Search results for: in vitro culture

Authors: Katarzyna Bialik-Wąs, Klaudia Pluta, Dagmara Malina, Małgorzata Miastkowska

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In recent years, biocompatible hydrogels have been used more and more in medical applications, especially as modern dressings and drug delivery systems. The main goal of this research was the characteristics of bio-hybrid hydrogel materials incorporated with the nanocarrier-drug system, which enable the release in a gradual and prolonged manner, up to 7 days. Therefore, the use of such a combination will provide protection against mechanical damage and adequate hydration. The proposed bio-hybrid hydrogels are characterized by: transparency, biocompatibility, good mechanical strength, and the dual release system, which allows for gradual delivery of the active substance, even up to 7 days. Bio-hybrid hydrogels based on sodium alginate (SA), poly(vinyl alcohol) (PVA), glycerine, and Aloe vera solution (AV) were obtained through the chemical crosslinking method using poly(ethylene glycol) diacrylate as a crosslinking agent. Additionally, a nanocarrier-drug system was incorporated into SA/PVA/AV hydrogel matrix. Here, studies were focused on the release profiles of active substances from bio-hybrid hydrogels using the USP4 method (DZF II Flow-Through System, Erweka GmbH, Langen, Germany). The equipment incorporated seven in-line flow-through diffusion cells. The membrane was placed over support with an orifice of 1,5 cm in diameter (diffusional area, 1.766 cm²). All the cells were placed in a cell warmer connected with the Erweka heater DH 2000i and the Erweka piston pump HKP 720. The piston pump transports the receptor fluid via seven channels to the flow-through cells and automatically adapts the setting of the flow rate. All volumes were measured by gravimetric methods by filling the chambers with Milli-Q water and assuming a density of 1 g/ml. All the determinations were made in triplicate for each cell. The release study of the model active substance was carried out using a regenerated cellulose membrane Spectra/Por®Dialysis Membrane MWCO 6-8,000 Carl Roth® Company. These tests were conducted in buffer solutions – PBS at pH 7.4. A flow rate of receptor fluid of about 4 ml /1 min was selected. The experiments were carried out for 7 days at a temperature of 37°C. The released concentration of the model drug in the receptor solution was analyzed using UV-Vis spectroscopy (Perkin Elmer Company). Additionally, the following properties of the modified materials were studied: physicochemical, structural (FT-IR analysis), morphological (SEM analysis). Finally, the cytotoxicity tests using in vitro method were conducted. The obtained results exhibited that the dual release system allows for the gradual and prolonged delivery of the active substances, even up to 7 days.

Keywords: wound dressings, SA/PVA hydrogels, nanocarrier-drug system, USP4 method

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Authors: Kgaugelo Lekota, Ayesha Hassim, Henriette Van Heerden

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Bacillus anthracis is the causative agent of anthrax that is composed of three genetic groups, namely A, B, and C. Clade-A is distributed world-wide, while sub-clades B has been identified in Kruger National Park (KNP), South Africa. KNP is one of the endemic anthrax regions in South Africa with distinctive genetic diversity. Genomic surveillance of KNP B. anthracis strains was employed on the historical culture collection isolates (n=67) dated from the 1990’s to 2015 using a whole genome sequencing approach. Whole genome single nucleotide polymorphism (SNPs) and pan-genomics analysis were used to define the B. anthracis genetic population structure. This study showed that KNP has heterologous B. anthracis strains grouping in the A-clade with more prominent ABr.005/006 (Ancient A) SNP lineage. The 2012 and 2015 anthrax isolates are dispersed amongst minor sub-clades that prevail in non-stabilized genetic evolution strains. This was augmented with non-parsimony informative SNPs of the B. anthracis strains across minor sub-clades of the Ancient A clade. Pan-genomics of B. anthracis showed a clear distinction between A and B-clade genomes with 11 374 predicted clusters of protein coding genes. Unique accessory genes of B-clade genomes that included biosynthetic cell wall genes and multidrug resistant of Fosfomycin. South Africa consists of diverse B. anthracis strains with unique defined SNPs. The sequenced B. anthracis strains in this study will serve as a means to further trace the dissemination of B. anthracis outbreaks globally and especially in South Africa.

Keywords: bacillus anthracis, whole genome single nucleotide polymorphisms, pangenomics, kruger national park

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Authors: Zahra Karimian, Effat Merghati Khoei, Raziyeh Maasoumi

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Background: Women's sexual well-being is center of focus in the field of sexology. Study of sexual behavior and investigating its predictors is important in women's health promotion. Objectives: This study aimed to explore the components of sexual behaviors and their possible associations with the women's demographic. Methods: A National Sexual Behavior Assessment Questionnaire was administered to 500 women ages 15 to 45 who referred to the public health centers seeking for health care services. The associations with demographic were examined. Results: From all participant, 31.8% of women obtain high score in the sexual capacity 21.2% in sexual motivation and 0.2% in sexual function. In sexual script component, 86.2% of women were holding traditional beliefs toward sexual behaviors; the majority (91.5%) of women believed in mutual and relational sexuality, 83.4% believed in androcentricity (male-dominated sexuality). Pearson correlation test showed significant positive correlations between sexual capacity, motivation, function and sexual script (p < 0.05). Regression model showed that sexual capacity is associated with women's education, age of her spouse. Sexual function and sexual motivation were significantly associated with the age of subjects' spouses. Conclusion: In this study, lower score was found in sexual performance while women were scored higher in the sexual capacity and motivation. We argue that these lower score in sexual performance more likely is due to the level of participants' religiosity and formation of their sexuality through an androcentric culture. Women's level of education and the spouse age appear to be predicting factors in the scores the subjects gained. We suggest that gender-specific and culturally sensitive sexuality education should be focus of women's health programs in Iran.

Keywords: sexual behaviors, women, health, Iran

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Authors: I. C. Orabueze, A. A. Amudalat, S. A. Adesegun, A. A. Usman

