Search results for: differential gene expression

Authors: Marcel Verwey, Anna M. Joubert, Elsie M. Nolte, Wolfgang Dohle, Barry V. L. Potter, Anne E. Theron

Abstract:

A tetrahydroisoquinolinone (THIQ) core can be used to mimic the A,B-ring of colchicine site-binding microtubule disruptors such as 2-methoxyestradiol in the design of anti-cancer agents. Steroidomimeric microtubule disruptors were synthesized by introducing C'2 and C'3 of the steroidal A-ring to C'6 and C'7 of the THIQ core and by introducing a decorated hydrogen bond acceptor motif projecting from the steroidal D-ring to N'2. For this in vitro study, four non-steroidal THIQ-based analogues were investigated and comparative studies were done between the non-sulphamoylated compound STX 3450 and the sulphamoylated compounds STX 2895, STX 3329 and STX 3451. The objective of this study was to investigate the modes of cell death induced by these four THIQ-based analogues in A549 lung carcinoma epithelial cells and metastatic breast adenocarcinoma MDA-MB-231 cells. Cytotoxicity studies to determine the half maximal growth inhibitory concentrations were done using spectrophotometric quantification via crystal violet staining and lactate dehydrogenase (LDH) assays. Microtubule integrity and morphologic changes of exposed cells were investigated using polarization-optical transmitted light differential interference contrast microscopy, transmission electron microscopy and confocal microscopy. Flow cytometric quantification was used to determine apoptosis induction and the effect that THIQ-based analogues have on cell cycle progression. Signal transduction pathways were elucidated by quantification of the mitochondrial membrane integrity, cytochrome c release and caspase 3, -6 and -8 activation. Induction of autophagic cell death by the THIQ-based analogues was investigated by morphological assessment of fluorescent monodansylcadaverine (MDC) staining of acidic vacuoles and by quantifying aggresome formation via flow cytometry. Results revealed that these non-steroidal microtubule disrupting analogues inhibited 50% of cell growth at nanomolar concentrations. Immunofluorescence microscopy indicated microtubule depolarization and the resultant mitotic arrest was further confirmed through cell cycle analysis. Apoptosis induction via the intrinsic pathway was observed due to depolarization of the mitochondrial membrane, induction of cytochrome c release as well as, caspase 3 activation. Potential involvement of programmed cell death type II was observed due to the presence of acidic vacuoles and aggresome formation. Necrotic cell death did not contribute significantly, indicated by stable LDH levels. This in vitro study revealed the induction of the intrinsic apoptotic pathway as well as possible involvement of autophagy after exposure to these THIQ-based analogues in both MDA-MB-231- and A549 cells. Further investigation of this series of anticancer drugs still needs to be conducted to elucidate the temporal, mechanistic and functional crosstalk mechanisms between the two observed programmed cell deaths pathways.

Keywords: apoptosis, autophagy, cancer, microtubule disruptor

Procedia PDF Downloads 226

Authors: Tashwill Esterhuizen

Abstract:

Throughout Africa, several countries continue to have laws which criminalise same-sex sexual activities, which increases the vulnerability of the LGBT community to stigma, discrimination, and persecution. These criminal provisions often form the basis upon which states deny LGBT activists the right to freely associate with other like-minded individuals and form organizations that protect their interests and advocate for the rights and aspirations of the LGBT community. Over the past year, however, there has been significant progress in the advancement of universal, fundamental rights of LGBT persons throughout Africa. In many instances, these advancements came about through the bravery of activists who have publically insisted (in environments where same-sex sexual practices are criminalised) that their rights should be respected. Where meaningful engagement with the State was fruitless, activists took their plight to the judiciary and have successfully sought to uphold the fundamental rights of LGBT persons, paving the way for a more inclusive and tolerant society. Litigation Progress: Botswana is a prime example. For several years, the State denied a group of LGBT activists their right to freely associate and form their organisation Lesbians, Gays, and Bisexuals of Botswana (LEGABIBO), which aimed to promote the interests of the LGBT community in Botswana. In March 2016, the Botswana Court of Appeal found that the government’s refusal to register LEGABIBO violated the activists’ right to associate freely. The Court held that the right freedom of association applies to all persons regardless of their sexual orientation or gender identity. It does not matter that the views of the organisation are unpopular or unacceptable amongst the majority. In particular, the Court rejected the government of Botswana’s contention that registering LEGABIBO would disturb public peace and is contrary to public morality. Quite remarkably, the Court of Appeal recognised that while LGBT individuals are a minority group within the country, they are nonetheless persons entitled to constitutional protections of their dignity, regardless of whether they are unacceptable to others on religious or any other grounds. Furthermore, the Court held that human rights and fundamental freedoms are granted to all, including criminals or social outcasts because the denial of an individual’s humanity is the denial of their human dignity. This is crucial observation by the Court of Appeal, as once it is accepted that human rights apply to all human beings, then it becomes much easier for vulnerable groups to assert their own rights. Conclusion: The Botswana Court of Appeal decision, therefore, represents significant progress in the promotion of the rights of lesbian, gay, bisexual and transgender persons. The judgment has broader implications for many other countries which do not provide recognition of sexual minorities. It highlights the State’s duty to uphold basic rights and to ensure dignity, tolerance, and acceptance for marginalised persons.

Keywords: acceptance, freedom of association, freedom of expression, fundamental rights and freedoms, gender identity, human rights are universal, inclusive, inherent human dignity, progress, sexual orientation, tolerance

Procedia PDF Downloads 210

Authors: Kalyani Gandhi

Abstract:

The eighteenth century in Britain saw a great burst of activity in writing (letters, journals, newspapers, essays); often these modes of writing had a public-spirited bent in-step with the prevailing intellectual atmosphere. Mary Wollstonecraft was one of the leading intellectuals of that period who utilized letter writing to convey her thoughts on the exciting political developments of the late eighteenth century. Fusing together her anxieties and concerns about humanity in general and herself in particular, Wollstonecraft’s views of the world around her are filtered through the lens of her subjectivity. Thus, Wollstonecraft’s letters covered a wide range of topics on both the personal and political level (for the two are often entwined in Wollstonecraft’s characteristic style of analysis) such as sentiment, gender, nature, peasantry, the class system, the legal system, political duties and rights of both rulers and subjects, death, immortality, religion, family and education. Therefore, this paper intends to examine the manner in which Wollstonecraft utilizes letter-writing to constitute and develop Romantic self-hood, understand the world around her and illustrate her ideas on the political and social happenings in Europe. The primary text analyzed will be Mary Wollstonecraft's Letters Written During a Short Stay in Sweden, Denmark and Norway (1796) and the analysis of this text will be supplemented by researching 18th-century British letter writing culture, with a special emphasis on the epistolary habits of women. Within this larger framework, this paper intends to examine the manner in which this hybrid of travel and epistolary writing aided Mary Wollstonecraft's expression on Romantic selfhood and how it was complicated by ideas of gender. This paper reveals Wollstonecraft's text to be wrought with anxiety about the world around her and within her; thus, the personal-public nature of the epistolary format particularly suits her characteristic point of view that looks within and without. That is to say, Wollstonecraft’s anxieties about gender and self, are as much about the women she sees in the world around her as much as they are about her young daughter and herself. Wollstonecraft constantly explores and examines this anxiety within the different but interconnected realms of politics, economics, history and society. In fact, it is her complex technique of entwining these aforementioned concerns with a closer look at interpersonal relationships among men and women (she often mentions specific anecdotes and instances) that make Wollstonecraft's Letters so engaging and insightful. Thus, Wollstonecraft’s Letters is an exemplar of British Romantic writing due to the manner in which it explores the bond between the individual and society. Mary Wollstonecraft's nuances this exploration by incorporating her concerns about women and the playing out of gender in society. Thus, Wollstonecraft’s Letters is an invaluable contribution to the field of British Romanticism, particularly as it offers crucial insight on female Romantic writing that can broaden and enrich the current academic understanding of the field.

Keywords: British romanticism, letters, feminism, travel writing

Procedia PDF Downloads 185

Authors: S. Cichosz, A. Masek

Abstract:

A new cellulose hybrid modification approach, which is undoubtedly a scientific novelty, is introduced. The study reports the properties of cellulose (Arbocel UFC100 – Ultra Fine Cellulose) and characterizes cellulose filled polymer composites based on an ethylene-norbornene copolymer (TOPAS Elastomer E-140). Moreover, the approach of physicochemical two-stage cellulose treatment is introduced: solvent exchange (to ethanol or hexane) and further chemical modification with maleic anhydride (MA). Furthermore, the impact of the drying process on cellulose properties was investigated. Suitable measurements were carried out to characterize cellulose fibers: spectroscopic investigation (Fourier Transform Infrared Spektrofotometer-FTIR, Near InfraRed spectroscopy-NIR), thermal analysis (Differential scanning calorimetry, Thermal gravimetric analysis ) and Karl Fischer titration. It should be emphasized that for all UFC100 treatments carried out, a decrease in moisture content was evidenced. FT-IR reveals a drop in absorption band intensity at 3334 cm-1, the peak is associated with both –OH moieties and water. Similar results were obtained with Karl Fischer titration. Based on the results obtained, it may be claimed that the employment of ethanol contributes greatly to the lowering of cellulose water absorption ability (decrease of moisture content to approximately 1.65%). Additionally, regarding polymer composite properties, crucial data has been obtained from the mechanical and thermal analysis. The highest material performance was noted in the case of the composite sample that contained cellulose modified with MA after a solvent exchange with ethanol. This specimen exhibited sufficient tensile strength, which is almost the same as that of the neat polymer matrix – in the region of 40 MPa. Moreover, both the Payne effect and filler efficiency factor, calculated based on dynamic mechanical analysis (DMA), reveal the possibility of the filler having a reinforcing nature. What is also interesting is that, according to the Payne effect results, fibers dried before the further chemical modification are assumed to allow more regular filler structure development in the polymer matrix (Payne effect maximum at 1.60 MPa), compared with those not dried (Payne effect in the range 0.84-1.26 MPa). Furthermore, taking into consideration the data gathered from DSC and TGA, higher thermal stability is obtained in case of the materials filled with fibers that were dried before the carried out treatments (degradation activation energy in the region of 195 kJ/mol) in comparison with the polymer composite samples filled with unmodified cellulose (degradation activation energy of approximately 180 kJ/mol). To author’s best knowledge this work results in the introduction of a novel, new filler hybrid treatment approach. Moreover, valuable data regarding the properties of composites filled with cellulose fibers of various moisture contents have been provided. It should be emphasized that plant fiber-based polymer bio-materials described in this research might contribute significantly to polymer waste minimization because they are more readily degraded.

