Search results for: adipose derived stem cells
4384 Inductions of CaC₂ on Sperm Morphology and Viability of the Albino Mice (Mus musculus)
Authors: Dike H. Ogbuagu, Etsede J. Oritsematosan
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This work investigated possible inductions of CaC₂, often misused by fruit vendors to stimulate artificial ripening, on mammalian sperm morphology and viability. Thirty isogenic strains of male albino mice, Mus musculus (age≈ 8weeks; weight= 32.5±2.0g) were acclimatized (ambient temperature 28.0±1.0°C) for 2 weeks and fed standard growers mash and water ad libutum. They were later exposed to graded toxicant concentrations (w/w) of 2.5000, 1.2500, 0.6250, and 0.3125% in 4 cages. A control cage was also established. After 5 weeks, 3 animals from each cage were sacrificed by cervical dislocation and the cauda epididymis excised. Sperm morphology and viability were determined by microscopic procedures. The ANOVA, means plots, Student’s t-test and variation plots were used to analyze data. The common abnormalities observed included Double Head, Pin Head, Knobbed Head, No Tail and With Hook. The higher toxicant concentrations induced significantly lower body weights [F(829.899) ˃ Fcrit(4.19)] and more abnormalities [F(26.52) ˃ Fcrit(4.00)] at P˂0.05. Sperm cells in the control setup were significantly more viable than those in the 0.625% (t=0.005) and 2.500% toxicant doses (t=0.018) at the 95% confidence limit. CaC₂ appeared to induced morphological abnormalities and reduced viability in sperm cells of M. musculus.Keywords: artificial ripening, calcium carbide, fruit vendors, sperm morphology, sperm viability
Procedia PDF Downloads 2234383 Disaggregation of Coarser Resolution Radiometer Derived Soil Moisture to Finer Scales
Authors: Gurjeet Singh, Rabindra K. Panda
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Soil moisture is a key hydrologic state variable and is intrinsically linked to the Earth's water, climate and carbon cycles. On ecological point of view, the soil moisture is a fundamental natural resource providing the transpirable water for plants. Soil moisture varies both temporally and spatially due to spatiotemporal variation in rainfall, vegetation cover, soil properties and topography. Satellite derived soil moisture provides spatio-temporal extensive data. However, the spatial resolution of a typical satellite (L-band radiometry) is of the order of tens of kilometers, which is not good enough for developing efficient agricultural water management schemes at the field scale. In the present study, the soil moisture from radiometer data has been disaggregated using blending approach to achieve higher resolution soil moisture data. The radiometer estimates of soil moisture at a 40 km resolution have been disaggregated to 10 km, 5 km and 1 km resolutions. The disaggregated soil moisture was compared with the observed data, consisting of continuous sensor based soil moisture profile measurements, at three monitoring sites and extensive spatial near-surface soil moisture measurements, concurrent with satellite monitoring in the 500 km2 study watershed in the Eastern India. The estimated soil moisture status at different spatial scales can help in developing efficient agricultural water management schemes to increase the crop production and water use efficiency.Keywords: disaggregation, eastern India, radiometers, soil moisture, water use efficiency
Procedia PDF Downloads 2784382 Digital Joint Equivalent Channel Hybrid Precoding for Millimeterwave Massive Multiple Input Multiple Output Systems
Authors: Linyu Wang, Mingjun Zhu, Jianhong Xiang, Hanyu Jiang
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Aiming at the problem that the spectral efficiency of hybrid precoding (HP) is too low in the current millimeter wave (mmWave) massive multiple input multiple output (MIMO) system, this paper proposes a digital joint equivalent channel hybrid precoding algorithm, which is based on the introduction of digital encoding matrix iteration. First, the objective function is expanded to obtain the relation equation, and the pseudo-inverse iterative function of the analog encoder is derived by using the pseudo-inverse method, which solves the problem of greatly increasing the amount of computation caused by the lack of rank of the digital encoding matrix and reduces the overall complexity of hybrid precoding. Secondly, the analog coding matrix and the millimeter-wave sparse channel matrix are combined into an equivalent channel, and then the equivalent channel is subjected to Singular Value Decomposition (SVD) to obtain a digital coding matrix, and then the derived pseudo-inverse iterative function is used to iteratively regenerate the simulated encoding matrix. The simulation results show that the proposed algorithm improves the system spectral efficiency by 10~20%compared with other algorithms and the stability is also improved.Keywords: mmWave, massive MIMO, hybrid precoding, singular value decompositing, equivalent channel
Procedia PDF Downloads 1044381 Optimal Pricing Based on Real Estate Demand Data
Authors: Vanessa Kummer, Maik Meusel
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Real estate demand estimates are typically derived from transaction data. However, in regions with excess demand, transactions are driven by supply and therefore do not indicate what people are actually looking for. To estimate the demand for housing in Switzerland, search subscriptions from all important Swiss real estate platforms are used. These data do, however, suffer from missing information—for example, many users do not specify how many rooms they would like or what price they would be willing to pay. In economic analyses, it is often the case that only complete data is used. Usually, however, the proportion of complete data is rather small which leads to most information being neglected. Also, the data might have a strong distortion if it is complete. In addition, the reason that data is missing might itself also contain information, which is however ignored with that approach. An interesting issue is, therefore, if for economic analyses such as the one at hand, there is an added value by using the whole data set with the imputed missing values compared to using the usually small percentage of complete data (baseline). Also, it is interesting to see how different algorithms affect that result. The imputation of the missing data is done using unsupervised learning. Out of the numerous unsupervised learning approaches, the most common ones, such as clustering, principal component analysis, or neural networks techniques are applied. By training the model iteratively on the imputed data and, thereby, including the information of all data into the model, the distortion of the first training set—the complete data—vanishes. In a next step, the performances of the algorithms are measured. This is done by randomly creating missing values in subsets of the data, estimating those values with the relevant algorithms and several parameter combinations, and comparing the estimates to the actual data. After having found the optimal parameter set for each algorithm, the missing values are being imputed. Using the resulting data sets, the next step is to estimate the willingness to pay for real estate. This is done by fitting price distributions for real estate properties with certain characteristics, such as the region or the number of rooms. Based on these distributions, survival functions are computed to obtain the functional relationship between characteristics and selling probabilities. Comparing the survival functions shows that estimates which are based on imputed data sets do not differ significantly from each other; however, the demand estimate that is derived from the baseline data does. This indicates that the baseline data set does not include all available information and is therefore not representative for the entire sample. Also, demand estimates derived from the whole data set are much more accurate than the baseline estimation. Thus, in order to obtain optimal results, it is important to make use of all available data, even though it involves additional procedures such as data imputation.Keywords: demand estimate, missing-data imputation, real estate, unsupervised learning
