase I). These studies are also designed to determine the metabolism and pharmacologic actions of the drug in humans, the side effects associated with increasing doses, and, if possible, to gain early evidence on their effectiveness. In order to reach the above goals, we have developed a pre-clinical high throughput Absorption, Distribution, Metabolism and Excretion–Toxicity (ADME–Toxicity) panel of assays to identify compounds that are likely to meet the Lead and Candidate compound acceptance criteria. This panel includes solubility studies in a range of biological fluids, cell viability studies in cancer and primary cell-lines, mitochondrial toxicity, off-target effects (across the kinase, protease, histone deacetylase, phosphodiesterase and GPCR protein families), CYP450 inhibition (5 different CYP450 enzymes), CYP450 induction, cardio-toxicity (hERG) and gene-toxicity. This panel of assays has been applied to multiple compound series developed in a number of projects delivering Lead and clinical Candidates and examples from these will be presented.

Keywords: absorption, distribution, metabolism and excretion–toxicity , drug discovery, food and drug administration , pharmacodynamics

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320 Insights into the Annotated Genome Sequence of Defluviitoga tunisiensis L3 Isolated from a Thermophilic Rural Biogas Producing Plant

Authors: Irena Maus, Katharina Gabriella Cibis, Andreas Bremges, Yvonne Stolze, Geizecler Tomazetto, Daniel Wibberg, Helmut König, Alfred Pühler, Andreas Schlüter

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Within the agricultural sector, the production of biogas from organic substrates represents an economically attractive technology to generate bioenergy. Complex consortia of microorganisms are responsible for biomass decomposition and biogas production. Recently, species belonging to the phylum Thermotogae were detected in thermophilic biogas-production plants utilizing renewable primary products for biomethanation. To analyze adaptive genome features of representative Thermotogae strains, Defluviitoga tunisiensis L3 was isolated from a rural thermophilic biogas plant (54°C) and completely sequenced on an Illumina MiSeq system. Sequencing and assembly of the D. tunisiensis L3 genome yielded a circular chromosome with a size of 2,053,097 bp and a mean GC content of 31.38%. Functional annotation of the complete genome sequence revealed that the thermophilic strain L3 encodes several genes predicted to facilitate growth of this microorganism on arabinose, galactose, maltose, mannose, fructose, raffinose, ribose, cellobiose, lactose, xylose, xylan, lactate and mannitol. Acetate, hydrogen (H2) and carbon dioxide (CO2) are supposed to be end products of the fermentation process. The latter gene products are metabolites for methanogenic archaea, the key players in the final step of the anaerobic digestion process. To determine the degree of relatedness of dominant biogas community members within selected digester systems to D. tunisiensis L3, metagenome sequences from corresponding communities were mapped on the L3 genome. These fragment recruitments revealed that metagenome reads originating from a thermophilic biogas plant covered 95% of D. tunisiensis L3 genome sequence. In conclusion, availability of the D. tunisiensis L3 genome sequence and insights into its metabolic capabilities provide the basis for biotechnological exploitation of genome features involved in thermophilic fermentation processes utilizing renewable primary products.

Keywords: genome sequence, thermophilic biogas plant, Thermotogae, Defluviitoga tunisiensis

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319 The Effect of Slum Neighborhoods on Pregnancy Outcomes in Tanzania: Secondary Analysis of the 2015-2016 Tanzania Demographic and Health Survey Data

Authors: Luisa Windhagen, Atsumi Hirose, Alex Bottle

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Global urbanization has resulted in the expansion of slums, leaving over 10 million Tanzanians in urban poverty and at risk of poor health. Whilst rural residence has historically been associated with an increased risk of adverse pregnancy outcomes, recent studies found higher perinatal mortality rates in urban Tanzania. This study aims to understand to what extent slum neighborhoods may account for the spatial disparities seen in Tanzania. We generated a slum indicator based on UN-HABITAT criteria to identify slum clusters within the 2015-2016 Tanzania Demographic and Health Survey. Descriptive statistics, disaggregated by urban slum, urban non-slum, and rural areas, were produced. Simple and multivariable logistic regression examined the association between cluster residence type and neonatal mortality and stillbirth. For neonatal mortality, we additionally built a multilevel logistic regression model, adjusting for confounding and clustering. The neonatal mortality ratio was highest in slums (38.3 deaths per 1000 live births); the stillbirth rate was three times higher in slums (32.4 deaths per 1000 births) than in urban non-slums. Neonatal death was more likely to occur in slums than in urban non-slums (aOR=2.15, 95% CI=1.02-4.56) and rural areas (aOR=1.78, 95% CI=1.15-2.77). Odds of stillbirth were over five times higher among rural than urban non-slum residents (aOR=5.25, 95% CI=1.31-20.96). The results suggest that slums contribute to the urban disadvantage in Tanzanian neonatal health. Higher neonatal mortality in slums may be attributable to lack of education, lower socioeconomic status, poor healthcare access, and environmental factors, including indoor and outdoor air pollution and unsanitary conditions from inadequate housing. However, further research is required to ascertain specific causalities as well as significant associations between residence type and other pregnancy outcomes. The high neonatal mortality, stillbirth, and slum formation rates in Tanzania signify that considerable change is necessary to achieve international goals for health and human settlements. Disparities in access to adequate housing, safe water and sanitation, high standard antenatal, intrapartum, and neonatal care, and maternal education need to urgently be addressed. This study highlights the spatial neonatal mortality shift from rural settings to urban informal settlements in Tanzania. Importantly, other low- and middle-income countries experiencing overwhelming urbanization and slum expansion may also be at risk of a reversing trend in residential neonatal health differences.

