Search results for: bacterial isolation

Authors: Tahmina Huq

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This paper presents the findings of an action research study that aimed to investigate the efficacy of a visual dialogue strategy in assisting autistic students with intellectual disabilities in managing their immediate emotions and improving their academic achievements. The research sought to explore the effectiveness of teaching self-regulation techniques as an alternative to traditional approaches involving segregation. The study identified visual dialogue as a valuable tool for promoting self-regulation in this specific student population. Action research was chosen as the methodology due to its suitability for immediate implementation of the findings in the classroom. Autistic students with intellectual disabilities often face challenges in controlling their emotions, which can disrupt their learning and academic progress. Conventional methods of intervention, such as isolation and psychologist-assisted approaches, may result in missed classes and hindered academic development. This study introduces the utilization of visual dialogue between students and teachers as an effective self-regulation strategy, addressing the limitations of traditional approaches. Action research was employed as the methodology for this study, allowing for the direct application of the findings in the classroom. The study observed two 15-year-old autistic students with intellectual disabilities who exhibited difficulties in emotional regulation and displayed aggressive behaviors. The research question focused on the effectiveness of visual dialogue in managing the emotions of these students and its impact on their learning outcomes. Data collection methods included personal observations, log sheets, personal reflections, and visual documentation. The study revealed that the implementation of visual dialogue as a self-regulation strategy enabled the students to regulate their emotions within a short timeframe (10 to 30 minutes). Through visual dialogue, they were able to express their feelings and needs in socially appropriate ways. This finding underscores the significance of visual dialogue as a tool for promoting emotional regulation and facilitating active participation in classroom activities. As a result, the students' learning outcomes and social interactions were positively impacted. The findings of this study hold significant implications for educators working with autistic students with intellectual disabilities. The use of visual dialogue as a self-regulation strategy can enhance emotional regulation skills and improve overall academic progress. The action research approach outlined in this paper provides practical guidance for educators in effectively implementing self-regulation strategies within classroom settings. In conclusion, the study demonstrates that visual dialogue is an effective strategy for enhancing emotional regulation in autistic students with intellectual disabilities. By employing visual communication, students can successfully regulate their emotions and actively engage in classroom activities, leading to improved learning outcomes and social interactions. This paper underscores the importance of implementing self-regulation strategies in educational settings to cater to the unique needs of autistic students.

Keywords: action research, self-regulation, autism, visual communication

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Authors: Khaled Alabduljabbar

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Background: Benign, isolated, recurrent sixth nerve palsy is very rare in children. Here we report a case of recurrent abducens nerve palsy with no obvious etiology. It is a diagnosis of exclusion. A recurrent benign form of 6th nerve palsy, a rarer still palsy, has been described in the literature, and it is of most likely secondary to inflammatory causes, e.g, following viral and bacterial infections. Purpose: To present a case of 14 months old girl with recurrent attacks of isolated left sixth cranial nerve palsy following upper respiratory tract infection. Observation: The patient presented to opthalmology clinic with sudden onset of inward deviation (esotropia) of the left eye with a compensatory left face turn one week following signs of upper respiratory tract infection. Ophthalmological examination revealed large angle esotropia of the left eye in primary position, with complete limitation of abduction of the left eye, no palpebral fissure changes, and abnormal position of the head (left face turn). Visual acuity was normal, and no significant refractive error on cycloplegic refraction for her age. Fundus examination was normal with no evidence of papilledema. There was no relative afferent pupillary defect (RAPD) and no anisocoria. Past medical history and family history were unremarkable, with no history of convulsion attacks or head trauma. Additional workout include CBC. Erythrocyte sedimentation rate, Urgent magnetic resonance imaging (MRI), and angiography of the brain were performed and demonstrated the absence of intracranial and orbital lesions. Referral to pediatric neurologist was also done and concluded no significant finding. The patient showed improvement of the left sixth cranial nerve palsy and left face turn over a period of two months. Seven months since the first attack, she experienced a recurrent attack of left eye esotropia with left face turn concurrent with URTI. The rest of eye examination was again unremarkable. CT scan and MRI scan of brain and orbit were performed and showed only signs of sinusitis with no intracranial pathology. The palsy resolved spontaneously within two months. A third episode of left 6th nerve palsy occurred 6 months later, whichrecovered over one month. Examination and neuroimagingwere unremarkable. A diagnosis of benign recurrent left 6th cranial nerve palsy was made. Conclusion: Benign sixth cranial nerve palsy is always a diagnosis of exclusion given the more serious and life-threatening alternative causes. It seems to have a good prognosis with only supportive measures. The likelihood of benign 6th cranial nerve palsy to resolve completely and spontaneously is high. Observation for at least 6 months without intervention is advisable.

Keywords: 6th nerve pasy, abducens nerve pasy, recurrent nerve palsy, cranial nerve palsy

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Authors: D. S. N. K. Liyanagamage, V. Karunaratne, A. P. Attanayake, S. Jayasinghe

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Diabetes mellitus is an ever increasing global health problem which causes disability and untimely death. Current treatments using synthetic drugs have caused numerous adverse effects as well as complications, leading research efforts in search of safe and effective alternative treatments for diabetes mellitus. Even though there are traditional Ayurvedic remedies which are effective, due to a lack of scientific exploration, they have not been proven to be beneficial for common use. Hence the aim of this study is to evaluate the traditional remedy made of mixture of plant components, namely leaves of Murraya koenigii L. Spreng (Rutaceae), cloves of Allium sativum L. (Amaryllidaceae), fruits of Garcinia queasita Pierre (Clusiaceae) and seeds of Piper nigrum L. (Piperaceae) used for the treatment of diabetes. We report herein the preliminary results for the in vivo study of the anti-hyperglycaemic activity of the extracts of the above plant mixture in Wistar rats. A mixture made out of equal weights (100 g) of the above mentioned medicinal plant parts were extracted into cold water, hot water (3 h reflux) and water: acetone mixture (1:1) separately. Male wistar rats were divided into six groups that received different treatments. Diabetes mellitus was induced by intraperitoneal administration of streptozotocin at a dose of 70 mg/ kg in male Wistar rats in group two, three, four, five and six. Group one (N=6) served as the healthy untreated and group two (N=6) served as diabetic untreated control and both groups received distilled water. Cold water, hot water, and water: acetone plant extracts were orally administered in diabetic rats in groups three, four and five, respectively at different doses of 0.5 g/kg (n=6), 1.0 g/kg(n=6) and 1.5 g/kg(n=6) for each group. Glibenclamide (0.5 mg/kg) was administered to diabetic rats in group six (N=6) served as the positive control. The acute anti-hyperglycemic effect was evaluated over a four hour period using the total area under the curve (TAUC) method. The results of the test group of rats were compared with the diabetic untreated control. The TAUC of healthy and diabetic rats were 23.16 ±2.5 mmol/L.h and 58.31±3.0 mmol/L.h, respectively. A significant dose dependent improvement in acute anti-hyperglycaemic activity was observed in water: acetone extract (25%), hot water extract ( 20 %), and cold water extract (15 %) compared to the diabetic untreated control rats in terms of glucose tolerance (P < 0.05). Therefore, the results suggest that the plant mixture has a potent antihyperglycemic effect and thus validating their used in Ayurvedic medicine for the management of diabetes mellitus. Future studies will be focused on the determination of the long term in vivo anti-diabetic mechanisms and isolation of bioactive compounds responsible for the anti-diabetic activity.

Keywords: acute antihyperglycemic activity, herbal mixture, oral glucose tolerance test, Sri Lankan medicinal plant extracts

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Authors: Johan Anker

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The purpose of this paper is to discuss the nature of gender construction in modern dystopian fiction, the development of this genre in Young Adult Literature and reasons for the enormous appeal on the adolescent readers. A recent award winning South African text in this genre, The Mark by Edith Bullring (2014), will be used as example while also comparing this text to international bestsellers like Divergent (Roth:2011), The Hunger Games (Collins:2008) and others. Theoretical insights from critics and academics in the field of children’s literature, like Ames, Coats, Bradford, Booker, Basu, Green-Barteet, Hintz, McAlear, McCallum, Moylan, Ostry, Ryan, Stephens and Westerfield will be referred to and their insights used as part of the analysis of The Mark. The role of relevant and recurring themes in this genre, like global concerns, environmental destruction, liberty, self-determination, social and political critique, surveillance and repression by the state or other institutions will also be referred to. The paper will shortly refer to the history and emergence of dystopian literature as genre in adult and young adult literature as part of the long tradition since the publishing of Orwell’s 1984 and Huxley’s Brave New World. Different factors appeal to adolescent readers in the modern versions of this hybrid genre for young adults: teenage protagonists who are questioning the underlying values of a flawed society like an inhuman or tyrannical government, a growing understanding of the society around them, feelings of isolation and the dynamic of relationships. This unease leads to a growing sense of the potential to act against society (rebellion), and of their role as agents in a larger community and independent decision-making abilities. This awareness also leads to a growing sense of self (identity and agency) and the development of romantic relationships. The specific modern tendency of a female protagonist as leader in the rebellion against state and state apparatus, who gains in agency and independence in this rebellion, an important part of the identification with and construction of gender, while being part of the traditional coming-of-age young adult novel will be emphasized. A comparison between the traditional themes, structures and plots of young adult literature (YAL) with adult dystopian literature and those of recent dystopian YAL will be made while the hybrid nature of this genre and the 'sense of unease' but also of hope, as an essential part of youth literature, in the closure to these novels will be discussed. Important questions about the role of the didactic nature of these texts and the political issues and the importance of the formation of agency and identity for the young adult reader, as well as identification with the protagonists in this genre, are also part of this discussion of The Mark and other YAL novels.

