Search results for: gene expression profiles

Authors: Owonikoko Abayomi Dele

Abstract:

Epilepsy is an unresolved disease that needs urgent attention. It is a brain disorder that affects over sixty-five (65) million people around the globe. Despite the availability of commercial anti-epileptic drugs, the war against this unmet condition is yet to be resolved. Most epilepsy patients are resistant to available anti-epileptic medications thus the need for affordable novel therapy against epilepsy is a necessity. Numerous phytochemicals have been reported for their potency, efficacy and safety as therapeutic agents against many diseases. This study investigated 99 isolated phytochemicals from Chasmanthera dependens and Carissa edulis against carbonic anhydrase (ii) drug target. The absorption, distribution, metabolism, excretion and toxicity (ADMET) of the isolated compounds were examined using admet SAR-2 web server while Swiss ADME was used to analyze the oral bioavailability, drug-likeness and lead-likeness properties of the selected leads. PASS web server was used to predict the biological activities of selected leads while other important physicochemical properties and interactions of the selected leads with the active site of the target after successful molecular docking simulation with the pyrx virtual screening tool were also examined. The results of these study identified seven lead compounds; C49- alpha-carissanol (-7.6 kcal/mol), C13- Catechin (-7.4 kcal/mol), C45- Salicin (-7.4 kcal/mol), C6- Bisnorargemonine (-7.3 kcal/mol), C36- Pallidine (-7.1 kcal/mol), S4- Lacosamide (-7.1 kcal/mol), and S7- Acetazolamide (-6.4 kcal/mol) for CA II (3HS4 receptor). These leads compounds are probable inhibitors of this drug target due to the observed good binding affinities and favourable interactions with the active site of the drug target, excellent ADMET profiles, PASS Properties, drug-likeness, lead-likeness and oral bioavailability properties. The identified leads have better binding energies as compared to the binding energies of the two standards. Thus, seven identified lead compounds can be developed further towards the development of new anti-epileptic medications.

Keywords: drug-likeness, phytochemicals, carbonic anhydrases, metalloeazymes, active site, ADMET

Procedia PDF Downloads 58

Authors: Chengcao Sun, Cuili Yang, Shujun Li, Ruilin Xue, Yongyong Xi, Liang Wang, Dejia Li

Abstract:

Backgrounds: Inflammation is widely distributed in patients with Duchenne muscular dystrophy (DMD), and ultimately leads to progressive deterioration of muscle function with the co-effects of chronic muscle damage, oxidative stress, and reduced oxidative capacity. NF-E2-related factor 2 (Nrf2) plays a critical role in defending against inflammation in different tissues via activation of phase II enzymes, heme oxygenase-1 (HO-1). However, whether Nrf2/HO-1 pathway can attenuate muscle inflammation on DMD remains unknown. The purpose of this study was to determine the anti-inflammatory effects of Sulforaphane (SFN) on DMD. Methods: 4-week-old male mdx mice were treated with SFN by gavage (2 mg/kg body weight per day) for 4 weeks. Gastrocnemius, tibial anterior and triceps brachii muscles were collected for related analysis. Immune cell infiltration in skeletal muscles was analyzed by H&E staining and immuno-histochemistry. Moreover, the expressions of inflammatory cytokines,pro-inflammatory cytokines and Nrf2/HO-1 pathway were detected by western blot, qRT-PCR, immunohistochemistry and immunofluorescence assays. Results: Our results demonstrated that SFN treatment increased the expression of muscle phase II enzymes HO-1 in Nrf2 dependent manner. Inflammation in mdx skeletal muscles was reduced by SFN treatment as indicated by decreased immune cell infiltration and lower expressions of the inflammatory cytokines CD45, pro-inflammatory cytokines tumour necrosis factor-α and interleukin-6 in the skeletal muscles of mdx mice. Conclusions: Collectively, these results show that SFN can ameliorate muscle inflammation in mdx mice by Nrf2/HO-1 pathway, which indicates Nrf2/HO-1 pathway may represent a new therapeutic target for DMD.

Keywords: sulforaphane, Nrf2, HO-1, inflammation

Procedia PDF Downloads 336

Authors: Chiara Della Rocca, Gavino Piras, Andrea Berti, Alessandro Mameli

Abstract:

DNA identification of human skeletal remains plays a valuable role in the forensic field, especially in missing persons and mass disaster investigations. Hard tissues, such as bones and teeth, represent a very common kind of samples analyzed in forensic laboratories because they are often the only biological materials remaining. However, the major limitation of using these compact samples relies on the extremely time–consuming and labor–intensive treatment of grinding them into powder before proceeding with the conventional DNA purification and extraction step. In this context, a DNA extraction assay called the TBone Ex kit (DNA Chip Research Inc.) was developed to digest bone chips without powdering. Here, we simultaneously analyzed bone and tooth samples that arrived at our police laboratory and belonged to 15 different forensic casework that occurred in Sardinia (Italy). A total of 27 samples were recovered from different scenarios and were exposed to extreme environmental factors, including sunlight, seawater, soil, fauna, vegetation, and high temperature and humidity. The TBone Ex kit was used prior to the EZ2 DNA extraction kit on the EZ2 Connect Fx instrument (Qiagen), and high-quality autosomal and Y-chromosome STRs profiles were obtained for the 80% of the caseworks in an extremely short time frame. This study provides additional support for the use of the TBone Ex kit for digesting bone fragments/whole teeth as an effective alternative to pulverization protocols. We empirically demonstrated the effectiveness of the kit in processing multiple bone samples simultaneously, largely simplifying the DNA extraction procedure and the good yield of recovered DNA for downstream genetic typing in highly compromised forensic real specimens. In conclusion, this study turns out to be extremely useful for forensic laboratories, to which the various actors of the criminal justice system – such as potential jury members, judges, defense attorneys, and prosecutors – required immediate feedback.

Keywords: DNA, skeletal remains, bones, tbone ex kit, extreme conditions

Procedia PDF Downloads 50

Authors: Navneet Kaur, S. D. Tiwari

Abstract:

Magnetic nanoparticles possess fascinating properties which make their behavior unique in comparison to corresponding bulk materials. Superparamagnetism is one such interesting phenomenon exhibited only by small particles of magnetic materials. In this state, the thermal energy of particles become more than their magnetic anisotropy energy, and so particle magnetic moment vectors fluctuate between states of minimum energy. This situation is similar to paramagnetism of non-interacting ions and termed as superparamagnetism. The magnetization of such systems has been described by Langevin function. But, the estimated fit parameters, in this case, are found to be unphysical. It is due to non-consideration of particle size distribution. In this work, analysis of magnetization data on NiO nanoparticles is presented considering the effect of particle size distribution. Nanoparticles of NiO of two different sizes are prepared by heating freshly synthesized Ni(OH)₂ at different temperatures. Room temperature X-ray diffraction patterns confirm the formation of single phase of NiO. The diffraction lines are seen to be quite broad indicating the nanocrystalline nature of the samples. The average crystallite size are estimated to be about 6 and 8 nm. The samples are also characterized by transmission electron microscope. Magnetization of both sample is measured as function of temperature and applied magnetic field. Zero field cooled and field cooled magnetization are measured as a function of temperature to determine the bifurcation temperature. The magnetization is also measured at several temperatures in superparamagnetic region. The data are fitted to an appropriate expression considering a distribution in particle size following a least square fit procedure. The computer codes are written in PYTHON. The presented analysis is found to be very useful for estimating the particle size distribution present in the samples. The estimated distributions are compared with those determined from transmission electron micrographs.