Keywords: urban health, slum residence, neonatal mortality, stillbirth, global urbanisation

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318 Promoter Methylation of RASSF1A and MGMT Genes in Head and Neck Squamous Cell Carcinoma

Authors: Vitor Rafael Regiani, Carlos Henrique Viesi Do Nascimento Filho, Patricia Matos Biselli-Chicote, Claudia Aparecida Rainho, Luiz Sergio Raposo, José Victor Maniglia, Eny Maria Goloni-Bertollo, Erika Cristina Pavarino

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Promoter hypermethylation of tumor-related genes has been associated with prognosis in early-stage head-and-neck cancers, providing strong evidence that these hypermethylated genes are valuable biomarkers for prognostic evaluation. Hence, we selected the MGMT and RASSF1A genes to examine the methylation status in head and neck squamous cell carcinomas (HNSCC) samples matched with non-tumor tissues (tumor-surrounding tissues or peripheral blood samples). DNA methylation analysis was based on Methylation-Sensitive High Resolution Melting, and the methylation status was correlated with clinic-pathological characteristics of the patients. RASSF1A and MGMT promoter methylation was detected in 43.24% (16/37) and in 44.44% (16/36) of the tumors, respectively. RASSF1A and MGMT methylation was significantly more frequent in tumor tissue than non-tumor tissues, as well as, simultaneous methylation of RASSF1A and MGMT also was higher in tumor tissue than non-tumor tissues. In relation to anatomic site, larynx cancer presented significant methylation of MGMT gene compared to tumor-surrounding tissue. The frequency of RASSF1A and MGMT promoter methylated was higher in tumor tissues in relation to peripheral blood from the same patient. No association was found between methylation and the variables analyzed, including gender, age, smoking or alcohol drinking habits. Clinic-pathological characteristics also showed no association in the presence of methylation. The Kaplan–Meier's method showed no association of methylation and both disease-free and overall survival. In conclusion, the presence of epigenetic abnormalities in normal-appearing tissue corroborates the hypothesis of the ‘field cancerization', or it can reflect preneoplastic and/or preinvasive. Moreover, MGMT methylation may serve as an important laryngeal cancer biomarker because it showed significant difference between laryngeal cancer and surrounding tumor tissues.

Keywords: head and neck cancer, DNA methylation, MGMT promoter methylation, RASSF1A promoter methylation

Procedia PDF Downloads 308
317 Biological Control of Fusarium Crown and Root and Tomato (Solanum lycopersicum L.) Growth Promotion Using Endophytic Fungi from Withania somnifera L.

Authors: Nefzi Ahlem, Aydi Ben Abdallah Rania, Jabnoun-Khiareddine Hayfa, Ammar Nawaim, Mejda Daami-Remadi

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Fusarium Crown and Root Rot (FCRR) caused by Fusarium oxysporum f. sp. radicis-lycopersici (FORL) is a serious tomato (Solanum lycopersicum L.) disease in Tunisia. Its management is very difficult due to the long survival of its resting structures and to the luck of genetic resistance. In this work, we explored the wild Solanaceae species Withania somnifera, growing in the Tunisian Centre-East, as a potential source of biocontrol agents effective in FCRR suppression and tomato growth promotion. Seven fungal isolates were shown able to colonize tomato roots, crowns, and stems. Used as conidial suspensions or cell-free culture filtrates, all tested fungal treatments significantly enhanced tomato growth parameters by 21.5-90.3% over FORL-free control and by 27.6-93.5% over pathogen-inoculated control. All treatments significantly decreased the leaf and root damage index by 28.5-92.8 and the vascular browning extent 9.7-86.4% over FORL-inoculated and untreated control. The highest disease suppression ability (decrease by 86.4-92.8% in FCRR severity) over pathogen-inoculated control and by 81.3-88.8 over hymexazol-treated control) was expressed by I6 based treatments. This endophytic fungus was morphologically characterized and identified using rDNA sequencing gene as Fusarium sp. I6 (MG835371). This fungus was shown able to reduce FORL radial growth by 58.5–83.2% using its conidial suspension or cell-free culture filtrate. Fusarium sp. I6 showed chitinolytic, proteolytic and amylase activities. The current study clearly demonstrated that Fusarium sp. (I6) is a promising biocontrol candidate for suppressing FCRR severity and promoting tomato growth. Further investigations are required for elucidating its mechanism of action involved in disease suppression and plant growth promotion.

Keywords: antifungal activity, associated fungi, Fusarium oxysporum f. sp. radicis-lycopersici, Withania somnifera, tomato growth

Procedia PDF Downloads 138
316 A Rare Case of Synchronous Colon Adenocarcinoma

Authors: Mohamed Shafi Bin Mahboob Ali

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Introduction: Synchronous tumor is defined as the presence of more than one primary malignant lesion in the same patient at the indexed diagnosis. It is a rare occurrence, especially in the spectrum of colorectal cancer, which accounts for less than 4%. The underlying pathology of a synchronous tumor is thought to be due to a genomic factor, which is microsatellite instability (MIS) with the involvement of BRAF, KRAS, and the GSRM1 gene. There are no specific sites of occurrence for the synchronous colorectal tumor, but many studies have shown that a synchronous tumor has about 43% predominance in the ascending colon with rarity in the sigmoid colon. Case Report: We reported a case of a young lady in the middle of her 30's with no family history of colorectal cancer that was diagnosed with a synchronous adenocarcinoma at the descending colon and rectosigmoid region. The lady's presentation was quite perplexing as she presented to the district hospital initially with simple, uncomplicated hemorrhoids and constipation. She was then referred to our center for further management as she developed a 'football' sized right gluteal swelling with a complete intestinal obstruction and bilateral lower-limb paralysis. We performed a CT scan and biopsy of the lesion, which found that the tumor engulfed the sacrococcygeal region with more than one primary lesion in the colon as well as secondaries in the liver. The patient was operated on after a multidisciplinary meeting was held. Pelvic exenteration with tumor debulking and anterior resection were performed. Postoperatively, she was referred to the oncology team for chemotherapy. She had a tremendous recovery in eight months' time with a partial regain of her lower limb power. The patient is still under our follow-up with an improved quality of life post-intervention. Discussion: Synchronous colon cancer is rare, with an incidence of 2.4% to 12.4%. It has male predominance and is pathologically more advanced compared to a single colon lesion. Down staging the disease by means of chemoradiotherapy has shown to be effective in managing this tumor. It is seen commonly on the right colon, but in our case, we found it on the left colon and the rectosigmoid. Conclusion: Managing a synchronous colon tumor could be challenging to surgeons, especially in deciding the extent of resection and postoperative functional outcomes of the bowel; thus, individual treatment strategies are needed to tackle this pathology.