Procedia PDF Downloads 2924380 Effect of Probiotics and Vitamin B on Plasma Interferon-Gamma and Interleukin-6 Levels in Active Pulmonary Tuberculosis
Authors: Yulistiani Yulistiani, Zamrotul Izzah, Lintang Bismantara, Wenny Putri Nilamsari, Arif Bachtiar, Budi Suprapti
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Interferon-gamma (IFN-γ) and interleukin-6 (IL-6) are pro-inflammatory cytokines, which have the protective immune response against Tuberculosis (TB). Indeed, pro-inflammatory cytokines Mycobacterium tuberculosis antigen-specific CD4+ and CD8+ T cells and NK cells increase the level of production of IFN-γ, a cytokine critical for augmenting the microbicidal activity of phagocytes. On the other hand, M. tuberculosis reduces the effects of IFN-γ by inhibiting the transcription of IFN-γ- responsive genes and by inducing the secretion of IL-6, which inhibits IFN-γ signaling. Probiotics Lactobacillus sp. and Bifidobacterium sp. were known to increase IFN-γ production in vivo, while vitamin B1, B6, and B12 worked on macrophages and releasing cytokines. Therefore, the present study was to evaluate the effect of probiotics and vitamin B supplement on changes of plasma cytokine levels in active pulmonary TB. From October to November 2016, twelve M. tuberculosis-infected patients starting anti-TB drugs were recruited, then divided into two groups. Seven patients were given a combination of probiotics and vitamin B, while five patients were in the control group. Plasma IFN-γ and IL-6 levels were measured by the ELISA kit before and a month after treatment. IFN-γ levels raised in four patients receiving the supplement (P = 0.743), while IL-6 increased in three patients in this group until day 30 of treatment (P = 0.298). Taken together, these results show the promising effect of probiotics and vitamin B on stimulation of IFN-γ and IL-6 production during intensive therapy of TB.Keywords: interferon-gamma, interleukin-6, probiotic, tuberculosis
Procedia PDF Downloads 3524379 The Regulation of the Pro-inflammatory Cytokine Interleukin 6 (IL6) by Epstein-Barr Virus (EBV)
Authors: Liu Xiaohan
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Epstein–Barr virus (EBV) is a human herpesvirus and is closely related to many malignancies of lymphocyte and epithelial origins, such as gastric cancer, Burkitt’s lymphoma, and nasopharyngeal carcinoma (NPC). NPC is a malignant epithelial tumor which is 100% associated with EBV latent infection. Most of the NPC cases are densely populated in southern China, especially in Guangdong and Hong Kong. To our knowledge, overexpression of pro-inflammatory cytokines may result in a loss of balance of the immune system and cause damage to human bodies. Interleukin-6 (IL6) is a pro-inflammatory cytokine which plays an important role in tumor progression. In addition, gene expression is regulated by both transcriptional and post-transcriptional pathways, while post-transcriptional regulation is an important mechanism to modulate the mature mRNA level in mammalian cells. AU-rich element binding factor 1 (AUF1)/heterogeneous nuclear RNP D (hnRNP D) is known for its function in destabilizing mRNAs, including cytokines and cell cycle regulators. Previous studies have found that overexpression of hnRNP D would lead to tumorigenesis. In this project, our aim is to determine the role played by hnRNP D in EBV-infected cells and how our anti-EBV agents can affect the function of hnRNP D. The results of this study will provide a new insight into how the pro-inflammatory cytokine expression can be regulated by EBV.Keywords: interleukin 6 (IL6), epstein-barr virus (EBV), nasopharyngeal carcinoma (NPC, epstein-barr nuclear antigen-1 (EBNA1)
Procedia PDF Downloads 664378 Impact of Behavioral Biases on Indian Investors: Case Analysis of a Mutual Fund Investment Company
Authors: Priyal Motwani, Garvit Goel
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In this study, we have studied and analysed the transaction data of investors of a mutual fund investment company based in India. Based on the data available, we have identified the top four biases that affect the investors of the emerging market economies through regression analysis and three uniquely defined ratios. We found that the four most prominent biases that affected the investment making decisions in India are– Chauffer Knowledge, investors tend to make ambitious decisions about sectors they know little about; Bandwagon effect – the response of the market indices to macroeconomic events are more profound and seem to last longer compared to western markets; base-rate neglect – judgement about stocks are too much based on the most recent development ignoring the long-term fundamentals of the stock; availability bias – lack of proper communication channels of market information lead people to be too reliant on limited information they already have. After segregating the investors into six groups, the results have further been studied to identify a correlation among the demographics, gender and unique cultural identity of the derived groups and the corresponding prevalent biases. On the basis of the results obtained from the derived groups, our study recommends six methods, specific to each group, to educate the investors about the prevalent biases and their role in investment decision making.Keywords: Bandwagon effect, behavioural biases, Chauffeur knowledge, demographics, investor literacy, mutual funds
Procedia PDF Downloads 2334377 Hybrid Polymer Microfluidic Platform for Studying Endothelial Cell Response to Micro Mechanical Environment
Authors: Mitesh Rathod, Jungho Ahn, Noo Li Jeon, Junghoon Lee
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Endothelial cells respond to cues from both biochemical as well as micro mechanical environment. Significant effort has been directed to understand the effects of biochemical signaling, however, relatively little is known about regulation of endothelial cell biology by the micro mechanical environment. Numerous studies have been performed to understand how physical forces regulate endothelial cell behavior. In this regard, past studies have majorly focused on exploring how fluid shear stress governs endothelial cell behavior. Parallel plate flow chambers and rectangular microchannels are routinely employed for applying fluid shear force on endothelial cells. However, these studies fall short in mimicking the in vivo like micro environment from topological aspects. Few studies have only used circular microchannels to replicate in vivo like condition. Seldom efforts have been directed to elucidate the combined effect of topology, substrate rigidity and fluid shear stress on endothelial cell response. In this regard, we demonstrate a facile fabrication process to develop a hybrid polydimethylsiloxane microfluidic platform to study endothelial cell biology. On a single chip microchannels with different cross sections i.e., circular, rectangular and square have been fabricated. In addition, our fabrication approach allows variation in the substrate rigidity along the channel length. Two different variants of polydimethylsiloxane, namely Sylgard 184 and Sylgard 527, were utilized to achieve the variation in rigidity. Moreover, our approach also enables in creating Y bifurcation circular microchannels. Our microfluidic platform thus facilitates for conducting studies pertaining to endothelial cell morphology with respect to change in topology, substrate rigidity and fluid flow on a single chip. The hybrid platform was tested by culturing Human Umbilical Vein Endothelial Cells in circular microchannels with varying substrate rigidity, and exposed to fluid shear stress of 12 dynes/cm² and static conditions. Results indicate the cell area response to flow induced shear stress was governed by the underlying substrate mechanics.Keywords: hybrid, microfluidic platform, PDMS, shear flow, substrate rigidity