Keywords: agency, dystopian literature, gender construction, young adult literature

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Authors: Ruslin, R. E. Kartasasmita, M. S. Wibowo, S. Ibrahim

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An aphrodisiac is a substance contained in food or drug that can arouse sexual instinct and increase pleasure while working, these substances derived from plants, animals, and minerals. When consuming substances that have aphrodisiac activity and duration can improve the sexual instinct. The natural aphrodisiac effect can be obtained through plants, animals, and minerals. Leonurine compound has aphrodisiac activity, these compounds can be isolated from plants of Leonurus Sp, Sundanese people is known as deundereman, this plant is empirical has aphrodisiac activity and based on the isolation of active compounds from plants known to contain compounds leonurine, so that the compound is expected to have activity aphrodisiac. Leonurine compound can be isolated from plants or synthesized chemically with material dasa siringat acid. Leonurine compound can be obtained commercial and derivatives of these compounds can be synthesized in an effort to increase its activity. This study aims to obtain derivatives leonurine better aphrodisiac activity compared with the parent compound, modified the structure of the compounds in the form leonurin guanidino butyl ester group with butyl amin and bromoetanol. ArgusLab program version 4.0.1 is used to determine the binding energy, hydrogen bonds and amino acids involved in the interaction of the compound PDE5 receptor. The in vivo test leonurine compounds and derivatives as an aphrodisiac ingredients and hormone testosterone levels using 27 male rats Wistar strain and 9 female mice of the same species, ages ranged from 12 weeks rats weighing + 200 g / tail. The test animal is divided into 9 groups according to the type of compounds and the dose given. Each treatment group was orally administered 2 ml per day for 5 days. On the sixth day was observed male rat sexual behavior and taking blood from the heart to measure testosterone levels using ELISA technique. Statistical analysis was performed in this study is the ANOVA test Least Square Differences (LSD) using the program Statistical Product and Service Solutions (SPSS). Aphrodisiac efficacy of the leonurine compound and its derivatives have proven in silico and in vivo test, the in silico testing leonurine derivatives have smaller binding energy derivatives leonurine so that activity better than leonurine compounds. Testing in vivo using rats of wistar strain that better leonurine derivative of this compound shows leonurine that in silico studies in parallel with in vivo tests. Modification of the structure in the form of guanidine butyl ester group with butyl amin and bromoethanol increase compared leonurine compound for aphrodisiac activity, testosterone derivatives of compounds leonurine experienced a significant improvement especial is 1RD compounds especially at doses of 100 and 150 mg/bb. The results showed that the compound leonurine and its compounds contain aphrodisiac activity and increase the amount of testosterone in the blood. The compound test used in this study acts as a steroid precursor resulting in increased testosterone.

Keywords: aphrodisiac dysfunction erectile leonurine 1-RD 2-RD, dysfunction, erectile leonurine, 1-RD 2-RD

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Authors: Afsheen Aman, Zainab Bibi, Shah Ali Ul Qader

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Introduction: Xylan is a heteropolysaccharide composed of xylose monomers linked together through 1,4 linkages within a complex xylan network. Owing to wide applications of xylan hydrolytic products (xylose, xylobiose and xylooligosaccharide) the researchers are focusing towards the development of various strategies for efficient xylan degradation. One of the most important strategies focused is the use of heat tolerant biocatalysts which acts as strong and specific cleaving agents. Therefore, the exploration of microbial pool from extremely diversified ecosystem is considerably vital. Microbial populations from extreme habitats are keenly explored for the isolation of thermophilic entities. These thermozymes usually demonstrate fast hydrolytic rate, can produce high yields of product and are less prone to microbial contamination. Another possibility of degrading xylan continuously is the use of immobilization technique. The current work is an effort to merge both the positive aspects of thermozyme and immobilization technique. Methodology: Geobacillus stearothermophilus was isolated from soil sample collected near the blast furnace site. This thermophile is capable of producing thermostable endo-β-1,4-xylanase which cleaves xylan effectively. In the current study, this thermozyme was immobilized within a synthetic and a non-synthetic matrice for continuous production of metabolites using entrapment technique. The kinetic parameters of the free and immobilized enzyme were studied. For this purpose calcium alginate and polyacrylamide beads were prepared. Results: For the synthesis of immobilized beads, sodium alginate (40.0 gL-1) and calcium chloride (0.4 M) was used amalgamated. The temperature (50°C) and pH (7.0) optima of immobilized enzyme remained same for xylan hydrolysis however, the enzyme-substrate catalytic reaction time raised from 5.0 to 30.0 minutes as compared to free counterpart. Diffusion limit of high molecular weight xylan (corncob) caused a decline in Vmax of immobilized enzyme from 4773 to 203.7 U min-1 whereas, Km value increased from 0.5074 to 0.5722 mg ml-1 with reference to free enzyme. Immobilized endo-β-1,4-xylanase showed its stability at high temperatures as compared to free enzyme. It retained 18% and 9% residual activity at 70°C and 80°C, respectively whereas; free enzyme completely lost its activity at both temperatures. The Immobilized thermozyme displayed sufficient recycling efficiency and can be reused up to five reaction cycles, indicating that this enzyme can be a plausible candidate in paper processing industry. Conclusion: This thermozyme showed better immobilization yield and operational stability with the purpose of hydrolyzing the high molecular weight xylan. However, the enzyme immobilization properties can be improved further by immobilizing it on different supports for industrial purpose.

Keywords: immobilization, reusability, thermozymes, xylanase

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Authors: Debjit Ray

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Horizontal gene transfer (HGT) and recombination leads to the emergence of bacterial antibiotic resistance and pathogenic traits. HGT events can be identified by comparing a large number of fully sequenced genomes across a species or genus, define the phylogenetic range of HGT, and find potential sources of new resistance genes. In-depth comparative phylogenomics can also identify subtle genome or plasmid structural changes or mutations associated with phenotypic changes. Comparative phylogenomics requires that accurately sequenced, complete and properly annotated genomes of the organism. Assembling closed genomes requires additional mate-pair reads or “long read” sequencing data to accompany short-read paired-end data. To bring down the cost and time required of producing assembled genomes and annotating genome features that inform drug resistance and pathogenicity, we are analyzing the performance for genome assembly of data from the Illumina NextSeq, which has faster throughput than the Illumina HiSeq (~1-2 days versus ~1 week), and shorter reads (150bp paired-end versus 300bp paired end) but higher capacity (150-400M reads per run versus ~5-15M) compared to the Illumina MiSeq. Bioinformatics improvements are also needed to make rapid, routine production of complete genomes a reality. Modern assemblers such as SPAdes 3.6.0 running on a standard Linux blade are capable in a few hours of converting mixes of reads from different library preps into high-quality assemblies with only a few gaps. Remaining breaks in scaffolds are generally due to repeats (e.g., rRNA genes) are addressed by our software for gap closure techniques, that avoid custom PCR or targeted sequencing. Our goal is to improve the understanding of emergence of pathogenesis using sequencing, comparative genomics, and machine learning analysis of ~1000 pathogen genomes. Machine learning algorithms will be used to digest the diverse features (change in virulence genes, recombination, horizontal gene transfer, patient diagnostics). Temporal data and evolutionary models can thus determine whether the origin of a particular isolate is likely to have been from the environment (could it have evolved from previous isolates). It can be useful for comparing differences in virulence along or across the tree. More intriguing, it can test whether there is a direction to virulence strength. This would open new avenues in the prediction of uncharacterized clinical bugs and multidrug resistance evolution and pathogen emergence.

Keywords: genomics, pathogens, genome assembly, superbugs

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Authors: Benjamin Osei Tutu, Sharon Annison

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On 21^st November, 2016, an outbreak of foodborne illness occurred after a buffet lunch served during a stakeholders’ consultation meeting held in Accra. An investigation was conducted to characterise the affected people, determine the etiologic food, the source of contamination and the etiologic agent and to implement appropriate public health measures to prevent future occurrences. A retrospective cohort study was conducted via telephone interviews, using a structured questionnaire developed from the buffet menu. A case was defined as any person suffering from symptoms of foodborne illness e.g. diarrhoea and/or abdominal cramps after eating food served during the stakeholder consultation meeting in Accra on 21^st November, 2016. The exposure status of all the members of the cohort was assessed by taking the food history of each respondent during the telephone interview. The data obtained was analysed using Epi Info 7. An environmental risk assessment was conducted to ascertain the source of the food contamination. Risks of foodborne infection from the foods eaten were determined using attack rates and odds ratios. Data was obtained from 54 people who consumed food served during the stakeholders’ meeting. Out of this population, 44 people reported with symptoms of food poisoning representing 81.45% (overall attack rate). The peak incubation period was seven hours with a minimum and maximum incubation periods of four and 17 hours, respectively. The commonly reported symptoms were diarrhoea (97.73%, 43/44), vomiting (84.09%, 37/44) and abdominal cramps (75.00%, 33/44). From the incubation period, duration of illness and the symptoms, toxin-mediated food poisoning was suspected. The environmental risk assessment of the implicated catering facility indicated a lack of time/temperature control, inadequate knowledge on food safety among workers and sanitation issues. Limited number of food samples was received for microbiological analysis. Multivariate analysis indicated that illness was significantly associated with the consumption of the snacks served (OR 14.78, P < 0.001). No stool and blood or samples of etiologic food were available for organism isolation; however, the suspected etiologic agent was Staphylococcus aureus or Clostridium perfringens. The outbreak could probably be due to the consumption of unwholesome snack (tuna sandwich or chicken. The contamination and/or growth of the etiologic agent in the snack may be due to the breakdown in cleanliness, time/temperature control and good food handling practices. Training of food handlers in basic food hygiene and safety is recommended.

