Search results for: gene family
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 4417

Search results for: gene family

1627 Arbuscular Mycorrhizal Symbiosis in Trema orientalis: Effect of a Naturally-Occurring Symbiosis Receptor Kinase Mutant Allele

Authors: Yuda Purwana Roswanjaya, Wouter Kohlen, Rene Geurts

Abstract:

The Trema genus represents a group of fast-growing tropical tree species within the Cannabaceae. Interestingly, five species nested in this lineage -known as Parasponia- can establish rhizobium nitrogen-fixing root nodules, similar to those found in legumes. Parasponia and legumes use a conserved genetic network to control root nodule formation, among which are genes also essential for mycorrhizal symbiosis (the so-called common symbiotic pathway). However, Trema species lost several genes that function exclusively in nodulation, suggesting a loss-of the nodulation trait in Trema. Strikingly, in a Trema orientalis population found in Malaysian Borneo we identified a truncated SYMBIOSIS RECEPTOR KINASE (SYMRK) mutant allele lacking a large portion of the c-terminal kinase domain. In legumes this gene is essential for nodulation and mycorrhization. This raises the question whether Trema orientalis can still be mycorrhized. To answer this question, we established quantitative mycorrhization assay for Parasponia andersonii and Trema orientalis. Plants were grown in closed pots on half strength Hoagland medium containing 20 µM potassium phosphate in sterilized sand and inoculated with 125 spores of Rhizopagus irregularis (Agronutrion-DAOM197198). Mycorrhization efficiency was determined by analyzing the frequency of mycorrhiza (%F), the intensity of the mycorrhizal colonization (%M) and the arbuscule abundance (%A) in the root system. Trema orientalis RG33 can be mycorrhized, though with lower efficiency compared to Parasponia andersonii. From this we conclude that a functional SYMRK kinase domain is not essential for Trema orientalis mycorrhization. In ongoing experiments, we aim to investigate the role of SYMRK in Parasponia andersonii mycorrhization and nodulation. For this two Parasponia andersonii symrk CRISPR-Cas9 mutant alleles were created. One mimicking the TorSYMRKRG33 allele by deletion of exon 13-15, and a full Parasponia andersonii SYMRK knockout.

Keywords: endomycorrhization, Parasponia andersonii, symbiosis receptor kinase (SYMRK), Trema orientalis

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1626 In vitro and in vivo Potential Effect of the N-Acylsulfonamide Bis-oxazolidin-2-ones on Toxoplasma gondii

Authors: Benlaifa Meriem, Berredjem Hajira, Bouasla Radia, Berredjem Malika, Djebar Med Reda

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Toxoplasmosis is a cosmopolitan infection due to Toxoplasma gondii (T.gondii). It is a significant cause of congenital disease and an important opportunistic pathogen which has become a worldwide increasing problem due to the AIDS epidemic. Current available drugs do not give satisfactory results and often have only a static and several adverse side effects as it is the case of pyrimethamine. So, the need to develop and evaluate new drugs is critical. The purpose of this study is to investigate the in vitro and in vivo effects of the new chiral N-acylsulfonamide bis-oxazolidin-2-ones on T.gondii. In this study, anti-T.gondii RH strain activities, of two new chiral N-acylsulfonamide bis-oxazolidin-2-ones were evaluated in vitro, using a MRC-5 fibroblast tissue cultures to determine the concentration that inhibit parasite multiplication by 50% (IC50) of each drug and in vivo, by PCR detection of the tachyzoites in mice ascites after new molecules treatment, using the 35-fold repetitive B1 gene of T.gondii. The in vitro results demonstrated that the treatment with the tested molecules decreased the amount of tachyzoites in cell culture in a dose-dependent manner. The inhibition was complete for concentrations over 4 mg/ml. The IC50 of Mol 1 and Mol 2 were 1.5 and 3 mg/ml, respectively, and were quite similar to the control one (2 mg/ml). The Mol 1 was highly active against T.gondii in cell cultures than Mol 2; these results were similar to those of sulfadiazine-treated group (p < 0.05). Toxoplasma-specific DNA was demonstrated in all ascites samples from infected mice of the different tested groups. Mol 1 showed better effect than Mol 2, but it did not completely inhibit the parasite proliferation. The intensity of amplification products increased when the treatment started late after infection. These findings suggest continuous parasite replication despite the treatment. In conclusion, our results showed a promising treatment effect of the tested molecules and suggest that in vitro, the Mol 1, and Mol 2 have a dose-dependent effect and a high cytotoxicity on the studied cells. The present study revealed that concentration and duration of tested molecules treatment are major factors that influence the course of Toxoplasma infection in infected mice.

Keywords: cytotoxicity, PCR, sulfonamide, Toxoplasma gondii

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1625 Investigation p53 Codon 72 Polymorphism and miR-146a rs2910164 Polymorphism in Breast Cancer

Authors: Marjan Moradi Fard, Hossein Rassi, Masoud Houshmand

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Aim: Breast cancer is one of the most common cancers affecting the morbidity and mortality of Iranian women. This disease is a result of collective alterations of oncogenes and tumor suppressor genes. Studies have produced conflicting results concerning the role of p53 codon 72 polymorphism (G>C) and miR-146a rs2910164 polymorphism (G>C) on the risk of several cancers; therefore, a research was performed to estimate the association between the p53 codon 72 polymorphism and miR-146a rs2910164 polymorphism in breast cancer. Methods and Materials: A total of 45 archival breast cancer samples from khatam hospital and 40 healthy samples were collected. Verification of each cancer reported in a relative was sought through the pathology reports of the hospital records. Then, DNA extracted from all samples by standard methods and p53 codon 72 polymorphism genotypes and miR-146a rs2910164 polymorphism genotypes were analyzed using multiplex PCR. The tubules, mitotic activity, necrosis, polymorphism and grade of breast cancer were staged by Nottingham histological grading and immunohistochemical staining of the sections from the paraffin wax embedded tissues for the expression of ER, PR and p53 was carried out using a standard method. Finally, data analysis was performed using the 7 version of the Epi Info(TM) 2012 software and test chi-square(x2) for trend. Results: Successful DNA extraction was assessed by PCR amplification of b-actin gene (99 bp). According to the results, p53 GG genotype and miR-146a rs2910164 CC genotype was significantly associated with increased risk of breast cancer in the study population. In this study, we established that tumors of p53 GG genotype and miR-146a rs2910164 CC genotype exhibited higher mitotic activity, higher polymorphism, lower necrosis, lower tubules, higher ER- and PR-negatives and lower TP53-positives than the other genotypes. Conclusion: The present study provided preliminary evidence that a p53 GG genotype may effect breast cancer risk in the study population, interacting synergistically with miR-146a rs2910164 CC genotype. Our results demonstrate that the testing of p53 codon 72 polymorphism genotypes and miR-146a rs2910164 polymorphism genotypes in combination with clinical parameters can serve as major risk factors in the early identification of breast cancers.