Keywords: synchronous, colon, tumor, adenocarcinoma

Procedia PDF Downloads 98
315 Antigenic Diversity of Theileria parva Isolates from Cattle and Buffalo at the Wildlife-Livestock Interface in Southern and Eastern Africa

Authors: Mukolwe D. Lubembe, Odongo O. David, Githaka Naftali, Kanduma Esther, Marinda Oosthuizen, Kgomotso P. Sibeko

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Theileriosis is a tick-borne disease of cattle caused by an apicomplexan protozoan parasite of the genus Theileria. In eastern and southern Africa, Theileria infections in cattle are caused by the species Theileria parva whose natural reservoir is the African buffalo (Syncerus caffer). Currently, East Coast Fever (ECF) caused by the cattle-derived Theileria parva is still a major problem in eastern Africa and some parts of southern Africa but not in South Africa following its eradication in the 1950s. However, Corridor disease (CD) caused by the buffalo-derived Theileria parva still remains a concern in South Africa. The diversity of Theileria parva in South Africa in comparison to other affected countries is poorly defined yet its known to be the survival strategy of this parasite. We assessed the antigenic diversity of Theileria parva isolates from Buffalo and cattle at the wildlife-livestock interface comparing samples from South Africa, Zimbabwe, Kenya, Tanzania, and Uganda. Antigenic epitopes of eight schizont antigen genes (Tp1, Tp3, Tp4, Tp5, Tp6, Tp7, Tp8 and Tp10) were amplified by PCR from genomic DNA extracted from blood samples collected from cattle and buffalo at the wildlife-livestock interface. Amplicons were purified and then sequenced on NGS platform. Full length open reading frames (ORFs) of two schizont antigen genes (Tp2 and Tp9) and one sporozoite antigen gene, p67 were also amplified from genomic DNA. Amplicons were then purified and cloned for sequencing. Analysis was based on sequence differences in the genes. Preliminary results show an extensively diverse population of Theileria parva circulating in buffalo and cattle populations at the wildlife-livestock interface. Diversity of the antigen genes contributes to the evasion of the immune system of the host by Theileria parva. This possess a concern in that, some of the Theileria parva populations may re-assort and become adapted to cattle to cause a form of theileriosis that is as fatal as ECF in areas where ECF was eradicated or is absent

Keywords: Theileria parva, east coast fever, corridor diseases, antigen genes, diversity

Procedia PDF Downloads 221
314 Assessment of Groundwater Quality in Karakulam Grama Panchayath in Thiruvananthapuram, Kerala State, South India

Authors: D. S. Jaya, G. P. Deepthi

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Groundwater is vital to the livelihoods and health of the majority of the people since it provides almost the entire water resource for domestic, agricultural and industrial uses. Groundwater quality comprises the physical, chemical, and bacteriological qualities. The present investigation was carried out to determine the physicochemical and bacteriological quality of the ground water sources in the residential areas of Karakulam Grama Panchayath in Thiruvananthapuram district, Kerala state in India. Karakulam is located in the eastern suburbs of Thiruvananthapuram city. The major drinking water source of the residents in the study area are wells. The present study aims to assess the portability and irrigational suitability of groundwater in the study area. The water samples were collected from randomly selected dug wells and bore wells in the study area during post monsoon and pre-monsoon seasons of the year 2014 after a preliminary field survey. The physical, chemical and bacteriological parameters of the water samples were analysed following standard procedures. The concentration of heavy metals (Cd, Pb, and Mn) in the acid digested water samples were determined by using an Atomic Absorption Spectrophotometer. The results showed that the pH of well water samples ranged from acidic to the alkaline level. In the majority of well water samples ( > 54%) the iron and magnesium content were found high in both the seasons studied, and the values were above the permissible limits of WHO drinking water quality standards. Bacteriological analyses showed that 63% of the wells were contaminated with total coliforms in both the seasons studied. Irrigational suitability of groundwater was assessed by determining the chemical indices like Sodium Percentage (%Na), Sodium Adsorption Ratio (SAR), Residual Sodium Carbonate (RSC), Permeability Index (PI), and the results indicate that the well water in the study area is good for irrigation purposes. Therefore, the study reveals the degradation of drinking water quality groundwater sources in Karakulam Grama Panchayath in Thiruvananthapuram District, Kerala in terms of its chemical and bacteriological characteristics and is not potable without proper treatment. In the study, more than 1/3rd of the wells tested were positive for total coliforms, and the bacterial contamination may pose threats to public health. The study recommends the need for periodic well water quality monitoring in the study area and to conduct awareness programs among the residents.

Keywords: bacteriological, groundwater, irrigational suitability, physicochemical, portability

Procedia PDF Downloads 254
313 A Novel Application of CORDYCEPIN (Cordycepssinensis Extract): Maintaining Stem Cell Pluripotency and Improving iPS Generation Efficiency

Authors: Shih-Ping Liu, Cheng-Hsuan Chang, Yu-Chuen Huang, Shih-Yin Chen, Woei-Cherng Shyu