Procedia PDF Downloads 2794376 A Methodology for the Synthesis of Multi-Processors
Authors: Hamid Yasinian
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Random epistemologies and hash tables have garnered minimal interest from both security experts and experts in the last several years. In fact, few information theorists would disagree with the evaluation of expert systems. In our research, we discover how flip-flop gates can be applied to the study of superpages. Though such a hypothesis at first glance seems perverse, it is derived from known results.Keywords: synthesis, multi-processors, interactive model, moor’s law
Procedia PDF Downloads 4404375 DNAJB6 Chaperone Prevents the Aggregation of Intracellular but not Extracellular Aβ Peptides Associated with Alzheimer’s Disease
Authors: Rasha M. Hussein, Reem M. Hashem, Laila A. Rashed
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Alzheimer’s disease is the most common dementia disease in the elderly. It is characterized by the accumulation of extracellular amyloid β (Aβ) peptides and intracellular hyper-phosphorylated tau protein. In addition, recent evidence indicates that accumulation of intracellular amyloid β peptides may play a role in Alzheimer’s disease pathogenesis. This suggests that intracellular Heat Shock Proteins (HSP) that maintain the protein quality control in the cell might be potential candidates for disease amelioration. DNAJB6, a member of DNAJ family of HSP, effectively prevented the aggregation of poly glutamines stretches associated with Huntington’s disease both in vitro and in cells. In addition, DNAJB6 was found recently to delay the aggregation of Aβ42 peptides in vitro. In the present study, we investigated the ability of DNAJB6 to prevent the aggregation of both intracellular and extracellular Aβ peptides using transfection of HEK293 cells with Aβ-GFP and recombinant Aβ42 peptides respectively. We performed western blotting and immunofluorescence techniques. We found that DNAJB6 can prevent Aβ-GFP aggregation, but not the seeded aggregation initiated by extracellular Aβ peptides. Moreover, DNAJB6 required interaction with HSP70 to prevent the aggregation of Aβ-GFP protein and its J-domain was essential for this anti-aggregation activity. Interestingly, overexpression of other DNAJ proteins as well as HSPB1 suppressed Aβ-GFP aggregation efficiently. Our findings suggest that DNAJB6 is a promising candidate for the inhibition of Aβ-GFP mediated aggregation through a canonical HSP70 dependent mechanism.Keywords: Aβ, Alzheimer’s disease, chaperone, DNAJB6, aggregation
Procedia PDF Downloads 5194374 Enhancement in Antimicrobial and Antioxidant Activity of Cuminum cyminum L. through Niosome Nanocarries
Authors: Fatemeh Haghiralsadat, Mohadese Hashemi, Elham Akhoundi Kharanaghi, Mojgan Yazdani, Mahboobe Sharafodini, Omid Javani
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Niosomes are colloidal particles formed from the self-assembly of non-ionic surfactants in aqueous medium resulting in closed bilayer structures. As a consequence of this hydrophilic and hydrophobic structure, niosomes have the capacity to entrap compounds of different solubilities. Niosomes are promising vehicle for drug delivery which protect sensitive drugs and improve the therapeutic index of drugs by restricting their action to target cells. Essential oils are complex mixtures of volatile compounds such as terpenoids, phenol-derived aromatic components that have been used for many biological properties including bactericidal, fungicidal, insecticidal, antioxidant, anti-tyrosinase and other medicinal properties. Encapsulation of essential oils in niosomes can be an attractive method to overcome their limitation such as volatility, easily decomposition by heat, humidity, light, or oxygen. Cuminum cyminum L. (Cumin) is an aromatic plant included in the Apiaceae family and is used to flavor foods, added to fragrances, and for medical preparations which is indigenous to Egypt, the Mediterranean region, Iran and India. The major components of the Cumin oil were reported as cuminaldehyde, γ -terpinene, β-pinene, p-cymene, p-mentha-1, 3-dien-7-al, and p-mentha-1, 4-dien-7-al which provide the antimicrobial and antioxidant activity. The aim of this work was to formulate Cumin essential oil-loaded niosomes to improve water solubility of natural product and evaluate its physico-chemical features and stability. Cumin oil was obtained through steam distillation using a clevenger-type apparatus and GC/MS was applied to identify the main components of the essential oil. Niosomes were prepared by using thin film hydration method and nanoparticles were characterized for particle size, dispersity index, zeta potential, encapsulation efficiency, in vitro release, and morphology.Keywords: Cuminum cyminum L., Cumin, niosome, essential oil, encapsulation
Procedia PDF Downloads 5234373 A Refined Nonlocal Strain Gradient Theory for Assessing Scaling-Dependent Vibration Behavior of Microbeams
Authors: Xiaobai Li, Li Li, Yujin Hu, Weiming Deng, Zhe Ding
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A size-dependent Euler–Bernoulli beam model, which accounts for nonlocal stress field, strain gradient field and higher order inertia force field, is derived based on the nonlocal strain gradient theory considering velocity gradient effect. The governing equations and boundary conditions are derived both in dimensional and dimensionless form by employed the Hamilton principle. The analytical solutions based on different continuum theories are compared. The effect of higher order inertia terms is extremely significant in high frequency range. It is found that there exists an asymptotic frequency for the proposed beam model, while for the nonlocal strain gradient theory the solutions diverge. The effect of strain gradient field in thickness direction is significant in low frequencies domain and it cannot be neglected when the material strain length scale parameter is considerable with beam thickness. The influence of each of three size effect parameters on the natural frequencies are investigated. The natural frequencies increase with the increasing material strain gradient length scale parameter or decreasing velocity gradient length scale parameter and nonlocal parameter.Keywords: Euler-Bernoulli Beams, free vibration, higher order inertia, Nonlocal Strain Gradient Theory, velocity gradient
Procedia PDF Downloads 2714372 Chemical Characterization, Crystallography and Acute Toxicity Evaluation of Two Boronic-Carbohydrate Adducts
Authors: Héctor González Espinosa, Ricardo Ivan Cordova Chávez, Alejandra Contreras Ramos, Itzia Irene Padilla Martínez, José Guadalupe Trujillo Ferrara, Marvin Antonio Soriano Ursúa