Keywords: breast cancer, p53 codon 72 polymorphism, miR-146a rs2910164 polymorphism, genotypes

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1624 The Impact of Technology on Handicapped and Disability

Authors: George Kamil Kamal Abdelnor

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Every major educational institution has incorporated diversity, equity, and inclusion (DEI) principles into its administrative, hiring, and pedagogical practices. Yet these DEI principles rarely incorporate explicit language or critical thinking about disability. Despite the fact that according to the World Health Organization, one in five people worldwide is disabled, making disabled people the larger minority group in the world, disability remains the neglected stepchild of DEI. Drawing on disability studies and crip theory frameworks, the underlying causes of this exclusion of disability from DEI, such as stigma, shame, invisible disabilities, institutionalization/segregation/delineation from family, and competing models and definitions of disability are examined. This paper explores both the ideological and practical shifts necessary to include disability in university DEI initiatives. It offers positive examples as well as conceptual frameworks such as 'divers ability' for so doing. Using Georgetown University’s 2020-2022 DEI initiatives as a case study, this paper describes how curricular infusion, accessibility, identity, community, and diversity administration infused one university’s DEI initiatives with concrete disability-inclusive measures. It concludes with a consideration of how the very framework of DEI itself might be challenged and transformed if disability were to be included.

Keywords: cognitive disability, cognitive diversity, disability, higher education disability, Standardized Index of Diversity of Disability (SIDD), differential and diversity in disability, 60+ population diversity, equity, inclusion, crip theory, accessibility

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1623 Status and Image of the Nurse as Perceived by the Public

Authors: Salam Hadid, Mohammad Khatib

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The International Council of Nurses-ICN defined nursing as a sphere integrating autonomous and collaborative care intended for the individual, family and community within and outside of the care setting. Nursing as a care profession has developed broadly over recent decades in terms of its essentials, expertise and primarily academically. Despite the impressive growth of the profession, there is still extreme diversity in the public’s perceptions and opinions of the profession and its professionals and in the knowledge on the fundamentals of its true function and spheres of engagement. The current study examines the existing knowledge among the general population regarding the nursing profession. The population consisted of 498 respondents, 236 women and 262 men, age 18-81. The respondents noted that nursing focuses on the technical, and the emotional aspects and promotion of health for the patient are not the nurse’s responsibility. Most of the respondents saw nurses working mainly in hospital and community-based clinic settings. They considered nursing to be a high prestige profession in general, but less prestigious among respondents exposed to healthcare provision. Most of the respondents considered nursing to be a humane profession but without independence and with no need for academic studies. The findings are incompatible with the definition of nursing and its spheres of action as defined in the ICN Code of Ethics. Two suggestions are to work through nursing schools addressing the student nurses, as ambassadors for the profession. The second is using the healthcare encounter between the nursing staff and the public to improve the image of nurses.

Keywords: ethics, nurse image, public, nursing

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1622 Nurse’s Role in Early Detection of Breast Cancer through Mammography and Genetic Screening and Its Impact on Patient's Outcome

Authors: Salwa Hagag Abdelaziz, Dorria Salem, Hoda Zaki, Suzan Atteya

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Early detection of breast cancer saves many thousands of lives each year via application of mammography and genetic screening and many more lives could be saved if nurses are involved in breast care screening practices. So, the aim of the study was to identify nurse's role in early detection of breast cancer through mammography and genetic screening and its impact on patient's outcome. In order to achieve this aim, 400 women above 40 years, asymptomatic were recruited for mammography and genetic screening. In addition, 50 nurses and 6 technologists were involved in the study. A descriptive analytical design was used. Five tools were utilized: sociodemographic, mammographic examination and risk factors, women's before, during and after mammography, items relaying to technologists, and items related to nurses were also obtained. The study finding revealed that 3% of women detected for malignancy and 7.25% for fibroadenoma. Statistically, significant differences were found between mammography results and age, family history, genetic screening, exposure to smoke, and using contraceptive pills. Nurses have insufficient knowledge about screening tests. Based on these findings the present study recommended involvement of nurses in breast care which is very important to in force population about screening practices.

Keywords: mammography, early detection, genetic screening, breast cancer

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1621 Performance of an Improved Fluidized System for Processing Green Tea

Authors: Nickson Kipng’etich Lang’at, Thomas Thoruwa, John Abraham, John Wanyoko

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Green tea is made from the top two leaves and buds of a shrub, Camellia sinensis, of the family Theaceae and the order Theales. The green tea leaves are picked and immediately sent to be dried or steamed to prevent fermentation. Fluid bed drying technique is a common drying method used in drying green tea because of its ease in design and construction and fluidization of fine tea particles. Major problems in this method are significant loss of chemical content of the leaf and green appearance of tea, retention of high moisture content in the leaves and bed channeling and defluidization. The energy associated with the drying technology has been shown to be a vital factor in determining the quality of green tea. As part of the implementation, prototype dryer was built that facilitated sequence of operations involving steaming, cooling, pre-drying and final drying. The major findings of the project were in terms of quality characteristics of tea leaves and energy consumption during processing. The optimal design achieved a moisture content of 4.2 ± 0.84%. With the optimum drying temperature of 100 ºC, the specific energy consumption was 1697.8 kj.Kg-1 and evaporation rate of 4.272 x 10-4 Kg.m-2.s-1. The energy consumption in a fluidized system can be further reduced by focusing on energy saving designs.

Keywords: evaporation rate, fluid bed dryer, maceration, specific energy consumption

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1620 Socio-Economic Factors Influencing the Use of Coping Strategies among Conflict Actors (Farmers and Herders) in Giron Masa Village, Kebbi State, Nigeria

Authors: S. Umar, B. F. Umar

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This study was conducted at Giron Masa village, located 30 km from Yauri town. The study determines the socio-economic factors influencing the use of coping strategies among farmers and herders during post-conflict situation. Simple random sampling was employed to select one hundred respondents (50 farmers and 50 herders) from the study area. Logistic regression analysis (LR) was used to ascertain the socioeconomic variables that influenced the use of the coping strategies. The results of the study shows that age, income, family size and farming experience were individually significant and thus influenced the use of POCS by farmers. Annual income and production system influenced the use of POCS by herders. Age, farm size and farming experience were found to be individually significant in influencing the use of EOCS among farmers. Specifically, years of occupation experience among the herders increased the use of emotion oriented coping strategies among herders. The use of SSCS among farmers was influenced by educational level; farm size and farming experience, while the variables are not collectively significant in influencing the use of SSCS among the herders. The research recommends a need to adopt the strategy of community coping to cope with stress.

