Search results for: protein domains
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3185

Search results for: protein domains

1805 Inhibition of Glutamate Carboxypeptidase Activity Protects Retinal Ganglionic Cell Death Induced by Ischemia-Reperfusion by Reducing the Astroglial Activation in Rat

Authors: Dugeree Otgongerel, Kyong Jin Cho, Yu-Han Kim, Sangmee Ahn Jo

Abstract:

Excessive activation of glutamate receptor is thought to be involved in retinal ganglion cell (RGC) death after ischemia- reperfusion damage. Glutamate carboxypeptidase II (GCPII) is an enzyme responsible for the synthesis of glutamate. Several studies showed that inhibition of GCPII prevents or reduces cellular damage in brain diseases. Thus, in this study, we examined the expression of GCPII in rat retina and the role of GCPII in acute high IOP ischemia-reperfusion damage of eye by using a GCPII inhibitor, 2-(phosphonomethyl) pentanedioic acid (2-PMPA). Animal model of ischemia-reperfusion was induced by raising the intraocular pressure for 60 min and followed by reperfusion for 3 days. Rats were randomly divided into four groups: either intra-vitreous injection of 2-PMPA (11 or 110 ng per eye) or PBS after ischemia-reperfusion, 2-PMPA treatment without ischemia-reperfusion and sham-operated normal control. GCPII immunoreactivity in normal rat retina was detected weakly in retinal nerve fiber layer (RNFL) and retinal ganglionic cell layer (RGL) and also inner plexiform layer (IPL) and outer plexiform layer (OPL) strongly where are co-stained with an anti-GFAP antibody, suggesting that GCPII is expressed mostly in Muller and astrocytes. Immunostaining with anti-BRN antibody showed that ischemia- reperfusion caused RGC death (31.5 %) and decreased retinal thickness in all layers of damaged retina, but the treatment of 2-PMPA twice at 0 and 48 hour after reperfusion blocked these retinal damages. GCPII level in RNFL layer was enhanced after ischemia-reperfusion but was blocked by PMPA treatment. This result was confirmed by western blot analysis showing that the level of GCPII protein after ischemia- reperfusion increased by 2.2- fold compared to control, but this increase was blocked almost completely by 110 ng PMPA treatment. Interestingly, GFAP immunoreactivity in the retina after ischemia- reperfusion followed by treatment with PMPA showed similar pattern to GCPII, increase after ischemia-reperfusion but reduction to the normal level by PMPA treatment. Our data demonstrate that increase of GCPII protein level after ischemia-reperfusion injury is likely to cause glial activation and/or retinal cell death which are mediated by glutamate, and GCPII inhibitors may be useful in treatment of retinal disorders in which glutamate excitotoxicity is pathogenic.

Keywords: glutamate carboxypepptidase II, glutamate excitotoxicity, ischemia-reperfusion, retinal ganglion cell

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1804 The Effect of Pre-Cracks on Structural Strength of the Nextel Fibers: A Multiscale Modeling Approach

Authors: Seyed Mohammad Mahdi Zamani, Kamran Behdinan

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In this study, a multiscale framework is performed to model the strength of Nextel fibers in presence of an atomistic scale pre-crack at finite temperatures. The bridging cell method (BCM) is the multiscale technique applied in this study, which decomposes the system into the atomistic, bridging and continuum domains; solves the whole system in a finite element framework; and incorporates temperature dependent calculations. Since Nextel is known to be structurally stable and retain 70% of its initial strength up to 1100°C; simulations are conducted at both of the room temperatures, 25°C, and fire temperatures, 1200°C. Two cases are modeled for a pre-crack present in either phases of alumina or mullite of the Nextel structure. The materials’ response is studied with respect to deformation behavior and ultimate tensile strength. Results show different crack growth trends for the two cases, and as the temperature increases, the crack growth resistance and material’s strength decrease.

Keywords: Nextel fibers, multiscale modeling, pre-crack, ultimate tensile strength

Procedia PDF Downloads 419
1803 A New Block Cipher for Resource-Constrained Internet of Things Devices

Authors: Muhammad Rana, Quazi Mamun, Rafiqul Islam

Abstract:

In the Internet of Things (IoT), many devices are connected and accumulate a sheer amount of data. These Internet-driven raw data need to be transferred securely to the end-users via dependable networks. Consequently, the challenges of IoT security in various IoT domains are paramount. Cryptography is being applied to secure the networks for authentication, confidentiality, data integrity and access control. However, due to the resource constraint properties of IoT devices, the conventional cipher may not be suitable in all IoT networks. This paper designs a robust and effective lightweight cipher to secure the IoT environment and meet the resource-constrained nature of IoT devices. We also propose a symmetric and block-cipher based lightweight cryptographic algorithm. The proposed algorithm increases the complexity of the block cipher, maintaining the lowest computational requirements possible. The proposed algorithm efficiently constructs the key register updating technique, reduces the number of encryption rounds, and adds a new layer between the encryption and decryption processes.

Keywords: internet of things, cryptography block cipher, S-box, key management, security, network

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1802 Syrian-Armenian Women Refugees: Crossing Borders between the Past and the Present, Negotiating between the Private and the Public

Authors: Ani Kojoyan

Abstract:

The Syrian refugee crisis has been a matter of worldwide concern during the recent years. And though refugees’ problems are contextualized in terms of time and space, the refugee crisis still remains a global issue to discuss. Since the start of the conflict, Armenia has welcomed thousands of Syrian refugees too. Taking into consideration Armenia’s current socio-economic and geopolitical situation, the flow of refugees is a challenge both for the country and for refugees themselves. However, these people are not simply refugees from Syria, they are Syrian-Armenian refugees; people whose ancestors were survivals of the Armenian Genocide, perpetrated by the Ottoman Turks in 1915, people whose ancestors became refugees a century ago in Syria and now, ironically, a century later they follow their ancestors’ paths, turning into refugees themselves in their historical homeland, facing various difficulties, among them socio-economic, socio-ideological, and identity and gender issues, the latter being the main topic of discussion in the present paper. The situation presented above makes us discuss certain questions within this study: how do Syrian-Armenian refugees define themselves and their status? Which are their gender roles in the socio-economic context? How do social and economic challenges re-shape Syrian-Armenian women refugees’ identities? The study applies qualitative research methods of analysis, which includes semi-structured and in-depth interviews with 15 participants (18-25, 26-40 age groups), and two focus group works, involving 8 participants (18-35 age group) for each focus group activity. The activities were carried out in October 2016, Yerevan, Armenia. The study also includes Secondary Data Analysis. In addition, in order to centralize refugee women’s experiences and identity issues, the study adopts a qualitative lens from a feminist standpoint position. It is based on the assumption that human activity structures and limits understanding, and that the distorted comprehension of events or activities has emerged from the male-oriented dominant judgement which can be discovered through uncovering the understanding of the situation from women’s activity perspectives. The findings suggest that identity is dynamic, complex, over-changing and sensitive to time and space, gender and class. The process of re-shaping identity is even more complicated and multi-layered and is based on internal and external factors, conditioned by individual and collective needs and interests. Refugees are mostly considered as people who lost their identity in the past since they have no longer connection anywhere and try to find it in the present. In turn, female refugees, being a more vulnerable class, go through more complicated identity re-formulating discourse negotiations. They stand between the borders of the old and new, borders of lost and re-found selves, borders of creating and self-fashioning, between illusions and the challenging reality. Particularly, refugee women become more sensitive within the discourses of the private and the public domains: some of them try to create a ‘new-self’, creating their space in a new society, whereas others try to negotiate their affective/emotional labour within their own family domains.

Keywords: feminist standpoint position, gender, identity, refugee studies, Syrian-Armenian women refugees

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1801 Home Made Rice Beer Waste (Choak): A Low Cost Feed for Sustainable Poultry Production

Authors: Vinay Singh, Chandra Deo, Asit Chakrabarti, Lopamudra Sahoo, Mahak Singh, Rakesh Kumar, Dinesh Kumar, H. Bharati, Biswajit Das, V. K. Mishra

Abstract:

The most widely used feed resources in poultry feed, like maize and soybean, are expensive as well as in short supply. Hence, there is a need to utilize non-conventional feed ingredients to cut down feed costs. As an alternative, brewery by-products like brewers’ dried grains are potential non-conventional feed resources. North-East India is inhabited by many tribes, and most of these tribes prepare their indigenous local brew, mostly using rice grains as the primary substrate. Choak, a homemade rice beer waste, is an excellent and cheap source of protein and other nutrients. Fresh homemade rice beer waste (rice brewer’s grain) was collected locally. The proximate analysis indicated 28.53% crude protein, 92.76% dry matter, 5.02% ether extract, 7.83% crude fibre, 2.85% total ash, 0.67% acid insoluble ash, 0.91% calcium, and 0.55% total phosphorus. A feeding trial with 5 treatments (incorporating rice beer waste at the inclusion levels of 0,10,20,30 & 40% by replacing maize and soybean from basal diet) was conducted with 25 laying hens per treatment for 16 weeks under completely randomized design in order to study the production performance, blood-biochemical parameters, immunity, egg quality and cost economics of laying hens. The results showed substantial variations (P<0.01) in egg production, egg mass, FCR per dozen eggs, FCR per kg egg mass, and net FCR. However, there was not a substantial difference in either body weight or feed intake or in egg weight. Total serum cholesterol reduced significantly (P<0.01) at 40% inclusion of rice beer waste. Additionally, the egg haugh unit grew considerably (P<0.01) when the graded levels of rice beer waste increased. The inclusion of 20% rice brewers dried grain reduced feed cost per kg egg mass and per dozen egg production by Rs. 15.97 and 9.99, respectively. Choak (homemade rice beer waste) can thus be safely incorporated into the diet of laying hens at a 20% inclusion level for better production performance and cost-effectiveness.

Keywords: choak, rice beer waste, laying hen, production performance, cost economics

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1800 Telephone Health Service to Improve the Quality of Life of the People Living with AIDS in Eastern Nepal

Authors: Ram Sharan Mehta, Naveen Kumar Pandey, Binod Kumar Deo

Abstract:

Quality of Life (QOL) is an important component in the evaluation of the well-being of People Living with AIDS (PLWA). This study assessed the effectiveness of education intervention programme in improving the QOL of PLWA on ART attaining the ART-clinics at B. P. Koirala Institute of Health Sciences (BPKIHS), Nepal. A pre-experimental research design was used to conduct the study among the PLWA on ART at BPKIHS from June to August 2013 involving 60 PLWA on pre-test randomly. The mean age of the respondents was 36.70 ± 9.92, and majority of them (80%) were of age group of 25-50 years and Male (56.7%). After education intervention programme there is significant change in the QOL in all the four domains i.e. Physical (p=0.008), Psychological (p=0.019), Social (p=0.046) and Environmental (p=0.032) using student t-test at 0.05 level of significance. There is significant (p= 0.016) difference in the mean QOL scores of pre-test and post-test. High QOL scores in post-test after education intervention programme may reflective of the effectiveness of planned education interventions programme.

Keywords: telephone, AIDS, health service, Nepal

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1799 Efficient Layout-Aware Pretraining for Multimodal Form Understanding

Authors: Armineh Nourbakhsh, Sameena Shah, Carolyn Rose

Abstract:

Layout-aware language models have been used to create multimodal representations for documents that are in image form, achieving relatively high accuracy in document understanding tasks. However, the large number of parameters in the resulting models makes building and using them prohibitive without access to high-performing processing units with large memory capacity. We propose an alternative approach that can create efficient representations without the need for a neural visual backbone. This leads to an 80% reduction in the number of parameters compared to the smallest SOTA model, widely expanding applicability. In addition, our layout embeddings are pre-trained on spatial and visual cues alone and only fused with text embeddings in downstream tasks, which can facilitate applicability to low-resource of multi-lingual domains. Despite using 2.5% of training data, we show competitive performance on two form understanding tasks: semantic labeling and link prediction.