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Embryonic stem cells (ES) and induced pluripotnet stem cells (iPS) are both pluripotent stem cells. For mouse stem cells culture technology, leukemia inhibitory factor (LIF) was used to maintain the pluripotency of stem cells in vitro. However, LIF is an expensive reagent. The goal of this study was to find out a pure compound extracted from Chinese herbal medicine that could maintain stem cells pluripotency to replace LIF and improve the iPS generation efficiency. From 20 candidates traditional Chinese medicine we found that Cordycepsmilitaris triggered the up-regulation of stem cells activating genes (Oct4 and Sox2) expression levels in MEF cells. Cordycepin, a major active component of Cordycepsmilitaris, also could up-regulate Oct4 and Sox2 gene expression. Furthermore, we used ES and iPS cells and treated them with different concentrations of Cordycepin (replaced LIF in the culture medium) to test whether it was useful to maintain the pluripotency. The results showed higher expression levels of several stem cells markers in 10 μM Cordycepin-treated ES and iPS cells compared to controls that did not contain LIF, including alkaline phosphatase, SSEA1, and Nanog. Embryonic body formation and differentiation confirmed that 10 μM Cordycepin-containing medium was capable to maintain stem cells pluripotency after four times passages. For mechanism analysis, microarray analysis indicated extracellular matrix and Jak/Stat signaling pathway as the top two deregulated pathways. In ECM pathway, we determined that the integrin αVβ5 expression levels and phosphorylated Src levels increased after Cordycepin treatment. In addition, the phosphorylated Jak2 and phosphorylated Sat3 protein levels were increased after Cordycepin treatment and suppressed with the Jak2 inhibitor, AG490. The expression of cytokines associated with Jak2/Stat3 signaling pathway were also up-regulated by Q-PCR and ELISA assay. Lastly, we used Oct4-GFP MEF cells to test iPS generation efficiency following Cordycepin treatment. We observed that 10 Μm Cordycepin significantly increased the iPS generation efficiency in day 21. In conclusion, we demonstrated Cordycepin could maintain the pluripotency of stem cells through both of ECM and Jak2/Stat3 signaling pathway and improved iPS generation efficiency.

Keywords: cordycepin, iPS cells, Jak2/Stat3 signaling pathway, molecular biology

Procedia PDF Downloads 425
312 Double Functionalization of Magnetic Colloids with Electroactive Molecules and Antibody for Platelet Detection and Separation

Authors: Feixiong Chen, Naoufel Haddour, Marie Frenea-Robin, Yves MéRieux, Yann Chevolot, Virginie Monnier

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Neonatal thrombopenia occurs when the mother generates antibodies against her baby’s platelet antigens. It is particularly critical for newborns because it can cause coagulation troubles leading to intracranial hemorrhage. In this case, diagnosis must be done quickly to make platelets transfusion immediately after birth. Before transfusion, platelet antigens must be tested carefully to avoid rejection. The majority of thrombopenia (95 %) are caused by antibodies directed against Human Platelet Antigen 1a (HPA-1a) or 5b (HPA-5b). The common method for antigen platelets detection is polymerase chain reaction allowing for identification of gene sequence. However, it is expensive, time-consuming and requires significant blood volume which is not suitable for newborns. We propose to develop a point-of-care device based on double functionalized magnetic colloids with 1) antibodies specific to antigen platelets and 2) highly sensitive electroactive molecules in order to be detected by an electrochemical microsensor. These magnetic colloids will be used first to isolate platelets from other blood components, then to capture specifically platelets bearing HPA-1a and HPA-5b antigens and finally to attract them close to sensor working electrode for improved electrochemical signal. The expected advantages are an assay time lower than 20 min starting from blood volume smaller than 100 µL. Our functionalization procedure based on amine dendrimers and NHS-ester modification of initial carboxyl colloids will be presented. Functionalization efficiency was evaluated by colorimetric titration of surface chemical groups, zeta potential measurements, infrared spectroscopy, fluorescence scanning and cyclic voltammetry. Our results showed that electroactive molecules and antibodies can be immobilized successfully onto magnetic colloids. Application of a magnetic field onto working electrode increased the detected electrochemical signal. Magnetic colloids were able to capture specific purified antigens extracted from platelets.

Keywords: Magnetic Nanoparticles , Electroactive Molecules, Antibody, Platelet

Procedia PDF Downloads 257
311 The Data Quality Model for the IoT based Real-time Water Quality Monitoring Sensors

Authors: Rabbia Idrees, Ananda Maiti, Saurabh Garg, Muhammad Bilal Amin

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IoT devices are the basic building blocks of IoT network that generate enormous volume of real-time and high-speed data to help organizations and companies to take intelligent decisions. To integrate this enormous data from multisource and transfer it to the appropriate client is the fundamental of IoT development. The handling of this huge quantity of devices along with the huge volume of data is very challenging. The IoT devices are battery-powered and resource-constrained and to provide energy efficient communication, these IoT devices go sleep or online/wakeup periodically and a-periodically depending on the traffic loads to reduce energy consumption. Sometime these devices get disconnected due to device battery depletion. If the node is not available in the network, then the IoT network provides incomplete, missing, and inaccurate data. Moreover, many IoT applications, like vehicle tracking and patient tracking require the IoT devices to be mobile. Due to this mobility, If the distance of the device from the sink node become greater than required, the connection is lost. Due to this disconnection other devices join the network for replacing the broken-down and left devices. This make IoT devices dynamic in nature which brings uncertainty and unreliability in the IoT network and hence produce bad quality of data. Due to this dynamic nature of IoT devices we do not know the actual reason of abnormal data. If data are of poor-quality decisions are likely to be unsound. It is highly important to process data and estimate data quality before bringing it to use in IoT applications. In the past many researchers tried to estimate data quality and provided several Machine Learning (ML), stochastic and statistical methods to perform analysis on stored data in the data processing layer, without focusing the challenges and issues arises from the dynamic nature of IoT devices and how it is impacting data quality. A comprehensive review on determining the impact of dynamic nature of IoT devices on data quality is done in this research and presented a data quality model that can deal with this challenge and produce good quality of data. This research presents the data quality model for the sensors monitoring water quality. DBSCAN clustering and weather sensors are used in this research to make data quality model for the sensors monitoring water quality. An extensive study has been done in this research on finding the relationship between the data of weather sensors and sensors monitoring water quality of the lakes and beaches. The detailed theoretical analysis has been presented in this research mentioning correlation between independent data streams of the two sets of sensors. With the help of the analysis and DBSCAN, a data quality model is prepared. This model encompasses five dimensions of data quality: outliers’ detection and removal, completeness, patterns of missing values and checks the accuracy of the data with the help of cluster’s position. At the end, the statistical analysis has been done on the clusters formed as the result of DBSCAN, and consistency is evaluated through Coefficient of Variation (CoV).

