Search results for: cell morphology prediction
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 7028

Search results for: cell morphology prediction

4208 Cost-Conscious Treatment of Basal Cell Carcinoma

Authors: Palak V. Patel, Jessica Pixley, Steven R. Feldman

Abstract:

Introduction: Basal cell carcinoma (BCC) is the most common skin cancer worldwide and requires substantial resources to treat. When choosing between indicated therapies, providers consider their associated adverse effects, efficacy, cosmesis, and function preservation. The patient’s tumor burden, infiltrative risk, and risk of tumor recurrence are also considered. Treatment cost is often left out of these discussions. This can lead to financial toxicity, which describes the harm and quality of life reductions inflicted by high care costs. Methods: We studied the guidelines set forth by the American Academy of Dermatology for the treatment of BCC. A PubMed literature search was conducted to identify the costs of each recommended therapy. We discuss costs alongside treatment efficacy and side-effect profile. Results: Surgical treatment for BCC can be cost-effective if the appropriate treatment is selected for the presenting tumor. Curettage and electrodesiccation can be used in low-grade, low-recurrence tumors in aesthetically unimportant areas. The benefits of cost-conscious care are not likely to be outweighed by the risks of poor cosmesis or tumor return ($471 BCC of the cheek). When tumor burden is limited, MMS offers better cure rates and lower recurrence rates than surgical excision, and with comparable costs (MMS $1263; SE $949). Surgical excision with permanent sections may be indicated when tumor burden is more extensive or if molecular testing is necessary. The utility of surgical excision with frozen sections, which costs substantially more than MMS without comparable outcomes, is less clear (SE with frozen sections $2334-$3085). Less data exists on non-surgical treatments for BCC. These techniques cost less, but recurrence-risk is high. Side-effects of nonsurgical treatment are limited to local skin reactions, and cosmesis is good. Cryotherapy, 5-FU, and MAL-PDT are all more affordable than surgery, but high recurrence rates increase risk of secondary financial and psychosocial burden (recurrence rates 21-39%; cost $100-270). Radiation therapy offers better clearance rates than other nonsurgical treatments but is associated with similar recurrence rates and a significantly larger financial burden ($2591-$3460 BCC of the cheek). Treatments for advanced or metastatic BCC are extremely costly, but few patients require their use, and the societal cost burden remains low. Vismodegib and sonidegib have good response rates but substantial side effects, and therapy should be combined with multidisciplinary care and palliative measures. Expert-review has found sonidegib to be the less expensive and more efficacious option (vismodegib $128,358; sonidegib $122,579). Platinum therapy, while not FDA-approved, is also effective but expensive (~91,435). Immunotherapy offers a new line of treatment in patients intolerant of hedgehog inhibitors ($683,061). Conclusion: Dermatologists working within resource-compressed practices and with resource-limited patients must prudently manage the healthcare dollar. Surgical therapies for BCC offer the lowest risk of recurrence at the most reasonable cost. Non-surgical therapies are more affordable, but high recurrence rates increase the risk of secondary financial and psychosocial burdens. Treatments for advanced BCC are incredibly costly, but the low incidence means the overall cost to the system is low.

Keywords: nonmelanoma skin cancer, basal cell skin cancer, squamous cell skin cancer, cost of care

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4207 Influence of Intra-Yarn Permeability on Mesoscale Permeability of Plain Weave and 3D Fabrics

Authors: Debabrata Adhikari, Mikhail Matveev, Louise Brown, Andy Long, Jan Kočí

Abstract:

A good understanding of mesoscale permeability of complex architectures in fibrous porous preforms is of particular interest in order to achieve efficient and cost-effective resin impregnation of liquid composite molding (LCM). Fabrics used in structural reinforcements are typically woven or stitched. However, 3D fabric reinforcement is of particular interest because of the versatility in the weaving pattern with the binder yarn and in-plain yarn arrangements to manufacture thick composite parts, overcome the limitation in delamination, improve toughness etc. To predict the permeability based on the available pore spaces between the inter yarn spaces, unit cell-based computational fluid dynamics models have been using the Stokes Darcy model. Typically, the preform consists of an arrangement of yarns with spacing in the order of mm, wherein each yarn consists of thousands of filaments with spacing in the order of μm. The fluid flow during infusion exchanges the mass between the intra and inter yarn channels, meaning there is no dead-end of flow between the mesopore in the inter yarn space and the micropore in the yarn. Several studies have employed the Brinkman equation to take into account the flow through dual-scale porosity reinforcement to estimate their permeability. Furthermore, to reduce the computational effort of dual scale flow, scale separation criteria based on the ratio between yarn permeability to the yarn spacing was also proposed to quantify the dual scale and negligible micro-scale flow regime for the prediction of mesoscale permeability. In the present work, the key parameter to identify the influence of intra yarn permeability on the mesoscale permeability has been investigated with the systematic study of weft and warp yarn spacing on the plane weave as well as the position of binder yarn and number of in-plane yarn layers on 3D weave fabric. The permeability tensor has been estimated using an OpenFOAM-based model for the various weave pattern with idealized geometry of yarn implemented using open-source software TexGen. Additionally, scale separation criterion has been established based on the various configuration of yarn permeability for the 3D fabric with both the isotropic and anisotropic yarn from Gebart’s model. It was observed that the variation of mesoscale permeability Kxx within 30% when the isotropic porous yarn is considered for a 3D fabric with binder yarn. Furthermore, the permeability model developed in this study will be used for multi-objective optimizations of the preform mesoscale geometry in terms of yarn spacing, binder pattern, and a number of layers with an aim to obtain improved permeability and reduced void content during the LCM process.

Keywords: permeability, 3D fabric, dual-scale flow, liquid composite molding

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4206 Stent Surface Functionalisation via Plasma Treatment to Promote Fast Endothelialisation

Authors: Irene Carmagnola, Valeria Chiono, Sandra Pacharra, Jochen Salber, Sean McMahon, Chris Lovell, Pooja Basnett, Barbara Lukasiewicz, Ipsita Roy, Xiang Zhang, Gianluca Ciardelli

Abstract:

Thrombosis and restenosis after stenting procedure can be prevented by promoting fast stent wall endothelialisation. It is well known that surface functionalisation with antifouling molecules combining with extracellular matrix proteins is a promising strategy to design biomimetic surfaces able to promote fast endothelialization. In particular, REDV has gained much attention for the ability to enhance rapid endothelialization due to its specific affinity with endothelial cells (ECs). In this work, a two-step plasma treatment was performed to polymerize a thin layer of acrylic acid, used to subsequently graft PEGylated-REDV and polyethylene glycol (PEG) at different molar ratio with the aim to selectively promote endothelial cell adhesion avoiding platelet activation. PEGylate-REDV was provided by Biomatik and it is formed by 6 PEG monomer repetitions (Chempep Inc.), with an NH2 terminal group. PEG polymers were purchased from Chempep Inc. with two different chain lengths: m-PEG6-NH2 (295.4 Da) with 6 monomer repetitions and m-PEG12-NH2 (559.7 Da) with 12 monomer repetitions. Plasma activation was obtained by operating at 50W power, 5 min of treatment and at an Ar flow rate of 20 sccm. Pure acrylic acid (99%, AAc) vapors were diluted in Ar (flow = 20 sccm) and polymerized by a pulsed plasma discharge applying a discharge RF power of 200 W, a duty cycle of 10% (on time = 10 ms, off time = 90 ms) for 10 min. After plasma treatment, samples were dipped into an 1-(3-dimethylaminopropyl)-3- ethylcarbodiimide (EDC)/N-hydroxysuccinimide (NHS) solution (ratio 4:1, pH 5.5) for 1 h at 4°C and subsequently dipped in PEGylate-REDV and PEGylate-REDV:PEG solutions at different molar ratio (100 μg/mL in PBS) for 20 h at room temperature. Surface modification was characterized through physico-chemical analyses and in vitro cell tests. PEGylated-REDV peptide and PEG were successfully bound to the carboxylic groups that are formed on the polymer surface after plasma reaction. FTIR-ATR spectroscopy, X -ray Photoelectron Spectroscopy (XPS) and contact angle measurement gave a clear indication of the presence of the grafted molecules. The use of PEG as a spacer allowed for an increase in wettability of the surface, and the effect was more evident by increasing the amount of PEG. Endothelial cells adhered and spread well on the surfaces functionalized with the REDV sequence. In conclusion, a selective coating able to promote a new endothelial cell layer on polymeric stent surface was developed. In particular, a thin AAc film was polymerised on the polymeric surface in order to expose –COOH groups, and PEGylate-REDV and PEG were successful grafted on the polymeric substrates. The REDV peptide demonstrated to encourage cell adhesion with a consequent, expected improvement of the hemocompatibility of these polymeric surfaces in vivo. Acknowledgements— This work was funded by the European Commission 7th Framework Programme under grant agreement number 604251- ReBioStent (Reinforced Bioresorbable Biomaterials for Therapeutic Drug Eluting Stents). The authors thank all the ReBioStent partners for their support in this work.

Keywords: endothelialisation, plasma treatment, stent, surface functionalisation

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4205 Performance and Limitations of Likelihood Based Information Criteria and Leave-One-Out Cross-Validation Approximation Methods

Authors: M. A. C. S. Sampath Fernando, James M. Curran, Renate Meyer

Abstract:

Model assessment, in the Bayesian context, involves evaluation of the goodness-of-fit and the comparison of several alternative candidate models for predictive accuracy and improvements. In posterior predictive checks, the data simulated under the fitted model is compared with the actual data. Predictive model accuracy is estimated using information criteria such as the Akaike information criterion (AIC), the Bayesian information criterion (BIC), the Deviance information criterion (DIC), and the Watanabe-Akaike information criterion (WAIC). The goal of an information criterion is to obtain an unbiased measure of out-of-sample prediction error. Since posterior checks use the data twice; once for model estimation and once for testing, a bias correction which penalises the model complexity is incorporated in these criteria. Cross-validation (CV) is another method used for examining out-of-sample prediction accuracy. Leave-one-out cross-validation (LOO-CV) is the most computationally expensive variant among the other CV methods, as it fits as many models as the number of observations. Importance sampling (IS), truncated importance sampling (TIS) and Pareto-smoothed importance sampling (PSIS) are generally used as approximations to the exact LOO-CV and utilise the existing MCMC results avoiding expensive computational issues. The reciprocals of the predictive densities calculated over posterior draws for each observation are treated as the raw importance weights. These are in turn used to calculate the approximate LOO-CV of the observation as a weighted average of posterior densities. In IS-LOO, the raw weights are directly used. In contrast, the larger weights are replaced by their modified truncated weights in calculating TIS-LOO and PSIS-LOO. Although, information criteria and LOO-CV are unable to reflect the goodness-of-fit in absolute sense, the differences can be used to measure the relative performance of the models of interest. However, the use of these measures is only valid under specific circumstances. This study has developed 11 models using normal, log-normal, gamma, and student’s t distributions to improve the PCR stutter prediction with forensic data. These models are comprised of four with profile-wide variances, four with locus specific variances, and three which are two-component mixture models. The mean stutter ratio in each model is modeled as a locus specific simple linear regression against a feature of the alleles under study known as the longest uninterrupted sequence (LUS). The use of AIC, BIC, DIC, and WAIC in model comparison has some practical limitations. Even though, IS-LOO, TIS-LOO, and PSIS-LOO are considered to be approximations of the exact LOO-CV, the study observed some drastic deviations in the results. However, there are some interesting relationships among the logarithms of pointwise predictive densities (lppd) calculated under WAIC and the LOO approximation methods. The estimated overall lppd is a relative measure that reflects the overall goodness-of-fit of the model. Parallel log-likelihood profiles for the models conditional on equal posterior variances in lppds were observed. This study illustrates the limitations of the information criteria in practical model comparison problems. In addition, the relationships among LOO-CV approximation methods and WAIC with their limitations are discussed. Finally, useful recommendations that may help in practical model comparisons with these methods are provided.

Keywords: cross-validation, importance sampling, information criteria, predictive accuracy

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4204 Phenotype Prediction of DNA Sequence Data: A Machine and Statistical Learning Approach

Authors: Mpho Mokoatle, Darlington Mapiye, James Mashiyane, Stephanie Muller, Gciniwe Dlamini

Abstract:

Great advances in high-throughput sequencing technologies have resulted in availability of huge amounts of sequencing data in public and private repositories, enabling a holistic understanding of complex biological phenomena. Sequence data are used for a wide range of applications such as gene annotations, expression studies, personalized treatment and precision medicine. However, this rapid growth in sequence data poses a great challenge which calls for novel data processing and analytic methods, as well as huge computing resources. In this work, a machine and statistical learning approach for DNA sequence classification based on $k$-mer representation of sequence data is proposed. The approach is tested using whole genome sequences of Mycobacterium tuberculosis (MTB) isolates to (i) reduce the size of genomic sequence data, (ii) identify an optimum size of k-mers and utilize it to build classification models, (iii) predict the phenotype from whole genome sequence data of a given bacterial isolate, and (iv) demonstrate computing challenges associated with the analysis of whole genome sequence data in producing interpretable and explainable insights. The classification models were trained on 104 whole genome sequences of MTB isoloates. Cluster analysis showed that k-mers maybe used to discriminate phenotypes and the discrimination becomes more concise as the size of k-mers increase. The best performing classification model had a k-mer size of 10 (longest k-mer) an accuracy, recall, precision, specificity, and Matthews Correlation coeffient of 72.0%, 80.5%, 80.5%, 63.6%, and 0.4 respectively. This study provides a comprehensive approach for resampling whole genome sequencing data, objectively selecting a k-mer size, and performing classification for phenotype prediction. The analysis also highlights the importance of increasing the k-mer size to produce more biological explainable results, which brings to the fore the interplay that exists amongst accuracy, computing resources and explainability of classification results. However, the analysis provides a new way to elucidate genetic information from genomic data, and identify phenotype relationships which are important especially in explaining complex biological mechanisms.