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Boronic acids are able to create diester bonds with carbohydrates because of their hydroxyl groups; in nature, there are some organoborates with these characteristics, such as the calcium fructoborate, formed by the union of two fructose molecules and a boron atom, synthesized by plants. In addition, it has been observed that, in animal cells only the compounds with cis-diol functional groups are capable of linking to boric or boronic acids. The formation of these organoboron compounds could impair the physical and chemical properties of the precursors, even their acute toxicity. In this project, two carbohydrate-derived boron-containing compounds from D-fructose and D-arabinose and phenylboronic acid are analyzed by different spectroscopy techniques such as Raman, Infrared with Fourier Transform Infrared (FT-IR), Nuclear Magnetic Resonance (NMR) and X-ray diffraction crystallography to describe their chemical characteristics. Also, an acute toxicity test was performed to determine their LD50 using the Lorke’s method. It was confirmed by multiple spectra the formation of the adducts by the generation of the diester bonds with a β-D-pyranose of fructose and arabinose. The most prominent findings were the presence of signals corresponding to the formation of new bonds, like the stretching of B-O bonds, or the absence of signals of functional groups like the hydroxyls presented in the reagents used for the synthesis of the adducts. The NMR spectra yielded information about the stereoselectivity in the synthesis reaction, observed by the interaction of the protons and their vicinal atoms in the anomeric and second position carbons; but also, the absence of a racemic mix by the finding of just one signal in the range for the anomeric carbon in the 13C NMR spectra of both adducts. The acute toxicity tests by the Lorke’s method showed that the LD50 value for both compounds is 1265 mg/kg. Those results let us to propose these adducts as highly safe agents for further biological evaluation with medical purposes.Keywords: acute toxicity, adduct, boron, carbohydrate, diester bond
Procedia PDF Downloads 714371 Hydrodeoxygenation of Furfural over RU Sub-Nano Particles Supported on Al₂O₃-SIO₂ Mixed Oxides
Authors: Chaima Zoulikha Tabet Zatla, Nihel Dib, Sumeya Bedrane, Juan Carlos Hernandez Garrido, Redouane Bachir, Miguel Angel Cauqui, Jose Juan Calvino Gamez
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These last year's our planet has witnessed global warming, which is a serious threat to our lives; it has many causes, such as the CO₂ excess in the atmosphere that results from our activity, for the purpose of living in a neater and better environment, working and improving an eco-responsible energy system is a must. Valorization of biomass to produce biofuels is among the most compelling routes to decrease air pollution without considerable modification in current vehicle technology. Effective transformation of lignocellulosic biomass-derived compounds into liquid fuels and value-added chemicals is an economically viable solution. Presently, very competitive technics for the conversion of lignocellulosic biomass into platform chemicals, such as furfural and Hydroxymethylfurfural (HMF), are used. Furfural (C₅H₄O₂) is a major hemi cellulosic biomass-derived platform molecule. In our work, we focus on the valorization of lignocellulosic biomass derivative furfural that is transformed into biofuel through a hydrodeoxygenation reaction in general and involving a catalytic process. In order to get to this point, we are synthesizing and characterizing a series of catalysts with different amounts of Ru (0.5%, 1% and 2%) supported on alumina-silica mixed oxides with various molar ratios (Si/Al = 2.5; 5; 7; 10; 15). These catalysts will be characterized by numerous technics such as N₂ adsorption/desorption, Pyridine adsorption (acidity measure), FTIR, X-rays diffraction, AAS, TEM and SEM.Keywords: furfural, ruthenium, silica-alumina, biomass, biofuel
Procedia PDF Downloads 894370 The Effect of the Spinacia oleracea Extract on the Control of the Green Mold 'Penilillium digitatum' at the Post Harvested Citrus
Authors: Asma Chbani, Douaa Salim, Josephine Al Alam, Pascale De Caro
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Penicillium digitatum, the causal agent of citrus green mold, is responsible for 90% of post-harvest losses. Chemical fungicides remain the most used products for protection against this pathogen but are also responsible for damage to human health and the environment. The aim of this study is to evaluate the ability of Spinacia oleracea extract to serve as biological control agents, an alternative to harmful synthetic fungicides, against orange decay for storing fruit caused by P. digitatum. In this study, we studied the implication of a crude extract of a green plant, Spinacia oleracea, in the protection of oranges against P. digitatum. Thus, in vivo antifungal tests as well as adhesion test were done. For in vivo antifungal test, oranges were pulverized with the prepared crude extracts at different concentrations ranged from 25 g L⁻¹ to 200 g L⁻¹, contaminated by the fungus and then observed during 8 weeks for their macroscopic changes at 24°C. For adhesion test, the adhesion index is defined as the number of Penicillium digitatum spores fixed per orange cell. An index greater than 25 is the indicator of a strong adhesion, whereas for an index less than 10, the adhesion is low. Ten orange cells were examined in triplicate for each extract, and the averages of adherent cells were calculated. Obtained results showed an inhibitory activity of the Penicillium development with the aqueous extract of dry Spinacia oleracea with a concentration of 50 g L⁻¹ considered as the minimal protective concentration. The prepared extracts showed a greater inhibition of the development of P. digitatum up to 10 weeks, even greater than the fungicide control Nystatin. Adhesion test’s results showed that the adhesion of P. digitatum spores to the epidermal cells of oranges in the presence of the crude spinach leaves extract is weak; the mean of the obtained adhesion index was estimated to 2.7. However, a high adhesion was observed with water used a negative control. In conclusion, all these results confirm that the use of this green plant highly rich in chlorophyll having several phytotherapeutic activities, could be employed as a great treatment for protection of oranges against mold and also as an alternative for chemical fungicides.Keywords: Penicillium digitatum, Spinacia oleracea, oranges, biological control, postharvest diseases
Procedia PDF Downloads 1754369 Immunomodulation by Interleukin-10 Therapy in Mouse Airway Transplantation
Authors: Mohammaad Afzal Khan, Ghazi Abdulmalik Ashoor , Fatimah Alanazi, Talal Shamma, Abdullah Altuhami, Hala Abdalrahman Ahmed, Abdullah Mohammed Assiri, Dieter Clemens Broering
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Microvascular injuries during inflammation are key causes of transplant malfunctioning and permanent failure, which play a major role in the development of chronic rejection of the transplanted organ. Inflammation-induced microvascular loss is a promising area to investigate the decisive roles of regulatory and effector responses. The present study was designed to investigate the impact of IL-10 on immunotolerance, in particular, the microenvironment of the allograft during rejection. Here, we investigated the effects of IL-10 blockade/ reconstitution and serially monitored regulatory T cells (Tregs), graft microvasculature, and airway epithelium in rejecting airway transplants. We demonstrated that the blocking/reconstitution of IL-10 significantly modulates CD4+FOXP3+ Tregs, microvasculature, and airway epithelium during rejection. Our findings further highlighted that blockade of IL-10 upregulated proinflammatory cytokines, IL-2, IL-1β, IFN-γ, IL-15, and IL-23, but suppressed IL-5 secretion during rejection; however, reconstitution of IL-10 significantly upregulated CD4+FOXP3+ Tregs, tissue oxygenation/blood flow and airway repair. Collectively, these findings demonstrate a potential reparative modulation of IL-10 during microvascular and epithelial repair, which could provide a vital therapeutic window to rejecting transplants in clinical practice.Keywords: interleukin -10, regulatory T cells, allograft rejection, immunotolerance