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Dental and associated oral diseases are increasingly affecting a considerable portion of the population and are considered some of the major causes of tooth loss, discomfort, mouth odor and loss of confidence. This study focused on the ethnobotanical survey of medicinal plants used in oral therapy and evaluation of the antimicrobial activities of methanolic extracts of two selected plants from the survey for their efficacy against dental microorganisms. The ethnobotanical survey was carried out in six herbal markets in Lagos State, Nigeria by oral interviewing and information obtained from an old family manually complied herbal medication book. Methanolic extracts of Olax subscorpioidea (stem bark) and Bridelia ferruginea (stem bark) were assayed for their antimicrobial activities against clinical oral isolates (Aspergillus fumigatus, Candida albicans, Streptococcus spp, Staphylococcus aureus, Lactobacillus acidophilus and Pseudomonas aeruginosa). In vitro microbial technique (agar well diffusion method and minimum inhibitory concentration (MIC) assay) were employed for the assay. Chlorhexidine gluconate was used as the reference drug for comparison with the extract results. And the preliminary phytochemical screening of the constituents of the plants were done. The ethnobotanical survey produced plants (28) of diverse family. Different parts of plants (seed, fruit, leaf, root, bark) were mentioned but 60% mentioned were either the stem or the bark. O. subscorpioidea showed considerable antifungal activity with zone of inhibition ranging from 2.650 – 2.000 cm against Aspergillus fumigatus but no such encouraging inhibitory activity was observed in the other assayed organisms. B. ferruginea showed antibacterial sensitivity against Streptococcus spp, Staphylococcus aureus, Lactobacillus acidophilus and Pseudomonas aeruginosa with zone of inhibitions ranging from 3.400 - 2.500, 2.250 - 1.600, 2.700 - 1.950, 2.225 – 1.525 cm respectively. The minimum inhibitory concentration of O. subscorpioidea against Aspergillus fumigatus was 51.2 mg ml-1 while that of B. ferruginea against Streptococcus spp was 0.1mg ml-1 and for Staphylococcus aureus, Lactobacillus acidophilus and Pseudomonas aeruginosa were 25.6 mg ml-1. A phytochemical analysis reveals the presence of alkaloids, saponins, cardiac glycoside, tannins, phenols and terpenoids in both plants, with steroids only in B. ferruginea. No toxicity was observed among mice given the two methanolic extracts (1000 mg Kg-1) after 21 days. The barks of both plants exhibited antimicrobial properties against periodontal diseases causing organisms assayed, thus up-holding their folkloric use in oral disorder management. Further research could be done viewing these extracts as combination therapy, checking for possible synergistic value in toothpaste and oral rinse formulations for reducing oral bacterial flora and fungi load.

Keywords: antimicrobial activities, Bridelia ferruginea, dental disinfection, methanolic extract, Olax subscorpioidea, ethnobotanical survey

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Authors: Ayesha Khatun

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In 21st century where markets are unstable, technologies rapidly proliferate, competitors multiply, products and services become obsolete almost overnight and customers demand low cost high value product, leveraging and harnessing knowledge is not just a potential source of competitive advantage rather a necessity in technology based and information intensive industries. Knowledge management focuses on leveraging the available knowledge and sharing the same among the individuals in the organization so that the employees can make best use of it towards achieving the organizational goals. Knowledge is not a discrete object. It is embedded in people and so difficult to transfer outside the immediate context that it becomes a major competitive advantage. However, internal transfer of knowledge among the employees is essential to maximize the use of knowledge available in the organization in an unstructured manner. But as knowledge is the source of competitive advantage for the organization it is also the source of competitive advantage for the individuals. People think that knowledge is power and sharing the same may lead to lose the competitive position. Moreover, the very nature of tacit knowledge poses many difficulties in sharing the same. But sharing tacit knowledge is the vital part of knowledge management process because it is the tacit knowledge which is inimitable. Knowledge management has been made synonymous with the use of software and technology leading to the management of explicit knowledge only ignoring personal interaction and forming of informal networks which are considered as the most successful means of sharing tacit knowledge. Factors responsible for effective sharing of tacit knowledge are grouped into –individual, organizational and technological factors. Different factors under each category have been identified. Creating a positive organizational culture, encouraging personal interaction, practicing reward system are some of the strategies that can help to overcome many of the barriers to effective sharing of tacit knowledge. Methodology applied here is completely secondary. Extensive review of relevant literature has been undertaken for the purpose.

Keywords: knowledge, tacit knowledge, knowledge management, sustainable competitive advantage, organization, knowledge sharing

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Authors: Dian Mayasari, Yosi Bayu Murti, Sylvia Utami Tunjung Pratiwi, Sudarsono

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Melastoma malabathricum L. is an Indo-Pacific herb that has been traditionally used to treat several ailments such as wounds, dysentery, diarrhea, toothache, and diabetes. This plant is common across tropical Indo-Pacific archipelagos and is tolerant of a range of soils, from low-lying areas subject to saltwater inundation to the salt-free conditions of mountain slopes. How the soil and environmental variation influences secondary metabolite production in the herb, and an understanding of the plant’s utility as traditional medicine, remain largely unknown and unexplored. The objective of this study is to evaluate the variability of the metabolic profiles of M. malabathricum L. across its geographic distribution. By employing high-performance liquid chromatography-diode array detector (HPLC-DAD), a highly established, simple, sensitive, and reliable method was employed for establishing the chemical fingerprints of 72 samples of M. malabathricum L. leaves from various geographical locations in Indonesia. Specimens collected from six terrestrial and archipelago regions of Indonesia were analyzed by HPLC to generate chromatogram peak profiles that could be compared across each region. Data corresponding to the common peak areas of HPLC chromatographic fingerprint were analyzed by hierarchical component analysis (HCA) and principal component analysis (PCA) to extract information on the most significant variables contributing to characterization and classification of analyzed samples data. Principal component values were identified as PC1 and PC2 with 41.14% and 19.32%, respectively. Based on variety and origin, the high-performance liquid chromatography method validated the chemical fingerprint results used to screen the in vitro antioxidant activity of M. malabathricum L. The result shows that the developed method has potential values for the quality of similar M. malabathrium L. samples. These findings provide a pathway for the development and utilization of references for the identification of M. malabathricum L. Our results indicate the importance of considering geographic distribution during field-collection efforts as they demonstrate regional metabolic variation in secondary metabolites of M. malabathricum L., as illustrated by HPLC chromatogram peaks and their antioxidant activities. The results also confirm the utility of this simple approach to a rapid evaluation of metabolic variation between plants and their potential ethnobotanical properties, potentially due to the environments from whence they were collected. This information will facilitate the optimization of growth conditions to suit particular medicinal qualities.

Keywords: fingerprint, high performance liquid chromatography, Melastoma malabathricum l., metabolic profiles, principal component analysis

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Authors: Z. Abram, V. Nadasan, J. Balint, J. L. Ferencz

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Introduction: Online smoking prevention programs became popular in the last time. In order to extend the use of such programs, existing applications can be adapted and translated in the native languages of the target groups. It is the first time that in Romania such a software was implemented. Our goal was to provide a computer-aided intervention with attractive content targeting high school students who are familiar with information and communication technology. Material and methods: ASPIRA is the Romanian/Hungarian adapted version of a smoking prevention program created in USA. Prior to apply the questionnaire and ASPIRA online program which contains five modules that include tests, videos and interactive games, the program was tested in some IT laboratories on a group of schoolchildren and students. The pilot study questionnaires were completed considering the opinions of young people and the functionality of the software. Results: Above 90% of participants reported a good or very good impression about the ASPIRA program. Only a small minority found that the program included some parts which were too long or reported the existence of any technical problems regarding the functionality of the software. 76% of the participants had little or very little difficulty in understanding the messages presented by the English speaking characters. Only 7.5% of the participants thought that the program included content that was not appropriate for the local culture. Conclusions: The vast majority of students reported favorite impressions about ASPIRA online program. High school students and boys were more critical. Language and cultural barriers did not have the potential to reduce in a significant manner the effectiveness of the tested program.