Keywords: cellulose fibers, solvent exchange, moisture content, ethylene-norbornene copolymer

Procedia PDF Downloads 97

Authors: Çağrı Bekir Baysal, Göktuğ Burak Çalık

Abstract:

Armored vehicles may have different traction layouts as a result of terrain capabilities and mobility needs. Two main categories of layouts can be separated as wheeled and tracked. Tracked vehicles have superior off-road capabilities but what they gain on terrain performance they lose on mobility front. Wheeled vehicles on the other hand do not have as good terrain capabilities as tracked vehicles but they have superior mobility capabilities such as top speed, range and agility with respect to tracked vehicles. Conventional armored vehicles employ a diesel ICE as main power source. In these vehicles ICE is mechanically connected to the powertrain. This determines the ICE rpm as a result of speed and torque requested by the driver. ICE efficiency changes drastically with torque and speed required and conventional vehicles suffer in terms of fuel consumption because of this. Hybrid electric vehicles employ at least one electric motor in order to improve fuel efficiency. There are different types of hybrid vehicles but main types are Series Hybrid, Parallel Hybrid and Series-Parallel Hybrid. These vehicles introduce an electric motor for traction and also can have a generator electric motor for range extending purposes. Having an electric motor as the traction power source brings the flexibility of either using the ICE as an alternative traction source while it is in efficient range or completely separating the ICE from traction and using it solely considering efficiency. Hybrid configurations have additional advantages for armored vehicles in addition to fuel efficiency. Heat signature, silent operation and prolonged stationary missions can be possible with the help of the high-power battery pack that will be present in the vehicle for hybrid drivetrain. Because of the reasons explained, hybrid armored vehicles are becoming a target area for military and also for vehicle suppliers. In order to have a better idea and starting point when starting a hybrid armored vehicle design, hybrid drivetrain configuration has to be selected after performing a trade-off study. This study has to include vehicle mobility simulations, integration level, vehicle level and performance level criteria. In this study different hybrid traction configurations possible for an 8x8 vehicle is compared using above mentioned criteria set. In order to compare hybrid traction configurations ease of application, cost, weight advantage, reliability, maintainability, redundancy and performance criteria have been used. Performance criteria points have been defined with the help of vehicle simulations and tests. Results of these simulations and tests also help determining required tractive power for an armored vehicle including conditions like trench and obstacle crossing, gradient climb. With the method explained in this study, each configuration is assigned a point for each criterion. This way, correct configuration can be selected objectively for every application. Also, key aspects of armored vehicles, mine protection and ballistic protection will be considered for hybrid configurations. Results are expected to vary for different types of vehicles but it is observed that having longitudinal differential locking capability improves mobility and having high motor count increases complexity in general.

Keywords: armored vehicles, electric drivetrain, electric mobility, hybrid vehicles

Procedia PDF Downloads 62

Authors: Yazan S. Khaled, Shazana Shamsuddin, Jim Tiernan, Mike McPherson, Thomas Hughes, Paul Millner, David G. Jayne

Abstract:

Introduction: There is a need for real-time imaging of colorectal cancer (CRC) to allow tailored surgery to the disease stage. Fluorescence guided laparoscopic imaging of primary colorectal cancer and the draining lymphatics would potentially bring stratified surgery into clinical practice and realign future CRC management to the needs of patients. Fluorescent nanoparticles can offer many advantages in terms of intra-operative imaging and therapy (theranostic) in comparison with traditional soluble reagents. Nanoparticles can be functionalised with diverse reagents and then targeted to the correct tissue using an antibody or Affimer (artificial binding protein). We aimed to develop and test fluorescent silica nanoparticles and targeted against CRC using an anti-carcinoembryonic antigen (CEA) Affimer (Aff). Methods: Anti-CEA and control Myoglobin Affimer binders were subcloned into the expressing vector pET11 followed by transformation into BL21 Star™ (DE3) E.coli. The expression of Affimer binders was induced using 0.1 mM isopropyl β-D-1-thiogalactopyranoside (IPTG). Cells were harvested, lysed and purified using nickle chelating affinity chromatography. The photosensitiser Foslip (soluble analogue of 5,10,15,20-Tetra(m-hydroxyphenyl) chlorin) was incorporated into the core of silica nanoparticles using water-in-oil microemulsion technique. Anti-CEA or control Affs were conjugated to silica nanoparticles surface using sulfosuccinimidyl-4-(N-maleimidomethyl) cyclohexane-1-carboxylate (sulfo SMCC) chemical linker. Binding of CEA-Aff or control nanoparticles to colorectal cancer cells (LoVo, LS174T and HC116) was quantified in vitro using confocal microscopy. Results: The molecular weights of the obtained band of Affimers were ~12.5KDa while the diameter of functionalised silica nanoparticles was ~80nm. CEA-Affimer targeted nanoparticles demonstrated 9.4, 5.8 and 2.5 fold greater fluorescence than control in, LoVo, LS174T and HCT116 cells respectively (p < 0.002) for the single slice analysis. A similar pattern of successful CEA-targeted fluorescence was observed in the maximum image projection analysis, with CEA-targeted nanoparticles demonstrating 4.1, 2.9 and 2.4 fold greater fluorescence than control particles in LoVo, LS174T, and HCT116 cells respectively (p < 0.0002). There was no significant difference in fluorescence for CEA-Affimer vs. CEA-Antibody targeted nanoparticles. Conclusion: We are the first to demonstrate that Foslip-doped silica nanoparticles conjugated to anti-CEA Affimers via SMCC allowed tumour cell-specific fluorescent targeting in vitro, and had shown sufficient promise to justify testing in an animal model of colorectal cancer. CEA-Affimer appears to be a suitable targeting molecule to replace CEA-Antibody. Targeted silica nanoparticles loaded with Foslip photosensitiser is now being optimised to drive photodynamic killing, via reactive oxygen generation.

Keywords: colorectal cancer, silica nanoparticles, Affimers, antibodies, imaging

Procedia PDF Downloads 216

Authors: Johns Paul, Santhosh J. Nalluveettil, P. Purushothaman, M. Premdas

Abstract:

Lunar landing is one of the most critical phases of lunar mission. The lander is provided with a soft landing system to prevent structural damage of lunar module by absorbing the landing shock and also assure stability during landing. Presently available software are not capable to simulate the rigid body dynamics coupled with contact simulation and elastic/plastic deformation analysis. Hence a separate mathematical model has been generated for studying the dynamics of a typical lunar soft lander. Parameters used in the analysis includes lunar surface slope, coefficient of friction, initial touchdown velocity (vertical and horizontal), mass and moment of inertia of lander, crushing force due to energy absorbing material in the legs, number of legs and geometry of lander. The mathematical model is capable to simulate plastic and elastic deformation of honey comb, frictional force between landing leg and lunar soil, surface contact simulation, lunar gravitational force, rigid body dynamics and linkage dynamics of inverted tripod landing gear. The non linear differential equations generated for studying the dynamics of lunar lander is solved by numerical method. Matlab programme has been used as a computer tool for solving the numerical equations. The position of each kinematic joint is defined by mathematical equations for the generation of equation of motion. All hinged locations are defined by position vectors with respect to body fixed coordinate. The vehicle rigid body rotations and motions about body coordinate are only due to the external forces and moments arise from footpad reaction force due to impact, footpad frictional force and weight of vehicle. All these force are mathematically simulated for the generation of equation of motion. The validation of mathematical model is done by two different phases. First phase is the validation of plastic deformation of crushable elements by employing conservation of energy principle. The second phase is the validation of rigid body dynamics of model by simulating a lander model in ADAMS software after replacing the crushable elements to elastic spring element. Simulation of plastic deformation along with rigid body dynamics and contact force cannot be modeled in ADAMS. Hence plastic element of primary strut is replaced with a spring element and analysis is carried out in ADAMS software. The same analysis is also carried out using the mathematical model where the simulation of honeycomb crushing is replaced by elastic spring deformation and compared the results with ADAMS analysis. The rotational motion of linkages and 6 degree of freedom motion of lunar Lander about its CG can be validated by ADAMS software by replacing crushing element to spring element. The model is also validated by the drop test results of 4 leg lunar lander. This paper presents the details of mathematical model generated and its validation.

Keywords: honeycomb, landing leg tripod, lunar lander, primary link, secondary link

Procedia PDF Downloads 323

Authors: Shamini Dhana, G. Subrahmanya VRK Rao

Abstract:

The fashion industry is advancing towards a mindset of zero waste, personalization, creativity, and circularity. The trend of upcycling clothing and materials into personalized fashion is being demanded by the next generation. There is a need for a digital tool to accelerate the process towards mass customization. Dhana’s D/Sphere fashion technology platform uses digital tools to accelerate upcycling. In essence, advanced fashion garments can be designed and developed via reuse, repurposing, recreating activities, and using existing fabric and circulating materials. The D/Sphere platform has the following objectives: to provide (1) An opportunity to develop modern fashion using existing, finished materials and clothing without chemicals or water consumption; (2) The potential for an everyday customer and designer to use the medium of fashion for creative expression; (3) A solution to address the global textile waste generated by pre- and post-consumer fashion; (4) A solution to reduce carbon emissions, water, and energy consumption with the participation of all stakeholders; (5) An opportunity for brands, manufacturers, retailers to work towards zero-waste designs and as an alternative revenue stream. Other benefits of this alternative approach include sustainability metrics, trend prediction, facilitation of disassembly and remanufacture deep learning, and hyperheuristics for high accuracy. A design tool for mass personalization and customization utilizing existing circulating materials and deadstock, targeted to fashion stakeholders will lower environmental costs, increase revenues through up to date upcycled apparel, produce less textile waste during the cut-sew-stitch process, and provide a real design solution for the end customer to be part of circular fashion. The broader impact of this technology will result in a different mindset to circular fashion, increase the value of the product through multiple life cycles, find alternatives towards zero waste, and reduce the textile waste that ends up in landfills. This technology platform will be of interest to brands and companies that have the responsibility to reduce their environmental impact and contribution to climate change as it pertains to the fashion and apparel industry. Today, over 70% of the $3 trillion fashion and apparel industry ends up in landfills. To this extent, the industry needs such alternative techniques to both address global textile waste as well as provide an opportunity to include all stakeholders and drive circular fashion with new personalized products. This type of modern systems thinking is currently being explored around the world by the private sector, organizations, research institutions, and governments. This technological innovation using digital tools has the potential to revolutionize the way we look at communication, capabilities, and collaborative opportunities amongst stakeholders in the development of new personalized and customized products, as well as its positive impacts on society, our environment, and global climate change.

