Search results for: molecular diversity
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3648

Search results for: molecular diversity

438 Cytokine Profiling in Cultured Endometrial Cells after Hormonal Treatment

Authors: Mark Gavriel, Ariel J. Jaffa, Dan Grisaru, David Elad

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The human endometrium-myometrium interface (EMI) is the uterine inner barrier without a separatig layer. It is composed of endometrial epithelial cells (EEC) and endometrial stromal cells (ESC) in the endometrium and myometrial smooth muscle cells (MSMC) in the myometrium. The EMI undergoes structural remodeling during the menstruation cycle which are essential for human reproduction. Recently, we co-cultured a layer-by-layer in vitro model of EEC, ESC and MSMC on a synthetic membrane for mechanobiology experiments. We also treated the model with progesterone and β-estradiol in order to mimic the in vivo receptive uterus In the present study we analyzed the cytokines profile in a single layer of EEC the hormonal treated in vitro model of the EMI. The methodologies of this research include simple tissue-engineering . First, we cultured commercial EEC (RL95-2, ATCC® CRL-1671™) in 24-wellplate. Then, we applied an hormonal stimuli protocol with 17-β-estradiol and progesterone in time dependent concentration according to the human physiology that mimics the menstrual cycle. We collected cell supernatant samples of control, pre-ovulation, ovulation and post-ovulaton periods for analysis of the secreted proteins and cytokines. The cytokine profiling was performed using the Proteome Profiler Human XL Cytokine Array Kit (R&D Systems, Inc., USA) that can detect105 human soluble cytokines. The relative quantification of all the cytokines will be analyzed using xMAP – LUMINEX. We conducted a fishing expedition with the 4 membranes Proteome Profiler. We processed the images, quantified the spots intensity and normalized these values by the negative control and reference spots at the membrane. Analyses of the relative quantities that reflected change higher than 5% of the control points of the kit revealed the The results clearly showed that there are significant changes in the cytokine level for inflammation and angiogenesis pathways. Analysis of tissue-engineered models of the uterine wall will enable deeper investigation of molecular and biomechanical aspects of early reproductive stages (e.g. the window of implantation) or developments of pathologies.

Keywords: tissue-engineering, hormonal stimuli, reproduction, multi-layer uterine model, progesterone, β-estradiol, receptive uterine model, fertility

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437 Changes in Geospatial Structure of Households in the Czech Republic: Findings from Population and Housing Census

Authors: Jaroslav Kraus

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Spatial information about demographic processes are a standard part of outputs in the Czech Republic. That was also the case of Population and Housing Census which was held on 2011. This is a starting point for a follow up study devoted to two basic types of households: single person households and households of one completed family. Single person households and one family households create more than 80 percent of all households, but the share and spatial structure is in long-term changing. The increase of single households is results of long-term fertility decrease and divorce increase, but also possibility of separate living. There are regions in the Czech Republic with traditional demographic behavior, and regions like capital Prague and some others with changing pattern. Population census is based - according to international standards - on the concept of currently living population. Three types of geospatial approaches will be used for analysis: (i) firstly measures of geographic distribution, (ii) secondly mapping clusters to identify the locations of statistically significant hot spots, cold spots, spatial outliers, and similar features and (iii) finally analyzing pattern approach as a starting point for more in-depth analyses (geospatial regression) in the future will be also applied. For analysis of this type of data, number of households by types should be distinct objects. All events in a meaningful delimited study region (e.g. municipalities) will be included in an analysis. Commonly produced measures of central tendency and spread will include: identification of the location of the center of the point set (by NUTS3 level); identification of the median center and standard distance, weighted standard distance and standard deviational ellipses will be also used. Identifying that clustering exists in census households datasets does not provide a detailed picture of the nature and pattern of clustering but will be helpful to apply simple hot-spot (and cold spot) identification techniques to such datasets. Once the spatial structure of households will be determined, any particular measure of autocorrelation can be constructed by defining a way of measuring the difference between location attribute values. The most widely used measure is Moran’s I that will be applied to municipal units where numerical ratio is calculated. Local statistics arise naturally out of any of the methods for measuring spatial autocorrelation and will be applied to development of localized variants of almost any standard summary statistic. Local Moran’s I will give an indication of household data homogeneity and diversity on a municipal level.

Keywords: census, geo-demography, households, the Czech Republic

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436 Study on the Rapid Start-up and Functional Microorganisms of the Coupled Process of Short-range Nitrification and Anammox in Landfill Leachate Treatment

Authors: Lina Wu

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The excessive discharge of nitrogen in sewage greatly intensifies the eutrophication of water bodies and poses a threat to water quality. Nitrogen pollution control has become a global concern. Currently, the problem of water pollution in China is still not optimistic. As a typical high ammonia nitrogen organic wastewater, landfill leachate is more difficult to treat than domestic sewage because of its complex water quality, high toxicity, and high concentration.Many studies have shown that the autotrophic anammox bacteria in nature can combine nitrous and ammonia nitrogen without carbon source through functional genes to achieve total nitrogen removal, which is very suitable for the removal of nitrogen from leachate. In addition, the process also saves a lot of aeration energy consumption than the traditional nitrogen removal process. Therefore, anammox plays an important role in nitrogen conversion and energy saving. The process composed of short-range nitrification and denitrification coupled an ammo ensures the removal of total nitrogen and improves the removal efficiency, meeting the needs of the society for an ecologically friendly and cost-effective nutrient removal treatment technology. Continuous flow process for treating late leachate [an up-flow anaerobic sludge blanket reactor (UASB), anoxic/oxic (A/O)–anaerobic ammonia oxidation reactor (ANAOR or anammox reactor)] has been developed to achieve autotrophic deep nitrogen removal. In this process, the optimal process parameters such as hydraulic retention time and nitrification flow rate have been obtained, and have been applied to the rapid start-up and stable operation of the process system and high removal efficiency. Besides, finding the characteristics of microbial community during the start-up of anammox process system and analyzing its microbial ecological mechanism provide a basis for the enrichment of anammox microbial community under high environmental stress. One research developed partial nitrification-Anammox (PN/A) using an internal circulation (IC) system and a biological aerated filter (BAF) biofilm reactor (IBBR), where the amount of water treated is closer to that of landfill leachate. However, new high-throughput sequencing technology is still required to be utilized to analyze the changes of microbial diversity of this system, related functional genera and functional genes under optimal conditions, providing theoretical and further practical basis for the engineering application of novel anammox system in biogas slurry treatment and resource utilization.

Keywords: nutrient removal and recovery, leachate, anammox, partial nitrification

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435 “Student Veterans’ Transition to Nursing Education: Barriers and Facilitators

Authors: Bruce Hunter

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Background: The transition for student veterans from military service to higher education can be a challenging endeavor, especially for those pursuing an education in nursing. While the experiences and perspectives of each student veteran is unique, their successful integration into an academic environment can be influenced by a complex array of barriers and facilitators. This mixed-methods study aims to explore the themes and concepts that can be found in the transition experiences of student veterans in nursing education, with a focus on identifying the barriers they face and the facilitators that support their success. Methods: This study utilizes an explanatory mixed-methods approach. The research participants include student veterans enrolled in nursing programs across three academic institutions in the Southeastern United States. Quantitative Phase: A Likert scale instrument is distributed to a sample of student veterans in nursing programs. The survey assesses demographic information, academic experiences, social experiences, and perceptions of institutional support. Quantitative data is analyzed using descriptive statistics to assess demographics and to identify barriers and facilitators to the transition. Qualitative Phase: Two open-ended questions were posed to student veterans to explore their lived experiences, barriers, and facilitators during the transition to nursing education and to further explain the quantitative findings. Thematic analysis with line-by-line coding is employed to identify recurring themes and narratives that may shed light on the barriers and facilitators encountered. Results: This study found that the successful academic integration of student veterans lies in recognizing the diversity of values and attitudes among student veterans, understanding the potential challenges they face, and engaging in initiative-taking steps to create an inclusive and supportive academic environment that accommodates the unique experiences of this demographic. Addressing these academic and social integration concerns can contribute to a more understanding environment for student veterans in the BSN program. Conclusion: Providing support during this transitional period is crucial not only for retaining veterans, but also for bolstering their success in achieving the status of registered nurses. Acquiring an understanding of military culture emerges as an essential initial step for nursing faculty in student veteran retention and for successful completion of their programs. Participants found that their transition experience lacked meaningful social interactions, which could foster a positive learning environment, enhance their emotional well-being, and could contribute significantly to their overall success and satisfaction in their nursing education journey. Recognizing and promoting academic and social integration is important in helping veterans experience a smooth transition into and through the unfamiliar academic environment of nursing education.

Keywords: nursing, education, student veterans, barriers, facilitators

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434 What Are the Problems in the Case of Analysis of Selenium by Inductively Coupled Plasma Mass Spectrometry in Food and Food Raw Materials?