Keywords: Accra, buffet, conference, C. perfringens, cohort study, food poisoning, gastroenteritis, office workers, Staphylococcus aureus

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Authors: Muna Shalan

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Climate change is a systemic challenge for cities, with its impacts not happening in isolation but rather intertwined, thus increasing hazards and the vulnerability of the exposed population. The increase in the frequency and intensity of heat waves, for example, is associated with multiple repercussions on the quality of life of city inhabitants, including health discomfort, a rise in mortality and morbidity, increasing energy demand for cooling, and shrinking of green areas due to drought. To address the multi-faceted impact of urbanisation-induced warming, municipalities and local governments are challenged with devising strategies and implementing effective response measures. Municipalities are recognising the importance of guiding urban concepts to drive climate action in the urban environment. An example is climate proofing, which refers to a process of mainstreaming climate change into development strategies and programs, i.e., urban planning is viewed through a climate change lens. There is a multitude of interconnected aspects that are critical to paving the path toward climate-proofing of urban areas and avoiding poor planning of layouts and spatial arrangements. Navigating these aspects through an analysis of the overarching practices governing municipal planning processes, which is the focus of this research, will highlight entry points to improve procedures, methods, and data availability for optimising planning processes and municipal actions. By employing a case study approach, the research investigates how municipalities in different contexts, namely in the city of Sahab in Jordan, Chililabombwe in Zambia, and the city of Dortmund in Germany, are integrating guiding urban concepts to shrink the deficit in adaptation and mitigation and achieve climate proofing goals in their respective local contexts. The analysis revealed municipal strategies and measures undertaken to optimize existing building and urban design regulations by introducing key performance indicators and improving in-house capacity. Furthermore, the analysis revealed that establishing or optimising interdepartmental communication frameworks or platforms is key to strengthening the steering structures governing local climate action. The most common challenge faced by municipalities is related to their role as a regulator and implementers, particularly in budget analysis and instruments for cost recovery of climate action measures. By leading organisational changes related to improving procedures and methods, municipalities can mitigate the various challenges that may emanate from uncoordinated planning and thus promote action against urbanisation-induced warming.

Keywords: urbanisation-induced warming, response measures, municipal planning processes, key performance indicators, interdepartmental communication frameworks, cost recovery

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Authors: Marsha D. Loda

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College enrollment is declining and generation Z, today’s college students, are struggling. Before the pandemic, researchers characterized this generational cohort as unique. Gen Z has been called the most achievement-oriented generation, as they enjoy greater economic status, are more racially and ethnically diverse, and better educated than any other generation. However, they are also the most likely generation to suffer from depression and anxiety. Gen Z has grown up largely with usually well-intentioned but overprotective parents who inadvertently kept them from learning life skills, likely impacting their ability to cope with and to effectively manage challenges. The unprecedented challenges resulting from the pandemic up ended their world and left them emotionally reeling. One of the ramifications of this for higher education is how to reengage current Gen Z students in the classroom. This research presents qualitative findings from 24 single-spaced pages of verbatim comments from college students. Research questions concerned what helps them learn and what they abhor, as well as how to engage them with the university outside of the classroom to aid in retention. Students leave little doubt about what they want to experience in the classroom. In order of mention, students want discussion, to engage with questions, to hear how a topic relates to real life and the real world, to feel connections with the professor and fellow students, and to have an opportunity to give their opinions. They prefer a classroom that involves conversation, with interesting topics and active learning. “professor talks instead of lecturing” “professor builds a connection with the classroom” “I am engaged because it feels like a respectful conversation” Similarly, students are direct about what they dislike in a classroom. In order of frequency, students dislike teachers unenthusiastically reading word or word from notes or presentations, repeating the text without adding examples, or addressing how to apply the information. “All lecture. I can read the book myself” “Not taught how to apply the skill or lesson” “Lectures the entire time. Lesson goes in one ear and out the other.” Pertaining to engagement outside the classroom, Gen Z challenges higher education to step outside the box. They don’t want to just hear from professionals in their field, they want to meet and interact with them. Perhaps because of their dependence on technology and pandemic isolation, they seem to reach out for assistance in forming social bonds. “I believe fun and social events are the best way to connect with students and get them involved. Cookouts, raffles, socials, or networking events would all most likely appeal to many students”. “Events… even if they aren’t directly related to learning. Maybe like movie nights… doing meet ups at restaurants”. Qualitative research suggests strategy. This research is rife with strategic implications to improve learning, increase engagement and reduce drop-out rates among Generation Z higher education students. It also compliments existing research on student engagement. With college enrollment declining by some 1.3 million students over the last two years, this research is both timely and important.

Keywords: college enrollment, generation Z, higher education, pandemic, student engagement

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Authors: Salwa Karboune, Amanda Waglay

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Recent studies indicate that health-promoting functional ingredients and nutraceuticals can help support and improve the overall public health, which is timely given the aging of the population and the increasing cost of health care. The development of novel ‘natural’ functional ingredients is increasingly challenging. Biocatalysis offers powerful approaches to achieve this goal. Our recent research has been focusing on the development of innovative biocatalytic approaches towards the isolation of protein isolates from potato by-products and the generation of peptides. Potato is a vegetable whose high-quality proteins are underestimated. In addition to their high proportion in the essential amino acids, potato proteins possess angiotensin-converting enzyme-inhibitory potency, an ability to reduce plasma triglycerides associated with a reduced risk of atherosclerosis, and stimulate the release of the appetite regulating hormone CCK. Potato proteins have long been considered not economically feasible due to the low protein content (27% dry matter) found in tuber (Solanum tuberosum). However, potatoes rank the second largest protein supplying crop grown per hectare following wheat. Potato proteins include patatin (40-45 kDa), protease inhibitors (5-25 kDa), and various high MW proteins. Non-destructive techniques for the extraction of proteins from potato pulp and for the generation of peptides are needed in order to minimize functional losses and enhance quality. A promising approach for isolating the potato proteins was developed, which involves the use of multi-enzymatic systems containing selected glycosyl hydrolase enzymes that synergistically work to open the plant cell wall network. This enzymatic approach is advantageous due to: (1) the use of milder reaction conditions, (2) the high selectivity and specificity of enzymes, (3) the low cost and (4) the ability to market natural ingredients. Another major benefit to this enzymatic approach is the elimination of a costly purification step; indeed, these multi-enzymatic systems have the ability to isolate proteins, while fractionating them due to their specificity and selectivity with minimal proteolytic activities. The isolated proteins were used for the enzymatic generation of active peptides. In addition, they were applied into a reduced gluten cookie formulation as consumers are putting a high demand for easy ready to eat snack foods, with high nutritional quality and limited to no gluten incorporation. The addition of potato protein significantly improved the textural hardness of reduced gluten cookies, more comparable to wheat flour alone. The presentation will focus on our recent ‘proof-of principle’ results illustrating the feasibility and the efficiency of new biocatalytic processes for the production of innovative functional food ingredients, from potato by-products, whose potential health benefits are increasingly being recognized.

Keywords: biocatalytic approaches, functional ingredients, potato proteins, peptides

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Authors: Alan N. A. Heberle, Salatiel W. Da Silva, Valentin Perez-Herranz, Andrea M. Bernardes

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Contaminants of emerging concern are substances widely used, such as pharmaceutical products. These compounds represent risk for both wild and human life since they are not completely removed from wastewater by conventional wastewater treatment plants. In the environment, they can be harm even in low concentration (µ or ng/L), causing bacterial resistance, endocrine disruption, cancer, among other harmful effects. One of the most common taken medicine to treat cardiocirculatory diseases is the Atenolol (ATL), a β-Blocker, which is toxic to aquatic life. In this way, it is necessary to implement a methodology, which is capable to promote the degradation of the ATL, to avoid the environmental detriment. A very promising technology is the advanced electrochemical oxidation (AEO), which mechanisms are based on the electrogeneration of reactive radicals (mediated oxidation) and/or on the direct substance discharge by electron transfer from contaminant to electrode surface (direct oxidation). The hydroxyl (HO•) and sulfate (SO₄•⁻) radicals can be generated, depending on the reactional medium. Besides that, at some condition, the peroxydisulfate (S₂O₈²⁻) ion is also generated from the SO₄• reaction in pairs. Both radicals, ion, and the direct contaminant discharge can break down the molecule, resulting in the degradation and/or mineralization. However, ATL molecule and byproducts can still remain in the treated solution. On this wise, some efforts can be done to implement the AEO process, being one of them the use of a cationic membrane to separate the cathodic (reduction) from the anodic (oxidation) reactor compartment. The aim of this study is investigate the influence of the implementation of a cationic membrane (Nafion®-117) to separate both cathodic and anodic, AEO reactor compartments. The studied reactor was a filter-press, with bath recirculation mode, flow 60 L/h. The anode was an Nb/BDD2500 and the cathode a stainless steel, both bidimensional, geometric surface area 100 cm². The solution feeding the anodic compartment was prepared with ATL 100 mg/L using Na₂SO₄ 4 g/L as support electrolyte. In the cathodic compartment, it was used a solution containing Na₂SO₄ 71 g/L. Between both solutions was placed the membrane. The applied currents densities (iₐₚₚ) of 5, 20 and 40 mA/cm² were studied over 240 minutes treatment time. Besides that, the ATL decay was analyzed by ultraviolet spectroscopy (UV/Vis). The mineralization was determined performing total organic carbon (TOC) in TOC-L CPH Shimadzu. In the cases without membrane, the iₐₚₚ 5, 20 and 40 mA/cm² resulted in 55, 87 and 98 % ATL degradation at the end of treatment time, respectively. However, with membrane, the degradation, for the same iₐₚₚ, was 90, 100 and 100 %, spending 240, 120, 40 min for the maximum degradation, respectively. The mineralization, without membrane, for the same studied iₐₚₚ, was 40, 55 and 72 %, respectively at 240 min, but with membrane, all tested iₐₚₚ reached 80 % of mineralization, differing only in the time spent, 240, 150 and 120 min, for the maximum mineralization, respectively. The membrane increased the ATL oxidation, probably due to avoid oxidant ions (S₂O₈²⁻) reduction on the cathode surface.

