Search results for: native corpus

Authors: Siddhant Sethi, Yasuharu Takashima, Shigetaka Nakamura, Kenzo Fujimoto

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Site-directed mutagenesis is a renowned technique to introduce specific mutations in the genome. To achieve site-directed mutagenesis, many chemical and enzymatic approaches have been reported in the past like disulphite induced genome editing, CRISPR-Cas9, TALEN etc. The chemical methods are invasive whereas the enzymatic approaches are time-consuming and expensive. Most of these techniques are unusable in the cellular application due to their toxicity and other limitations. Photo-chemical cytosine deamination, introduced in 2010, is one of the major technique for enzyme-free single-point mutation of cytosine to uracil in DNA and RNA, wherein, 3-cyanovinylcarbazole nucleoside (CNVK) containing oligodeoxyribonucleotide (ODN) having CNVK at -1 position to that of target cytosine is reversibly crosslinked to target DNA strand using 366 nm and then incubated at 90ºC to accommodate deamination. This technique is superior to enzymatic methods of site-directed mutagenesis but has a disadvantage that it requires the use of high temperature for the deamination step which restricts its applicability in the in vivo applications. This study has been focused on improving the technique by reducing the temperature required for deamination. Firstly, the photo-cross-linker, CNVK has been modified by replacing cyano group attached to vinyl group with methyl ester (OMeVK), amide (NH2VK), and carboxylic acid (OHVK) to observe the acceleration in the deamination of target cytosine cross-linked to vinylcarbazole derivative. Among the derivatives, OHVK has shown 2 times acceleration in deamination reaction as compared to CNVK, while the other two derivatives have shown deceleration towards deamination reaction. The trend of rate of deamination reaction follows the same order as that of hydrophilicity of the vinylcarbazole derivatives. OHVK being most hydrophilic has shown highest acceleration while OMeVK is least hydrophilic has proven to be least active for deamination. Secondly, in the related study, the counter-base of the target cytosine, guanine has been replaced by inosine, 2-aminopurine, nebularine, and 5-nitroindole having distinct hydrogen bonding patterns with target cytosine. Among the ODNs with these counter bases, ODN with inosine has shown 12 fold acceleration towards deamination of cytosine cross-linked to CNVK at physiological conditions as compared to guanosine. Whereas, when 2-aminopurine, nebularine, and 5-nitroindole were used, no deamination reaction took place. It can be concluded that inosine has potential to be used as the counter base of target cytosine for the CNVK mediated photo-cross-linking induced deamination of cytosine. The increase in rate of deamination reaction has been attributed to pattern and number of hydrogen bonding between the cytosine and counter base. One of the important factor is presence of hydrogen bond between exo-cyclic amino group of cytosine and the counter base. These results will be useful for development of more efficient technique for site-directed mutagenesis for C → U transformations in the DNA/RNA which might be used in the living system for treatment of various genetic disorders and genome engineering for making designer and non-native proteins.

Keywords: C to U transformation, DNA editing, genome engineering, ultra-fast photo-cross-linking

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Authors: L. J. de Jager

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In the South African English school curriculum, one of the aims is to achieve communicative competence, the knowledge of using language competently and appropriately in a speech community. Communicatively competent speakers should not only produce grammatically correct sentences but also produce contextually appropriate sentences for various purposes and in different situations. As most speakers of English are non-native speakers, achieving communicative competence remains a complex challenge. Moreover, the changing needs of society necessitate not merely language proficiency, but also technological proficiency. One of the burning issues in the South African educational landscape is the replacement of the standardised literacy model by the pedagogy of multiliteracies that incorporate, by default, the exploration of technological text forms that are part of learners’ everyday lives. It foresees learners as decoders, encoders, and manufacturers of their own futures by exploiting technological possibilities to constantly create and recreate meaning. As such, 21st century learners will feel comfortable working with multimodal texts that are intrinsically part of their lives and by doing so, become authors of their own learning experiences while teachers may become agents supporting learners to discover their capacity to acquire new digital skills for the century of multiliteracies. The aim is transformed practice where learners use their skills, ideas, and knowledge in new contexts. This paper reports on a research project on the integration of technology for language learning, based on the technological pedagogical content knowledge framework, conceptually founded in the theory of multiliteracies, and which aims to achieve communicative competence. The qualitative study uses the community of inquiry framework to answer the research question: How does the integration of technology transform language teaching of preservice teachers? Pre-service teachers in the Postgraduate Certificate of Education Programme with English as methodology were purposively selected to source and evaluate apps for teaching and learning English. The participants collaborated online in a dedicated Blackboard module, using discussion threads to sift through applicable apps and develop interactive lessons using the Apps. The selected apps were entered on to a predesigned Qualtrics form. Data from the online discussions, focus group interviews, and reflective journals were thematically and inductively analysed to determine the participants’ perceptions and experiences when integrating technology in lesson design and the extent to which communicative competence was achieved when using these apps. Findings indicate transformed practice among participants and research team members alike with a better than average technology acceptance and integration. Participants found value in online collaboration to develop and improve their own teaching practice by experiencing directly the benefits of integrating e-learning into the teaching of languages. It could not, however, be clearly determined whether communicative competence was improved. The findings of the project may potentially inform future e-learning activities, thus supporting student learning and development in follow-up cycles of the project.

Keywords: apps, communicative competence, English teaching, technology integration, technological pedagogical content knowledge

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Authors: Mohammed Sabbah, Prospero Di Pierro, Raffaele Porta

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Protein-based edible films and coatings have attracted an increasing interest in recent years since they might be used to protect pharmaceuticals or improve the shelf life of different food products. Among them, several plant proteins represent an abundant, inexpensive and renewable raw source. These natural biopolymers are used as film forming agents, being able to form intermolecular linkages by various interactions. However, without the addition of a plasticizing agent, many biomaterials are brittle and, consequently, very difficult to be manipulated. Plasticizers are generally small and non-volatile organic additives used to increase film extensibility and reduce its crystallinity, brittleness and water vapor permeability. Plasticizers normally act by decreasing the intermolecular forces along the polymer chains, thus reducing the relative number of polymer-polymer contacts, producing a decrease in cohesion and tensile strength and thereby increasing film flexibility allowing its deformation without rupture. The most commonly studied plasticizers are polyols, like glycerol (GLY) and some mono or oligosaccharides. In particular, GLY not only increases film extensibility but also migrates inside the film network often causing the loss of desirable mechanical properties of the material. Therefore, replacing GLY with a different plasticizer might help to improve film characteristics allowing potential industrial applications. To improve film properties, it seemed of interest to test as plasticizers some cationic small molecules like polyamines (PAs). Putrescine, spermidine (SPD), and spermine are PAs widely distributed in nature and of particular interest for their biological activities that may have some beneficial health effects. Since PAs contains amino instead of hydroxyl functional groups, they are able to trigger ionic interactions with negatively charged proteins. Bitter vetch (Vicia ervilia; BV) is an ancient grain legume crop, originated in the Mediterranean region, which can be found today in many countries around the world. This annual Vicia genus shows several favorable features, being their seeds a cheap and abundant protein source. The main objectives of this study were to investigate the effect of different concentrations of SPD on the mechanical and permeability properties of films prepared with native or heat denatured BV proteins in the presence of different concentrations of SPD and/or GLY. Therefore, a BV seed protein concentrate (BVPC), containing about 77% proteins, was used to prepare film forming solutions (FFSs), whereas GLY and SPD were added as film plasticizers, either singly or in combination, at various concentrations. Since a primary plasticizer is generally defined as a molecule that when added to a material makes it softer, more flexible and easier to be processed, our findings lead to consider SPD as a possible primary plasticizer of protein-based films. In fact, the addition of millimolar concentrations of SPD to BVPC FFS allowed obtaining handleable biomaterials with improved properties. Moreover, SPD can be also considered as a secondary plasticizer, namely an 'extender', because of its ability even to enhance the plasticizing performance of GLY. In conclusion, our studies indicate that innovative edible protein-based films and coatings can be obtained by using PAs as new plasticizers.

Keywords: edible films, glycerol, plasticizers, polyamines, spermidine

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Authors: Soledad Coronel Poma, Lorena Alvarado Rodriguez

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The purpose of this project is to design public space through urban-architectural strategies that help to the sustainable recovery of the Huayla estuary and the revival of tourism in this area. This design considers other sustainable and architectural ideas used in similar cases, along with national and international regulations for saving shorelines in danger. To understand the situation of this location, Puerto Bolivar is the main port of the Province of El Oro and of the south of the country, where 90,000 national and foreign tourists pass through all year round. For that reason, a physical-urban, social, and environmental analysis of the area was carried out through surveys and conversations with the community. This analysis showed that around 70% of people feel unsatisfied and concerned about the estuary and its surroundings. Crime, absence of green areas, bad conservation of shorelines, lack of tourists, poor commercial infrastructure, and the spread of informal commerce are the main issues to be solved. As an intervention project whose main goal is that residents and tourists have contact with native nature and enjoy doing local activities, three main strategies: mobility, ecology, and urban –architectural are proposed to recover the estuary and its surroundings. First of all, the design of this public space is based on turning the estuary location into a linear promenade that could be seen as a tourist corridor, which would help to reduce pollution, increase green spaces and improve tourism. Another strategy aims to improve the economy of the community through some local activities like fishing and sailing and the commerce of fresh seafood, both raw products and in restaurants. Furthermore, in support of the environmental approach, some houses are rebuilt as sustainable houses using local materials and rearranged into blocks closer to the commercial area. Finally, the planning incorporates the use of many plants such as palms, sameness trees, and mangroves around the area to encourage people to get in touch with nature. The results of designing this space showed an increase in the green area per inhabitant index. It went from 1.69 m²/room to 10.48 m²/room, with 12 096 m² of green corridors and the incorporation of 5000 m² of mangroves at the shoreline. Additionally, living zones also increased with the creation of green areas taking advantage of the existing nature and implementing restaurants and recreational spaces. Moreover, the relocation of houses and buildings helped to free estuary's shoreline, so people are now in more comfortable places closer to their workplaces. Finally, dock spaces are increased, reaching the capacity of the boats and canoes, helping to organize the area in the estuary. To sum up, this project searches the improvement of the estuary environment with its shoreline and surroundings that include the vegetation, infrastructure and people with their local activities, achieving a better quality of life, attraction of tourism, reduction of pollution and finally getting a full recovered estuary as a natural ecosystem.