Keywords: farmers, herders, conflict, coping strategies

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1619 Effect of Ultrasound on Carotenoids Extraction from Pepper and Process Optimization Using Response Surface Methodology (RSM)

Authors: Elham Mahdian, Reza Karazhian, Rahele Dehghan Tanha

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Pepper (Capsicum annum L.) which belong to the family Solananceae, are known for their versatility as a vegetable crop and are consumed both as fresh vegetables or dehydrated for spices. Pepper is considered an excellent source of bioactive nutrients. Ascorbic acid, carotenoids and phenolic compounds are its main antioxidant constituents. Ultrasound assisted extraction is an inexpensive, simple and efficient alternative to conventional extraction techniques. The mechanism of action for ultrasound-assisted extraction are attributed to cavitations, mechanical forces and thermal impact, which result in disruption of cells walls, reduce particle size, and enhance mass transfer across cell membranes. In this study, response surface methodology was used to optimize experimental conditions for ultrasonic assisted extraction of carotenoid compounds from Chili peppers. Variables were included extraction temperatures at 3 levels (30, 40 and 50 °C), extraction times at 3 levels (10, 25 and 40 minutes) and power at 3 levels (30, 60 and 90 %). It was observed that ultrasound waves applied at temperature of 49°C, time of 10 minutes and power 89 % resulted to the highest carotenoids contents (lycopene and β-carotene), while the lowest value was recorded in the control. Thus, results showed that ultrasound waves have strong impact on extraction of carotenoids from pepper.

Keywords: carotenoids, optimization, pepper, response surface methodology

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1618 Stories of Digital Technology and Online Safety: Storytelling as a Tool to Find out Young Children’s Views on Digital Technology and Online Safety

Authors: Lindsey Watson

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This research is aimed at facilitating and listening to the voices of younger children, recognising their contributions to research about the things that matter to them. Digital technology increasingly impacts on the lives of young children, therefore this study aimed at increasing children’s agency through recognising and involving their perspectives to help contribute to a wider understanding of younger children’s perceptions of online safety. Using a phenomenological approach, the paper discusses how storytelling as a creative methodological approach enabled an agentic space for children to express their views, knowledge, and perceptions of their engagement with the digital world. Setting and parental informed consent were gained in addition to an adapted approach to child assent through the use of child-friendly language and emoji stickers, which was also recorded verbally. Findings demonstrate that younger children are thinking about many aspects of digital technology and how this impacts on their lives and that storytelling as a research method is a useful tool to facilitate conversations with young children. The paper thus seeks to recognise and evaluate how creative methodologies can provide insights into children’s understanding of online safety and how this can influence practitioners and parents in supporting younger children in a digital world.

Keywords: early childhood, family, online safety, phenomenology, storytelling

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1617 Epigenetic Modifying Potential of Dietary Spices: Link to Cure Complex Diseases

Authors: Jeena Gupta

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In the today’s world of pharmaceutical products, one should not forget the healing properties of inexpensive food materials especially spices. They are known to possess hidden pharmaceutical ingredients, imparting them the qualities of being anti-microbial, anti-oxidant, anti-inflammatory and anti-carcinogenic. Further aberrant epigenetic regulatory mechanisms like DNA methylation, histone modifications or altered microRNA expression patterns, which regulates gene expression without changing DNA sequence, contribute significantly in the development of various diseases. Changing lifestyles and diets exert their effect by influencing these epigenetic mechanisms which are thus the target of dietary phytochemicals. Bioactive components of plants have been in use since ages but their potential to reverse epigenetic alterations and prevention against diseases is yet to be explored. Spices being rich repositories of many bioactive constituents are responsible for providing them unique aroma and taste. Some spices like curcuma and garlic have been well evaluated for their epigenetic regulatory potential, but for others, it is largely unknown. We have evaluated the biological activity of phyto-active components of Fennel, Cardamom and Fenugreek by in silico molecular modeling, in vitro and in vivo studies. Ligand-based similarity studies were conducted to identify structurally similar compounds to understand their biological phenomenon. The database searching has been done by using Fenchone from fennel, Sabinene from cardamom and protodioscin from fenugreek as a query molecule in the different small molecule databases. Moreover, the results of the database searching exhibited that these compounds are having potential binding with the different targets found in the Protein Data Bank. Further in addition to being epigenetic modifiers, in vitro study had demonstrated the antimicrobial, antifungal, antioxidant and cytotoxicity protective effects of Fenchone, Sabinene and Protodioscin. To best of our knowledge, such type of studies facilitate the target fishing as well as making the roadmap in drug design and discovery process for identification of novel therapeutics.

Keywords: epigenetics, spices, phytochemicals, fenchone

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1616 Effect of Humic Substance on Ex-Vitro Propagation of Saffron (Crocus Sativus L.)

Authors: Abdelghani Tahiri, Youssef Karra, Naima Ait Aabd, Abdelaziz Mimouni

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Saffron (Crocus sativus L.), the most expensive spice in the world derived from the stigmas, is an autumn-flowering and sterile triploid (2n=3x=24) geophyte species that belong to the Iridaceae family. This plant species is mainly propagated vegetatively through the formation of daughter corms from the mother one. Low multiplication rates of daughter corms under natural conditions, along with fungal contamination, significantly reduce the productivity and quality of saffron corms. The development of efficient and sustainable strategies for rapid and large-scale production of selected cultivars of saffron will be desired. For this, the main objective of this work is to improve the vegetative propagation of saffron under ex-vitro conditions. Preliminary results of the influence of increasing doses of humic substances (HS) on the growth and multiplication of corms under greenhouse conditions are evaluated. The obtained data shows that the effect of HS depends on the concentration used and the mode of application. Indeed, the application through irrigation has increased the number of shoots and corms, but it has reduced other parameters. On the other hand, the temporary treatment has improved all observed parameters except for the number of shoots and corms. Results obtained in this work suggest that it is possible to improve the propagation of saffron corms under greenhouse conditions.

Keywords: saffron, Crocus sativus L., corm, humic substances

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1615 An Interrogation of Lecturer’s Skills in Assisting Visually Impaired Students during the COVID-19 Lockdown Era in Selected Universities in Zimbabwe

Authors: Esther Mafunda

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The present study interrogated the lecturer’s skills in supporting visually impaired students during the Covid-19 era at the University of Zimbabwe. It particularly assesses how the Covid-19 pandemic affected the learning experience of visually impaired students and which skills the lecturers possessed in order to assist the visually impaired students during online learning. Data was collected from lecturers and visually impaired students at the University of Zimbabwe Disability Resource Centre. Data was collected through the use of interviews and questionnaires. Using content analysis, it was established that visually impaired students faced challenges of lack of familiarity with the Moodle learning platform, marginalization, lack of professional training, and lack of training for parents and guardians. Lecturers faced challenges of lack of training, the curriculum, access, and technical know-how deficit. It was established that lecturers had to resort to social media platforms in order to assist visually impaired students. Visually impaired students also received assistance from their friends and family members. On the basis of the results of the research, it can be concluded that lecturers needed in-service training to be provided with the necessary skills and knowledge to teach students with visual impairments and provide quality education to students with visual impairments.