Keywords: clustering, data quality, DBSCAN, and Internet of things (IoT)

Procedia PDF Downloads 129
310 Biosynthesized Selenium Nanoparticles to Rescue Coccidiosis-mediated Oxidative Stress, Apoptosis and Inflammation in the Jejunum Of Mice

Authors: Esam Mohammed Al-shaebi

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One of the most crucial approaches for treating human diseases, particularly parasite infections, is nanomedicine. One of the most significant protozoan diseases that impact farm and domestic animals is coccidiosis. While, amprolium is one of the traditional anticoccidial medication, the advent of drug-resistant strains of Eimeria necessitates the development of novel treatments. The goal of the current investigation was to determine whether biosynthesized selenium nanoparticles (Bio-SeNPs) using Azadirachta indica leaves extract might treat mice with Eimeria papillata infection in the jejunal tissue. Five groups of seven mice each were used, as follows: Group 1: Non-infected-non-treated (negative control). Group 2: Non-infected treated group with Bio-SeNPs (0.5 mg/kg of body weight). Groups 3-5 were orally inoculated with 1×103 sporulated oocysts of E. papillata. Group 3: Infected-non-treated (positive control). Group 4: Infected and treated group with Bio-SeNPs (0.5 mg/kg). Group 5: Infected and treated group with the Amprolium. Groups 4 and 5 daily received oral administration (for 5 days) of Bio-SeNPs and anticoccidial medication, respectively, after infection. Bio-SeNPs caused a considerable reduction in oocyst output in mice feces (97.21%). This was also accompanied by a significant reduction in the number of developmental parasitic stages in the jejunal tissues. Glutathione reduced (GSH), glutathione peroxidase (GPx), and superoxide dismutase (SOD) levels were dramatically reduced by the Eimeria parasite, whereas, nitric oxide (NO) and malonaldehyde (MDA) levels were markedly elevated. The amount of goblet cells and MUC2 gene expression were used as apoptotic indicators, and both were considerably downregulated by infection. However, infection markedly increased the expression of inflammatory cytokines (IL-6 and TNF-α) and the apoptotic genes (Caspase-3 and BCL2). Bio-SeNPs were administrated to mice to drastically lower body weight, oxidative stress, and inflammatory and apoptotic indicators in the jejunal tissue. Our research thus showed the involvement of Bio-SeNPs in protecting mice with E. papillata infections against jejunal damage.

Keywords: coccidiosis, nanoparticles, azadirachta indica, oxidative stress

Procedia PDF Downloads 82
309 Functionalized Spherical Aluminosilicates in Biomedically Grade Composites

Authors: Damian Stanislaw Nakonieczny, Grazyna Simha Martynkova, Marianna Hundakova, G. Kratosová, Karla Cech Barabaszova

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The main aim of the research was to functionalize the surface of spherical aluminum silicates in the form of so-called cenospheres. Cenospheres are light ceramic particles with a density between 0.45 and 0.85 kgm-3 hat can be obtained as a result of separation from fly ash from coal combustion. However, their occurrence is limited to about 1% by weight of dry ash mainly derived from anthracite. Hence they are very rare and desirable material. Cenospheres are characterized by complete chemical inertness. Mohs hardness in range of 6 and completely smooth surface. Main idea was to prepare the surface by chemical etching, among others hydrofluoric acid (HF) and hydrogen peroxide, caro acid, silanization using (3-aminopropyl) triethoxysilane (APTES) and tetraethyl orthosilicate (TEOS) to obtain the maximum development and functionalization of the surface to improve chemical and mechanical connection with biomedically used polymers, i.e., polyacrylic methacrylate (PMMA) and polyetheretherketone (PEEK). These polymers are used medically mainly as a material for fixed and removable dental prostheses and PEEK spinal implants. The problem with their use is the decrease in mechanical properties over time and bacterial infections fungal during implantation and use of dentures. Hence, the use of a ceramic filler that will significantly improve the mechanical properties, improve the fluidity of the polymer during shape formation, and in the future, will be able to support bacteriostatic substances such as silver and zinc ions seem promising. In order to evaluate our laboratory work, several instrumental studies were performed: chemical composition and morphology with scanning electron microscopy with Energy-Dispersive X-Ray Probe (SEM/EDX), determination of characteristic functional groups of Fourier Transform Infrared Spectroscopy (FTIR), phase composition of X-ray Diffraction (XRD) and thermal analysis of Thermo Gravimetric Analysis/differentia thermal analysis (TGA/DTA), as well as assessment of isotherm of adsorption with Brunauer-Emmett-Teller (BET) surface development. The surface was evaluated for the future application of additional bacteria and static fungus layers. Based on the experimental work, it was found that orated methods can be suitable for the functionalization of the surface of cenosphere ceramics, and in the future it can be suitable as a bacteriostatic filler for biomedical polymers, i.e., PEEK or PMMA.

Keywords: bioceramics, composites, functionalization, surface development

Procedia PDF Downloads 109
308 Religiosity and Involvement in Purchasing Convenience Foods: Using Two-Step Cluster Analysis to Identify Heterogenous Muslim Consumers in the UK