Keywords: contaminants of emerging concern, advanced electrochemical oxidation, atenolol, cationic membrane, double compartment reactor

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Authors: Lena Lapidot, Amir Amnon, Rita Nosenko, Veitsman Ella, Cohen-Ezra Oranit, Davidov Yana, Segev Shlomo, Koren Omry, Safran Michal, Ben-Ari Ziv

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Introduction: Hepatocellular carcinoma (HCC) is one of the leading causes of cancer related mortality worldwide. Liver Cirrhosis is the main predisposing risk factor for the development of HCC. The factor(s) influencing disease progression from Cirrhosis to HCC remain unknown. Gut microbiota has recently emerged as a major player in different liver diseases, however its association with HCC is still a mystery. Moreover, there might be an important association between the gut microbiota, nutrition, life style and the progression of Cirrhosis and HCC. The aim of our study was to characterize the gut microbial signature in association with life style and nutrition of patients with Cirrhosis, HCC-Cirrhosis and healthy controls. Design: Stool samples were collected from 95 individuals (30 patients with HCC, 38 patients with Cirrhosis and 27 age, gender and BMI-matched healthy volunteers). All participants answered lifestyle and Food Frequency Questionnaires. 16S rRNA sequencing of fecal DNA was performed (MiSeq Illumina). Results: There was a significant decrease in alpha diversity in patients with Cirrhosis (qvalue=0.033) and in patients with HCC-Cirrhosis (qvalue=0.032) compared to healthy controls. The microbiota of patients with HCC-cirrhosis compared to patients with Cirrhosis, was characterized by a significant overrepresentation of Clostridium (pvalue=0.024) and CF231 (pvalue=0.010) and lower expression of Alphaproteobacteria (pvalue=0.039) and Verrucomicrobia (pvalue=0.036) in several taxonomic levels: Verrucomicrobiae, Verrucomicrobiales, Verrucomicrobiaceae and the genus Akkermansia (pvalue=0.039). Furthermore, we performed an analysis of predicted metabolic pathways (Kegg level 2) that resulted in a significant decrease in the diversity of metabolic pathways in patients with HCC-Cirrhosis (qvalue=0.015) compared to controls, one of which was amino acid metabolism. Furthermore, investigating the life style and nutrition habits of patients with HCC-Cirrhosis, we found significant correlations between intake of artificial sweeteners and Verrucomicrobia (qvalue=0.12), High sugar intake and Synergistetes (qvalue=0.021) and High BMI and the pathogen Campylobacter (qvalue=0.066). Furthermore, overweight in patients with HCC-Cirrhosis modified bacterial diversity (qvalue=0.023) and composition (qvalue=0.033). Conclusions: To the best of the our knowledge, we present the first report of the gut microbial composition in patients with HCC-Cirrhosis, compared with Cirrhotic patients and healthy controls. We have demonstrated in our study that there are significant differences in the gut microbiome of patients with HCC-cirrhosis compared to Cirrhotic patients and healthy controls. Our findings are even more pronounced because the significantly increased bacteria Clostridium and CF231 in HCC-Cirrhosis weren't influenced by diet and lifestyle, implying this change is due to the development of HCC. Further studies are needed to confirm these findings and assess causality.

Keywords: Cirrhosis, Hepatocellular carcinoma, life style, liver disease, microbiome, nutrition

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Authors: L. Abukova, O. Abramova, A. Goreva, Y. Yakovlev

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Important executive decisions on oil and gas production stimulation in Eastern Siberia have been recently taken. There are unique and large fields of oil, gas, and gas-condensate in Eastern Siberia. The Talakan, Koyumbinskoye, Yurubcheno-Tahomskoye, Kovykta, Chayadinskoye fields are supposed to be developed first. It will result in an abrupt increase in environmental load on the nature of Eastern Siberia. In Eastern Siberia, the introduction of ecological imperatives in hydrocarbon production is still realistic. Underground water movement is the one of the most important factors of the ecosystems condition management. Oil and gas production is associated with the forced displacement of huge water masses, mixing waters of different composition, and origin that determines the extent of anthropogenic impact on water drive systems and their protective reaction. An extensive hydrogeological system of the depression type is identified in the pre-salt deposits here. Pressure relieve here is steady up to the basement. The decrease of the hydrodynamic potential towards the basement with such a gradient resulted in reformation of the fields in process of historical (geological) development of the Nepsko-Botuobinskaya anteclise. The depression hydrodynamic systems are characterized by extremely high isolation and can only exist under such closed conditions. A steady nature of water movement due to a strictly negative gradient of reservoir pressure makes it quite possible to use environmentally-harmful liquid substances instead of water. Disposal of the most hazardous wastes is the most expedient in the deposits of the crystalline basement in certain structures distant from oil and gas fields. The time period for storage of environmentally-harmful liquid substances may be calculated by means of the geological time scales ensuring their complete prevention from releasing into environment or air even during strong earthquakes. Disposal of wastes of chemical and nuclear industries is a matter of special consideration. The existing methods of storage and disposal of wastes are very expensive. The methods applied at the moment for storage of nuclear wastes at the depth of several meters, even in the most durable containers, constitute a potential danger. The enormous size of the depression system of the Nepsko-Botuobinskaya anteclise makes it possible to easily identify such objects at the depth below 1500 m where nuclear wastes will be stored indefinitely without any environmental impact. Thus, the water drive system of the Nepsko-Botuobinskaya anteclise is the ideal object for large-volume injection of environmentally harmful liquid substances even if there are large oil and gas accumulations in the subsurface. Specific geological and hydrodynamic conditions of the system allow the production of hydrocarbons from the subsurface simultaneously with the disposal of industrial wastes of oil and gas, mining, chemical, and nuclear industries without any environmental impact.

Keywords: Eastern Siberia, formation pressure, underground water, waste burial

Procedia PDF Downloads 250

Authors: Shahira M. Ezzat, Marwa I. Ezzat, Mona M. Okba, Esther T. Menze, Ashraf B. Abdel-Naim, Shahnas O. Mohamed

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Background: In order to decrease the burden of the high cost of synthetic drugs, it is important to focus on phytopharmaceuticals. The aim of our study was to search for the mechanism of ginger (Zingiber officinale Roscoe) anti-inflammatory potential and to correlate it to its biophytochemicals. Methods: Various extracts viz. water, 50%, 70%, 80%, and 90% ethanol were prepared from ginger rhizomes. Fractionation of the aqueous extract (AE) was accomplished using Diaion HP-20. In vitro anti-inflammatory activity of the different extracts and isolated compounds was evaluated by protein denaturation inhibition, membrane stabilization, protease inhibition, and anti-lipoxygenase assays. In vivo anti-inflammatory activity of AE was estimated by assessment of rat paw oedema after carrageenan injection. Prostaglandin E2 (PGE2), certain inflammation markers (TNF-α, IL-6, IL-1α, IL-1β, INFr, MCP-1MIP, RANTES, and Nox) levels and MPO activity in the paw edema exudates were measured. Total antioxidant capacity (TAC) was also determined. Histopathological alterations of paw tissues were scored. Results: All the tested extracts showed significant (p < 0.1) anti-inflammatory activities. The highest percentage of heat induced albumin denaturation (66%) was exhibited by the 50% ethanol (250 μg/ml). The 70 and 90% ethanol extracts (500 μg/ml) were more potent as membrane stabilizers (34.5 and 37%, respectively) than diclofenac (33%). The 80 and 90% ethanol extracts (500 μg/ml) showed maximum protease inhibition (56%). The strongest anti-lipoxygenase activity was observed for the AE. It showed more significant lipoxygenase inhibition activity than that of diclofenac (58% and 52%, respectively) at the same concentration (125 μg/ml). Fractionation of AE yielded four main fractions (Fr I-IV) which showed significant in vitro anti-inflammatory. Purification of Fr-III and IV led to the isolation of 6-poradol (G1), 6-shogaol (G2); methyl 6- gingerol (G3), 5-gingerol (G4), 6-gingerol (G5), 8-gingerol (G6), 10-gingerol (G7), and 1-dehydro-6-gingerol (G8). G2 (62.5 ug/ml), G1 (250 ug/ml), and G8 (250 ug/ml) exhibited potent anti-inflammatory activity in all studied assays, while G4 and G5 exhibited moderate activity. In vivo administration of AE ameliorated rat paw oedema in a dose-dependent manner. AE (at 200 mg/kg) showed significant reduction (60%) of PGE2 production. The AE at different doses (at 25-200 mg/kg) showed significant reduction in inflammatory markers except for IL-1α. AE (at 25 mg/kg) is superior to indomethacin in reduction of IL-1β. Treatment of animals with the AE (100, 200 mg/kg) or indomethacin (10 mg/kg) showed significant reduction in TNF-α, IL-6, MCP-1, and RANTES levels, and MPO activity by about (31, 57 and 32% ) (65, 60 and 57%) (27, 41 and 28%) (23, 32 and 23%) (66, 67 and 67%) respectively. AE at 100 and 200 mg/kg was equipotent to indomethacin in reduction of NOₓ level and in increasing the TAC. Histopathological examination revealed very few inflammatory cells infiltration and oedema after administration of AE (200 mg/kg) prior to carrageenan. Conclusion: Ginger anti-inflammatory activity is mediated by inhibiting macrophage and neutrophils activation as well as negatively affecting monocyte and leukocyte migration. Moreover, it produced dose-dependent decrease in pro-inflammatory cytokines and chemokines and replenished the total antioxidant capacity. We strongly recommend future investigations of ginger in the potential signal transduction pathways.