Keywords: circular fashion, deep learning, digital technology platform, personalization

Procedia PDF Downloads 34

Authors: Maria de Fátima Lambert

Abstract:

Bernardo Soares is one of Fernando Pessoa' several heteronyms (and "half-heteronyms"). Perhaps the one that brought together the majority of his qualities and characters of self-identity within the famous inner-persona-alter-diversity. The Book of Disquiet by Bernardo Soares was released in 1983, consisting of ontological remarks caught by an obsessive inquiring about self-existence. The book became a highly valuable substance when focusing upon the philosophical grounds of Pessoa's aesthetics. For sure, we cannot consider a single aesthetic, admitting that each heteronym has its own particular one, developed after different principles and convictions. Regarding Bernardo Soares, his thought arises from sequenced self-clues expressing peculiar existential doubtless presented as certainties -and vice-versa. His written self-search-images are reported, molding the painful awareness of existence through the discredited tolerance of any conclusive dialogue with others. Given the nature of Soares’ [maybe] unfinished writings, it is obvious that he headed far from his self-insurance-capsule: the office, bedroom, or even the walkscapes through Lisbon. The idea of travel/journey is one of the most relevant when recognizing his profound - although undercover - anguish as melancholy and nostalgia. In Bernardo Soares, Aesthetics is taken agonizingly, grounded upon discreet poetic phraseology and terms. His poetical awareness developed compulsive titles such "Aesthetics of Indifference", "Aesthetics of Discouragement". Soares' Aesthetics emerges directly from oneself, understanding art as inner acts and living experienced issues. Art is not freed from the intellectual expression of oneself emotions. The Disquiet Book is an existential nightmare nourished by everyday life, single written thoughts, balanced by melancholia, nostalgia, and distress. One might wonder if it was dreams that guided his fictional literary persona or the narrow facts of life itself. Along with his endless disquiet writing, Pessoa’s semi-heteronymous traveled without physically going anywhere. The complexity of inner existence is fulfilled by lonely mental walks and travels, as in two texts titled The Never Accomplished Journey. Although we also can consider other fragments, these are the deepest reflections about travelling. Let’s recall that Fernando Pessoa’s ortonyms writings -poems and essays- also addressed this issue from a philosophical perspective. We believe that this theme is one of the meaningful concepts for featuring the main principles of his aesthetics. As we know, Fernando Pessoa did not travel to foreign countries (or in Portugal), except for the journey, with his family, from Lisbon to South Africa (as a child) and, some years later, the return back to Lisbon. One may wonder why the poet never undertook other journeys. Maybe due to a disbelief in moving away from his comfort zone or due to the fear of becoming addicted to endless travels and the loss of his convenient self-closeness. In The Book of Disquiet, the poet shared his internal visions of the outer world but mainly visualizing his deepest enigmas and experiences -so strongly incorporated into reality and fiction.

Keywords: aesthetic principles, Bernardo Soares, Fernando Pessoa , melancholia, nostalgia, non-accomplished travel, The Book of Disquiet

Procedia PDF Downloads 99

Authors: Elena Filova, Ondrej Kaplan, Marie Markova, Helena Dragounova, Roman Matejka, Eduard Brynda, Lucie Bacakova

Abstract:

Decellularized vessels have been evaluated as small-diameter vascular prostheses. Reseeding autologous cells onto decellularized tissue prior implantation should prolong prostheses function and make them living tissues. Suitable cell types for reseeding are both endothelial cells and bone marrow-derived stem cells, with a capacity for differentiation into smooth muscle cells upon mechanical loading. Endothelial cells assure antithrombogenicity of the vessels and MSCs produce growth factors and, after their differentiation into smooth muscle cells, they are contractile and produce extracellular matrix proteins as well. Fibrin is a natural scaffold, which allows direct cell adhesion based on integrin receptors. It can be prepared autologous. Fibrin can be modified with bound growth factors, such as basic fibroblast growth factor (FGF-2) and vascular endothelial growth factor (VEGF). These modifications in turn make the scaffold more attractive for cells ingrowth into the biological scaffold. The aim of the study was to prepare thin surface-attached fibrin assemblies with bound FGF-2 and VEGF, and to evaluate growth and differentiation of bone marrow-derived mesenchymal stem cells on the fibrin (Fb) assemblies. Following thin surface-attached fibrin assemblies were prepared: Fb, Fb+VEGF, Fb+FGF2, Fb+heparin, Fb+heparin+VEGF, Fb+heparin+FGF2, Fb+heparin+FGF2+VEGF. Cell culture poly-styrene and glass coverslips were used as controls. Human MSCs (passage 3) were seeded at the density of 8800 cells/1.5 mL alpha-MEM medium with 2.5% FS and 200 U/mL aprotinin per well of a 24-well cell culture. The cells have been cultured on the samples for 6 days. Cell densities on day 1, 3, and 6 were analyzed after staining with LIVE/DEAD cytotoxicity/viability assay kit. The differentiation of MSCs is being analyzed using qPCR. On day 1, the highest density of MSCs was observed on Fb+VEGF and Fb+FGF2. On days 3 and 6, there were similar densities on all samples. On day 1, cell morphology was polygonal and spread on all sample. On day 3 and 6, MSCs growing on Fb assemblies with FGF2 became apparently elongated. The evaluation of expression of genes for von Willebrand factor and CD31 (endothelial cells), for alpha-actin (smooth muscle cells), and for alkaline phosphatase (osteoblasts) is in progress. We prepared fibrin assemblies with bound VEGF and FGF-2 that supported attachment and growth of mesenchymal stem cells. The layers are promising for improving the ingrowth of MSCs into the biological scaffold. Supported by the Technology Agency of the Czech Republic TA04011345, and Ministry of Health NT11270-4/2010, and BIOCEV – Biotechnology and Biomedicine Centre of the Academy of Sciences and Charles University” project (CZ.1.05/1.1.00/02.0109), funded by the European Regional Development Fund for their financial supports.

Keywords: fibrin assemblies, FGF-2, mesenchymal stem cells, VEGF

Procedia PDF Downloads 301

Authors: Alvyra Galkienė, Ona Monkevičienė

Abstract:

The full participation of all pupils in the overall educational process is defined by the concept of inclusive education, which is gradually evolving in education policy and practice. It includes the full participation of all pupils in a shared learning experience and educational practices that address barriers to learning. Inclusive education applying the principles of Universal Design for Learning (UDL), which includes promoting students' involvement in learning processes, guaranteeing a deep understanding of the analysed phenomena, initiating self-directed learning, and using e-tools to create a barrier-free environment, is a prerequisite for the personal success of each pupil. However, the sustainability of quality education is affected by the transformation of education systems. This was particularly evident during the period of the forced transition from contact to distance education in the COVID-19 pandemic. Research Problem: The transformation of the educational environment from real to virtual one and the loss of traditional forms of educational support highlighted the need for new research, revealing the individual profiles of teachers using UDL-based learning and the pathways of sustainable transfer of successful practices to non-conventional learning environments. Research Methods: In order to identify individual latent teacher profiles that encompass the essential components of UDL-based inclusive teaching and direct leadership of students' learning, the quantitative analysis software Mplius was used for latent profile analysis (LPA). In order to reveal proven, i.e., sustainable, pathways for the transit of the components of UDL-based inclusive learning to distance learning, latent profile transit analysis (LPTA) via Mplius was used. An online self-reported questionnaire was used for data collection. It consisted of blocks of questions designed to reveal the experiences of subject teachers in contact and distance learning settings. 1432 Lithuanian, Latvian, and Estonian subject teachers took part in the survey. Research Results: The LPA analysis revealed eight latent teacher profiles with different characteristics of UDL-based inclusive education or traditional teaching in contact teaching conditions. Only 4.1% of the subject teachers had a profile characterised by a sustained UDL approach to teaching: promoting pupils' self-directed learning; empowering pupils' engagement, understanding, independent action, and expression; promoting pupils' e-inclusion; and reducing the teacher's direct supervision of the students. Other teacher profiles were characterised by limited UDL-based inclusive education either due to the lack of one or more of its components or to the predominance of direct teacher guidance. The LPTA analysis allowed us to highlight the following transit paths of teacher profiles in the extreme conditions of the transition from contact to distance education: teachers staying in the same profile of UDL-based inclusive education (sustainable transit) or jumping to other profiles (unsustainable transit in case of barriers), and teachers from other profiles moving to this profile (ongoing transit taking advantage of the changed new possibilities in the teaching process).