Keywords: AWD-LSTM, bootstrapping, k-mers, next generation sequencing

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4203 Phenotype Prediction of DNA Sequence Data: A Machine and Statistical Learning Approach

Authors: Darlington Mapiye, Mpho Mokoatle, James Mashiyane, Stephanie Muller, Gciniwe Dlamini

Abstract:

Great advances in high-throughput sequencing technologies have resulted in availability of huge amounts of sequencing data in public and private repositories, enabling a holistic understanding of complex biological phenomena. Sequence data are used for a wide range of applications such as gene annotations, expression studies, personalized treatment and precision medicine. However, this rapid growth in sequence data poses a great challenge which calls for novel data processing and analytic methods, as well as huge computing resources. In this work, a machine and statistical learning approach for DNA sequence classification based on k-mer representation of sequence data is proposed. The approach is tested using whole genome sequences of Mycobacterium tuberculosis (MTB) isolates to (i) reduce the size of genomic sequence data, (ii) identify an optimum size of k-mers and utilize it to build classification models, (iii) predict the phenotype from whole genome sequence data of a given bacterial isolate, and (iv) demonstrate computing challenges associated with the analysis of whole genome sequence data in producing interpretable and explainable insights. The classification models were trained on 104 whole genome sequences of MTB isoloates. Cluster analysis showed that k-mers maybe used to discriminate phenotypes and the discrimination becomes more concise as the size of k-mers increase. The best performing classification model had a k-mer size of 10 (longest k-mer) an accuracy, recall, precision, specificity, and Matthews Correlation coeffient of 72.0 %, 80.5 %, 80.5 %, 63.6 %, and 0.4 respectively. This study provides a comprehensive approach for resampling whole genome sequencing data, objectively selecting a k-mer size, and performing classification for phenotype prediction. The analysis also highlights the importance of increasing the k-mer size to produce more biological explainable results, which brings to the fore the interplay that exists amongst accuracy, computing resources and explainability of classification results. However, the analysis provides a new way to elucidate genetic information from genomic data, and identify phenotype relationships which are important especially in explaining complex biological mechanisms

Keywords: AWD-LSTM, bootstrapping, k-mers, next generation sequencing

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4202 Chrysin-Loaded PLGA-PEG Nanoparticles Designed for Enhanced Inhibitory Effect on the Breast Cancer Cell Line

Authors: Faraz Zarghami, Elham Anari, Nosratollah Zarghami, Yones Pilehvar-Soltanahmadi, Abolfazl Akbarzadeh, Sepideh Jalilzadeh-Tabrizi

Abstract:

The development of nanotherapy has presented a new method of drug delivery targeted directly to the neoplasmic tissues, to maximize the action with fewer dose requirements. In the past two decades, poly(lactic-co-glycolic acid) (PLGA) has frequently been investigated by many researchers and is a popular polymeric candidate, due to its biocompatibility and biodegradability, exhibition of a wide range of erosion times, tunable mechanical properties, and most notably, because it is a FDA-approved polymer. Chrysin is a natural flavonoid which has been reported to have some significant biological effects on the processes of chemical defense, nitrogen fixation, inflammation, and oxidation. However, the low solubility in water decreases its bioavailability and consequently disrupts the biomedical benefits. Being loaded with PLGA-PEG increases chrysin solubility and drug tolerance, and decreases the discordant effects of the drug. The well-structured chrysin efficiently accumulates in the breast cancer cell line (T47D). In the present study, the structure and chrysin loading were delineated using proton nuclear magnetic resonance (HNMR), Fourier-transform infrared spectroscopy (FT-IR), and scanning electron microscopy (SEM), and the in vitro cytotoxicity of pure and nanochrysin was studied by the MTT assay. Next, the RNA was exploited and the cytotoxic effects of chrysin were studied by real-time PCR. In conclusion, the nanochrysin therapy developed is a novel method that could increase cytotoxicity to cancer cells without damaging the normal cells, and would be promising in breast cancer therapy.

Keywords: MTT assay, chrysin, flavonoids, nanotherapy

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4201 Pharmacognostical and Phytochemical Investigation of the Endemic Medicinal Plant Tekchebilium arvensis Linn

Authors: K. Bengango, H. Mesahsah, F. Haseb-Reho, J. M. Tafrate

Abstract:

This present work was conducted to explore the micro-morphology and phytochemical characterization of the endemic medicinal plant Tekchebilium arvensis Linn (Asteraceae). Macroscopy, microscopy, physicochemical analysis and WHO recommended parameters for standardizations were performed. Microscopic evaluation revealed the presence of abaxial epidermis with paracytic stomata. Petiole showed epidermis, vascular strands, ground tissue and secretary cavities. Physico-chemical tests like ash values, loss on drying, extractive values were determined. Preliminary phytochemical screening showed the presence of sterols, tannins, flavonoids, glycosides, volatile oil, terpenoids, saponin and alkaloids.

Keywords: Tekchebilium arvensis Linn, Asteraceae, microscopical evaluation, phytochemical, powder microscopy, standardization

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4200 Prediction of Formation Pressure Using Artificial Intelligence Techniques

Authors: Abdulmalek Ahmed

Abstract:

Formation pressure is the main function that affects drilling operation economically and efficiently. Knowing the pore pressure and the parameters that affect it will help to reduce the cost of drilling process. Many empirical models reported in the literature were used to calculate the formation pressure based on different parameters. Some of these models used only drilling parameters to estimate pore pressure. Other models predicted the formation pressure based on log data. All of these models required different trends such as normal or abnormal to predict the pore pressure. Few researchers applied artificial intelligence (AI) techniques to predict the formation pressure by only one method or a maximum of two methods of AI. The objective of this research is to predict the pore pressure based on both drilling parameters and log data namely; weight on bit, rotary speed, rate of penetration, mud weight, bulk density, porosity and delta sonic time. A real field data is used to predict the formation pressure using five different artificial intelligence (AI) methods such as; artificial neural networks (ANN), radial basis function (RBF), fuzzy logic (FL), support vector machine (SVM) and functional networks (FN). All AI tools were compared with different empirical models. AI methods estimated the formation pressure by a high accuracy (high correlation coefficient and low average absolute percentage error) and outperformed all previous. The advantage of the new technique is its simplicity, which represented from its estimation of pore pressure without the need of different trends as compared to other models which require a two different trend (normal or abnormal pressure). Moreover, by comparing the AI tools with each other, the results indicate that SVM has the advantage of pore pressure prediction by its fast processing speed and high performance (a high correlation coefficient of 0.997 and a low average absolute percentage error of 0.14%). In the end, a new empirical correlation for formation pressure was developed using ANN method that can estimate pore pressure with a high precision (correlation coefficient of 0.998 and average absolute percentage error of 0.17%).