Keywords: anisotropy, magnetization, nanoparticles, superparamagnetism

Procedia PDF Downloads 143

Authors: Tanakrit Doltanakarn

Abstract:

Introduction: These days, cannabis is being accepted in many countries due to the fact that cannabis could be use in medical. The medical cannabis is used to treat and reduce the pain many diseases. For example, neuropathic pain, Parkinson, autism disorders, cancer pain reduce the adverse effect of chemotherapy, diabetes, and migraine. Active ingredients in cannabis that modulate patients' perceptions of their conditions include Δ9‐tetrahydrocannabinol (THC), cannabidiol (CBD), flavonoids, and terpenes. However, there is an adverse effect of cannabis, cardiovascular effects, psychosis, schizophrenia, mood disorder, and cognitive alternation. These effects are from the THC and CBD ingredients in the cannabis. The metabolize processes of delta-9 THC to 11-OH-delta 9 -THC (inactive form), THC were cause of adverse effects. Interestingly, the distributions of CYP2C9 gene (CYP2C9*2 and CYP2C9*3, poor metabolizer) that might affect incidences of adverse effects in patients who treated with medical cannabis. Objective: The aim of this study we want to investigate the association between genetic polymorphism of CYP2C9 frequency and Thai patients who treated with medical cannabis. Materials and Methods:We recruited sixty-five unrelated Thai patients from the College of Pharmacy, Rangsit University. DNA were extracted using Genomic DNA Mini Kit. Genotyping of CYP2C9*2 (430C>T, rs1799853) and CYP2C9*3 (1075A>C, rs1057910) were genotyped by the TaqMan Real-time PCR assay. Results: Among these 31 medicals cannabis-induced ADRs patients, they were diagnosed with 22 (33.85%) tachycardia and 3 (4.62%) arrhythmia. There were 34 (52.31%) medical cannabis-tolerant controls who were included in this study.40 (61.53%) Thai patients were female, and 25 (38.46%) were male, with median age of 57 (range 27 – 87) years. In this study, we found none of the medical cannabis-induced ADRs carried CYP2C9*2 variant along with medical cannabis-tolerant control group. CYP2C9*3 variant (intermediate metabolizer, IM) was found just only one of thirty-one (3.23%) in the medical cannabis-induced ADRs and two of thirty-fourth (5.88%) in the tolerant controls. Conclusions: Thus, the distribution of CYP2C9 alleles offer a comprehensive view of pharmacogenomics marker in Thai population that could be used as a reference for worldwide to investigate the pharmacogenomics application.

Keywords: medical cannabis, adverse effect, CYP2C9, thai patients

Procedia PDF Downloads 101

Authors: Rose Lastovicka, Bernard Guy, Diana Burton

Abstract:

Greek myth and art acted as tools to think with, and a lens through which to explore complex topics as a form of social media. In particular, coins were a form of propaganda to communicate the wealth and power of the city-states they originated from as they circulated from person to person. From this, how can the application of 3D printing technologies explore the infusion of ancient forms with contemporary commentaries to promote discussion? The digital reconstruction of artifacts is a topic that has been researched by various groups all over the globe. Yet, the exploration of Greek myth through artifacts infused with contemporary issues is currently unexplored in this medium. Using the Stratasys J750 3D printer - a multi-material, full-colour 3D printer - a series of coins inspired by ancient Greek currency and myth was created to present commentaries on the adversities surrounding individuals in the LGBT+ community. Using the J750 as the medium for expression allows for complete control and precision of the models to create complex high-resolution iconography. The coins are printed with a hard, translucent material with coloured 3D visuals embedded into the coin to then be viewed in close contact by the audience. These coins as commentaries present an avenue for wider understanding by drawing perspectives not only from sources concerned with the contemporary LGBT+ community but also from sources exploring ancient homosexuality and the perception and regulation of it in antiquity. By displaying what are usually points of contention between anti- and pro-LGBT+ parties, this visual medium opens up a discussion to both parties, suggesting heritage can play a vital interpretative role in the contemporary world.

Keywords: 3D printing, design, Greek mythology, LGBT+ community

Procedia PDF Downloads 118

Authors: Anahita Suri

Abstract:

Myriad cultures form the warp and weft of the fabric of this world. The last century saw mass migration of people across geographical boundaries, owing to industrialization and globalization. These people took with them their cultures, costumes, traditions, and folklore, which mingled with the local cultures to create something new and place it in a different context to make it contemporary. With the surge in population and growth of the fashion industry, there has been an increasing demand for innovative and individual fashion, from street markets to luxury brands. Exhausted by local influences, designers take inspiration from the so called ‘low’ culture and create artistic products, place it in a different context, and the end-product is categorized as ‘high’ culture. It is challenging as to why a design/culture is ‘high’ or ‘low’. Who decides which works, practices, activities, etc., are ‘high’ and which are ‘low’? The justification for this distinction is often found not in the design itself but the context attached to it. Also, the concept of high/ low is relative to time- what is ‘high’ today can be ‘low’ tomorrow and ‘high’ again the day after. This raises certain concerns. Firstly, it is sad that a culture which offers inspiration is looked down upon as ‘low’ culture. Secondly, it is ironic because the so designated ‘high’ culture is a manipulation of the truth from the authentic ‘low’ culture, which is capable of true expression. When you borrow from a different culture, you pretend to be authentic because you actually are not. Finally, it is important to be aware of crossing cultural boundaries and the context attached to a design/product so as to use it a responsible way that communicates the design without offending anyone. Is it ok for a person’s cultural identity to become another person’s fashion accessory? This essay explores the complex, multi-layered subject of fashion appropriation and aims to provoke debate over cultural ‘borrowing’ and create awareness that commodification of cultural symbols and iconography in fashion is inappropriate and offensive and not the same as ‘celebrating cultural differences’.