Keywords: recover, public space, stuary, sustainable

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Authors: G. M. Vlasceanu, H. Iovu, E. Vasile, M. Ionita

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Herein, the synthesis and characterization of chitosan-gelatin highly porous scaffold reinforced with graphene oxide, and hydroxyapatite (HAp), crosslinked with genipin was targeted. In tissue engineering, chitosan and gelatin are two of the most robust biopolymers with wide applicability due to intrinsic biocompatibility, biodegradability, low antigenicity properties, affordability, and ease of processing. HAp, per its exceptional activity in tuning cell-matrix interactions, is acknowledged for its capability of sustaining cellular proliferation by promoting bone-like native micro-media for cell adjustment. Genipin is regarded as a top class cross-linker, while graphene oxide (GO) is viewed as one of the most performant and versatile fillers. The composites with natural bone HAp/biopolymer ratio were obtained by cascading sonochemical treatments, followed by uncomplicated casting methods and by freeze-drying. Their structure was characterized by Fourier Transform Infrared Spectroscopy and X-ray Diffraction, while overall morphology was investigated by Scanning Electron Microscopy (SEM) and micro-Computer Tomography (µ-CT). Ensuing that, in vitro enzyme degradation was performed to detect the most promising compositions for the development of in vivo assays. Suitable GO dispersion was ascertained within the biopolymer mix as nanolayers specific signals lack in both FTIR and XRD spectra, and the specific spectral features of the polymers persisted with GO load enhancement. Overall, correlations between the GO induced material structuration, crystallinity variations, and chemical interaction of the compounds can be correlated with the physical features and bioactivity of each composite formulation. Moreover, the HAp distribution within follows an auspicious density gradient tuned for hybrid osseous/cartilage matter architectures, which were mirrored in the mice model tests. Hence, the synthesis route of a natural polymer blend/hydroxyapatite-graphene oxide composite material is anticipated to emerge as influential formulation in bone tissue engineering. Acknowledgement: This work was supported by the project 'Work-based learning systems using entrepreneurship grants for doctoral and post-doctoral students' (Sisteme de invatare bazate pe munca prin burse antreprenor pentru doctoranzi si postdoctoranzi) - SIMBA, SMIS code 124705 and by a grant of the National Authority for Scientific Research and Innovation, Operational Program Competitiveness Axis 1 - Section E, Program co-financed from European Regional Development Fund 'Investments for your future' under the project number 154/25.11.2016, P_37_221/2015. The nano-CT experiments were possible due to European Regional Development Fund through Competitiveness Operational Program 2014-2020, Priority axis 1, ID P_36_611, MySMIS code 107066, INOVABIOMED.

Keywords: biopolymer blend, ectopic osteoinduction, graphene oxide composite, hydroxyapatite

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Authors: Aathira Peedikaparambil Somasundaran

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The beginning of the 20th century marked the perios when a new generation of Lebanese radicals sowed the seeds for the second phase of Levant literary modernism, situated in the Levant. Beirut, during this era popularly fit every radical writer’s criterion owing to its weakened censorship and political control, despite the absence of a protective womb for the development of literary modernism, caused by the natively prevalent political unsettlement. The third stage of literary modernization, in which scholars used Western-inspired critical techniques to better understand their own cultures, coincides with the time period examined in this paper, which involved the international-inspired critical analysis of native cultural stimulants, which raised questions among Arab freethinking intellectuals. Locals who ventured outside recognised the difference between the West's progress and their own nations' stagnation. The awareness of such ‘gap of success’ aroused an ambition from journalists, authors, and proletarian revolutionaries who had studied in Europe, and finally developed enlightened ideas. Some Middle Eastern authors and artists only adopted current social and political frameworks after discovering western modernity. After learning about the upheavals that were happening in the West, these thinkers aspired to bring about equally broad drastic developments in their own country's social, political, and cultural milieu. These occurrences illustrate the increased power of migration to alter the cultural and literary scene in the Levant. The paper intends to discuss the different effects of migration that contributed to Levant literary modernism. The exploration of these factors as causes begins with addressing the politically influenced activism, that has always been a relevant part of Beirut, and then diving into the psychological effects of migration in the individuals of the society, which might have induced an accommodability to alien thoughts and ideas over time, as a coping mechanism. Nature or environmental stimuli, a common trigger for any creative output, often having the highest influence during travel will be identified and analysed to inspect the extent of its impact on the exchange of ideas that resulted in Levant modernism. The efficiency of both the stimulating component of travel and the diaspora of the indigenous, a by-product of travel in catalysing modernism in the Levant has to be proven in order to understand how migration indirectly affected the transmission and adoption of ideas in Levant literature. The paper will revisit the events revolving around these key players and platforms like Shir, to understand how the Lebanese literature, tied down in poetry drastically mutated under the leadership of Adonis, Yusuf et Khal, and other pioneers of Levant literary modernism. The conclision will identify the triggers that helped authors overcome personal and geographical barriers to unite the West and the Levant, and investigate the extent to which the bi-directional migration prompted a transformation in the local poetry. Consequently, the paper aims to shed light into the unique factor that provoked the shift in the literary scene of Twentieth century in the Middle East.

Keywords: literature, modernism, Middle East, levant, Beirut

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Authors: Rodrigo M. Falleiro, Joaquim P. L. Parime, Luciano C. Santos, Rodrigo D. Silva

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The Cerrado biome is the most biodiverse savanna on the planet. Located in central Brazil, its preservation is seriously threatened by the advance of intensive agriculture and livestock. Conservation Units and Indigenous Lands are increasingly isolated and subject to mega wildfires. Among the characteristics of this savanna, we highlight the high rate of primary biomass production and the reduced occurrence of large grazing animals. In this biome, the predominant fauna is more dependent on the fruits produced by the dicotyledonous species in relation to other tropical savannas. Fire is a key element in the balance between mono and dicotyledons or between the arboreal and herbaceous strata. Therefore, applying fire regimes that maintain the balance between these strata without harming fruit production is essential in the conservation strategies of Cerrado's biodiversity. Recently, Integrated Fire Management has started to be implemented in Brazilian protected areas. As a result, management with prescribed burns has increasingly replaced strategies based on fire exclusion, which in practice have resulted in large wildfires, with highly negative impacts on fruit and fauna production. In the Indigenous Lands, these fires were carried out respecting traditional knowledge. The indigenous people showed great concern about the effects of fire on fruit plants and important animals. They recommended that the burns be carried out between April and May, as it would result in a greater production of edible fruits ("fruiting burning"). In other tropical savannas in the southern hemisphere, the preferential period tends to be later, in the middle of the dry season, when the grasses are dormant (June to August). However, in the Cerrado, this late period coincides with the flowering and sprouting of several important fruit species. To verify the best burning season, the present work evaluated the effects of fire on flowering and fruit production of theByrsonima sp., Mouriri pusa, Caryocar brasiliense, Anacardium occidentale, Pouteria ramiflora, Hancornia speciosa, Byrsonima verbascifolia, Anacardium humille and Talisia subalbens. The evaluations were carried out in the field, covering 31 Indigenous Lands that cover 104,241.18 Km², where 3,386 prescribed burns were carried out between 2015 and 2018. The burning periods were divided into early (carried out during the rainy season), modal or “fruiting” (carried out during the transition between seasons) and late (carried out in the middle of the dry season, when the grasses are dormant). The results corroborate the traditional knowledge, demonstrating that the modal burns result in higher rates of reproduction and fruit production. Late burns showed intermediate results, followed by early burns. We conclude that management strategies based mainly on forage production, which are usually applied in savannas populated by grazing ungulates, may not be the best management strategy for South American savannas. The effects of fire on fruit plants, which have a particular phenologicalsynchronization with the fauna cycle, also need to be observed during the prescription of burns.

Keywords: cerrado biome, fire regimes, native fruits, prescribed burns

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Authors: Dibyendu Rakshit, Santosh K. Sarkar

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The study illustrates a comprehensive account of large-scale changes plankton community structure in relevance to water quality characteristics due to anthropogenic stresses, mainly concerned for Annual Gangasagar Festival (AGF) at the southern tip of Sagar Island of Indian Sundarban wetland for 3-year duration (2012-2014; n=36). This prograding, vulnerable and tide-dominated megadelta has been formed in the estuarine phase of the Hooghly Estuary infested by largest continuous tract of luxurious mangrove forest, enriched with high native flora and fauna. The sampling strategy was designed to characterize the changes in plankton community and water quality considering three diverse phases, namely during festival period (January) and its pre - (December) as well as post (February) events. Surface water samples were collected for estimation of different environmental variables as well as for phytoplankton and microzooplankton biodiversity measurement. The preservation and identification techniques of both biotic and abiotic parameters were carried out by standard chemical and biological methods. The intensive human activities lead to sharp ecological changes in the context of poor water quality index (WQI) due to high turbidity (14.02±2.34 NTU) coupled with low chlorophyll a (1.02±0.21 mg m-3) and dissolved oxygen (3.94±1.1 mg l-1), comparing to pre- and post-festival periods. Sharp reduction in abundance (4140 to 2997 cells l-1) and diversity (H′=2.72 to 1.33) of phytoplankton and microzooplankton tintinnids (450 to 328 ind l-1; H′=4.31 to 2.21) was very much pronounced. The small size tintinnid (average lorica length=29.4 µm; average LOD=10.5 µm) composed of Tintinnopsis minuta, T. lobiancoi, T. nucula, T. gracilis are predominant and reached some of the greatest abundances during the festival period. Results of ANOVA revealed a significant variation in different festival periods with phytoplankton (F= 1.77; p=0.006) and tintinnid abundance (F= 2.41; P=0.022). RELATE analyses revealed a significant correlation between the variations of planktonic communities with the environmental data (R= 0.107; p= 0.005). Three distinct groups were delineated from principal component analysis, in which a set of hydrological parameters acted as the causative factor(s) for maintaining diversity and distribution of the planktonic organisms. The pronounced adverse impact of anthropogenic stresses on plankton community could lead to environmental deterioration, disrupting the productivity of benthic and pelagic ecosystems as well as fishery potentialities which directly related to livelihood services. The festival can be considered as multiple drivers of changes in relevance to beach erosion, shoreline changes, pollution from discarded plastic and electronic wastes and destruction of natural habitats resulting loss of biodiversity. In addition, deterioration in water quality was also evident from immersion of idols, causing detrimental effects on aquatic biota. The authors strongly recommend for adopting integrated scientific and administrative strategies for resilience, sustainability and conservation of this megadelta.