Keywords: anti-lipoxygenase activity, inflammatory markers, 1-dehydro-6-gingerol, 6-shogaol

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Authors: Ashir Ahmed

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Peacekeeping is a collective responsibility of governments, law enforcement agencies, communities, families, and individuals. Moreover, the complex nature of peacekeeping activities requires a holistic and collaborative approach where various community sectors work together to form collective strategies that are likely to be more effective than strategies designed and delivered in isolation. Similarly, it is important to learn from past programs to evaluate the initiatives that have worked well and the areas that need further improvement. Review of recent peacekeeping initiatives suggests that there have been tremendous efforts and resources put in place to deal with the emerging threat of terrorism, radicalization and violent extremism through number of de-radicalization programs. Despite various attempts in designing and delivering successful programs for deradicalization, the threat of people being radicalized is growing more than ever before. This research reviews the prominent de-radicalization programs to draw an understanding of their strengths and weaknesses. Some of the weaknesses in the existing programs include. Inaccessibility: Limited resources, geographical location of potential participants (for offline programs), inaccessibility or inability to use various technologies (for online programs) makes it difficult for people to participate in de-radicalization programs. Timeliness: People might need to wait for a program on a set date/time to get the required information and to get their questions answered. This is particularly true for offline programs. Lack of trust: The privacy issues and lack of trust between participants and program organizers are another hurdle in the success of de-radicalization programs. The fear of sharing participants information with organizations (such as law enforcement agencies) without their consent led them not to participate in these programs. Generalizability: Majority of these programs are very generic in nature and do not cater the specific needs of an individual. Participants in these programs may feel that the contents are irrelevant to their individual situations and hence feel disconnected with purpose of the programs. To address the above-mentioned weaknesses, this research developed a framework that recommends some improvements in de-radicalization programs. One of the recommendations is to offer 24/7, secure, private and online hotline (also referred as helpline) for the people who have any question, concern or situation to discuss with someone who is qualified (a counsellor) to deal with people who are vulnerable to be radicalized. To make these hotline services viable and sustainable, the existing organizations offering support for depression, anxiety or suicidal ideation could additionally host these services. These helplines should be available via phone, the internet, social media and in-person. Since these services will be embedded within existing and well-known services, they would likely to get more visibility and promotion. The anonymous and secure conversation between a person and a counsellor would ensure that a person can discuss the issues without being afraid of information sharing with any third party – without his/her consent. The next stage of this project would include the operationalization of the framework by collaborating with other organizations to host de-radicalization hotlines and would assess the effectiveness of such initiatives.

Keywords: de-radicalization, framework, hotlines, peacekeeping

Procedia PDF Downloads 198

Authors: Kirti Hira, A. Sajeli Begum, S. Mahibalan, Poorna Chandra Rao

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Introduction: Cancer is one of the leading causes of death worldwide. Although existing therapy effectively kills cancer cells, they do affect normal growing cells leading to many undesirable side effects. Hence there is need to develop effective as well as safe drug molecules to combat cancer, which is possible through phyto-research. The currently available plant-derived blockbuster drugs are the example for this. In view of this, an investigation was done to identify cytotoxic lead molecules from Hedyotis umbellata (Family Rubiaceae), a widely distributed weed in India. Materials and Methods: The methanolic extract of the whole plant of H. umbellata (MHU), prepared through Soxhlet extraction method was further fractionated with diethyl ether and n-butanol, successively. MHU, ether fraction (EMHU) and butanol fraction (BMHU) were lyophilized and were tested for the cytotoxic effect using 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay against non-small cell lung cancer (NSCLC) A549 cell lines. The potentially active EMHU was subjected to chromatographic purification using normal-phase silica columns, in order to isolate the responsible bioactive compounds. The isolated pure compounds were tested for their cytotoxic effect by MTT assay against A549 cells. Compound-3, which was found to be most active, was characterized using IR, 1H- and 13C-NMR and MS analysis. The study was further extended to decipher the mechanism of action of cytotoxicity of compound-3 against A549 cells through various in vitro cellular models. Cell cycle analysis was done using flow cytometry following PI (Propidium Iodide) staining. Protein analysis was done using Western blot technique. Results: Among MHU, EMHU, and BMHU, the non-polar fraction EMHU demonstrated a significant dose-dependent cytotoxic effect with IC50 of 67.7μg/ml. Chromatography of EMHU yielded seven compounds. MTT assay of isolated compounds explored compound-3 as potentially active one, which inhibited the growth of A549 cells with IC50value of 14.2μM. Further, compound-3 was identified as cedrelopsin, a coumarin derivative having molecular weight of 260. Results of in vitro mechanistic studies explained that cedrelopsin induced cell cycle arrest at G2/M phase and down-regulated the expression of G2/M regulatory proteins such as cyclin B1, cdc2, and cdc25C, dose dependently. This is the first report that explores the cytotoxic mechanism of cedrelopsin. Conclusion: Thus a potential small lead molecule, cedrelopsin isolated from H. umbellata, showing antiproliferative effect mediated by G2/M arrest in A549 cells was discovered. The effect of cedrelopsin against other cancer cell lines followed by in vivo studies can be performed in future to develop a new drug candidate.

Keywords: A549, cedrelopsin, G2/M phase, Hedyotis umbellata

Procedia PDF Downloads 166

Authors: Sima E. Rugarabamu, Mecky I. Matee, Elison N. M. Simon

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Background: In Tanzania bacteriological studies of etiological agents of oro-facial infections are very limited, and very few have investigated anaerobes. The aim of this study was to determine the spectrum of bacterial agents involved in oral and maxillofacial infections in patients attending Muhimbili National Hospital, Dar-es-salaam, Tanzania. Method: This was a hospital based descriptive cross-sectional study that was conducted in the Department of Oral and Maxillofacial Surgery of the Muhimbili National Hospital in Dar es Salaam, Tanzania from 1st January 2014 to 31st August 2014. Seventy (70) patients with various forms of oral and maxillofacial infections who were recruited for the study. The study participants were interviewed using a prepared questionnaire after getting their consent. Pus aspirate was cultured on Blood agar, Chocolate Agar, MacConkey agar and incubated aerobically at 37°C. Imported blood agar was used for anaerobic culture whereby they were incubated at 37°Cin anaerobic jars in an atmosphere of generated using commercial gas-generating kits in accordance with manufacturer’s instructions. Plates were incubated at 37°C for 24 hours (For aerobic culture and 48 hours for anaerobic cultures). Gram negative rods were identified using API 20E while all other isolates were identified by conventional biochemical tests. Antibiotic sensitivity testing for isolated aerobic and anaerobic bacteria was detected by the disk diffusion, agar dilution and E-test using routine and commercially available antibiotics used to treat oral facial infections. Results: This study comprised of 41 (58.5%) males and 29 (41.5%) females with a mean age of 32 years SD +/-15.1 and a range of 19 to 70 years. A total of 161 bacteria strains were isolated from specimens obtained from 70 patients which were an average of 2.3 isolates per patient. Of these 103 were aerobic organism and 58 were strict anaerobes. A complex mix of strict anaerobes and facultative anaerobes accounted for 87% of all infections.The most frequent aerobes isolated was streptococcus spp 70 (70%) followed by Staphylococcus spp 18 (18%). Other organisms such as Klebsiella spp 4 (4%), Proteus spp 5 (5%) and Pseudomonas spp 2 (2%) were also seen. The anaerobic group was dominated by Prevotella spp 25 (43%) followed by Peptostreptococcus spp 18 (31%); other isolates were Pseudomonas spp 2 (1%), black pigmented Pophyromonas spp 4 (5%), Fusobacterium spp 3 (3%) and Bacteroides spp 5 (8%). Majority of these organisms were sensitive to Amoxicillin (98%), Gentamycin (89%), and Ciprofloxacin (100%). A 40% resistance to metronidazole was observed in Bacteroides spp otherwise this drug and others displayed good activity against anaerobes. Conclusions: Oral and maxillofacial facial infections at Muhimbili National Hospital are mostly caused by streptococcus spp and Prevotella spp. Strict anaerobes accounted for 36% of all isolates. The profile of isolates should assist in selecting empiric therapy for infections of the oral and maxillofacial region. Inclusion of antimicrobial agents against anaerobic bacteria is highly recommended.

Keywords: bacteria, oral and maxillofacial infections, antibiotic susceptibility, Tanzania

Procedia PDF Downloads 320

Authors: Victor Pena Ribeiro, Caroline Arruda, Jennyfer Andrea Aldana Mejia, Jairo Kenupp Bastos

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Leishmaniasis is a serious health problem around the world. The treatment of infected individuals with pentavalent antimonial drugs is the main therapeutic strategy. However, they present high toxicity and persistence side effects. Therefore, the discovery of new and safe natural-derived therapeutic agents against leishmaniasis is important. Propolis is a resin of viscous consistency produced by Apis mellifera bees from parts of plants. The main types of Brazilian propolis are green, red, yellow and brown. Thus, the aim of this work was to investigate the chemical composition and leishmanicidal properties of a brown propolis (BP). For this purpose, the hydroalcoholic crude extract of BP was obtained and was fractionated by liquid-liquid chromatography. The chemical profile of the extract and its fractions were obtained by HPLC-UV-DAD. The fractions were submitted to preparative HPLC chromatography for isolation of the major compounds of each fraction. They were analyzed by NMR for structural determination. The volatile compounds were obtained by hydrodistillation and identified by GC/MS. Promastigote forms of Leishmania amazonensis were cultivated in M199 medium and then 2×106 parasites.mL-1 were incubated in 96-well microtiter plates with the samples. The BP was dissolved in dimethyl sulfoxide (DMSO) and diluted into the medium, to give final concentrations of 1.56, 3.12, 6.25, 12.5, 25 and 50 µg.mL⁻¹. The plates were incubated at 25ºC for 24 h, and the lysis percentage was determined by using a Neubauer chamber. The bioassays were performed in triplicate, using a medium with 0.5% DMSO as a negative control and amphotericin B as a positive control. The leishimnicidal effect against promastigote forms was also evaluated at the same concentrations. Cytotoxicity experiments also were performed in 96-well plates against normal (CHO-k1) and tumor cell lines (AGP01 and HeLa) using XTT colorimetric method. Phenolic compounds, flavonoids, and terpenoids were identified in brown propolis. The major compounds were identified as follows: p-coumaric acid (24.6%) for a methanolic fraction, Artepelin-C (29.2%) for ethyl acetate fraction and the compounds of hexane fraction are in the process of structural elucidation. The major volatile compounds identified were β-caryophyllene (10.9%), germacrene D (9.7%), nerolidol (10.8%) and spathulenol (8.5%). The propolis did not show cytotoxicity against normal cell lines (CHO) with IC₅₀ > 100 μg.mL⁻¹, whereas the IC₅₀ < 10 μg.mL⁻¹ showed a potential against the AGP01 cell line, propolis did not demonstrate cytotoxicity against HeLa cell lines IC₅₀ > 100 μg.mL⁻¹. In the determination of the leishmanicidal activity, the highest (50 μg.mL⁻¹) and lowest (1.56 μg.mL⁻¹) concentrations of the crude extract caused the lysis of 76% and 45% of promastigote forms of L. amazonensis, respectively. To the amastigote form, the highest (50 μg.mL⁻¹) and lowest (1.56 μg.mL⁻¹) concentrations caused the mortality of 89% and 75% of L. amazonensis, respectively. The IC₅₀ was 2.8 μg.mL⁻¹ to amastigote form and 3.9 μg.mL⁻¹ to promastigote form, showing a promising activity against Leishmania amazonensis.

