Search results for: exosomes isolation on SPE

Authors: Ainur Mukhambetova, Miras Karzhauov, Vyacheslav Ogay

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Introduction: Mesenchymal stem cells (MSCs) have the capacity to be differentiated into several cell lineages and are a promising source for cell therapy and tissue engineering. However, currently most MSCs culturing protocols use media supplemented with fetal bovine serum (FBS), which limits their application in clinic due to the possibility of zoonotic infections, contamination and immunological reactions. Consequently, formulating effective serum free culture medium becomes one of the important problems in contemporary cell biotechnology. Objectives: The aim of this study was to define an optimal serum-free medium for culturing of periosteum derived MSCs. Materials and methods: The MSCs were extracted from human periosteum and transferred to the culture flasks pretreated with CELLstart™. Immunophenotypic characterization, proliferation and in vitro differentiation of cells grown on STEM PRO® MSC SFM were compared to the cells cultured in the standard FBS containing media. Chromosome analysis and flow cytometry were also performed. Results: We have shown that cells were grown on STEM PRO® MSC SFM retained all the morphological, immunophenotypic (CD73, CD90, CD105, vimentin and Stro-1) and cell differentiation characteristics specific to MSCs. Chromosome analysis indicated no anomalies in the chromosome structure. Flow cytometry showed a high expression of cell adhesion molecules CD44 (98,8%), CD90 (97,4%), CD105 (99,1%). In addition, we have shown that cell is grown on STEM PRO® MSC SFM have higher proliferation capacity compared to cell expanded on standard FBS containing the medium. Conclusion: We have shown that STEM PRO® MSC SFM is optimal for culturing periosteum derived human MSCs which subsequently can be safely used in cell therapy.

Keywords: cell technologies, periosteum-derived MSCs, regenerative medicine, serum-free medium

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Authors: Rukshika S Hewawasam, Sisira K. Weliwegamage, Sanath Rajapakse, Subramanium Sotheeswaran

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Bio fuel is one of the emerging industries around the world due to arise of crisis in petroleum fuel. Fermentation is a cost effective and eco-friendly process in production of bio-fuel. So inventions in microbes, substrates, technologies in fermentation cause new modifications in fermentation. One major problem in microbial ethanol fermentation is the low resistance of conventional microorganisms to the high ethanol concentrations, which ultimately lead to decrease in the efficiency of the process. In the present investigation, an ethanol resistant bacterium was isolated from sap of Saccharum officinarum (sugar cane). The optimal cultural conditions such as pH, temperature, incubation period, and microbiological characteristics, morphological characteristics, biochemical characteristics, ethanol tolerance, sugar tolerance, growth curve assay were investigated. Isolated microorganism was tolerated to 18% (V/V) of ethanol concentration in the medium and 40% (V/V) glucose concentration in the medium. Biochemical characteristics have revealed as Gram negative, non-motile, negative for Indole test ,Methyl Red test, Voges- Proskauer`s test, Citrate Utilization test, and Urease test. Positive results for Oxidase test was shown by isolated bacterium. Sucrose, Glucose, Fructose, Maltose, Dextrose, Arabinose, Raffinose, Lactose, and Sachcharose can be utilized by this particular bacterium. It is a significant feature in effective fermentation. The fermentation process was carried out in glucose medium under optimum conditions; pH 4, temperature 30˚C, and incubated for 72 hours. Maximum ethanol production was recorded as 12.0±0.6% (V/V). Methanol was not detected in the final product of the fermentation process. This bacterium is especially useful in bio-fuel production due to high ethanol tolerance of this microorganism; it can be used to enhance the fermentation process over conventional microorganisms. Investigations are currently conducted on establishing the identity of the bacterium

Keywords: bacterium, bio-fuel, ethanol tolerance, fermentation

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Authors: Asteria Nsamba

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Social networking applications and tools have become compelling platforms for generating and sharing knowledge across the world. Social networking applications and tools refer to a variety of social media platforms which include Facebook, Twitter WhatsApp, blogs and Wikis. The most popular of these platforms are Facebook, with 2.41 billion active users on a monthly basis, followed by WhatsApp with 1.6 billion users and Twitter with 330 million users. These communication platforms have not only impacted social lives but have also impacted students’ learning, across different delivery modes in higher education: distance, conventional and blended learning modes. With this amount of interest in these platforms, knowledge sharing has gained importance within the context in which it is required. In open distance learning (ODL) contexts, social networking platforms can offer students and teachers the platform on which to create and share knowledge, and form learning collaborations. Thus, they can serve as support mechanisms to increase interactions and reduce isolation and loneliness inherent in ODL. Despite this potential and opportunity, research indicates that many ODL teachers are not inclined to using social media tools in learning. Although it is unclear why these tools are uncommon in these environments, concerns raised in the literature have indicated that many teachers have not mastered the art of teaching with technology. Using technological, pedagogical content knowledge (TPCK) and product quality theory, and Bloom’s Taxonomy as lenses, this paper is aimed at; firstly, assessing the quality of three social media applications: Facebook, Twitter and WhatsApp, in order to determine the extent to which they are suitable platforms for teaching and learning, in terms of content generation, information sharing and learning collaborations. Secondly, the paper demonstrates the application of teaching, learning and assessment using Bloom’s Taxonomy.

Keywords: distance education, quality, social networking tools, TPACK

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Authors: Muhammad Sohail Sajid, Mahvish Maqbool, Hafiz Muhammad Rizwan, Muhammad Saqib, Haroon Ahmad

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Ticks are blood-sucking ectoparasite that causes a decrease in production and economic losses and affects mammals, reptiles, and birds. Hyalomma anatolicum is the main vector for CCHF transmission and Pakistan has faced several outbreaks of CCHF in the recent past. Ferritin (fer)is a highly conserved molecule that is ubiquitous in most tick tissues and responsible for iron metabolism and storage. It was hypothesized that the development of acaricidal resistance and residual effects of commercially used acaricides could be controlled by using alternative control methods, including RNA interference. The current study aimed to evaluate the fer silencing effects on tick feeding, average body weight, egg mass index, and mortality. Ticks, collected through the standard collection protocols were further subjected to RNA isolation using the Trizol method. Commercially available kit procedures were followed for cDNA and dsRNA synthesis. The soaking/Immersion method was used for dsRNA delivery. Our findings have shown a 27% reduction in body weight of fer silenced group and showed a significant association of fer and body weight. Silencing of fer had a significant effect on the engorgement percentage (P= 0.0007), oviposition (P=0.008), egg mass (P= 0.004) and hatching (P= 0.001). The soaking method was used for dsRNA delivery and 15°C was found to be an optimum temperature for inducing gene silencing in ticks as at this temperature, maximum survivability after immersion was attained. This study along with previous studies, described that iron toxicity due to the silencing of fer could play an important role in the control of ticks and fer can be used as a potent candidate for vaccine development.