Keywords: layout understanding, form understanding, multimodal document understanding, bias-augmented attention

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1798 Usability Assessment of a Bluetooth-Enabled Resistance Exercise Band among Young Adults

Authors: Lillian M. Seo, Curtis L. Petersen, Ryan J. Halter, David Kotz, John A. Batsis

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Background: Resistance-based exercises effectively enhance muscle strength, which is especially important in older populations as it reduces the risk of disability. Our group developed a Bluetooth-enabled handle for resistance exercise bands that wirelessly transmits relative force data through low-energy Bluetooth to a local smartphone or similar device. The system has the potential to measure home-based exercise interventions, allowing health professionals to monitor compliance. Its feasibility has already been demonstrated in both clinical and field-based settings, but it remained unclear whether the system’s usability persisted upon repeated use. The current study sought to assess the usability of this system and its users’ satisfaction with repeated use by deploying the device among younger adults to gather formative information that can ultimately improve the device’s design for older adults. Methods: A usability study was conducted in which 32 participants used the above system. Participants executed 10 repetitions of four commonly performed exercises: bicep flexion, shoulder abduction, elbow extension, and triceps extension. Each completed three exercise sessions, separated by at least 24 hours to minimize muscle fatigue. At its conclusion, subjects completed an adapted version of the usefulness, satisfaction, and ease (USE) questionnaire – assessing the system across four domains: usability, satisfaction, ease of use, and ease of learning. The 20-item questionnaire examined how strongly a participant agrees with positive statements about the device on a seven-point Likert scale, with one representing ‘strongly disagree’ and seven representing ‘strongly agree.’ Participants’ data were aggregated to calculate mean response values for each question and domain, effectively assessing the device’s performance across different facets of the user experience. Summary force data were visualized using a custom web application. Finally, an optional prompt at the end of the questionnaire allowed for written comments and feedback from participants to elicit qualitative indicators of usability. Results: Of the n=32 participants, 13 (41%) were female; their mean age was 32.4 ± 11.8 years, and no participants had a physical impairment. No usability questions received a mean score < 5 of seven. The four domains’ mean scores were: usefulness 5.66 ± 0.35; satisfaction 6.23 ± 0.06; ease of use 6.25 ± 0.43; and ease of learning 6.50 ± 0.19. Representative quotes of the open-ended feedback include: ‘A non-rigid strap-style handle might be useful for some exercises,’ and, ‘Would need different bands for each exercise as they use different muscle groups with different strength levels.’ General impressions were favorable, supporting the expectation that the device would be a useful tool in exercise interventions. Conclusions: A simple usability assessment of a Bluetooth-enabled resistance exercise band supports a consistent and positive user experience among young adults. This study provides adequate formative data, assuring the next steps can be taken to continue testing and development for the target population of older adults.

Keywords: Bluetooth, exercise, mobile health, mHealth, usability

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1797 Dietary Diversification and Nutritional Education: A Strategy to Improve Child Food Security Status in the Rural Mozambique

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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1796 External Program Evaluation: Impacts and Changes on Government-Assisted Refugee Mothers

Authors: Akiko Ohta, Masahiro Minami, Yusra Qadir, Jennifer York

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The Home Instruction for Parents of Preschool Youngsters (HIPPY) is a home instruction program for mothers of children 3 to 5 years old. Using role-play as a method of teaching, the participating mothers work with their home visitors and learn how to deliver the HIPPY curriculum to their children. Applying HIPPY, Reviving Hope and Home for High-risk Refugee Mothers Program (RHH) was created to provide more personalized peer support and to respond to ongoing settlement challenges for isolated and vulnerable Government Assisted Refugee (GAR) mothers. GARs often have greater needs and vulnerabilities than other refugee groups. While the support is available, they often face various challenges and barriers in starting their new lives in Canada, such as inadequate housing, low first-language literacy levels, low competency in English or French, and social isolation. The pilot project was operated by Mothers Matter Centre (MMC) from January 2019 to March 2021 in partnership with the Immigrant Services Society of BC (ISSofBC). The formative evaluation was conducted by a research team at Simon Fraser University. In order to provide more suitable support for GAR mothers, RHH intended to offer more flexibility in HIPPY delivery, supported by a home visitor, to meet the need of refugee mothers facing various conditions and challenges; to have a pool of financial resources to be used for the RHH families when necessitated during the program period; to have another designated staff member, called a community navigator, assigned to facilitate the support system for the RHH families in their settlement; to have a portable device available for each RHH mother to navigate settlement support resources; and to provide other variations of the HIPPY curriculum as an option for the RHH mothers, including a curriculum targeting pre-HIPPY age children. Reflections on each program component was collected from RHH mothers and staff members of MMC and ISSofBC, including frontline workers and management staff, through individual interviews and focus group discussions. Each of the RHH program components was analyzed and evaluated by applying Moore’s four domains framework to identify key information and generate new knowledge (data). To capture RHH mothers’ program experience more in depth based on their own reflections, the photovoice method was used. Some photos taken by the mothers will be shared to illustrate their RHH experience as part of their life stories. Over the period of the program, this evaluation observed how RHH mothers became more confident in various domains, such as communicating with others, taking public transportations alone, and teaching their own child(ren). One of the major factors behind the success was their home visitors’ flexibility and creativity to create a more meaningful and tailored approach for each mother, depending on her background and personal situation. The role of the community navigator was tested out and improved during the program period. The community navigators took the key role to assess the needs of the RHH families and connect them with community resources. Both the home visitors and community navigators were immigrant mothers themselves and owing to their dedicated care for the RHH mothers; they were able to gain trust and work closely and efficiently with RHH mothers.

Keywords: refugee mothers, settlement support, program evaluation, Canada

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1795 LncRNA-miRNA-mRNA Networks Associated with BCR-ABL T315I Mutation in Chronic Myeloid Leukemia

Authors: Adenike Adesanya, Nonthaphat Wong, Xiang-Yun Lan, Shea Ping Yip, Chien-Ling Huang

Abstract:

Background: The most challenging mutation of the oncokinase BCR-ABL protein T315I, which is commonly known as the “gatekeeper” mutation and is notorious for its strong resistance to almost all tyrosine kinase inhibitors (TKIs), especially imatinib. Therefore, this study aims to identify T315I-dependent downstream microRNA (miRNA) pathways associated with drug resistance in chronic myeloid leukemia (CML) for prognostic and therapeutic purposes. Methods: T315I-carrying K562 cell clones (K562-T315I) were generated by the CRISPR-Cas9 system. Imatinib-treated K562-T315I cells were subjected to small RNA library preparation and next-generation sequencing. Putative lncRNA-miRNA-mRNA networks were analyzed with (i) DESeq2 to extract differentially expressed miRNAs, using Padj value of 0.05 as cut-off, (ii) STarMir to obtain potential miRNA response element (MRE) binding sites of selected miRNAs on lncRNA H19, (iii) miRDB, miRTarbase, and TargetScan to predict mRNA targets of selected miRNAs, (iv) IntaRNA to obtain putative interactions between H19 and the predicted mRNAs, (v) Cytoscape to visualize putative networks, and (vi) several pathway analysis platforms – Enrichr, PANTHER and ShinyGO for pathway enrichment analysis. Moreover, mitochondria isolation and transcript quantification were adopted to determine the new mechanism involved in T315I-mediated resistance of CML treatment. Results: Verification of the CRISPR-mediated mutagenesis with digital droplet PCR detected the mutation abundance of ≥80%. Further validation showed the viability of ≥90% by cell viability assay, and intense phosphorylated CRKL protein band being detected with no observable change for BCR-ABL and c-ABL protein expressions by Western blot. As reported by several investigations into hematological malignancies, we determined a 7-fold increase of H19 expression in K562-T315I cells. After imatinib treatment, a 9-fold increment was observed. DESeq2 revealed 171 miRNAs were differentially expressed K562-T315I, 112 out of these miRNAs were identified to have MRE binding regions on H19, and 26 out of the 112 miRNAs were significantly downregulated. Adopting the seed-sequence analysis of these identified miRNAs, we obtained 167 mRNAs. 6 hub miRNAs (hsa-let-7b-5p, hsa-let-7e-5p, hsa-miR-125a-5p, hsa-miR-129-5p, and hsa-miR-372-3p) and 25 predicted genes were identified after constructing hub miRNA-target gene network. These targets demonstrated putative interactions with H19 lncRNA and were mostly enriched in pathways related to cell proliferation, senescence, gene silencing, and pluripotency of stem cells. Further experimental findings have also shown the up-regulation of mitochondrial transcript and lncRNA MALAT1 contributing to the lncRNA-miRNA-mRNA networks induced by BCR-ABL T315I mutation. Conclusions: Our results have indicated that lncRNA-miRNA regulators play a crucial role not only in leukemogenesis but also in drug resistance, considering the significant dysregulation and interactions in the K562-T315I cell model generated by CRISPR-Cas9. In silico analysis has further shown that lncRNAs H19 and MALAT1 bear several complementary miRNA sites. This implies that they could serve as a sponge, hence sequestering the activity of the target miRNAs.

Keywords: chronic myeloid leukemia, imatinib resistance, lncRNA-miRNA-mRNA, T315I mutation

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1794 Inflammatory Changes Caused by Lipopolysaccharide in Odontoblasts

Authors: Virve Pääkkönen, Heidi M. Cuffaro, Leo Tjäderhane

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Objectives: Odontoblasts are the outermost cell layer of dental pulp and form the dentin. Importance of bacterial products, e.g. lipoteichoic acid (LTA), a cell wall component of Gram-positive bacteria and lipopolysaccharide (LPS), a cell wall component of Gram-negative bacteria, have been indicated in the pathogenesis of pulpitis. Gram-positive bacteria are more prevalent in superficial carious lesion while the amount gram-negative is higher in the deep lesions. Objective of this study was to investigate the effect of these bacterial products on inflammatory response of pulp tissue. Interleukins (IL) were of special interest. Various ILs have been observed in the dentin-pulp complex of carious tooth in vivo. Methods: Tissue culture method was used for testing the effect of LTA and LPS on human odontoblasts. Enzymatic isolation technique was used to extract living odontoblasts for cell cultures. DNA microarray and quantitative PCR (qPCR) were used to characterize the changes in the expression profile of the tissue cultured odontoblasts. Laser microdissection was used to cut healthy and affected dentin and odontoblast layer directly under carious lesion for experiments. Cytokine array detecting 80 inflammatory cytokines was used to analyze the protein content of conditioned culture media as well as dentin and odontoblasts from the carious teeth. Results: LPS caused increased gene expression IL-1α, and -8 and decrease of IL-1β, 12 , -15 and -16 after 1h treatment, while after 24h treatment decrease of IL-8, -11 and 23 mRNAs was observed. LTA treatment caused cell death in the tissue cultured odontoblasts but in in the cell culture but not in cell culture. Cytokine array revealed at least 2-fold down-regulation of IL-1β, -10 and -12 in response to LPS treatment. Cytokine array of odontoblasts of carious teeth, as well as LPS-treated tissue-cultured odontoblasts, revealed increased protein amounts of IL-16, epidermal growth factor (EGF), angiogenin and IGFBP-1 as well as decreased amount of fractalkine. In carious dentin, increased amount of IL-1β, EGF and fractalkine was observed, as well as decreased level of GRO-1 and HGF. Conclusion: LPS caused marked changes in the expression of inflammatory cytokines in odontoblasts. Similar changes were observed in the odontoblasts cut directly under the carious lesion. These results help to shed light on the inflammatory processes happening during caries.

Keywords: inflammation, interleukin, lipoteichoic acid, odontoblasts

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1793 Multimodal Data Fusion Techniques in Audiovisual Speech Recognition

Authors: Hadeer M. Sayed, Hesham E. El Deeb, Shereen A. Taie

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In the big data era, we are facing a diversity of datasets from different sources in different domains that describe a single life event. These datasets consist of multiple modalities, each of which has a different representation, distribution, scale, and density. Multimodal fusion is the concept of integrating information from multiple modalities in a joint representation with the goal of predicting an outcome through a classification task or regression task. In this paper, multimodal fusion techniques are classified into two main classes: model-agnostic techniques and model-based approaches. It provides a comprehensive study of recent research in each class and outlines the benefits and limitations of each of them. Furthermore, the audiovisual speech recognition task is expressed as a case study of multimodal data fusion approaches, and the open issues through the limitations of the current studies are presented. This paper can be considered a powerful guide for interested researchers in the field of multimodal data fusion and audiovisual speech recognition particularly.