Procedia PDF Downloads 1814368 A Cohesive Zone Model with Parameters Determined by Uniaxial Stress-Strain Curve
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A key issue of cohesive zone models is how to determine the cohesive zone model parameters based on real material test data. In this paper, uniaxial nominal stress-strain curve (SS curve) is used to determine two key parameters of a cohesive zone model (CZM): The maximum traction and the area under the curve of traction-separation law (TSL). To this end, the true SS curve is obtained based on the nominal SS curve, and the relationship between the nominal SS curve and TSL is derived based on an assumption that the stress for cracking should be the same in both CZM and the real material. In particular, the true SS curve after necking is derived from the nominal SS curve by taking the average of the power law extrapolation and the linear extrapolation, and a damage factor is introduced to offset the true stress reduction caused by the voids generated at the necking zone. The maximum traction of the TSL is equal to the maximum true stress calculated based on the damage factor at the end of hardening. In addition, a simple specimen is modeled by Abaqus/Standard to calculate the critical J-integral, and the fracture energy calculated by the critical J-integral represents the stored strain energy in the necking zone calculated by the true SS curve. Finally, the CZM parameters obtained by the present method are compared to those used in a previous related work for a simulation of the drop-weight tear test.Keywords: dynamic fracture, cohesive zone model, traction-separation law, stress-strain curve, J-integral
Procedia PDF Downloads 4764367 Increasing the Resilience of Cyber Physical Systems in Smart Grid Environments using Dynamic Cells
Authors: Andrea Tundis, Carlos García Cordero, Rolf Egert, Alfredo Garro, Max Mühlhäuser
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Resilience is an important system property that relies on the ability of a system to automatically recover from a degraded state so as to continue providing its services. Resilient systems have the means of detecting faults and failures with the added capability of automatically restoring their normal operations. Mastering resilience in the domain of Cyber-Physical Systems is challenging due to the interdependence of hybrid hardware and software components, along with physical limitations, laws, regulations and standards, among others. In order to overcome these challenges, this paper presents a modeling approach, based on the concept of Dynamic Cells, tailored to the management of Smart Grids. Additionally, a heuristic algorithm that works on top of the proposed modeling approach, to find resilient configurations, has been defined and implemented. More specifically, the model supports a flexible representation of Smart Grids and the algorithm is able to manage, at different abstraction levels, the resource consumption of individual grid elements on the presence of failures and faults. Finally, the proposal is evaluated in a test scenario where the effectiveness of such approach, when dealing with complex scenarios where adequate solutions are difficult to find, is shown.Keywords: cyber-physical systems, energy management, optimization, smart grids, self-healing, resilience, security
Procedia PDF Downloads 3314366 The Need For Higher Education Stem Integrated into the Social Science
Authors: Luis Fernando Calvo Prieto, Raul Herrero Martínez, Mónica Santamarta Llorente, Sergio Paniagua Bermejo
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The project that is presented starts from the questioning about the compartmentalization of knowledge that occurs in university higher education. There are several authors who describe the problems associated with this reality (Rodamillans, M) indicating a lack of integration of the knowledge acquired by students throughout the subjects taken in their university degree. Furthermore, this disintegration is accentuated by the enrollment system of some Faculties and/or Schools of Engineering, which allows the student to take subjects outside the recommended curricular path. This problem is accentuated in an ostentatious way when trying to find an integration between humanistic subjects and the world of experimental sciences or engineering. This abrupt separation between humanities and sciences can be observed in any study plan of Spanish degrees. Except for subjects such as economics or English, in the Faculties of Sciences and the Schools of Engineering, the absence of any humanistic content is striking. At some point it was decided that the only value to take into account when designing their study plans was “usefulness”, considering the humanities systematically useless for their training, and therefore banishing them from the study plans. forgetting the role they have on the capacity of both Leadership and Civic Humanism in our professionals of tomorrow. The teaching guides for the different subjects in the branch of science or engineering do not include any competency, not even transversal, related to leadership capacity or the need, in today's world, for social, civic and humanitarian knowledge part of the people who will offer medical, pharmaceutical, environmental, biotechnological or engineering solutions to a society that is generated thanks to more or less complex relationships based on human relationships and historical events that have occurred so far. If we want professionals who know how to deal effectively and rationally with their leadership tasks and who, in addition, find and develop an ethically civic sense and a humanistic profile in their functions and scientific tasks, we must not leave aside the importance that it has, for the themselves, know the causes, facts and consequences of key events in the history of humanity. The words of the humanist Paul Preston are well known: “he who does not know his history is condemned to repeat the mistakes of the past.” The idea, therefore, that today there can be men of science in the way that the scientists of the Renaissance were, becomes, at the very least, difficult to conceive. To think that a Leonardo da Vinci can be repeated in current times is a more than crazy idea; and although at first it may seem that the specialization of a professional is inevitable but beneficial, there are authors who consider (Sánchez Inarejos) that it has an extremely serious negative side effect: the entrenchment behind the different postulates of each area of knowledge, disdaining everything. what is foreign to it.Keywords: STEM, higher education, social sciences, history
Procedia PDF Downloads 694365 Modelling and Control of Binary Distillation Column
Authors: Narava Manose