Keywords: distance education, latent teacher profiles, sustainable transit, UDL

Procedia PDF Downloads 71

Authors: Archana Sharma, Vijayashree Nayak, Ashok Kumar

Abstract:

Three-dimensional matrices were fabricated from blend of natural-natural polymers like carrageenan-gelatin and synthetic -natural polymers such as PEG- gelatin (PEG of different molecular weights (2,000 and 6,000) using two different crosslinkers; glutaraldehyde and EDC-NHS by cryogelation technique. Blends represented a feasible approach to design 3-D scaffolds with controllable mechanical, physical and biochemical properties without compromising biocompatibility and biodegradability. These matrices possessed interconnected porous structure, good mechanical strength, biodegradable nature, constant swelling kinetics, ability to withstand high temperature and visco-elastic behavior. Hemocompatibility of cryogel matrices was determined by coagulation assays and hemolytic activity assay which demonstrated that these cryogels have negligible effects on coagulation time and have excellent blood compatibility. In vitro biocompatibility (cell-matrix interaction) inferred good cell adhesion, proliferation, and secretion of ECM on matrices. These matrices provide a microenvironment for the growth, proliferation, differentiation and secretion of ECM of different cell types such as IMR-32, C2C12, Cos-7, rat bone marrow derived MSCs and human bone marrow MSCs. Hoechst 33342 and PI staining also confirmed that the cells were uniformly distributed, adhered and proliferated properly on the cryogel matrix. An ideal scaffold used for tissue engineering application should allow the cells to adhere, proliferate and maintain their functionality. Neurotransmitter analysis has been done which indicated that IMR-32 cells adhered, proliferated and secreted neurotransmitters when they interacted with these matrices which showed restoration of their functionality. The cell-matrix interaction up to molecular level was also evaluated so to check genotoxicity and protein expression profile which indicated that these cryogel matrices are non-genotoxic and maintained biofunctionality of cells growing on these matrices. All these cryogels, when implanted subcutaneously in balb/c mice, showed no adverse systemic or local toxicity effects at implantation site. There was no significant increase in inflammatory cell count has otherwise been observed after scaffold implantation. These cryogels are supermacroporous and this porous structure allows cell infiltration and proliferation of host cells. This showed the integration and presence of infiltrated cells into the cryogel implants. Histological analysis confirmed that the implanted cryogels do not have any adverse effect in spite of host immune system recognition at the site of implantation, on its surrounding tissues and other vital host organs. In vivo biocompatibility study after in vitro biocompatibility analysis has also concluded that these synthesized cryogels act as important biological substitutes, more adaptable and appropriate for transplantation. Thus, these cryogels showed their potential for soft tissue engineering applications.

Keywords: cryogelation, hemocompatibility, in vitro biocompatibility, in vivo biocompatibility, soft tissue engineering applications

Procedia PDF Downloads 197

Authors: Anwar Beg, Sireetorn Kuharat, Rashid Mehmood, Rabil Tabassum, Meisam Babaie

Abstract:

Nano-polymeric solar paints and sol-gels have emerged as a major new development in solar cell/collector coatings offering significant improvements in durability, anti-corrosion and thermal efficiency. They also exhibit substantial viscosity variation with temperature which can be exploited in solar collector designs. Modern manufacturing processes for such nano-rheological materials frequently employ stagnation flow dynamics under high temperature which invokes radiative heat transfer. Motivated by elaborating in further detail the nanoscale heat, mass and momentum characteristics of such sol gels, the present article presents a mathematical and computational study of the steady, two-dimensional, non-aligned thermo-fluid boundary layer transport of copper metal-doped water-based nano-polymeric sol gels under radiative heat flux. To simulate real nano-polymer boundary interface dynamics, thermal slip is analysed at the wall. A temperature-dependent viscosity is also considered. The Tiwari-Das nanofluid model is deployed which features a volume fraction for the nanoparticle concentration. This approach also features a Maxwell-Garnet model for the nanofluid thermal conductivity. The conservation equations for mass, normal and tangential momentum and energy (heat) are normalized via appropriate transformations to generate a multi-degree, ordinary differential, non-linear, coupled boundary value problem. Numerical solutions are obtained via the stable, efficient Runge-Kutta-Fehlberg scheme with shooting quadrature in MATLAB symbolic software. Validation of solutions is achieved with a Variational Iterative Method (VIM) utilizing Langrangian multipliers. The impact of key emerging dimensionless parameters i.e. obliqueness parameter, radiation-conduction Rosseland number (Rd), thermal slip parameter (α), viscosity parameter (m), nanoparticles volume fraction (ϕ) on non-dimensional normal and tangential velocity components, temperature, wall shear stress, local heat flux and streamline distributions is visualized graphically. Shear stress and temperature are boosted with increasing radiative effect whereas local heat flux is reduced. Increasing wall thermal slip parameter depletes temperatures. With greater volume fraction of copper nanoparticles temperature and thermal boundary layer thickness is elevated. Streamlines are found to be skewed markedly towards the left with positive obliqueness parameter.

Keywords: non-orthogonal stagnation-point heat transfer, solar nano-polymer coating, MATLAB numerical quadrature, Variational Iterative Method (VIM)

Procedia PDF Downloads 109

Authors: Elyria Kemp, Kelly Cowart, My Bui

Abstract:

According to the National Institute of Mental Health, an estimated 31.9% of adolescents have had an anxiety disorder. Several environmental factors may help to contribute to high levels of anxiety and depression in young people (i.e., Generation Z, Millennials). However, as young people negotiate life on social media, they may begin to evaluate themselves using excessively high standards and adopt self-perfectionism tendencies. Broadly defined, self-perfectionism involves very critical evaluations of the self. Perfectionism may also come from others and may manifest as socially prescribed perfectionism, and young adults are reporting higher levels of socially prescribed perfectionism than previous generations. This rising perfectionism is also associated with anxiety, greater physiological reactivity, and a sense of social disconnection. However, theories from psychology suggest that improvement in emotion regulation can contribute to enhanced psychological and emotional well-being. Emotion regulation refers to the ways people manage how and when they experience and express their emotions. Cognitive reappraisal and expressive suppression are common emotion regulation strategies. Cognitive reappraisal involves changing the meaning of a stimulus that involves construing a potentially emotion-eliciting situation in a way that changes its emotional impact. By contrast, expressive suppression involves inhibiting the behavioral expression of emotion. The purpose of this research is to examine the efficacy of social marketing initiatives which promote emotion regulation strategies to help young adults regulate their emotions. In Study 1 a single factor (emotional regulation strategy: a cognitive reappraisal, expressive, control) between-subjects design was conducted using an online, non-student consumer panel (n=96). Sixty-eight percent of participants were male, and 32% were female. Study participants belonged to the Millennial and Gen Z cohort, ranging in age from 22 to 35 (M=27). Participants were first told to spend at least three minutes writing about a public speaking appearance which made them anxious. The purpose of this exercise was to induce anxiety. Next, participants viewed one of three advertisements (randomly assigned) which promoted an emotion regulation strategy—cognitive reappraisal, expressive suppression, or an advertisement non-emotional in nature. After being exposed to one of the ads, participants responded to a measure composed of two items to access their emotional state and the efficacy of the messages in fostering emotion management. Findings indicated that individuals in the cognitive reappraisal condition (M=3.91) exhibited the most positive feelings and more effective emotion regulation than the expressive suppression (M=3.39) and control conditions (M=3.72, F(1,92) = 3.3, p<.05). Results from this research can be used by institutions (e.g., schools) in taking a leadership role in attacking anxiety and other mental health issues. Social stigmas regarding mental health can be removed and a more proactive stance can be taken in promoting healthy coping behaviors and strategies to manage negative emotions.

Keywords: emotion regulation, anxiety, social marketing, generation z

Procedia PDF Downloads 180

Authors: Zatovska T. V., Nesterova N. V., Baranova G. V., Zagorodnya S. D.

Abstract:

In recent years, biosensor technologies based on the phenomenon of surface plasmon resonance (SPR) are becoming increasingly used in biology and medicine. Their application facilitates exploration in real time progress of binding of biomolecules and identification of agents that specifically interact with biologically active substances immobilized on the biosensor surface (biochips). Special attention is paid to the use of Biosensor analysis in determining the antibody-antigen interaction in the diagnostics of diseases caused by viruses and bacteria. According to WHO, the diseases that are caused by the herpes simplex virus (HSV), take second place (15.8%) after influenza as a cause of death from viral infections. Current diagnostics of HSV infection include PCR and ELISA assays. The latter allows determination the degree of immune response to viral infection and respective stages of its progress. In this regard, the searches for new and available diagnostic methods are very important. This work was aimed to develop Biosensor chip for detection of specific antibodies to HSV-1 in the human blood serum. The proteins of HSV1 (strain US) were used as antigens. The viral particles were accumulated in cell culture MDBK and purified by differential centrifugation in cesium chloride density gradient. Analysis of the HSV1 proteins was performed by polyacrylamide gel electrophoresis and ELISA. The protein concentration was measured using De Novix DS-11 spectrophotometer. The device for detection of antigen-antibody interactions was an optoelectronic two-channel spectrometer ‘Plasmon-6’, using the SPR phenomenon in the Krechman optical configuration. It was developed at the Lashkarev Institute of Semiconductor Physics of NASU. The used carrier was a glass plate covered with 45 nm gold film. Screening of human blood serums was performed using the test system ‘HSV-1 IgG ELISA’ (GenWay, USA). Development of Biosensor chip included optimization of conditions of viral antigen sorption and analysis steps. For immobilization of viral proteins 0.2% solution of Dextran 17, 200 (Sigma, USA) was used. Sorption of antigen took place at 4-8°C within 18-24 hours. After washing of chip, three times with citrate buffer (pH 5,0) 1% solution of BSA was applied to block the sites not occupied by viral antigen. It was found direct dependence between the amount of immobilized HSV1 antigen and SPR response. Using obtained biochips, panels of 25 positive and 10 negative for the content of antibodies to HSV-1 human sera were analyzed. The average value of SPR response was 185 a.s. for negative sera and from 312 to. 1264 a.s. for positive sera. It was shown that SPR data were agreed with ELISA results in 96% of samples proving the great potential of SPR in such researches. It was investigated the possibility of biochip regeneration and it was shown that application of 10 mM NaOH solution leads to rupture of intermolecular bonds. This allows reuse the chip several times. Thus, in this study biosensor chip for detection of specific antibodies to HSV1 was successfully developed expanding a range of diagnostic methods for this pathogen.