Authors: Aisha Ijaz

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The paper focuses on the impact of Muslim religiosity on convenience food purchases and involvement experienced in a non-Muslim culture. There is a scarcity of research on the purchasing patterns of Muslim diaspora communities residing in risk societies, particularly in contexts where there is an increasing inclination toward industrialized food items alongside a renewed interest in the concept of natural foods. The United Kingdom serves as an appropriate setting for this study due to the increasing Muslim population in the country, paralleled by the expanding Halal Food Market. A multi-dimensional framework is proposed, testing for five forms of involvement, specifically Purchase Decision Involvement, Product Involvement, Behavioural Involvement, Intrinsic Risk and Extrinsic Risk. Quantitative cross-sectional consumer data were collected through a face-to-face survey contact method with 141 Muslims during the summer of 2020 in Liverpool located in the Northwest of England. proportion formula was utilitsed, and the population of interest was stratified by gender and age before recruitment took place through local mosques and community centers. Six input variables were used (intrinsic religiosity and involvement dimensions), dividing the sample into 4 clusters using the Two-Step Cluster Analysis procedure in SPSS. Nuanced variances were observed in the type of involvement experienced by religiosity group, which influences behaviour when purchasing convenience food. Four distinct market segments were identified: highly religious ego-involving (39.7%), less religious active (26.2%), highly religious unaware (16.3%), less religious concerned (17.7%). These segments differ significantly with respects to their involvement, behavioural variables (place of purchase and information sources used), socio-cultural (acculturation and social class), and individual characteristics. Choosing the appropriate convenience food is centrally related to the value system of highly religious ego-involving first-generation Muslims, which explains their preference for shopping at ethnic food stores. Less religious active consumers are older and highly alert in information processing to make the optimal food choice, relying heavily on product label sources. Highly religious unaware Muslims are less dietary acculturated to the UK diet and tend to rely on digital and expert advice sources. The less-religious concerned segment, who are typified by younger age and third generation, are engaged with the purchase process because they are worried about making unsuitable food choices. Research implications are outlined and potential avenues for further explorations are identified.

Keywords: consumer behaviour, consumption, convenience food, religion, muslims, UK

Procedia PDF Downloads 46
307 Microbiological Examination and Antimicrobial Susceptibility of Microorganisms Isolated from Salt Mining Site in Ebonyi State

Authors: Anyimc, C. J. Aneke, J. O. Orji, O. Nworie, U. C. C. Egbule

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The microbial examination and antimicrobial susceptibility profile of microorganism isolated from the salt mining site in Ebonyi state were evaluated in the present study using a standard microbiological technique. A total of 300 samples were randomly collected in three sample groups (A, B, and C) of 100 each. Isolation, Identification and characterization of organization present on the soil samples were determined by culturing, gram-staining and biochemical technique. The result showed the following organisms were isolated with their frequency as follow: Bacillus species (37.3%) and Staphylococcus species(23.5%) had the highest frequency in the whole Sample group A and B while Klebsiella specie (15.7%), Pseudomonas species(13.7%), and Erwinia species (9.8%) had the least. Rhizopus species (42.0%) and Aspergillus species (26.0%) were the highest fungi isolated, followed by Penicillum species (20.0%) while Mucor species (4.0%), and Fusarium species (8.0%) recorded the least. Sample group C showed high microbial population of all the microbial isolates when compared to sample group A and B. Disc diffusion method was used to determine the susceptibility of isolated bacteria to various antibiotics (oxfloxacin, pefloxacin, ciprorex, augumentin, gentamycin, ciproflox, septrin, ampicillin), while agar well diffusion method was used to determine the susceptibility of isolated fungi to some antifungal drugs (metronidazole, ketoconazole, itraconazole fluconazole). The antibacterial activity of the antibiotics used showed that ciproflux has the best inhibitory effect on all the test bacteria. Ketoconazole showed the highest inhibitory effect on the fungal isolates, followed by itraconazole, while metronidazole and fluconazole showed the least inhibitory effect on the entire test fungal isolates. Hence, the multiple drug resistance of most isolates to appropriate drugs of choice are of great public health concern and cells for periodic monitoring of antibiograms to detect possible changing patterns. Microbes isolated in the salt mining site can also be used as a source of gene(s) that can increase salt tolerance in different crop species through genetic engineering.

Keywords: microorganisms, antibacterial, antifungal, resistance, salt mining site, Ebonyi State

Procedia PDF Downloads 304
306 Biocontrol Potential of Trichoderma longibrachiatum as an Entomopathogenic Fungi against Bemisia tabaci

Authors: Waheed Anwar, Kiran Nawaz, Muhammad Saleem Haider, Ahmad Ali Shahid, Sehrish Iftikhar

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The whitefly, Bemisia tabaci (Gennadius), is a complex insect species, including many cryptic species or biotypes. Whitefly causes damage to many ornamental and horticultural crops through directly feeding on phloem sap, resulting in sooty mould and critically decreases the rate of photosynthesis of many host plants. Biological control has emerged as one of the most important methods for the management of soil-borne plant pathogens. Among the natural enemies of insects different entomopathogenic fungi are mostly used as biological control of the pest. The purpose of this research was to find indigenous insect-associated fungi and their virulence against Bemisia tabaci. A detailed survey of cotton fields in sample collection was conducted during July and August 2013 from the central mixed zone of Punjab, Pakistan. For the isolation of T. longibrachiatum, sabouraud dextrose peptone yeast extract agar (SDAY) media was used and morphological characterization of isolated T. longibrachiatum was studied using different dichotomous keys. Molecular Identification of the pathogen was confirmed by amplifying the internal transcribed spacer region. Blastn analysis showed 100% homology with already reported sequences on the database. For these bioassays, two conidial concentrations 4 × 108/mL & 4 × 104/mL of T. longibrachiatum was sprayed in clip cages for nymph and adult B. tabaci respectively under controlled environmental conditions. The pathogenicity of T. longibrachiatum was tested on nymph and adult whitefly to check mortality. Mortality of B. tabaci at nymphal and adult stages were observed after 24-hour intervals. Percentage mortality of nymphs treated with 4 x 104/mL conidia of T. longibrachiatum was 20, 24, 36 and 40% after 48, 72, 96, 72, 96, 120 and 144 hours respectively. However, no considerable difference was recorded in percentage mortality of whitefly after 120 and 144 hours. There were great variations after 24, 48, 72 and 96 hours in the rate of mortality. The efficacy of T. longibrachiatum as entomopathogenic fungi was evaluated in adult and nymphal stages of whitefly. Trichoderma longibrachiatum showed maximum activity on nymphal stages of whitefly as compared to adult stages. The percentage of conidial germination was also recorded on the outer surface of adult and nymphal stages of B. tabaci. The present findings indicated that T. longibrachiatum is an entomopathogenic fungus against B. tabaci and many species of Trichoderma were already reported as an antagonistc organism against a wide range of bacterial and fungal pathogens.