Keywords: amastigote, brown propolis, cytotoxicity, promastigote

Procedia PDF Downloads 145

Authors: Shi-Bo Feng, Sheng-Jie Zhang, Jin Zhou

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The relationship between microbial diversity and algal bloom has received considerable attention for decades. Microbes undoubtedly affect annual bloom events and impact the physiology of both partners, as well as shape ecosystem diversity. However, knowledge about interactions and network correlations among broader-spectrum microbes that lead to the dynamics in a complete bloom cycle are limited. In this study, pyrosequencing and network approaches simultaneously assessed the associate patterns among bacteria, archaea, and microeukaryotes in surface water and sediments in response to a natural dinoflagellate (Alexandrium sp.) bloom. In surface water, among the bacterial community, Gamma-Proteobacteria and Bacteroidetes dominated in the initial bloom stage, while Alpha-Proteobacteria, Cyanobacteria, and Actinobacteria become the most abundant taxa during the post-stage. In the archaea biosphere, it clustered predominantly with Methanogenic members in the early pre-bloom period while the majority of species identified in the later-bloom stage were ammonia-oxidizing archaea and Halobacteriales. In eukaryotes, dinoflagellate (Alexandrium sp.) was dominated in the onset stage, whereas multiply species (such as microzooplankton, diatom, green algae, and rotifera) coexistence in bloom collapse stag. In sediments, the microbial species biomass and richness are much higher than the water body. Only Flavobacteriales and Rhodobacterales showed a slight response to bloom stages. Unlike the bacteria, there are small fluctuations of archaeal and eukaryotic structure in the sediment. The network analyses among the inter-specific associations show that bacteria (Alteromonadaceae, Oceanospirillaceae, Cryomorphaceae, and Piscirickettsiaceae) and some zooplankton (Mediophyceae, Mamiellophyceae, Dictyochophyceae and Trebouxiophyceae) have a stronger impact on the structuring of phytoplankton communities than archaeal effects. The changes in population were also significantly shaped by water temperature and substrate availability (N & P resources). The results suggest that clades are specialized at different time-periods and that the pre-bloom succession was mainly a bottom-up controlled, and late-bloom period was controlled by top-down patterns. Additionally, phytoplankton and prokaryotic communities correlated better with each other, which indicate interactions among microorganisms are critical in controlling plankton dynamics and fates. Our results supplied a wider view (temporal and spatial scales) to understand the microbial ecological responses and their network association during algal blooming. It gives us a potential multidisciplinary explanation for algal-microbe interaction and helps us beyond the traditional view linked to patterns of algal bloom initiation, development, decline, and biogeochemistry.

Keywords: microbial community, harmful algal bloom, ecological process, network

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Authors: Mindaugas Liaudanskas, Darius Kviklys, Kristina Zymonė, Raimondas Raudonis, Jonas Viškelis, Norbertas Uselis, Pranas Viškelis, Valdimaras Janulis

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Apples are among the most commonly consumed fruits in the world. Based on data from the year 2014, approximately 84.63 million tons of apples are grown per annum. Apples are widely used in food industry to produce various products and drinks (juice, wine, and cider); they are also used unprocessed. Apples in human diet are an important source of different groups of biological active compounds that can positively contribute to the prevention of various diseases. They are a source of various biologically active substances – especially vitamins, organic acids, micro- and macro-elements, pectins, and phenolic, triterpenic, and other compounds. Triterpenic compounds, which are characterized by versatile biological activity, are the biologically active compounds found in apples that are among the most promising and most significant for human health. A specific analytical procedure including sample preparation and High Performance Liquid Chromatography (HPLC) analysis was developed, optimized, and validated for the detection of triterpenic compounds in the samples of different apples, their peels, and flesh from widespread apple cultivars 'Aldas', 'Auksis', 'Connel Red', 'Ligol', 'Lodel', and 'Rajka' grown in Lithuanian climatic conditions. The conditions for triterpenic compound extraction were optimized: the solvent of the extraction was 100% (v/v) acetone, and the extraction was performed in an ultrasound bath for 10 min. Isocratic elution (the eluents ratio being 88% (solvent A) and 12% (solvent B)) for a rapid separation of triterpenic compounds was performed. The validation of the methodology was performed on the basis of the ICH recommendations. The following characteristics of validation were evaluated: the selectivity of the method (specificity), precision, the detection and quantitation limits of the analytes, and linearity. The obtained parameters values confirm suitability of methodology to perform analysis of triterpenic compounds. Using the optimised and validated HPLC technique, four triterpenic compounds were separated and identified, and their specificity was confirmed. These compounds were corosolic acid, betulinic acid, oleanolic acid, and ursolic acid. Ursolic acid was the dominant compound in all the tested apple samples. The detected amount of betulinic acid was the lowest of all the identified triterpenic compounds. The greatest amounts of triterpenic compounds were detected in whole apple and apple peel samples of the 'Lodel' cultivar, and thus apples and apple extracts of this cultivar are potentially valuable for use in medical practice, for the prevention of various diseases, for adjunct therapy, for the isolation of individual compounds with a specific biological effect, and for the development and production of dietary supplements and functional food enriched in biologically active compounds. Acknowledgements. This work was supported by a grant from the Research Council of Lithuania, project No. MIP-17-8.

Keywords: apples, HPLC, triterpenic compounds, validation

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Authors: Danilo F. Rodrigues, Hérida Regina N. Salgado

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Introduction: The emergence of resistant microorganisms to a large number of clinically approved antimicrobials has been increasing, which restrict the options for the treatment of bacterial infections. As a strategy, drugs with high antimicrobial activities are in evidence. Stands out a class of antimicrobial, the cephalosporins, having as fourth generation cefepime (CEF) a semi-synthetic product which has activity against various Gram-positive bacteria (e.g. oxacillin resistant Staphylococcus aureus) and Gram-negative (e.g. Pseudomonas aeruginosa) aerobic. There are few studies in the literature regarding the development of microbiological methodologies for the analysis of this antimicrobial, so researches in this area are highly relevant to optimize the analysis of this drug in the industry and ensure the quality of the marketed product. The development of microbiological methods for the analysis of antimicrobials has gained strength in recent years and has been highlighted in relation to physicochemical methods, especially because they make possible to determine the bioactivity of the drug against a microorganism. In this context, the aim of this work was the development and validation of a microbiological method for quantitative analysis of CEF in powder lyophilized for injectable solution by turbidimetric assay. Method: For performing the method, Staphylococcus aureus ATCC 6538 IAL 2082 was used as the test microorganism and the culture medium chosen was the Casoy broth. The test was performed using temperature control (35.0 °C ± 2.0 °C) and incubated for 4 hours in shaker. The readings of the results were made at a wavelength of 530 nm through a spectrophotometer. The turbidimetric microbiological method was validated by determining the following parameters: linearity, precision (repeatability and intermediate precision), accuracy and robustness, according to ICH guidelines. Results and discussion: Among the parameters evaluated for method validation, the linearity showed results suitable for both statistical analyses as the correlation coefficients (r) that went 0.9990 for CEF reference standard and 0.9997 for CEF sample. The precision presented the following values 1.86% (intraday), 0.84% (interday) and 0.71% (between analyst). The accuracy of the method has been proven through the recovery test where the mean value obtained was 99.92%. The robustness was verified by the parameters changing volume of culture medium, brand of culture medium, incubation time in shaker and wavelength. The potency of CEF present in the samples of lyophilized powder for injectable solution was 102.46%. Conclusion: The turbidimetric microbiological method proposed for quantification of CEF in lyophilized powder for solution for injectable showed being fast, linear, precise, accurate and robust, being in accordance with all the requirements, which can be used in routine analysis of quality control in the pharmaceutical industry as an option for microbiological analysis.