Keywords: biochip, herpes virus, SPR

Procedia PDF Downloads 398

Authors: Mengjiao Fan, Thomson W. L. Wong

Abstract:

It is unavoidable that older adults may have to deal with aging-related motor problems. Aging is highly likely to affect motor learning and control as well. For example, older adults may suffer from poor motor function and quality of life due to age-related eye changes. These adverse changes in vision results in impairment of movement automaticity. Reaching is a fundamental component of various complex movements, which is therefore beneficial to explore the changes and adaptation in visuomotor behaviors. The current study aims to explore how aging affects visuomotor behaviors by comparing motor performance and gaze behaviors between two age groups (i.e., young and older adults). Visuomotor behaviors in reaching under providing or blocking online visual feedback (simulated visual deficiency) conditions were investigated in 60 healthy young adults (Mean age=24.49 years, SD=2.12) and 37 older adults (Mean age=70.07 years, SD=2.37) with normal or corrected-to-normal vision. Participants in each group were randomly allocated into two subgroups. Subgroup 1 was provided with online visual feedback of the hand-controlled mouse cursor. However, in subgroup 2, visual feedback was blocked to simulate visual deficiency. The experimental task required participants to complete 20 times of reaching to a target by controlling the mouse cursor on the computer screen. Among all the 20 trials, start position was upright in the center of the screen and target appeared at a randomly selected position by the tailor-made computer program. Primary outcomes of motor performance and gaze behaviours data were recorded by the EyeLink II (SR Research, Canada). The results suggested that aging seems to affect the performance of reaching tasks significantly in both visual feedback conditions. In both age groups, blocking online visual feedback of the cursor in reaching resulted in longer hand movement time (p < .001), longer reaching distance away from the target center (p<.001) and poorer reaching motor accuracy (p < .001). Concerning gaze behaviors, blocking online visual feedback increased the first fixation duration time in young adults (p<.001) but decreased it in older adults (p < .001). Besides, under the condition of providing online visual feedback of the cursor, older adults conducted a longer fixation dwell time on target throughout reaching than the young adults (p < .001) although the effect was not significant under blocking online visual feedback condition (p=.215). Therefore, the results suggested that different levels of visual feedback during movement execution can affect gaze behaviors differently in older and young adults. Differential effects by aging on visuomotor behaviors appear on two visual feedback patterns (i.e., blocking or providing online visual feedback of hand-controlled cursor in reaching). Several specific gaze behaviors among the older adults were found, which imply that blocking of visual feedback may act as a stimulus to seduce extra perceptive load in movement execution and age-related visual degeneration might further deteriorate the situation. It indeed provides us with insight for the future development of potential rehabilitative training method (e.g., well-designed errorless training) in enhancing visuomotor adaptation for our aging population in the context of improving their movement automaticity by facilitating their compensation of visual degeneration.

Keywords: aging effect, movement automaticity, reaching, visuomotor behaviors, visual degeneration

Procedia PDF Downloads 291

Authors: Timur S. Zaripov, Artur K. Gilfanov, Sergei S. Sazhin, Steven M. Begg, Morgan R. Heikal

Abstract:

The calculation of the concentration of inertial droplets in complex flows is encountered in the modelling of numerous engineering and environmental phenomena; for example, fuel droplets in internal combustion engines and airborne pollutant particles. The results of recent research, focused on the development of methods for calculating concentration and their implementation in the commercial CFD code, ANSYS Fluent, is presented here. The study is motivated by the investigation of the mixture preparation processes in internal combustion engines with direct injection of fuel sprays. Two methods are used in our analysis; the Fully Lagrangian method (also known as the Osiptsov method) and the Eulerian approach. The Osiptsov method predicts droplet concentrations along path lines by solving the equations for the components of the Jacobian of the Eulerian-Lagrangian transformation. This method significantly decreases the computational requirements as it does not require counting of large numbers of tracked droplets as in the case of the conventional Lagrangian approach. In the Eulerian approach the average droplet velocity is expressed as a function of the carrier phase velocity as an expansion over the droplet response time and transport equation can be solved in the Eulerian form. The advantage of the method is that droplet velocity can be found without solving additional partial differential equations for the droplet velocity field. The predictions from the two approaches were compared in the analysis of the problem of a dilute gas-droplet flow around an infinitely long, circular cylinder. The concentrations of inertial droplets, with Stokes numbers of 0.05, 0.1, 0.2, in steady-state and transient laminar flow conditions, were determined at various Reynolds numbers. In the steady-state case, flows with Reynolds numbers of 1, 10, and 100 were investigated. It has been shown that the results predicted using both methods are almost identical at small Reynolds and Stokes numbers. For larger values of these numbers (Stokes — 0.1, 0.2; Reynolds — 10, 100) the Eulerian approach predicted a wider spread in concentration in the perturbations caused by the cylinder that can be attributed to the averaged droplet velocity field. The transient droplet flow case was investigated for a Reynolds number of 200. Both methods predicted a high droplet concentration in the zones of high strain rate and low concentrations in zones of high vorticity. The maxima of droplet concentration predicted by the Osiptsov method was up to two orders of magnitude greater than that predicted by the Eulerian method; a significant variation for an approach widely used in engineering applications. Based on the results of these comparisons, the Osiptsov method has resulted in a more precise description of the local properties of the inertial droplet flow. The method has been applied to the analysis of the results of experimental observations of a liquid gasoline spray at representative fuel injection pressure conditions. The preliminary results show good qualitative agreement between the predictions of the model and experimental data.

Keywords: internal combustion engines, Eulerian approach, fully Lagrangian approach, gasoline fuel sprays, droplets and particle concentrations

Procedia PDF Downloads 234

Authors: Georgios Samaras

Abstract:

New media such as Facebook, YouTube and Twitter introduced the world to a new era of instant communication. An era where online interactions could replace a lot of offline actions. Technology can create a mediated environment in which participants can communicate (one-to-one, one-to-many, and many-to-many) both synchronously and asynchronously and participate in reciprocal message exchanges. Currently, social networks are attracting similar academic attention to that of the internet after its mainstream implementation into public life. Websites and platforms are seen as the forefront of a new political change. There is a significant backdrop of previous methodologies employed to research the effects of social networks. New approaches are being developed to be able to adapt to the growth of social networks and the invention of new platforms. Golden Dawn was the first openly neo-Nazi party post World War II to win seats in the parliament of a European country. Its racist rhetoric and violent tactics on social networks were rewarded by their supporters, who in the face of Golden Dawn’s leaders saw a ‘new dawn’ in Greek politics. Mainstream media banned its leaders and members of the party indefinitely after Ilias Kasidiaris attacked Liana Kanelli, a member of the Greek Communist Party, on live television. This media ban was seen as a treasonous move by a significant percentage of voters, who believed that the system was desperately trying to censor Golden Dawn to favor mainstream parties. The shocking attack on live television received international coverage and while European countries were condemning this newly emerged neo-Nazi rhetoric, almost 7 percent of the Greek population rewarded Golden Dawn with 18 seats in the Greek parliament. Many seem to think that Golden Dawn mobilised its voters online and this approach played a significant role in spreading their message and appealing to wider audiences. No strict online censorship existed back in 2012 and although Golden Dawn was openly used neo-Nazi symbolism, it was allowed to use social networks without serious restrictions until 2017. This paper used qualitative methods to investigate Golden Dawn’s rise in social networks from 2012 to 2019. The focus of the content analysis was set on three social networking platforms: Facebook, Twitter and YouTube, while the existence of Golden Dawn’s website, which was used as a news sharing hub, was also taken into account. The content analysis included text and visual analyses that sampled content from their social networking pages to translate their political messaging through an ideological lens focused on extreme-right populism. The absence of hate speech regulations on social network platforms in 2012 allowed the free expression of those heavily ultranationalist and populist views, as they were employed by Golden Dawn in the Greek political scene. On YouTube, Facebook and Twitter, the influence of their rhetoric was particularly strong. Official channels and MPs profiles were investigated to explore the messaging in-depth and understand its ideological elements.

Keywords: populism, far-right, social media, Greece, golden dawn

Procedia PDF Downloads 120

Authors: Hsiang-Yu Yuan, Marc Baguelin, Kin O. Kwok, Nimalan Arinaminpathy, Edwin Leeuwen, Steven Riley

Abstract:

Traditional syndromic surveillance for influenza has substantial public health value in characterizing epidemics. Because the relationship between syndromic incidence and the true infection events can vary from one population to another and from one year to another, recent studies rely on combining serological test results with syndromic data from traditional surveillance into epidemic models to make inference on epidemiological processes of influenza. However, despite the widespread availability of serological data, epidemic models have thus far not explicitly represented antibody titre levels and their correspondence with immunity. Most studies use dichotomized data with a threshold (Typically, a titre of 1:40 was used) to define individuals as likely recently infected and likely immune and further estimate the cumulative incidence. Underestimation of Influenza attack rate could be resulted from the dichotomized data. In order to improve the use of serosurveillance data, here, a refinement of the concept of the stratified immunity within an epidemic model for influenza transmission was proposed, such that all individual antibody titre levels were enumerated explicitly and mapped onto a variable scale of susceptibility in different age groups. Haemagglutination inhibition titres from 523 individuals and 465 individuals during pre- and post-pandemic phase of the 2009 pandemic in Hong Kong were collected. The model was fitted to serological data in age-structured population using Bayesian framework and was able to reproduce key features of the epidemics. The effects of age-specific antibody boosting and protection were explored in greater detail. RB was defined to be the effective reproductive number in the presence of stratified immunity and its temporal dynamics was compared to the traditional epidemic model using use dichotomized seropositivity data. Deviance Information Criterion (DIC) was used to measure the fitness of the model to serological data with different mechanisms of the serological response. The results demonstrated that the differential antibody response with age was present (ΔDIC = -7.0). The age-specific mixing patterns with children specific transmissibility, rather than pre-existing immunity, was most likely to explain the high serological attack rates in children and low serological attack rates in elderly (ΔDIC = -38.5). Our results suggested that the disease dynamics and herd immunity of a population could be described more accurately for influenza when the distribution of immunity was explicitly represented, rather than relying only on the dichotomous states 'susceptible' and 'immune' defined by the threshold titre (1:40) (ΔDIC = -11.5). During the outbreak, RB declined slowly from 1.22[1.16-1.28] in the first four months after 1st May. RB dropped rapidly below to 1 during September and October, which was consistent to the observed epidemic peak time in the late September. One of the most important challenges for infectious disease control is to monitor disease transmissibility in real time with statistics such as the effective reproduction number. Once early estimates of antibody boosting and protection are obtained, disease dynamics can be reconstructed, which are valuable for infectious disease prevention and control.