Keywords: ticks, iron, ferritin, engorgement, oviposition, immersion, RNA interference

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Authors: Mohammad Aladwan, Adelia Skripova

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Bioremediation aspects of crude oil-polluted fields can be achieved by isolation and identification of bacterial species from oil-contaminated soil in order to choose the most active isolates and increase the strength of others. In this study, oil-degrading bacteria were isolated and identified from oil-contaminated soil samples in northeastern Jordan. The bacterial growth count (CFU/g) was between 1.06×10⁵ and 0.75×10⁹. Eighty-two bacterial isolates were characterized by their morphology and biochemical tests. The identified bacterial genera included: Klebsiella, Staphylococcus, Citrobacter, Lactobacillus, Alcaligenes, Pseudomonas, Hafnia, Micrococcus, Rhodococcus, Serratia, Enterobacter, Bacillus, Salmonella, Mycobacterium, Corynebacterium, and Acetobacter. Molecular identification of a universal primer 16S rDNA gene was used to identify four bacterial isolates: Microbacterium esteraromaticum strain L20, Pseudomonas stutzeri strain 13636M, Klebsilla pneumoniae, and uncultured Klebsilla sp., known as new strains. Our results indicate that their specific oil-degrading bacteria isolates might have a high strength of oil degradation from oil-contaminated sites. Staphylococcus intermedius (75%), Corynebacterium xerosis (75%), and Pseudomonas fluorescens (50%) showed a high growth rate on different types of hydrocarbons, such as crude oil, toluene, naphthalene, and hexane. In addition, monooxygenase and catechol 2,3-dioxygenase were detected in 17 bacterial isolates, indicating their superior hydrocarbon degradation potential. Total petroleum hydrocarbons were analyzed using gas chromatography for soil samples. Soil samples M5, M7, and M8 showed the highest levels (43,645, 47,805, and 45,991 ppm, respectively), and M4 had the lowest level (7,514 ppm). All soil samples were analyzed for heavy metal contamination (Cu, Cd, Mn, Zn, and Pb). Site M7 contains the highest levels of Cu, Mn, and Pb, while Site M8 contains the highest levels of Mn and Zn. In the future, these isolates of bacteria can be used for the cleanup of oil-contaminated soil.

Keywords: bioremediation, 16S rDNA gene, oil-degrading bacteria, hydrocarbons

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Authors: A. G. Barua, Uttam Rajkhowa, Pranjal Moni Nath, Nur Abdul Kadir

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Mycobacterium tuberculosis (MTB) is one of the most closely-related intracellular bacterial pathogens, grouped as the M. tuberculosis complex (MTC). MTB, the primary agent of human tuberculosis (TB), can develop clinical TB in animals as 75 percent of canine mycobacterial infection is caused by close contact with an infected human being. In the present study, molecular detection of TB in dogs in three North-eastern states of India, Assam Mizoram, and Nagaland was carried out. So far, there has been a lack of systematic study in these regions, hampered by slow diagnostic methods and poor infrastructure. In an attempt to rectify this situation, molecular epidemiology was carried out for nine months to detect canine TB in a sample of 340 dogs. Isolation of DNA was done with swabs (throat/nasal), nodules of lungs and fluids from 100 suspected dogs and the molecular study were carried out with the help of conventional and real-time PCR. Post-mortem study was also carried out. Our results showed that the prevalence of clinical TB in dogs from a high-risk setting was 1 percent. However, the prevalence of immunological sensitization to M. tuberculosis antigen in dogs living in contact with sputum smeared positive TB cases was almost 50 percent. The latter setting had the maximum impact in terms of TB transmission. During the study period, a survey with a standard questionnaire was carried out in the TB hospitals to study reverse zoonosis. It was observed that an infected human being was one of the major risk factors for dogs to contract the infection. This observation was drawn by examining the probable airborne transmission from humans to their pets or strays. The present study helped to discover the nuances of TB transmission more clearly and systematically as compared to other sporadic tests to detect MTB in canine.

Keywords: Assam and Nagaland, canine TB, India, molecular detection, tuberculosis

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Authors: Suvalux Chaichuchote, Ratchadaporn Thonghem

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Pleurotus mushroom is one of popular edible mushrooms in Thailand. It is much favored by consumers due to its delicious taste and high nutrition. It is commonly used as an ingredient in several dishes. The commercially cultivated strain grown in most farms is the Pleurotus sp., Hed Bhutan, that is widely distributed to mushroom farms throughout the country and can be cultivated almost all year round. However, it demands different cultivated strains from mushroom growers, therefore, the improving mushroom strains should be done to their benefits. In this study, we used a di-mon mating method to hybrid production from Hed Bhutan (P-3) as dikaryon material and monokaryotic mycelium were isolated from basidiospores of other three Pleurotus sp. by single spore isolation. The 3 hybrids: P-3XSA-6, P-3XSB-24 and P-3XSE-5 were recognized from the 12 hybridized successfully. They were appropriate hybridized in terms of fruiting body performance in the three time cycles of cultivation such as the number of days until growing, time for pinning, color and shape of fruiting bodies and yield. For genetic study, genomic DNAs of both Hed Bhutan (P-3) and three hybrids were extracted. A couple of primer ITS1 and ITS4 were used to amplify the gene coding for ITS1, ITS2 and 5.8S rRNA. The similarities between these amplified genes and databases of DNA revealed that Hed Bhutan (P-3) was the Pleurotus pulmonarius as well as P-3XSA-6, P-3XSB-24 and P-3XSE-5 hybrids. Furthermore, Hed Bhutan (P3) and three hybrids were distributed to 3 small-scale farms, with mushroom farming experience, in the countryside. To address this, one hundred and twenty mushroom bags of each strain were supplied to them. The findings, by interview, indicated two mushroom farmers were satisfied with P-3XSA-6 hybrid and P-3XSB-24 hybrid, thanks to their simultaneous fruiting time and good yield. While the other was satisfied with P-3XSB-24 hybrid due to its good yield and P-3XSE-5 hybrids thanks to its gradually fruiting body, benefiting in frequent harvest. Overall, farmers adopted all hybrids to grow as commercially cultivated strains as well as Hed Bhutan (P-3) strain.