Authors: Béla Kovács, Éva Bódi, Farzaneh Garousi, Szilvia Várallyay, Dávid Andrási

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For analysis of elements in different food, feed and food raw material samples generally a flame atomic absorption spectrometer (FAAS), a graphite furnace atomic absorption spectrometer (GF-AAS), an inductively coupled plasma optical emission spectrometer (ICP-OES) and an inductively coupled plasma mass spectrometer (ICP-MS) are applied. All the analytical instruments have different physical and chemical interfering effects analysing food and food raw material samples. The smaller the concentration of an analyte and the larger the concentration of the matrix the larger the interfering effects. Nowadays, it is very important to analyse growingly smaller concentrations of elements. From the above analytical instruments generally the inductively coupled plasma mass spectrometer is capable of analysing the smallest concentration of elements. The applied ICP-MS instrument has Collision Cell Technology (CCT) also. Using CCT mode certain elements have better detection limits with 1-3 magnitudes comparing to a normal ICP-MS analytical method. The CCT mode has better detection limits mainly for analysis of selenium (arsenic, germanium, vanadium, and chromium). To elaborate an analytical method for selenium with an inductively coupled plasma mass spectrometer the most important interfering effects (problems) were evaluated: 1) isobaric elemental, 2) isobaric molecular, and 3) physical interferences. Analysing food and food raw material samples an other (new) interfering effect emerged in ICP-MS, namely the effect of various matrixes having different evaporation and nebulization effectiveness, moreover having different quantity of carbon content of food, feed and food raw material samples. In our research work the effect of different water-soluble compounds furthermore the effect of various quantity of carbon content (as sample matrix) were examined on changes of intensity of selenium. So finally we could find “opportunities” to decrease the error of selenium analysis. To analyse selenium in food, feed and food raw material samples, the most appropriate inductively coupled plasma mass spectrometer is a quadrupole instrument applying a collision cell technique (CCT). The extent of interfering effect of carbon content depends on the type of compounds. The carbon content significantly affects the measured concentration (intensities) of Se, which can be corrected using internal standard (arsenic or tellurium).

Keywords: selenium, ICP-MS, food, food raw material

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433 Effect of Chemical Fertilizer on Plant Growth-Promoting Rhizobacteria in Wheat

Authors: Tessa E. Reid, Vanessa N. Kavamura, Maider Abadie, Adriana Torres-Ballesteros, Mark Pawlett, Ian M. Clark, Jim Harris, Tim Mauchline

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The deleterious effect of chemical fertilizer on rhizobacterial diversity has been well documented using 16S rRNA gene amplicon sequencing and predictive metagenomics. Biofertilization is a cost-effective and sustainable alternative; improving strategies depends on isolating beneficial soil microorganisms. Although culturing is widespread in biofertilization, it is unknown whether the composition of cultured isolates closely mirrors native beneficial rhizobacterial populations. This study aimed to determine the relative abundance of culturable plant growth-promoting rhizobacteria (PGPR) isolates within total soil DNA and how potential PGPR populations respond to chemical fertilization in a commercial wheat variety. It was hypothesized that PGPR will be reduced in fertilized relative to unfertilized wheat. Triticum aestivum cv. Cadenza seeds were sown in a nutrient depleted agricultural soil in pots treated with and without nitrogen-phosphorous-potassium (NPK) fertilizer. Rhizosphere and rhizoplane samples were collected at flowering stage (10 weeks) and analyzed by culture-independent (amplicon sequence variance (ASV) analysis of total rhizobacterial DNA) and -dependent (isolation using growth media) techniques. Rhizosphere- and rhizoplane-derived microbiota culture collections were tested for plant growth-promoting traits using functional bioassays. In general, fertilizer addition decreased the proportion of nutrient-solubilizing bacteria (nitrate, phosphate, potassium, iron and, zinc) isolated from rhizocompartments in wheat, whereas salt tolerant bacteria were not affected. A PGPR database was created from isolate 16S rRNA gene sequences and searched against total soil DNA, revealing that 1.52% of total community ASVs were identified as culturable PGPR isolates. Bioassays identified a higher proportion of PGPR in non-fertilized samples (rhizosphere (49%) and rhizoplane (91%)) compared to fertilized samples (rhizosphere (21%) and rhizoplane (19%)) which constituted approximately 1.95% and 1.25% in non-fertilized and fertilized total community DNA, respectively. The analyses of 16S rRNA genes and deduced functional profiles provide an in-depth understanding of the responses of bacterial communities to fertilizer; this study suggests that rhizobacteria, which potentially benefit plants by mobilizing insoluble nutrients in soil, are reduced by chemical fertilizer addition. This knowledge will benefit the development of more targeted biofertilization strategies.

Keywords: bacteria, fertilizer, microbiome, rhizoplane, rhizosphere

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432 Effect of a Synthetic Platinum-Based Complex on Autophagy Induction in Leydig TM3 Cells

Authors: Ezzati Givi M., Hoveizi E., Nezhad Marani N.

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Platinum-based anticancer therapeutics are the most widely used drugs in clinical chemotherapy but have major limitations and various side effects in clinical applications. Gonadotoxicity and sterility is one of the most common complications for cancer survivors, which seem to be drug-specific and dose-related. Therefore, many efforts have been dedicated to discovering a new structure of platinum-based anticancer agents with improved therapeutic index, fewer side effects. In this regard, new Pt(II)-phosphane complexes containing heterocyclic thionate ligands (PCTL) have been synthesized, which show more potent antitumor activities in comparison to cisplatin. Cisplatin, the best leading metal-based antitumor drug in the field, induces testicular toxicity on Leydig and Sertoli cells leading to serious side effects such as azoospermia and infertility. Therefore in the present study, we aimed to investigate the cytotoxicity effect of PCTL on mice TM4 Sertoli cells with particular emphasis on the role of autophagy in comparison to cisplatin. In this study, an MTT assay was performed to evaluate the IC50 of PCTL and to analyze the TM3 Leydig cell's viability. Cells morphology was evaluated via invert microscope and Changing in morphology for nuclei swelling or autophagic vacuoles formation were assessed by DAPI and MDC staining. Testosterone production in the culture medium was measured using an ELISA kit. Finally, the expression of Autophagy-related genes, Atg5, Beclin1 and p62, were analyzed by qPCR. Based on the obtained results by MTT, the IC50 value of PCTL was 50 μM in TM3 cells and cytotoxic effects was in a dose- and time-dependent manner. Cells morphological changes investigated by inverted microscopy, DAPI, and MDC staining which showed the cytotoxic concentrations of PCTL was significantly higher than cisplatin in the treated TM3 Leydig cells. The results of PCR showed a lack of expression of the p62, Atg5 and Beclin1 gene in TM3 cells treated with PCTL in comparison to cisplatin and control groups. It should be noted that the effects of 25 μM PCTL concentration on TM3 cells have been associated with increased testosterone production and secretion, which requires further study to explain the possible causes and involved molecular mechanisms. The results of the study showed that the PCTL had less-lethal effects on TM3 cells in comparison to cisplatin and probably did not induce autophagy in TM3 cells.

Keywords: platinum-based anticancer agents, cisplatin, Leydig TM3 cells, autophagy

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431 BLS-2/BSL-3 Laboratory for Diagnosis of Pathogens on the Colombia-Ecuador Border Region: A Post-COVID Commitment to Public Health

Authors: Anderson Rocha-Buelvas, Jaqueline Mena Huertas, Edith Burbano Rosero, Arsenio Hidalgo Troya, Mauricio Casas Cruz

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COVID-19 is a disruptive pandemic for the public health and economic system of whole countries, including Colombia. Nariño Department is the southwest of the country and draws attention to being on the border with Ecuador, constantly facing demographic transition affecting infections between countries. In Nariño, the early routine diagnosis of SARS-CoV-2, which can be handled at BSL-2, has affected the transmission dynamics of COVID-19. However, new emerging and re-emerging viruses with biological flexibility classified as a Risk Group 3 agent can take advantage of epidemiological opportunities, generating the need to increase clinical diagnosis, mainly in border regions between countries. The overall objective of this project was to assure the quality of the analytical process in the diagnosis of high biological risk pathogens in Nariño by building a laboratory that includes biosafety level (BSL)-2 and (BSL)-3 containment zones. The delimitation of zones was carried out according to the Verification Tool of the National Health Institute of Colombia and following the standard requirements for the competence of testing and calibration laboratories of the International Organization for Standardization. This is achieved by harmonization of methods and equipment for effective and durable diagnostics of the large-scale spread of highly pathogenic microorganisms, employing negative-pressure containment systems and UV Systems in accordance with a finely controlled electrical system and PCR systems as new diagnostic tools. That increases laboratory capacity. Protection in BSL-3 zones will separate the handling of potentially infectious aerosols within the laboratory from the community and the environment. It will also allow the handling and inactivation of samples with suspected pathogens and the extraction of molecular material from them, allowing research with pathogens with high risks, such as SARS-CoV-2, Influenza, and syncytial virus, and malaria, among others. The diagnosis of these pathogens will be articulated across the spectrum of basic, applied, and translational research that could receive about 60 daily samples. It is expected that this project will be articulated with the health policies of neighboring countries to increase research capacity.