Keywords: context, culture, fashion appropriation, inoffensive, responsible

Procedia PDF Downloads 127

Authors: V. Batishchev, V. Getman

Abstract:

We study the self-similar fluid flows in the Marangoni layers with the axial symmetry. Such flows are induced by the radial gradients of the temperatures whose distributions along the free boundary obey some power law. The self-similar solutions describe thermo-capillar flows both in the thin layers and in the case of infinite thickness. We consider both positive and negative temperature gradients. In the former case the cooling of free boundary nearby the axis of symmetry gives rise to the rotation of fluid. The rotating flow concentrates itself inside the Marangoni layer while outside of it the fluid does not revolve. In the latter case we observe no rotating flows at all. In the layers of infinite thickness the separation of the rotating flow creates two zones where the flows are directed oppositely. Both the longitudinal velocity and the temperature have exactly one critical point inside the boundary layer. It is worth to note that the profiles are monotonic in the case of non-swirling flows. We describe the flow outside the boundary layer with the use of self-similar solution of the Euler equations. This flow is slow and non-swirling. The introducing of an outer flow gives rise to the branching of swirling flows from the non-swirling ones. There is such the critical velocity of the outer flow that a non-swirling flow exists for supercritical velocities and cannot be extended to the sub-critical velocities. For the positive temperature gradients there are two non-swirling flows. For the negative temperature gradients the non-swirling flow is unique. We determine the critical velocity of the outer flow for which the branching of the swirling flows happens. In the case of a thin layer confined within free boundaries we show that the cooling of the free boundaries near the axis of symmetry leads to the separating of the layer and creates two sub-layers with opposite rotations inside. This makes sharp contrast with the case of infinite thickness. We show that such rotation arises provided the thickness of the layer exceed some critical value. In the case of a thin layer confined within free and rigid boundaries we construct the branching equation and the asymptotic approximation for the secondary swirling flows near the bifurcation point. It turns out that the bifurcation gives rise to one pair of the secondary swirling flows with different directions of swirl.

Keywords: free surface, rotation, fluid flow, bifurcation, boundary layer, Marangoni layer

Procedia PDF Downloads 345

Authors: Shadia Al-Bahlani, Khadija Al-Bulushi, Zuweina Al-Hadidi, Buthaina Al-Dhahl, Nadia Al-Abri

Abstract:

Breast cancer is the most common cancer in women worldwide. Triple-Negative Breast Cancer (TNBC) is an aggressive type of breast cancer, which is defined by the absence of Estrogen (ER), Progesterone (PR) and human epidermal growth factor (Her-2) receptors. The calpain system plays an important role in many cellular processes including apoptosis, necrosis, cell signaling and proliferation. However, the role of calpain in cisplatin (CDDP)-induced apoptosis in TNBC cells is not fully understood. Here, TNBC (MDA-MB231) cells were treated with different concentration of CDDP (0, 20 & 40 µM) and calpain activation and apoptosis were measured by western blot and Hoechst Stain respectively. In addition, calpain modulation by either activation and/or inhibition and its effect on CDDP-induced apoptosis were assessed by the same above approaches. Our findings showed that CDDP induced endoplasmic reticulum stress and thus Calcium release and subsequently activate calpain α-fodrin cleavage indicated by the increase in GRP78 and Calmodulin protein expression and respectively in MDA-MB231 cells. It also induced apoptosis as measured by Hoechst stain and caspase-12 cleavage. Calpain activation by both Cyclopiazonic acid and Thapsigargin showed similar effect and enhanced the sensitivity of these cells to CDDP treatment. On the other hand, calpain inhibition by either specific siRNA and/or exogenous inhibitor (Calpeptin) had an adverse effect where it attenuated calpain activation and thus CDDP- induced apoptosis in these cells. Altogether, these findings suggested that calpain activation play an essential role in sensitizing the TNBC cells to CDDP-induced apoptosis. This might lead to the discovery of novel treatment to over this aggressive type of breast cancer.

Keywords: calpain, cisplatin, apoptosis, breast cancer

Procedia PDF Downloads 345

Authors: Lahcen Daoudi, Soukaina Elidrissi, Nathalie Fagel

Abstract:

The calcareous accumulations in the surface formations of the soil, are a very widespread phenomenon in the arid and semi-arid regions. Many aspects of physical and chemical evolution of these soils were debated for more than one century. The last two decades have witnessed a remarkable interest in the study of the calcrete. In Morocco, as in most Mediterranean countries, soils with carbonate accumulation cover large areas of the territory. The isohumic subtropical soils and red Mediterranean soils include always a horizon of calcrete accumulation. In the lowland of Tensift Al Haouz located in the central part of Morocco, the arable lands are underlain by indurate pedogenic calcrete of various thicknesses; this constitutes a serious handicap for agricultural development in the region. Our aims in this study is to analyze the characteristics of the crusts developed in this area in order to identify the various facies, their geographic distribution and the factors that played a significant role in the differentiation of these calcareous accumulations. The characterizations were based on various techniques including field observations, X-ray diffraction analysis (XRD) for both raw materials and clay fractions, SEM analysis, Calcimetry and Loss On Ignition (LOI). The analysis of encrusting calcrete in a rich and varied observation field as the region of Tensift Al Haouz enabled us to specify the important types of accumulations: diffuse, nodular and massive encrusting. The shape of encrusting as well as their consistency and hardness is clearly related to the contents of CaCO3 of the profiles. Among these facies, the hardpan which results from a complex succession of processes is certainly the most morphologically advanced form of encrusting. The vertical and lateral distribution of these forms in the Tensift Al Haouz area indicates that they do not appear randomly but seem related to well defined environmental conditions. The differentiation and evolution of encrusting is under the influence of two major factors: 1) the availability of carbonate rich solution which is controlled by the topography, the nature and texture of underlying host rock and the detrital processes; 2) the climate which is responsible for the evaporation and crystallization of carbonate.

Keywords: soil calcrete, characterization, morphology, Tensift Al Haouz, Morocco

Procedia PDF Downloads 401

Authors: Azam Zare

Abstract:

Separation flow control for performance enhancement over airfoils at high incidence angle has become an increasingly important topic. This work details the characteristics of an efficient feedback control of the turbulent subsonic flow over NACA23012 airfoil using forced reduced‐order model based on the proper orthogonal decomposition/Galerkin projection and perturbation method on the compressible Reynolds Averaged Navier‐Stokes equations. The forced reduced‐order model is used in the optimal control of the turbulent separated flow over a NACA23012 airfoil at Mach number of 0.2, Reynolds number of 5×106, and high incidence angle of 24° using blowing/suction controlling jets. The Spallart-Almaras turbulence model is implemented for high Reynolds number calculations. The main shortcoming of the POD/Galerkin projection on flow equations for controlling purposes is that the blowing/suction controlling jet velocity does not show up explicitly in the resulting reduced order model. Combining perturbation method and POD/Galerkin projection on flow equations introduce a forced reduced‐order model that can predict the time-varying influence of the blowing/suction controlling jet velocity. An optimal control theory based on forced reduced‐order system is used to design a control law for a nonlinear reduced‐order model, which attempts to minimize the vorticity content in the turbulent flow field over NACA23012 airfoil. Numerical simulations were performed to help understand the behavior of the controlled suction jet at 12% to 18% chord from leading edge and a pair of blowing/suction jets at 15% to 18% and 24% to 30% chord from leading edge, respectively. Analysis of streamline profiles indicates that the blowing/suction jets are efficient in removing separation bubbles and increasing the lift coefficient up to 22%, while the perturbation method can predict the flow field in an accurate Manner.