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Distillation is a very old separation technology for separating liquid mixtures that can be traced back to the chemists in Alexandria in the first century A. D. Today distillation is the most important industrial separation technology. By the eleventh century, distillation was being used in Italy to produce alcoholic beverages. At that time, distillation was probably a batch process based on the use of just a single stage, the boiler. The word distillation is derived from the Latin word destillare, which means dripping or trickling down. By at least the sixteenth century, it was known that the extent of separation could be improved by providing multiple vapor-liquid contacts (stages) in a so called Rectifactorium. The term rectification is derived from the Latin words rectefacere, meaning to improve. Modern distillation derives its ability to produce almost pure products from the use of multi-stage contacting. Throughout the twentieth century, multistage distillation was by far the most widely used industrial method for separating liquid mixtures of chemical components.The basic principle behind this technique relies on the different boiling temperatures for the various components of the mixture, allowing the separation between the vapor from the most volatile component and the liquid of other(s) component(s). •Developed a simple non-linear model of a binary distillation column using Skogestad equations in Simulink. •We have computed the steady-state operating point around which to base our analysis and controller design. However, the model contains two integrators because the condenser and reboiler levels are not controlled. One particular way of stabilizing the column is the LV-configuration where we use D to control M_D, and B to control M_B; such a model is given in cola_lv.m where we have used two P-controllers with gains equal to 10.Keywords: modelling, distillation column, control, binary distillation
Procedia PDF Downloads 2824364 Significance of Molecular Autophagic Pathway in Gaucher Disease Pathology
Authors: Ozlem Oral, Emre Taskin, Aysel Yuce, Serap Dokmeci, Devrim Gozuacik
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Autophagy is an evolutionary conserved lysosome-dependent catabolic pathway, responsible for the degradation of long-lived proteins, abnormal aggregates and damaged organelles which cannot be degraded by the ubiquitin-proteasome system. Lysosomes degrade the substrates through the activity of lysosomal hydrolases and lysosomal membrane-bound proteins. Mutations in the coding region of these proteins cause malfunctional lysosomes, which contributes to the pathogenesis of lysosomal storage diseases. Gaucher disease is a lysosomal storage disease resulting from the mutation of a lysosomal membrane-associated glycoprotein called glucocerebrosidase and its cofactor saposin C. The disease leads to intracellular accumulation of glucosylceramide and other glycolipids. Because of the essential role of lysosomes in autophagic degradation, Gaucher disease may directly be linked to this pathway. In this study, we investigated the expression of autophagy and/or lysosome-related genes and proteins in fibroblast cells isolated from patients with different mutations. We carried out confocal microscopy analysis and examined autophagic flux by utilizing the differential pH sensitivities of RFP and GFP in mRFP-GFP-LC3 probe. We also evaluated lysosomal pH by active lysosome staining and lysosomal enzyme activity. Beside lysosomes, we also performed proteasomal activity and cell death analysis in patient samples. Our data showed significant attenuation in the expression of key autophagy-related genes and accumulation of their proteins in mutant cells. We found decreased the ability of autophagosomes to fuse with lysosomes, associated with elevated lysosomal pH and reduced lysosomal enzyme activity. Proteasomal degradation and cell death analysis showed reduced proteolytic activity of the proteasome, which consequently leads to increased susceptibility to cell death. Our data indicate that the major degradation pathways are affected by multifunctional lysosomes in mutant patient cells and may underlie in the mechanism of clinical severity of Gaucher patients. (This project is supported by TUBITAK-3501-National Young Researchers Career Development Program, Project No: 112T130).Keywords: autophagy, Gaucher's disease, glucocerebrosidase, mutant fibroblasts
Procedia PDF Downloads 3274363 The Endocrinology of Obesity and Dejenerative Joint Disease
Authors: Kebret Kebede, Anthony Scinta
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Obesity is the most prevalent global problem that continues to rise at alarming rates both in the industrialized and developing countries. Adipose tissue is an endocrine tissue that secretes numerous chemical signals, hormones, lipids, cytokines and coagulation factors as well as prompting insulin resistance which is a primary contributor to Type II Diabetes- one of its most common adverse effects on health. Other hormones whose levels are linked to obesity and nutritional state are leptin, IGF-1, and adiponectin. Several studies indicate that obesity is the leading cause of high levels of cholesterol that leads to fatty liver disease, gallstones, hypertension, increased risk for cancer and degenerative joint disease that primarily affects the weight bearing joints of the lower extremities. The activation of inflammatory pathways promotes synovial pathology that results in accelerated degeneration of the joints. The study examines the prevalence of obesity in the US female population in comparison to that of the developing world and its emergence as a significant and potentially modifiable risk factor in degenerative disease of the hip and knee joints that has resulted in staggering healthcare cost. Studies have shown that as the prevalence of obesity rises, we continue to see a rise in degenerative joint disease. The percentage of arthritis cases linked directly to obesity has risen from 3 percent in 1971 to 18 percent in 2002. A person with obesity is around 60 percent more likely to develop arthritis than someone of normal body weight. In women, obesity is associated with increased mortality from breast, cervical, endometrial and ovarian cancer that may accompany debilitating joint diseases and restricted mobility.Keywords: obesity, endocrine, degenerative, mortality, joint diseases, cancer, debilitating, mobility
Procedia PDF Downloads 4534362 Determination of Cohesive Zone Model’s Parameters Based On the Uniaxial Stress-Strain Curve
Authors: Y. J. Wang, C. Q. Ru
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A key issue of cohesive zone models is how to determine the cohesive zone model (CZM) parameters based on real material test data. In this paper, uniaxial nominal stress-strain curve (SS curve) is used to determine two key parameters of a cohesive zone model: the maximum traction and the area under the curve of traction-separation law (TSL). To this end, the true SS curve is obtained based on the nominal SS curve, and the relationship between the nominal SS curve and TSL is derived based on an assumption that the stress for cracking should be the same in both CZM and the real material. In particular, the true SS curve after necking is derived from the nominal SS curve by taking the average of the power law extrapolation and the linear extrapolation, and a damage factor is introduced to offset the true stress reduction caused by the voids generated at the necking zone. The maximum traction of the TSL is equal to the maximum true stress calculated based on the damage factor at the end of hardening. In addition, a simple specimen is simulated by Abaqus/Standard to calculate the critical J-integral, and the fracture energy calculated by the critical J-integral represents the stored strain energy in the necking zone calculated by the true SS curve. Finally, the CZM parameters obtained by the present method are compared to those used in a previous related work for a simulation of the drop-weight tear test.Keywords: dynamic fracture, cohesive zone model, traction-separation law, stress-strain curve, J-integral
Procedia PDF Downloads 5164361 Effect of Velocity Slip on Two Phase Flow in an Eccentric Annular Region
Authors: Umadevi B., Dinesh P. A., Indira. R., Vinay C. V.