Keywords: Gangasagar festival, phytoplankton, Sundarban megadelta, tintinnid

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Authors: Serap Durkut, A. Eser Elcin, Y. Murat Elcin

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Stimuli-sensitive polymeric hydrogels have received extensive attention in the biomedical field due to their sensitivity to physical and chemical stimuli (temperature, pH, ionic strength, light, etc.). This study describes the rheological properties of a novel thermoresponsive poly(N-vinylcaprolactam)-g-collagen hydrogel. In the study, we first synthesized a facile and novel synthetic carboxyl group-terminated thermo-responsive poly(N-vinylcaprolactam)-COOH (PNVCL-COOH) via free radical polymerization. Further, this compound was effectively grafted with native collagen, by utilizing the covalent bond between the carboxylic acid groups at the end of the chains and amine groups of the collagen using cross-linking agent (EDC/NHS), forming PNVCL-g-Col. Newly-formed hybrid hydrogel displayed novel properties, such as increased mechanical strength and thermoresponsive characteristics. PNVCL-g-Col showed low critical solution temperature (LCST) at 38ºC, which is very close to the body temperature. Rheological studies determine structural–mechanical properties of the materials and serve as a valuable tool for characterizing. The rheological properties of hydrogels are described in terms of two dynamic mechanical properties: the elastic modulus G′ (also known as dynamic rigidity) representing the reversible stored energy of the system, and the viscous modulus G″, representing the irreversible energy loss. In order to characterize the PNVCL-g-Col, the rheological properties were measured in terms of the function of temperature and time during phase transition. Below the LCST, favorable interactions allowed the dissolution of the polymer in water via hydrogen bonding. At temperatures above the LCST, PNVCL molecules within PNVCL-g-Col aggregated due to dehydration, causing the hydrogel structure to become dense. When the temperature reached ~36ºC, both the G′ and G″ values crossed over. This indicates that PNVCL-g-Col underwent a sol-gel transition, forming an elastic network. Following temperature plateau at 38ºC, near human body temperature the sample displayed stable elastic network characteristics. The G′ and G″ values of the PNVCL-g-Col solutions sharply increased at 6-9 minute interval, due to rapid transformation into gel-like state and formation of elastic networks. Copolymerization with collagen leads to an increase in G′, as collagen structure contains a flexible polymer chain, which bestows its elastic properties. Elasticity of the proposed structure correlates with the number of intermolecular cross-links in the hydrogel network, increasing viscosity. However, at 8 minutes, G′ and G″ values sharply decreased for pure collagen solutions due to the decomposition of the elastic and viscose network. Complex viscosity is related to the mechanical performance and resistance opposing deformation of the hydrogel. Complex viscosity of PNVCL-g-Col hydrogel was drastically changed with temperature and the mechanical performance of PNVCL-g-Col hydrogel network increased, exhibiting lesser deformation. Rheological assessment of the novel thermo-responsive PNVCL-g-Col hydrogel, exhibited that the network has stronger mechanical properties due to both permanent stable covalent bonds and physical interactions, such as hydrogen- and hydrophobic bonds depending on temperature.

Keywords: poly(N-vinylcaprolactam)-g-collagen, thermoresponsive polymer, rheology, elastic modulus, stimuli-sensitive

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Authors: Inna Kornienko, Svetlana Guryeva, Anatoly Shekhter, Elena Petersen

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Transplantation is an effective treatment option for patients suffering from different end-stage diseases. However, it is plagued by a constant shortage of donor organs and the subsequent need of a lifelong immunosuppressive therapy for the patient. Currently some researchers look towards using of pig organs to replace human organs for transplantation since the matrix derived from porcine organs is a convenient substitute for the human matrix. As an initial step to create a new ex vivo tissue engineered model, optimized protocols have been created to obtain organ-specific acellular matrices and evaluated their potential as tissue engineered scaffolds for culture of normal cells and tumor cell lines. These protocols include decellularization by perfusion in a bioreactor system and immersion-agitation on an orbital shaker with use of various detergents (SDS, Triton X-100) and freezing. Complete decellularization – in terms of residual DNA amount – is an important predictor of probability of immune rejection of materials of natural origin. However, the signs of cellular material may still remain within the matrix even after harsh decellularization protocols. In this regard, the matrices obtained from tissues of low-immunogenic pigs with α3Galactosyl-tranferase gene knock out (GalT-KO) may be a promising alternative to native animal sources. The research included a study of induced effect of frozen and fresh fragments of GalT-KO skin on healing of full-thickness plane wounds in 80 rats. Commercially available wound dressings (Ksenoderm, Hyamatrix and Alloderm) as well as allogenic skin were used as a positive control and untreated wounds were analyzed as a negative control. The results were evaluated on the 4th day after grafting, which corresponds to the time of start of normal wound epithelization. It has been shown that a non-specific immune response in models treated with GalT-Ko pig skin was milder than in all the control groups. Research has been performed to measure technical skin characteristics: stiffness and elasticity properties, corneometry, tevametry, and cutometry. These metrics enabled the evaluation of hydratation level, corneous layer husking level, as well as skin elasticity and micro- and macro-landscape. These preliminary data may contribute to development of personalized transplantable organs from GalT-Ko pigs with significantly limited potential of immune rejection. By applying growth factors to a decellularized skin sample it is possible to achieve various regenerative effects based on the particular situation. In this particular research BMP2 and Heparin-binding EGF-like growth factor have been used. Ideally, a bioengineered organ must be biocompatible, non-immunogenic and support cell growth. Porcine organs are attractive for xenotransplantation if severe immunologic concerns can be bypassed. The results indicate that genetically modified pig tissues with knock-outed α3Galactosyl-tranferase gene may be used for production of low-immunogenic matrix suitable for transplantation.

Keywords: decellularization, low-immunogenic, matrix, scaffolds, transplants

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Authors: Rodriguez Diego, Del Valle Martin, Hargreaves Matias, Riveros Jose Luis

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Nutrient deficiencies due to a diet low in quantitative and qualitative terms, are prevalent throughout the developing world, especially in sub-Saharan Africa. Children and women of childbearing age are especially vulnerable. Limited availability, access and intake of animal foods at home and lack of knowledge about their value in the diet and the role they play in health, contribute to poor diet quality. Poor bioavailability of micronutrients in diets based on foods high in fiber and phytates, the low content of some micronutrients in these foods are further factors to consider. Goats are deeply embedded in almost every Sub-Saharan African rural culture, generally kept for their milk, meat, hair or leather. Goats have played an important role in African social life, especially in food security. Goat meat has good properties for human wellbeing, with a special role in lower income households. It has a high-quality protein (20 protein g/100 meat g) including all essential amino acids, good unsaturated/satured fatty acids relationship, and it is an important B-vitamin source with high micronutrients bioavailability. Mozambique has major food security problems, with poor food access and utilization, undiversified diets, chronic poverty and child malnutrition. Our objective was to design a nutritional intervention based on a dietary diversification, nutritional education, cultural beliefs and local resources, aimed to strengthen food security of children at Barrio Broma village (15°43'58.78"S; 32°46'7.27"E) in Chitima, Mozambique. Two surveys were conducted first of socio-productive local databases and then to 100 rural households about livelihoods, food diversity and anthropometric measurements in children under 5 years. Our results indicate that the main economic activity is goat production, based on a native breed with two deliveries per year in the absence of any management. Adult goats weighted 27.2±10.5 kg and raised a height of 63.5±3.8 cm. Data showed high levels of poverty, with a food diversity score of 2.3 (0-12 points), where only 30% of households consume protein and 13% iron, zinc, and B12 vitamin. The main constraints to food security were poor access to water and low income to buy food. Our dietary intervention was based on improving diet quality by increasing the access to dried goat meat, fresh vegetables, and legumes, and its utilization by a nutritional education program. This proposal was based on local culture and living conditions characterized by the absence of electricity power and drinkable water. The drying process proposed would secure the food maintenance under local conditions guaranteeing food safety for a longer period. Additionally, an ancient local drying technique was rescued and used. Moreover, this kind of dietary intervention would be the most efficient way to improve the infant nutrition by delivering macro and micronutrients on time to these vulnerable populations.

Keywords: child malnutrition, dietary diversification, food security, goat meat

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Authors: Muna Shafiq

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Growth in international immigration is a reflection of increased migration patterns in Canada and in other parts of the world. Canada continues to promote itself as a bilingual country, yet the bilingual French and English population numbers do not reflect this platform. Each province’s integration policies focus only on second language learning of either English or French. Moreover, since English Canadians outnumber French Canadians, maintaining, much less increasing, English-French bilingualism appears unrealistic. One solution to increasing Canadian bilingualism requires creating intercultural communication initiatives between youth in Quebec and the rest of Canada. Specifically, the focus is on active, experiential learning, where intercultural competencies develop outside traditional classroom settings. The target groups are Generation Y Millennials and Generation Z Linksters, the next generations in the career and parenthood lines. Today, Canada’s education system, like many others, must continually renegotiate lines between programs it offers its immigrant and native communities. While some purists or right-wing nationalists would disagree, the survival of bilingualism in Canada has little to do with reducing immigration. Children and youth immigrants play a valuable role in increasing Canada’s French and English speaking communities. For instance, a focus on more immersion, over core French education programs for immigrant children and youth would not only increase bilingual rates; it would develop meaningful intercultural attachments between Canadians. Moreover, a vigilant increase of funding in French immersion programs is critical, as are new initiatives that focus on experiential language learning for students in French and English language programs. A favorable argument supports the premise that other than French-speaking students in Québec and elsewhere in Canada, second and third generation immigrant students are excellent ambassadors to promote bilingualism in Canada. Most already speak another language at home and understand the value of speaking more than one language in their adopted communities. Their dialogue and participation in experiential language exchange workshops are necessary. If the proposed exchanges take place inter-provincially, the momentum to increase collective regional voices increases. This regional collectivity can unite Canadians differently than nation-targeted initiatives. The results from an experiential youth exchange organized in 2017 between students at the crossroads of Generation Y and Generation Z in Vancouver and Quebec City respectively offer a promising starting point in assessing the strength of bringing together different regional voices to promote bilingualism. Code-switching between standard, international French Vancouver students, learn in the classroom versus more regional forms of Quebec French spoken locally created regional connectivity between students. The exchange was equally rewarding for both groups. Increasing their appreciation for each other’s regional differences allowed them to contribute actively to their social and emotional development. Within a sociolinguistic frame, this proposed model of experiential learning does not focus on hands-on work experience. However, the benefits of such exchanges are as valuable as work experience initiatives developed in experiential education. Students who actively code switch between French and English in real, not simulated contexts appreciate bilingualism more meaningfully and experience its value in concrete terms.