Keywords: dikaryon, monokaryon, pleurotus, strain improvement

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Authors: Meenakshi Chaudhary, V .S. Randhawa, M. Jais, R. Dutta

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Introduction: An outbreak of Multi drug resistant S. Typhi (i.e. resistance to chloramphenicol, ampicillin, and trimethoprim-sulfamethoxazole) occurred in 1990's in India which peaked in 1992-93 and resulted in the change of drug of choice from chloramphenicol to ciprofloxacin for enteric fever. Currently an emergence of Ciprofloxacin susceptible S. Typhi isolates in the region is being reported which appears to be chromosomally mediated. Methodology: Six hundred sixty four strains were randomly selected from the time period between January 2008-December 2011 at the National Salmonella Phage Typing Centre, LHMC, New Delhi. The strains were representative of the north, central and south zones of India. All isolates were subjected to serotyping, biotyping, phage typing and then to antimicrobial susceptibility testing by CLSI disk diffusion (CLSI) technique to Ciprofloxacin, Cefotaxime, Ampicillin, Chloramphenicol, Trimethoprim-Sulfomethoxazole and Tetracycline. Subsequently MIC of the isolates was determined by E-test (AB-Biodisc). Results: More than 80% of the tested strains had intermediate susceptibility to ciprofloxacin. The E test revealed the MIC (Ciprofloxacin) of these strains to be in the range of 0.12 to 0.5 µg/ml. Sixty nine percent of ciprofloxacin intermediate susceptible strains belonged to Phage type E1 and fourteen percent of these were Vi- Negative i.e these could not be typed by the phage typing scheme of Craigie and Yen. All the strains remained susceptible to cefotaxime. Conclusion: Predominant isolation of intermediate susceptible S. Typhi strains from India would alter the recommendations of empiric treatment of enteric fever in the region. Alternative to the low cost ciprofloxacin will have to be sought or increased dosage and/or duration of ciprofloxacin will have to be recommended. The reasons for the trend of increase in percentage of intermediate susceptible S. Typhi strains are not clear but may be attributed partly to the revision of CLSI guidelines in 2013.

Keywords: salmonella typhi, decreased ciprofloxacin susceptibility, ciprofloxacin, minimum inhibitory concentration

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Authors: Wilfred Jeyatheese Jeyaraj

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Education is a key determinant of future success, and every person deserves non-discriminant access to information for educational inevitabilities in any case. Analysing and understanding complex information is a crucial learning tool, especially for students. In order to compete equally with sighted students, visually impaired students require the unhinged access to access to all the available information resources. When the education of visually impaired students comes to a focal point, it can be stated that visually impaired students encounter several obstacles and barriers before they enter the university and during their time there as students. These obstacles and barriers are spread across technical, organizational and social arenas. This study reveals the possible approaches to absorb and benefit from the information provided by the Sri Lankan University Libraries for visually impaired students. Purposive sampling technique was used to select sample visually impaired students attached to the Sri Lankan National universities. There are 07 National universities which accommodate the visually impaired students and with the identified data, they were selected for this study and 80 visually impaired students were selected as the sample group. Descriptive type survey method was used to collect data. Structured questionnaires, interviews and direct observation were used as research instruments. As far as the Sri Lankan context spread is concerned, visually impaired students are able to finish their courses through their own determination to overcome the barriers they encounter on their way to graduation, through moral and practical support from their own friends and very often through a high level of creativity. According to the findings there are no specially trained university librarians to serve visually impaired users and less number of assistive technology equipment are available at present. This paper enables all university libraries in Sri Lanka to be informed about the social isolation of visually compromised students at the Sri Lankan universities and focuses on the rectification issues by considering their distinct case for interaction.

Keywords: information access, Sri Lanka, university libraries, visual impairment

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Authors: Rashmi Tyagi, Savita Vermani

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Violence against women is the result of long-standing power imbalance between men and women and thus seriously compromises the well-being, productivity and contribution of one half the population. The costs incurred to the family especially children and society at large in terms of physical, psychological, social and financial losses are huge. The communities’ native to the state of Haryana in India is primarily patriarchal, burdened with age old regressive mindset under the socio-cultural and religious structures which discriminates against women. Therefore it was important to bring to light the issues affecting women in this region. Therefore this study focused on studying the consequences of crime on victim women and their families. Two hundred women were randomly selected and out of those one hundred twenty, who were affected with some kind of violence were interviewed. Data was collected and statistically analyzed for physical, psychological, inter-family and societal consequences of violence on these women. Women reported physical injuries, gynecological problems, unwanted pregnancies, frigidity, phobia and sexual dysfunction. 58.9% women felt decreased work efficiency. Psychological problems encountered were anxiety, isolation, depression, suicidal tendencies. 66.7% respondents suffered from anxiety followed by 65.0% faced depression symptoms. At family levels, 40.0% respondents felt the atmosphere was unsuitable for children while 39.2% reported lack of interaction. The societal consequences reported were breakdown of interaction with friends and family (44.2%) and resulting humiliation and demeaning remarks from others (38.3%). The impact of violence on women had an adverse effect on children. 36.7% children felt responsible for abuse and powerless to stop it, 29.2% reported living with fear. Concerted efforts are required to curb violence against women in Haryana.

Keywords: impact of violence against women on children, patriarchal society, physical psychological and societal consequences, violence against women

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Authors: Lutendo Nendauni

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Colonialism breaks things, colonisers exploded native cultural solidarity, producing the spiritual confusion, psychic wounding, and economic exploitation of a new and dominated ‘other’. Colonialism as the cultural and economic exploitation began when the West defended in their seizure of foreign territories for the exploitation of its natural resources; this resulted in brutal socio-economic imbalances. The Western profited at the detriment of the weak Africa. However, colonialism has since passed, but the effects are still evident culturally, socially, and economically. This paper explored how postcolonial writers and critics attempt to reverse the socio-economic imbalances resulting from the fragmentation of colonialism, with a focus on the play 'I will Marry When I Want' by Ngugi wa Thiong’o and Ngugi wa Mirii, as a primary text. Using qualitative discourse-textual analysis as the research methodology, the researcher purposively extracts discourse segments from the text for analysis and interpretation. The findings reveal that Postcolonial critics and writers attempt to reverse the socio-economic effects of colonialism through various counter discourses; their literature is concerned with the destruction of colonised identity, the search for this identity, and its assertion. It is manifest in the text that writers offer corrective views about Africans; they stress that they write their literary texts to conscientise their fellow Africans. Postcolonial writers and critics argue that language is a carrier of culture and that the only way to break free from colonial influence is by not adopting a foreign language. They further through their poems, novels, plays, and music strategically shine the spotlight on the previously nameless and destitute people so that they can develop the human spirit’s desire to overcome defeat, socio-political deprivation, and isolation.