Keywords: flow control, POD, Galerkin projection, separation

Procedia PDF Downloads 155

Authors: Minoru Moriguchi

Abstract:

This research introduces the concept of a monolingual Japanese dictionary for non-native speakers of Japanese, whose temporal title is Dictionary of Contemporary Japanese for Advanced Learners (DCJAL). As the language market is very small compared with English, a monolingual Japanese dictionary for non-native speakers, containing sufficient entries, has not been published yet. In such a dictionary environment, Japanese-language learners are using bilingual dictionaries or monolingual Japanese dictionaries for Japanese people. This research started in 2017, as a project team which consists of four Japanese and two non-native speakers, all of whom are linguists of the Japanese language. The team has been trying to propose the concept of a monolingual dictionary for non-native speakers of Japanese and to provide the entry list, the definition samples, the list of defining vocabulary, and the writing manual. As the result of seven-year research, DCJAL has come to have 28,060 head words, 539 entry examples, 4,598-word defining vocabulary, and the writing manual. First, the number of the entry was determined as about 30,000, based on an experimental method using existing six dictionaries. To make the entry list satisfying this number, words suitable for DCJAL were extracted from the Tsukuba corpus of the Japanese language, and later the entry list was adjusted according to the experience as Japanese instructor. Among the head words of the entry list, 539 words were selected and added with lexicographical information such as proficiency level, pronunciation, writing system (hiragana, katakana, kanji, or alphabet), definition, example sentences, idiomatic expression, synonyms, antonyms, grammatical information, sociolinguistic information, and etymology. While writing the definition of the above 539 words, the list of the defining vocabulary was constructed, based on frequent vocabulary used in a Japanese monolingual dictionary. Although the concept of DCJAL has been almost perfected, it may need some more adjustment, and the research is continued.

Keywords: monolingual dictionary, the Japanese language, non-native speaker of Japanese, defining vocabulary

Procedia PDF Downloads 43

Authors: Wong Yat Cheung Maggie

Abstract:

Occupational Safety and Health Ordinance (OS&HO) (Chap 509) was enacted in 1997, OHS in HK should be growing and maturing, with a holistic approach to occupational health and safety in the workplace including physical, mental, social and spiritual well-being. The question is “How effective are OHSPs in meeting the current needs of HK health workers?” This study was designed to explore the issue for the first time, to empirically analyse the views of those who work in the system. The study employed a mixed method approach to collect various data from Occupational Health Service Providers (OHSPs), Occupational Health Service Consumers (OHSC): Registered nurses working in the hospital setting. This study was designed in two phases and two stages. Phase I Stage I was a paper survey to collect the data on OHSP. Then Phase I Stage II was a follow-up interview. Phase II Stage I was a paper survey to collect the data on OHSC. Then Phase II Stage II was a follow-up focus group study on OHSC for further clarification of the Phase II and Stage I result. The Phase I result reflects HK OHSPs point of view and their experience in the existing OHS practice in the local hospitals. It reflects various styles of reporting systems, staff profiles background and resource in providing OHS in HK hospitals. However, the basic OHS concern is similar between hospitals. In general, the OHS policies and procedures are available on site even though they may have different foci. The Phase II result is reflecting the HKs OHSCs echoes the OHSP feedback at providing of OHS, OHS concern and related policies and procedure are available on site. However, the most significant feedback from the OHSC at Phase II Stage II shows, nurses experienced various OHS concern most commonly work stress, workplace harassment and back strain without formal or official report to the related parties. The lack of reporting was due to the management handling attitude, stakeholders’ compliance and term of definition still have room to be improved even the related policies and procedures are available on site.

Keywords: occupational health service, registered nurse, Hong Kong hospital, mixed method

Procedia PDF Downloads 335

Authors: Abdul Soofi, Katherine Wolf, Egon Ranghini, Gregory Dressler

Abstract:

Obesity and its associated complications, such as insulin resistance and non-alcoholic fatty liver disease, are reaching epidemic proportions. In mice, the TGF-β superfamily is implicated in the regulation of white and brown adipose tissues differentiation. The Kielin/Chordin-like Protein (KCP) is a secreted regulator of the TGF-β superfamily pathways that can inhibit both TGF-β and Activin signals while enhancing the Bone Morphogenetic protein (BMP) signaling. However, the effects of KCP on metabolism and obesity have not been studied in animal models. Thus, we examined the effects of KCP loss or gain of function in mice that were maintained on either a regular or a high fat diet. Loss of KCP sensitized mice to obesity and associated complications such as hepatic steatosis and glucose intolerance. In contrast, transgenic mice that expressed KCP in the kidney, liver and adipose tissues were resistant to developing high fat diet induced obesity and had significantly reduced white adipose tissue. KCP over-expression was able to shift the pattern of Smad signaling in vivo, to increase the levels of P-Smad1 and decrease P-Smad3, resulting in resistance to high fat diet induced hepatic steatosis and glucose intolerance. In aging mice, loss of KCP promoted liver pathology even when mice were fed a normal diet. The data demonstrate that shifting the TGF-β superfamily signaling with a secreted inhibitor or enhancer can alter the physiology of adipose tissue to reduce obesity and can inhibit the initiation and progression of hepatic steatosis to significantly reduce the effects of high fat diet induced metabolic disease.

Keywords: adipose tissue, KCP, obesity, TGF-β, BMP, hepatic steatosis, metabolic syndrome

Procedia PDF Downloads 353

Authors: Worku Dejene Bekele, András Marczika Csilla Sörös

Abstract:

Silver nanoparticles are silver metal with a size of 1-100nm. The most common source of silver nanoparticles is inorganic salts. Nanoparticles can be ingested through our foods and constitute nanoparticles and silver ions, whether as an additive or by migrants and, in some cases, as a pollutant. Silver nanoparticles are the most widely applicable engineered nanomaterials, especially for antimicrobial function. Ag nanoparticles give different advantages in the case of food safety, quality, and overall acceptability; however, they affect the health of humans and animals, putting them at risk of health problems and environmental pollution. Silver nanoparticles have been used widely in food packaging technologies, especially in water treatments, meat and meat products, fruit, and many other food products. This is for bio-preservation from food products. The primary goal of this review is to determine the safety and health impact of Ag nanoparticles application in food packaging and analysis of the human organs more affected by this preservative technology, to assess the implications of a nanoparticle on food safety, to determine the effects of nanoparticles on consumers health and to determine the impact of nanotechnology on product acceptability. But currently, much research has demonstrated that there is cause to believe that silver nanoparticles may have toxicological effects on biological organs and systems. The silver nanoparticles affect DNA expression, gastrointestinal barriers, lungs, and other breathing organs illness. Silver particles and molecules are very toxic. During its application in food packaging, food industries used the thinnest particle. This particle can potentially affect the gastrointestinal tracts-it suffers from mucus production, DNA, lungs, and other breezing organs. This review is targeted to demonstrate the knowledge gap that industrials use in the application of silver nanoparticles in food packaging and preservation and its health effects on the consumer.