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A mathematical model is developed to study the simultaneous effects of particle drag and slip parameter on the velocity as well as rate of flow in an annular cross sectional region bounded by two eccentric cylinders. In physiological flows this phenomena can be observed in an eccentric catheterized artery with inner cylinder wall is impermeable and outer cylinder wall is permeable. Blood is a heterogeneous fluid having liquid phase consisting of plasma in which a solid phase of suspended cells and proteins. Arterial wall gets damaged due to aging and lipid molecules get deposited between damaged tissue cells. Blood flow increases towards the damaged tissues in the artery. In this investigation blood is modeled as two phase fluid as one is a fluid phase and the other is particulate phase. The velocity of the fluid phase and rate of flow are obtained by transforming eccentric annulus to concentric annulus with the conformal mapping. The formulated governing equations are analytically solved for the velocity and rate of flow. The numerical investigations are carried out by varying eccentricity parameter, slip parameter and drag parameter. Enhancement of slip parameter signifies loss of fluid then the velocity and rate of flow will be decreased. As particulate drag parameter increases then the velocity as well as rate flow decreases. Eccentricity facilitates transport of more fluid then the velocity and rate of flow increases.Keywords: catheter, slip parameter, drag parameter, eccentricity
Procedia PDF Downloads 5274360 Cytotoxic Activity of Extracts from Hibiscus sabdariffa Leaves against Women’s Cancer Cell Lines
Authors: Patsorn Worawattananutai, Srisopa Ruangnoo, Arunporn Itharat
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Hibiscus sabdariffa (HS) leaves are vegetables which are extensively used as blood tonic and laxatives in Thai traditional medicine. They are popularly used as healthy sour soup for prevention of chronic diseases such as cancer. Therefore, the cytotoxic activity of different extracts of fresh and dried Hibiscus sabdariffa leaves were investigated via the sulforhodamine B (SRB) assay against three types of women’s cancer cell lines, namely the human cervical adenocarcinoma cell line (HeLa), the human ovarian adenocarcinoma cell line (SKOV-3), and the human breast adenocarcinoma cell line (MCF-7). Extraction methods were squeezing, boiling with water and maceration with 95% or 50% ethanol. The 95% ethanolic extracts of Hibiscus sabdariffa dry leaves (HSDE95) showed the highest cytotoxicity against all types of women’s cancer cell lines with the IC50 values in range 7.51±0.33 to 12.13±1.85 µg/ml. Its IC50 values against SKOV-3, HeLa and MCF-7 were 7.51±0.33, 9.44±1.41 and 12.13±1.85 µg/ml, respectively. In these results, this extract can be classified as “active” according to the NCI guideline which indicated that IC50 values of the active cytotoxic plant extracts have to be beneath 20 µg/ml. Thus, HSDE95 was concluded to be a potent cytotoxic drug for all women’s cancer cells. This extract should be further investigated to isolate active compounds against women’s cancer cells.Keywords: breast adenocarcinoma, cervical adenocarcinoma, cytotoxic activity, Hibiscus sabdariffa, ovarian adenocarcinoma
Procedia PDF Downloads 6044359 In vitro Evaluation of the Synergistic Antiviral Activity of Amantadine Coupled with Magnesium Lithospermate B against Enterovirus 71 Infection
Authors: Wen-Yu Lin, Yi-Ching Chung, Jhao-Ren Lin, Tzyy-Rong Jinn
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It is well known that enterovirus 71(EV71) causes recurring outbreaks of hand, foot and mouth disease and encephalitis leading to complications or death in young children. And, several enterovirus 71 (EV71) of hand foot and mouth disease (HFMD) with high mortalities occurred in Asia country, such as Hong Kung (1985), Malaysia (1997), Taiwan (1998) and China (2008) that EV71 results in severe neurological complications and sudden death in infants and young children. However, there are still no effective drugs and vaccines to reduce and inhibit EV71 infection. Therefore, the development of specific and effective antiviral strategies against EV71 has become an urgent issue for the protection of children from the hazards of the HFMD. As reported, amantadine is effective in prophylaxis and treatment of the EV71 infections. Thus, the aim of this study was to further evaluate the synergistic antiviral activity of amantadine coupled with magnesium lithospermate B (MLB) against enterovirus 71 infection. In a preliminary test, it is shown that the infected RD cells were treated with amantadine after virus absorption, at concentrations of 3 and 5µM of amantadine suppressed EV71-induced CPE to 13% and 23%, respectively at MOI of 3. Alternatively, at concentrations of 5µg/ml of MLB combined with 3 and 5 µM of amantadine apparently suppressed EV71-induced CPE to 45% and 63%, respectively at MOI of 3. Thus, amantadine coupled with MLB may have the potential for further study to development as the chemopreventive reagents against EV71 infection.Keywords: amantadine, Enterovirus 71, magnesium lithospermate B, RD cells, synergistic effects
Procedia PDF Downloads 2474358 Cardiorespiratory Fitness and the Cardiometabolic Profile in Inactive Obese Postmenopausal Women: A MONET Study
Authors: Ahmed Ghachem, Johann Colomba, Denis Prud'homme, Martin Brochu
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Background: Inactive obese postmenopausal women, are at greater risk for metabolic complications. On the other hand, high levels of cardiorespiratory fitness (CRF) are associated with a lower risk of metabolic complications. Objective: To compare inactive obese postmenopausal women displaying ‘lower’ vs ‘higher’ levels of CRF for body composition, metabolic profile, inflammatory profile and measures of energy expenditure. Methods: 132 women (age: 57.6 ± 4.8 yrs; BMI: 32.3 ± 4.6 kg/m2; Peak VO2: 17.81 ± 3.02 ml O2•kg-1•min-1) were studied. They were first divided into tertiles based on their CRF. Then, women in the first (< 16.51 ml O2•min-1•kg-1) and second tertiles (16.51 to 19.22 ml O2•min-1•kg-1) were combined (N= 88), and compared with those in the third tertile (> 19.22 ml O2•min-1•kg-1) (N= 44). Variables of interest were: Peak VO2 (stationary bike), body composition (DXA), body fat distribution (CT scan), glucose homeostasis (fasting state and euglycemic/ hyperinsulinemic clamp), fasting lipids, resting blood pressure, inflammatory profile and energy expenditure (DLW). Results: Both CRF groups (lower= 16.0 ± 2.0 ml O2•kg-1•min-1 vs higher= 21.2 ± 1.7 ml O2•kg-1•min-1; p < 0.001) were similar for age. Significant differences were observed between groups for body composition; with lower values for body weight, BMI, fat mass and visceral fat in women with higher CRF (p between 0.001 and 0.005). Also, women with higher CRF had lower values for fasting insulin (13.4 ± 4.5 vs 15.6 ± 6.6 μU/ml; p = 0.03) and CRP levels (2.31 ± 1.97 vs 3.83 ± 3.24 mg/liter; p = 0.001); and higher values for glucose disposal (6.71 ± 1.78 vs 5.92 ± 1.67 mg/kg/min; p = 0.01). However, these differences were no longer significant after controlling for visceral adipose tissue accumulations. Finally, no significant difference was observed between groups for the other variables of interest. Conclusion: Our results suggest that, among inactive overweight/obese postmenopausal women, those with higher CRF levels have a better metabolic profile; which is caused by lower visceral fat accumulations.Keywords: cardiorespiratory fitness, metabolic profile, menopause, obesity