Keywords: effective reproductive number, epidemic model, influenza epidemic dynamics, stratified immunity

Procedia PDF Downloads 232

Authors: J. Gabrielczyk, J. Kluitmann, T. Dammeyer, H. J. Jördening

Abstract:

High-level expression of proteins in bacteria often causes production of insoluble protein aggregates, called inclusion bodies (IB). They contain mainly one type of protein and offer an easy and efficient way to get purified protein. On the other hand, proteins in IB are normally devoid of function and therefore need a special treatment to become active. Most refolding techniques aim at diluting the solubilizing chaotropic agents. Unfortunately, optimal refolding conditions have to be found empirically for every protein. For large-scale applications, a simple refolding process with high yields and high final enzyme concentrations is still missing. The constructed plasmid pASK-IBA63b containing the sequence of fructosyltransferase (FTF, EC 2.4.1.162) from Bacillus subtilis NCIMB 11871 was transformed into E. coli BL21 (DE3) Rosetta. The bacterium was cultivated in a fed-batch bioreactor. The produced FTF was obtained mainly as IB. For refolding experiments, five different amounts of IBs were solubilized in urea buffer with protein concentration of 0.2-8.5 g/L. Solubilizates were refolded with batch or continuous dialysis. The refolding yield was determined by measuring the protein concentration of the clear supernatant before and after the dialysis. Particle size was measured by dynamic light scattering. We tested the solubilization properties of fructosyltransferase IBs. The particle size measurements revealed that the solubilization of the aggregates is achieved at urea concentration of 5M or higher and confirmed by absorption spectroscopy. All results confirm previous investigations that refolding yields are dependent upon initial protein concentration. In batch dialysis, the yields dropped from 67% to 12% and 72% to 19% for continuous dialysis, in relation to initial concentrations from 0.2 to 8.5 g/L. Often used additives such as sucrose and glycerol had no effect on refolding yields. Buffer screening indicated a significant increase in activity but also temperature stability of FTF with citrate/phosphate buffer. By adding citrate to the dialysis buffer, we were able to increase the refolding yields to 82-47% in batch and 90-74% in the continuous process. Further experiments showed that in general, higher ionic strength of buffers had major impact on refolding yields; doubling the buffer concentration increased the yields up to threefold. Finally, we achieved corresponding high refolding yields by reducing the chamber volume by 75% and the amount of buffer needed. The refolded enzyme had an optimal activity of 12.5±0.3 x104 units/g. However, detailed experiments with native FTF revealed a reaggregation of the molecules and loss in specific activity depending on the enzyme concentration and particle size. For that reason, we actually focus on developing a process of simultaneous enzyme refolding and immobilization. The results of this study show a new approach in finding optimal refolding conditions for inclusion bodies at high concentrations. Straightforward buffer screening and increase of the ionic strength can optimize the refolding yield of the target protein by 400%. Gentle removal of chaotrope with continuous dialysis increases the yields by an additional 65%, independent of the refolding buffer applied. In general time is the crucial parameter for successful refolding of solubilized proteins.

Keywords: dialysis, inclusion body, refolding, solubilization

Procedia PDF Downloads 275

Authors: Waqas Ahmed Khan, Faheem Khokhaar

Abstract:

Air pollution is one of the main perpetrators of climate change. GHGs cause melting of glaciers and cause change in temperature and heavy rain fall many gasses like Formaldehyde is not direct precursor that damage ozone like CO2 or Methane but Formaldehyde (HCHO) form glyoxal (CHOCHO) that has effect on ozone. Countries around the globe have unique air quality monitoring protocols to describe local air pollution. Formaldehyde is a colorless, flammable, strong-smelling chemical that is used in building materials and to produce many household products and medical preservatives. Formaldehyde also occurs naturally in the environment. It is produced in small amounts by most living organisms as part of normal metabolic processes. Pakistan lacks the monitoring facilities on larger scale to measure the atmospheric gasses on regular bases. Formaldehyde is formed from Glyoxal and effect mountain biodiversity and livelihood. So its monitoring is necessary in order to maintain and preserve biodiversity. Objective: Present study is aimed to measure atmospheric HCHO vertical column densities (VCDs) obtained from ground-base and compute HCHO data in Punjab and elevated areas (Rawalpindi & Islamabad) by satellite observation during the time period of 2014-2015. Methodology: In order to explore the spatial distributing of H2CO, various fields campaigns including international scientist by using car Max-Doas. Major focus was on the cities along national highways and industrial region of Punjab Pakistan. Level 2 data product of satellite instruments OMI retrieved by differential optical absorption spectroscopy (DOAS) technique are used. Spatio-temporal distribution of HCHO column densities over main cities and region of Pakistan has been discussed. Results: Results show the High HCHO column densities exceeding permissible limit over the main cities of Pakistan particularly the areas with rapid urbanization and enhanced economic growth. The VCDs value over elevated areas of Pakistan like Islamabad, Rawalpindi is around 1.0×1016 to 34.01×1016 Molecules’/cm2. While Punjab has values revolving around the figure 34.01×1016. Similarly areas with major industrial activity showed high amount of HCHO concentrations. Tropospheric glyoxal VCDs were found to be 4.75 × 1015 molecules/cm2. Conclusion: Results shows that monitoring site surrounded by Margalla hills (Islamabad) have higher concentrations of Formaldehyde. Wind data shows that industrial areas and areas having high economic growth have high values as they provide pathways for transmission of HCHO. Results obtained from this study would help EPA, WHO and air protection departments in order to monitor air quality and further preservation and restoration of mountain biodiversity.

Keywords: air quality, formaldehyde, Max-Doas, vertical column densities (VCDs), satellite instrument, climate change

Procedia PDF Downloads 192

Authors: Shagufta Jabeen, Faez J. Firdaus Abdullah, Zunita Zakaria, Nurulfiza M. Isa, Yung C. Tan, Wai Y. Yee, Abdul R. Omar

Abstract:

Pasteurella multocida (PM) is an important veterinary opportunistic pathogen particularly associated with septicemic pasteurellosis, pneumonic pasteurellosis and hemorrhagic septicemia in cattle and buffaloes. P. multocida serotype A has been reported to cause fatal pneumonia and septicemia. Pasteurella multocida subspecies multocida of serotype A Malaysian isolate PMTB2.1 was first isolated from buffaloes died of septicemia. In this study, the genome of P. multocida strain PMTB2.1 was sequenced using third-generation sequencing technology, PacBio RS2 system and analyzed bioinformatically via de novo analysis followed by in-depth analysis based on comparative genomics. Bioinformatics analysis based on de novo assembly of PacBio raw reads generated 3 contigs followed by gap filling of aligned contigs with PCR sequencing, generated a single contiguous circular chromosome with a genomic size of 2,315,138 bp and a GC content of approximately 40.32% (Accession number CP007205). The PMTB2.1 genome comprised of 2,176 protein-coding sequences, 6 rRNA operons and 56 tRNA and 4 ncRNAs sequences. The comparative genome sequence analysis of PMTB2.1 with nine complete genomes which include Actinobacillus pleuropneumoniae, Haemophilus parasuis, Escherichia coli and five P. multocida complete genome sequences including, PM70, PM36950, PMHN06, PM3480, PMHB01 and PMTB2.1 was carried out based on OrthoMCL analysis and Venn diagram. The analysis showed that 282 CDs (13%) are unique to PMTB2.1and 1,125 CDs with orthologs in all. This reflects overall close relationship of these bacteria and supports the classification in the Gamma subdivision of the Proteobacteria. In addition, genomic distance analysis among all nine genomes indicated that PMTB2.1 is closely related with other five Pasteurella species with genomic distance less than 0.13. Synteny analysis shows subtle differences in genetic structures among different P.multocida indicating the dynamics of frequent gene transfer events among different P. multocida strains. However, PM3480 and PM70 exhibited exceptionally large structural variation since they were swine and chicken isolates. Furthermore, genomic structure of PMTB2.1 is more resembling that of PM36950 with a genomic size difference of approximately 34,380 kb (smaller than PM36950) and strain-specific Integrative and Conjugative Elements (ICE) which was found only in PM36950 is absent in PMTB2.1. Meanwhile, two intact prophages sequences of approximately 62 kb were found to be present only in PMTB2.1. One of phage is similar to transposable phage SfMu. The phylogenomic tree was constructed and rooted with E. coli, A. pleuropneumoniae and H. parasuis based on OrthoMCL analysis. The genomes of P. multocida strain PMTB2.1 were clustered with bovine isolates of P. multocida strain PM36950 and PMHB01 and were separated from avian isolate PM70 and swine isolates PM3480 and PMHN06 and are distant from Actinobacillus and Haemophilus. Previous studies based on Single Nucleotide Polymorphism (SNPs) and Multilocus Sequence Typing (MLST) unable to show a clear phylogenetic relatedness between Pasteurella multocida and the different host. In conclusion, this study has provided insight on the genomic structure of PMTB2.1 in terms of potential genes that can function as virulence factors for future study in elucidating the mechanisms behind the ability of the bacteria in causing diseases in susceptible animals.

Keywords: comparative genomics, DNA sequencing, phage, phylogenomics

Procedia PDF Downloads 158

Authors: David Tennison

Abstract:

Epistemic injustice, an injustice done to a person specifically in their capacity as a “knower”, is a subtle form of discrimination, yet its effects can be as dehumanizing and damaging as more overt forms of discrimination. The lens of epistemic injustice has, in recent years, been fruitfully applied to the field of healthcare, examining questions of agency, power, credibility and belief in doctor-patient interactions. Contested illness patients (e.g., those with illnesses lacking scientific consensuses such as fibromyalgia (FM), Myalgic Encephalomyelitis/ Chronic Fatigue Syndrome (ME/CFS) and Long Covid) face higher levels of scrutiny than other patient groups and are often disbelieved or dismissed when their ailments cannot be easily imaged or tested for- often encapsulated by the expression “it’s all in your head”. Using the case study of FM, the trials of contested illness patients in healthcare can be conceptualized in terms of epistemic injustice, and what is going wrong in these doctor-patient relationships can be effectively diagnosed. This case study also helps reveal epistemic dysfunction (structural epistemic issues embedded in the healthcare system), how this relates to stigma identity-based prejudice, and how the healthcare system upholds existing societal hierarchies and disenfranchises the most vulnerable. In the modern landscape, where cases of these chronic illnesses are not only on the rise but future pandemics threaten to add to their number, this conversation is crucial for the well-being of patients and providers. This presentation will cover what epistemic injustice is and how it can be applied to the politics of the doctor-patient interaction on a micro level and the politics of the healthcare system more broadly. Contested illnesses will be explored in terms of how the “contested” label causes the patient to experience disease stigma and lowers their credibility in healthcare and across other aspects of life. This will be explored in tandem with a discussion of existing identity-based prejudice in the healthcare system and how social identities (such as those of gender, race, and socioeconomic status) intersect with the contested illness label. The effects of epistemic injustice, which include worsening patients’ symptoms of mental health and potentially disenfranchising them from the healthcare system altogether, will be presented alongside the potential ethical quandaries this poses for providers. Finally, issues with the way healthcare appointments and the modern NHS function will be explored in terms of epistemic injustice and solutions to improve doctor-patient communication and patient care will be discussed. The relationship between contested illness patients and healthcare providers is notoriously poor, and while this can mean frustration or feelings of unfulfillment in providers, the negative effects for patients are much more severe. The purpose of this research, then, is to highlight these issues and suggest ways in which to improve the healthcare experience for these patients, along with improving doctor-patient communication and mending the doctor-patient relationship in a tangible and realistic way. This research also aims to provoke important conversations about belief and hierarchy in medical settings and how these aspects intersect with identity prejudices.