Keywords: Artificial Intelligence (AI), Formation pressure, Artificial Neural Networks (ANN), Fuzzy Logic (FL), Support Vector Machine (SVM), Functional Networks (FN), Radial Basis Function (RBF)

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4199 Chitosan Doped Curcumin Gold Clusters Flexible Nanofiber for Wound Dressing and Anticancer Activities

Authors: Saravanan Govindaraju, Kyusik Yun

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The purpose of this study is to develop the chitosan doped curcumin gold cluster nanofiber for wound healing and skin cancer drug delivery applications. Chitosan is a typical marine polysaccharide composed of glucosamine and n-acetyl glucosamine biodegradable and biocompatible polymer. Curcumin is a natural bioactive molecule obtained from Curcuma longo, it mostly occurs in some Asian countries like India and China. It has naturally antioxidant, antimicrobial, wound healing and anticancer property. Due to this advantage, we prepared a combination of natural polymer chitosan with Curcumin and gold nanocluster nanofiber (CH-CUR-AuNCs nanofibers). The prepared nanofiber was characterized by using Fourier transform infrared spectroscopy (FT-IR), and scanning electron microscopy (SEM). Antibacterial studies were performed with E.coli and S.aureus. Antioxidant assay, drug release test, and cytotoxicity will be evaluated. Prepared nanofiber emits low intensity of red fluorescent. The FTIR confirm the presence of chitosan and Curcumin in the nanofiber. In vitro study clearly shows the antibacterial activity against the gram negative and gram positive bacteria. Particularly, synthesised nanofibers provide better antibacterial activity against gram negative than gram positive. Cytotoxicity study also provides better killing rate in cancer cell, biocompatible with normal cell. Prepared CH-CUR-AuNCs nanofibers provide the better killing rate to bacterial strains and cancer cells. Finally, prepared nanofiber can be possible to use for wound healing dressing, patch for skin cancer and other biomedical applications.

Keywords: curcumin, chitosan, gold clusters, nanofibers

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4198 Analysis of the Detachment of Water Droplets from a Porous Fibrous Surface

Authors: Ibrahim Rassoul, E-K. Si Ahmed

Abstract:

The growth, deformation, and detachment of fluid droplets adherent to solid substrates is a problem of fundamental interest with numerous practical applications. Specific interest in this proposal is the problem of a droplet on a fibrous, hydrophobic substrate subjected to body or external forces (gravity, convection). The past decade has seen tremendous advances in proton exchange membrane fuel cell (PEMFC) technology. However, there remain many challenges to bring commercially viable stationary PEMFC products to the market. PEMFCs are increasingly emerging as a viable alternative clean power source for automobile and stationary applications. Before PEMFCs can be employed to power automobiles and homes, several key technical challenges must be properly addressed. One technical challenge is elucidating the mechanisms underlying water transport in and removal from PEMFCs. On the one hand, sufficient water is needed in the polymer electrolyte membrane or PEM to maintain sufficiently high proton conductivity. On the other hand, too much liquid water present in the cathode can cause 'flooding' (that is, pore space is filled with excessive liquid water) and hinder the transport of the oxygen reactant from the gas flow channel (GFC) to the three-phase reaction sites. The aim of this work is to investigate the stability of a liquid water droplet emerging form a GDL pore, to gain fundamental insight into the instability process leading to detachment. The approach will combine analytical and numerical modeling with experimental visualization and measurements.

Keywords: polymer electrolyte fuel cell, water droplet, gas diffusion layer, contact angle, surface tension

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4197 Efficient Photodegradation of Methyl Red Dye by Kaolin Clay Supported Zinc Oxide Nanoparticles with Their Antibacterial and Antioxidant Activities

Authors: Idrees Khan, Zhang Baoliang

Abstract:

Kaolin clay (KC) supported Zinc oxide (ZnO/KC) and ZnO nanoparticles (NPs) were prepared by a chemical reduction process and used for the photodegradation of methyl red (MR) as photocatalysts. Due to the interlayered porous structure of KC, we achieved a perfect association between ZnO NPs and KC. SEM image showed the irregular morphology of ZnO NPs, while ZnO/KC NCs were predominately round-shaped. Moreover, in both cases, NPs were present in dispersed and agglomerated forms with an average particle size way below 100 nm. The results acquired from photodegradation analyses showed that ZnO NPs and ZnO/KC NCs degraded about 82% and 99% of MR under UV light in a short irradiation time within 10 min. The recovered and re-recovered ZnO NPs and ZnO/KC NCs were also considerably photodegraded MR in an aqueous medium. The same NPs also exhibit promising bioactivities against two pathogenic bacteria, i.e., Citrobacter and Providencia. ZnO/KC NCs' antioxidant activity reached a reasonable 70% compared to the 88% activity of the standard ascorbic acid.

Keywords: nanoparticles, photocatalyst, photodegradation, zinc oxide, methyl red

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4196 Hydroxyapatite Based Porous Scaffold for Tooth Tissue Engineering

Authors: Pakize Neslihan Taslı, Alev Cumbul, Gul Merve Yalcın, Fikrettin Sahin

Abstract:

A key experimental trial in the regeneration of large oral and craniofacial defects is the neogenesis of osseous and ligamentous interfacial structures. Currently, oral regenerative medicine strategies are unpredictable for repair of tooth supporting tissues destroyed as a consequence of trauma, chronic infection or surgical resection. A different approach combining the gel-casting method with Hydroxy Apatite HA-based scaffold and different cell lineages as a hybrid system leads to successively mimic the early stage of tooth development, in vitro. HA is widely accepted as a bioactive material for guided bone and tooth regeneration. In this study, it was reported that, HA porous scaffold preparation, characterization and evaluation of structural and chemical properties. HA is the main factor that exists in tooth and it is in harmony with structural, biological, and mechanical characteristics. Here, this study shows mimicking immature tooth at the late bell stage design and construction of HA scaffolds for cell transplantation of human Adipose Stem Cells (hASCs), human Bone Marrow Stem Cells (hBMSCs) and Gingival Epitelial cells for the formation of human tooth dentin-pulp-enamel complexes in vitro. Scaffold characterization was demonstrated by SEM, FTIR and pore size and density measurements. The biological contraction of dental tissues against each other was demonstrated by mRNA gene expressions, histopatologic observations and protein release profile by ELISA tecnique. The tooth shaped constructs with a pore size ranging from 150 to 300 µm arranged by gathering right amounts of materials provide interconnected macro-porous structure. The newly formed tissue like structures that grow and integrate within the HA designed constructs forming tooth cementum like tissue, pulp and bone structures. These findings are important as they emphasize the potential biological effect of the hybrid scaffold system. In conclusion, this in vitro study clearly demonstrates that designed 3D scaffolds shaped as a immature tooth at the late bell stage were essential to form enamel-dentin-pulp interfaces with an appropriate cell and biodegradable material combination. The biomimetic architecture achieved here is providing a promising platform for dental tissue engineering.