Keywords: multimodal data, data fusion, audio-visual speech recognition, neural networks

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1792 The AI Arena: A Framework for Distributed Multi-Agent Reinforcement Learning

Authors: Edward W. Staley, Corban G. Rivera, Ashley J. Llorens

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Advances in reinforcement learning (RL) have resulted in recent breakthroughs in the application of artificial intelligence (AI) across many different domains. An emerging landscape of development environments is making powerful RL techniques more accessible for a growing community of researchers. However, most existing frameworks do not directly address the problem of learning in complex operating environments, such as dense urban settings or defense-related scenarios, that incorporate distributed, heterogeneous teams of agents. To help enable AI research for this important class of applications, we introduce the AI Arena: a scalable framework with flexible abstractions for distributed multi-agent reinforcement learning. The AI Arena extends the OpenAI Gym interface to allow greater flexibility in learning control policies across multiple agents with heterogeneous learning strategies and localized views of the environment. To illustrate the utility of our framework, we present experimental results that demonstrate performance gains due to a distributed multi-agent learning approach over commonly-used RL techniques in several different learning environments.

Keywords: reinforcement learning, multi-agent, deep learning, artificial intelligence

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1791 Polyclonal IgG glycosylation in Patients with Pediatric Appendicitis

Authors: Dalma Dojcsák, Csaba Váradi, Flóra Farkas, Tamás Farkas, János Papp, Béla Viskolcz

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Background: Appendicitis is a common acute inflammatory condition in both children and adults, but current laboratory markers such as C-reactive protein (CRP), white blood cell count (WBC), absolute neutrophil count (ANC), and red blood cell count (RNC) lack specificity in detecting appendicitis-related inflammation. N-glycosylation, an asparagine-linked glycosylation process, plays a vital role in cellular interactions, angiogenesis, immune response, and effector functions. Altered N-glycosylation impacts tumor growth and both acute and chronic inflammatory processes. IgG, the second most abundant glycoprotein in serum, shows altered glycosylation patterns during inflammation, suggesting that IgG glycan modifications may serve as potential biomarkers for appendicitis. Specifically, increased levels of agalactosylated IgG glycans are a known feature of various inflammatory conditions, potentially including appendicitis. Identifying pediatric appendicitis remains challenging due to the absence of specific biomarkers, which makes diagnosis reliant on clinical symptoms, imaging such as ultrasound, and nonspecific lab indicators (e.g., CRP, WBC, ANC). In this study, we analyzed the IgG derived N-glycome in pediatric patients with appendicitis compared with healthy controls. Methodology: The N-glycome was analyzed by high-performance liquid-chromatography combined with mass spectrometry. IgG was isolated from serum samples by Protein G column. The IgG derived glycans were released by enzymatic deglycosylation and fluorescent tags were attached to each glycan moiety, which made necessitates the sample clean-up for further reliable quantitation. Overall, 38 controls and 40 serum samples diagnosed with pediatric appendicitis were analyzed by HILIC-MS methods. Multivariate statistical tests were performed with area percentage under the peak data derived from the integrated peaks, which were obtained from the chromatograms. Conclusions: Our results represented the altered N-glycome of IgG in pediatric appendicitis is similar with other observations. The glycosylation pattern reported so far for IgG is characterized by decreased galactosylation and sialylation, and an increase in fucosylation.

Keywords: N-glycosylation, liquid chromatography, mass spectrometry, inflammation, appendicitis, immunoglobulin G

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1790 Unlocking the Health Benefits of Goat Meat

Authors: K. Makangali, G. Tokysheva, A. Shoman

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Goat meat and goat meat products have garnered increasing attention within the realm of nutrition and health due to their potential to provide a myriad of benefits. This scientific article presents a comprehensive review of the health advantages associated with goat meat consumption and the products derived from it. The paper explores the nutritional content of goat meat, highlighting its favorable composition in terms of protein, essential minerals, and amino acids. It delves into the intricate balance of macronutrients, with lower fat and cholesterol levels compared to other meats, making goat meat a desirable choice for individuals seeking healthier dietary options.

Keywords: goat meat, amino acid, nutrition, meat products, meat

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1789 Solanum tuberosum Ammonium Transporter Gene: Some Bioinformatics Insights

Authors: A. T. Adetunji, F. B. Lewu, R. Mundembe

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Plants require nitrogen (N) to support desired production levels. Nitrogen is available to plants in the form of nitrate or ammonium, which are transported into the cell with the aid of various transport proteins. Ammonium transporters (AMTs) play a role in the uptake of ammonium, the form in which nitrogen is preferentially absorbed by plants. Solanum tuberosum AMT1 (StAMT1) was characterized using molecular biology and bioinformatics methods. Nucleotide database sequences were used to design AMT1-specific primers which were used to amplify the AMT1 internal regions. Nucleotide sequencing, alignment and phylogenetic analysis assigned StAMT1 to the AMT1 family. The deduced amino acid sequences showed that StAMT1 is 92%, 83% and 76% similar to Solanum lycopersicum LeAMT1.1, Lotus japonicus LjAMT1.1 and Solanum lycopersicum LeAMT1.2 respectively. StAMT1 fragments were shown to correspond to the 5th - 10th trans-membrane domains. Residue StAMT1 D15 is predicted to be essential for ammonium transport, while mutations of StAMT1 S76A may further enhance ammonium transport.

Keywords: ammonium transporter, bioinformatics, nitrogen, primers, Solanum tuberosum

Procedia PDF Downloads 248
1788 Genome Analyses of Pseudomonas Fluorescens b29b from Coastal Kerala

Authors: Wael Ali Mohammed Hadi

Abstract:

Pseudomonas fluorescens B29B, which has asparaginase enzymatic activity, was isolated from the surface coastal seawater of Trivandrum, India. We report the complete Pseudomonas fluorescens B29B genome sequenced, identified, and annotated from a marine source. We find the genome at most minuscule a 7,331,508 bp single circular chromosome with a GC content of 62.19% and 6883 protein-coding genes. Three hundred forty subsystems were identified, including two predicted asparaginases from the genome analysis of P. fluorescens B29B for further investigation. This genome data will help further industrial biotechnology applications of proteins in general and asparaginase as a target.