Keywords: experiential learning, intercultural communication, social and emotional learning, sociolinguistic code-switching

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Authors: Omaymah Al-Otoom, Rajesh Mehta

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Introduction: Migration is an established risk factor in the development of schizophrenia and other forms of psychosis. The exposure to different social adversities, including social isolation, discrimination, and economic stress, is thought to contribute to elevated rates of psychosis in immigrants and their children. We present a case of resistant schizophrenia in an immigrant adolescent. Case: The patient is a 15-year-old male immigrant. In October 2021, the patient was admitted for irritability, suicidal ideations, and hallucinations. He was treated with Fluoxetine 10 mg daily for irritability. In November 2021, he presented with similar manifestations. Fluoxetine was discontinued, and Risperidone 1 mg at bedtime was started for psychotic symptoms. In March 2022, he presented with commanding auditory hallucinations (voices telling him that people were going to kill his father). Risperidone was gradually increased to 2.5 mg twice daily for hallucinations. The outpatient provider discontinued Risperidone and started Olanzapine 7.5 mg and Lurasidone 40 mg daily. In August 2022, he presented with worsening paranoia due to medication non-adherence. The patient had limited improvement on medications. In October 2022, the patient presented to the ED for visual hallucinations and aggression towards the family. His medications were Olanzapine 10 mg daily, Lurasidone 60 mg daily, and Haloperidol 2.5 mg twice daily. In the ED, he received multiple as-needed medications and was placed in seclusion for his aggressive behavior. The patient showed a positive response to a higher dose of Olanzapine and decreased dose of Lurasidone. The patient was discharged home in stable condition. Two days after discharge, he was brought for bizarre behavior, visual hallucinations, and homicidal ideations at school. Due to concerns for potential antipsychotic side effects and poor response, Lurasidone and Olanzapine were discontinued, and he was discharged home on Haloperidol 5 mg in the morning and 15 mg in the evening. Clozapine treatment was recommended on an outpatient basis. He has no family history of psychotic disorders. He has no history of substance use. A medical workup was done, the electroencephalogram was normal, and the urine toxicology was negative. Discussion: Our patient was on three antipsychotics at some point with no improvement in his psychotic symptoms, which qualifies as treatment-resistant schizophrenia (TRP). It is well recognized that migrants are at higher risk of different psychiatric disorders, including posttraumatic stress disorder, affective disorders, schizophrenia, and psychosis. This is thought to be related to higher exposure to traumatic life events compared to the general population. In addition, migrants are more likely to experience poverty, separation from family members, and discrimination which could contribute to mental health issues. In one study, they found that people who migrated before the age of 18 had twice the risk of psychotic disorders compared to the native-born population. It is unclear whether migration increases the risk of treatment resistance. In a Canadian study, neither ethnicity nor migrant status was associated with treatment resistance; however, this study was limited by its small sample size. There is a need to implement psychiatric prevention strategies and outreach programs through research to mitigate the risk of mental health disorders among immigrants.

Keywords: psychosis, immigrant, adolescent, treatment resistant schizophrenia

Procedia PDF Downloads 88

Authors: Hamza Saeed, Khurram Iqbal Ahmad Khan

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Effective information management is crucial for any construction project and its success. Primary areas of information generation are either the construction site or the design office. There are different types of information required at different stages of construction involving various stakeholders creating complexity. There is a need for effective management of information flows to reduce uncertainty creating complexity. Nature provides a unique perspective in terms of dealing with complexity, in particular, information complexity. System dynamics methodology provides tools and techniques to address complexity. It involves modeling and simulation techniques that help address complexity. Nature has been dealing with complex systems since its creation 4.5 billion years ago. It has perfected its system by evolution, resilience towards sudden changes, and extinction of unadaptable and outdated species that are no longer fit for the environment. Nature has been accommodating the changing factors and handling complexity forever. Humans have started to look at their natural counterparts for inspiration and solutions for their problems. This brings forth the possibility of using a biomimetics approach to improve the management practices used in the construction sector. Ants inhabit different habitats. Cataglyphis and Pogonomyrmex live in deserts, Leafcutter ants reside in rainforests, and Pharaoh ants are native to urban developments of tropical areas. Detailed studies have been done on fifty species out of fourteen thousand discovered. They provide the opportunity to study the interactions in diverse environments to generate collective behavior. Animals evolve to better adapt to their environment. The collective behavior of ants emerges from feedback through interactions among individuals, based on a combination of three basic factors: The patchiness of resources in time and space, operating cost, environmental stability, and the threat of rupture. If resources appear in patches through time and space, the response is accelerating and non-linear, and if resources are scattered, the response follows a linear pattern. If the acquisition of energy through food is faster than energy spent to get it, the default is to continue with an activity unless it is halted for some reason. If the energy spent is rather higher than getting it, the default changes to stay put unless activated. Finally, if the environment is stable and the threat of rupture is low, the activation and amplification rate is slow but steady. Otherwise, it is fast and sporadic. To further study the effects and to eliminate the environmental bias, the behavior of four different ant species were studied, namely Red Harvester ants (Pogonomyrmex Barbatus), Argentine ants (Linepithema Humile), Turtle ants (Cephalotes Goniodontus), Leafcutter ants (Genus: Atta). This study aims to improve the information system in the construction sector by providing a guideline inspired by nature with a systems-thinking approach, using system dynamics as a tool. Identified factors and their interdependencies were analyzed in the form of a causal loop diagram (CLD), and construction industry professionals were interviewed based on the developed CLD, which was validated with significance response. These factors and interdependencies in the natural system corresponds with the man-made systems, providing a guideline for effective use and flow of information.

Keywords: biomimetics, complex systems, construction management, information management, system dynamics

Procedia PDF Downloads 113

Authors: Jyh-Ping Chen, Yu-Tin Lai

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Wound healing is a complicated process involving overlapping hemostasis, inflammation, proliferation, and maturation phases. Ideal wound dressings can replace native skin functions in full thickness skin wounds through faster healing rate and also by reducing scar formation. Poly(lactic-co-glycolic acid) (PLGA) is an U.S. FDA approved biodegradable polymer to be used as ideal wound dressing material. Several in vitro and in vivo studies have demonstrated the effectiveness of curcumin in decreasing the release of inflammatory cytokines, inhibiting enzymes associated with inflammations, and scavenging free radicals that are the major cause of inflammation during wound healing. Heparin has binding affinities to various growth factors. With the unique and beneficial features offered by those molecules toward the complex process of wound healing, we postulate a composite wound dressing constructed from PLGA, curcumin and heparin would be a good candidate to accelerate scarless wound healing. In this work, we use electrospinning to prepare curcumin-loaded aligned PLGA nanofibrous membranes (PC NFMs). PC NFMs were further subject to oxygen plasma modification and surfaced-grafted with heparin through carbodiimide-mediated covalent bond formation to prepare curcumin-loaded PLGA-g-heparin (PCH) NFMs. The nanofibrous membranes could act as three-dimensional scaffolds to attract fibroblast migration, reduce inflammation, and increase wound-healing related growth factors concentrations at wound sites. From scanning electron microscopy analysis, the nanofibers in each NFM are with diameters ranging from 456 to 479 nm and with alignment angles within  0.5°. The NFMs show high tensile strength and good water absorptivity and provide suitable pore size for nutrients/wastes transport. Exposure of human dermal fibroblasts to the extraction medium of PC or PCH NFM showed significant protective effects against hydrogen peroxide than PLGA NFM. In vitro wound healing assays also showed that the extraction medium of PCH NFM showed significantly better migration ability toward fibroblasts than PC NFM, which is further better than PLGA NFM. The in vivo healing efficiency of the NFMs was further evaluated by a full thickness excisional wound healing diabetic rat model. After 14 days, PCH NFMs exhibits 86% wound closure rate, which is significantly different from other groups (79% for PC and 73% for PLGA NFM). Real-time PCR analysis indicated PC and PCH NFMs down regulated anti-oxidative enzymes like glutathione peroxidase (GPx) and superoxide dismutase (SOD), which are well-known transcription factors involved in cellular inflammatory responses to stimuli. From histology, the wound area treated with PCH NFMs showed more vascular lumen formation from immunohistochemistry of α-smooth muscle actin. The wound site also had more collagen type III (65.8%) expression and less collagen type I (3.5%) expression, indicating scar-less wound healing. From Western blot analysis, the PCH NFM showed good affinity toward growth factors from increased concentration of transforming growth factor-β (TGF-β) and fibroblast growth factor-2 (FGF-2) at the wound site to accelerate wound healing. From the results, we suggest PCH NFM as a promising candidate for wound dressing applications.