Keywords: colonialism, postcoloniality, critics, socio-economic imbalances

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Authors: Sudhir Kumar Tewatia, Kanishck Tewatia, Anttriksh Tewatia

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Advancements in soil consolidation are discussed, and further improvements are proposed with particular reference to Tewatia’s Y-Y’ Approach, which is called the Settlement versus Rate of Settlement Approach in consolidation. A branch of calculus named Y-Y' (or y versus dy/dx) is suggested (as compared to the common X-Y', x versus dy/dx, dy/dx versus x or Newton-Leibniz branch) that solves some complicated/unsolved theoretical and practical problems in physical sciences (Physics, Chemistry, Mathematics, Biology, and allied sciences) and engineering in an amazingly simple and short manner, particularly when independent variable X is unknown and X-Y' Approach can’t be used. Complicated theoretical and practical problems in 1D, 2D, 3D Primary and Secondary consolidations with non-uniform gradual loading and irregularly shaped clays are solved with elementary school level Y-Y' Approach, and it is interesting to note that in X-Y' Approach, equations become more difficult while we move from one to three dimensions, but in Y-Y' Approach even 2D/3D equations are very simple to derive, solve, and use; rather easier sometimes. This branch of calculus will have a far-reaching impact on understanding and solving the problems in different fields of physical sciences and engineering that were hitherto unsolved or difficult to be solved by normal calculus/numerical/computer methods. Some particular cases from soil consolidation that basically creeps and diffusion equations in isolation and in combination with each other are taken for comparison with heat transfer. The Y-Y’ Approach can similarly be applied in wave equations and other fields wherever normal calculus works or fails. Soil mechanics uses mathematical analogies from other fields of physical sciences and engineering to solve theoretical and practical problems; for example, consolidation theory is a replica of the heat equation from thermodynamics with the addition of the effective stress principle. An attempt is made to give them mathematical analogies.

Keywords: calculus, clay, consolidation, creep, diffusion, heat, settlement

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Authors: Ali Aydin, Mert Sudagidan, Aysen Coban, Alparslan Kadir Devrim

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Pseudomonas spp. (specifically, P. fluorescens and P. fragi) are considered the principal spoilage microorganisms of refrigerated poultry meats. The higher the level psychrophilic spoilage Pseudomonas spp. on carcasses at the end of processing lead to decrease the shelf life of the refrigerated product. The aim of the study was the identification of psychrophilic Pseudomonas spp. having proteolytic and lipolytic activities from poultry meats by 16S rRNA and rpoB gene sequencing, investigation of protease and lipase related genes and determination of proteolytic activity of Pseudomonas spp. In the of isolation procedure, collected chicken meat samples from local markets and slaughterhouses were homogenized and the lysates were incubated on Standard method agar and Skim Milk agar for selection of proteolytic bacteria and tributyrin agar for selection of lipolytic bacteria at +4 °C for 7 days. After detection of proteolytic and lipolytic colonies, the isolates were firstly analyzed by biochemical tests such as Gram staining, catalase and oxidase tests. DNA gene sequencing analysis and comparison with GenBank revealed that 126 strong enzyme Pseudomonas spp. were identified as predominantly P. fluorescens (n=55), P. fragi (n=42), Pseudomonas spp. (n=24), P. cedrina (n=2), P. poae (n=1), P. koreensis (n=1), and P. gessardi (n=1). Additionally, protease related aprX gene was screened in the strains and it was detected in 69/126 strains, whereas, lipase related lipA gene was found in 9 Pseudomonas strains. Protease activity was determined using commercially available protease assay kit and 5 strains showed high protease activity. The results showed that psychrophilic Pseudomonas strains were present in chicken meat samples and they can produce important levels of proteases and lipases for food spoilage to decrease food quality and safety.

Keywords: Pseudomonas, chicken meat, protease, lipase

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Authors: Daniel O. Johnson, Abiodun A. Ogunseye, Aaron Aransiola, Majors Samuel

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Power Inverter is a very important device in renewable energy use particularly for solar photovoltaic power application because it is the effective interface between the DC power generator and the load or the grid. Transformerless inverter is getting more and more preferred to the power converter with galvanic isolation transformer and may eventually supplant it. Transformerless inverter offers advantages of improved DC to AC conversion and power delivery efficiency; and reduced system cost, weight and complexity. This work presents thorough analysis of the design and prototyping of 3KVA grid-tie transformerless inverter. The inverter employs electronic switching method with minimised heat generation in the system and operates based on the principle of pulse-width modulation (PWM). The design is such that it can take two inputs, one from PV arrays and the other from Battery Energy Storage BES and addresses the safety challenge of leakage current. The inverter system was designed around microcontroller system, modeled with Proteus® software for simulation and testing of the viability of the designed inverter circuit. The firmware governing the operation of the grid-tied inverter is written in C language and was developed using MicroC software by Mikroelectronica® for writing sine wave signal code for synchronization to the grid. The simulation results show that the designed inverter circuit performs excellently with very high efficiency, good quality sinusoidal output waveform, negligible harmonics and gives very stable performance under voltage variation from 36VDC to 60VDC input. The prototype confirmed the simulated results and was successfully synchronized with the utility supply. The comprehensive analyses of the circuit design, the prototype and explanation on overall performance will be presented.

Keywords: grid-tied inverter, leakage current, photovoltaic system, power electronic, transformerless inverter

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Authors: Tamrat Tesfaye, Bruce Sithole

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Box-Behnken Response surface methodology was used to determine the optimum processing conditions that give maximum extraction yield and whiteness index from mango seed. The steeping time ranges from 2 to 12 hours and slurring of the steeped seed in sodium metabisulphite solution (0.1 to 0.5 w/v) was carried out. Experiments were designed according to Box-Behnken Design with these three factors and a total of 15 runs experimental variables of were analyzed. At linear level, the concentration of sodium metabisulphite had significant positive influence on percentage yield and whiteness index at p<0.05. At quadratic level, sodium metabisulphite concentration and sodium metabisulphite concentration2 had a significant negative influence on starch yield; sodium metabisulphite concentration and steeping time*temperature had significant (p<0.05) positive influence on whiteness index. The adjusted R2 above 0.8 for starch yield (0.906465) and whiteness index (0.909268) showed a good fit of the model with the experimental data. The optimum sodium metabisulphite concentration, steeping hours, and temperature for starch isolation with maximum starch yield (66.428%) and whiteness index (85%) as set goals for optimization with the desirability of 0.91939 was 0.255w/v concentration, 2hrs and 50 °C respectively. The determined experimental value of each response based on optimal condition was statistically in accordance with predicted levels at p<0.05. The Mango seeds are the by-products obtained during mango processing and possess disposal problem if not handled properly. The substitution of food based sizing agents with mango seed starch can contribute as pertinent resource deployment for value-added product manufacturing and waste utilization which might play significance role of food security in Ethiopia.