Keywords: tooth regeneration, tissue engineering, adipose stem cells, hydroxyapatite tooth engineering, porous scaffold

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4195 Right Atrial Tissue Morphology in Acquired Heart Diseases

Authors: Edite Kulmane, Mara Pilmane, Romans Lacis

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Introduction: Acquired heart diseases remain one of the leading health care problems in the world. Changes in myocardium of the diseased hearts are complex and pathogenesis is still not fully clear. The aim of this study was to identify appearance and distribution of apoptosis, homeostasis regulating factors, and innervation and ischemia markers in right atrial tissue in different acquired heart diseases. Methods: During elective open heart surgery were taken right atrial tissue fragments from 12 patients. All patients were operated because of acquired heart diseases- aortic valve stenosis (5 patients), coronary heart disease (5 patients), coronary heart disease and secondary mitral insufficiency (1 patient) and mitral disease (1 patient). The mean age was (mean±SD) 70,2±7,0 years (range 58-83 years). The tissues were stained with haematoxylin and eosin methods for routine light-microscopical examination and for immunohistochemical detection of protein gene peptide 9.5 (PGP 9.5), human atrial natriuretic peptide (hANUP), vascular endothelial growth factor (VEGF), chromogranin A and endothelin. Apoptosis was detected by TUNEL method. Results: All specimens showed degeneration of cardiomyocytes with lysis of myofibrils, diffuse vacuolization especially in perinuclear region, different size of cells and their nuclei. The severe invasion of connective tissue was observed in main part of all fragments. The apoptotic index ranged from 24 to 91%. One specimen showed region of newly performed microvessels with cube shaped endotheliocytes that were positive for PGP 9.5, endothelin, chromogranin A and VEGF. From all fragments, taken from patients with coronary heart disease, there were observed numerous PGP 9.5-containing nerve fibres, except in patient with secondary mitral insufficiency, who showed just few PGP 9.5 positive nerves. In majority of specimens there were regions observed with cube shaped mixed -VEGF immunoreactive endocardial and epicardial cells. Only VEGF positive endothelial cells were observed just in few specimens. There was no significant difference of hANUP secreting cells among all specimens. All patients operated due to the coronary heart disease moderate to numerous number of chromogranin A positive cells were seen while in patients with aortic valve stenosis tissue demonstrated just few factor positive cells. Conclusions: Complex detection of different factors may indicate selectively disordered morphopathogenetical event of heart disease: decrease of PGP 9.5 nerves suggests the decreased innervation of organ; increased apoptosis indicates the cell death without ingrowth of connective tissue; persistent presence of hANUP proves the unchanged homeostasis of cardiomyocytes probably supported by expression of chromogranins. Finally, decrease of VEGF detects the regions of affected blood vessels in heart affected by acquired heart disease.

Keywords: heart, apoptosis, protein-gene peptide 9.5, atrial natriuretic peptide, vascular endothelial growth factor, chromogranin A, endothelin

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4194 Flexible and Color Tunable Inorganic Light Emitting Diode Array for High Resolution Optogenetic Devices

Authors: Keundong Lee, Dongha Yoo, Youngbin Tchoe, Gyu-Chul Yi

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Light emitting diode (LED) array is an ideal optical stimulation tool for optogenetics, which controls inhibition and excitation of specific neurons with light-sensitive ion channels or pumps. Although a fiber-optic cable with an external light source, either a laser or LED mechanically connected to the end of the fiber-optic cable has widely been used for illumination on neural tissue, a new approach to use micro LEDs (µLEDs) has recently been demonstrated. The LEDs can be placed directly either on the cortical surface or within the deep brain using a penetrating depth probe. Accordingly, this method would not need a permanent opening in the skull if the LEDs are integrated with miniature electrical power source and wireless communication. In addition, multiple color generation from single µLED cell would enable to excite and/or inhibit neurons in localized regions. Here, we demonstrate flexible and color tunable µLEDs for the optogenetic device applications. The flexible and color tunable LEDs was fabricated using multifaceted gallium nitride (GaN) nanorod arrays with GaN nanorods grown on InxGa1−xN/GaN single quantum well structures (SQW) anisotropically formed on the nanorod tips and sidewalls. For various electroluminescence (EL) colors, current injection paths were controlled through a continuous p-GaN layer depending on the applied bias voltage. The electric current was injected through different thickness and composition, thus changing the color of light from red to blue that the LED emits. We believe that the flexible and color tunable µLEDs enable us to control activities of the neuron by emitting various colors from the single µLED cell.

Keywords: light emitting diode, optogenetics, graphene, flexible optoelectronics

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4193 Morphology of Cartographic Words: A Perspective from Chinese Characters

Authors: Xinyu Gong, Zhilin Li, Xintao Liu

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Maps are a means of communication. Cartographic language involves established theories of natural language for understanding maps. “Cartographic words’, or “map symbols”, are crucial elements of cartographic language. Personalized mapping is increasingly popular, with growing demands for customized map-making by the general public. Automated symbol-making and customization play a key role in personalized mapping. However, formal representations for the automated construction of map symbols are still lacking. In natural language, the process of word and sentence construction can be formalized. Through the analogy between natural language and graphical language, formal representations of natural language construction can be used as a reference for constructing cartographic language. We selected Chinese character structures (i.e., S

Keywords: personalized mapping, Chinese character, cartographic language, map symbols

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4192 Eudesmane-Type Sesquiterpenes from Laggera alata Inhibiting Angiogenesis

Authors: Liang Ning, Chung Hau Yin

Abstract:

Angiogenesis is the process of new blood vessel development. It has been recognized as a therapeutic target for blocking cancer growth four decades ago. Vascular sprouting is initiated by pro-angiogenic factors. Vascular endothelial cell growth factor (VEGF) plays a central role in angiogenic initiation, many patients with cancer or ocular neovascularization have been benefited from anti-VEGF therapy. Emerging approaches impacting in the later stages of vessel remodeling and maturation are expected to improve clinical efficacy. TIE receptor as well as the corresponding angiopoietin ligands, were identified as another endothelial cell specific receptor tyrosine kinase signaling system. Much efforts were made to reduce the activity of angiopoietin-TIE receptor axis. Two eudesmane-type sesquiterpenes from laggera alata, namely, 15-dihydrocostic acid and ilicic acid were found with strong anti-angiogenic properties in zebrafish model. Meanwhile, the mRNA expression levels of VEGFR2 and TIE2 pathway related genes were down-regulated in the sesquiterpenes treated zebrafish embryos. Besides, in human umbilical vein endothelial cells (HUVECs), the sesquiterpenes have the ability to inhibit VEGF-induced HUVECs proliferation and migration at non-toxic concentration. Moreover, angiopoietin-2 induced TIE2 phosphorylation was inhibited by the sesquiterpenes, the inhibitory effect was detected in angiopoietin-1 induced HUVECs proliferation as well. Thus, we hypothesized the anti-angiogenic activity of the compounds may via the inhibition of VEGF and TIE2 related pathways. How the compounds come into play as the pathways inhibitors need to be evaluated in the future.