Keywords: pseudomonas, marine, asparaginases, Kerala, whole-genome

Procedia PDF Downloads 214
1787 Rehabilitation of CP Using Pediatric Functional Independent Measure (WeeFIM) as Indicator Instruments Suitable for CP: Saudi's Perspective

Authors: Bara M. Yousef

Abstract:

Kingdome of Saudi Arabia (KSA). High numbers of traffic accidents with sever, moderate and mild level of impairments admits to Sultan bin Abdulaziz humanitarian city. Over a period of 4 months the city received 111 male and 79 female subjects with CP, who received 4-6 weeks of rehabilitation and using WeeFIM score to measure rehabilitation outcomes. WeeFIM measures and covers various domains, such as: self-care, mobility, locomotion, communication and other psycho-social aspects. Our findings shed the light on the fact that nearly 85% of people at admission got better after rehabilitation program services at individual sever moderate and mild and has arrange of (59 out of 128 WeeFIM score) and by the time of discharge they leave the city with better FIM score close to (72 out of 128 WeeFIM score) for the entire study sample. WeeFIM score is providing fair evidence to rehabilitation specialists to assess their outcomes. However there is a need to implement other instruments and compare it to WeeFIM in order to reach better outcomes at discharge level.

Keywords: Cerepral Palsy (CP), pediatric Functional Independent Measure (WeeFIM), rehabilitation, disability

Procedia PDF Downloads 226
1786 The Implementation of Self-Determination Theory on the Opportunities and Challenges for Blended E-Learning in Motivating Egyptian Logistics Learners

Authors: Aisha Noour, Nick Hubbard

Abstract:

Learner motivation is considered an important premise for the Blended e-Learning (BL) method. BL is an effective learning method in multiple domains, which opens several opportunities for its participants to engage in the learning environment. This research explores the learners’ perspective of BL according to the Self-Determination Theory (SDT). It identifies the opportunities and challenges for using the BL in Logistics Education (LE) in Egyptian Higher Education (HE). SDT is approached from different perspectives within the relationship between Intrinsic Motivation (IM), Extrinsic Motivation (EM) and Amotivation (AM). A self-administered face-to-face questionnaire was used to collect data from learners who were geographically widely spread around three colleges of International Transport and Logistics (CILTs) at the Arab Academy for Science, Technology and Maritime Transport (AAST&MT) in Egypt. Six hundred and sixteen undergraduates responded to a questionnaire survey. Respondents were drawn from three branches in Greater Cairo, Alexandria, and Port Said. The data analysis used was SPSS 22 and AMOS 18.

Keywords: intrinsic motivation, extrinsic motivation, amotivation, blended e-learning, Self Determination Theory

Procedia PDF Downloads 419
1785 Anti-Aging Effects of Two Agricultural Plant Extracts and Their Underlying Mechanism

Authors: Shwu-Ling Peng, Chiung-Man Tsai, Chia-Jui Weng

Abstract:

Chronic micro-inflammation is a hallmark of many aging-related neurodegenerative and metabolic syndrome-driven diseases. In high glucose (HG) environment, reactive oxygen species (ROS) is generated and the ROS induced inflammation, cytokines secretion, DNA damage, and cell cycle arrest to lead to cellular senescence. Water chestnut shell (WCS) is a plant hull which containing polyphenolic compounds and showed antioxidant and anticancer activities. Orchid, which containing a natural polysaccharide compound, possesses many physiological activities including anti-inflammatory and neuroprotective effects. These agricultural plants might be able to reduce oxidative stress and inflammation. This study was used HG-induced human normal dermal fibroblasts (HG-HNDFs) as an in vitro model to disclose the effects of water extract of Phalaenopsis orchid flower (WEPF) and ethanol extract of water chestnut shell (EEWCS) on the anti-aging and their underlying molecular mechanisms. The toxicity of extracts on human normal dermal fibroblasts (HNDFs) was determined by MTT method. The senescence of cells was assayed by β-galactosidase (SA-β-gal) kit. ROS and nitrate production was analyzed by Intracellular ROS contents and ELISA, respectively. Western blotting was used to detect the proteins in cells. The results showed that the exposure of HNDFs to HG (30 mM) for 72 h were caused cellular senescence and arrested cells at G0/G1 phase. Indeed, the treatment of HG-HNDFs with WEPF (200 μg/ml) and EEWCS (10 μg/ml) significantly released cell cycle arrest and promoted cell proliferation. The G1/S phase transition regulatory proteins such as protein retinoblastoma (pRb), p53, and p16ᴵᴺᴷ⁴ᵃ depressed by WEPF and EEWCS were also observed. Additionally, the treatment of WEPF and EEWCS increased the activity of HO-1 through upregulating Nrf2 as well as decreased the ROS and NO of HG-HNDFs. Therefore, the senescence marker protein-30 (SMP30) in cells was diminished. In conclusion, the WEPF and EEWCS might inhibit HG-induced aging of HNDFs by reducing oxidative stress and free radicals.

Keywords: agricultural plant extract, anti-aging, high glucose, Phalaenopsis orchid flower, water chestnut shell

Procedia PDF Downloads 154
1784 On Modules over Dedekind Prime Rings

Authors: Elvira Kusniyanti, Hanni Garminia, Pudji Astuti

Abstract:

This research studies an interconnection between finitely generated uniform modules and Dedekind prime rings. The characterization of modules over Dedekind prime rings that will be investigated is an adoption of Noetherian and hereditary concept. Dedekind prime rings are Noetherian and hereditary rings. This property of Dedekind prime rings is a background of the idea of adopting arises. In Noetherian area, it was known that a ring R is Noetherian ring if and only if every finitely generated R-module is a Noetherian module. Similar to that result, a characterization of the hereditary ring is related to its projective modules. That is, a ring R is hereditary ring if and only if every projective R-module is a hereditary module. Due to the above two results, we suppose that characterization of a Dedekind prime ring can be analyzed from finitely generated modules over it. We propose a conjecture: a ring R is a Dedekind prime ring if and only if every finitely generated uniform R-module is a Dedekind module. In this article, we will generalize a concept of the Dedekind module for non-commutative ring case and present a part of the above conjecture.