Keywords: Curcumin, heparin, nanofibrous membrane, poly(lactic-co-glycolic acid) (PLGA), wound dressing

Procedia PDF Downloads 130

Authors: Tamal Roy, Anuradha Bhat

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Cognitive abilities aid fishes in foraging, avoiding predators & locating mates. Factors like predation pressure & habitat complexity govern learning & memory in fishes. This study aims to compare spatial learning & memory across four natural populations of zebrafish. Zebrafish, a small cyprinid inhabits a diverse range of freshwater habitats & this makes it amenable to studies investigating role of native environment in spatial cognitive abilities. Four populations were collected across India from waterbodies with contrasting ecological conditions. Habitat complexity of the water-bodies was evaluated as a combination of channel substrate diversity and diversity of vegetation. Experiments were conducted on populations under controlled laboratory conditions. A square shaped spatial testing arena (maze) was constructed for testing the performance of adult zebrafish. The square tank consisted of an inner square shaped layer with the edges connected to the diagonal ends of the tank-walls by connections thereby forming four separate chambers. Each of the four chambers had a main door in the centre. Each chamber had three sections separated by two windows. A removable coloured window-pane (red, yellow, green or blue) identified each main door. A food reward associated with an artificial plant was always placed inside the left-hand section of the red-door chamber. The position of food-reward and plant within the red-door chamber was fixed. A test fish would have to explore the maze by taking turns and locate the food inside the right-side section of the red-door chamber. Fishes were sorted from each population stock and kept individually in separate containers for identification. At a time, a test fish was released into the arena and allowed 20 minutes to explore in order to find the food-reward. In this way, individual fishes were trained through the maze to locate the food reward for eight consecutive days. The position of red door, with the plant and the reward, was shuffled every day. Following training, an intermission of four days was given during which the fishes were not subjected to trials. Post-intermission, the fishes were re-tested on the 13th day following the same protocol for their ability to remember the learnt task. Exploratory tendencies and latency of individuals to explore on 1st day of training, performance time across trials, and number of mistakes made each day were recorded. Additionally, mechanism used by individuals to solve the maze each day was analyzed across populations. Fishes could be expected to use algorithm (sequence of turns) or associative cues in locating the food reward. Individuals of populations did not differ significantly in latencies and tendencies to explore. No relationship was found between exploration and learning across populations. High habitat-complexity populations had higher rates of learning & stronger memory while low habitat-complexity populations had lower rates of learning and much reduced abilities to remember. High habitat-complexity populations used associative cues more than algorithm for learning and remembering while low habitat-complexity populations used both equally. The study, therefore, helped understand the role of natural ecology in explaining variations in spatial learning abilities across populations.

Keywords: algorithm, associative cue, habitat complexity, population, spatial learning

Procedia PDF Downloads 266

Authors: L. R. Valencia Pérez, J. M. Peña Aguilar, A. Lamadrid Álvarez, A. Pastrana Palma, H. F. Valencia Pérez, M. Vivanco Vargas

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This work proposes a Cooperation-Competitive (Coopetitive) approach that allows coordinated work among the Secretary of Public Education (SEP), the Autonomous University of Querétaro (UAQ) and government funds from National Council for Science and Technology (CONACYT) or some other international organizations. To work on an overall knowledge transfer strategy with e-learning over the Cloud, where experts in junior high and high school education, working in multidisciplinary teams, perform analysis, evaluation, design, production, validation and knowledge transfer at large scale using a Cloud Computing platform. Allowing teachers and students to have all the information required to ensure a homologated nationally knowledge of topics such as mathematics, statistics, chemistry, history, ethics, civism, etc. This work will start with a pilot test in Spanish and initially in two regional dialects Otomí and Náhuatl. Otomí has more than 285,000 speaking indigenes in Queretaro and Mexico´s central region. Náhuatl is number one indigenous dialect spoken in Mexico with more than 1,550,000 indigenes. The phase one of the project takes into account negotiations with indigenous tribes from different regions, and the Information and Communication technologies to deliver the knowledge to the indigenous schools in their native dialect. The methodology includes the following main milestones: Identification of the indigenous areas where Otomí and Náhuatl are the spoken dialects, research with the SEP the location of actual indigenous schools, analysis and inventory or current schools conditions, negotiation with tribe chiefs, analysis of the technological communication requirements to reach the indigenous communities, identification and inventory of local teachers technology knowledge, selection of a pilot topic, analysis of actual student competence with traditional education system, identification of local translators, design of the e-learning platform, design of the multimedia resources and storage strategy for “Cloud Computing”, translation of the topic to both dialects, Indigenous teachers training, pilot test, course release, project follow up, analysis of student requirements for the new technological platform, definition of a new and improved proposal with greater reach in topics and regions. Importance of phase one of the project is multiple, it includes the proposal of a working technological scheme, focusing in the cultural impact in Mexico so that indigenous tribes can improve their knowledge about new forms of crop improvement, home storage technologies, proven home remedies for common diseases, ways of preparing foods containing major nutrients, disclose strengths and weaknesses of each region, communicating through cloud computing platforms offering regional products and opening communication spaces for inter-indigenous cultural exchange.

Keywords: Mexicans indigenous tribes, education, knowledge transfer, cloud computing, otomi, Náhuatl, language

Procedia PDF Downloads 371

Authors: Medan Isaeva, Anna Degtyareva

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Context: Biliary atresia is a common reason for liver transplants in children, and the Kasai procedure can potentially be successful in avoiding the need for transplantation. However, it is important to identify factors that influence surgical outcomes in order to optimize treatment and improve patient outcomes. Research aim: The aim of this study was to develop prognostic models to assess the outcomes of the Kasai procedure in children with biliary atresia. Methodology: This retrospective study analyzed data from 166 children with biliary atresia who underwent the Kasai procedure between 2002 and 2021. The effectiveness of the operation was assessed based on specific criteria, including post-operative stool color, jaundice reduction, and bilirubin levels. The study involved a comparative analysis of various parameters, such as gestational age, birth weight, age at operation, physical development, liver and spleen sizes, and laboratory values including bilirubin, ALT, AST, and others, measured pre- and post-operation. Ultrasonographic evaluations were also conducted pre-operation, assessing the hepatobiliary system and related quantitative parameters. The study was carried out by two experienced specialists in pediatric hepatology. Comparative analysis and multifactorial logistic regression were used as the primary statistical methods. Findings: The study identified several statistically significant predictors of a successful Kasai procedure, including the presence of the gallbladder and levels of cholesterol and direct bilirubin post-operation. A detectable gallbladder was associated with a higher probability of surgical success, while elevated post-operative cholesterol and direct bilirubin levels were indicative of a reduced chance of positive outcomes. Theoretical importance: The findings of this study contribute to the optimization of treatment strategies for children with biliary atresia undergoing the Kasai procedure. By identifying early predictive signs of success, clinicians can modify treatment plans and manage patient care more effectively and proactively. Data collection and analysis procedures: Data for this analysis were obtained from the health records of patients who received the Kasai procedure. Comparative analysis and multifactorial logistic regression were employed to analyze the data and identify significant predictors. Question addressed: The study addressed the question of identifying predictive factors for the success of the Kasai procedure in children with biliary atresia. Conclusion: The developed prognostic models serve as valuable tools for early detection of patients who are less likely to benefit from the Kasai procedure. This enables clinicians to modify treatment plans and manage patient care more effectively and proactively. Potential limitations of the study: The study has several limitations. Its retrospective nature may introduce biases and inconsistencies in data collection. Being single centered, the results might not be generalizable to wider populations due to variations in surgical and postoperative practices. Also, other potential influencing factors beyond the clinical, laboratory, and ultrasonographic parameters considered in this study were not explored, which could affect the outcomes of the Kasai operation. Future studies could benefit from including a broader range of factors.

Keywords: biliary atresia, kasai operation, prognostic model, native liver survival

Procedia PDF Downloads 25

Authors: Prem Kumar, Anuj Tyagi, Harsh Panwar, Vaneet Inder Kaur

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Though aquaculture started as an occupation for poor and weak farmers for livelihood, it has now acquired the shape of one of the biggest industry to grow live protein in the form of aquatic organisms. Industrialization of the aquaculture sector has led to intensification resulting in stress on aquatic organisms and frequent disease outbreaks leading to huge economic impacts. Indiscriminate use of antibiotics as growth promoter and prophylactic agent in aquaculture has resulted in rapid emergence and spread of antibiotic resistance in bacterial pathogens. Over the past few years, use of probiotics (as an alternative of antibiotics) in aquaculture has gained attention due to their immunostimulant and growth promoting properties. It has now well known that after administration, a probiotic bacterium has to compete and establish itself against native microbiota to show its eventual beneficial properties. Due to their non-fish origin, commercial probiotics sometimes may display poor probiotic functionalities and antagonistic effects. Thus, isolation and characterization of probiotic bacteria from same fish host is very much necessary. In this study, attempts were made to isolate potent probiotic lactic acid bacteria (LAB) from intestinal microflora of rohu fish. Twenty-five experimental rohu fishes (mean weight 400 ± 20gm, mean standard length 20 ± 3cm) were used in the study to collect fish gut after dissection in a sterile condition. A total of 150 tentative LAB isolates from selective agar media (de Man-Rogosa-Sharpe (MRS)) were screened for their antimicrobial activity against Aeromonas hydrophila and Microccocus leuteus. A total of 17 isolates, identified as Lactobacillus plantarum and Lactococcus lactis, identified by biochemical tests and PCR amplification and sequencing of 16S rRNA gene fragment, displayed promising antimicrobial activity against both the pathogens. Two isolates from each species (FLB1, FLB2 from L. plantarum; and FLC1, FLC2 from L. lactis) were subjected to downstream probiotic potential characterization. These isolates were compared in vitro for their hemolytic activity, acid and bile tolerance for growth kinetics, auto-aggregation, cell-surface hydrophobicity against xylene, and chloroform, tolerance to phenol, cell adhesion, and safety parameters (by intraperitoneal and intramuscular injections). None of the tested isolates showed any hemolytic activity indicating their potential safety. Moreover, these isolates were tolerant to 0.3% bile (75-82% survival), phenol stress (96-99% survival) with 100% viability at pH 3 over a period of 3 h. Antibiotic sensitivity test revealed that all the tested LAB isolates were resistant to vancomycin, gentamicin, streptomycin, and erythromycin and sensitive to Erythromycin, Chloramphenicol, Ampicillin, Trimethoprim, and Nitrofurantoin. Tetracycline resistance was found in L. plantarum (FLB1 and FLB2 isolates), whereas L. lactis were susceptible to it. Intramuscular and intraperitoneal challenges to fingerlings of rohu fish (5 ± 1gm weight) with FLB1 showed no pathogenicity and occurrence of disease symptoms in fishes over an observation period of 7 days. The results revealed FLB1 as a potential probiotic candidate for aquaculture application among other isolates.