Keywords: smoking prevention, ASPIRA online program, youth opinions, language/cultural barriers

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Authors: Tang Mui Joo, Chan Eang Teng

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The use of social media has become more defined. It has been widely used for the purpose of business. More marketers are now using social media as tools to enhance their businesses. Whereas on the other hand, there are more and more people spending their time through mobile apps to be engaged in the social media sites like YouTube, Facebook, Twitter and others. Social media has even become common in Public Relations (PR). It has become number one platform for creating and sharing content. In view to this, social media has changed the rules in PR where it brings new challenges and opportunities to the profession. Although corporate websites, chat-rooms, email customer response facilities and electronic news release distribution are now viewed as standard aspects of PR practice, many PR practitioners are still struggling with the impact of new media though the implementation of social media is potentially reducing the cost of communication. It is to the point that PR practitioners are not fully embracing new media, they are ill-equipped to do so and they have a fear of the technology. Somehow that social media has become a new style of communication that is characterized by conversation and community. It has become a platform that allows individuals to interact with one another and build relationship among each other. Therefore, in the use of business world, consumers are able to interact with those companies that have joined any social media. Based on their experiences with social networking site interactions, they are also exposed to personal interaction while communicating. This paper is to study the impact of social media to PR. This paper discovers the potential changes of PR practices in a developing country like Malaysia. Eventually the study reflects on how PR practitioners are actually using social media in the country. This paper is based on two theories in its development of this research foundation. Media Ecology Theory is to support the impact and changes to PR. Social Penetration Theory is to reflect on how the use of social media is among PRs. This research is using survey with PR practitioners in its data collection. The results have shown that PR professionals value social media more than they actually use it and the way of organizations communicate had been changed due to the transformation of social media.

Keywords: new media, social media, PR, change of trend, communication, digital culture

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Authors: Saulutė Juzelėnienė, Skirmantė Šarkauskienė

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In this paper, it is aimed to analyse the multimodal representations of the concepts of LOVE and PASSION in Lithuanian graphic novel “Gertrūda”, by Gerda Jord. The research is based on the earlier findings by Forceville (2005), Eerden (2009) as well as insights made by Shihara and Matsunaka (2009) and Kövecses (2000). The domains of target and source of LOVE and PASSION metaphors in comics are expressed by verbal and non-verbal cues. The analysis of non-verbal cues adopts the concepts of rune and indexes. A pictorial rune is a graphic representation of an object that does not exist in reality in comics, such as lines, dashes, text "balloons", and pictorial index – a graphically represented object of reality, a real symptom expressing a certain emotion, such as a wide smile, furrowed eyebrows, etc. Indexes are often hyperbolized in comics. The research revealed that most frequent source domains are CLOSINESS/UNITY, NATURAL/ PHYSICAL FORCE, VALUABLE OBJECT, PRESSURE. The target is the emotion of LOVE/PASSION which belongs to a more abstract domain of psychological experience. In this kind of metaphor, the picture can be interpreted as representing the emotion of happiness. Data are taken from Lithuanian comic books and Internet sites, where comics have been presented. The data and the analysis we are providing in this article aims to reveal that there are pictorial metaphors that manifest conceptual metaphors that are also expressed verbally and that methodological framework constructed for the analysis in the papers by Forceville at all is applicable to other emotions and culture specific pictorial manifestations.

Keywords: multimodal metaphor, conceptual metaphor, comics, graphic novel, concept of love/passion

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Authors: Diobein C. Flores

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The populace in Municipality of Talaingod is composed of Ata-Manobo. The said lumads enrich their culture, orientation and self because the place is a hive of their tribe. In lieu, the study would analyze the participation of the Ata-Manobo in the making of school improvement plan (SIP). Thus, it recommends alternative policy options that would help strengthen their involvement. The school stakeholders-Ata Manobo representatives from students, parent-teacher association, alumni, basic sector, municipal/barangay government unit, civic/social organizations and other government various agencies are the key participants in this study. The research used descriptive design. The responses of the representatives were analyzed through the criteria involved in employing Rational Model. The technical dimension, administrative, political acceptability and economic are the criteria in revealing decision. The policy alternative option 3- recommends to formulate policy for the purpose of capacitating stakeholders or governing council members in the making of SIP was pointed out as the most preferred option. This could strengthen the participation among Ata-Manobo as stakeholders in planning. Hence, the formulation alternative policy- capacitating stakeholders in the crafting of school improvement plan is recommended. The suggested initiative would assist the Department of Education in forging consensus across neighborhoods during the making of SIP. The appropriation of the definite budget to be used during the conduct of capability building activities is also suggested. Training-workshops are identified as possible intervention to ensure that the stakeholders are equipped with necessary knowledge and skills needed in the making of SIP. Indeed, the equal opportunities for all stakeholders regardless of their life circumstances must be noted. With the belief, people must be empowered to take advantage and spearhead progress in the making of SIP.

Keywords: Ata-Manobo Tribe, stakeholders, school improvement plan, Municipality of Talaingod, Philippines

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Authors: Lamzira Ebralidze, Aleksandre Tsertsvadze, Dali Berashvili, Aliosha Bakuridze

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Nowadays, there is an increasing demand for biologically active substances from vegetable, animal, and mineral resources. In terms of the use of natural compounds, pharmaceutical, cosmetic, and nutrition industry has big interest. The biggest drawback of conventional extraction methods is the need to use a large volume of organic extragents. The removal of the organic solvent is a multi-stage process. And their absolute removal cannot be achieved, and they still appear in the final product as impurities. A large amount of waste containing organic solvent damages not only human health but also has the harmful effects of the environment. Accordingly, researchers are focused on improving the extraction methods, which aims to minimize the use of organic solvents and energy sources, using alternate solvents and renewable raw materials. In this context, green extraction principles were formed. Green Extraction is a need of today’s environment. Green Extraction is the concept, and it totally corresponds to the challenges of the 21st century. The extraction of biologically active compounds based on green extraction principles is vital from the view of preservation and maintaining biodiversity. Novel technologies of green extraction are known, such as "cold methods" because during the extraction process, the temperature is relatively lower, and it doesn’t have a negative impact on the stability of plant compounds. Novel technologies provide great opportunities to reduce or replace the use of organic toxic solvents, the efficiency of the process, enhance excretion yield, and improve the quality of the final product. The objective of the research is the development of green technologies of flavonoids containing preparations. Methodology: At the first stage of the research, flavonoids containing preparations (Tincture Herba Leonuri, flamine, rutine) were prepared based on conventional extraction methods: maceration, bismaceration, percolation, repercolation. At the same time, the same preparations were prepared based on green technologies, microwave-assisted, UV extraction methods. Product quality characteristics were evaluated by pharmacopeia methods. At the next stage of the research technological - economic characteristics and cost efficiency of products prepared based on conventional and novel technologies were determined. For the extraction of flavonoids, water is used as extragent. Surface-active substances are used as co-solvent in order to reduce surface tension, which significantly increases the solubility of polyphenols in water. Different concentrations of water-glycerol mixture, cyclodextrin, ionic solvent were used for the extraction process. In vitro antioxidant activity will be studied by the spectrophotometric method, using DPPH (2,2-diphenyl-1- picrylhydrazyl) as an antioxidant assay. The advantage of green extraction methods is also the possibility of obtaining higher yield in case of low temperature, limitation extraction process of undesirable compounds. That is especially important for the extraction of thermosensitive compounds and maintaining their stability.