Keywords: dedekind domains, dedekind prime rings, dedekind modules, uniform modules

Procedia PDF Downloads 441
1783 Numerical Modeling on the Vehicle Interior Noise Produced by Rain-the-Roof Excitation

Authors: Zilong Peng, Jun Fan

Abstract:

With the improvement of the living standards, the requirement on the acoustic comfort of the vehicle interior environment is becoming higher. The rain-the-roof producing interior noise is a common phenomenon for the vehicle, which usually discourages the conversation, especially for the heavy rain. This paper presents some numerical results about the rain-the-roof noise. The impact of each water drop is modeled as a short pulse, and the excitation locations on the roof are generated randomly. The vehicle body is simplified to a box closed with some certain-thickness shells. According to the main frequency components of the rain excitation, the analyzing frequency range is divided as low, high and middle frequency domains, which makes the vehicle body are modeled using finite element method (FEM), statistical energy analysis (SEA) and hybrid FE-SEA method, respectively. Furthermore, the effect of spatial distribution density and size of the rain on the sound pressure level are also discussed. These results may provide a guide for designing a more silent vehicle in the special weather.

Keywords: rain-the-roof noise, vehicle, finite element method, statistical energy analysis

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1782 Provenance in Scholarly Publications: Introducing the provCite Ontology

Authors: Maria Joseph Israel, Ahmed Amer

Abstract:

Our work aims to broaden the application of provenance technology beyond its traditional domains of scientific workflow management and database systems by offering a general provenance framework to capture richer and extensible metadata in unstructured textual data sources such as literary texts, commentaries, translations, and digital humanities. Specifically, we demonstrate the feasibility of capturing and representing expressive provenance metadata, including more of the context for citing scholarly works (e.g., the authors’ explicit or inferred intentions at the time of developing his/her research content for publication), while also supporting subsequent augmentation with similar additional metadata (by third parties, be they human or automated). To better capture the nature and types of possible citations, in our proposed provenance scheme metaScribe, we extend standard provenance conceptual models to form our proposed provCite ontology. This provides a conceptual framework which can accurately capture and describe more of the functional and rhetorical properties of a citation than can be achieved with any current models.

Keywords: knowledge representation, provenance architecture, ontology, metadata, bibliographic citation, semantic web annotation

Procedia PDF Downloads 117
1781 Robust Medical Image Watermarking Using Frequency Domain and Least Significant Bits Algorithms

Authors: Volkan Kaya, Ersin Elbasi

Abstract:

Watermarking and stenography are getting importance recently because of copyright protection and authentication. In watermarking we embed stamp, logo, noise or image to multimedia elements such as image, video, audio, animation and text. There are several works have been done in watermarking for different purposes. In this research work, we used watermarking techniques to embed patient information into the medical magnetic resonance (MR) images. There are two methods have been used; frequency domain (Digital Wavelet Transform-DWT, Digital Cosine Transform-DCT, and Digital Fourier Transform-DFT) and spatial domain (Least Significant Bits-LSB) domain. Experimental results show that embedding in frequency domains resist against one type of attacks, and embedding in spatial domain is resist against another group of attacks. Peak Signal Noise Ratio (PSNR) and Similarity Ratio (SR) values are two measurement values for testing. These two values give very promising result for information hiding in medical MR images.

Keywords: watermarking, medical image, frequency domain, least significant bits, security

Procedia PDF Downloads 287
1780 HIV-1 Nef Mediates Host Invasion by Differential Expression of Alpha-Enolase

Authors: Reshu Saxena, R. K. Tripathi

Abstract:

HIV-1 transmission and spread involves significant host-virus interaction. Potential targets for prevention of HIV-1 lies at the site of mucosal barriers. Thus a better understanding of how HIV-1 infects target cells at such sites and lead their invasion is required, with prime focus on the host determinants regulating HIV-1 spread. HIV-1 Nef is important for viral infectivity and pathogenicity. It promotes HIV-1 replication, facilitating immune evasion by interacting with various host factors and altering cellular pathways via multiple protein-protein interactions. In this study nef was sequenced from HIV-1 patients, and showed specific mutations revealing sequence variability in nef. To explore the difference in Nef functionality based on sequence variability we have studied the effects of HIV-1 Nef in human SupT1 T cell line and (THP-1) monocyte-macrophage cell lines through proteomics approach. 2D-Gel Electrophoresis in control and Nef-transfected SupT1 cells demonstrated several differentially expressed proteins with significant modulation of alpha-enolase. Through further studies, effects of Nef on alpha-enolase regulation were found to be cell lineage-specific, being stimulatory in macrophages/monocytes, inhibitory in T cells and without effect in HEK-293 cells. Cell migration and invasion studies were employed to determine biological function affected by Nef mediated regulation of alpha-enolase. Cell invasion was enhanced in THP-1 cells but was inhibited in SupT1 cells by wildtype nef. In addition, the modulation of enolase and cell invasion remained unaffected by a unique nef variant. These results indicated that regulation of alpha-enolase expression and invasive property of host cells by Nef is sequence specific, suggesting involvement of a particular motif of Nef. To precisely determine this site, we designed a heptapeptide including the suggested alpha-enolase regulating sequence of nef and a nef mutant with deletion of this site. Macrophages/monocytes being the major cells affected by HIV-1 at mucosal barriers, were particularly investigated by the nef mutant and peptide. Both the nef mutant and heptapeptide led to inhibition of enhanced enolase expression and increased invasiveness in THP-1 cells. Together, these findings suggest a possible mechanism of host invasion by HIV-1 through Nef mediated regulation of alpha-enolase and identifies a potential therapeutic target for HIV-1 entry at mucosal barriers.

Keywords: HIV-1 Nef, nef variants, host-virus interaction, tissue invasion

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1779 Design of a Dietetic Food: Case of Lebanese Kishk

Authors: Henri El Zakhem, Dona Shalhoub, Elias Atallah, Jessica Koura

Abstract:

Due to the increase of demand on dietetic food and the need for more types of diet food, the production of dietetic food is increasing and improving. This demand on dietetic food has triggered us to study the market in which we found that Kishk (Lebanese dairy product) diet is not available. Production of a low fat product which is diet Kishk was our concern. A strategy was followed to choose the right idea that will satisfy the need of the market. The whole process was studied and explained thoroughly. The percentage of fat was found to be 32.52 % in regular Kishk and 3.84 % in the diet Kishk produced. The new product has the advantage to be high in protein, low in fat.