Keywords: epistemic injustice, fibromyalgia, contested illnesses, chronic illnesses, doctor-patient relationships, philosophy of medicine

Procedia PDF Downloads 37

Authors: Quan Wen, Tianxiao Chang, Shaolu Shi, Yushi Wang, Guangyu Wang

Abstract:

As a kind of working environment of the fuze, the spin rate decaying law of projectile in exterior trajectory is of great value in the design of the rotation count fixed distance fuze. In addition, it is significant in the field of devices for simulation tests of fuze exterior ballistic environment, flight stability, and dispersion accuracy of gun projectile and opening and scattering design of submunition and illuminating cartridges. Besides, the self-destroying mechanism of the fuze in small-caliber projectile often works by utilizing the attenuation of centrifugal force. In the theory of projectile aerodynamics and fuze design, there are many formulas describing the change law of projectile angular velocity in external ballistic such as Roggla formula, exponential function formula, and power function formula. However, these formulas are mostly semi-empirical due to the poor test conditions and insufficient test data at that time. These formulas are difficult to meet the design requirements of modern fuze because they are not accurate enough and have a narrow range of applications now. In order to provide more accurate ballistic environment parameters for the design of a hemispherical head projectile fuze, the projectile’s spin rate decaying law in exterior trajectory under the effect of air resistance was studied. In the analysis, the projectile shape was simplified as hemisphere head, cylindrical part, rotating band part, and anti-truncated conical tail. The main assumptions are as follows: a) The shape and mass are symmetrical about the longitudinal axis, b) There is a smooth transition between the ball hea, c) The air flow on the outer surface is set as a flat plate flow with the same area as the expanded outer surface of the projectile, and the boundary layer is turbulent, d) The polar damping moment attributed to the wrench hole and rifling mark on the projectile is not considered, e) The groove of the rifle on the rotating band is uniform, smooth and regular. The impacts of the four parts on aerodynamic moment of the projectile rotation were obtained by aerodynamic theory. The surface friction stress of the projectile, the polar damping moment formed by the head of the projectile, the surface friction moment formed by the cylindrical part, the rotating band, and the anti-truncated conical tail were obtained by mathematical derivation. After that, the mathematical model of angular spin rate attenuation was established. In the whole trajectory with the maximum range angle (38°), the absolute error of the polar damping torque coefficient obtained by simulation and the coefficient calculated by the mathematical model established in this paper is not more than 7%. Therefore, the credibility of the mathematical model was verified. The mathematical model can be described as a first-order nonlinear differential equation, which has no analytical solution. The solution can be only gained as a numerical solution by connecting the model with projectile mass motion equations in exterior ballistics.

Keywords: ammunition engineering, fuze technology, spin rate, numerical simulation

Procedia PDF Downloads 113

Authors: Margaret Boone Rappaport, Christopher J. Corbally

Abstract:

The ancient ape ancestral population from which living great ape and human species evolved had demographic features affecting their evolution. The population was large, had great genetic variability, and natural selection was effective at honing adaptations. The emerging populations of chimpanzees and humans were affected more by founder effects and genetic drift because they were smaller. Natural selection did not disappear, but it was not as strong. Consequences of the 'population crash' and the human effective population size are introduced briefly. The history of the ancient apes is written in the genomes of living humans and great apes. The expansion of the brain began before the human line emerged. Coalescence times for some genes are very old – up to several million years, long before Homo sapiens. The mismatch between gene trees and species trees highlights the anthropoid speciation processes, and gives the human genome history a fuzzy, probabilistic quality. However, it suggests traits that might form a foundation for capacities emerging later. A theoretical model is presented in which the genomes of early ape populations provide the substructure for the emergence of religious capacity later on the human line. The model does not search for religion, but its foundations. It suggests a course by which an evolutionary line that began with prosimians eventually produced a human species with biologically based religious capacity. The model of the sequential emergence of religious capacity relies on cognitive science, neuroscience, paleoneurology, primate field studies, cognitive archaeology, genomics, and population genetics. And, it emphasizes five trait types: (1) Documented, positive selection of sensory capabilities on the human line may have favored survival, but also eventually enriched human religious experience. (2) The bonobo model suggests a possible down-regulation of aggression and increase in tolerance while feeding, as well as paedomorphism – but, in a human species that remains cognitively sharp (unlike the bonobo). The two species emerged from the same ancient ape population, so it is logical to search for shared traits. (3) An up-regulation of emotional sensitivity and compassion seems to have occurred on the human line. This finds support in modern genetic studies. (4) The authors’ published model of morality's emergence in Homo erectus encompasses a cognitively based, decision-making capacity that was hypothetically overtaken, in part, by religious capacity. Together, they produced a strong, variable, biocultural capability to support human sociability. (5) The full flowering of human religious capacity came with the parietal expansion and smaller face (klinorhynchy) found only in Homo sapiens. Details from paleoneurology suggest the stage was set for human theologies. Larger parietal lobes allowed humans to imagine inner spaces, processes, and beings, and, with the frontal lobe, led to the first theologies composed of structured and integrated theories of the relationships between humans and the supernatural. The model leads to the evolution of a small population of African hominins that was ready to emerge with religious capacity when the species Homo sapiens evolved two hundred thousand years ago. By 50-60,000 years ago, when human ancestors left Africa, they were fully enabled.

Keywords: genetic drift, genomics, parietal expansion, religious capacity

Procedia PDF Downloads 317

Authors: Deborah A. Hollingworth

Abstract:

The key tenets associated with ‘environmental justice’, were first articulated in a global context in Principle 10 of the United Nations Declaration on Environment and Development at Rio de Janeiro in 1992 (the Rio Declaration). The elements can be conflated to require: public participation in decision-making; the provision of relevant information to those affected about environmental hazards issues; access to judicial and administrative proceeding; and the opportunity for redress where remedy where required. This paper examines the legislative and regulatory arrangements in place for the implementation these elements in a number of industrialised democracies, including Australia. Most have, over time made regulatory provision for these elements – even if they are not directly attributed Principle 10 or the notion of environmental justice. The paper proposes, that of these elements the most critical to the achievement of good environmental governance, is a legislated recognition and role of public participation. However, the paper considers that notwithstanding sound legislative and regulatory practices, environmental regulators frequently struggle, where there is a complex decision-making scenario or long-standing enmity between a community and industry to achieve effective engagement with the public. This study considers the dilemma confronted by environmental regulators to given meaningful effect to the principles enshrined in Principle 10 – that even when the legislative expression of Principle 10 is adhered to – does not prevent adverse outcomes. In particular, it considers, as a case study a prominent environmental incident in 2014 in Australia in which an open-cut coalmine located in the regional township of Morwell caught fire during bushfire season. The fire, which took 45 days to be extinguished had a significant and adverse impact on the community in question, but compounded a complex, and sometime antagonistic history between the mine and township. The case study exemplifies the complex factors that will often be present between industry, the public and regulatory bodies, and which confound the concept of environmental justice, and the elements of enshrined in the Principle 10 of the Rio Declaration. The study proposes that such tensions and complex examples will commonly be the reality of communities and regulators. However, to give practical effect to outcomes contemplated by Principle 10, the paper considers that regulators will may consider public intervention more broadly as including early interventions and formal opportunities for “conferencing” between industry, community and regulators. These initiatives help to develop a shared understanding and identification of issues. It is proposed that although important, options for “alternative dispute resolution” are not sufficiently preventative, as they come into play when a dispute has arise. Similarly “restorative justice” programs, while important once an incident or adverse environmental outcome has occurred, are post event and therefore necessarily limited. The paper considers the examples of how public participation at the outset – at the time of a proposal, before issues arise or eventuate to ensure, is demonstrably the most effective way for building commonality and an agreed methodology for working to resolve issues once they occur.