Keywords: extraction, green technologies, natural resources, flavonoids

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Authors: Hsiao-Chuan Huang, King-Lung Kuo, Mei-Hsin Lo, Hsiao-Yun Chou, Yusen Lin

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The most robust and economical method for laboratory diagnosis of TB is to identify mycobacterial bacilli (AFB) under acid-fast staining despite its disadvantages of low sensitivity and labor-intensive. Though digital pathology becomes popular in medicine, an automated microscopic system for microbiology is still not available. A new AI-assisted automated microscopic system, consisting of a microscopic scanner and recognition program powered by big data and deep learning, may significantly increase the sensitivity of TB smear microscopy. Thus, the objective is to evaluate such an automatic system for the identification of AFB. A total of 5,930 smears was enrolled for this study. An intelligent microscope system (TB-Scan, Wellgen Medical, Taiwan) was used for microscopic image scanning and AFB detection. 272 AFB smears were used for transfer learning to increase the accuracy. Referee medical technicians were used as Gold Standard for result discrepancy. Results showed that, under a total of 1726 AFB smears, the automated system's accuracy, sensitivity and specificity were 95.6% (1,650/1,726), 87.7% (57/65), and 95.9% (1,593/1,661), respectively. Compared to culture, the sensitivity for human technicians was only 33.8% (38/142); however, the automated system can achieve 74.6% (106/142), which is significantly higher than human technicians, and this is the first of such an automated microscope system for TB smear testing in a controlled trial. This automated system could achieve higher TB smear sensitivity and laboratory efficiency and may complement molecular methods (eg. GeneXpert) to reduce the total cost for TB control. Furthermore, such an automated system is capable of remote access by the internet and can be deployed in the area with limited medical resources.

Keywords: TB smears, automated microscope, artificial intelligence, medical imaging

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Authors: P.P. Rupasinghe, A.H. Farid

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The Aleutian mink disease virus (AMDV, Carnivore amdoparvovirus 1) causes persistent infection, plasmacytosis, and formation and deposition of immune complexes in various organs in adult mink, leading to glomerulonephritis, arteritis and sometimes death. The disease has no cure nor an effective vaccine, and identification and culling of mink positive for anti-AMDV antibodies have not been successful in controlling the infection in many countries. The failure to eradicate the virus from infected farms may be caused by keeping false-negative individuals on the farm, virus transmission from wild animals, or neighboring farms. The identification of sources of infection, which can be performed by comparing viral sequences, is important in the success of viral eradication programs. High mutation rates could cause inaccuracies when viral sequences are used to trace back an infection to its origin. There is no published information on the mutation rate of AMDV either in vivo or in vitro. The in vivo estimation is the most accurate method, but it is difficult to perform because of the inherent technical complexities, namely infecting live animals, the unknown numbers of viral generations (i.e., infection cycles), the removal of deleterious mutations over time and genetic drift. The objective of this study was to determine the mutation rate of AMDV on which no information was available. A homogenate was prepared from the spleen of one naturally infected American mink (Neovison vison) from Nova Scotia, Canada (parental template). The near full-length genome of this isolate (91.6%, 4,143 bp) was bidirectionally sequenced. A group of black mink was inoculated with this homogenate (descendant mink). Spleen sampled were collected from 10 descendant mink after 16 weeks post-inoculation (wpi) and from anther 10 mink after 176 wpi, and their near-full length genomes were bi-directionally sequenced. Sequences of these mink were compared with each other and with the sequence of the parental template. The number of nucleotide substitutions at 176 wpi was 3.1 times greater than that at 16 wpi (113 vs 36) whereas the estimates of mutation rate at 176 wpi was 3.1 times lower than that at 176 wpi (2.85×10-3 vs 9.13×10-4 substitutions/ site/ year), showing a decreasing trend in the mutation rate per unit of time. Although there is no report on in vivo estimate of the mutation rate of DNA viruses in animals using the same method which was used in the current study, these estimates are at the higher range of reported values for DNA viruses determined by various techniques. These high estimates are logical based on the wide range of diversity and pathogenicity of AMDV isolates. The results suggest that increases in the number of nucleotide substitutions over time and subsequent divergence make it difficult to accurately trace back AMDV isolates to their origin when several years elapsed between the two samplings.

Keywords: Aleutian mink disease virus, American mink, mutation rate, nucleotide substitution

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Authors: Dolores Hidalgo, Jesús M. Martín-Marroquín, Francisco Corona

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Municipal waste consist of a variety of items that are everyday discarded by the population. They are usually collected by municipalities and include waste generated by households, commercial activities (local shops) and public buildings. The composition of municipal waste varies greatly from place to place, being mostly related to levels and patterns of consumption, rates of urbanization, lifestyles, and local or national waste management practices. Each year, a huge amount of resources is consumed in the EU, and according to that, also a huge amount of waste is produced. The environmental problems derived from the management and processing of these waste streams are well known, and include impacts on land, water and air. The situation in remote areas is even worst. Difficult access when climatic conditions are adverse, remoteness of centralized municipal treatment systems or dispersion of the population, are all factors that make remote areas a real municipal waste treatment challenge. Furthermore, the scope of the problem increases significantly because the total lack of awareness of the existing risks in this area together with the poor implementation of advanced culture on waste minimization and recycling responsibly. The aim of this work is to analyze the existing situation in remote areas in reference to the production of municipal waste and evaluate the efficiency of different management alternatives. Ideas for improving waste management in remote areas include, for example: the implementation of self-management systems for the organic fraction; establish door-to-door collection models; promote small-scale treatment facilities or adjust the rates of waste generation thereof.