Keywords: mango, synthetic sizing agent, starch, extraction, textile, sizing

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Authors: Kanika Arora, Madhu Bala

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The small intestine epithelium is highly sensitive and major targets of ionizing radiation. Radiation causes gastrointestinal toxicity either by direct deposition of energy or indirectly (inflammation or bystander effects) generating free radicals and reactive oxygen species. Oxidative stress generated as a result of radiation causes active inflammation within the intestinal mucosa leading to structural and functional impairment of gut epithelial barrier. As a result, there is a loss of tolerance to normal dietary antigens and commensal flora together with exaggerated response to pathogens. Dysbiosis may therefore thought to play a role in radiation enteropathy and can contribute towards radiation induced bowel toxicity. Lactobacilli residing in the gut shares a long conjoined evolutionary history with their hosts and by doing so these organisms have developed an intimate and complex symbiotic relationships. The objective behind this study was to look for the strains with varying resistance to ionizing radiation and to see whether the niche of the bacteria is playing any role in radiation resistance property of bacteria. In this study, we have isolated the Lactobacillus spp. from probiotic preparation and murine gastrointestinal tract, both of which were supposed to be the important source for its isolation. Biochemical characterization did not show a significant difference in the properties, while a significant preference was observed in carbohydrate utilization capacity by the isolates. Effect of ionizing radiations induced by Co60 gamma radiation (10 Gy) on lactobacilli cells was investigated. A cellular survival curve versus absorbed doses was determined. Radiation resistance studies showed that the response of isolates towards cobalt-60 gamma radiation differs from each other and significant decrease in survival was observed in a dose-dependent manner. Thus the present study revealed that the property of radioresistance in Lactobacillus depends upon the source from where they have been isolated.

Keywords: dysbiosis, lactobacillus, mitigation, radiation

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Authors: Ogar Ofut Tumenayu, Olga Shabalina

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This paper describes the application of two types of pedagogical agents’ technology with different functions in an adaptive educational game with the sole aim of improving learning and enhancing interactivities in Digital Educational Games (DEG). This idea could promote the elimination of some problems of DEG, like isolation in game-based learning, by introducing a tutor and tutee pedagogical agents. We present an analysis of a learning companion interacting in a peer tutoring environment as a step toward improving social interactions in the educational game environment. We show that tutor and tutee agents use different interventions and interactive approaches: the tutor agent is engaged in tracking the learner’s activities and inferring the learning state, while the tutee agent initiates interactions with the learner at the appropriate times and in appropriate manners. In order to provide motivation to prevent mistakes and clarity a game task, the tutor agent uses the help dialog tool to provide assistance, while the tutee agent provides collaboration assistance by using the hind tool. We presented our idea on a prototype game called “Pyramid Programming Game,” a 2D game that was developed using Libgdx. The game's Pyramid component symbolizes a programming task that is presented to the player in the form of a puzzle. During gameplay, the Agents can instruct, direct, inspire, and communicate emotions. They can also rapidly alter the instructional pattern in response to the learner's performance and knowledge. The pyramid must be effectively destroyed in order to win the game. The game also teaches and illustrates the advantages of utilizing educational agents such as TrA and TeA to assist and motivate students. Our findings support the idea that the functionality of a pedagogical agent should be dualized into an instructional and learner’s companion agent in order to enhance interactivity in a game-based environment.

Keywords: tutor agent, tutee agent, learner’s companion interaction, agent collaboration

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Authors: Fariha Akhter Chowdhury, Sabrina Mahboob, Anamika Saha, Afrin Jahan, Mohammad Nurul Islam

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Urinary tract infection (UTI) is frequently experienced by the female population where the prevalence increases with aging. Escherichia coli, one of the most common UTI causing organisms, retains glutathione defense mechanism that aids the organism to withstand the harsh physiological environment of urinary tract, host oxidative immune response and even to affect antibiotic-mediated cell death and the emergence of resistance. In this study, we aimed to investigate the glutathione reductase activity of uropathogenic E. coli (UPEC) by observing the reduced glutathione (GSH) level alteration under stressful condition. Urine samples of 58 patients with UTI were collected. Upon isolation and identification, 88% of the samples presented E. coli as UTI causing organism among which randomly selected isolates (n=9), obtained from urine samples of female patients, were considered for this study. E. coli isolates were grown under normal and stressful conditions where H₂O₂ was used as the stress-inducing agent. GSH level estimation of the isolates in both conditions was carried out based on the colorimetric measurement of 5,5'-dithio-bis (2-nitrobenzoic acid) (DTNB) and GSH reaction product using microplate reader assay. The GSH level of isolated E. coli sampled from adult patients decreased under stress compared to normal condition (p = 0.011). On the other hand, GSH production increased markedly in samples that were collected from elderly subjects (p = 0.024). A significant partial correlation between age and change of GSH level was found as well (p = 0.007). This study may help to reveal ways for better understanding of E. coli pathogenesis of UTI prevalence in elderly patients.

Keywords: Escherichia coli, glutathione reductase activity, oxidative stress, reduced glutathione (GSH), urinary tract infection (UTI)

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Authors: Irene Lami, Silvia Micheli, Jan Radimský, Joost van de Weijer

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The present study examines gender agreement in Italian compounds with "capo-". Compounds containing "capo-" as the first element is highly productive in Italian and are attested from the earliest stages of the language, with "capo" indicating a prominent role in a group. This type of compound has become progressively more productive over time, establishing itself in the language to indicate human referents with a leadership role over someone or something belonging to both subordinate and coordinate compound categories. In light of the debates on the use of inclusive language, especially with regard to female professional titles in Italian, the gender agreement of the word "capo" is investigated, which in addition to social resistance, also encounters etymological resistance. Regarding the gender agreement of the word "capo-" as the first element of compounds, in addition to social and etymological resistances, morphological constraints must also be considered. In our experiment, 190 native informants were asked to match the gender of the given the word in a sentence, thinking of female referents. The results confirm a scalar hypothesis of gender agreement (i.e., titles traditionally attributed to women > titles traditionally attributed to men > the word "capo" in isolation > the word "capo-" as an element of subordinate compound > the word “capo-“ as an element of a coordinate compound). A significant interplay with number marking is also shown, as words are inflected in gender when the trait +plural is present. Moreover, the results show that, contrary to what is prescriptively established, speakers do inflect the word "capo" according to gender, in limited instances, even when this is found as a compound element, even though to a lesser extent than words that only have social hinders and not etymological or morphological ones. The results appear to show that, although a morphological obstacle is visible, sociolinguistic claims seem to be able to divert these obstacles. This study appears particularly suitable for replication tests over the next few decades, which, if society opens up further to claims of inclusiveness, could further corroborate this trend.