Keywords: efficacy, Trichoderma, virulence, bioassay

Procedia PDF Downloads 274
305 Investigations into the in situ Enterococcus faecalis Biofilm Removal Efficacies of Passive and Active Sodium Hypochlorite Irrigant Delivered into Lateral Canal of a Simulated Root Canal Model

Authors: Saifalarab A. Mohmmed, Morgana E. Vianna, Jonathan C. Knowles

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The issue of apical periodontitis has received considerable critical attention. Bacteria is integrated into communities, attached to surfaces and consequently form biofilm. The biofilm structure provides bacteria with a series protection skills against, antimicrobial agents and enhances pathogenicity (e.g. apical periodontitis). Sodium hypochlorite (NaOCl) has become the irrigant of choice for elimination of bacteria from the root canal system based on its antimicrobial findings. The aim of the study was to investigate the effect of different agitation techniques on the efficacy of 2.5% NaOCl to eliminate the biofilm from the surface of the lateral canal using the residual biofilm, and removal rate of biofilm as outcome measures. The effect of canal complexity (lateral canal) on the efficacy of the irrigation procedure was also assessed. Forty root canal models (n = 10 per group) were manufactured using 3D printing and resin materials. Each model consisted of two halves of an 18 mm length root canal with apical size 30 and taper 0.06, and a lateral canal of 3 mm length, 0.3 mm diameter located at 3 mm from the apical terminus. E. faecalis biofilms were grown on the apical 3 mm and lateral canal of the models for 10 days in Brain Heart Infusion broth. Biofilms were stained using crystal violet for visualisation. The model halves were reassembled, attached to an apparatus and tested under a fluorescence microscope. Syringe and needle irrigation protocol was performed using 9 mL of 2.5% NaOCl irrigant for 60 seconds. The irrigant was either left stagnant in the canal or activated for 30 seconds using manual (gutta-percha), sonic and ultrasonic methods. Images were then captured every second using an external camera. The percentages of residual biofilm were measured using image analysis software. The data were analysed using generalised linear mixed models. The greatest removal was associated with the ultrasonic group (66.76%) followed by sonic (45.49%), manual (43.97%), and passive irrigation group (control) (38.67%) respectively. No marked reduction in the efficiency of NaOCl to remove biofilm was found between the simple and complex anatomy models (p = 0.098). The removal efficacy of NaOCl on the biofilm was limited to the 1 mm level of the lateral canal. The agitation of NaOCl results in better penetration of the irrigant into the lateral canals. Ultrasonic agitation of NaOCl improved the removal of bacterial biofilm.

Keywords: 3D printing, biofilm, root canal irrigation, sodium hypochlorite

Procedia PDF Downloads 218
304 Zeolite 4A-confined Ni-Co Nanocluster: An Efficient and Durable Electrocatalyst for Alkaline Methanol Oxidation Reaction

Authors: Sarmistha Baruah, Akshai Kumar, Nageswara Rao Peela

Abstract:

The global energy crisis due to the dependence on fossil fuels and its limited reserves as well as environmental pollution are key concerns to the research communities. However, the implementation of alcohol-based fuel cells such as methanol is anticipated as a reliable source of future energy technology due to their high energy density, environment friendliness, ease of storage, transportation, etc. To drive the anodic methanol oxidation reaction (MOR) in direct methanol fuel cells (DMFCs), an active and long-lasting catalyst is necessary for efficient energy conversion from methanol. Recently, transition metal-zeolite-based materials have been considered versatile catalysts for a variety of industrial and lab-scale processes. Large specific surface area, well-organized micropores, and adjustable acidity/basicity are characteristics of zeolites that make them excellent supports for immobilizing small-sized and highly dispersed metal species. Significant advancement in the production and characterization of well-defined metal clusters encapsulated within zeolite matrix has substantially expanded the library of materials available, and consequently, their catalytic efficacy. In this context, we developed bimetallic Ni-Co catalysts encapsulated within LTA (also known as 4A) zeolite via a method combined with the in-situ encapsulation of metal species using hydrothermal treatment followed by a chemical reduction process. The prepared catalyst was characterized using advanced characterization techniques, such as X-ray diffraction (XRD), field emission transmission electron microscope (FETEM), field emission scanning electron microscope (FESEM), energy dispersive X-ray (EDX), and X-ray photoelectron spectroscopy (XPS). The electrocatalytic activity of the catalyst for MOR was carried out in an alkaline medium at room temperature using techniques such as cyclic voltammetry (CV), and chronoamperometry (CA). The resulting catalyst exhibited better catalytic activity of 12.1 mA cm-2 at 1.12 V vs Ag/AgCl and retained remarkable stability (~77%) even after 1000 cycles CV test for the electro-oxidation of methanol in alkaline media without any significant microstructural changes. The high surface area, better Ni-Co species integration in the zeolite, and the ample amount of surface hydroxyl groups contribute to highly dispersed active sites and quick analyte diffusion, which provide notable MOR kinetics. Thus, this study will open up new possibilities to develop a noble metal-free zeolite-based electrocatalyst due to its simple synthesis steps, large-scale fabrication, improved stability, and efficient activity for DMFC application.

Keywords: alkaline media, bimetallic, encapsulation, methanol oxidation reaction, LTA zeolite.