Keywords: cefepime hydrochloride, quality control, turbidimetric assay, validation

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Authors: Noel Smith

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To achieve professional registration, UK social workers need to complete a programme of education and training which meets standards set down by central government. When it comes to practice, social work in local authorities must fulfil requirements of national legislation but there is considerable local variation in the organisation and delivery of services. This presentation discusses the on-going reform of social work education by central government in the context of research of social work services in a local authority. In doing so it highlights that the ‘direction of travel’ of the national reform of social work education seems at odds with the trajectory of development of local social work services. In terms of education reform, the presentation cites key government initiatives including the knowledge and skills requirements which have been published separately for, respectively, child and family social work and adult social work. Also relevant is the Government’s new ‘teaching partnership’ pilot which focuses exclusively on social work in local government, in isolation from social work in NGOs. In terms of research, the presentation discusses two studies undertaken by Professor Smith in Suffolk County Council, a local authority in the east of England. The first is an equality impact analysis of the introduction of a new model for the delivery of adult and community services in Suffolk. This is based on qualitative research with local government representatives and NGOs involved in social work with older people and people with disabilities. The second study is an on-going, mixed method evaluation of the introduction of a new model of social care for children and young people in Suffolk. This new model is based on the international ‘Signs of Safety’ approach, which is applied in this model to a wide range of services from early intervention to child protection. While both studies are localised, the service models they examine are good illustrations of the way services are developing nationally. Analysis of these studies suggest that, if services continue to develop as they currently are, then social workers will require particular skills which are not be adequately addressed in the Government’s plans for social work education. Two issues arise. First, education reform concentrates on social work within local government while increasingly local authorities are outsourcing service provision to NGOs, expecting greater community involvement in providing care, and integrating social care with health care services. Second, education reform focuses on the different skills required for working with older and disabled adults and working with children and families, to the point where potentially the profession would be fragmented into two different classes of social worker. In contrast, the development of adult and children’s services in local authorities re-asserts the importance of common social work skills relating to personalisation, prevention and community development. The presentation highlights the importance for social work education in the UK to be forward looking, in terms of the changing design of service delivery, and outward looking, in terms of lessons to be drawn from international social work.

Keywords: adult social work, children and families social work, European social work, social work education

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Authors: Pule P. Mpopetsi, Warren M. Potts, Nicola James, Amber Childs

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Increased CO₂ production and the consequent ocean acidification (OA) have been identified as one of the greatest threats to both calcifying and non-calcifying marine organisms. Traditionally, marine fishes, as non-calcifying organisms, were considered to have a higher tolerance to near-future OA conditions owing to their well-developed ion regulatory mechanisms. However, recent studies provide evidence to suggest that they may not be as resilient to near-future OA conditions as previously thought. In addition, earlier life stages of marine fishes are thought to be less tolerant than juveniles and adults of the same species as they lack well-developed ion regulatory mechanisms for maintaining homeostasis. This study focused on the effects of near-future OA on larval Argyrosomus japonicus, an estuarine-dependent marine fish species, in order to identify the tipping point of tolerance for the larvae of this species. Larval A. japonicus in the present study were reared from the egg up to 22 days after hatching (DAH) under three treatments. The three treatments, (pCO₂ 353 µatm; pH 8.03), (pCO₂ 451 µatm; pH 7.93) and (pCO₂ 602 µatm; pH 7.83) corresponded to levels predicted to occur in year 2050, 2068 and 2090 respectively under the Intergovernmental Panel on Climate Change (IPCC) Representative Concentration Pathways (IPCC RCP) 8.5 model. Size-at-hatch, growth, development, and metabolic responses (standard and active metabolic rates and metabolic scope) were assessed and compared between the three treatments throughout the rearing period. Five earlier larval life stages (hatchling – flexion/post-flexion) were identified by the end of the experiment. There were no significant differences in size-at-hatch (p > 0.05), development or the active metabolic (p > 0.05) or metabolic scope (p > 0.05) of fish in the three treatments throughout the study. However, the standard metabolic rate was significantly higher in the year 2068 treatment but only at the flexion/post-flexion stage which could be attributed to differences in developmental rates (including the development of the gills) between the 2068 and the other two treatments. Overall, the metabolic scope was narrowest in the 2090 treatment but varied according to life stage. Although not significantly different, metabolic scope in the 2090 treatment was noticeably lower at the flexion stage compared to the other two treatments, and the development appeared slower, suggesting that this could be the stage most prone to OA. The study concluded that, in isolation, OA levels predicted to occur between 2050 and 2090 will not negatively affect size-at-hatch, growth, development, and metabolic responses of larval A. japonicus up to 22 DAH (flexion/post-flexion stage). The present study also identified the tipping point of tolerance (where negative impacts will begin) in larvae of the species to be between the years 2090 and 2100.

Keywords: climate change, ecology, marine, ocean acidification

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Authors: Nurten Çelik

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Under the new developments in science, medicine, sociology, psychology and literary theories, body studies has gained huge importance and the body has become a debatable issue. There has emerged, among sociologists and literary theorists, an overwhelming consensus that body is socially, politically and culturally perceived and constructed and thus, the position of an individual in the society is determined in accordance with his/her body image. In this regard, the most complicated point is the theoretical views propounded upon disability studies, where the disabled body is considered to be a site upon which social and political restrictions as well as repressions are inscribed. There has been the widely-accepted view that no matter what kind of disability it is, those with physical, mental or learning impairments face varied social, political and environmental obstacles that prevent them from being an active citizen, worker, lover and even a family member. In parallel with these approaches, the matter of the sufferings of disabled individuals attains its place in cinema and literature as well as in theatre studies under the category of disability theatre. One of the prominent plays that deal with physical disability came from the contemporary British playwright Nina Raine. In her awarded play Tribes, which premiered at the Royal Court Theatre in 2010, Raine develops the social strata where her deaf protagonist, Billy, caught up between two tribes – namely his family and his lover Slyvia, a member of the deaf community– experiences personal and social hardships due to his hearing impairment. In the play, intransigent and self-opinionated family members foster no sense of empathy towards Billy, there are noisy talking and shouting, but no communication, love, compassion or mutual understanding, and language becomes just a tool for the expression of rage and oppression. In the disordered atmosphere of the family life, Billy experiences isolation and loneliness. Billy’s hopes for success and love are destroyed when Slyvia, troubled between hearing and deafness, rejects him because she does not utterly grasp what Billy is experiencing. Drawing upon the hardships, Billy undergoes in his relationships with his family and his girlfriend, Tribes problematizes the concept of deafness and explores to what extent a deaf person can find a place in the hearing world. Setting ‘the disabled’ bodies against ‘the abled’ bodies in a family, a microcosm of the society where bodies are socially shaped and constructed, Tribes dramatizes how the disabled bodies are disenfranchised, stigmatised, marginalized and othered on the grounds that they are socially misfit. Tribes, with a specific focus on the dysfunctional family, shows that the lack of communication and empathy numbs the characters to the feelings of each other and thereby, they become more disabled than Billy. In conclusion, this paper, with the reference to the embodiment of disability and social theories, aims to explore how disabled bodies are socially marked and segregated from family and society.

Keywords: body, deafness, disability, disability theatre, Nina Raine, tribes

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Authors: Marina Sofia, Alexios Giannakopoulos, Antonia Touloudi, Dimitris C Chatzopoulos, Zoi Athanasakopoulou, Vassiliki Spyrou, Charalambos Billinis

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Fertility of dairy cattle can be affected by heat stress when the ambient temperature increases above 30°C and the relative humidity ranges from 35% to 50%. The present study was conducted on dairy cattle farms during summer months in Greece and aimed to identify the serological profile against pathogens that could affect fertility and to associate the positive serological results at herd level with temperature factors. A total of 323 serum samples were collected from clinically healthy dairy cows of 8 herds, located in Central and Northern Greece. ELISA tests were performed to detect antibodies against selected pathogens that affect fertility, namely Chlamydophila abortus, Coxiella burnetii, Neospora caninum, Toxoplasma gondii and Infectious Bovine Rhinotracheitis Virus (IBRV). Eleven climatic variables were derived from the WorldClim version 1.4. and ArcGIS V.10.1 software was used for analysis of the spatial information. Five different MaxEnt models were applied to associate the temperature variables with the locations of seropositive Chl. abortus, C. burnetii, N. caninum, T. gondii and IBRV herds (one for each pathogen). The logistic outputs were used for the interpretation of the results. ROC analyses were performed to evaluate the goodness of fit of the models’ predictions. Jackknife tests were used to identify the variables with a substantial contribution to each model. The seropositivity rates of pathogens varied among the 8 herds (0.85-4.76% for Chl. abortus, 4.76-62.71% for N. caninum, 3.8-43.47% for C. burnetii, 4.76-39.28% for T. gondii and 47.83-78.57% for IBRV). The variables of annual temperature range, mean diurnal range and maximum temperature of the warmest month gave a contribution to all five models. The regularized training gains, the training AUCs and the unregularized training gains were estimated. The mean diurnal range gave the highest gain when used in isolation and decreased the gain the most when it was omitted in the two models for seropositive Chl.abortus and IBRV herds. The annual temperature range increased the gain when used alone and decreased the gain the most when it was omitted in the models for seropositive C. burnetii, N. caninum and T. gondii herds. In conclusion, antibodies against Chl. abortus, C. burnetii, N. caninum, T. gondii and IBRV were detected in most herds suggesting circulation of pathogens that could cause infertility. The results of the spatial analyses demonstrated that the annual temperature range, mean diurnal range and maximum temperature of the warmest month could affect positively the possible pathogens’ presence. Acknowledgment: This research has been co‐financed by the European Regional Development Fund of the European Union and Greek national funds through the Operational Program Competitiveness, Entrepreneurship and Innovation, under the call RESEARCH–CREATE–INNOVATE (project code: T1EDK-01078).