Procedia PDF Downloads 2704357 Effects of a Bacteria-Based Probiotic on Subpopulations of Peripheral Leukocytes and Their Interleukin mRNA Expression in Calves
Authors: Abdul Qadir Qadis, Satoru Goya, Minoru Yatsu, Yu-uki Yoshida, Toshihiro Ichijo, Shigeru Sato
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Bacterial probiotics are known to modulate the gut-associated lymphoid and epithelial tissue response to enhance the activities of intestinal and systemic immune system in human and animals. In cattle, the immune-stimulatory effects of probiotics have been evaluated during intestinal disorders. To investigate the effects of probiotic on the function of peripheral blood mononuclear cells, eight healthy Holstein calves (10 ± 3 weeks) were assigned to a 4 × 2 experimental design. The probiotic, consisting of Lactobacillus plantarum, Enterococcus faecium and Clostridium butyricum, was administered orally at 3.0 g/100 kg body weight to calves once daily for 5 consecutive days. Calves given no probiotic served as the control. In the treatment group, increases in numbers of CD282+ monocytes, CD3+ T-cells and CD4+, CD8+ and WC1+ γδ T- cell subsets were noted on day 7 post-placement compared to pre-dose day and the control group. Expression of interleukin-6, interferon-gamma and tumor necrosis factor-alpha was elevated in peripheral leukocytes on days 7 and 14. These results suggest that peripheral blood leukocytes in healthy calves may be stimulated via the gastrointestinal microbiota, which was increased by the oral probiotic treatment. The 5-day repeated administration of a bacterial probiotic may enhance cellular immune function in weaned calves.Keywords: bacterial-probiotic, calf, interleukin, leukocyte
Procedia PDF Downloads 6634356 Novel Phenolic Biopolyether with Potential Therapeutic Effect
Authors: V.Barbakadze, L.Gogilashvili, L.Amiranashvili, M.Merlani, K.Mulkijanyan
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The high-molecular fractions from the several species of two genera (Symphytum and Anchusa) of Boraginaceae family Symphytum asperum, S. caucasicum, S. officinale, and Anchusa italica were isolated. According to IR, 13C and 1H NMR, 2D heteronuclear 1H/13C HSQC spectral data and 1D NOE experiment, the main structural element of these preparations was found to be a regularly substituted polyoxyethylene, namely poly[3-(3,4-dihydroxyenyl)glyceric acid] (PDPGA) or poly[oxy-1-carboxy-2-(3,4-dihydroxyphenyl)ethylene]. Such caffeic acid-derived biopolymer to our knowledge has not been known and has been identified for the first time. This compound represents a new class of natural polyethers with a residue of 3-(3,4-dihydroxyphenyl)glyceric acid as the repeating unit. Most of the carboxylic groups of PDPGA from A. italica unlike the polymer of S. asperum, S. caucasicum, and S. officinale are methylated. The 2D DOSY experiment gave the similar diffusion coefficient for the methylated and non-methylated signals of A. italica PDPGA. Both sets of signals fell in the same horizontal. This would imply a similar molecular weight for methylated and non-methylated polymers. This was further evidenced by graphic representations of the intensity decay of the 1H signals of aromatic H-2″ and H-1 at δ 7.16 and 5.24 and that of the methoxy group at δ 3.85. These three signals essentially showed the same curve shape. According to results of in vitro and in vivo experiments PDPGA of S.asperum and S.caucasicum could be considered as potential anti-inflammatory, wound healing and anti-cancer therapeutic agent.Keywords: caffeic acid-derived polyether, poly[3-(3, 4-dihydroxyphenyl)glyceric acid], poly[oxy-1-carboxy-2-(3, 4-dihydroxyphenyl)ethylene], symphytum, anchusa
Procedia PDF Downloads 4104355 Characterizing and Developing the Clinical Grade Microbiome Assay with a Robust Bioinformatics Pipeline for Supporting Precision Medicine Driven Clinical Development
Authors: Danyi Wang, Andrew Schriefer, Dennis O'Rourke, Brajendra Kumar, Yang Liu, Fei Zhong, Juergen Scheuenpflug, Zheng Feng
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Purpose: It has been recognized that the microbiome plays critical roles in disease pathogenesis, including cancer, autoimmune disease, and multiple sclerosis. To develop a clinical-grade assay for exploring microbiome-derived clinical biomarkers across disease areas, a two-phase approach is implemented. 1) Identification of the optimal sample preparation reagents using pre-mixed bacteria and healthy donor stool samples coupled with proprietary Sigma-Aldrich® bioinformatics solution. 2) Exploratory analysis of patient samples for enabling precision medicine. Study Procedure: In phase 1 study, we first compared the 16S sequencing results of two ATCC® microbiome standards (MSA 2002 and MSA 2003) across five different extraction kits (Kit A, B, C, D & E). Both microbiome standards samples were extracted in triplicate across all extraction kits. Following isolation, DNA quantity was determined by Qubit assay. DNA quality was assessed to determine purity and to confirm extracted DNA is of high molecular weight. Bacterial 16S ribosomal ribonucleic acid (rRNA) amplicons were generated via amplification of the V3/V4 hypervariable region of the 16S rRNA. Sequencing was performed using a 2x300 bp paired-end configuration on the Illumina MiSeq. Fastq files were analyzed using the Sigma-Aldrich® Microbiome Platform. The Microbiome Platform is a cloud-based service that offers best-in-class 16S-seq and WGS analysis pipelines and databases. The Platform and its methods have been extensively benchmarked using microbiome standards generated internally by MilliporeSigma and other external providers. Data Summary: The DNA yield using the extraction kit D and E is below the limit of detection (100 pg/µl) of Qubit assay as both extraction kits are intended for samples with low bacterial counts. The pre-mixed bacterial pellets at high concentrations with an input of 2 x106 cells for MSA-2002 and 1 x106 cells from MSA-2003 were not compatible with the kits. Among the remaining 3 extraction kits, kit A produced the greatest yield whereas kit B provided the least yield (Kit-A/MSA-2002: 174.25 ± 34.98; Kit-A/MSA-2003: 179.89 ± 30.18; Kit-B/MSA-2002: 27.86 ± 9.35; Kit-B/MSA-2003: 23.14 ± 6.39; Kit-C/MSA-2002: 55.19 ± 10.18; Kit-C/MSA-2003: 35.80 ± 11.41 (Mean ± SD)). Also, kit A produced the greatest yield, whereas kit B provided the least yield. The PCoA 3D visualization of the Weighted Unifrac beta diversity shows that kits A and C cluster closely together while kit B appears as an outlier. The kit A sequencing samples cluster more closely together than both the other kits. The taxonomic profiles of kit B have lower recall when compared to the known mixture profiles indicating that kit B was inefficient at detecting some of the bacteria. Conclusion: Our data demonstrated that the DNA extraction method impacts DNA concentration, purity, and microbial communities detected by next-generation sequencing analysis. Further microbiome analysis performance comparison of using healthy stool samples is underway; also, colorectal cancer patients' samples will be acquired for further explore the clinical utilities. Collectively, our comprehensive qualification approach, including the evaluation of optimal DNA extraction conditions, the inclusion of positive controls, and the implementation of a robust qualified bioinformatics pipeline, assures accurate characterization of the microbiota in a complex matrix for deciphering the deep biology and enabling precision medicine.Keywords: 16S rRNA sequencing, analytical validation, bioinformatics pipeline, metagenomics
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