Keywords: medical laboratory science, SARS-CoV-2, public health surveillance, Colombia

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430 Evolutionary Advantages of Loneliness with an Agent-Based Model

Authors: David Gottlieb, Jason Yoder

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The feeling of loneliness is not uncommon in modern society, and yet, there is a fundamental lack of understanding in its origins and purpose in nature. One interpretation of loneliness is that it is a subjective experience that punishes a lack of social behavior, and thus its emergence in human evolution is seemingly tied to the survival of early human tribes. Still, a common counterintuitive response to loneliness is a state of hypervigilance, resulting in social withdrawal, which may appear maladaptive to modern society. So far, no computational model of loneliness’ effect during evolution yet exists; however, agent-based models (ABM) can be used to investigate social behavior, and applying evolution to agents’ behaviors can demonstrate selective advantages for particular behaviors. We propose an ABM where each agent contains four social behaviors, and one goal-seeking behavior, letting evolution select the best behavioral patterns for resource allocation. In our paper, we use an algorithm similar to the boid model to guide the behavior of agents, but expand the set of rules that govern their behavior. While we use cohesion, separation, and alignment for simple social movement, our expanded model adds goal-oriented behavior, which is inspired by particle swarm optimization, such that agents move relative to their personal best position. Since agents are given the ability to form connections by interacting with each other, our final behavior guides agent movement toward its social connections. Finally, we introduce a mechanism to represent a state of loneliness, which engages when an agent's perceived social involvement does not meet its expected social involvement. This enables us to investigate a minimal model of loneliness, and using evolution we attempt to elucidate its value in human survival. Agents are placed in an environment in which they must acquire resources, as their fitness is based on the total resource collected. With these rules in place, we are able to run evolution under various conditions, including resource-rich environments, and when disease is present. Our simulations indicate that there is strong selection pressure for social behavior under circumstances where there is a clear discrepancy between initial resource locations, and against social behavior when disease is present, mirroring hypervigilance. This not only provides an explanation for the emergence of loneliness, but also reflects the diversity of response to loneliness in the real world. In addition, there is evidence of a richness of social behavior when loneliness was present. By introducing just two resource locations, we observed a divergence in social motivation after agents became lonely, where one agent learned to move to the other, who was in a better resource position. The results and ongoing work from this project show that it is possible to glean insight into the evolutionary advantages of even simple mechanisms of loneliness. The model we developed has produced unexpected results and has led to more questions, such as the impact loneliness would have at a larger scale, or the effect of creating a set of rules governing interaction beyond adjacency.

Keywords: agent-based, behavior, evolution, loneliness, social

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429 Assessment of a Rapid Detection Sensor of Faecal Pollution in Freshwater

Authors: Ciprian Briciu-Burghina, Brendan Heery, Dermot Brabazon, Fiona Regan

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Good quality bathing water is a highly desirable natural resource which can provide major economic, social, and environmental benefits. Both in Ireland and Europe, such water bodies are managed under the European Directive for the management of bathing water quality (BWD). The BWD aims mainly: (i) to improve health protection for bathers by introducing stricter standards for faecal pollution assessment (E. coli, enterococci), (ii) to establish a more pro-active approach to the assessment of possible pollution risks and the management of bathing waters, and (iii) to increase public involvement and dissemination of information to the general public. Standard methods for E. coli and enterococci quantification rely on cultivation of the target organism which requires long incubation periods (from 18h to a few days). This is not ideal when immediate action is required for risk mitigation. Municipalities that oversee the bathing water quality and deploy appropriate signage have to wait for laboratory results. During this time, bathers can be exposed to pollution events and health risks. Although forecasting tools exist, they are site specific and as consequence extensive historical data is required to be effective. Another approach for early detection of faecal pollution is the use of marker enzymes. β-glucuronidase (GUS) is a widely accepted biomarker for E. coli detection in microbiological water quality control. GUS assay is particularly attractive as they are rapid, less than 4 h, easy to perform and they do not require specialised training. A method for on-site detection of GUS from environmental samples in less than 75 min was previously demonstrated. In this study, the capability of ColiSense as an early warning system for faecal pollution in freshwater is assessed. The system successfully detected GUS activity in all of the 45 freshwater samples tested. GUS activity was found to correlate linearly with E. coli (r2=0.53, N=45, p < 0.001) and enterococci (r2=0.66, N=45, p < 0.001) Although GUS is a marker for E. coli, a better correlation was obtained for enterococci. For this study water samples were collected from 5 rivers in the Dublin area over 1 month. This suggests a high diversity of pollution sources (agricultural, industrial, etc) as well as point and diffuse pollution sources were captured in the sample size. Such variety in the source of E. coli can account for different GUS activities/culturable cell and different ratios of viable but not culturable to viable culturable bacteria. A previously developed protocol for the recovery and detection of E. coli was coupled with a miniaturised fluorometer (ColiSense) and the system was assessed for the rapid detection FIB in freshwater samples. Further work will be carried out to evaluate the system’s performance on seawater samples.

Keywords: faecal pollution, β-glucuronidase (GUS), bathing water, E. coli

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428 Detection and Identification of Antibiotic Resistant Bacteria Using Infra-Red-Microscopy and Advanced Multivariate Analysis

Authors: Uraib Sharaha, Ahmad Salman, Eladio Rodriguez-Diaz, Elad Shufan, Klaris Riesenberg, Irving J. Bigio, Mahmoud Huleihel

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Antimicrobial drugs have an important role in controlling illness associated with infectious diseases in animals and humans. However, the increasing resistance of bacteria to a broad spectrum of commonly used antibiotics has become a global health-care problem. Rapid determination of antimicrobial susceptibility of a clinical isolate is often crucial for the optimal antimicrobial therapy of infected patients and in many cases can save lives. The conventional methods for susceptibility testing like disk diffusion are time-consuming and other method including E-test, genotyping are relatively expensive. Fourier transform infrared (FTIR) microscopy is rapid, safe, and low cost method that was widely and successfully used in different studies for the identification of various biological samples including bacteria. The new modern infrared (IR) spectrometers with high spectral resolution enable measuring unprecedented biochemical information from cells at the molecular level. Moreover, the development of new bioinformatics analyses combined with IR spectroscopy becomes a powerful technique, which enables the detection of structural changes associated with resistivity. The main goal of this study is to evaluate the potential of the FTIR microscopy in tandem with machine learning algorithms for rapid and reliable identification of bacterial susceptibility to antibiotics in time span of few minutes. The bacterial samples, which were identified at the species level by MALDI-TOF and examined for their susceptibility by the routine assay (micro-diffusion discs), are obtained from the bacteriology laboratories in Soroka University Medical Center (SUMC). These samples were examined by FTIR microscopy and analyzed by advanced statistical methods. Our results, based on 550 E.coli samples, were promising and showed that by using infrared spectroscopic technique together with multivariate analysis, it is possible to classify the tested bacteria into sensitive and resistant with success rate higher than 85% for eight different antibiotics. Based on these preliminary results, it is worthwhile to continue developing the FTIR microscopy technique as a rapid and reliable method for identification antibiotic susceptibility.

Keywords: antibiotics, E. coli, FTIR, multivariate analysis, susceptibility

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427 Screening for Non-hallucinogenic Neuroplastogens as Drug Candidates for the Treatment of Anxiety, Depression, and Posttraumatic Stress Disorder

Authors: Jillian M. Hagel, Joseph E. Tucker, Peter J. Facchini

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With the aim of establishing a holistic approach for the treatment of central nervous system (CNS) disorders, we are pursuing a drug development program rapidly progressing through discovery and characterization phases. The drug candidates identified in this program are referred to as neuroplastogens owing to their ability to mediate neuroplasticity, which can be beneficial to patients suffering from anxiety, depression, or posttraumatic stress disorder. These and other related neuropsychiatric conditions are associated with the onset of neuronal atrophy, which is defined as a reduction in the number and/or productivity of neurons. The stimulation of neuroplasticity results in an increase in the connectivity between neurons and promotes the restoration of healthy brain function. We have synthesized a substantial catalogue of proprietary indolethylamine derivatives based on the general structures of serotonin (5-hydroxytryptamine) and psychedelic molecules such as N,N-dimethyltryptamine (DMT) and psilocin (4-hydroxy-DMT) that function as neuroplastogens. A primary objective in our screening protocol is the identification of derivatives associated with a significant reduction in hallucination, which will allow administration of the drug at a dose that induces neuroplasticity and triggers other efficacious outcomes in the treatment of targeted CNS disorders but which does not cause a psychedelic response in the patient. Both neuroplasticity and hallucination are associated with engagement of the 5HT2A receptor, requiring drug candidates differentially coupled to these two outcomes at a molecular level. We use novel and proprietary artificial intelligence algorithms to predict the mode of binding to the 5HT2A receptor, which has been shown to correlate with the hallucinogenic response. Hallucination is tested using the mouse head-twitch response model, whereas mouse marble-burying and sucrose preference assays are used to evaluate anxiolytic and anti-depressive potential. Neuroplasticity is assays using dendritic outgrowth assays and cell-based ELISA analysis. Pharmacokinetics and additional receptor-binding analyses also contribute the selection of lead candidates. A summary of the program is presented.