Keywords: food preservatives, health impact, nanoparticle, silver nanoparticle

Procedia PDF Downloads 71

Authors: John Tate

Abstract:

In 2004, the French Parliament banned the “hijab” in public schools, and in 2010 it prohibited the “burqa” and “niqab” in “public places.” The result was a “secular” outcome involving the removal of these garments, often identified with Islamic religious and cultural practice, from the French public sphere. Yet in 2016, the French local council bans on the “burqini” were overruled by France’s highest administrative court, the Conseil d’État, allowing for their retention in the public sphere. Unlike the burqa and hijab bans, the burqini bans produced significant divisions at the highest echelons of the French political class, with the Prime Minister, Manuel Valls, and the President, François Hollande, finding themselves at odds on the issue. This article seeks to achieve four aims. It seeks to (a) explain the contrary outcomes between key French state institutions, such as the Conseil d’État and the French Parliament, concerning the hijab and burqa bans, and the Conseil d’État and French local councils, concerning the burqini bans; (b) to do so by identifying two qualitatively distinct, and at times incompatible, conceptions of laïcité, present within official French public discourse, and applied by these French state institutions to underwrite these respective outcomes; (c) explain why, given these contrary conceptions of laïcité, and these contrary outcomes, the widespread identification of laïcité with “secularism” is both misleading and inaccurate; and (d) provide an explanation why senior members of the French political class were divided on the burqini bans when they were not divided on the nation-wide prohibitions of the hijab in public schools and the burqa in public places. In regard to this last question, the article seeks to ask why the Burqini was “different”?

Keywords: liberalism, republicanism, laïcité, citizenship

Procedia PDF Downloads 149

Authors: Yangwon Hyun

Abstract:

Nowadays, business settings are becoming increasingly multilingual and multicultural under the conditions of globalization. When people from different linguistic and cultural background interact, their divergent cultural conceptualizations may result in communicative problems. Even though intercultural business communication between Thais and South Koreans has become increasingly significant along with the development of international trade relations, there have been very few attempts to conduct research on interactional problems in that context. Based on a Cultural Linguistics perspective, this study analyzes Thai cultural conceptualizations that underly interactional problems with Koreans at Korean business organizations where were located in Thailand. The data was gathered through in-depth interviews with ten Korean and ten Thai employees as well as through six months of field observations. The results indicate that there are three cultural pragmatic schemas that underly problems of interaction between the two groups; (1) the schema of kreng-jai as a communicative strategy of withholding one’s intentions in order to not hurt another’s emotion, (2) the schema of mai-pen-rai as a linguistic strategy of inducing interlocutors to not be concerned about substantial matters so as to avoid confrontations, and (3) the schema of jai-yen as a linguistic expression used for persuading interlocutors to maintain a calm state of mind in order to avoid conflicts. The linguistic behaviors of Thai employees in relation to these three cultural schemas had led to misunderstandings and interpersonal conflicts with Korean supervisors and co-workers. These interactional problems were caused not only because Koreans were unfamiliar with Thai cultural conceptualizations, but also because these conceptualizations were incompatible with goal-oriented Korean business discourse. The findings of this study can enhance mutual understandings between Thai and Korean employees. Thus, it contributes to the promotion of smooth intercultural communication in the workplace.

Keywords: business context, cultural conceptualizations, cultural linguistics, intercultural communication, thai culture

Procedia PDF Downloads 132

Authors: Jayanth Abishek Subramanian, Ramin Dabirian, Ilias Gavrielatos, Ram Mohan, Ovadia Shoham

Abstract:

Downcomers are important conduits for multiphase flow transfer from offshore platforms to the seabed. Uncertainty in the predictions of the pressure drop of multiphase flow between platforms is often dominated by the uncertainty associated with the prediction of holdup and pressure drop in the downcomer. The objectives of this study are to conduct experimental and theoretical scale-up study of the downcomer. A 4-in. diameter vertical test section was designed and constructed to study two-phase flow in downcomer. The facility is equipped with baffles for flow area restriction, enabling interchangeable annular slot openings between 30% and 61.7%. Also, state-of-the-art instrumentation, the capacitance Wire-Mesh Sensor (WMS) was utilized to acquire the experimental data. A total of 76 experimental data points were acquired, including falling film under 30% and 61.7% annular slot opening for air-water and air-Conosol C200 oil cases as well as gas carry-under for 30% and 61.7% opening utilizing air-Conosol C200 oil. For all experiments, the parameters such as falling film thickness and velocity, entrained liquid holdup in the core, gas void fraction profiles at the cross-sectional area of the liquid column, the void fraction and the gas carry under were measured. The experimental results indicated that the film thickness and film velocity increase as the flow area reduces. Also, the increase in film velocity increases the gas entrainment process. Furthermore, the results confirmed that the increase of gas entrainment for the same liquid flow rate leads to an increase in the gas carry-under. A power comparison method was developed to enable evaluation of the Lopez (2011) model, which was created for full bore downcomer, with the novel scale-up experiment data acquired from the downcomer with the restricted area for flow. Comparison between the experimental data and the model predictions shows a maximum absolute average discrepancy of 22.9% and 21.8% for the falling film thickness and velocity, respectively; and a maximum absolute average discrepancy of 22.2% for fraction of gas carried with the liquid (oil).

Keywords: two phase flow, falling film, downcomer, wire-mesh sensor

Procedia PDF Downloads 167

Authors: V. Kuzhandai Velu, R. Ramesh

Abstract:

Background and objectives: Acute myocardial infarction (AMI) is the most common cause of mortality and morbidity. Over the last decade, microRNAs (miRs) have emerged as a potential marker for detecting AMI. The current study evaluates the kinetics and importance of miRs in the differential diagnosis of ST-segment elevated MI (STEMI) and non-STEMI (NSTEMI) and its correlation to conventional biomarkers and to predict the immediate outcome of AMI for arrhythmias and left ventricular (LV) dysfunction. Materials and Method: A total of 100 AMI patients were recruited for the study. Routine cardiac biomarker and miRNA levels were measured during diagnosis and serially at admission, 6, 12, 24, and 72hrs. The baseline biochemical parameters were analyzed. The expression of miRs was compared between STEMI and NSTEMI at different time intervals. Diagnostic utility of miR-1, miR-133, miR-208, and miR-499 levels were analyzed by using RT-PCR and with various diagnostics statistical tools like ROC, odds ratio, and likelihood ratio. Results: The miR-1, miR-133, and miR-499 showed peak concentration at 6 hours, whereas miR-208 showed high significant differences at all time intervals. miR-133 demonstrated the maximum area under the curve at different time intervals in the differential diagnosis of STEMI and NSTEMI which was followed by miR-499 and miR-208. Evaluation of miRs for predicting arrhythmia and LV dysfunction using admission sample demonstrated that miR-1 (OR = 8.64; LR = 1.76) and miR-208 (OR = 26.25; LR = 5.96) showed maximum odds ratio and likelihood respectively. Conclusion: Circulating miRNA showed a highly significant difference between STEMI and NSTEMI in AMI patients. The peak was much earlier than the conventional biomarkers. miR-133, miR-208, and miR-499 can be used in the differential diagnosis of STEMI and NSTEMI, whereas miR-1 and miR-208 could be used in the prediction of arrhythmia and LV dysfunction, respectively.