Keywords: compounds, gender inflection, Italian, morphology

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Authors: Sally Ibrahim, Mohamed Hedia, Mohamed Taqi, Mohamed Derbala, Karima Mahmoud, Youssef Ahmed, Sayed Ismail, Mohamed El-Belely

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The identification and quantification of serum microRNA (miRNA) from mares with endometritis might serve as useful and implementable clinical biomarkers for the early diagnosis of endometiritis. Aims of the current study were (I) to study the expression pattern of eca-miR-155, eca-miR-223, eca-miR-17, eca-miR-200a, and eca-miR-205, and (II) to determine the levels of interleukin 6 (IL-6), prostaglandins (PGF₂α and PGE₂), in the serum of Arabian mares with healthy and abnormal uterine status (endometritis). This study was conducted on 80 Arabian mares (4-14 years old). Mares were divided into 48 sub-fertile mares suspected of endometritis and 32 fertile at stud farms. The criteria for mares to be enrolled in the endometritis group were that they had been bred three or more times unsuccessfully in the breeding season or had a history of more than one year of reproductive failure. In addition, two or more of the following criteria on a checklist were present: abnormal clinical findings, transrectal ultrasonographic uterine examination showed abnormal fluid in the uterus (echogenic or ≥2 cm in diameter), positive endometrial cytology; and bacterial and/or fungal growth. Serum samples were collected for measuring IL-6, PGF₂α, and PGE₂ concentrations, as well as serum miRNA isolation and quantitative real-time PCR. Serum concentrations of IL-6, PGE₂, and PGF₂α were higher (P ≤ 0.001) in mares with endometritis compared to the control healthy ones. The expression profile of eca-miR-155, eca-miR-223, eca-miR-17, eca-miR-200a, and eca-miR-205 increased (P≤0.001) in mares with endometritis compared to the control ones. To the best of our knowledge, this is the first study that revealed that serum miRNA and serum inflammatory mediators (IL-6, PGE₂, and PGF₂α) could be used as non-invasive gold standard biomarkers, and therefore might be served as an important additional diagnostic tool for endometritis in Arabian mares. Moreover, estimation of the serum concentrations of serum miRNA, IL-6, PGE₂, and PGF₂α is a promising recommended tool during the breeding soundness examination in mares.

Keywords: Arabian Mares, endometritis, inflammatory mediators, serum miRNA

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Authors: Hermalina Sinay, Estri L. Arumingtyas

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The research objective was to determine the expression level of dehydration responsive element binding/DREB gene of local corn cultivars from Kisar Island Maluku. The study design was a randomized block design with single factor consist of six local corn cultivars obtained from farmers in Kisar Island and one reference varieties wich has been released by the government as a drought-tolerant varieties and obtained from Cereal Crops Research Institute (ICERI) Maros South Sulawesi. Leaf samples were taken is the second leaf after the flag leaf at the 65 days after planting. Isolation of total RNA from leaf samples was carried out according to the protocols of the R & A-BlueTM Total RNA Extraction Kit and was used as a template for cDNA synthesis. The making of cDNA from total RNA was carried out according to the protocol of One-Step Reverse Transcriptase PCR Premix Kit. Real Time-PCR was performed on cDNA from reverse transcription followed the procedures of Real MODTM Green Real-Time PCR Master Mix Kit. Data obtained from the real time-PCR results were analyzed using relative quantification method based on the critical point / Cycle Threshold (CP / CT). The results of gene expression analysis of DREB gene showed that the expression level of the gene was highest obtained at Deep Yellow local corn cultivar, and the lowest one was obtained at the Rubby Brown Cob cultivar. It can be concluded that the expression level of DREB gene of Deep Yellow local corn cultivar was highest than other local corn cultivars and Srikandi variety as a reference variety.

Keywords: expression, level, DREB gene, local corn cultivars, Kisar Island, Maluku

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Authors: Anastasia Papachristou, Anastasia Ntikoudi, Vasileios Saridakis

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The safety and health care needs of refugees have become an increasingly important issue all over the world especially during last few decades. Wars are the primary reason for refugees to leave their countries. Moreover, refugees are frequently exposed to a variety of stressors such as socioeconomic disadvantages, poverty, changes in family structure and functioning, losing social support, difficulty to access education, living in very crowded places, experiencing racism and isolation. This systematic review included research studies published between 2007-2017 from the search databases Medline, Scopus, Cinahl and PubMed, with keywords 'war survivors', 'war trauma', 'psychiatric disorders', 'refugees'. In order to meet the purpose of the systematic review, further research for complementary studies was conducted into the literature references of the research articles included in this study that would meet the criteria. Overall, 14 studies were reviewed and evaluated. The majority of them demonstrated that the most common psychiatric disorders observed among war refugees are post-traumatic stress disorder (PTSD), depression, anxiety and multiple somatic complaints. Moreover, significant relationship was shown between the number of traumatic events experienced by the refugees and sociodemographic features such as gender, age and previous family history of any psychological disorder. War violence is highly traumatic, causing multiple, long-term negative outcomes such as the aforementioned psychiatric disorders. The number of the studies reviewed in this systematic review is not representative of the problem and its significance. The need for care of the survivors and their families is vital. Further research is necessary in order to clarify the role of predictive factors in the development and maintenance of post-traumatic stress and the rest psychiatric disorders following war trauma. In conclusion, it is necessary to have large multicenter studies in the future in order to be able to draw reliable conclusions about the effects of war.

Keywords: psychiatric disorders, refugees, war survivors, war trauma

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Authors: Indrani Chattopadhyaya

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This paper seeks to examine how identity – a cultural self-image of a group of people develops – how they live, they think, they celebrate and express their world view through language, gesture, symbols, and rituals. 'Culture' is a way of life and 'identity' is assertion of that cultural self-image practiced by the group. The way in which peoples live varies from time to time and from place to place. This variation is important for their identity. Archaeologists have classified these patterns of spacial variations as 'archaeological culture.' These cultures are identified 'self-consciously' with a particular social group indicating ethnicity. The ethnic identity as archaeological cultures also legitimizes the claims of modern groups to territory. In prehistoric research problems of ethnicity and multiculturalism, stylistic attributes significantly reflect both group membership and individuality. In India, anthropologists feel that though tribes have suffered relative isolation through history, they have remained an integral part of Indian civilization. The term 'tribe' calls for substitution with a more meaningful name with an indigenous flavour 'Adivasi' (original inhabitants of the land).While studying prehistoric rock paintings from central India - Sonbhadra (Uttar Pradesh) and Bhimbetka (Madhya Pradesh), one is struck by the similarity between stylistic attributes of painted motifs in the prehistoric rock shelters and the present day indigenous art of Kol and Bhil tribes in the area, who have not seen these prehistoric rock paintings, yet are carrying on with the tradition of painting and decorating their houses in the same way. They worship concretionary sandstone blocks with triangular laminae as Goddess, Devi, Shakti. This practice is going on since Upper Palaeolithic period confirmed by archaeological excavation. The past is legitimizing the role of the present groups by allowing them to trace their roots from earlier times.