Keywords: aquaculture, Lactobacillus plantarum, Lactococcus lactis, probiotics

Procedia PDF Downloads 109

Authors: Adeline Meriaux, Claire Gaiani, Jennifer Burgain, Frantz Fournier, Lionel Muniglia, Jérémy Petit

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Spray-drying process is widely used for the production of dairy powders for food and pharmaceuticals industries. It involves the atomization of a liquid feed into fine droplets, which are subsequently dried through contact with a hot air flow. The resulting powders permit transportation cost reduction and shelf life increase but can also exhibit various interesting functionalities (flowability, solubility, protein modification or acid gelation), depending on operating conditions and milk composition. Indeed, particles porosity, surface composition, lactose crystallization, protein denaturation, protein association or crust formation may change. Links between spray-drying conditions and physicochemical and functional properties of powders were investigated by a design of experiment methodology and analyzed by principal component analysis. Quadratic models were developed, and multicriteria optimization was carried out by the use of genetic algorithm. At the time of abstract submission, verification spray-drying trials are ongoing. To perform experiments, milk from dairy farm was collected, skimmed, froze and spray-dried at different air pressure (between 1 and 3 bars) and outlet temperature (between 75 and 95 °C). Dry matter, minerals content and proteins content were determined by standard method. Solubility index, absorption index and hygroscopicity were determined by method found in literature. Particle size distribution were obtained by laser diffraction granulometry. Location of the powder color in the Cielab color space and water activity were characterized by a colorimeter and an aw-value meter, respectively. Flow properties were characterized with FT4 powder rheometer; in particular compressibility and shearing test were performed. Air pressure and outlet temperature are key factors that directly impact the drying kinetics and powder characteristics during spray-drying process. It was shown that the air pressure affects the particle size distribution by impacting the size of droplet exiting the nozzle. Moreover, small particles lead to more cohesive powder and less saturated color of powders. Higher outlet temperature results in lower moisture level particles which are less sticky and can explain a spray-drying yield increase and the higher cohesiveness; it also leads to particle with low water activity because of the intense evaporation rate. However, it induces a high hygroscopicity, thus, powders tend to get wet rapidly if they are not well stored. On the other hand, high temperature provokes a decrease of native serum proteins which is positively correlated to gelation properties (gel point and firmness). Partial denaturation of serum proteins can improve functional properties of powder. The control of air pressure and outlet temperature during the spray-drying process significantly affects the physicochemical and functional properties of powder. This study permitted to better understand the links between physicochemical and functional properties of powder, to identify correlations between air pressure and outlet temperature. Therefore, mathematical models have been developed and the use of genetic algorithm will allow the optimization of powder functionalities.

Keywords: dairy powders, spray-drying, powders functionalities, design of experiment

Procedia PDF Downloads 47

Authors: Adeline Meriaux, Claire Gaiani, Jennifer Burgain, Frantz Fournier, Lionel Muniglia, Jérémy Petit

Abstract:

Spray-drying process is widely used for the production of dairy powders for food and pharmaceuticals industries. It involves the atomization of a liquid feed into fine droplets, which are subsequently dried through contact with a hot air flow. The resulting powders permit transportation cost reduction and shelf life increase but can also exhibit various interesting functionalities (flowability, solubility, protein modification or acid gelation), depending on operating conditions and milk composition. Indeed, particles porosity, surface composition, lactose crystallization, protein denaturation, protein association or crust formation may change. Links between spray-drying conditions and physicochemical and functional properties of powders were investigated by a design of experiment methodology and analyzed by principal component analysis. Quadratic models were developed, and multicriteria optimization was carried out by the use of genetic algorithm. At the time of abstract submission, verification spray-drying trials are ongoing. To perform experiments, milk from dairy farm was collected, skimmed, froze and spray-dried at different air pressure (between 1 and 3 bars) and outlet temperature (between 75 and 95 °C). Dry matter, minerals content and proteins content were determined by standard method. Solubility index, absorption index and hygroscopicity were determined by method found in literature. Particle size distribution were obtained by laser diffraction granulometry. Location of the powder color in the Cielab color space and water activity were characterized by a colorimeter and an aw-value meter, respectively. Flow properties were characterized with FT4 powder rheometer; in particular, compressibility and shearing test were performed. Air pressure and outlet temperature are key factors that directly impact the drying kinetics and powder characteristics during spray-drying process. It was shown that the air pressure affects the particle size distribution by impacting the size of droplet exiting the nozzle. Moreover, small particles lead to more cohesive powder and less saturated color of powders. Higher outlet temperature results in lower moisture level particles which are less sticky and can explain a spray-drying yield increase and the higher cohesiveness; it also leads to particle with low water activity because of the intense evaporation rate. However, it induces a high hygroscopicity, thus, powders tend to get wet rapidly if they are not well stored. On the other hand, high temperature provokes a decrease of native serum proteins, which is positively correlated to gelation properties (gel point and firmness). Partial denaturation of serum proteins can improve functional properties of powder. The control of air pressure and outlet temperature during the spray-drying process significantly affects the physicochemical and functional properties of powder. This study permitted to better understand the links between physicochemical and functional properties of powder to identify correlations between air pressure and outlet temperature. Therefore, mathematical models have been developed, and the use of genetic algorithm will allow the optimization of powder functionalities.

Keywords: dairy powders, spray-drying, powders functionalities, design of experiment

Procedia PDF Downloads 41

Authors: Saraswati Kerketta

Abstract:

Tea industry is one of the oldest organised sector of India. It employs roughly 1.5 million people directly. Since the last decade Indian tea industry, especially in the northern region is experiencing worst crisis in the post-independence period. Due to many reason the prices of tea show steady decline. The workers are paid one of the lowest wage in tea industry in the world (1.5$ a day) below the UN's $2 a day for extreme poverty. The workers rely on addition benefits from plantation which includes food, housing and medical facilities. These have been effective means of enslavement of generations of labourers by the owners. There is hardly any change in the tea estates where the owners determine the fate of workers. When the tea garden is abandoned or is closed all the facilities disappear immediately. The workers are the descendants of tribes from central India also known as 'tea tribes'. Alienated from their native place, the geographical and social isolation compounded their vulnerability of these people. The economy of the region being totally dependent on tea has resulted in absolute unemployment for the workers of these tea gardens. With no other livelihood and no land to grow food, thousands of workers faced hunger and starvation. The Plantation Labour Act which ensures the decent working and living condition is violated continuously. The labours are forced to migrate and are also exposed to the risk of human trafficking. Those who are left behind suffers from starvation, malnutrition and disease. The condition in the sick tea plantation is no better. Wage are not paid regularly, subsidised food, fuel are also not supplied properly. Health care facilities are in very bad shape. Objectives: • To study the socio-cultural and demographic characteristics of the tea garden labourers in the study area. • To examine the social situation of workers in sick estates in dooars region. • To assess the magnitude of deprivation the impact of economic crisis on abandoned and closed tea estates in the region. Data Base: The study is based on data collected from field survey. Methods: Quantative: Cross-Tabulation, Regression analysis. Qualitative: Household Survey, Focussed Group Discussion, In-depth interview of key informants. Findings: Purchasing power parity has declined since in last three decades. There has been many fold increase in migration. Males migrates long distance towards central and west and south India. Females and children migrates both long and short distance. No one has reported to migrate back to the place of origin of their ancestors. Migrant males work mostly as construction labourers and as factory workers whereas females and children work as domestic help and construction labourers. In about 37 cases either they haven't contacted their families in last six months or are not traceable. The families with single earning members are more likely to migrate. Burden of disease and the duration of sickness, abandonment and closure of plantation are closely related. Death tolls are likely to rise 1.5 times in sick tea gardens and three times in closed tea estates. Sixty percent of the people are malnourished in the sick tea gardens and more than eighty five per cent in abandoned and sick tea gardens.

Keywords: migration, trafficking, starvation death, tea garden workers

Procedia PDF Downloads 356

Authors: Elizabeth J. Currie, Fernando Ortega Perez

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—‘MEDICINE’ is a new project funded under the EC Horizon 2020 Marie-Sklodowska Curie Actions, to determine concepts of health and healing from a culturally specific indigenous context, using a framework of interdisciplinary methods which integrates archaeological-historical, ethnographic and modern health sciences approaches. The study will generate new theoretical and methodological approaches to model how peoples survive and adapt their traditional belief systems in a context of alien cultural impacts. In the immediate wake of the conquest of Peru by invading Spanish armies and ideology, native Andeans responded by forming the Taki Onkoy millenarian movement, which rejected European philosophical and ontological teachings, claiming “you make us sick”. The study explores how people’s experience of their world and their health beliefs within it, is fundamentally shaped by their inherent beliefs about the nature of being and identity in relation to the wider cosmos. Cultural and health belief systems and related rituals or behaviors sustain a people’s sense of identity, wellbeing and integrity. In the event of dislocation and persecution these may change into devolved forms, which eventually inter-relate with ‘modern’ biomedical systems of health in as yet unidentified ways. The development of new conceptual frameworks that model this process will greatly expand our understanding of how people survive and adapt in response to cultural trauma. It will also demonstrate the continuing role, relevance and use of TM in present-day indigenous communities. Studies will first be made of relevant pre-Colombian material culture, and then of early colonial period ethnohistorical texts which document the health beliefs and ritual practices still employed by indigenous Andean societies at the advent of the 17^th century Jesuit campaigns of persecution - ‘Extirpación de las Idolatrías’. Core beliefs drawn from these baseline studies will then be used to construct a questionnaire about current health beliefs and practices to be taken into the study population of indigenous Quechua peoples in the northern Andean region of Ecuador. Their current systems of knowledge and medicine have evolved within complex historical contexts of both the conquest by invading Inca armies in the late 15^th century, followed a generation later by Spain, into new forms. A new model will be developed of contemporary  Andean concepts of health, illness and healing demonstrating  the way these have changed through time. With this, a ‘policy tool’ will be constructed as a bridhging facility into contemporary global scenarios relevant to other Indigenous, First Nations, and migrant peoples to provide a means through which their traditional health beliefs and current needs may be more appropriately understood and met. This paper presents findings from the first analytical phases of the work based upon the study of the literature and the archaeological records. The study offers a novel perspective and methods in the development policies sensitive to indigenous and minority people’s health needs.