Keywords: visual impairment, disability, covid-19, inclusive learning

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1614 MicroRNA in Bovine Corpus Luteum during Early Pregnancy

Authors: Rreze Gecaj, Corina Schanzenbach, Benedikt Kirchner, Michael Pfaffl, Bajram Berisha

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The maintenance of corpus lutem (CL) during early pregnancy in cattle is a critical and multifarious process. A luteotrophic mechanism originating from the embryo is widely accepted as the triggering signal for the CL maintenance. In the cattle, it is the interferon-tau (IFNT) secretion form conceptus that prevents CL regression and ensures progesterone production for the establishment of pregnancy. In addition to endocrine and paracrine signals, microRNA (miRNA) can also support CL sustainability during early pregnancy. MiRNA are small non-coding nucleic acids that regulate gene expression post-transcriptionally and are shown to be involved in the modulation of CL function. However, the examination of miRNAs in corpus luteum function at the early pregnancy still remains largely uncovered. This study aims at profiling the expression of miRNA in CL during the early pregnancy in cattle by comparing it with the CL form late cycle and with the regressed CL. Corpora lutea were assigned in two different groups during the cycle (C13 group, late CL: days 13-18 and C18, regressed CL group: day >18) and during the early pregnancy (group P: 1-2 month). The estrous cycle was determined by macroscopic examination and to age the fetus crown-rump length measurement was applied. A total of 9 corpora lutea from individual animals were included in the study, three corpora lutea for each group. MiRNAs population was profiled using small RNA next-generation sequencing and biologically significant miRNAs were evaluated for their differential expression using the DESeq2-methodology. We show that 6 differentially expressed miRNAs (bta-mir-2890, -2332, -2441-3p, -148b, -1248 and -29c) are common to both comparisons, P vs C13 and P vs C18. While for each stage individually we have identified unique miRNAs differentially expressed only for the given comparison. bta-miR-23a and -769 were unique miRNAs differentially expressed in P vs C13, whereas forty-four unique miRNAs were identified as differentially expressed in P vs C18. These data confirm that miRNAs are highly abundant in luteal tissue during early pregnancy and potentially regulate the CL maintenance at this stage of fetus development.

Keywords: bovine, corpus luteum, microRNA, pregnancy, RNA-Seq

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1613 Dietary Nutrient Consumption Patterns by the Pregnant Mother in Dhaka City, Bangladesh

Authors: Kazi Muhammad Rezaul Karim, Tasmia Tasnim

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Introduction: Pregnancy is a condition of higher nutrient requirement but in developing countries like Bangladesh most of the pregnant women can not meet their nutrient requirement and sometimes they are neglected in the family. The purpose of the study was to assess the nutritional status and dietary nutrient intake by the pregnant women, in Dhaka city, Bangladesh. Methods: The study population comprised of pregnant women from urban or semi-urban, aged between 18 to 35 and free of pregnancy related complication and other diseases. Under a cross-sectional design, 30 healthy non-pregnant as well as 130 pregnant women, at 3 different trimesters of pregnancy were assessed. A questionnaire was developed to obtain demographic, socio-economic, anthropometric, drug and medical history. Three day consecutive 24-hour food recalls were used to assess food intake and then converted to nutrient intake. Results: The average BMI of the nonpregnant women was 22.89 ± 3.4 kg/m2 and that of pregnant women was 23.52 ± 3.71 kg/m2. The mean dietary nutrient intake of dietary fiber, calorie, protein, fat, carbohydrate, calcium, iron, thiamine, riboflavin, vitamin C, Vitamin A, folate, vitamin B6 and Vitamin B12 of the pregnant mothers were 4.38 g, 1619 kcal, 60.05 g, 30.38 g, 268.79 g, 537.21 mg, 21.53 mg, 1.15 mg, 0.94 mg, 97.36 mg, 647.6 µg, 153.93 µg, 1.41 mg and 4.09 µg respectively. Most of pregnant women (more than 90%) can not meet their energy, calcium and folate requirements. Conclusion: Most of the pregnant mother in Bangladesh can not meet their dietary requirements during pregnancy.

Keywords: pregnancy, dietary nutrient, nutritional status, BMI

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1612 Effect of Mistranslating tRNA Alanine on Polyglutamine Aggregation

Authors: Sunidhi Syal, Rasangi Tennakoon, Patrick O'Donoghue

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Polyglutamine (polyQ) diseases are a group of diseases related to neurodegeneration caused by repeats of the amino acid glutamine (Q) in the DNA, which translates into an elongated polyQ tract in the protein. The pathological explanation is that the polyQ tract forms cytotoxic aggregates in the neurons, leading to their degeneration. There are no cures or preventative efforts established for these diseases as of today, although the symptoms of these diseases can be relieved. This study specifically focuses on Huntington's disease, which is a type of polyQ disease in which aggregation is caused by the extended cytosine, adenine, guanine (CUG) codon repeats in the huntingtin (HTT) gene, which encodes for the huntingtin protein. Using this principle, we attempted to create six models, which included mutating wildtype tRNA alanine variant tRNA-AGC-8-1 to have glutamine anticodons CUG and UUG so serine is incorporated at glutamine sites in poly Q tracts. In the process, we were successful in obtaining tAla-8-1 CUG mutant clones in the HTTexon1 plasmids with a polyQ tract of 23Q (non-pathogenic model) and 74Q (disease model). These plasmids were transfected into mouse neuroblastoma cells to characterize protein synthesis and aggregation in normal and mistranslating cells and to investigate the effects of glutamines replaced with alanines on the disease phenotype. Notably, we observed no noteworthy differences in mean fluorescence between the CUG mutants for 23Q or 74Q; however, the Triton X-100 assay revealed a significant reduction in insoluble 74Q aggregates. We were unable to create a tAla-8-1 UUG mutant clone, and determining the difference in the effects of the two glutamine anticodons may enrich our understanding of the disease phenotype. In conclusion, by generating structural disruption with the amino acid alanine, it may be possible to find ways to minimize the toxicity of Huntington's disease caused by these polyQ aggregates. Further research is needed to advance knowledge in this field by identifying the cellular and biochemical impact of specific tRNA variants found naturally in human genomes.