Keywords: door to door collection, islands, isolated areas, municipal waste, remote areas, rural communities

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Authors: Abderrezzak khatib

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Biogenic amines are low molecular weight nitrogenous compounds that have the potential to accumulate in food, posing a significant risk to food safety and human health. In this study, we investigated the inhibitory activity of three strains of lactic acid bacteria (LAB), against the growth and production of biogenic amines by both foodborne pathogens and food spoilage bacteria. The foodborne pathogens studied included Staphylococcus aureus, Pseudomonas aeruginosa, and Salmonella Paratyphi, while the food spoilage bacteria comprised Enterobacter cloacae and Proteus mirabilis. The methodology involved bacterial growth determination in petri dishes, bacterial culture extraction and derivatization, and biogenic amine analysis using HPLC. Our findings revealed that the inhibitory effects of LAB on these pathogens varied, with all three LAB strains demonstrating a remarkable reduction in the total bacterial count when combined with most pathogens, compared to the individual cultures of the pathogens. Furthermore, the presence of LAB in co-cultures with the pathogens resulted in a significant decrease in the production of tyramine and other biogenic amines by the pathogens themselves. These results suggest that LAB strains hold considerable promise in preventing the accumulation of biogenic amines in food products, thereby enhancing food safety. This study provides insights into the potential utilization of LAB in the context of preserving and ensuring the safety of food products. It highlights the significance of conducting additional research endeavors to elucidate the underlying mechanisms involved and to identify the precise bioactive compounds that are responsible for the observed inhibitory effects.

Keywords: food safety, lactic acid bacteria, foodborne pathogens, food spoilage bacteria, biogenic amines, tyrosine

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Authors: Tomasz Borowski, Dawid Sołoducha, Agata Markowska-Szczupak, Aneta Wesołowska, Marian Kordas, Rafał Rakoczy

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Essential oils (EOs) are fragrant volatile oils obtained from plants. These are used for cooking (for flavor and aroma), cleaning, beauty (e.g., rosemary essential oil is used to promote hair growth), health (e.g. thyme essential oil cures arthritis, normalizes blood pressure, reduces stress on the heart, cures chest infection and cough) and in the food industry as preservatives and antioxidants. Rosemary and thyme essential oils are considered the most eminent herbs based on their history and medicinal properties. They possess a wide range of activity against different types of bacteria and fungi compared with the other oils in both in vitro and in vivo studies. However, traditional uses of EOs are limited due to rosemary and thyme oils in high concentrations can be toxic. In light of the accessible data, the following hypothesis was put forward: Low frequency rotating magnetic field (RMF) increases the antimicrobial potential of EOs. The aim of this work was to investigate the antimicrobial activity of commercial Salvia Rosmarinus L. and Thymus vulgaris L. essential oil from Polish company Avicenna-Oil under Rotating Magnetic Field (RMF) at f = 25 Hz. The self-constructed reactor (MAP) was applied for this study. The chemical composition of oils was determined by gas chromatography coupled with mass spectrometry (GC-MS). Model bacteria Escherichia coli K12 (ATCC 25922) was used. Minimum inhibitory concentrations (MIC) against E. coli were determined for the essential oils. Tested oils in very small concentrations were prepared (from 1 to 3 drops of essential oils per 3 mL working suspensions). From the results of disc diffusion assay and MIC tests, it can be concluded that thyme oil had the highest antibacterial activity against E. coli. Moreover, the study indicates the exposition to the RMF, as compared to the unexposed controls causing an increase in the efficacy of antibacterial properties of tested oils. The extended radiation exposure to RMF at the frequency f= 25 Hz beyond 160 minutes resulted in a significant increase in antibacterial potential against E. coli. Bacteria were killed within 40 minutes in thyme oil in lower tested concentration (1 drop of essential oils per 3 mL working suspension). Rapid decrease (>3 log) of bacteria number was observed with rosemary oil within 100 minutes (in concentration 3 drops of essential oils per 3 mL working suspension). Thus, a method for improving the antimicrobial performance of essential oil in low concentrations was developed. However, it still remains to be investigated how bacteria get killed by the EOs treated by an electromagnetic field. The possible mechanisms relies on alteration in the permeability of ionic channels in ionic channels in the bacterial cell walls that transport in the cells was proposed. For further studies, it is proposed to examine other types of essential oils and other antibiotic-resistant bacteria (ARB), which are causing a serious concern throughout the world.

Keywords: rotating magnetic field, rosemary, thyme, essential oils, Escherichia coli

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Authors: Eglė Žurauskaitė, Eglė Gudavičienė

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The goal of this paper is to reveal how polarization is constructed through the use of impoliteness strategies. In general, media helps to spread various ideas very fast, and it means that processes of polarization are best revealed in computer-mediated communication (CMC) contexts. For this reason, data for the research was collected from online texts about a current, very diverse topic in Lithuania - Lithuanian language policy and regulations, because this topic is causing a lot of tension in Lithuanian society. Computer-mediated communication allows users to edit their message before they send it. It means that addressees carefully select verbal expressions to convey their message. In other words, each impoliteness strategy and its verbal expression were created intentionally. Impoliteness strategies in this research are understood as various ways to reach a communicative goal: belittle the other. To reach the goal, the public opinions of various Lithuanian public figures (e. g., cultural people, politicians, officials) were collected from new portals in 2019–2023 and analyzed using both quantitative and qualitative approaches. First, problematic aspects of the language policy, for which public figures complain, were identified. Then instances when public figures take a defensive position were analyzed: how they express this position and what it reveals about Lithuanian culture. Findings of this research demonstrate how concepts of impoliteness theory can be applied in analyzing the process of polarization in Lithuanian society on issues related to the State language policy. Also, to reveal how polarization is constructed, these tasks were set: a) determine which impoliteness strategies are used throughout the process of creating polarization, b) analyze how they were expressed verbally (e. g., as an advice, offer, etc.).

Keywords: impoliteness, Lithuanian language policy, polarization, impoliteness strategies

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Authors: A. A. Adedini, B. A. Aina, P. U. Ogbo

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Child obesity is on the rise globally. According to the World Health Organization, the number of obese children would increase to 70 million by 2025 if no intervention is made. An increase in the prevalence of overweight and obesity amongst school children in Lagos State, Nigeria has been established but specific factors promoting its prevalence are unknown. This aim of this study is to identify the commonly expressed factor(s) responsible for the rise in prevalence of child overweight and obesity in Lagos, Nigeria. Five focus group discussions were conducted with different groups of stake-holders involved in child care, namely: parents, teachers and health workers. Participants were recruited using a purposive sampling method; a validated question guide was employed for the discussion sessions. The discussions were recorded, collated, analysed using Grounded theory to extract themes. Six themes emerged from the discussions as follows: Civilization and lifestyle imbalance resulting from busy work schedules of young mothers leading to adoption of westernized culture promoting preference for processed and fast food meals; insecurity and congestion of the state which discourages out-door activities; ignorance of the populace on the prevalence of child obesity in the state; inadequate educative and enlightenment programmes in schools and by the Nigerian government; myths on child care and body physique and societal perceptions of the children born into affluent homes. Some of the factors responsible for the rise in the prevalence of child obesity in Lagos, Nigeria have been identified. Preventive strategies to control the prevalence of obesity in children residing in Lagos state is considered for further studies.