Keywords: Andean ethnomedicine, Andean health beliefs, health beliefs models, traditional medicine

Procedia PDF Downloads 322

Authors: Jesus Carretero, Mario Vasile, Javier Garcia-Blas, Felix Garcia-Carballeira

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Supporting cross-border secure and reliable exchange of data and documents and to promote data interoperability is critical for Europe to enhance sector (like eFinance, eJustice and eHealth). This work presents the status and results of the European Project MADE, a Research Project funded by Connecting Europe facility Programme, to provide secure e-invoicing and e-document exchange systems among Europe countries in compliance with the eIDAS Regulation (Regulation EU 910/2014 on electronic identification and trust services). The main goal of MADE is to develop six new AS4 Access Points and SMP in Europe to provide secure document exchanges using the eDelivery DSI (Digital Service Infrastructure) amongst both private and public entities. Moreover, the project demonstrates the feasibility and interest of the solution provided by providing several months of interoperability among the providers of the six partners in different EU countries. To achieve those goals, we have followed a methodology setting first a common background for requirements in the partner countries and the European regulations. Then, the partners have implemented access points in each country, including their service metadata publisher (SMP), to allow the access to their clients to the pan-European network. Finally, we have setup interoperability tests with the other access points of the consortium. The tests will include the use of each entity production-ready Information Systems that process the data to confirm all steps of the data exchange. For the access points, we have chosen AS4 instead of other existing alternatives because it supports multiple payloads, native web services, pulling facilities, lightweight client implementations, modern crypto algorithms, and more authentication types, like username-password and X.509 authentication and SAML authentication. The main contribution of MADE project is to open the path for European companies to use eDelivery services with cross-border exchange of electronic documents following PEPPOL (Pan-European Public Procurement Online) based on the e-SENS AS4 Profile. It also includes the development/integration of new components, integration of new and existing logging and traceability solutions and maintenance tool support for PKI. Moreover, we have found that most companies are still not ready to support those profiles. Thus further efforts will be needed to promote this technology into the companies. The consortium includes the following 9 partners. From them, 2 are research institutions: University Carlos III of Madrid (Coordinator), and Universidad Politecnica de Valencia. The other 7 (EDICOM, BIZbrains, Officient, Aksesspunkt Norge, eConnect, LMT group, Unimaze) are private entities specialized in secure delivery of electronic documents and information integration brokerage in their respective countries. To achieve cross-border operativity, they will include AS4 and SMP services in their platforms according to the EU Core Service Platform. Made project is instrumental to test the feasibility of cross-border documents eDelivery in Europe. If successful, not only einvoices, but many other types of documents will be securely exchanged through Europe. It will be the base to extend the network to the whole Europe. This project has been funded under the Connecting Europe Facility Agreement number: INEA/CEF/ICT/A2016/1278042. Action No: 2016-EU-IA-0063.

Keywords: security, e-delivery, e-invoicing, e-delivery, e-document exchange, trust

Procedia PDF Downloads 225

Authors: Lewis Mehl-Madrona, Barbara Mainguy, Patrick McFarlane

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Introduction: There are many risk factors for attempting suicide, including young age, “minority stress,” which would include Transgender and Gender Diverse orientations (TGD). The rate of TGD youths for suicide attempts is 3 times higher than heterosexual cis-gender youth. Half of TGD youth have seriously contemplated taking their own lives; of those, about half attempted suicide; and 18% of the TGD teenagers reported suicidal thoughts linked to their gender identity. Native American TGD have a six times higher suicide attempt rate. Conventional mental health has not generally helped these individuals. Stigma and discrimination contribute to healthcare disparities. Storytelling plays a crucial role in the development of human culture and individual identities. Sharing narrative artwork, creative writing, and personal stories allow people to build trust and to share their vulnerabilities. This helps people become aware of themselves in relation to others and gain a sense of comfort that their stories are similar; they may also be transformed in the process. Art provides a means to reach people who are otherwise difficult to engage in services. Methods: TGD individuals are recruited through a snowballing procedure. Following a life story interview, participants complete a scale of gender dysphoria, identification with conventional masculinity, patient-reported anxiety, and depression measure, and a quality-of-life scale. The interview completes the Columbia Suicide Scale. Following this, an artist and a therapist works with the participant to create a story related to their gender identity using the six-part story method. This story is then rendered to an artists’ book, which combines narrative with art (drawings, collage, computer images, etc.) and can take the form of a graphic novella, a zine, or a comic book. The pages can range from plain to ornate, as can the covers. Participants describe their process of making the books as the work unfolds and then participate in an exit interview at the completion of their book, remarking on what has changed for them and how the process affected them. Results: Preliminary results show high levels of suicidal thoughts among this population, as expected. Participants participate enthusiastically in the life story interview process and in the construction of a story related to gender identity. They enthusiastically participate in the studio process of putting their story into the form of a graphic novel, zine, or comic book. Participants reported feeling more comfortable with their TGD identity after the process and more able to resist negative judgments of family members and society. Suicidal thoughts diminish, and participants reported improved emotional wellbeing. Quantitative analysis of questionnaire data is underway Conclusions: A process in which narrative therapy is combined with art therapy shows promise for attracting and helping TGD individuals to reduce their risk for suicide without the stigma of going for mental health treatment. This process can be done outside of conventional mental health settings, on college and University campuses. This can provide an exciting alternative pathway for minority stressed and stigmatized individuals to engage in reflective, psychotherapeutic work without the trappings of psychotherapy or mental health treatment.

Keywords: minority stress, narrative process, artists' books, life story interview

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Authors: Yao-Ying Lai, Maria Pinango, Ashwini Deo

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While grammatical devices are essential to language processing, how comprehension utilizes cognitive mechanisms is less emphasized. This study addresses this issue by probing the complement coercion phenomenon: an entity-denoting complement following verbs like begin and finish receives an eventive interpretation. For example, (1) “The queen began the book” receives an agentive reading like (2) “The queen began [reading/writing/etc.…] the book.” Such sentences engender additional processing cost in real-time comprehension. The traditional account attributes this cost to an operation that coerces the entity-denoting complement to an event, assuming that these verbs require eventive complements. However, in closer examination, examples like “Chapter 1 began the book” undermine this assumption. An alternative, Structured Individual (SI) hypothesis, proposes that the complement following aspectual verbs (AspV; e.g. begin, finish) is conceptualized as a structured individual, construed as an axis along various dimensions (e.g. spatial, eventive, temporal, informational). The composition of an animate subject and an AspV such as (1) engenders an ambiguity between an agentive reading along the eventive dimension like (2), and a constitutive reading along the informational/spatial dimension like (3) “[The story of the queen] began the book,” in which the subject is interpreted as a subpart of the complement denotation. Comprehenders need to resolve the ambiguity by searching contextual information, resulting in additional cost. To evaluate the SI hypothesis, a questionnaire was employed. Method: Target AspV sentences such as “Shakespeare began the volume.” were preceded by one of the following types of context sentence: (A) Agentive-biasing, in which an event was mentioned (…writers often read…), (C) Constitutive-biasing, in which a constitutive meaning was hinted (Larry owns collections of Renaissance literature.), (N) Neutral context, which allowed both interpretations. Thirty-nine native speakers of English were asked to (i) rate each context-target sentence pair from a 1~5 scale (5=fully understandable), and (ii) choose possible interpretations for the target sentence given the context. The SI hypothesis predicts that comprehension is harder for the Neutral condition, as compared to the biasing conditions because no contextual information is provided to resolve an ambiguity. Also, comprehenders should obtain the specific interpretation corresponding to the context type. Results: (A) Agentive-biasing and (C) Constitutive-biasing were rated higher than (N) Neutral conditions (p< .001), while all conditions were within the acceptable range (> 3.5 on the 1~5 scale). This suggests that when lacking relevant contextual information, semantic ambiguity decreases comprehensibility. The interpretation task shows that the participants selected the biased agentive/constitutive reading for condition (A) and (C) respectively. For the Neutral condition, the agentive and constitutive readings were chosen equally often. Conclusion: These findings support the SI hypothesis: the meaning of AspV sentences is conceptualized as a parthood relation involving structured individuals. We argue that semantic representation makes reference to spatial structured-ness (abstracted axis). To obtain an appropriate interpretation, comprehenders utilize contextual information to enrich the conceptual representation of the sentence in question. This study connects semantic structure to human’s conceptual structure, and provides a processing model that incorporates contextual retrieval.

Keywords: ambiguity resolution, contextual retrieval, spatial structured-ness, structured individual

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Authors: Ankush, Harshit Sosan Lakra, Rachita Kuthial

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Climate change poses significant challenges to vulnerable communities, particularly indigenous populations in ecologically sensitive regions. Jharkhand, located in the heart of India, is home to several indigenous communities, including the Particularly Vulnerable Tribal Groups (PVTGs). The Indigenous architecture of the region functions as a significant reservoir of climate adaptation wisdom. It explores the architectural analysis encompassing the construction materials, construction techniques, design principles, climate responsiveness, cultural relevance, adaptation, integration with the environment and traditional wisdom that has evolved through generations, rooted in cultural and socioeconomic traditions, and has allowed these communities to thrive in a variety of climatic zones, including hot and dry, humid, and hilly terrains to withstand the test of time. Despite their historical resilience to adverse climatic conditions, PVTG tribal communities face new and amplified challenges due to the accelerating pace of climate change. There is a significant research void that exists in assimilating their traditional practices and local wisdom into contemporary climate resilience initiatives. Most of the studies place emphasis on technologically advanced solutions, often ignoring the invaluable Indigenous Local knowledge that can complement and enhance these efforts. This research gap highlights the need to bridge the disconnect between indigenous knowledge and contemporary climate adaptation strategies. The study aims to explore and leverage indigenous knowledge of vernacular architecture as a strategic tool for enhancing climatic resilience among PVTGs of the region. The first objective is to understand the traditional wisdom of vernacular architecture by analyzing and documenting distinct architectural practices and cultural significance of PVTG communities, emphasizing construction techniques, materials and spatial planning. The second objective is to develop culturally sensitive climatic resilience strategies based on findings of vernacular architecture by employing a multidisciplinary research approach that encompasses ethnographic fieldwork climate data assessment considering multiple variables such as temperature variations, precipitation patterns, extreme weather events and climate change reports. This will be a tailor-made solution integrating indigenous knowledge with modern technology and sustainable practices. With the involvement of indigenous communities in the process, the research aims to ensure that the developed strategies are practical, culturally appropriate, and accepted. To foster long-term resilience against the global issue of climate change, we can bridge the gap between present needs and future aspirations with Traditional wisdom, offering sustainable solutions that will empower PVTG communities. Moreover, the study emphasizes the significance of preserving and reviving traditional Architectural wisdom for enhancing climatic resilience. It also highlights the need for cooperative endeavors of communities, stakeholders, policymakers, and researchers to encourage integrating traditional Knowledge into Modern sustainable design methods. Through these efforts, this research will contribute not only to the well-being of PVTG communities but also to the broader global effort to build a more resilient and sustainable future. Also, the Indigenous communities like PVTG in the state of Jharkhand can achieve climatic resilience while respecting and safeguarding the cultural heritage and peculiar characteristics of its native population.