Keywords: dairy cows, seropositivity, spatial analysis, temperature factors

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Authors: Sarah Faulkner, Patrick Faulkner, Caroline Ellison

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Improved mental and physical health, community connection, and increased life satisfaction has been strongly associated with bike riding for those with and without a disability. However, much evidence suggests that people living with a disability face increased barriers to engaging in cycling compared to members of the general population. People with an intellectual disability often live more sedentary and socially isolated lives that negatively impact their mental and physical health, as well as life satisfaction. This paper is based on preliminary findings from a three-year intervention cycling project funded by the South Australian Government. The cycling project was developed in partnership with community stakeholders that provided weekly instruction, training, and support to individuals living with intellectual disabilities to increase their capacity in cycling. This project aimed to support people living with intellectual disabilities to foster and facilitate improved physical and mental health, confidence, and independence and enhance social networking through their engagement in community cycling. The program applied principles of social role valorisation (SRV) theory as its guiding framework. Preliminary data collected is based on qualitative interviews with over 50 program participants, results from two participant wellness questionnaires, as well as a perceptually regulated exercise test administered throughout the project implementation. Preliminary findings are further supplemented with ethnographic analyses by the researchers who took a phenology of life experience approach. Preliminary findings of the program suggest a variety of social motivations behind participants' desire to learn cycling that acknowledges previous barriers to engagement and cycling’s role to address feelings of loneliness and social isolation. Meaningful health benefits can be achieved as demonstrated by increases in predicted V02 max measures, suggesting that physical intervention can not only improve physical health outcomes but also provide a variety of other social benefits. Initial engagement in the project has demonstrated an increase in participants' sense of confidence, well-being, and physical fitness. Implementation of the project in partnership with a variety of community stakeholders has identified a number of critical factors and processes necessary for future service replication, sustainability, and success. Findings from this intervention study contribute to the development of a knowledge base on how best to support individuals living with an intellectual disability to partake in bike riding and increase positive outcomes associated with their capacity building, social interaction, increased physical activity, physical health, and mental well-being. The initial findings of this study provide critical academic insights into the social and physical benefits of cycling for people living with a disability, as well as practical advice for future human service applications.

Keywords: cycling, disability, social inclusion, capacity building

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Authors: Leo Nnamdi Ozurumba-Dwight

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Single-cell genomic analytical systems have proved to be a platform to isolate bulk cells into selected single cells for genomic, proteomic, and related metabolomic studies. This is enabling systematic investigations of the level of heterogeneity in a diverse and wide pool of cell populations. Single cell technologies, embracing techniques such as high parameter flow cytometry, single-cell sequencing, and high-resolution images are playing vital roles in these investigations on messenger ribonucleic acid (mRNA) molecules and related gene expressions in tracking the nature and course of disease conditions. This entails targeted molecular investigations on unit cells that help us understand cell behavoiur and expressions, which can be examined for their health implications on the health state of patients. One of the vital good sides of single-cell RNA sequencing (scRNA seq) is its probing capacity to detect deranged or abnormal cell populations present within homogenously perceived pooled cells, which would have evaded cursory screening on the pooled cell populations of biological samples obtained as part of diagnostic procedures. Despite conduction of just single-cell transcriptome analysis, scRNAseq now permits comparison of the transcriptome of the individual cells, which can be evaluated for gene expressional patterns that depict areas of heterogeneity with pharmaceutical drug discovery and clinical treatment applications. It is vital to strictly work through the tools of investigations from wet lab to bioinformatics and computational tooled analyses. In the precise steps for scRNAseq, it is critical to do thorough and effective isolation of viable single cells from the tissues of interest using dependable techniques (such as FACS) before proceeding to lysis, as this enhances the appropriate picking of quality mRNA molecules for subsequent sequencing (such as by the use of Polymerase Chain Reaction machine). Interestingly, scRNAseq can be deployed to analyze various types of biological samples such as embryos, nervous systems, tumour cells, stem cells, lymphocytes, and haematopoietic cells. In haematopoietic cells, it can be used to stratify acute myeloid leukemia patterns in patients, sorting them out into cohorts that enable re-modeling of treatment regimens based on stratified presentations. In immunotherapy, it can furnish specialist clinician-immunologist with tools to re-model treatment for each patient, an attribute of precision medicine. Finally, the good predictive attribute of scRNAseq can help reduce the cost of treatment for patients, thus attracting more patients who would have otherwise been discouraged from seeking quality clinical consultation help due to perceived high cost. This is a positive paradigm shift for patients’ attitudes primed towards seeking treatment.

Keywords: immunotherapy, transcriptome, re-modeling, mRNA, scRNA-seq

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Authors: Safak Akdemir

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The main purpose of this research is to evaluate the relationships between childhood traumas and dissociative experiences, self-perceptions and early maladaptive schemas in male convicts convicted of sexual assault crimes in prison. In our study, male convicts in prison for the crime of sexual assault constitute the experimental group, and the participants matched with this experimental group in terms of education, age and gender constitute the control group. The experimental group of the research consists of 189 male individuals who are convicted in the Ministry of Justice, General Directorate of Prisons, Istanbul/Maltepe L Type Closed Prison. The control group of this study consists of 147 adult males matched with the experimental group in terms of age, gender and education parameters. A total of 336 adult male individuals are included in the sample of this study. 46% of the experimental group were convicted of only sexual assault, 54% of them were convicted of both sexual assault and murder, injury and drug crimes. Total of five data collection tools, namely the Personal Information Form created by S. A. & E. O., Childhood Trauma Questionnaire (CTQ), the Dissociative Experiences Scale (DES), the Rosenberg Self-Esteem Scale (RSES), and the Young Schema Questionnaire-Short Form (YSQ-SF3), were completed. DES cut-off score of 99 (52.39%) of 189 convicts in the experimental group and 12 (8.17%) of 147 people in the control group was found to be 30 and above, and this result indicates the presence of pathological dissociative experiences. 180 (95.23%) of the sexual assault convicts in the experimental group had at least one childhood trauma, 154 (81.48%) were emotional neglect, 140 (74.07%) were emotional abuse, 121 (64.02%) were physical neglect, 91 (4814%) physical abuse and 70 (37.03%) sexual abuse. 168 (88.88%) of the experimental group reported multiple type of trauma and 12 (6.34%) reported single type of trauma. While the childhood traumas, isolation, abandonment and emotional deprivation schema levels of the convicts with a DES cut-off score of 30 and above are higher than the convicts with a DES cut-off score of 30 and above, their self-esteem is lower than this group. Experimental group while childhood traumas, dissociative experiences and early maladaptive schemas are higher than the control group, their self-esteem levels are lower. Dissociative experiences, abandonment and emotional deprivation early maladaptive schemas are more common in convicts aged between 18-30 years compared to convicts aged 31 and over. In addition, dissociative experiences and early maladaptive schemas of male convicts who reported physical and sexual abuse were higher than those who did not report physical and sexual abuse, while their self-esteem was at a lower level. As a result, in terms of psychotraumatology and clinical forensic psychology, dissociative disorders developed under the influence of chronic childhood traumas, with clinical interviews and psychometric measurements to be made in terms of forensic psychiatry; it is of fundamental importance to evaluate it in terms of neurosis-psychosis distinction, disability retirement, custody, malpractice, criminal and legal capacity criteria.

Keywords: crime, sexual assault, criminology, rape crimes, dissocitative disorders, maladative schemas

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Authors: N. G. Klivleyeva, T. I. Glebova, G. V. Lukmanova, S. B. Bayseit, S. Z. Taubaeva, M. K. Kalkozhaeva

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The role of viral diseases in the general infectious disease incidence increases every year and requires special attention to the problem of interpreting the etiology of infectious agents. Influenza and acute respiratory viral infections are one of the most pressing public health issues. In the period 2012-2014, collection of 419 nasal swabs and 150 blood sera has been carried out in the patient care institutions of the various Kazakhstan regions from patients with symptoms of ARVI and pneumonia. Primary identification of biosamples for the presence of influenza viral antigens in enzyme immunoassay on nitrocellulose membrane gave positive results in 125 swabs (29.8%). Biosample screening in immunofluorescence test revealed the presence of influenza viral antigens against A/H1 in 63 samples (15.0%), A/H3 – in 70 samples (16.7%) and type B – in 9 samples (2.1%). As a result of primary infection, and successive passages in chick embryos and MDCK cell cultures, 38 HAAg were isolated from 419 samples with a clear cytopathic effect and hemagglutination titre in MDCK cell culture within 1:2-1:4, in CE - 1:8-1:256. The infectivity of isolates in chicken embryos were 3.5-6.5 lg EID50/0.2, in MDCK cell culture – 2.5-6.5 lg PFU/ml. Identification of 28 isolates was carried out in inhibition reactions of hemagglutinating activity and neuraminidase activity, showed their belonging to the influenza virus: 26 strains to A/H1N1, one - to A/H3N2, and one - to type B. Serological examination of blood sera for the presence of specific antibodies being an indirect evidence of the performed isolation and contributing to the timely interpretation of the disease etiology in the epidemics takes an important place in the comprehensive study of influenza viruses circulating among people. Serological analyzes were carried out in HAI assay using a kit consisting of 12 reference strains obtained from the WHO centre for reference and research on Influenza (CDC, Atlanta, USA) and three Kazakhstan (A/Almaty/347/09 (H1N1v), A/Almaty/462/11 (H3N2) and B/Almaty/414/10) human influenza viruses that are stored in the laboratory collection. The results of serological analysis of 150 blood sera showed that antihaemagglutinins against the A/H3N2 virus serosubtype were found in 46 samples (49.4%) out of 93 sera collected in 2012-2013. The antibody titres were within 1:160-1:320. 19 sera (20.4%) were seropositive against influenza A/H1N1 virus, the antibodies were observed in titres of 1:20-1:40. Six sera (6.4%) were positive against the influenza A/H1N1+A/H3N2 virus (mixed infection); the antibodies were recorded in titres of 1:20-1:40. Antihaemagglutinins against influenza type B virus were detected only in five sera (5.4%). The results of analysis of 57 sera collected in 2014 showed that antihaemagglutinins against A/H3N2 virus subtype were detected in 32 blood sera (56.1%) in titres of 1:160-1:640. Ten sera (17.5%) were seropositive against A/H1N1 virus; antihaemagglutinins against influenza type B virus were not detected. Therefore, virological and serological studies have shown that in Kazakhstan, as well as in the world, the influenza viruses A/H1N1, A/H3N2 and influenza B viruses were actively circulating during the epidemic seasons in 2012-2014.

Keywords: influenza, MDCK cell, serological analysis, virus

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