Keywords: ethnic identity, hermeneutics, semiotics, Adivasi

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Authors: Sílvia Pereira Mello, Eliane Rodrigues, Maria de Lourdes Andrade, Marcelo Pereira, Giselle Dias, Jose Seixas Filho

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Frog meat is a highly digestible food and its use is recommended in diets aimed at fighting cholesterol, obesity, and arterial hypertension, as well as for treating gastrointestinal disorders. In this study, the soups were developed with frog meat in addition to other ingredients which did not present allergenic potential. The carcasses of the thawed frogs went through bleaching and deboning, and other ingredients (vegetables and condiments) were then added to the separated meat. After the process of cooking, the soups were cooled and later on frozen at -40° C for 3 hours and then taken to the LS 3000 B lyophilizer for 24 hours. The soups were submitted to microbiological analysis: enumeration of total coliforms and Bacillus cereus; identification of coagulase positive Staphylococcus; isolation and identification of Salmonella spp.; and physical-chemical analysis; application of micro-Kjeldahl method for protein, Soxhlet method for lipids, use of a heating chamber at 105ºC for moisture, incineration method (500-550°C) for ash, and Decagon's Pawkit equipment for determining water activity. Acceptance test was performed with 50 elderly people, all between 60 and 85 years of age. The degree of acceptance was demonstrated using a seven points structured hedonic scale in which the taster expressed their impression towards the product. Results of the microbiological analysis showed that all samples met the standards established by the National Health Surveillance Agency of Brazil (ANVISA). Results of the acceptance test indicated that all the soups were accepted considering overall impression and intended consumption. In addition to its excellent nutritional quality, the dehydrated soups made with frog meat are presented as a solution for consumers due to convenience in preparation, consumption and storage.

Keywords: bacteriological quality, lithobates catesbeianus, instant soup, proximate composition, sensory analysis

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Authors: O. I. Adeniran, M. A. Mogale

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Advanced glycation end-products (AGEs) have been implicated in the development and progression of vascular complications of diabetes mellitus and other age-related disease such as Alzheimer’s disease, heart diseases, stroke and limb amputation. The aim of the study was to determine the anti-glycation activity and AGE-cross-linking breaking ability of Sclerocarya birrea stem-bark extracts (SBSBETs). Hexane, ethyl acetate, methanol and water extracts of Sclerocarya birrea stem-bark and standard inhibitor, aminoguanidine (AG) were incubated with bovine serum albumin (BSA)-fructose mixture for 20 and 40 days. The amounts of total immunogenic AGEs (TIAGEs), fluorescent AGEs (FAGEs) and carboxymethyl lysine (CML) formed were determined and the percentage anti-glycation activity of each plant extract calculated. The ability of SBSBETs to break fructose-derived BSA-AGE-collagen cross-links was also investigated. All SBSBETs under investigation demonstrated less anti-glycation activity against TIAGE, FAGEs and CML than AG after 20 days incubation. After 40 days incubation, ethyl acetate, methanol and water SBSBETs demonstrated lower anti-glycation activity against TIAGEs than AG but exerted higher anti-glycation activity than AG against FAGEs. All SBSBETs except water demonstrated lower anti-glycation activity than AG against CML. With regard to the ability of SBSBETs to breakdown fructose-derived AGEs cross-links, the polar SBSBETs demonstrated higher ability to break AGE-cross-links than the non-polar ones. The results of this study may lead to the isolation of bio-active phyto-chemicals from SBSBETs that may be used for the prevention of vascular complication of diabetes.

Keywords: advanced glycation end-products, anti-glycation, cross-link breaking, Sclerocarrya birrea

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Authors: R. S. Komartin, B. Balanuca, R. Stan

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the last decades, studies about the use of polymeric materials of plant origin, considering environmental concerns, have captured the interest of researchers because these represent an alternative to petroleum-derived materials. Vegetable oils are one of the preferred alternatives for petroleum-based raw materials having long aliphatic chains similar to hydrocarbons which means that can be processed using conventional chemistry. Epoxidized vegetable oils (EVO) are among the most interesting products derived from oil both for their high reactivity (epoxy group) and for the potential to react with compounds from various classes. As in the case of epoxy resins starting from petrochemical raw materials, those obtained from EVO can be crosslinked with different agents to build polymeric networks and can also be reinforced with various additives to improve their thermal and mechanical performances. Among the multitude of known EVO, the most common in industrial practice are epoxidized linseed oils (ELO) and epoxidized soybean oils (ESO), the first with an iodine index over 180, the second having a lower iodine index but being cheaper. On the other hand, lignin (Ln) is the second natural organic material as a spread, whose use has long been hampered because of the high costs associated with its isolation and purification. In this context, our goal was to obtain new composite materials with satisfactory intermediate properties in terms of stiffness and elasticity using the characteristics of ELO and Ln and choosing the proper curing procedure. In the present study linseed oil (LO) epoxidation was performed using peracetic acid generated in situ. The obtained bio-based epoxy resin derived from linseed oil was used further to produce the new composites byloading Ln in various mass ratios. The resulted ELO-Ln blends were subjected to a dual-curing protocol, namely photochemical and thermal. The new ELO-Ln composites were investigated by FTIR spectrometry, thermal stability, water affinity, and morphology. The positive effect of lignin regarding the thermal stability of the composites could be proved. The results highlight again the still largely unexplored potential of lignin in industrial applications.