Keywords: myocardial infarction, cardiac biomarkers, microRNA, arrhythmia, left ventricular dysfunction

Procedia PDF Downloads 128

Authors: S. Blanchard, D. Beladjine, K. Beddiar

Abstract:

Since 2015, the arrival of BIM in the building sector in France has turned the corporation world upside down. Not only constructive practices have been impacted, but also the uses and the men who have undergone important changes. Thus, the new collaborative mode generated by the BIM and the digital model has challenged the supremacy of some construction actors because the process involves working together taking into account the needs of other contributors. New BIM tools have emerged and actors in the act of building must take ownership of them. It is in this context that under the impetus of a European directive and the French government's encouragement of new missions and job profiles have. Moreover, concurrent engineering requires that each actor can advance at the same time as the others, at the whim of the information that reaches him, and the information he has to transmit. However, in the French legal system around public procurement, things are not planned in this direction. Also, a consequent evolution must take place to adapt to the methodology. The new missions generated by the BIM in France require a good mastery of the tools and the process. Also, to meet the objectives of the BIM approach, it is possible to define a typical job profile around the BIM, adapted to the various sectors concerned. The multitude of job offers using the same terms with very different objectives and the complexity of the proposed missions motivated by our approach. In order to reinforce exchanges with professionals or specialists, we carried out a statistical study to answer this problem. Five topics are discussed around the business area: the BIM in the company, the function (business), software used and BIM missions practiced (39 items). About 1400 professionals were interviewed. These people work in companies (micro businesses, SMEs, and Groups) of construction, engineering offices or, architectural agencies. 77% of respondents have the status of employees. All participants are graduated in their trade, the majority having level 1. Most people have less than a year of experience in BIM, but some have 10 years. The results of our survey help to understand why it is not possible to define a single type of BIM Manager. Indeed, the specificities of the companies are so numerous and complex and the missions so varied, that there is not a single model for a function. On the other hand, it was possible to define 3 main professions around the BIM (Manager, Coordinator and Modeler) and 3 main missions for the BIM Manager (deployment of the method, assistance to project management and management of a project).

Keywords: BIM manager, BIM modeler, BIM coordinator, project management

Procedia PDF Downloads 164

Authors: Temesgen Minase Woldegebriel

Abstract:

This study undertakes a comparative analysis of two distinct staples from diverse culinary heritages: Polish traditional bread and Teff Injera. Despite originating from disparate cultural contexts, both these foods hold significant roles in their respective societies, serving as dietary staples rich in cultural symbolism and nutritional value. Our investigation delves into the historical, cultural, and nutritional dimensions of Polish bread and Teff Injera, shedding light on their ingredients, preparation methods, and consumption patterns. Firstly, we explore the rich history and cultural significance embedded within Polish traditional bread, tracing its evolution through centuries of tradition and craftsmanship. From the ubiquitous Polish Rye bread to the intricate regional variations, we unravel the socio-cultural narratives intertwined with each loaf, reflecting Polish identity and culinary heritage. In contrast, our analysis extends to Teff Injera, a staple of Ethiopian and Eritrean cuisine known for its spongy texture and tangy flavor. We delve into the ancient origins of Teff cultivation, highlighting its pivotal role in Ethiopian culture and its symbolic significance in communal dining practices, such as the traditional Ethiopian coffee ceremony. Furthermore, we undertake a comparative examination of the nutritional profiles of Polish bread and Teff Injera, assessing their respective contributions to dietary health and well-being. Through comprehensive nutritional analysis, we elucidate the unique attributes of each staple, considering factors such as gluten content, fiber composition, and micronutrient density. Moreover, our study investigates the contemporary relevance of these traditional staples in the context of shifting dietary preferences and global culinary trends. We analyze consumer perceptions and market dynamics surrounding Polish bread and Teff Injera, discerning patterns of consumption and avenues for innovation in a rapidly evolving food landscape. In conclusion, our comparative analysis illuminates the multifaceted dimensions of Polish traditional bread and Teff Injera, transcending mere culinary discourse to encompass broader themes of cultural heritage, nutrition, and gastronomic diversity.

Keywords: bread, culinary, injera, teff

Procedia PDF Downloads 18

Authors: Jana Stepanovska, Roman Matejka, Jozef Rosina, Marta Vandrovcova, Lucie Bacakova

Abstract:

The aim of this study was to develop a system for mechanical and also electrical stimulation controlling in vitro osteogenesis under conditions more similar to the in vivo bone microenvironment than traditional static cultivation, which would achieve good adhesion, growth and other specific behaviors of osteogenic cells in cultures. An engineered culture system for mechanical stimulation of the mesenchymal stem cells on the charged surface was designed. The bioreactor allows efficient mechanical loading inducing an electrical response and perfusion of the culture chamber with seeded cells. The mesenchymal stem cells were seeded to specific charged materials, like polarized hydroxyapatite (Hap) or other materials with piezoelectric and ferroelectric features, to create electrical potentials for stimulating of the cells. The material of the matrix was TiNb alloy designed for these purposes, and it was covered by BaTiO3 film, like a kind of piezoelectric material. The process of mechanical stimulation inducing electrical response is controlled by measuring electrical potential in the chamber. It was performed a series of experiments, where the cells were seeded, perfused and stimulated up to 48 hours under different conditions, especially pressure and perfusion. The analysis of the proteins expression was done, which demonstrated the effective mechanical and electrical stimulation. The experiments demonstrated effective stimulation of the cells in comparison with the static culture. This work was supported by the Ministry of Health, grant No. 15-29153A and the Grant Agency of the Czech Republic grant No. GA15-01558S.

Keywords: charged surface, dynamic cultivation, electrical stimulation, ferroelectric layers, mechanical stimulation, piezoelectric layers

Procedia PDF Downloads 301

Authors: Marzieh Khedmati, Shannon L. Bartelt-Hunt

Abstract:

Vegetative Filter Strips (VFS) are used for controlling the volume of runoff and decreasing contaminant concentrations in runoff before entering water bodies. Many studies have investigated the role of VFS in sediment and nutrient removal, but little is known about their efficiency for the removal of emerging contaminants such as antimicrobial resistance genes (ARGs). Vegetative Filter Strip Modeling System (VFSMOD) was used to simulate the efficiency of VFS in this regard. Several studies demonstrated the ability of VFSMOD to predict reductions in runoff volume and sediment concentration moving through the filters. The objectives of this study were to calibrate the VFSMOD with experimental data and assess the efficiency of the model in simulating the filter behavior in removing ARGs (ermB) and tylosin. The experimental data were obtained from a prior study conducted at the University of Nebraska (UNL) Rogers Memorial Farm. Three treatment factors were tested in the experiments, including manure amendment, narrow grass hedges and rainfall events. Sediment Delivery Ratio (SDR) was defined as the filter efficiency and the related experimental and model values were compared to each other. The VFS Model generally agreed with the experimental results and as a result, the model was used for predicting filter efficiencies when the runoff data are not available. Narrow Grass Hedges (NGH) were shown to be effective in reducing tylosin and ARGs concentration. The simulation showed that the filter efficiency in removing ARGs is different for different soil types and filter lengths. There is an optimum length for the filter strip that produces minimum runoff volume. Based on the model results increasing the length of the filter by 1-meter leads to higher efficiency but widening beyond that decreases the efficiency. The VFSMOD, which was proved to work well in estimation of VFS trapping efficiency, showed confirming results for ARG removal.