Keywords: Childhood Obesity, factors, lagos state, stakeholders

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Authors: Marwa Abdulsalam, Osamu Suzuki, Wirawan Dony Dahana

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One of the remarkable and unique industries in Japan is the food samples industry which can be noticed in most of the restaurants located around Japan. However, the market is getting saturated, which has pushed Japanese food sample manufacturers to start exploring new international markets. Most of the markets they explored were in the East Asian region, such as China or Korea. In this research, we examine the feasibility and the potential adoption rate of food samples in the international market outside the East Asian region. The main focus of this study is on the Saudi Arabian market. Nonetheless, since Saudi Arabia is a big market, the study results could possibly be applied to the international market as well. The study has conducted a quantitative survey to test the potential of the food samples industry in Saudi Arabia especially in 4 major cities: Jeddah, Mecca, Riyadh, and Dammam. The survey also tests the willingness to purchase, the average price point that the consumer is willing to pay for food samples, and the factors that drive restaurant owners to adopt the food samples system. The study created a correlation analysis between different factors, such as the geographic factor and the size of the restaurant factor, to examine the effect of different aspects on the purchasing decision. The study has found that the Japanese food samples system is predicted to adapt successfully in the Saudi Arabian market and in the international market alike due to the high importance of the food culture and the existence of the communication challenges that the food samples can solve. Additionally, the market survey stated in this study indicated that 83% of the restaurants’ managers are willing to adopt this system in their restaurants.

Keywords: food samples, innovative marketing, international market, marketing method

Procedia PDF Downloads 89

Authors: Dharmpal Singh

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The objective of this proposed project is to relocation and resettlement of carnivores tribal communities who are currently residing in the protected forest land in all over the world just like resettlement project of the carnivores tribal families of Mongia who at past were residing in Ranthambhore Tiger Reserve (RTR) and had caused excess damage of endangered species of wildlife including Tigers. At present several tribal communities are residing in the another national parks and they not only consuming the wild animals but also involved in illegal trading of vital organs, skin and bones with National and international traders. Tribal are ideally suited for the job because they are highly skilled game trackers and due to having had a definite source of income over the years, they easily drawn in to the illegal wildlife trade and slaughter of wild animals. Their income is increasing but wild animals are on the brink of extinction. For the conservation of flora and fauna the rehabilitation process should be thought out according to the RTR project (which not only totally change the quality of life of mongia tribal community but also increased the conopy cover of forest and grass due to reduced the biotic pressure on protected land of forest in Rajasthan state) with appropriate understanding of the sociology of the people involved, their culture, education standard and the need of different skills to be acquired by them for sustenance such as agriculture, dairy, poultry, social forestry, job as forest guard and others eco-development programmes. Perhaps, the dimensions presented by me may generate discussion among the international wild life lovers and conservationists and remedies may be result oriented in the field of management of forest and conservation of wildlife on this planet.

Keywords: strategies, rehablety of tribals, conservation of forest, eco-development Programmes, wildlife

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Authors: Richard Briginshaw, Hana Alaojeli, Javaria Ahmad, Hamza Gaffar, Nourtan Murad

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Sustainable renovations to cultural buildings and sites require a high level of competency in the sometimes conflicting areas of social/historical demands, environmental concerns, and the programmatic and technical requirements of the project. A detailed analysis of the existing site, building and client program are critical to reveal both challenges and opportunities. This forms the starting point for the design process – empirical explorations that search for a balanced and inspired architectural solution to the project. The Red Bay National Historic Site on the Labrador Coast of eastern Canada is a challenging project to explore and resolve these ideas. Originally the site of a 16ᵗʰ century whaling station occupied by Basque sailors from France and Spain, visitors now experience this history at the interpretive center, along with the unique geography, climate, local culture and vernacular architecture of the area. Working with our client, Parks Canada, the project called for significant alterations and expansion to the existing facility due to an increase in the number of annual visitors. Sustainable aspects of the design are focused on sensitive site development, passive energy strategies such as building orientation and building envelope efficiency, active renewable energy systems, carefully considered material selections, water efficiency, and interiors that respond to human comfort and a unique visitor experience.

Keywords: sustainability, renovations and expansion, cultural project, architectural design, green building

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Authors: María D. Groppa, Andrea Trentini, Myriam Zawoznik, Roxana Bigi, Carlos Nadra, Patricia L. Marconi

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In the present work, a remediation bioprocess based on the use of a local isolate of the microalgae Chlorella vulgaris immobilized in alginate beads is proposed. This process was shown to be effective for the reduction of several chemical and microbial contaminants present in Cildáñez stream, a water course that is part of the Matanza-Riachuelo Basin (Buenos Aires, Argentina). The bioprocess, involving the culture of the microalga in autotrophic conditions in a stirred-tank bioreactor supplied with a marine propeller for 6 days, allowed a significant reduction of Escherichia coli and total coliform numbers (over 95%), as well as of ammoniacal nitrogen (96%), nitrates (86%), nitrites (98%), and total phosphorus (53%) contents. Pb content was also significantly diminished after the bioprocess (95%). Standardized cytotoxicity tests using Allium cepa seeds and Cildáñez water pre- and post-remediation were also performed. Germination rate and mitotic index of onion seeds imbibed in Cildáñez water subjected to the bioprocess was similar to that observed in seeds imbibed in distilled water and significantly superior to that registered when untreated Cildáñez water was used for imbibition. Our results demonstrate the potential of this simple and cost-effective technology to remove urban-water contaminants, offering as an additional advantage the possibility of an easy biomass recovery, which may become a source of alternative energy.