Keywords: neuroplastogen, non-hallucinogenic, drug development, anxiety, depression, PTSD, indolethylamine derivatives, psychedelic-inspired, 5-HT2A receptor, computational chemistry, head-twitch response behavioural model, neurite outgrowth assay

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426 Effect of Locally Injected Mesenchymal Stem Cells on Bone Regeneration of Rat Calvaria Defects

Authors: Gileade P. Freitas, Helena B. Lopes, Alann T. P. Souza, Paula G. F. P. Oliveira, Adriana L. G. Almeida, Paulo G. Coelho, Marcio M. Beloti, Adalberto L. Rosa

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Bone tissue presents great capacity to regenerate when injured by trauma, infectious processes, or neoplasia. However, the extent of injury may exceed the inherent tissue regeneration capability demanding some kind of additional intervention. In this scenario, cell therapy has emerged as a promising alternative to treat challenging bone defects. This study aimed at evaluating the effect of local injection of bone marrow-derived mesenchymal stem cells (BM-MSCs) and adipose tissue-derived mesenchymal stem cells (AT-MSCs) on bone regeneration of rat calvaria defects. BM-MSCs and AT-MSCs were isolated and characterized by expression of surface markers; cell viability was evaluated after injection through a 21G needle. Defects of 5 mm in diameter were created in calvaria and after two weeks a single injection of BM-MSCs, AT-MSCs or vehicle-PBS without cells (Control) was carried out. Cells were tracked by bioluminescence and at 4 weeks post-injection bone formation was evaluated by micro-computed tomography (μCT) and histology, nanoindentation, and through gene expression of bone remodeling markers. The data were evaluated by one-way analysis of variance (p≤0.05). BM-MSCs and AT-MSCs presented characteristics of mesenchymal stem cells, kept viability after passing through a 21G needle and remained in the defects until day 14. In general, injection of both BM-MSCs and AT-MSCs resulted in higher bone formation compared to Control. Additionally, this bone tissue displayed elastic modulus and hardness similar to the pristine calvaria bone. The expression of all evaluated genes involved in bone formation was upregulated in bone tissue formed by BM-MSCs compared to AT-MSCs while genes involved in bone resorption were upregulated in AT-MSCs-formed bone. We show that cell therapy based on the local injection of BM-MSCs or AT-MSCs is effective in delivering viable cells that displayed local engraftment and induced a significant improvement in bone healing. Despite differences in the molecular cues observed between BM-MSCs and AT-MSCs, both cells were capable of forming bone tissue at comparable amounts and properties. These findings may drive cell therapy approaches toward the complete bone regeneration of challenging sites.

Keywords: cell therapy, mesenchymal stem cells, bone repair, cell culture

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425 The Role of Islamic Finance and Socioeconomic Factors in Financial Inclusion: A Cross Country Comparison

Authors: Allya Koesoema, Arni Ariani

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While religion is only a very minor factor contributing to financial exclusion in most countries, the World Bank 2014 Global Financial Development Report highlighted it as a significant barrier for having a financial account in some Muslim majority countries. This is in part due to the perceived incompatibility between traditional financial institutions practices and Islamic finance principles. In these cases, the development of financial institutions and products that are compatible with the principles of Islamic finance may act as an important lever to increasing formal account ownership. However, there is significant diversity in the relationship between a country’s proportion of Muslim population and its level of financial inclusion. This paper combines data taken from the Global Findex Database, World Development Indicators, and the Pew Research Center to quantitatively explore the relationship between individual and country level religious and socioeconomic factor to financial inclusion. Results from regression analyses show a complex relationship between financial inclusion and religion-related factors in the population both on the individual and country level. Consistent with prior literature, on average the percentage of Islamic population positively correlates with the proportion of unbanked populations who cites religious reasons as a barrier to getting an account. However, its impact varies across several variables. First, a deeper look into countries’ religious composition reveals that the average negative impact of a large Muslim population is not as strong in more religiously diverse countries and less religious countries. Second, on the individual level, among the unbanked, the poorest quintile, least educated, older and the female populations are comparatively more likely to not have an account because of religious reason. Results also show indications that in this case, informal mechanisms partially substitute formal financial inclusion, as indicated by the propensity to borrow from family and friends. The individual level findings are important because the demographic groups that are more likely to cite religious reasons as barriers to formal financial inclusion are also generally perceived to be more vulnerable socially and economically and may need targeted attention. Finally, the number of Islamic financial institutions in a particular country is negatively correlated to the propensity of religious reasons as a barrier to financial inclusion. Importantly, the number of financial institutions in a country also mitigates the negative impact of the proportion of Muslim population, low education and individual age to formal financial inclusion. These results point to the potential importance of Islamic Finance Institutions in increasing global financial inclusion, and highlight the potential importance of looking beyond the proportion of Muslim population to other underlying institutional and socioeconomic factor in maximizing its impact.

Keywords: cross country comparison, financial inclusion, Islamic banking and finance, quantitative methods, socioeconomic factors

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424 Architectural Wind Data Maps Using an Array of Wireless Connected Anemometers

Authors: D. Serero, L. Couton, J. D. Parisse, R. Leroy

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In urban planning, an increasing number of cities require wind analysis to verify comfort of public spaces and around buildings. These studies are made using computer fluid dynamic simulation (CFD). However, this technique is often based on wind information taken from meteorological stations located at several kilometers of the spot of analysis. The approximated input data on project surroundings produces unprecise results for this type of analysis. They can only be used to get general behavior of wind in a zone but not to evaluate precise wind speed. This paper presents another approach to this problem, based on collecting wind data and generating an urban wind cartography using connected ultrasound anemometers. They are wireless devices that send immediate data on wind to a remote server. Assembled in array, these devices generate geo-localized data on wind such as speed, temperature, pressure and allow us to compare wind behavior on a specific site or building. These Netatmo-type anemometers communicate by wifi with central equipment, which shares data acquired by a wide variety of devices such as wind speed, indoor and outdoor temperature, rainfall, and sunshine. Beside its precision, this method extracts geo-localized data on any type of site that can be feedback looped in the architectural design of a building or a public place. Furthermore, this method allows a precise calibration of a virtual wind tunnel using numerical aeraulic simulations (like STAR CCM + software) and then to develop the complete volumetric model of wind behavior over a roof area or an entire city block. The paper showcases connected ultrasonic anemometers, which were implanted for an 18 months survey on four study sites in the Grand Paris region. This case study focuses on Paris as an urban environment with multiple historical layers whose diversity of typology and buildings allows considering different ways of capturing wind energy. The objective of this approach is to categorize the different types of wind in urban areas. This, particularly the identification of the minimum and maximum wind spectrum, helps define the choice and performance of wind energy capturing devices that could be implanted there. The localization on the roof of a building, the type of wind, the altimetry of the device in relation to the levels of the roofs, the potential nuisances generated. The method allows identifying the characteristics of wind turbines in order to maximize their performance in an urban site with turbulent wind.

Keywords: computer fluid dynamic simulation in urban environment, wind energy harvesting devices, net-zero energy building, urban wind behavior simulation, advanced building skin design methodology

Procedia PDF Downloads 84
423 Effective Layer-by-layer Chemical Grafting of a Reactive Oxazoline Polymer and MWCNTs onto Carbon Fibers for Enhancing Mechanical Properties of Composites using Polystyrene as a Model Thermoplastic Matrix

Authors: Ryoma Tokonami, Teruya Goto, Tatsuhiro Takahashi,

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For enhancing the mechanical property ofcarbon fiber reinforced plastic (CFRP), the surface modification of carbon fiber (CF) by multi-walled carbon nanotube (MWCNT) has received considerable attention using direct MWCNT growth on CF with a catalysis, MWCNT electrophoresis, and layer-by-layer of MWCNT with reactive polymers, etc. Among above approaches, the layer-by-layer method is the simplest process, however, the amount of MWCNTs on CF is very little, resulting in the small amount of improvement of the mechanical property of the composite. The remaining amount of MWCNT on CF after melt mixing of CF (short fiber) with thermoplastic matrix polymer was not examined clearly in the former studies. The present research aims to propose an effective layer-by-layer chemical grafting of a highly reactive oxazoline polymer, which has not been used before, and MWCNTs onto CF using the highly reactivity of oxazoline and COOH on the surface of CF and MWCNTs.With layer-by-layer method, the first uniform chemically bonded mono molecular layer on carbon fiber was formed by chemical surface reaction of carbon fiber, a reactive oxazoline polymer solution between COOH of carbon fiber and oxazoline. The second chemically bonded uniform layer of MWCNTs on the first layer was prepared through the first layer coated carbon fiber in MWCNT dispersion solution by chemical reaction between oxazoline and COOH of MWCNTs. The quantitative analysis of MWCNTs on carbon fiber was performed, showing 0.44 wt.% of MWCNTs based on carbon fiber, which is much larger amount compared with the former studies in layer-by-layer method. In addition, MWCNTs were also observed uniform coating on carbon fiber by scanning electron micrograph (SEM). Carbon fiber composites were prepared by melting mixing using polystyrene (PS) as a thermoplastic matrix because of easy removal of PS by solvent for additional analysis, resulting the 20% of enhancement of tensile strength and modulus by tensile strength test. It was confirmed bySEM the layer-by-layer structure on carbon fibers were remained after the melt mixing by removing PS with a solvent. As a conclusion, the effectiveness for the enhancement of the mechanical properties of CF(short fiber)/PS composite using the highly reactive oxazoline polymer for the first layer and MWCNT for the second layer, which act as the physical anchor, was demonstrated.