Keywords: antimicrobial resistance genes, emerging contaminants, narrow grass hedges, vegetative filter strips, vegetative filter strip modeling system

Procedia PDF Downloads 133

Authors: G. Scremin, I. Y. Suh, S. Doukas

Abstract:

Spectators follow their favorite sports teams for different reasons. While some attend a sporting event simply for its entertainment value, others do so because of the personal sense of achievement and accomplishment their connection with a sports team creates. Moreover, the level of identity spectators feel toward their favorite sports team falls in a broad continuum. Some are mere spectators. For those spectators, their association to a sports team has little impact on their self-image. Others are die-hard fans who are proud of their association with their team and whose connection with that team is an important reflection of who they are. Several motives for sports consumption can be used to explain the level of spectator support in a variety of sports. Those motives can also be used to explain the variance in the identification, attachment, and loyalty spectators feel toward their favorite sports team. Motives for sports consumption can be used to discriminate the degree of identification spectators have with their favorite sports team. In this study, motives for sports consumption was used to discriminate the level of identity spectators feel toward their sports team. It was hypothesized that spectators with a strong level of team identity would report higher rates of interest in player, interest in sports, and interest in team than spectators with a low level of team identity. And spectators with a low level of team identity would report higher rates for entertainment value, bonding with friends or family, and wholesome environment. Football spectators in the United States and England were surveyed about their motives for football consumption and their level of identification with their favorite football team. To assess if the motives of sports fans differed by level of team identity and allegiance to an American or English football team, a Multivariate Analysis of Variance (MANOVA) under the General Linear Model (GLM) procedure found in SPSS was performed. The independent variables were level of team identity and allegiance to an American or English football team, and the dependent variables were the sport fan motives. A tripartite split (low, moderate, high) was used on a composite measure for team identity. Preliminary results show that effect of team identity is statistically significant (p < .001) for at least nine of the 17 motives for sports consumption assessed in this investigation. These results indicate that the motives of spectators with a strong level of team identity differ significantly from spectators with a low level of team identity. Those differences can be used to discriminate the degree of identification spectators have with their favorite sports team. Sports marketers can use these methods and results to develop identity profiles of spectators and create marketing strategies specifically designed to attract those spectators based on their unique motives for consumption and their level of team identification.

Keywords: fan identification, market segmentation of sports fans, motives for sports consumption, team identity

Procedia PDF Downloads 170

Authors: Frazer G. Thompson

Abstract:

The 1990s in America ended with a mass-school-shooting (at least four killed by gunfire excluding the perpetrator(s)) at Columbine High School in Littleton, Colorado. Post-event, many demanded that government and civilian experts develop a ‘profile’ of the potential school shooter in order to identify and preempt likely future acts of violence. This grounded theory research study seeks to explore the validity of the original hypotheses proposed by the Federal Bureau of Investigation (FBI) in 2000, as it relates to the commonality of disclosure by perpetrators of mass-school-shootings, by evaluating fourteen mass-school-shooting events between 2000 and 2019 at locations around the United States. Methods: The strategy of inquiry seeks to investigate case files, public records, witness accounts, and available psychological profiles of the shooter. The research methodology is inclusive of one-on-one interviews with members of the FBI’s Critical Incident Response Group seeking perspective on commonalities between individuals; specifically, disclosure of intent pre-event. Results: The research determined that school shooters do not ‘unfailingly’ notify others of their plans. However, in nine of the fourteen mass-school-shooting events analyzed, the perpetrator did inform the third party of their intent pre-event in some form of written, oral, or electronic communication. In the remaining five instances, the so-called ‘red-flag’ indicators of the potential for an event to occur were profound, and unto themselves, might be interpreted as notification to others of an imminent deadly threat. Conclusion: Data indicates that conclusions drawn in the FBI’s threat assessment perspective published in 2000 are relevant and current. There is evidence that despite potential ‘red-flag’ indicators which may or may not include a variety of other characteristics, perpetrators of mass-school-shooting events are likely to share their intentions with others through some form of direct or indirect communication. More significantly, implications of this research might suggest that society is often informed of potential danger pre-event but lacks any equitable means by which to disseminate, prevent, intervene, or otherwise act in a meaningful way considering said revelation.

Keywords: columbine, FBI profiling, guns, mass shooting, mental health, school violence

Procedia PDF Downloads 120

Authors: M. Nita-Lazar, E. Manea, C. Bumbac, A. Banciu, C. Stoica

Abstract:

The population and economic growth have generated a significant new number of pollutant compounds which have to be degraded before reaching the environment. The wastewater treatment plants (WWTPs) have been the last barrier between the domestic and/or industrial wastewaters and the environment. At present, the conventional WWTPs have very high operational costs, most of them linked to the aeration process (60-65% from total energy costs related to wastewater treatment). In addition, they have had a low efficiency in pollutants removal such as pharmaceutical and other resilient anthropogenic compounds. In our study, we have been focused on new wastewater treatment strategies to enhance the efficiency of pollutants removal and decrease the wastewater treatment operational costs. The usage of mixed microalgae-bacteria granules technology generated high efficiency and low costs by a better harvesting and less expensive aeration. The intertrophic relationships between microalgae and bacteria have been characterized by the structure of the population community to their metabolic relationships. The results, obtained by microscopic studies, showed well-organized and stratified microalgae-bacteria granules where bacteria have been enveloped in the microalgal structures. Moreover, their population community structure has been modulated as well as their nitrification, denitrification processes (analysis based on qPCR genes expression) by the type of the pollutant compounds and amounts. In conclusion, the understanding and modulation of intertrophic relationships between microalgae and bacteria could be an economical and technological viable alternative to the conventional wastewater treatment. Acknowledgements: This research was supported by grant PN-III-P4-ID-PCE-2016-0865 from the Romanian National Authority for Scientific Research and Innovation CNCS/CCCDI-UEFISCDI.

Keywords: activated sludge, bacteria, granules, microalgae

Procedia PDF Downloads 125

Authors: Ahmed M. Kabel, Maaly A. Abd Elmaaboud

Abstract:

Scleroderma is one of the connective tissue disorders characterized by skin and systemic fibrosis. Its pathogenesis involves multiple interrelated processes of autoimmunity, vasculopathy, inflammation, and oxidative stress. This study was a trial to explore the possible ameliorative effects of sodium valproate on an experimental model of skin fibrosis induced by bleomycin. Forty male BALB/c mice were divided into four equal groups as follows: control group; bleomycin group; bleomycin + sodium valproate group, and sodium valproate group. Mice were assessed for their body weight every four days throughout the whole study. Skin tissues were used to evaluate the oxidative stress parameters, transforming growth factor beta 1 (TGF-β1), tumor necrosis factor alpha (TNF-α), interleukin 15, and mammalian target of rapamycin (mTOR). Skin fibrosis was evaluated by measuring dermal thickness and staining the skin tissues with Masson trichrome stain. Also, the skin tissues were immunostained with alpha smooth muscle actin (α-SMA). Administration of sodium valproate to bleomycin-treated mice resulted in the restoration of the body weight with a significant decrease in the dermal thickness, amelioration of oxidative stress, suppression of TGF-β1 and mTOR expression, and significant reduction of the percentage of α-SMA immunostaining and the proinflammatory cytokine levels compared to mice treated with bleomycin alone. In conclusion, sodium valproate has an antifibrotic effect on skin fibrosis which may represent a beneficial therapeutic modality for the management of scleroderma.