Keywords: Huntington's disease, polyQ, tRNA, anticodon, clone, overlap PCR

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1611 Characterization of Penicillin V Acid and Its Related Compounds by HPLC

Authors: Bahdja Guerfi, N. Hadhoum, I. Azouz, M. Bendoumia, S. Bouafia, F. Z. Hadjadj Aoul

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Background: 'Penicillin V' is a narrow, bactericidal antibiotic of the beta-lactam family of the naturally occurring penicillin group. It is limited to infections due to the germs defined as sensitive. The objective of this work was to identify and to characterize Penicillin V acid and its related compounds by High-performance liquid chromatography (HPLC). Methods: Firstly phenoxymethylpenicillin was identified by an infrared absorption. The organoleptic characteristics, pH, and determination of water content were also studied. The dosage of Penicillin V acid active substance and the determination of its related compounds were carried on waters HPLC, equipped with a UV detector at 254 nm and Discovery HS C18 column (250 mm X 4.6 mm X 5 µm) which is maintained at room temperature. The flow rate was about 1 ml per min. A mixture of water, acetonitrile and acetic acid (65:35:01) was used as mobile phase for phenoxyacetic acid ‘impurity B' and a mixture of water, acetonitrile and acetic acid (650:150:5.75) for the assay and 4-hydroxypenicillin V 'impurity D'. Results: The identification of Penicillin V acid active substance and the evaluation of its chemical quality showed conformity with USP 35th edition. The Penicillin V acid content in the raw material is equal to 1692.22 UI/mg. The percentage content of phenoxyacetic acid and 4-hydroxypenicillin V was respectively: 0.035% and 0.323%. Conclusion: Through these results, we can conclude that the Penicillin V acid active substance tested is of good physicochemical quality.

Keywords: characterization, HPLC, Penicillin V acid, related substances

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1610 Mental Health Literacy in the Arabic Community

Authors: Yamam Abuzinadah

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Mental health literacy has become a very influential topic around the world due to the increase of mental health issues that have been reported through national research and surveys. Mental health literacy refers to the awareness, attitudes, beliefs, knowledge and skills when dealing with mental illness. This research explores mental health literacy in the Arabic and the ways culture informs perceptions of mental health in general. Also, the impact of mental health literacy on: help-seeking attitudes, relationships and community interactions. The outcomes of this research will contribute to raising mental health awareness among the Arabic community, develop and enhance mental health service provision and explore new ideas in regards to elevating mental health literacy in the Arabic community. This research aims to explore attitudes, beliefs, perspective, values and perceptions toward mental health in general among the Arabic community. It will also aim to highlight the factors contributing to theses beliefs, perspective, value and perception and accordingly the role these factors play in regards to awareness, services access, recovery and care provided from the family and the community. This thesis will aim to reflect a detailed theorisation and exploration of: (1) The impact of cultural factors on mental health literacy ie. attitudes, beliefs, knowledge and skills. (2) The ways culture informs perceptions of mental health literacy. (3) The impact of mental health literacy on: help-seeking behaviors, and relationships and community interactions.

Keywords: Arab, mental health, literacy, awareness

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1609 Hepatocyte-Intrinsic NF-κB Signaling Is Essential to Control a Systemic Viral Infection

Authors: Sukumar Namineni, Tracy O'Connor, Ulrich Kalinke, Percy Knolle, Mathias Heikenwaelder

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The liver is one of the pivotal organs in vertebrate animals, serving a multitude of functions such as metabolism, detoxification and protein synthesis and including a predominant role in innate immunity. The innate immune mechanisms pertaining to liver in controlling viral infections have largely been attributed to the Kupffer cells, the locally resident macrophages. However, all the cells of liver are equipped with innate immune functions including, in particular, the hepatocytes. Hence, our aim in this study was to elucidate the innate immune contribution of hepatocytes in viral clearance using mice lacking Ikkβ specifically in the hepatocytes, termed IkkβΔᴴᵉᵖ mice. Blockade of Ikkβ activation in IkkβΔᴴᵉᵖ mice affects the downstream signaling of canonical NF-κB signaling by preventing the nuclear translocation of NF-κB, an important step required for the initiation of innate immune responses. Interestingly, infection of IkkβΔᴴᵉᵖ mice with lymphocytic choriomeningitis virus (LCMV) led to strongly increased hepatic viral titers – mainly confined in clusters of infected hepatocytes. This was due to reduced interferon stimulated gene (ISG) expression during the onset of infection and a reduced CD8+ T-cell-mediated response. Decreased ISG production correlated with increased liver LCMV protein and LCMV in isolated hepatocytes from IkkβΔᴴᵉᵖ mice. A similar phenotype was found in LCMV-infected mice lacking interferon signaling in hepatocytes (IFNARΔᴴᵉᵖ) suggesting a link between NFkB and interferon signaling in hepatocytes. We also observed a failure of interferon-mediated inhibition of HBV replication in HepaRG cells treated with NF-kB inhibitors corroborating our initial findings with LCMV infections. Collectively, these results clearly highlight a previously unknown and influential role of hepatocytes in the induction of innate immune responses leading to viral clearance during a systemic viral infection with LCMV-WE.

Keywords: CD8+ T cell responses, innate immune mechanisms in the liver, interferon signaling, interferon stimulated genes, NF-kB signaling, viral clearance

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1608 Rare Case of Pyoderma Gangrenosum of the Upper Limb

Authors: Karissa A. Graham

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Pyoderma gangrenosum (PG) is a prototypic autoinflammatory neutrophilic dermatosis that is a rare disorder. It presents a diagnostic challenge owing to its variable presentation, clinical overlap with other conditions, it is often associated with other systemic conditions, and there is no definitive histological or laboratory characteristic. The Delphai consensus for PG includes the presence of at least one ulcer on the anterior lower limb. Systemic corticosteroids and immunosuppressive therapies are the mainstay treatment for PG. We describe a case report of delayed diagnosis of ulcerative pyoderma gangrenosum in a 44-year-old male on his forearm. The patient presented with an infected ulcer on his right forearm that had been present for over three years. The patient was a Type 2 Diabetic with no personal or family history of inflammatory bowel disease or other autoimmune diseases. The patient was initially investigated for malignancy, but biopsies returned as chronic inflammatory tissue with neutrophilic infiltrate and no malignancy. The patient was commenced on systemic prednisone for the treatment of pyoderma gangrenosum. The diagnosis of ulcerative PG poses a challenge given the vast differential diagnosis for a cutaneous ulcer (i.e., malignant, vascular, autoimmune, trauma, infective, etc.). Diagnostic accuracy is important given that the treatment for PG with steroids does not go without risks and indeed may be contraindicated in other potential causes of the ulcer. Indeed, more common and more sinister causes of ulcers should be investigated first, as death from PG is quite rare.