Keywords: Laggera alata, eudesmane-type sesquiterpene, anti-angiogenesis, VEGF, angiopoietin, TIE2

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4191 Graphene-Oxide-Supported Coal-Layered Double Hydroxides: Synthesis and Characterizations

Authors: Shaeel A. Al Thabaiti, Sulaiman N. Basahel, Salem M. Bawaked, Mohamed Mokhtar

Abstract:

Nanosheets for cobalt-layered double hydroxide (Co-Al-LDH)/GO were successfully synthesized with different Co:M g:Al ratios (0:3:1, 1.5:1.5:1, and 3:0:1). The layered double hydroxide structure and morphology were determined using x-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), and scanning electron microscopy (SEM). Temperature prgrammed reduction (TPR) of Co-Al-LDH showed reduction peaks at lower temperature which indicates the ease reducibility of this particular sample. The thermal behaviour was studied using thermal graviemetric technique (TG), and the BET-surface area was determined using N2 physisorption at -196°C. The C-C coupling reaction was carried out over all the investigated catalysts. The Mg–Al LDH catalyst without Co ions is inactive, but the isomorphic substitution of Mg by Co ions (Co:Mg:Al = 1.5:1.5:1) in the cationic sheet resulted in 88% conversion of iodobenzene under reflux. LDH/GO hybrid is up to 2 times higher activity than for the unsupported LDH.

Keywords: adsorption, co-precipitation, graphene oxide, layer double hydroxide

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4190 Effect of Oxytocin on Cytosolic Calcium Concentration of Alpha and Beta Cells in Pancreas

Authors: Rauza Sukma Rita, Katsuya Dezaki, Yuko Maejima, Toshihiko Yada

Abstract:

Oxytocin is a nine-amino acid peptide synthesized in the paraventricular nucleus (PVN) and supraoptic nucleus (SON) of the hypothalamus. Oxytocin promotes contraction of the uterus during birth and milk ejection during breast feeding. Although oxytocin receptors are found predominantly in the breasts and uterus of females, many tissues and organs express oxytocin receptors, including the pituitary, heart, kidney, thymus, vascular endothelium, adipocytes, osteoblasts, adrenal gland, pancreatic islets, and many cell lines. On the other hand, in pancreatic islets, oxytocin receptors are expressed in both α-cells and β-cells with stronger expression in α- cells. However, to our knowledge there are no reports yet about the effect of oxytocin on cytosolic calcium reaction on α and β-cell. This study aims to investigate the effect of oxytocin on α-cells and β-cells and its oscillation pattern. Islet of Langerhans from wild type mice were isolated by collagenase digestion. Isolated and dissociated single cells either α-cells or β-cells on coverslips were mounted in an open chamber and superfused in HKRB. Cytosolic concentration ([Ca2+]i) in single cells were measured by fura-2 microfluorimetry. After measurement of [Ca2+]i, α-cells were identified by subsequent immunocytochemical staining using an anti-glucagon antiserum. In β-cells, the [Ca2+]i increase in response to oxytocin was observed only under 8.3 mM glucose condition, whereas in α-cells, [Ca2+]i an increase induced by oxytocin was observed in both 2.8 mM and 8.3 mM glucose. The oscillation incidence was induced more frequently in β-cells compared to α-cells. In conclusion, the present study demonstrated that oxytocin directly interacts with both α-cells and β-cells and induces increase of [Ca2+]i and its specific patterns.

Keywords: α-cells, β-cells, cytosolic calcium concentration, oscillation, oxytocin

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4189 Experimental and Theoratical Methods to Increase Core Damping for Sandwitch Cantilever Beam

Authors: Iyd Eqqab Maree, Moouyad Ibrahim Abbood

Abstract:

The purpose behind this study is to predict damping effect for steel cantilever beam by using two methods of passive viscoelastic constrained layer damping. First method is Matlab Program, this method depend on the Ross, Kerwin and Unger (RKU) model for passive viscoelastic damping. Second method is experimental lab (frequency domain method), in this method used the half-power bandwidth method and can be used to determine the system loss factors for damped steel cantilever beam. The RKU method has been applied to a cantilever beam because beam is a major part of a structure and this prediction may further leads to utilize for different kinds of structural application according to design requirements in many industries. In this method of damping a simple cantilever beam is treated by making sandwich structure to make the beam damp, and this is usually done by using viscoelastic material as a core to ensure the damping effect. The use of viscoelastic layers constrained between elastic layers is known to be effective for damping of flexural vibrations of structures over a wide range of frequencies. The energy dissipated in these arrangements is due to shear deformation in the viscoelastic layers, which occurs due to flexural vibration of the structures. The theory of dynamic stability of elastic systems deals with the study of vibrations induced by pulsating loads that are parametric with respect to certain forms of deformation. There is a very good agreement of the experimental results with the theoretical findings. The main ideas of this thesis are to find the transition region for damped steel cantilever beam (4mm and 8mm thickness) from experimental lab and theoretical prediction (Matlab R2011a). Experimentally and theoretically proved that the transition region for two specimens occurs at modal frequency between mode 1 and mode 2, which give the best damping, maximum loss factor and maximum damping ratio, thus this type of viscoelastic material core (3M468) is very appropriate to use in automotive industry and in any mechanical application has modal frequency eventuate between mode 1 and mode 2.