Keywords: interface, layer-by-layer, multi walled carbon nanotubes (MWCNTs), oxazoline

Procedia PDF Downloads 182
422 How Does Paradoxical Leadership Enhance Organizational Success?

Authors: Wageeh A. Nafei

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This paper explores the role of Paradoxical Leadership (PL) in enhancing Organizational Success (OS) at private hospitals in Egypt. Based on the collected data from employees in private hospitals (doctors, nursing staff, and administrative staff). The researcher has adopted a sampling method to collect data for the study. The appropriate statistical methods, such as Alpha Correlation Coefficient (ACC), Confirmatory Factor Analysis (CFA), and Multiple Regression Analysis (MRA), are used to analyze the data and test the hypotheses. The research has reached a number of results, the most important of which are (1) there is a statistical relationship between the independent variable represented by PL and the dependent variable represented by Organizational Success (OS). The paradoxical leader encourages employees to express their opinions and builds a work environment characterized by flexibility and independence. Also, the paradoxical leader works to support specialized work teams, which leads to the creation of new ideas, on the one hand, and contributes to the achievement of outstanding performance on the other hand. (2) the mentality of the paradoxical leader is flexible and capable of absorbing all suggestions from all employees. Also, the paradoxical leader is interested in enhancing cooperation among them and provides an opportunity to transfer experience and increase knowledge-sharing. Also, the sharing of knowledge creates the necessary diversity that helps the organization to obtain rich external information and enables the organization to deal with a rapidly changing environment. (3) The PL approach helps in facing the paradoxical demands of employees. A paradoxical leader plays an important role in reducing the feeling of instability in the work environment and lack of job security, reducing negative feelings for employees, restoring balance in the work environment, improving the well-being of employees, and increasing the degree of job satisfaction of employees in the organization. The study referred to a number of recommendations, the most important of which are (1) the leaders of the organizations must listen to the views of employees and their needs and move away from the official method of control. The leader should give sufficient freedom to employees to participate in decision-making and maintain enough space among them. The treatment between the leaders and employees must be based on friendliness, (2) the need for organizational leaders to pay attention to sharing knowledge among employees through training courses. The leader should make sure that every information provided by the employee is valuable and useful, which can be used to solve a problem that may face his/her colleagues at work, (3) the need for organizational leaders to pay attention to sharing knowledge among employees through brainstorming sessions. The leader should ensure that employees obtain knowledge from their colleagues and share ideas and information among them. This is in addition to motivating employees to complete their work in a new creative way, which leads to employees’ not feeling bored of repeating the same routine procedures in the organization.

Keywords: paradoxical leadership, organizational success, human resourece, management

Procedia PDF Downloads 44
421 An Impregnated Active Layer Mode of Solution Combustion Synthesis as a Tool for the Solution Combustion Mechanism Investigation

Authors: Zhanna Yermekova, Sergey Roslyakov

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Solution combustion synthesis (SCS) is the unique method which multiple times has proved itself as an effective and efficient approach for the versatile synthesis of a variety of materials. It has significant advantages such as relatively simple handling process, high rates of product synthesis, mixing of the precursors on a molecular level, and fabrication of the nanoproducts as a result. Nowadays, an overwhelming majority of solution combustion investigations performed through the volume combustion synthesis (VCS) where the entire liquid precursor is heated until the combustion self-initiates throughout the volume. Less amount of the experiments devoted to the steady-state self-propagating mode of SCS. Under the beforementioned regime, the precursor solution is dried until the gel-like media, and later on, the gel substance is locally ignited. In such a case, a combustion wave propagates in a self-sustaining mode as in conventional solid combustion synthesis. Even less attention is given to the impregnated active layer (IAL) mode of solution combustion. An IAL approach to the synthesis is implying that the solution combustion of the precursors should be initiated on the surface of the third chemical or inside the third substance. This work is aiming to emphasize an underestimated role of the impregnated active layer mode of the solution combustion synthesis for the fundamental studies of the combustion mechanisms. It also serves the purpose of popularizing the technical terms and clarifying the difference between them. In order to do so, the solution combustion synthesis of γ-FeNi (PDF#47-1417) alloy has been accomplished within short (seconds) one-step reaction of metal precursors with hexamethylenetetramine (HTMA) fuel. An idea of the special role of the Ni in a process of alloy formation was suggested and confirmed with the particularly organized set of experiments. The first set of experiments were conducted in a conventional steady-state self-propagating mode of SCS. An alloy was synthesized as a single monophasic product. In two other experiments, the synthesis was divided into two independent processes which are possible under the IAL mode of solution combustion. The sequence of the process was changed according to the equations which are describing an Experiment A and B below: Experiment A: Step 1. Fe(NO₃)₃*9H₂O + HMTA = FeO + gas products; Step 2. FeO + Ni(NO₃)₂*6H₂O + HMTA = Ni + FeO + gas products; Experiment B: Step 1. Ni(NO₃)₂*6H₂O + HMTA = Ni + gas products; Step 2. Ni + Fe(NO₃)₃*9H₂O + HMTA = Fe₃Ni₂+ traces (Ni + FeO). Based on the IAL experiment results, one can see that combustion of the Fe(NO₃)₃9H₂O on the surface of the Ni is leading to the alloy formation while presence of the already formed FeO does not affect the Ni(NO₃)₂*6H₂O + HMTA reaction in any way and Ni is the main product of the synthesis.

Keywords: alloy, hexamethylenetetramine, impregnated active layer mode, mechanism, solution combustion synthesis

Procedia PDF Downloads 115
420 Road Systems as Environmental Barriers: An Overview of Roadways in Their Function as Fences for Wildlife Movement

Authors: Rachael Bentley, Callahan Gergen, Brodie Thiede

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Roadways have a significant impact on the environment in so far as they function as barriers to wildlife movement, both through road mortality and through resultant road avoidance. Roads have an im-mense presence worldwide, and it is predicted to increase substantially in the next thirty years. As roadways become even more common, it is important to consider their environmental impact, and to mitigate the negative effects which they have on wildlife and wildlife mobility. In a thorough analysis of several related studies, a common conclusion was that roads cause habitat fragmentation, which can lead split populations to evolve differently, for better or for worse. Though some populations adapted positively to roadways, becoming more resistant to road mortality, and more tolerant to noise and chemical contamination, many others experienced maladaptation, either due to chemical contamination in and around their environment, or because of genetic mutations from inbreeding when their population was fragmented too substantially to support a large enough group for healthy genetic exchange. Large mammals were especially susceptible to maladaptation from inbreed-ing, as they require larger areas to roam and therefore require even more space to sustain a healthy population. Regardless of whether a species evolved positively or negatively as a result of their proximity to a road, animals tended to avoid roads, making the genetic diversity from habitat fragmentation an exceedingly prevalent issue in the larger discussion of road ecology. Additionally, the consideration of solu-tions, such as overpasses and underpasses, is crucial to ensuring the long term survival of many wildlife populations. In studies addressing the effectiveness of overpasses and underpasses, it seemed as though animals adjusted well to these sorts of solutions, but strategic place-ment, as well as proper sizing, proper height, shelter from road noise, and other considerations were important in construction. When an underpass or overpass was well-built and well-shielded from human activity, animals’ usage of the structure increased significantly throughout its first five years, thus reconnecting previously divided populations. Still, these structures are costly and they are often unable to fully address certain issues such as light, noise, and contaminants from vehicles. Therefore, the need for further discussion of new, crea-tive solutions remains paramount. Roads are one of the most consistent and prominent features of today’s landscape, but their environmental impacts are largely overlooked. While roads are useful for connecting people, they divide landscapes and animal habitats. Therefore, further research and investment in possible solutions is necessary to mitigate the negative effects which roads have on wildlife mobility and to pre-vent issues from resultant habitat fragmentation.