Keywords: dermatological diagnosis, dermatosis, pyoderma gangrenosum, rare presentation

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1607 The Comparison Study of Human Microbiome in Chronic Rhinosinusitis between Adults and Children

Authors: Il Ho Park, Joong Seob Lee, Sung Hun Kang, Jae-Min Shin, Il Seok Park, Seok Min Hong, Seok Jin Hong

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Introduction: The human microbiota is the aggregate of microorganisms, and the bacterial microbiome of the human digestive tract contributes to both health and disease. In health, bacteria are key components in the development of mucosal barrier function and in innate and adaptive immune responses, and they also work to suppress the establishment of pathogens. In human upper airway, the sinonasal microbiota might play an important role in chronic rhinosinusitis (CRS). The purpose of this study is to investigate the human upper airway microbiome in CRS patients and to compare the sinonasal microbiome of adults with children. Materials and methods: A total of 19 samples from 19 patients (Group1; 9 CRS in children, aged 5 to 14 years versus Group 2; 10 CRS in adults aged 21 to 59 years) were examined. Swabs were collected from the middle meatus and/or anterior ethmoid region under general anesthesia during endoscopic sinus surgery or tonsillectomy. After DNA extraction from swab samples, we analysed bacterial microbiome consortia using 16s rRNA gene sequencing approach (the Illumina MiSeq platform). Results: In this study, relatively abundance of the six bacterial phyla and tremendous genus and species found in substantial amounts in the individual sinus swab samples, include Corynebacterium, Hemophilus, Moraxella, and Streptococcus species. Anaerobes like Fusobacterium and Bacteroides were abundantly present in the children group, Bacteroides and Propionibacterium were present in adults group. In genus, Haemophilus was the most common CRS microbiome in children and Corynebacterium was the most common CRS microbiome in adults. Conclusions: Our results show the diversity of human upper airway microbiome, and the findings will suggest that CRS is a polymicrobial infection. The Corynebacterium and Hemophilus may live as commensals on mucosal surfaces of sinus in the upper respiratory tract. The further study will be needed for analysis of microbiome-human interactions in upper airway and CRS.

Keywords: microbiome, upper airway, chronic rhinosinusitis, adult and children

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1606 In Silico Study of Cell Surface Structures of Parabacteroides distasonis Involved in Its Maintain Within the Gut Microbiota and Its Potential Pathogenicity

Authors: Jordan Chamarande, Lisiane Cunat, Corentine Alauzet, Catherine Cailliez-Grimal

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Gut microbiota (GM) is now considered a new organ mainly due to the microorganism’s specific biochemical interaction with its host. Although mechanisms underlying host-microbiota interactions are not fully described, it is now well-defined that cell surface molecules and structures of the GM play a key role in such relation. The study of surface structures of GM members is also fundamental for their role in the establishment of species in the versatile and competitive environment of the digestive tract and as a potential virulence factor. Among these structures are capsular polysaccharides (CPS), fimbriae, pili and lipopolysaccharides (LPS), all well-described for their central role in microorganism colonization and communication with host epithelium. The health-promoting Parabacteroides distasonis, which is part of the core microbiome, has recently received a lot of attention, showing beneficial properties for its host and as a new potential biotherapeutic product. However, to the best of the authors’ knowledge, the cell surface molecules and structures of P. distasonis that allow its maintain within the GM are not identified. Moreover, although P. distasonis is strongly recognized as intestinal commensal species with benefits for its host, it has also been recognized as an opportunistic pathogen. In this study, we reported gene clusters potentially involved in the synthesis of the capsule, fimbriae-like and pili-like cell surface structures in 26 P. distasonis genomes and applied the new RfbA-Typing classification in order to better understand and characterize the beneficial/pathogenic behaviour related to P. distasonis strains. In context, 2 different types of fimbriae, 3 of pilus and up to 14 capsular polysaccharide loci, have been identified over the 26 genomes studied. Moreover, the addition of data to the rfbA-Type classification modified the outcome by rearranging rfbA genes and adding a fifth group to the classification. In conclusion, the strain variability in terms of external proteinaceous structure could explain the inter-strain differences previously observed in P. distasonis adhesion capacities and its potential pathogenicity.

Keywords: gut microbiota, Parabacteroides distasonis, capsular polysaccharide, fimbriae, pilus, O-antigen, pathogenicity, probiotic, comparative genomics

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1605 Impact of Diet and COVID-19 Policies on Osteopenia in a Hispanic White Adolescent Girl

Authors: Maria Angelica Trak-Fellermeier, Alison K. Macchi, Rodolfo Galvan, Yolangel Hernandez, Thresia Gambon, Rebeca Martinez, Cristina Palacios

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Poor lifestyle habits, vitamin D deficiency, and inadequate calcium intake, particularly during the COVID-19 pandemic, may contribute to severe osteopenia in childhood, increasing future fractures and osteoporosis risk. We here present a case of osteopenia in a 13-year-old white, Hispanic, premenarchal girl who completed the baseline visit of the MetA-Bone Trial during the COVID-19 pandemic. The premenarchal girl has a family history of osteoporosis (maternal grandfather) but no previous fractures; moderate outdoor activity was <1 hour/day 3 times/week with 8 hours/day of sleep. Consumption of dairy products and vegetables was <1 serving/day. Lab blood tests confirmed vitamin D deficiency (serum 25(OH)D: 9 ng/L) and hyperphosphatemia (5.2 mg/dL); other tests were normal. DXA scan Z‐score was ‐2.2 SD (indicative of osteopenia by age and sex). The premenarchal girl was referred to a pediatrician, who confirmed the results, and prescribed a daily supplement with 2000 IU of vitamin D and 1000 mg of calcium. Seclusion during the COVID-19 pandemic may have contributed to the severity of the findings. Therefore, we recommend screening children undergoing growth spurts for vitamin D, calcium, and poor lifestyle habits during and after the pandemic.