Keywords: design and industrialization, dietetic, diet Kishk, fat

Procedia PDF Downloads 373
1778 Management of Meskit (Prosopis juliflora) Tree in Oman: The Case of Using Meskit (Prosopis juliflora) Pods for Feeding Omani Sheep

Authors: S. Al-Khalasi, O. Mahgoub, H. Yaakub

Abstract:

This study evaluated the use of raw or processed Prosopis juliflora (Meskit) pods as a major ingredient in a formulated ration to provide an alternative non-conventional concentrate for livestock feeding in Oman. Dry Meskit pods were reduced to lengths of 0.5- 1.0 cm to ensure thorough mixing into three diets. Meskit pods were subjected to two types of treatments; roasting and soaking. They were roasted at 150оC for 30 minutes using a locally-made roasting device (40 kg barrel container rotated by electric motor and heated by flame gas cooker). Chopped pods were soaked in tap water for 24 hours and dried for 2 days under the sun with frequent turning. The Meskit-pod-based diets (MPBD) were formulated and pelleted from 500 g/kg ground Meskit pods, 240 g/kg wheat bran, 200 g/kg barley grain, 50 g/kg local dried sardines and 10 g/kg of salt. Twenty four 10 months-old intact Omani male lambs with average body weight of 27.3 kg (± 0.5 kg) were used in a feeding trial for 84 days. They were divided (on body weight basis) and allocated to four diet combination groups. These were: Rhodes grass hay (RGH) plus a general ruminant concentrate (GRC); RGH plus raw Meskit pods (RMP) based concentrate; RGH plus roasted Meskit pods (ROMP) based concentrate; RGH plus soaked Meskit pods (SMP) based concentrate Daily feed intakes and bi-weekly body weights were recorded. MPBD had higher contents of crude protein (CP), acid detergent fibre (ADF) and neutral detergent fibre (NDF) than the GRC. Animals fed various types of MPBD did not show signs of ill health. There was a significant effect of feeding ROMP on the performance of Omani sheep compared to RMP and SMP. The ROMP fed animals had similar performance to those fed the GRC in terms of feed intake, body weight gain and feed conversion ratio (FCR).This study indicated that roasted Meskit pods based diet may be used instead of the commercial concentrate for feeding Omani sheep without adverse effects on performance. It offers a cheap alternative source of protein and energy for feeding Omani sheep. Also, it might help in solving the spread impact of Meskit trees, maintain the ecosystem and helping in preserving the local tree species.

Keywords: growth, Meskit, Omani sheep, Prosopis juliflora

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1777 Impact of Different Rearing Diets on the Performance of Adult Mealworms Tenebrio molitor

Authors: Caroline Provost, Francois Dumont

Abstract:

Production of insects for human and animal consumption is an increasingly important activity in Canada. Protein production is more efficient and less harmful to the environment using insect rearing compared to the impact of traditional livestock, poultry and fish farms. Insects are rich in essential amino acids, essential fatty acids and trace elements. Thus, insect-based products could be used as a food supplement for livestock and domestic animals and may even find their way into the diets of high performing athletes or fine dining. Nevertheless, several parameters remain to be determined to ensure efficient and profitable production that meet the potential of these sectors. This project proposes to improve the production processes, rearing diets and processing methods for three species with valuable gastronomic and nutritional potential: the common mealworms (Tenebrio molitor), the small mealworm (Alphitobius diaperinus), and the giant mealworm (Zophobas morio). The general objective of the project is to acquire specific knowledge for mass rearing of insects dedicated to animal and human consumption in order to respond to current market opportunities and meet a growing demand for these products. Mass rearing of the three species of mealworm was produced to provide the individuals needed for the experiments. Mealworms eat flour from different cereals (e.g. wheat, barley, buckwheat). These cereals vary in their composition (protein, carbohydrates, fiber, vitamins, antioxidant, etc.), but also in their purchase cost. Seven different diets were compared to optimize the yield of the rearing. Diets were composed of cereal flour (e.g. wheat, barley) and were either mixed or left alone. Female fecundity, larvae mortality and growing curves were observed. Some flour diets have positive effects on female fecundity and larvae performance while each mealworm was found to have specific diet requirements. Trade-offs between mealworm performance and costs need to be considered. Experiments on the effect of flour composition on several parameters related to performance and nutritional and gastronomic value led to the identification of a more appropriate diet for each mealworm.

Keywords: mass rearing, mealworm, human consumption, diet

Procedia PDF Downloads 147
1776 On the Other Side of Shining Mercury: In Silico Prediction of Cold Stabilizing Mutations in Serine Endopeptidase from Bacillus lentus

Authors: Debamitra Chakravorty, Pratap K. Parida

Abstract:

Cold-adapted proteases enhance wash performance in low-temperature laundry resulting in a reduction in energy consumption and wear of textiles and are also used in the dehairing process in leather industries. Unfortunately, the possible drawbacks of using cold-adapted proteases are their instability at higher temperatures. Therefore, proteases with broad temperature stability are required. Unfortunately, wild-type cold-adapted proteases exhibit instability at higher temperatures and thus have low shelf lives. Therefore, attempts to engineer cold-adapted proteases by protein engineering were made previously by directed evolution and random mutagenesis. The lacuna is the time, capital, and labour involved to obtain these variants are very demanding and challenging. Therefore, rational engineering for cold stability without compromising an enzyme's optimum pH and temperature for activity is the current requirement. In this work, mutations were rationally designed with the aid of high throughput computational methodology of network analysis, evolutionary conservation scores, and molecular dynamics simulations for Savinase from Bacillus lentus with the intention of rendering the mutants cold stable without affecting their temperature and pH optimum for activity. Further, an attempt was made to incorporate a mutation in the most stable mutant rationally obtained by this method to introduce oxidative stability in the mutant. Such enzymes are desired in detergents with bleaching agents. In silico analysis by performing 300 ns molecular dynamics simulations at 5 different temperatures revealed that these three mutants were found to be better in cold stability compared to the wild type Savinase from Bacillus lentus. Conclusively, this work shows that cold adaptation without losing optimum temperature and pH stability and additionally stability from oxidative damage can be rationally designed by in silico enzyme engineering. The key findings of this work were first, the in silico data of H5 (cold stable savinase) used as a control in this work, corroborated with its reported wet lab temperature stability data. Secondly, three cold stable mutants of Savinase from Bacillus lentus were rationally identified. Lastly, a mutation which will stabilize savinase against oxidative damage was additionally identified.

Keywords: cold stability, molecular dynamics simulations, protein engineering, rational design

Procedia PDF Downloads 140