Keywords: vernacular architecture, climate change, resilience, PVTGs, Jharkhand, indigenous people, India

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Authors: Kaleem Kashif, Tayyaba Anam, Yizhi Wu

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This paper introduces a methodology for advancing Italian speech vowels landmark detection within the distinctive feature-based speech recognition domain. Leveraging the legacy tool 'xkl' by integrating combined convolutional neural networks (CNNs) and recurrent neural networks (RNNs), the study presents a comprehensive enhancement to the 'xkl' legacy software. This integration incorporates re-assigned spectrogram methodologies, enabling meticulous acoustic analysis. Simultaneously, our proposed model, integrating combined CNNs and RNNs, demonstrates unprecedented precision and robustness in landmark detection. The augmentation of re-assigned spectrogram fusion within the 'xkl' software signifies a meticulous advancement, particularly enhancing precision related to vowel formant estimation. This augmentation catalyzes unparalleled accuracy in landmark detection, resulting in a substantial performance leap compared to conventional methods. The proposed model emerges as a state-of-the-art solution in the distinctive feature-based speech recognition systems domain. In the realm of deep learning, a synergistic integration of combined CNNs and RNNs is introduced, endowed with specialized temporal embeddings, harnessing self-attention mechanisms, and positional embeddings. The proposed model allows it to excel in capturing intricate dependencies within Italian speech vowels, rendering it highly adaptable and sophisticated in the distinctive feature domain. Furthermore, our advanced temporal modeling approach employs Bayesian temporal encoding, refining the measurement of inter-landmark intervals. Comparative analysis against state-of-the-art models reveals a substantial improvement in accuracy, highlighting the robustness and efficacy of the proposed methodology. Upon rigorous testing on a database (LaMIT) speech recorded in a silent room by four Italian native speakers, the landmark detector demonstrates exceptional performance, achieving a 95% true detection rate and a 10% false detection rate. A majority of missed landmarks were observed in proximity to reduced vowels. These promising results underscore the robust identifiability of landmarks within the speech waveform, establishing the feasibility of employing a landmark detector as a front end in a speech recognition system. The synergistic integration of re-assigned spectrogram fusion, CNNs, RNNs, and Bayesian temporal encoding not only signifies a significant advancement in Italian speech vowels landmark detection but also positions the proposed model as a leader in the field. The model offers distinct advantages, including unparalleled accuracy, adaptability, and sophistication, marking a milestone in the intersection of deep learning and distinctive feature-based speech recognition. This work contributes to the broader scientific community by presenting a methodologically rigorous framework for enhancing landmark detection accuracy in Italian speech vowels. The integration of cutting-edge techniques establishes a foundation for future advancements in speech signal processing, emphasizing the potential of the proposed model in practical applications across various domains requiring robust speech recognition systems.

Keywords: landmark detection, acoustic analysis, convolutional neural network, recurrent neural network

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Authors: Adriana Muñoz-Talavera, Ana Rosa Rincón-Sánchez, Abraham Escobedo-Moratilla, María Cristina Islas-Carbajal, Miguel Ángel Gómez-Lim

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The highest rates of diabetes incidence and prevalence worldwide will increase the number of diabetic patients requiring insulin or insulin analogues. Then, current production systems would not be sufficient to meet the future market demands. Therefore, developing efficient expression systems for insulin and insulin analogues are needed. In addition, insulin analogues with better pharmacokinetics and pharmacodynamics properties and without mitogenic potential will be required. SCI-57 (single chain insulin-57) is an insulin analogue having 10 times greater affinity to the insulin receptor, higher resistance to thermal degradation than insulin, native mitogenicity and biological effect. Plants as expression platforms have been used to produce recombinant proteins because of their advantages such as cost-effectiveness, posttranslational modifications, absence of human pathogens and high quality. Immunoglobulin production with a yield of 50% has been achieved by transient expression in Nicotiana benthamiana (Nb). The aim of this study is to produce SCI-57 by transient expression in Nb. Methodology: DNA sequence encoding SCI-57 was cloned in pICH31070. This construction was introduced into Agrobacterium tumefaciens by electroporation. The resulting strain was used to infiltrate leaves of Nb. In order to isolate SCI-57, leaves from transformed plants were incubated 3 hours with the extraction buffer therefore filtrated to remove solid material. The resultant protein solution was subjected to anion exchange chromatography on an FPLC system and ultrafiltration to purify SCI-57. Detection of SCI-57 was made by electrophoresis pattern (SDS-PAGE). Protein band was digested with trypsin and the peptides were analyzed by Liquid chromatography tandem-mass spectrometry (LC-MS/MS). A purified protein sample (20µM) was analyzed by ESI-Q-TOF-MS to obtain the ionization pattern and the exact molecular weight determination. Chromatography pattern and impurities detection were performed using RP-HPLC using recombinant insulin as standard. The identity of the SCI-57 was confirmed by anti-insulin ELISA. The total soluble protein concentration was quantified by Bradford assay. Results: The expression cassette was verified by restriction mapping (5393 bp fragment). The SDS-PAGE of crude leaf extract (CLE) of transformed plants, revealed a protein of about 6.4 kDa, non-present in CLE of untransformed plants. The LC-MS/MS results displayed one peptide with a high score that matches SCI-57 amino acid sequence in the sample, confirming the identity of SCI-57. From the purified SCI-57 sample (PSCI-57) the most intense charge state was 1069 m/z (+6) on the displayed ionization pattern corresponding to the molecular weight of SCI-57 (6412.6554 Da). The RP-HPLC of the PSCI-57 shows the presence of a peak with similar retention time (rt) and UV spectroscopic profile to the insulin standard (SCI-57 rt=12.96 and insulin rt=12.70 min). The collected SCI-57 peak had ELISA signal. The total protein amount in CLE from transformed plants was higher compared to untransformed plants. Conclusions: Our results suggest the feasibility to produce insulin analogue SCI-57 by transient expression in Nicotiana benthamiana. Further work is being undertaken to evaluate the biological activity by glucose uptake by insulin-sensitive and insulin-resistant murine and human cultured adipocytes.

Keywords: insulin analogue, mass spectrometry, Nicotiana benthamiana, transient expression

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Authors: Batul Kagalwala

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The nervous system is made up of complex delicate structures such as the spinal cord, peripheral nerves and the brain. These are prone to various types of injury ranging from neurodegenerative diseases to trauma leading to diseases like Parkinson's, Alzheimer's, multiple sclerosis, amyotrophic lateral sclerosis (ALS), multiple system atrophy etc. Unfortunately, because of the complicated structure of nervous system, spontaneous regeneration, repair and healing is seldom seen due to which brain damage, peripheral nerve damage and paralysis from spinal cord injury are often permanent and incapacitating. Hence, innovative and standardized approach is required for advance treatment of neurological injury. Nigella sativa (N. sativa), an annual flowering plant native to regions of southern Europe and Asia; has been suggested to have neuroprotective and anti-seizures properties. Neuroregeneration is found to occur in damaged cells when treated using extract of N. sativa. Due to its proven health benefits, lots of experiments are being conducted to extract all the benefits from the plant. The flowers are delicate and are usually pale blue and white in color with small black seeds. These seeds are the source of active components such as 30–40% fixed oils, 0.5–1.5% essential oils, pharmacologically active components containing thymoquinone (TQ), ditimoquinone (DTQ) and nigellin. In traditional medicine, this herb was identified to have healing properties and was extensively used Middle East and Far East for treating diseases such as head ache, back pain, asthma, infections, dysentery, hypertension, obesity and gastrointestinal problems. Literature studies have confirmed the extract of N. sativa seeds and TQ have inhibitory effects on inducible nitric oxide synthase and production of nitric oxide as well as anti-inflammatory and anticancer activities. Experimental investigation will be conducted to understand which ingredient of N. sativa causes neuroregeneration and roots to its healing property. An aqueous/ alcoholic extract of N. sativa will be made. Seed oil is also found to have used by researchers to prepare such extracts. For the alcoholic extracts, the seeds need to be powdered and soaked in alcohol for a period of time and the alcohol must be evaporated using rotary evaporator. For aqueous extracts, the powder must be dissolved in distilled water to obtain a pure extract. The mobile phase will be the extract while the suitable stationary phase (substance that is a good adsorbent e.g. silica gels, alumina, cellulose etc.) will be selected. Different ingredients of N. sativa will be separated using High Performance Liquid Chromatography (HPLC) for treating damaged cells. Damaged brain cells will be treated individually and in different combinations of 2 or 3 compounds for different intervals of time. The most suitable compound or a combination of compounds for the regeneration of cells will be determined using DOE methodology. Later the gene will also be determined and using Polymerase Chain Reaction (PCR) it will be replicated in a plasmid vector. This plasmid vector shall be inserted in the brain of the organism used and replicated within. The gene insertion can also be done by the gene gun method. The gene in question can be coated on a micro bullet of tungsten and bombarded in the area of interest and gene replication and coding shall be studied. Investigation on whether the gene replicates in the organism or not will be examined.

Keywords: black cumin, brain cells, damage, extract, neuroregeneration, PCR, plasmids, vectors

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