Keywords: bone mass, vitamin D, puberty, Hispanic

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1604 Micromechanical Compatibility Between Cells and Scaffold Mediates the Efficacy of Regenerative Medicine

Authors: Li Yang, Yang Song, Martin Y. M. Chiang

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Objective: To experimentally substantiate the micromechanical compatibility between cell and scaffold, in the regenerative medicine approach for restoring bone volume, is essential for phenotypic transitions Methods: Through nanotechnology and electrospinning process, nanofibrous scaffolds were fabricated to host dental follicle stem cells (DFSCs). Blends (50:50) of polycaprolactone (PCL) and silk fibroin (SF), mixed with various content of cellulose nanocrystals (CNC, up to 5% in weight), were electrospun to prepare nanofibrous scaffolds with heterogeneous microstructure in terms of fiber size. Colloidal probe atomic force microscopy (AFM) and conventional uniaxial tensile tests measured the scaffold stiffness at the micro-and macro-scale, respectively. The cell elastic modulus and cell-scaffold adhesive interaction (i.e., a chemical function) were examined through single-cell force spectroscopy using AFM. The quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was used to determine if the mechanotransduction signal (i.e., Yap1, Wwr2, Rac1, MAPK8, Ptk2 and Wnt5a) is upregulated by the scaffold stiffness at the micro-scale (cellular scale). Results: The presence of CNC produces fibrous scaffolds with a bimodal distribution of fiber diameter. This structural heterogeneity, which is CNC-composition dependent, remarkably modulates the mechanical functionality of scaffolds at microscale and macroscale simultaneously, but not the chemical functionality (i.e., only a single material property is varied). In in vitro tests, the osteogenic differentiation and gene expression associated with mechano-sensitive cell markers correlate to the degree of micromechanical compatibility between DFSCs and the scaffold. Conclusion: Cells require compliant scaffolds to encourage energetically favorable interactions for mechanotransduction, which are converted into changes in cellular biochemistry to direct the phenotypic evolution. The micromechanical compatibility is indeed important to the efficacy of regenerative medicine.

Keywords: phenotype transition, scaffold stiffness, electrospinning, cellulose nanocrystals, single-cell force spectroscopy

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1603 “Divorced Women are Like Second-Hand Clothes” - Hate Language in Media Discourse (Using the Example of Electronic Media Platforms)

Authors: Sopio Totibadze

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Although the legal framework of Georgia reflects the main principles of gender equality and is in line with the international situation (UNDP, 2018), Georgia remains a male-dominated society. This means that men prevail in many areas of social, economic, and political life, which frequently gives women a subordinate status in society and the family (UN women). According to the latest study, “violence against women and girls in Georgia is also recognized as a public problem, and it is necessary to focus on it” (UN women). Moreover, the Public Defender's report on the protection of human rights in Georgia (2019) reveals that “in the last five years, 151 women were killed in Georgia due to gender and family violence”. Sadly, these statistics have increased significantly since that time. The issue was acutely reflected in the document published by the Organization for Security and Cooperation in Europe, “Gender Hate Crime” (March 10, 2021). “Unfortunately, the rates of femicide ..... are still high in the country, and distrust of law enforcement agencies often makes such cases invisible, which requires special attention from the state.” More precisely, the cited document considers that there are frequent cases of crimes based on gender-based oppression in Georgia, which pose a threat not only to women but also to people of any gender whose desires and aspirations do not correspond to the gender norms and roles prevailing in society. According to the study, this type of crime has a “significant and lasting impact on the victim(s) and also undermines the safety and cohesion of society and gender equality”. It is well-known that language is often used as a tool for gender oppression (Rusieshvili-Cartledge and Dolidze, 2021; Totibadze, 2021). Therefore, feminist and gender studies in linguistics ultimately serve to represent the problem, reflect on it, and propose ways to solve it. Together with technical advancement in communication, a new form of discrimination has arisen- hate language against women in electronic media discourse. Due to the nature of social media and the internet, messages containing hate language can spread in seconds and reach millions of people. However, only a few know about the detrimental effects they may have on the addressee and society. This paper aims to analyse the hateful comments directed at women on various media platforms to determine (1) the linguistic strategies used while attacking women and (2) the reasons why women may fall victim to this type of hate language. The data have been collected over six months, and overall, 500 comments will be examined for the paper. Qualitative and quantitative analysis was chosen for the methodology of the study. The comments posted on various media platforms, including social media posts, articles, or pictures, have been selected manually due to several reasons, the most important being the problem of identifying hate speech as it can disguise itself in different ways- humour, memes, etc. The comments on the articles, posts, pictures, and videos selected for sociolinguistic analysis depict a woman, a taboo topic, or a scandalous event centred on a woman that triggered a lot of hatred and hate language towards the person to whom the post/article was dedicated. The study has revealed that a woman can become a victim of hatred directed at them if they do something considered to be a deviation from a societal norm, namely, get a divorce, be sexually active, be vocal about feministic values, and talk about taboos. Interestingly, people who utilize hate language are not only men trying to “normalize” the prejudiced patriarchal values but also women who are equally active in bringing down a "strong" woman. The paper also aims to raise awareness about the hate language directed at women, as being knowledgeable about the issue at hand is the first step to tackling it.

Keywords: femicide, hate language, media discourse, sociolinguistics

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1602 Ultrasensitive Detection and Discrimination of Cancer-Related Single Nucleotide Polymorphisms Using Poly-Enzyme Polymer Bead Amplification

Authors: Lorico D. S. Lapitan Jr., Yihan Xu, Yuan Guo, Dejian Zhou

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The ability of ultrasensitive detection of specific genes and discrimination of single nucleotide polymorphisms is important for clinical diagnosis and biomedical research. Herein, we report the development of a new ultrasensitive approach for label-free DNA detection using magnetic nanoparticle (MNP) assisted rapid target capture/separation in combination with signal amplification using poly-enzyme tagged polymer nanobead. The sensor uses an MNP linked capture DNA and a biotin modified signal DNA to sandwich bind the target followed by ligation to provide high single-nucleotide polymorphism discrimination. Only the presence of a perfect match target DNA yields a covalent linkage between the capture and signal DNAs for subsequent conjugation of a neutravidin-modified horseradish peroxidase (HRP) enzyme through the strong biotin-nuetravidin interaction. This converts each captured DNA target into an HRP which can convert millions of copies of a non-fluorescent substrate (amplex red) to a highly fluorescent product (resorufin), for great signal amplification. The use of polymer nanobead each tagged with thousands of copies of HRPs as the signal amplifier greatly improves the signal amplification power, leading to greatly improved sensitivity. We show our biosensing approach can specifically detect an unlabeled DNA target down to 10 aM with a wide dynamic range of 5 orders of magnitude (from 0.001 fM to 100.0 fM). Furthermore, our approach has a high discrimination between a perfectly matched gene and its cancer-related single-base mismatch targets (SNPs): It can positively detect the perfect match DNA target even in the presence of 100 fold excess of co-existing SNPs. This sensing approach also works robustly in clinical relevant media (e.g. 10% human serum) and gives almost the same SNP discrimination ratio as that in clean buffers. Therefore, this ultrasensitive SNP biosensor appears to be well-suited for potential diagnostic applications of genetic diseases.