Keywords: bioreactor, bioremediation, Chlorella vulgaris, Matanza-Riachuelo Basin, microalgae

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Authors: Eun-Ji Lee, Hye-Sung Park, Chan-Jung Lee, Won-Sik Kong

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We analyzed the DNA sequence of the ITS (Internal Transcribed Spacer) region of the 18S ribosomal gene and compared it with the gene sequence of T. fuciformis and Hypoxylon sp. in the BLAST database. The sequences of collected T. fuciformis and Hypoxylon sp. have over 99% homology in the T. fuciformis and Hypoxylon sp. sequence BLAST database. In order to select the optimal medium for T. fuciformis, five kinds of a medium such as Potato Dextrose Agar (PDA), Mushroom Complete Medium (MCM), Malt Extract Agar (MEA), Yeast extract (YM), and Compost Extract Dextrose Agar (CDA) were used. T. fuciformis showed the best growth on PDA medium, and Hypoxylon sp. showed the best growth on MCM. So as to investigate the optimum pH and temperature, the pH range was set to pH4 to pH8 and the temperature range was set to 15℃ to 35℃ (5℃ degree intervals). Optimum culture conditions for the T. fuciformis growth were pH5 at 25℃. Hypoxylon sp. were pH6 at 25°C. In order to confirm the most suitable carbon source, we used fructose, galactose, saccharose, soluble starch, inositol, glycerol, xylose, dextrose, lactose, dextrin, Na-CMC, adonitol. Mannitol, mannose, maltose, raffinose, cellobiose, ethanol, salicine, glucose, arabinose. In the optimum carbon source, T. fuciformis is xylose and Hypoxylon sp. is arabinose. Using the column test, we confirmed sawdust a suitable for T. fuciformis, since the composition of sawdust affects the growth of fruiting bodies of T. fuciformis. The sawdust we used is oak tree, pine tree, poplar, birch, cottonseed meal, cottonseed hull. In artificial cultivation of T. fuciformis with sawdust medium, T. fuciformis and Hypoxylon sp. showed fast mycelial growth on mixture of oak tree sawdust, cottonseed hull, and wheat bran.

Keywords: cultivation, optimal condition, tremella fuciformis, nutritional source

Procedia PDF Downloads 185

Authors: Malkhaz Jokhadze, Vakhtang Mshvildadze, Levan Makaradze, Ekaterine Mosidze, Salome Barbaqadze, Mariam Murtazashvili, Dali Berashvili, Koba sivsivadze, Lasha Bakuridze, Aliosha Bakuridze

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Essential oils are one of the most important groups of biologically active substances present in plants. Due to the chemical diversity of components, essential oils and their preparations have a wide spectrum of pharmacological action. They have bactericidal, antiviral, fungicidal, antiprotozoal, anti-inflammatory, spasmolytic, sedative and other activities. They are expectorant, spasmolytic, sedative, hypotensive, secretion enhancing, antioxidant remedies. Based on preliminary pharmacological studies, we have developed a formulation called “Phytobiotic” containing essential oils, a feed additive for poultry as an alternative to antibiotics. Phytobiotic is a water-soluble powder containing a composition of essential oils of thyme, clary, monarda and auxiliary substances: dry extract of liquorice and inhalation lactose. On this stage of research, the goal was to study the chemical composition of provided phytobiotic, identify the main substances and determine their quantity, investigate the biological activity of phytobiotic through in vitro and in vivo studies. Using gas chromatography-mass spectrometry, 38 components were identified in phytobiotic, representing acyclic-, monocyclic-, bicyclic-, and sesquiterpenes. Together with identification of main active substances, their quantitative content was determined, including acyclic terpene alcohol β-linalool, acyclic terpene ketone linalyl acetate, monocyclic terpenes: D-limonene and γ-terpinene, monocyclic aromatic terpene thymol. Provided phytobiotic has pronounced and at the same time broad spectrum of antibacterial activity. In the cell model, phytobiotic showed weak antioxidant activity, and it was stronger in the ORAC (chemical model) tests. Meanwhile anti-inflammatory activity was also observed. When fowls were supplied feed enriched with phytobiotic, it was observed that gained weight of the chickens in the experimental group exceeded the same data for the control group during the entire period of the experiment. The survival rate of broilers in the experimental group during the growth period was 98% compared to -94% in the control group. As a result of conducted researches probable four different mechanisms which are important for the action of phytobiotics were identified: sensory, metabolic, antioxidant and antibacterial action. General toxic, possible local irritant and allergenic effects of phytobiotic were also investigated. Performed assays proved that formulation is safe.

Keywords: clary, essential oils, monarda, poultry, phytobiotics, thyme

Procedia PDF Downloads 153

Authors: Tek Nath Subedi

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Violent activities used previously to erupt in specific societies owing to unequal, oppressive and unjust social practices have crossed out of their usual place of occurrence and entered the peace-zones such as schools and universities. The recent history of Universities in Nepal is marked with violent activities of students targeting to professors and other university officials. A student in a university is supposed to involve in scholarly activities and cooperate the teacher so as to achieve the academic degree and to enhance knowledge. Besides, the relationship between a teacher and a student is supposed to be the most sacred, which, in other words, is explained as the relationship between deity and devotee in a spiritual ground, carer and the kid in a moral ground, and mutually dependent in a functionalist view. Above all, the interdependence between teacher and student is illustrated as the two sides of the same coin. But the targeted attack on teachers and the degrading relationship between university and students raises serious questions on what makes a student violent and why the academia is terrorized. Therefore, this research aims to find the nexus of deviant activities of students to other social, political, economic and cultural areas of their life. To find possible answer of a few analytical questions, this paper takes an in-depth interview into consideration based on a snowball sampling and inquires some professors and officials how and why they are terrorized. It also reaches to some students who have witnessed or been part of any terror, following the same research method and ask them why violence has become recurring in the university and how university can be converted into a peace-zone. This research has simply hypothesized that the terror-inspired activities carried out by students in the university are the extended version of their deviant activities out of the university premises. But, it was found that the deviant activities of students were connected in particular to their culture and political identity.

Keywords: deviance, relationship, student, teacher, terror, university

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Authors: Soyhan Egitim

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This case study aims to investigate leadership practices in diversity management in the liberal arts department of a Japanese university. In 2013, the Japanese Ministry of Education, Sports, Science, and Technology (MEXT) revealed their English education reform plan in response to rapid globalization. Based on the new reform plan, Japanese universities would expand their international faculty in order to promote globalization through an increased number of intercultural communication and content-based language classes in English. The study employed a two-phased qualitative approach to gain a deeper understanding of the management strategies employed in diversity management, and the leadership practices influenced those management strategies. In the first phase, a closed-ended qualitative survey was conducted with ten adjunct faculty members from the liberal arts department. The results indicate that syllabus design, grading scheme, textbook choices, and class management policies are strictly regulated by the tenured Japanese faculty. In the second phase, semi-structured interviews were held with international faculty members to understand their personal experiences. Their responses revealed that top-down management approaches are counter-effective in the department’s efforts to promote diversity and thus, a new organizational culture needs to be nurtured to emphasize inclusion alongside diversity. In this regard, the study proposes collaborative leadership as an inclusive leadership practice to minimize power differences in the hierarchy and increase opportunities for inclusion in the rapidly diversifying workforce.