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303 Degradation and Detoxification of Tetracycline by Sono-Fenton and Ozonation

Authors: Chikang Wang, Jhongjheng Jian, Poming Huang

Abstract:

Among a wide variety of pharmaceutical compounds, tetracycline antibiotics are one of the largest groups of pharmaceutical compounds extensively used in human and veterinary medicine to treat and prevent bacterial infections. Because it is water soluble, biologically active, stable and bio-refractory, release to the environment threatens aquatic life and increases the risk posed by antibiotic-resistant pathogens. In practice, due to its antibacterial nature, tetracycline cannot be effectively destructed by traditional biological methods. Hence, in this study, two advanced oxidation processes such as ozonation and sono-Fenton processes were conducted individually to degrade the tetracycline for investigating their feasibility on tetracycline degradation. Effect of operational variables on tetracycline degradation, release of nitrogen and change of toxicity were also proposed. Initial tetracycline concentration was 50 mg/L. To evaluate the efficiency of tetracycline degradation by ozonation, the ozone gas was produced by an ozone generator (Model LAB2B, Ozonia) and introduced into the reactor with different flows (25 - 500 mL/min) at varying pH levels (pH 3 - pH 11) and reaction temperatures (15 - 55°C). In sono-Fenton system, an ultrasonic transducer (Microson VCX 750, USA) operated at 20 kHz combined with H₂O₂ (2 mM) and Fe²⁺ (0.2 mM) were carried out at different pH levels (pH 3 - pH 11), aeration gas and flows (air and oxygen; 0.2 - 1.0 L/min), tetracycline concentrations (10 - 200 mg/L), reaction temperatures (15 - 55°C) and ultrasonic powers (25 - 200 Watts), respectively. Sole ultrasound was ineffective on tetracycline degradation, where the degradation efficiencies were lower than 10% with 60 min reaction. Contribution of Fe²⁺ and H₂O₂ on the degradation of tetracycline was significant, where the maximum tetracycline degradation efficiency in sono-Fenton process was as high as 91.3% followed by 45.8% mineralization. Effect of initial pH level on tetracycline degradation was insignificant from pH 3 to pH 6 but significantly decreased as the pH was greater than pH 7. Increase of the ultrasonic power was slightly increased the degradation efficiency of tetracycline, which indicated that the hydroxyl radicals dominated the oxidation of tetracycline. Effects of aeration of air or oxygen with different flows and reaction temperatures were insignificant. Ozonation showed better efficiencies in tetracycline degradation, where the optimum reaction condition was found at pH 3, 100 mL O₃/min and 25°C with 94% degradation and 60% mineralization. The toxicity of tetracycline was significantly decreased due to the mineralization of tetracycline. In addition, less than 10% of nitrogen content was released to solution phase as NH₃-N, and the most degraded tetracycline cannot be full mineralized to CO₂. The results shown in this study indicated that both the sono-Fenton process and ozonation can effectively degrade the tetracycline and reduce its toxicity at profitable condition. The costs of two systems needed to be further investigated to understand the feasibility in tetracycline degradation.

Keywords: degradation, detoxification, mineralization, ozonation, sono-Fenton process, tetracycline

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302 2,7-Diazaindole as a Photophysical Probe for Excited State Hydrogen/Proton Transfer

Authors: Simran Baweja, Bhavika Kalal, Surajit Maity

Abstract:

Photoinduced tautomerization reactions have been the centre of attention among the scientific community over the past several decades because of their significance in various biological systems. 7-azaindole (7AI) is considered a model system for DNA base pairing and to understand the role of such tautomerization reactions in mutations. To the best of our knowledge, extensive studies have been carried out on 7-azaindole and its solvent clusters exhibiting proton/ hydrogen transfer in both solution as well as gas phases. Derivatives of the above molecule, like 2,7- and 2,6-diazaindoles are proposed to have even better photophysical properties due to the presence of -aza group on the 2nd position. However, there are studies in the solution phase that suggest the relevance of these molecules, but there are no experimental studies reported in the gas phase yet. In our current investigation, we present the first gas phase spectroscopic data of 2,7-diazaindole (2,7-DAI) and its solvent cluster (2,7-DAI-H2O). In this, we have employed state-of-the-art laser spectroscopic methods such as fluorescence excitation (LIF), dispersed fluorescence (DF), resonant two-photon ionization-time of flight mass spectrometry (2C-R2PI), photoionization efficiency spectroscopy (PIE), IR-UV double resonance spectroscopy, i.e., fluorescence-dip infrared spectroscopy (FDIR) and resonant ion-dip infrared spectroscopy (IDIR) to understand the electronic structure of the molecule. The origin band corresponding to the S1 ← S0 transition of the bare 2,7-DAI is found to be positioned at 33910 cm-1, whereas the origin band corresponding to S1 ← S0 transition of the 2,7-DAI-H2O is positioned at 33074 cm-1. The red-shifted transition in the case of solvent cluster suggests the enhanced feasibility of excited state hydrogen/ proton transfer. The ionization potential for the 2,7-DAI molecule is found to be 8.92 eV which is significantly higher than the previously reported 7AI (8.11 eV) molecule, making it a comparatively complex molecule to study. The ionization potential is reduced by 0.14 eV in the case of 2,7-DAI-H2O (8.78 eV) cluster compared to that of 2,7-DAI. Moreover, on comparison with the available literature values of 7AI, we found the origin band of 2,7-DAI and 2,7-DAI-H2O to be red-shifted by -729 and -280 cm-1 respectively. The ground and excited state N-H stretching frequencies of the 27DAI molecule were determined using fluorescence-dip infrared spectra (FDIR) and resonant ion dip infrared spectroscopy (IDIR), obtained at 3523 and 3467 cm-1, respectively. The lower value of vNH in the electronically excited state of 27DAI implies the higher acidity of the group compared to the ground state. Moreover, we have done extensive computational analysis, which suggests that the energy barrier in the excited state reduces significantly as we increase the number of catalytic solvent molecules (S= H2O, NH3) as well as the polarity of solvent molecules. We found that the ammonia molecule is a better candidate for hydrogen transfer compared to water because of its higher gas-phase basicity. Further studies are underway to understand the excited state dynamics and photochemistry of such N-rich chromophores.

Keywords: excited state hydrogen transfer, supersonic expansion, gas phase spectroscopy, IR-UV double resonance spectroscopy, laser induced fluorescence, photoionization efficiency spectroscopy

Procedia PDF Downloads 64