Keywords: fences, habitat fragmentation, roadways, wildlife mobility

Procedia PDF Downloads 151
419 Expression of miRNA 335 in Gall Bladder Cancer: A Correlative Study

Authors: Naseem Fatima, A. N. Srivastava, Tasleem Raza, Vijay Kumar

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Introduction: Carcinoma gallbladder is third most common gastrointestinal lethal disease with the highest incidence and mortality rate among women in Northern India. Scientists have found several risk factors that make a person more likely to develop gallbladder cancer; among these risk factors, deregulation of miRNAs has been demonstrated to be one of the most crucial factors. The changes in the expression of specific miRNA genes result in the control of inflammation, cell cycle regulation, stress response, proliferation, differentiation, apoptosis and invasion thus mediate the process in tumorgenesis. The aim of this study was to investigate the role of MiRNA-335 and may as a molecular marker in early detection of gallbladder cancer in suspected cases. Material and Methods: A total of 20 consecutive patients with gallbladder cancer aged between 30-75 years were registered for the study. Total RNA was extracted from tissue by using the mirVANA MiRNA isolation Kit according to the manufacturer’s protocol. The MiRNA- 335 and U6 snRNA-specific cDNA were reverse-transcribed from total RNA using Taqman microRNA reverse-transcription kit according to the manufacturer’s protocol. TaqMan MiRNA probes hsa-miR-335 and Taqman Master Mix without AmpEase UNG, Individual real-time PCR assays were performed in a 20 μL reaction volume on a Real-Time PCR system (Applied Biosystems StepOnePlus™) to detect MiRNA-335 expression in tissue. Relative quantification of target MiRNA expression was evaluated using the comparative cycle threshold (CT) method. The correlation was done in between cycle threshold (CT Value) of target MiRNA in gallbladder cancer with respect to non-cancerous Cholelithiasis gallbladder. Each sample was examined in triplicate. The Newman-Keuls Multiple Comparison Test was used to determine the expression of miR-335. Results: MiRNA335 was found to be significantly downregulated in the gallbladder cancer tissue (P<0.001), when compared with non-cancerous Cholelithiasis gallbladder cases. Out of 20 cases, 75% showed reduced expression of MiRNA335, were at last stage of disease with low overall survival rate and remaining 25% were showed up-regulated expression of MiRNA335 with high survival rate. Conclusion: The present study showed that reduced expression of MiRNA335 is associated with the advancement of the disease, and its deregulation may provide important clues to understanding it as a prognostic marker and opportunities for future research.

Keywords: carcinoma gallbladder, downregulation, MiRNA-335, RT-PCR assay

Procedia PDF Downloads 343
418 Hydrogen Sulfide Releasing Ibuprofen Derivative Can Protect Heart After Ischemia-Reperfusion

Authors: Virag Vass, Ilona Bereczki, Erzsebet Szabo, Nora Debreczeni, Aniko Borbas, Pal Herczegh, Arpad Tosaki

Abstract:

Hydrogen sulfide (H₂S) is a toxic gas, but it is produced by certain tissues in a small quantity. According to earlier studies, ibuprofen and H₂S has a protective effect against damaging heart tissue caused by ischemia-reperfusion. Recently, we have been investigating the effect of a new water-soluble H₂S releasing ibuprofen molecule administered after artificially generated ischemia-reperfusion on isolated rat hearts. The H₂S releasing property of the new ibuprofen derivative was investigated in vitro in medium derived from heart endothelial cell isolation at two concentrations. The ex vivo examinations were carried out on rat hearts. Rats were anesthetized with an intraperitoneal injection of ketamine, xylazine, and heparin. After thoracotomy, hearts were excised and placed into ice-cold perfusion buffer. Perfusion of hearts was conducted in Langendorff mode via the cannulated aorta. In our experiments, we studied the dose-effect of the H₂S releasing molecule in Langendorff-perfused hearts with the application of gradually increasing concentration of the compound (0- 20 µM). The H₂S releasing ibuprofen derivative was applied before the ischemia for 10 minutes. H₂S concentration was measured with an H₂S detecting electrochemical sensor from the coronary effluent solution. The 10 µM concentration was chosen for further experiments when the treatment with this solution was occurred after the ischemia. The release of H₂S is occurred by the hydrolyzing enzymes that are present in the heart endothelial cells. The protective effect of the new H₂S releasing ibuprofen molecule can be confirmed by the infarct sizes of hearts using the Triphenyl-tetrazolium chloride (TTC) staining method. Furthermore, we aimed to define the effect of the H₂S releasing ibuprofen derivative on autophagic and apoptotic processes in damaged hearts after investigating the molecular markers of these events by western blotting and immunohistochemistry techniques. Our further studies will include the examination of LC3I/II, p62, Beclin1, caspase-3, and other apoptotic molecules. We hope that confirming the protective effect of new H₂S releasing ibuprofen molecule will open a new possibility for the development of more effective cardioprotective agents with exerting fewer side effects. Acknowledgment: This study was supported by the grants of NKFIH- K-124719 and the European Union and the State of Hungary co- financed by the European Social Fund in the framework of GINOP- 2.3.2-15-2016-00043.

Keywords: autophagy, hydrogen sulfide, ibuprofen, ischemia, reperfusion

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417 Selection of Suitable Reference Genes for Assessing Endurance Related Traits in a Native Pony Breed of Zanskar at High Altitude

Authors: Prince Vivek, Vijay K. Bharti, Manishi Mukesh, Ankita Sharma, Om Prakash Chaurasia, Bhuvnesh Kumar

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High performance of endurance in equid requires adaptive changes involving physio-biochemical, and molecular responses in an attempt to regain homeostasis. We hypothesized that the identification of the suitable reference genes might be considered for assessing of endurance related traits in pony at high altitude and may ensure for individuals struggling to potent endurance trait in ponies at high altitude. A total of 12 mares of ponies, Zanskar breed, were divided into three groups, group-A (without load), group-B, (60 Kg) and group-C (80 Kg) on backpack loads were subjected to a load carry protocol, on a steep climb of 4 km uphill, and of gravel, uneven rocky surface track at an altitude of 3292 m to 3500 m (endpoint). Blood was collected before and immediately after the load carry on sodium heparin anticoagulant, and the peripheral blood mononuclear cell was separated for total RNA isolation and thereafter cDNA synthesis. Real time-PCR reactions were carried out to evaluate the mRNAs expression profile of a panel of putative internal control genes (ICGs), related to different functional classes, namely glyceraldehyde 3-phosphate dehydrogenase (GAPDH), β₂ microglobulin (β₂M), β-actin (ACTB), ribosomal protein 18 (RS18), hypoxanthine-guanine phosophoribosyltransferase (HPRT), ubiquitin B (UBB), ribosomal protein L32 (RPL32), transferrin receptor protein (TFRC), succinate dehydrogenase complex subunit A (SDHA) for normalizing the real-time quantitative polymerase chain reaction (qPCR) data of native pony’s. Three different algorithms, geNorm, NormFinder, and BestKeeper software, were used to evaluate the stability of reference genes. The result showed that GAPDH was best stable gene and stability value for the best combination of two genes was observed TFRC and β₂M. In conclusion, the geometric mean of GAPDH, TFRC and β₂M might be used for accurate normalization of transcriptional data for assessing endurance related traits in Zanskar ponies during load carrying.

Keywords: endurance exercise, ubiquitin B (UBB), β₂ microglobulin (β₂M), high altitude, Zanskar ponies, reference gene

Procedia PDF Downloads 119
416 Evaluation of Yield and Yield Components of Malaysian Palm Oil Board-Senegal Oil Palm Germplasm Using Multivariate Tools

Authors: Khin Aye Myint, Mohd Rafii Yusop, Mohd Yusoff Abd Samad, Shairul Izan Ramlee, Mohd Din Amiruddin, Zulkifli Yaakub