Keywords: composite materials, dual curing, epoxidized linseed oil, lignin

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Authors: Zhanara Suleimenova, Raushan Blieva, Aigerim Zhakipbekova, Inkar Tapenbayeva, Zhanar Narmuratova

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Green processes are based on innovative technologies that do not negatively affect the environment. Industrial enzymes originated from biological systems can effectively contribute to sustainable development through being isolated from microorganisms which are fermented using primarily renewable resources. Many widespread microorganisms secrete a significant amount of biocatalysts into the environment, which greatly facilitates the task of their isolation and purification. The ability to control the enzyme production through the regulation of their biosynthesis and the selection of nutrient media and cultivation conditions allows not only to increase the yield of enzymes but also to obtain enzymes with certain properties. In this regard, large potentialities are embedded in immobilized cells. Enzyme production technology in a secreted active form enabling industrial application on an economically feasible scale has been developed. This method is based on the immobilization of enzyme producers on a solid career. Immobilizing has a range of advantages: decreasing the price of the final product, absence of foreign substances, controlled process of enzyme-genesis, the ability of various enzymes' simultaneous production, etc. Design of proposed equipment gives the opportunity to increase the activity of immobilized cell culture filtrate comparing to free cells, growing in periodic culture conditions. Such technology allows giving a 10-times raise in culture productivity, to prolong the process of fungi cultivation and periods of active culture liquid generation. Also, it gives the way to improve the quality of filtrates (to make them more clear) and exclude time-consuming processes of recharging fermentative vials, that require manual removing of mycelium.

Keywords: industrial enzymes, immobilization, submerged fermentation, microscopic fungi

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Authors: Louise Elizabeth Bolton

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Statement of the problem: There are an estimated 328 million cases of COPD worldwide. It is likely to become the third biggest cause of death by 2030. The impact of living with palliative care needs arising from COPD disrupts an individual’s existential situation. Understandings of individuals' existential situations within COPD are limited within the research literature and are rarely addressed within clinical practice, yet existential suffering has been linked to poor health-related quality of life for those living with other chronic conditions. The purpose of this integrative review is to provide a synthesis of existing evidence on existential suffering for those living with palliative care needs arising from COPD. Methods: This is an integrative review undertaken in accordance with PRISMA guidelines. Nine electronic databases were searched from April 2019 to January 2021. Thirty-five empirical research papers of both qualitative and quantitative methodologies, alongside systematic literature reviews, were included. Data analysis was undertaken using an integrative thematic analysis approach. Findings: Identified themes of existential suffering when living with palliative care needs arising from COPD are as follows: Liminality, Lamented Life, Loss of Personal Liberty, Life Meaning and Existential isolation. The absence of life meaning and purpose was of most importance to patients. Conclusion and Significance: This integrative review provides a synthesis of international evidence upon the presence of existential suffering. It is present and of significant impact within the daily lives of those living with palliative care needs arising from COPD. The absence of life meaning has the most significant impact, requiring further exploration of both its physical and psychological impact. Rediscovery of life meaning diminishes feelings of worthlessness and hopelessness in daily life and facilitates feelings of inner peace. For those with COPD living with such a relentless symptom burden, a positive existential situation is desirable.

Keywords: palliative care, COPD, existential suffering, end of life care

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Authors: Enny Suswati, Supangat Supangat

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Background. Ready-to-eat food products are becoming increasingly popular because consumers are increasingly busy, competitive, and changing lifestyles. Examples of ready-to-eat foods include crispy fried chicken. Escherichia coli is one of the most important causes of food-borne diseases and the most frequent antibiotic-resistant pathogen globally. This study assessed the prevalence and antibiotic resistance profile of E. coli from ready-to-eat crispy fried chicken in Jember city, Indonesia. Methodology. This cross-sectional study was conducted from November 2020 to April 2021 by collecting 81crispy fried chicken samples from 27 food stalls in campus area using a simple random sampling method. Isolation and determination of E. coli use were performed by conventional culture method. An antibiotic susceptibility test was conducted using Kirby Bauer disk diffusion method on the Mueller–Hinton agar. Result. Out of 81crispy fried chicken samples, 77 (95.06%) were positive for E. coli. High E. coli drug resistance was observed on ampicillin, amoxicillin (100%) followed by cefixime (98.72%), erythromycin (97.59%), sulfamethoxazole (93.59%), azithromicin (83.33%), cefotaxime (78.28%), choramphenicol (75.64%), and cefixime (74.36%). On the other hand, there was the highest susceptibility for ciprofloxacin (64.10%). The multiple antibiotic resistance indexes of E. coli isolates varied from 0.4 to 1. The predominant antimicrobial resistance profiles of E. coli were CfmCroAmlAmpAzmCtxSxtCE (n=17), CfmCroAmlCipAmpAzmCtxSxtCE (n=16), and CfmAmlAmpAzmCtxSxtCE (n=5), respectively. Multidrug resistance was also found in the isolates' 76/77 (98.70%). Conclusion. The resistance pattern CfmCroAmlAmpAzmCtxSxtCE was the most common among the E. coli isolates, with 17 showing it. The multiple antibiotic index (MAR index) ranged from 0.4 to 1. Hygienic measures should be rigorously implemented and monitoring resistance of E. coli is required to reduce the risks related to the emergence of multi-resistant bacteria

Keywords: antibacterial drug, ready to eat, crispy fried chicken, escherichia coli

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Authors: Maryam Beiranvand, Sajad Yaghoubi

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Background: Some microbes can colonize plants’ inner tissues without causing obvious damage and can even produce useful bioactive substances. In the present study, the diversity of the endophytic bacteria associated with medicinal plants from Iran was investigated by culturing techniques, molecular gene identification, as well as measuring them for antibacterial activity. Results: In the spring season from 2013 to 2014, 35 herb pharmacology samples were collected, sterilized, meshed, and then cultured on selective media culture. A total of 199 endophytic bacteria were successfully isolated from 35 tissue cultures of medical plants, and sixty-seven out of 199 bacterial isolates were subjected to identification by the 16S rRNA gene sequence analysis method. Based on the sequence similarity gene and phylogenetic analyses, these isolates were grouped into five classes, fourteen orders, seventeen families, twenty-one genera, and forty strains. The most abundant group of endophytic bacteria was actinobacterial, consisting of thirty-two (47%) out of 67 bacterial isolates. Ten (22.3%) out of 67 bacterial isolates remained unidentified and classified at the genus level. The signature of the 16S rRNA gene formed a distinct line in a phylogenetic tree showing that they might be new species of bacteria. One (5.2%) out of 67 bacterial isolates was still not well categorized. Forty-two out of 67 strains were candidates for antimicrobial activity tests. Nineteen (45%) out of 42 strains showed antimicrobial activity multidrug resistance (MDR); thirteen (68%) out of 19 strains were allocated to classes actinobacteria. Four (21%) out of 19 strains belonged to the Bacillaceae family, one (5.2%) out of 19 strains was the Paenibacillaceae family, and one (5.2%) out of 19 strains belonged to the Pseudomonadaceae family. The other twenty-three strains did not show inhibitory activities. Conclusions: Our research showed a high-level phylogenetic diversity and the intoxicating antibiotic activity of endophytic bacteria in the herb pharmacology of Iran.

Keywords: Antibacterial activity, endophytic bacteria, multidrug-resistant bacteria, whole genom sequencing

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