Keywords: scleroderma, bleomycin, sodium valproate, skin fibrosis

Procedia PDF Downloads 83

Authors: Kirsten Wilson, Patrick M. Brauer, Sandra Babic, Diana Golubeva, Jessica Van Eyk, Tinya Wang, Avanti Karkhanis, Tim A. Le Fevre, Andy I. Kokaji, Allen C. Eaves, Sharon A. Louis, , Nooshin Tabatabaei-Zavareh

Abstract:

Long-lived plasma cells are rare, non-proliferative B cells generated from antibody-secreting cells (ASCs) following an immune response to protect the host against pathogen re-exposure. Despite their therapeutic potential, the lack of in vitro protocols in the field makes it challenging to use B cells as a cellular therapeutic tool. As a result, there is a need to establish robust and reproducible methods for the generation of B cells. To address this, we have developed a culture system for generating B cells from hematopoietic stem and/or progenitor cells (HSPCs) derived from human umbilical cord blood (CB) or pluripotent stem cells (PSCs). HSPCs isolated from CB were cultured using the StemSpan™ B Cell Generation Kit and produced CD19+ B cells at a frequency of 23.2 ± 1.5% and 59.6 ± 2.3%, with a yield of 91 ± 11 and 196 ± 37 CD19+ cells per input CD34+ cell on culture days 28 and 35, respectively (n = 50 - 59). CD19+IgM+ cells were detected at a frequency of 31.2 ± 2.6% and were produced at a yield of 113 ± 26 cells per input CD34+ cell on culture day 35 (n = 50 - 59). The B cell receptor loci of CB-derived B cells were sequenced to confirm V(D)J gene rearrangement. ELISpot analysis revealed that ASCs were generated at a frequency of 570 ± 57 per 10,000 day 35 cells, with an average IgM+ ASC yield of 16 ± 2 cells per input CD34+ cell (n = 33 - 42). PSC-derived HSPCs were generated using the STEMdiff™ Hematopoietic - EB reagents and differentiated to CD10+CD19+ B cells with a frequency of 4 ± 0.8% after 28 days of culture (n = 37, 1 embryonic and 3 induced pluripotent stem cell lines tested). Subsequent culture of PSC-derived HSPCs increased CD19+ frequency and generated ASCs from 1 - 2 iPSC lines. This method is the first report of a serum- and feeder-free system for the generation of B cells from CB and PSCs, enabling further B lineage-specific research for potential future clinical applications.

Keywords: stem cells, B cells, immunology, hematopoiesis, PSC, differentiation

Procedia PDF Downloads 59

Authors: Marinna Madrid, Nabiha Saklayen, Marinus Huber, Nicolas Vogel, Christos Boutopoulos, Michel Meunier, Eric Mazur

Abstract:

Delivering material into cells is important for a diverse range of biological applications, including gene therapy, cellular engineering and imaging. We present a plasmonic substrate for delivering membrane-impermeable material into cells at high throughput and high efficiency while maintaining cell viability. The substrate fabrication is based on an affordable and fast colloidal self-assembly process. When illuminated with a femtosecond laser, the light interacts with the electrons at the surface of the metal substrate, creating localized surface plasmons that form bubbles via energy dissipation in the surrounding medium. These bubbles come into close contact with the cell membrane to form transient pores and enable entry of membrane-impermeable material via diffusion. We use fluorescence microscopy and flow cytometry to verify delivery of membrane-impermeable material into HeLa CCL-2 cells. We show delivery efficiency and cell viability data for a range of membrane-impermeable cargo, including dyes and biologically relevant material such as siRNA. We estimate the effective pore size by determining delivery efficiency for hard fluorescent spheres with diameters ranging from 20 nm to 2 um. To provide insight to the cell poration mechanism, we relate the poration data to pump-probe measurements of micro- and nano-bubble formation on the plasmonic substrate. Finally, we investigate substrate stability and reusability by using scanning electron microscopy (SEM) to inspect for damage on the substrate after laser treatment. SEM images show no visible damage. Our findings indicate that self-assembled plasmonic substrates are an affordable tool for high-throughput, high-efficiency delivery of material into mammalian cells.

Keywords: femtosecond laser, intracellular delivery, plasmonic, self-assembly

Procedia PDF Downloads 531

Authors: Dong-Gyu Leem, Ji-Sun Shin, Sang Yoon Choi, Kyung-Tae Lee

Abstract:

In this study, we focused on the anti-proliferative effects of panaxydol, a C17 polyacetylenic compound derived from Panax ginseng roots, against various human cancer cells. We treated with panaxydol to various cancer cells and panaxydol treatment was found to significantly inhibit the proliferation of human lung cancer cells (A549) and human pancreatic cancer cells (AsPC-1 and MIA PaCa-2), of which AsPC-1 cells were most sensitive to its treatment. DNA flow cytometric analysis indicated that panaxydol blocked cell cycle progression at the G1 phase in A549 cells, which accompanied by a parallel reduction of protein expression of cyclin-dependent kinase (CDK) 2, CDK4, CDK6, cyclin D1 and cyclin E. CDK inhibitors (CDKIs), such as p21CIP1/WAF1 and p27KIP1, were gradually upregulated after panaxydol treatment at the protein levels. Furthermore, panaxydol induced the activation of p53 in A549 cells. In addition, panaxydol also induced apoptosis of AsPC-1 and MIA PaCa-2 cells, as shown by accumulation of subG1 and apoptotic cell populations. Panaxydol triggered the activation of caspase-3, -8, -9 and the cleavage of poly (ADP-ribose) polymerase (PARP). Reduction of mitochondrial transmembrane potential by panaxydol was determined by staining with dihexyloxacarbocyanine iodide. Furthermore, panaxydol suppressed the levels of anti-apoptotic proteins, XIAP and Bcl-2, and increased the levels of proapoptotic proteins, Bax and Bad. In addition, panaxydol inhibited the activation of Akt and extracellular signal-regulated kinase (ERK) and activated the p38 mitogen-activated protein kinase kinase (MAPK). Our results suggest that panaxydol is an anti-tumor compound that causes p53-mediated cell cycle arrest and apoptosis via mitochondrial apoptotic pathway in various cancer cells.

Keywords: apoptosis, cancer, G1 arrest, panaxydol

Procedia PDF Downloads 322