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The narrow base of genetic is the main obstacle of breeding and genetic improvement in oil palm industry. In order to broaden the genetic bases, the Malaysian Palm Oil Board has been extensively collected wild germplasm from its original area of 11 African countries which are Nigeria, Senegal, Gambia, Guinea, Sierra Leone, Ghana, Cameroon, Zaire, Angola, Madagascar, and Tanzania. The germplasm collections were established and maintained as a field gene bank in Malaysian Palm Oil Board (MPOB) Research Station in Kluang, Johor, Malaysia to conserve a wide range of oil palm genetic resources for genetic improvement of Malaysian oil palm industry. Therefore, assessing the performance and genetic diversity of the wild materials is very important for understanding the genetic structure of natural oil palm population and to explore genetic resources. Principal component analysis (PCA) and Cluster analysis are very efficient multivariate tools in the evaluation of genetic variation of germplasm and have been applied in many crops. In this study, eight populations of MPOB-Senegal oil palm germplasm were studied to explore the genetic variation pattern using PCA and cluster analysis. A total of 20 yield and yield component traits were used to analyze PCA and Ward’s clustering using SAS 9.4 version software. The first four principal components which have eigenvalue >1 accounted for 93% of total variation with the value of 44%, 19%, 18% and 12% respectively for each principal component. PC1 showed highest positive correlation with fresh fruit bunch (0.315), bunch number (0.321), oil yield (0.317), kernel yield (0.326), total economic product (0.324), and total oil (0.324) while PC 2 has the largest positive association with oil to wet mesocarp (0.397) and oil to fruit (0.458). The oil palm population were grouped into four distinct clusters based on 20 evaluated traits, this imply that high genetic variation existed in among the germplasm. Cluster 1 contains two populations which are SEN 12 and SEN 10, while cluster 2 has only one population of SEN 3. Cluster 3 consists of three populations which are SEN 4, SEN 6, and SEN 7 while SEN 2 and SEN 5 were grouped in cluster 4. Cluster 4 showed the highest mean value of fresh fruit bunch, bunch number, oil yield, kernel yield, total economic product, and total oil and Cluster 1 was characterized by high oil to wet mesocarp, and oil to fruit. The desired traits that have the largest positive correlation on extracted PCs could be utilized for the improvement of oil palm breeding program. The populations from different clusters with the highest cluster means could be used for hybridization. The information from this study can be utilized for effective conservation and selection of the MPOB-Senegal oil palm germplasm for the future breeding program.

Keywords: cluster analysis, genetic variability, germplasm, oil palm, principal component analysis

Procedia PDF Downloads 149
415 N-Glycosylation in the Green Microalgae Chlamydomonas reinhardtii

Authors: Pierre-Louis Lucas, Corinne Loutelier-Bourhis, Narimane Mati-Baouche, Philippe Chan Tchi-Song, Patrice Lerouge, Elodie Mathieu-Rivet, Muriel Bardor

Abstract:

N-glycosylation is a post-translational modification taking place in the Endoplasmic Reticulum and the Golgi apparatus where defined glycan features are added on protein in a very specific sequence Asn-X-Thr/Ser/Cys were X can be any amino acid except proline. Because it is well-established that those N-glycans play a critical role in protein biological activity, protein half-life and that a different N-glycan structure may induce an immune response, they are very important in Biopharmaceuticals which are mainly glycoproteins bearing N-glycans. From now, most of the biopharmaceuticals are produced by mammalian cells like Chinese Hamster Ovary cells (CHO) for their N-glycosylation similar to the human, but due to the high production costs, several other species are investigated as the possible alternative system. In this purpose, the green microalgae Chlamydomonas reinhardtii was investigated as the potential production system for Biopharmaceuticals. This choice was influenced by the facts that C. reinhardtii is a well-study microalgae which is growing fast with a lot of molecular biology tools available. This organism is also producing N-glycan on its endogenous proteins. However, the analysis of the N-glycan structure of this microalgae has revealed some differences as compared to the human. Rather than in Human where the glycans are processed by key enzymes called N-acetylglucosaminyltransferase I and II (GnTI and GnTII) adding GlcNAc residue to form a GlcNAc₂Man₃GlcNAc₂ core N-glycan, C. reinhardtii lacks those two enzymes and possess a GnTI independent glycosylation pathway. Moreover, some enzymes like xylosyltransferases and methyltransferases not present in human are supposed to act on the glycans of C. reinhardtii. Furthermore, the recent structural study by mass spectrometry shows that the N-glycosylation precursor supposed to be conserved in almost all eukaryotic cells results in a linear Man₅GlcNAc₂ rather than a branched one in C. reinhardtii. In this work, we will discuss the new released MS information upon C. reinhardtii N-glycan structure and their impact on our attempt to modify the glycan in a Human manner. Two strategies will be discussed. The first one consisted in the study of Xylosyltransferase insertional mutants from the CLIP library in order to remove xyloses from the N-glycans. The second will go further in the humanization by transforming the microalgae with the exogenous gene from Toxoplasma gondii having an activity similar to GnTI and GnTII with the aim to synthesize GlcNAc₂Man₃GlcNAc₂ in C. reinhardtii.

Keywords: Chlamydomonas reinhardtii, N-glycosylation, glycosyltransferase, mass spectrometry, humanization

Procedia PDF Downloads 155
414 Evaluation of Antioxidant Activity and Total Phenolic Content of Lens Esculenta Moench, Seeds

Authors: Vivek Kumar Gupta, Kripi Vohra, Monika Gupta

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Pulses have been a vital ingredient of the balanced human diet in India. Lentil (Lens culinaris Medikus or Lens esculenta Moench.) is a common legume known since biblical times. Lentil seeds, with or without hulls, are cooked as dhal and this has been the main dish for millennia in the South Asian region. Oxidative stress can damage lipids, proteins, enzymes, carbohydrates and DNA in cells and tissues, resulting in membrane damage, fragmentation or random cross linking of molecules like DNA, enzymes and structural proteins and even lead to cell death induced by DNA fragmentation and lipid peroxidation. These consequences of oxidative stress construct the molecular basis in the development of cancer, neurodegenerative disorders, cardiovascular diseases, diabetes and autoimmune. The aim of the present work is to assess the antioxidant potential of the peteroleum ether, acetone, methanol and water extract of the Lens esculenta seeds. In vitro antioxidant assessment of the extracts was carried out using 1,1-diphenyl-2-picryl hydrazyl (DPPH) radical scavenging activity, hydroxyl radical scavenging activity, reducing power assay. The quantitative estimation of total phenolic content, total flavonoid content in extracts and in plant material, total saponin content, total alkaloid content, crude fibre content, total volatile content, fat content and mucilage content in drug material was also carried out. Though all the extracts exhibited dose dependent reducing power activity the acetone extract was found to possess significant hydrogen donating ability in DPPH (45.83%-93.13%) and hydroxyl radical scavenging system (28.7%-46.41%) than the peteroleum ether, methanol and water extracts. Total phenolic content in the acetone and methanol extract was found to be 608 and 188 mg gallic acid equivalent of phenol/g of sample respectively. Total flavonoid content of acetone and methanol extract was found to be 128 and 30.6 mg quercetin equivalent/g of sample respectively. It is evident that acetone extract of Lentil seeds possess high levels of polyphenolics and flavonoids that could be utilized as antioxidants and neutraceuticals.

Keywords: antioxidant, flavanoids, Lens esculenta, polyphenols

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413 Bedouin of Silicon Wadi: A Case Study Analysis of the Multi-Level Perspectives and Factors Affecting Bedouin Entrepreneurialism as Obstacles to Entry into the Israeli High-Tech Industry

Authors: Frazer G. Thompson

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Israel is a nation of cultural and historical diversity, yet the success factors for a modern Bedouin-Arab high-tech entrepreneur seem to be different from those of other Jewish-Israeli citizens. The purpose of this descriptive narrative case study is to explore how an Arab-Israeli all Negev-Bedouin technology company has succeeded in the Israeli high-tech industry by utilizing technology and engineering career opportunities available to Bedouin youth for ‘Sadel Tech,’ at Be’er-Sheva, the Negev, Israel. Methods: The strategy of inquiry seeks to explore real-life contextual understandings, multi-level perspectives, and the cultural influences of personal, community, educational, and entrepreneurial factors. The research methodology includes in-depth one-on-one interviews, focus group sessions, and overt observation to explore the meaning and understanding of the constructs toward determining the effect all or a few of the elements may have on the overall success factors of the company. Results: Study results indicate that the state-run educational system in Israel fails to adequately integrate important aspects of Bedouin culture into the learning environment. However, Bedouin entrepreneurs are finding ways to compensate for these inadequacies by utilizing non-traditional methods of teaching, learning, and doing business. Government incentives for Bedouin start-ups are also recognized as contributors. Employees of Sadel live and work in the Negev, the Gaza Strip, and the West Bank, further informing the study that the traditions of tribal etiquette continue to contribute to modern Bedouin-Arab business culture. Conclusion: Bedouin's business success in Israel is a multi-dimensional concept. While cultural acumen plays a prominent and unique role for both Arab-Israelis and Jewish-Israelis in economic and entrepreneurial pursuits, the marginalization of the Bedouin continues to contribute to the lack of educational and professional opportunities for Bedouin in Israel. Although recognized as important at the government level, programs necessary to implement the infrastructure required to support Bedouin entrepreneurship in Israel remain infantile. The Israeli Government is providing opportunities through grants and other incentives for Bedouin entrepreneurial start-ups, indicating that Israel has recognized the impact of this growing demographic. However, although many Bedouin graduates from University each year with advanced degrees, opportunities for Bedouin within the Israeli high-tech sector remain scarce.