Keywords: environmental justice, alternative dispute resolution, domestic environmental law, international environmental law

Procedia PDF Downloads 277

Authors: Thomas James Bell III

Abstract:

Pedagogy is thought of as one's philosophy, theory, or teaching method. This study examines the science of learning, considering the forced reconsideration of effective pedagogy brought on by the aftermath of the 2020 coronavirus pandemic. With the aid of various technologies, online education holds challenges and promises to enhance the learning environment if implemented to facilitate student learning. Behaviorism centers around the belief that the instructor is the sage on the classroom stage using repetition techniques as the primary learning instrument. This approach to pedagogy ascribes complete control of the learning environment and works best for students to learn by allowing students to answer questions with immediate feedback. Such structured learning reinforcement tends to guide students' learning without considering learners' independence and individual reasoning. And such activities may inadvertently stifle the student's ability to develop critical thinking and self-expression skills. Fundamentally liberationism pedagogy dismisses the concept that education is merely about students learning things and more about the way students learn. Alternatively, the liberationist approach democratizes the classroom by redefining the role of the teacher and student. The teacher is no longer viewed as the sage on the stage but as a guide on the side. Instead, this approach views students as creators of knowledge and not empty vessels to be filled with knowledge. Moreover, students are well suited to decide how best to learn and which areas improvements are needed. This study will explore the classroom instructor as a servant leader in the twenty-first century, which allows students to integrate technology that encapsulates more individual learning styles. The researcher will examine the Professional Scrum Master (PSM I) exam pass rate results of 124 students in six sections of an Agile scrum course. The students will be separated into two groups; the first group will follow a structured instructor-led course outlined by a course syllabus. The second group will consist of several small teams (ten or fewer) of self-led and self-empowered students. The teams will conduct several event meetings that include sprint planning meetings, daily scrums, sprint reviews, and retrospective meetings throughout the semester will the instructor facilitating the teams' activities as needed. The methodology for this study will use the compare means t-test to compare the mean of an exam pass rate in one group to the mean of the second group. A one-tailed test (i.e., less than or greater than) will be used with the null hypothesis, for the difference between the groups in the population will be set to zero. The major findings will expand the pedagogical approach that suggests pedagogy primarily exist in support of teacher-led learning, which has formed the pillars of traditional classroom teaching. But in light of the fourth industrial revolution, there is a fusion of learning platforms across the digital, physical, and biological worlds with disruptive technological advancements in areas such as the Internet of Things (IoT), artificial intelligence (AI), 3D printing, robotics, and others.

Keywords: pedagogy, behaviorism, liberationism, flipping the classroom, servant leader instructor, agile scrum in education

Procedia PDF Downloads 110

Authors: Magdalena Chrabąszcz, Teresa Wolniewicz, Cezary Maciejewski, Joanna Czuwara

Abstract:

BACKGROUND: Dermoscopy is a clinical tool known to improve the early detection of melanoma and other malignancies of the skin. Over the past few years melanoma has grown into a disease of socio-economic importance due to the increasing incidence and persistently high mortality rates. Early diagnosis remains the best method to reduce melanoma and non-melanoma skin cancer– related mortality and morbidity. Dermoscopy is a noninvasive technique that consists of viewing pigmented skin lesions through a hand-held lens. This simple procedure increases melanoma diagnostic accuracy by up to 35%. Dermoscopy is currently the standard for clinical differential diagnosis of cutaneous melanoma and for qualifying lesion for the excision biopsy. Like any clinical tool, training is required for effective use. The introduction of small and handy dermoscopes contributed significantly to the switch of dermatoscopy toward a first-level useful tool. Non-dermatologist physicians are well positioned for opportunistic melanoma detection; however, education in the skin cancer examination is limited during medical school and traditionally lecture-based. AIM: The aim of this randomized study was to determine whether the adjunct of dermoscopy to the standard fourth year medical curriculum improves the ability of medical students to distinguish between benign and malignant lesions and assess acceptability and satisfaction with the intervention. METHODS: We performed a prospective study in 2 cohorts of fourth-year medical students at Medical University of Warsaw. Groups having dermatology course, were randomly assigned to:  cohort A: with limited access to dermatoscopy from their teacher only – 1 dermatoscope for 15 people  Cohort B: with a full access to use dermatoscopy during their clinical classes:1 dermatoscope for 4 people available constantly plus 15-minute dermoscopy tutorial. Students in both study arms got an image-based test of 10 lesions to assess ability to differentiate benign from malignant lesions and postintervention survey collecting minimal background information, attitudes about the skin cancer examination and course satisfaction. RESULTS: The cohort B had higher scores than the cohort A in recognition of nonmelanocytic (P < 0.05) and melanocytic (P <0.05) lesions. Medical students who have a possibility to use dermatoscope by themselves have also a higher satisfaction rates after the dermatology course than the group with limited access to this diagnostic tool. Moreover according to our results they were more motivated to learn dermatoscopy and use it in their future everyday clinical practice. LIMITATIONS: There were limited participants. Further study of the application on clinical practice is still needed. CONCLUSION: Although the use of dermatoscope in dermatology as a specialty is widely accepted, sufficiently validated clinical tools for the examination of potentially malignant skin lesions are lacking in general practice. Introducing medical students to dermoscopy in their fourth year curricula of medical school may improve their ability to differentiate benign from malignant lesions. It can can also encourage students to use dermatoscopy in their future practice which can significantly improve early recognition of malignant lesions and thus decrease melanoma mortality.

Keywords: dermatoscopy, early detection of melanoma, medical education, skin cancer

Procedia PDF Downloads 96

Authors: Andreas Latz, Daniela Heinz, Fatima Hashemi, Melek Baygül

Abstract:

Introduction: Feline leukemia virus (FeLV), feline immunodeficiency virus (FIV), and feline coronavirus (FCoV) are serious infectious diseases affecting cats worldwide. Transmission of these viruses occurs primarily through close contact with infected cats (via saliva, nasal secretions, faeces, etc.). FeLV, FIV, and FCoV infections can occur in combination and are expressed in similar clinical symptoms. Diagnosis can therefore be challenging: Symptoms are variable and often non-specific. Sick cats show very similar clinical symptoms: apathy, anorexia, fever, immunodeficiency syndrome, anemia, etc. Sample volume for small companion animals for diagnostic purposes can be challenging to collect. In addition, multiplex diagnosis of diseases can contribute to an easier, cheaper, and faster workflow in the lab as well as to the better differential diagnosis of diseases. For this reason, we wanted to develop a new diagnostic tool that utilizes less sample volume, reagents, and consumables than multiplesingleplex ELISA assays Methods: The Multiplier from Dynextechonogies (USA) has been used as platform to develop a Multiplex diagnostic tool for the detection of antibodies against FIV and FCoV/FIP and antigens for FeLV. Multiplex diagnostics. The Dynex®Multiplier®is a fully automated chemiluminescence immunoassay analyzer that significantly simplifies laboratory workflow. The Multiplier®ease-of-use reduces pre-analytical steps by combining the power of efficiently multiplexing multiple assays with the simplicity of automated microplate processing. Plastic beads have been coated with antigens for FIV and FCoV/FIP, as well as antibodies for FeLV. Feline blood samples are incubated with the beads. Read out of results is performed via chemiluminescence Results: Bead coating was optimized for each individual antigen or capture antibody and then combined in the multiplex diagnostic tool. HRP: Antibody conjugates for FIV and FCoV antibodies, as well as detection antibodies for FeLV antigen, have been adjusted and mixed. 3 individual prototyple batches of the assay have been produced. We analyzed for each disease 50 well defined positive and negative samples. Results show an excellent diagnostic performance of the simultaneous detection of antibodies or antigens against these feline diseases in a fully automated system. A 100% concordance with singleplex methods like ELISA or IFA can be observed. Intra- and Inter-Assays showed a high precision of the test with CV values below 10% for each individual bead. Accelerated stability testing indicate a shelf life of at least 1 year. Conclusion: The new tool can be used for multiplex diagnostics of the most important feline infectious diseases. Only a very small sample volume is required. Fully automation results in a very convenient and fast method for diagnosing animal diseases.With its large specimen capacity to process over 576 samples per 8-hours shift and provide up to 3,456 results, very high laboratory productivity and reagent savings can be achieved.

Keywords: Multiplex, FIV, FeLV, FCoV, FIP

Procedia PDF Downloads 76

Authors: Hend Ben Tkhayat , Khaled Al Zahabi, Husam Younes

Abstract:

Introduction: Vildagliptin (VG), a dipeptidyl peptidase-4 inhibitor (DPP-4), was proven to be an active agent for the treatment of type 2 diabetes. VG works by enhancing and prolonging the activity of incretins which improves insulin secretion and decreases glucagon release, therefore lowering blood glucose level. It is usually used with various classes, such as insulin sensitizers or metformin. VG is currently only marketed as an immediate-release tablet that is administered twice daily. In this project, we aim to formulate an extended-release with a zero-order profile tableted lipid microparticles of VG that could be administered once daily ensuring the patient’s convenience. Method: The spray-congealing technique was used to prepare VG microparticles. Compritol® was heated at 10 oC above its melting point and VG was dispersed in the molten carrier using a homogenizer (IKA T25- USA) set at 13000 rpm. VG dispersed in the molten Compritol® was added dropwise to the molten Gelucire® 50/13 and PEG® (400, 6000, and 35000) in different ratios under manual stirring. The molten mixture was homogenized and Carbomer® amount was added. The melt was pumped through the two-fluid nozzle of the Buchi® Spray-Congealer (Buchi B-290, Switzerland) using a Pump drive (Master flex, USA) connected to a silicone tubing wrapped with silicone heating tape heated at the same temperature of the pumped mix. The physicochemical properties of the produced VG-loaded microparticles were characterized using Mastersizer, Scanning Electron Microscope (SEM), Differential Scanning Calorimeter (DSC) and X‐Ray Diffractometer (XRD). VG microparticles were then pressed into tablets using a single punch tablet machine (YDP-12, Minhua pharmaceutical Co. China) and in vitro dissolution study was investigated using Agilent Dissolution Tester (Agilent, USA). The dissolution test was carried out at 37±0.5 °C for 24 hours in three different dissolution media and time phases. The quantitative analysis of VG in samples was realized using a validated High-Pressure Liquid Chromatography (HPLC-UV) method. Results: The microparticles were spherical in shape with narrow distribution and smooth surface. DSC and XRD analyses confirmed the crystallinity of VG that was lost after being incorporated into the amorphous polymers. The total yields of the different formulas were between 70% and 80%. The VG content in the microparticles was found to be between 99% and 106%. The in vitro dissolution study showed that VG was released from the tableted particles in a controlled fashion. The adjustment of the hydrophilic/hydrophobic ratio of excipients, their concentration and the molecular weight of the used carriers resulted in tablets with zero-order kinetics. The Gelucire 50/13®, a hydrophilic polymer was characterized by a time-dependent profile with an important burst effect that was decreased by adding Compritol® as a lipophilic carrier to retard the release of VG which is highly soluble in water. PEG® (400,6000 and 35 000) were used for their gelling effect that led to a constant rate delivery and achieving a zero-order profile. Conclusion: Tableted spray-congealed lipid microparticles for extended-release of VG were successfully prepared and a zero-order profile was achieved.

Keywords: vildagliptin, spray congealing, microparticles, controlled release

Procedia PDF Downloads 103