Keywords: 3M-468 material core, loss factor and frequency, domain method, bioinformatics, biomedicine, MATLAB

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4188 Structural Strength Evaluation and Wear Prediction of Double Helix Steel Wire Ropes for Heavy Machinery

Authors: Krunal Thakar

Abstract:

Wire ropes combine high tensile strength and flexibility as compared to other general steel products. They are used in various application areas such as cranes, mining, elevators, bridges, cable cars, etc. The earliest reported use of wire ropes was for mining hoist application in 1830s. Over the period, there have been substantial advancement in the design of wire ropes for various application areas. Under operational conditions, wire ropes are subjected to varying tensile loads and bending loads resulting in material wear and eventual structural failure due to fretting fatigue. The conventional inspection methods to determine wire failure is only limited to outer wires of rope. However, till date, there is no effective mathematical model to examine the inter wire contact forces and wear characteristics. The scope of this paper is to present a computational simulation technique to evaluate inter wire contact forces and wear, which are in many cases responsible for rope failure. Two different type of ropes, IWRC-6xFi(29) and U3xSeS(48) were taken for structural strength evaluation and wear prediction. Both ropes have a double helix twisted wire profile as per JIS standards and are mainly used in cranes. CAD models of both ropes were developed in general purpose design software using in house developed formulation to generate double helix profile. Numerical simulation was done under two different load cases (a) Axial Tension and (b) Bending over Sheave. Different parameters such as stresses, contact forces, wear depth, load-elongation, etc., were investigated and compared between both ropes. Numerical simulation method facilitates the detailed investigation of inter wire contact and wear characteristics. In addition, various selection parameters like sheave diameter, rope diameter, helix angle, swaging, maximum load carrying capacity, etc., can be quickly analyzed.

Keywords: steel wire ropes, numerical simulation, material wear, structural strength, axial tension, bending over sheave

Procedia PDF Downloads 152
4187 Phylogenetic Differential Separation of Environmental Samples

Authors: Amber C. W. Vandepoele, Michael A. Marciano

Abstract:

Biological analyses frequently focus on single organisms, however many times, the biological sample consists of more than the target organism; for example, human microbiome research targets bacterial DNA, yet most samples consist largely of human DNA. Therefore, there would be an advantage to removing these contaminating organisms. Conversely, some analyses focus on a single organism but would greatly benefit from the additional information regarding the other organismal components of the sample. Forensic analysis is one such example, wherein most forensic casework, human DNA is targeted; however, it typically exists in complex non-pristine sample substrates such as soil or unclean surfaces. These complex samples are commonly comprised of not just human tissue but also microbial and plant life, where these organisms may help gain more forensically relevant information about a specific location or interaction. This project aims to optimize a ‘phylogenetic’ differential extraction method that will separate mammalian, bacterial and plant cells in a mixed sample. This is accomplished through the use of size exclusion separation, whereby the different cell types are separated through multiple filtrations using 5 μm filters. The components are then lysed via differential enzymatic sensitivities among the cells and extracted with minimal contribution from the preceding component. This extraction method will then allow complex DNA samples to be more easily interpreted through non-targeting sequencing since the data will not be skewed toward the smaller and usually more numerous bacterial DNAs. This research project has demonstrated that this ‘phylogenetic’ differential extraction method successfully separated the epithelial and bacterial cells from each other with minimal cell loss. We will take this one step further, showing that when adding the plant cells into the mixture, they will be separated and extracted from the sample. Research is ongoing, and results are pending.

Keywords: DNA isolation, geolocation, non-human, phylogenetic separation

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4186 Modeling and Prediction of Hot Deformation Behavior of IN718

Authors: M. Azarbarmas, J. M. Cabrera, J. Calvo, M. Aghaie-Khafri

Abstract:

The modeling of hot deformation behavior for unseen conditions is important in metal-forming. In this study, the hot deformation of IN718 has been characterized in the temperature range 950-1100 and strain rate range 0.001-0.1 s-1 using hot compression tests. All stress-strain curves showed the occurrence of dynamic recrystallization. These curves were implemented quantitatively in mathematics, and then constitutive equation indicating the relationship between the flow stress and hot deformation parameters was obtained successfully.

Keywords: compression test, constitutive equation, dynamic recrystallization, hot working

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4185 The Role of Bone Marrow Stem Cells Transplantation in the Repair of Damaged Inner Ear in Albino Rats

Authors: Ahmed Gaber Abdel Raheem, Nashwa Ahmed Mohamed

Abstract:

Introduction: Sensorineural hearing loss (SNHL) is largely caused by the degeneration of the cochlea. Therapeutic options for SNHL are limited to hearing aids and cochlear implants. The cell transplantation approach to the regeneration of hair cells has gained considerable attention because stem cells are believed to accumulate in the damaged sites and have the potential for the repair of damaged tissues. The aim of the work: was to assess the use of bone marrow transplantation in repair of damaged inner ear hair cells in rats after the damage had been inflicted by Amikacin injection. Material and Methods: Thirty albino rats were used in this study. They were divided into three groups. Each group ten rats. Group I: used as control. Group II: Were given Amikacin- intratympanic injection till complete loss of hearing function. This could be assessed by Distortion product Otoacoustic Emission (DPOAEs) and / or auditory brain stem evoked potential (ABR). GroupIII: were given intra-peritoneal injection of bone marrow stem cell after complete loss of hearing caused by Amikacin. Clinical assessment was done using DPOAEs and / or auditory brain stem evoked potential (ABR), before and after bone marrow injection. Histological assessment of the inner ear was done by light and electron microscope. Also, Detection of stem cells in the inner ear by immunohistochemistry. Results: Histological examination of the specimens showed promising improvement in the structure of cochlea that may be responsible for the improvement of hearing function in rats detected by DPOAEs and / or ABR. Conclusion: Bone marrow stem cells transplantation might be useful for the treatment of SNHL.

Keywords: amikacin, hair cells, sensorineural hearing loss, stem cells

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4184 Prediction of Crack Propagation in Bonded Joints Using Fracture Mechanics

Authors: Reza Hedayati, Meysam Jahanbakhshi

Abstract:

In this work, Fracture Mechanics is used to predict crack propagation in the adhesive jointing aluminum and composite plates. Three types of loadings and two types of glass-epoxy composite sequences: [0/90]2s and [0/45/-45/90]s are considered for the composite plate. Therefore 2*3=6 cases are considered and their results are compared. The debonding initiation load, complete debonding load, crack face profile and load-displacement diagram have been compared for the six cases.

Keywords: fracture, adhesive joint, debonding, APDL, LEFM

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4183 Monitoring of Latent Tree Mortality after Forest Fires: A Biosensor Approach

Authors: Alessio Giovannelli, Claudia Cocozza, Enrico Marchi, Valerio Giorgio Muzzini, Eleftherios Touloupakis, Raffaella Margherita Zampieri

Abstract:

In Mediterranean countries, forest fires are recurrent events that need to be considered as a central component of regional and global forest management strategies and biodiversity restoration programmes. The response of tree function to fire damage can vary widely, also taking into account species, season, age of the tree, etc. Trees that survive fire may have different levels of physiological functionality, which may result in reduced growth or increased susceptibility to delayed mortality. An approach to assessing irreversible physiological injury in trees could help to inform management decisions at burned sites for biodiversity restoration, environmental safety and understanding of ecosystem functional adaptations. Physiological proxies for latent tree mortality, such as cambial cell death, reduced or absent starch and soluble sugar content in C sinks, and ethanol accumulation in the phloem, are considered proxies for cell death. However, their determination requires time-consuming laboratory protocols, making the approach unfeasible as a practical option in the field, but recent findings have shown that biosensors could be usefully applied to overcome these limitations. The study will focus on the development of amperometric biosensors capable of detecting a few target molecules in the phloem and xylem (such as ethanol and glucose) that have recently been identified as proxies for latent tree mortality. The results of a specific experiment on a stand of Pinus pinaster subjected to prescribed fire are reported.