Keywords: collaborative leadership, diversity, inclusion, international faculty, top-down

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Authors: M. Y. Abubakar, M. Yunisa, A. N. Muhammad

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Constraining the production Clarias gariepinus and Oreochromis niloticus culture is the prohibitive cost of feed. We assess the performance of the species fingerlings on diets substituted with composite. Four dietary treatments (0%, 25%, 45%, and 75%) for C. gariepinus and five (0%, 25%, 50%, 75%, and whole food composite) for O. niloticus were formulated and each fed to 15 fingerlings for C. gariepinus and 10 fingerlings for O. niloticus stocked in 75ltrs plastic bowls, replicated trice in a completely randomized design. The experiment lasted 56 days. Percent survival rate was significantly (p < 0.05) higher (57.78 ± 9.69) in C. gariepinus fed diet III. The growth and nutrient utilization indices were least in the fish fed diet IV, which was significantly (p < 0.05) lower than in other treatments. Fish fed dietary treatment III, recorded the best in growth and nutrient utilization indices and was significantly higher (p < 0.05) than those fed dietary treatments I & II which were non-significant (p > 0.05) and higher than those fed 75% substitution. Better profit index was in the fish fed diet with 50% substitution level. For O. niloticus, the survival (172.62 ± 39.03) was significantly higher (p < 0.05) in those fed 25% substituted diet. For growth indices, the least performed were those fed whole composite while other treatments were non-significant (p > 0.05) different from each other. In terms of nutrient utilization, fish fed diet substituted at 0%, 25%, 50% and 75% food composite had similar food conversion ratio and protein efficiency ratio. However, there was no significant difference in the profit index among the whole treatment. It can be concluded that food composite from Sokoto house-holds can optimally replace groundnut cake up to 50% level as a protein source in the diets of Clarias gariepinus and O. niloticus fingerlings without adverse effects on survival, growth, and nutrient utilization.

Keywords: food composite, nutrient utilization, C. gariepinus, O. niloticus household, substitution levels

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Authors: Yetti Marlida, Harnentis, Yuliaty Shafan Nur

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Background: Tempoyak is a dish derived from fermented durian fruit. Tempoyak is a food consumed as a side dish when eating rice. Besides being eaten with rice, tempoyak can also be eaten directly. But this is rarely done because many cannot stand the sour taste and aroma of the tempoyak itself. In addition, tempoyak can also be used as a seasoning. The taste of tempoyak is acidic, this occurs because of the fermentation process in durian fruit meat which is the raw material. Tempoyak is already very well known in Indonesia, especially in Padang, Bengkulu, Palembang, Lampung, and Kalimantan. Besides that, this food is also famous in Malaysia. The purpose of this research is to improvement production of γ-aminobutyric acid (GABA) by Lactobacillus plantarum isolated from indigenous fermented durian (tempoyak). Selected Lactic Acid Bacteria (LAB) previously isolated from indigenous fermented durian (tempoyak) that have ability to produce γ-aminobutyric acid (GABA). The study was started with identification of selected LAB by 16 S RNA, followed optimation of GABA production by culture condition using different initial pH, temperature, glutamate concentration, incubation time, carbon and nitrogen sources. Results: The result from indentification used polymerase chain reaction of 16S rRNA gene sequences and phylogenetic analysis was Lactobacillus plantarum (coded as Y3) with a sequenced length of 1400bp. The improvement of Gaba production was found highest at pH: 6.0; temperature: 30 °C; glutamate concentration: 0.4%; incubation time: 60 h; glucose and yeast extract as carbon and nitrogen sources. Conclusions: GABA can be produced with the optimum condition fermentation were 66.06 mM.

Keywords: lactic acid bacteria, γ-amino butyric acid, indigenous fermented durian, PCR

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Authors: Lemar Blake, Chris Oura, Ayanna C. N. Phillips Savage

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Improved DNA sequencing technology has greatly enhanced bacterial identification, especially for organisms that are difficult to culture. Mycobacteriosis with consistent hyphema, bilateral exophthalmia, open mouth gape and ocular lesions, were observed in various fish populations at the School of Veterinary Medicine, Aquaculture/Aquatic Animal Health Unit. Objective: To identify the species of Mycobacterium that is affecting aquarium fish at the School of Veterinary Medicine, Aquaculture/Aquatic Animal Health Unit. Method: A total of 13 fish samples were collected and analyzed via: Ziehl-Neelsen, conventional polymerase chain reaction (PCR) and real-time PCR. These tests were carried out simultaneously for confirmation. The following combination of conventional primers: 16s rRNA (564 bp), rpoB (396 bp), sod (408 bp) were used. Concatenation of the gene fragments was carried out to phylogenetically classify the organism. Results: Acid fast non-branching bacilli were detected in all samples from homogenized internal organs. All 13 acid fast samples were positive for Mycobacterium via real-time PCR. Partial gene sequences using all three primer sets were obtained from two samples and demonstrated a novel strain. A strain 99% related to Mycobacterium marinum was also confirmed in one sample, using 16srRNA and rpoB genes. The two novel strains were clustered with the rapid growers and strains that are known to affect humans. Conclusions: Phylogenetic analysis demonstrated two novel Mycobacterium strains with the potential of being zoonotic and one strain 99% related to Mycobacterium marinum.

Keywords: polymerase chain reaction, phylogenetic, DNA sequencing, zoonotic

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Authors: Arieh Moussaieff, Bénédicte Elena-Herrmann, Yaakov Nahmias, Daniel Aberdam

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Differentiation of pluripotent stem cells is a slow process, marked by the gradual loss of pluripotency factors over days in culture. While the first few days of differentiation show minor changes in the cellular transcriptome, intracellular signaling pathways remain largely unknown. Recently, several groups demonstrated that the metabolism of pluripotent mouse and human cells is different from that of somatic cells, showing a marked increase in glycolysis previously identified in cancer as the Warburg effect. Here, we sought to identify the earliest metabolic changes induced at the first hours of differentiation. High-resolution NMR analysis identified 35 metabolites and a distinct, gradual transition in metabolism during early differentiation. Metabolic and transcriptional analyses showed the induction of glycolysis toward acetate and acetyl-coA in pluripotent cells, and an increase in cholesterol biosynthesis during early differentiation. Importantly, this metabolic pathway regulated differentiation of human and mouse embryonic stem cells. Acetate delayed differentiation preventing differentiation-induced histone de-acetylation in a dose-dependent manner. Glycolytic inhibitors upstream of acetate caused differentiation of pluripotent cells, while those downstream delayed differentiation. Our data suggests that a rapid loss of glycolysis in early differentiation down-regulates acetate and acetyl-coA production, causing a loss of histone acetylation and concomitant loss of pluripotency. It demonstrate that pluripotent stem cells utilize a novel metabolism pathway to maintain pluripotency through acetate/acetyl-coA and highlights the important role metabolism plays in pluripotency and early differentiation of stem cells.

Keywords: pluripotency, metabolomics, epigenetics, acetyl-coA

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