Keywords: Bedouin education, Bedouin entrepreneur, economic anthropology, ethnic business opportunities, Israeli tech, Silicon Wadi

Procedia PDF Downloads 102
412 Roads and Agriculture: Impacts of Connectivity in Peru

Authors: Julio Aguirre, Yohnny Campana, Elmer Guerrero, Daniel De La Torre Ugarte

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A well-developed transportation network is a necessary condition for a country to derive full benefits from good trade and macroeconomic policies. Road infrastructure plays a key role in the economic development of rural areas of developing countries; where agriculture is the main economic activity. The ability to move agricultural production from the place of production to the market, and then to the place of consumption, greatly influence the economic value of farming activities, and of the resources involved in the production process, i.e., labor and land. Consequently, investment in transportation networks contributes to enhance or overcome the natural advantages or disadvantages that topography and location have imposed over the agricultural sector. This is of particular importance when dealing with countries, like Peru, with a great topographic diversity. The objective of this research is to estimate the impacts of road infrastructure on the performance of the agricultural sector. Specific variables of interest are changes in travel time, shifts of production for self-consumption to production for the market, changes in farmers income, and impacts on the diversification of the agricultural sector. In the study, a cross-section model with instrumental variables is the central methodological instrument. The data is obtained from agricultural and transport geo-referenced databases, and the instrumental variable specification utilized is based on the Kruskal algorithm. The results show that the expansion of road connectivity reduced farmers' travel time by an average of 3.1 hours and the proportion of output sold in the market increases by up to 40 percentage points. The increase in connectivity has an unexpected increase in the districts index of diversification of agricultural production. The results are robust to the inclusion of year and region fixed-effects, and to control for geography (i.e., slope and altitude), population variables, and mining activity. Other results are also very eloquent. For example, a clear positive impact can be seen in access to local markets, but this does not necessarily correlate with an increase in the production of the sector. This can be explained by the fact that agricultural development not only requires provision of roads but additional complementary infrastructure and investments intended to provide the necessary conditions so that producers can offer quality products (improved management practices, timely maintenance of irrigation infrastructure, transparent management of water rights, among other factors). Therefore, complementary public goods are needed to enhance the effects of roads on the welfare of the population, beyond enabling them to increase their access to markets.

Keywords: agriculture devolepment, market access, road connectivity, regional development

Procedia PDF Downloads 182
411 CRISPR/Cas9 Based Gene Stacking in Plants for Virus Resistance Using Site-Specific Recombinases

Authors: Sabin Aslam, Sultan Habibullah Khan, James G. Thomson, Abhaya M. Dandekar

Abstract:

Losses due to viral diseases are posing a serious threat to crop production. A quick breakdown of resistance to viruses like Cotton Leaf Curl Virus (CLCuV) demands the application of a proficient technology to engineer durable resistance. Gene stacking has recently emerged as a potential approach for integrating multiple genes in crop plants. In the present study, recombinase technology has been used for site-specific gene stacking. A target vector (pG-Rec) was designed for engineering a predetermined specific site in the plant genome whereby genes can be stacked repeatedly. Using Agrobacterium-mediated transformation, the pG-Rec was transformed into Coker-312 along with Nicotiana tabacum L. cv. Xanthi and Nicotiana benthamiana. The transgene analysis of target lines was conducted through junction PCR. The transgene positive target lines were used for further transformations to site-specifically stack two genes of interest using Bxb1 and PhiC31 recombinases. In the first instance, Cas9 driven by multiplex gRNAs (for Rep gene of CLCuV) was site-specifically integrated into the target lines and determined by the junction PCR and real-time PCR. The resulting plants were subsequently used to stack the second gene of interest (AVP3 gene from Arabidopsis for enhancing cotton plant growth). The addition of the genes is simultaneously achieved with the removal of marker genes for recycling with the next round of gene stacking. Consequently, transgenic marker-free plants were produced with two genes stacked at the specific site. These transgenic plants can be potential germplasm to introduce resistance against various strains of cotton leaf curl virus (CLCuV) and abiotic stresses. The results of the research demonstrate gene stacking in crop plants, a technology that can be used to introduce multiple genes sequentially at predefined genomic sites. The current climate change scenario highlights the use of such technologies so that gigantic environmental issues can be tackled by several traits in a single step. After evaluating virus resistance in the resulting plants, the lines can be a primer to initiate stacking of further genes in Cotton for other traits as well as molecular breeding with elite cotton lines.

Keywords: cotton, CRISPR/Cas9, gene stacking, genome editing, recombinases

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410 Comparison of Different Methods of Microorganism's Identification from a Copper Mining in Pará, Brazil

Authors: Louise H. Gracioso, Marcela P.G. Baltazar, Ingrid R. Avanzi, Bruno Karolski, Luciana J. Gimenes, Claudio O. Nascimento, Elen A. Perpetuo

Abstract:

Introduction: Higher copper concentrations promote a selection pressure on organisms such as plants, fungi and bacteria, which allows surviving only the resistant organisms to the contaminated site. This selective pressure keeps only the organisms most resistant to a specific condition and subsequently increases their bioremediation potential. Despite the bacteria importance for biosphere maintenance, it is estimated that only a small fraction living microbial species has been described and characterized. Due to the molecular biology development, tools based on analysis 16S ribosomal RNA or another specific gene are making a new scenario for the characterization studies and identification of microorganisms in the environment. News identification of microorganisms methods have also emerged like Biotyper (MALDI / TOF), this method mass spectrometry is subject to the recognition of spectroscopic patterns of conserved and features proteins for different microbial species. In view of this, this study aimed to isolate bacteria resistant to copper present in a Copper Processing Area (Sossego Mine, Canaan, PA) and identifies them in two different methods: Recent (spectrometry mass) and conventional. This work aimed to use them for a future bioremediation of this Mining. Material and Methods: Samples were collected at fifteen different sites of five periods of times. Microorganisms were isolated from mining wastes by culture enrichment technique; this procedure was repeated 4 times. The isolates were inoculated into MJS medium containing different concentrations of chloride copper (1mM, 2.5mM, 5mM, 7.5mM and 10 mM) and incubated in plates for 72 h at 28 ºC. These isolates were subjected to mass spectrometry identification methods (Biotyper – MALDI/TOF) and 16S gene sequencing. Results: A total of 105 strains were isolated in this area, bacterial identification by mass spectrometry method (MALDI/TOF) achieved 74% agreement with the conventional identification method (16S), 31% have been unsuccessful in MALDI-TOF and 2% did not obtain identification sequence the 16S. These results show that Biotyper can be a very useful tool in the identification of bacteria isolated from environmental samples, since it has a better value for money (cheap and simple sample preparation and MALDI plates are reusable). Furthermore, this technique is more rentable because it saves time and has a high performance (the mass spectra are compared to the database and it takes less than 2 minutes per sample).

Keywords: copper mining area, bioremediation, microorganisms, identification, MALDI/TOF, RNA 16S

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409 Virtual Screening and in Silico Toxicity Property Prediction of Compounds against Mycobacterium tuberculosis Lipoate Protein Ligase B (LipB)

Authors: Junie B. Billones, Maria Constancia O. Carrillo, Voltaire G. Organo, Stephani Joy Y. Macalino, Inno A. Emnacen, Jamie Bernadette A. Sy

Abstract:

The drug discovery and development process is generally known to be a very lengthy and labor-intensive process. Therefore, in order to be able to deliver prompt and effective responses to cure certain diseases, there is an urgent need to reduce the time and resources needed to design, develop, and optimize potential drugs. Computer-aided drug design (CADD) is able to alleviate this issue by applying computational power in order to streamline the whole drug discovery process, starting from target identification to lead optimization. This drug design approach can be predominantly applied to diseases that cause major public health concerns, such as tuberculosis. Hitherto, there has been no concrete cure for this disease, especially with the continuing emergence of drug resistant strains. In this study, CADD is employed for tuberculosis by first identifying a key enzyme in the mycobacterium’s metabolic pathway that would make a good drug target. One such potential target is the lipoate protein ligase B enzyme (LipB), which is a key enzyme in the M. tuberculosis metabolic pathway involved in the biosynthesis of the lipoic acid cofactor. Its expression is considerably up-regulated in patients with multi-drug resistant tuberculosis (MDR-TB) and it has no known back-up mechanism that can take over its function when inhibited, making it an extremely attractive target. Using cutting-edge computational methods, compounds from AnalytiCon Discovery Natural Derivatives database were screened and docked against the LipB enzyme in order to rank them based on their binding affinities. Compounds which have better binding affinities than LipB’s known inhibitor, decanoic acid, were subjected to in silico toxicity evaluation using the ADMET and TOPKAT protocols. Out of the 31,692 compounds in the database, 112 of these showed better binding energies than decanoic acid. Furthermore, 12 out of the 112 compounds showed highly promising ADMET and TOPKAT properties. Future studies involving in vitro or in vivo bioassays may be done to further confirm the therapeutic efficacy of these 12 compounds, which eventually may then lead to a novel class of anti-tuberculosis drugs.

Keywords: pharmacophore, molecular docking, lipoate protein ligase B (LipB), ADMET, TOPKAT

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