Keywords: enzymes, glucose, ethanol, prescribed fires

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4182 Texture Analysis of Grayscale Co-Occurrence Matrix on Mammographic Indexed Image

Authors: S. Sushma, S. Balasubramanian, K. C. Latha

Abstract:

The mammographic image of breast cancer compressed and synthesized to get co-efficient values which will be converted (5x5) matrix to get ROI image where we get the highest value of effected region and with the same ideology the technique has been extended to differentiate between Calcification and normal cell image using mean value derived from 5x5 matrix values

Keywords: texture analysis, mammographic image, partitioned gray scale co-oocurance matrix, co-efficient

Procedia PDF Downloads 534
4181 Photocatalytic Degradation of Organic Polluant Reacting with Tungstates: Role of Microstructure and Size Effect on Oxidation Kinetics

Authors: A. Taoufyq, B. Bakiz, A. Benlhachemi, L. Patout, D. V. Chokouadeua, F. Guinneton, G. Nolibe, A. Lyoussi, J-R. Gavarri

Abstract:

Currently, the photo catalytic reactions occurring under solar illumination have attracted worldwide attentions due to a tremendous set of environmental problems. Taking the sunlight into account, it is indispensable to develop highly effective visible-light-driver photo catalysts. Nano structured materials such as MxM’1-xWO6 system are widely studied due to its interesting piezoelectric, dielectric and catalytic properties. These materials can be used in photo catalysis technique for environmental applications, such as waste water treatments. The aim of this study was to investigate the photo catalytic activity of polycrystalline phases of bismuth tungstate of formula Bi2WO6. Polycrystalline samples were elaborated using a coprecipitation technique followed by a calcination process at different temperatures (300, 400, 600 and 900°C). The obtained polycrystalline phases have been characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Crystal cell parameters and cell volume depend on elaboration temperature. High-resolution electron microscopy images and image simulations, associated with X-ray diffraction data, allowed confirming the lattices and space groups Pca21. The photo catalytic activity of the as-prepared samples was studied by irradiating aqueous solutions of Rhodamine B, associated with Bi2WO6 additives having variable crystallite sizes. The photo catalytic activity of such bismuth tungstates increased as the crystallite sizes decreased. The high specific area of the photo catalytic particles obtained at 300°C seems to condition the degradation kinetics of RhB.

Keywords: Bismuth tungstate, crystallite sizes, electron microscopy, photocatalytic activity, X-ray diffraction.

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4180 Green Synthesis of Silver Nanoparticles Using Echinacea Flower Extract and Characterization

Authors: Masood Hussain, Erol Pehlivan, Ahmet Avci, Ecem Guder

Abstract:

Green synthesis of silver nanoparticles (AgNPs) was carried out by using echinacea flower extract as reducing/protecting agent. The effects of various operating parameters and additives on the dimensions such as stirring rate, temperature, pH of the solution, the amount of extract and concentration of silver nitrate were optimized in order to achieve monodispersed spherical and small size echinacea protected silver nanoparticles (echinacea-AgNPs) through biosynthetic method. The surface roughness and topography of synthesized metal nanoparticles were confirmed by using Atomic Force Microscopy (AFM). High-Resolution Transmission Electron Microscopic (HRTEM) results elaborated the formation of uniformly distributed Echinacea protected AgNPs (Echinacea-AgNPs) having an average size of 30.2±2nm.

Keywords: Echinacea flower extract, green synthesis, silver nanoparticles, morphology

Procedia PDF Downloads 422
4179 Application of Flow Cytometry for Detection of Influence of Abiotic Stress on Plants

Authors: Dace Grauda, Inta Belogrudova, Alexei Katashev, Linda Lancere, Isaak Rashal

Abstract:

The goal of study was the elaboration of easy applicable flow cytometry method for detection of influence of abiotic stress factors on plants, which could be useful for detection of environmental stresses in urban areas. The lime tree Tillia vulgaris H. is a popular tree species used for urban landscaping in Europe and is one of the main species of street greenery in Riga, Latvia. Tree decline and low vitality has observed in the central part of Riga. For this reason lime trees were select as a model object for the investigation. During the period of end of June and beginning of July 12 samples from different urban environment locations as well as plant material from a greenhouse were collected. BD FACSJazz® cell sorter (BD Biosciences, USA) with flow cytometer function was used to test viability of plant cells. The method was based on changes of relative fluorescence intensity of cells in blue laser (488 nm) after influence of stress factors. SpheroTM rainbow calibration particles (3.0–3.4 μm, BD Biosciences, USA) in phosphate buffered saline (PBS) were used for calibration of flow cytometer. BD PharmingenTM PBS (BD Biosciences, USA) was used for flow cytometry assays. The mean fluorescence intensity information from the purified cell suspension samples was recorded. Preliminary, multiple gate sizes and shapes were tested to find one with the lowest CV. It was found that low CV can be obtained if only the densest part of plant cells forward scatter/side scatter profile is analysed because in this case plant cells are most similar in size and shape. The young pollen cells in one nucleus stage were found as the best for detection of influence of abiotic stress. For experiments only fresh plant material was used– the buds of Tillia vulgaris with diameter 2 mm. For the cell suspension (in vitro culture) establishment modified protocol of microspore culture was applied. The cells were suspended in the MS (Murashige and Skoog) medium. For imitation of dust of urban area SiO2 nanoparticles with concentration 0.001 g/ml were dissolved in distilled water. Into 10 ml of cell suspension 1 ml of SiO2 nanoparticles suspension was added, then cells were incubated in speed shaking regime for 1 and 3 hours. As a stress factor the irradiation of cells for 20 min by UV was used (Hamamatsu light source L9566-02A, L10852 lamp, A10014-50-0110), maximum relative intensity (100%) at 365 nm and at ~310 nm (75%). Before UV irradiation the suspension of cells were placed onto a thin layer on a filter paper disk (diameter 45 mm) in a Petri dish with solid MS media. Cells without treatment were used as a control. Experiments were performed at room temperature (23-25 °C). Using flow cytometer BS FACS Software cells plot was created to determine the densest part, which was later gated using oval-shaped gate. Gate included from 95 to 99% of all cells. To determine relative fluorescence of cells logarithmic fluorescence scale in arbitrary fluorescence units were used. 3x103 gated cells were analysed from the each sample. The significant differences were found among relative fluorescence of cells from different trees after treatment with SiO2 nanoparticles and UV irradiation in comparison with the control.

Keywords: flow cytometry, fluorescence, SiO2 nanoparticles, UV irradiation

Procedia PDF Downloads 413