Keywords: DNA detection, polymer beads, signal amplification, single nucleotide polymorphisms

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1601 Expression of DNMT Enzymes-Regulated miRNAs Involving in Epigenetic Event of Tumor and Margin Tissues in Patients with Breast Cancer

Authors: Fatemeh Zeinali Sehrigh

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Background: Breast cancer is the most common and deadly cancer among women. The incidence of breast cancer depends on various genetic, epigenetic and environmental factors. DNA methylation and miRNAs are the most important epigenetic factors involved in breast cancer. miRNAs play an important role in the post-transcriptional regulation of genes, including genes involved in DNA methylation (DNMTs), and are also important regulators of oncogenic pathways. The study of microRNAs and DNMTs in Breast cancer allows the development of targeted treatments and early detection of this cancer. Methods and Materials: Experimental Statistical significance of differences in miRNAs and DNMTs relative expression results were analyzed by pair-wise fixed reallocation randomization test as a statistical model included in the REST (relative expression software tool). In silico analysis confirmed that microarray datasets miRNAs and DNMTs were downloaded from the GEO database. The analysis was defined using the Affy packages in R software to screen remarkably dysregulated miRNAs and DNMTs. the target genes in terms of molecular pathways in the web, Gestalt and GO (Gene Ontology) that specify the biological processes of molecular functions and cellular components to be investigated. In laboratory studies, sampling was done from 100 women with breast cancer and 100 healthy women with written consent. Tissue RNA was extracted using a Trizol solution. Then, using the cDNA synthesis kit, DNA was synthesized from the extracted RNAs, and the expression level of miR-509-3-5p and miR-596 was investigated using the Realtime PCR method. Results: In the present study, we found significant (p-value<0.05) changes in the expression level of miRNAs and DNMTs in patients with breast cancer. In bioinformatics studies, the GEO microarray data set, similar to qPCR results, showed a decreased expression of miRNAs and an increased expression of DNMTs in breast cancer. Conclusion: The results of the present study, which showed a decrease in the expression of miRNAs and DNMTs in breast cancer, suggest that these genes can be used as important diagnostic and therapeutic biomarkers in breast cancer.

Keywords: expression omnibus (GEO), microarray dataset, breast cancer, miRNA

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1600 Gender and Citizen Participation at the Local Governments: A Case of Vietnam

Authors: Trinh Hoang Hong Hue

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Citizen Participation has been largely considered as an important objective of improving democracy and government decision-making in Vietnam recently. The Public Administration Performance Index Survey data (PAPI) indicated that citizens in provinces that have a higher proportion of male often less participate in local governance than those in provinces that have lower proportion of male. That means Vietnamese women more actively participate at the local governance rather than men. Thus this study will explore factors involving gender differences that impact on citizen participation at the local level. Applying qualitative approach, mainly in-depth interview, this study explores four diverse perspectives on enhancing citizen participation for both women and men at the local governance including civic knowledge; the trust of citizens; suitable policies of local government; and the role of NGOs. Furthermore, this study also points out two crucial reasons that are leading to the gender differences of citizen participation at the local level. Firstly, because Vietnamese women play the main role in family financial management; then they are willing to highly contribute to ‘voluntary contributions’; one of the four sub-dimensions of the concept ‘citizen participation’ of PAPI. Secondly, in Vietnam, women are deeply prone to be interested in the small issues at the local governance; whereas men are much keen on the bigger issues at national and international governance.

Keywords: citizen participation, gender, women, local governance, PAPI, Vietnam

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1599 The Effect of a Probiotic Diet on htauE14 in a Rodent Model of Alzheimer’s Disease

Authors: C. Flynn, Q. Yuan, C. Reinhardt

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Alzheimer’s Disease (AD) is a progressive neurodegenerative disorder affecting broad areas of the cerebral cortex and hippocampus. More than 95% of AD cases are representative of sporadic AD, where both genetic and environmental risk factors play a role. The main pathological features of AD include the widespread deposition of amyloid-beta and neurofibrillary tau tangles in the brain. The earliest brain pathology related to AD has been defined as hyperphosphorylated soluble tau in the noradrenergic locus coeruleus (LC) neurons, characterized by Braak. However, the cause of this pathology and the ultimate progression of AD is not understood. Increasing research points to a connection between the gut microbiota and the brain, and mounting evidence has shown that there is a bidirectional interaction between the two, known as the gut-brain axis. This axis can allow for bidirectional movement of neuroinflammatory cytokines and pathogenic misfolded proteins, as seen in AD. Prebiotics and probiotics have been shown to have a beneficial effect on gut health and can strengthen the gut-barrier as well as the blood-brain barrier, preventing the spread of these pathogens across the gut-brain axis. Our laboratory has recently established a pretangle tau rat model, in which we selectively express pseudo-phosphorylated human tau (htauE14) in the LC neurons of TH-Cre rats. LC htauE14 produced pathological changes in rats resembling those of the preclinical AD pathology (reduced olfactory discrimination and LC degeneration). In this work, we will investigate the effects of pre/probiotic ingestion on AD behavioral deficits, blood inflammation/cytokines, and various brain markers in our experimental rat model of AD. Rats will be infused with an adeno-associated viral vector containing a human tau gene pseudophosphorylated at 14 sites (common in LC pretangles) into 2-3 month TH-Cre rats. Fecal and blood samples will be taken at pre-surgery, and various post-surgery time points. A collection of behavioral tests will be performed, and immunohistochemistry/western blotting techniques will be used to observe various biomarkers. This work aims to elucidate the relationship between gut health and AD progression by strengthening gut-brain relationship and aims to observe the overall effect on tau formation and tau pathology in AD brains.

Keywords: alzheimer’s disease, aging, gut microbiome, neurodegeneration

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1598 Molecular Diversity of Forensically Relevant Insects from the Cadavers of Lahore

Authors: Sundus Mona, Atif Adnan, Babar Ali, Fareeha Arshad, Allah Rakha

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Molecular diversity is the variation in the abundance of species. Forensic entomology is a neglected field in Pakistan. Insects collected from the crime scene should be handled by forensic entomologists who are currently virtually non-existent in Pakistan. Correct identification of insect specimen along with knowledge of their biodiversity can aid in solving many problems related to complicated forensic cases. Inadequate morphological identification and insufficient thermal biological studies limit the entomological utility in Forensic Medicine. Recently molecular identification of entomological evidence has gained attention globally. DNA barcoding is the latest and established method for species identification. Only proper identification can provide a precise estimation of postmortem intervals. Arthropods are known to be the first tourists scavenging on decomposing dead matter. The objective of the proposed study was to identify species by molecular techniques and analyze their phylogenetic importance with barcoded necrophagous insect species of early succession on human cadavers. Based upon this identification, the study outcomes will be the utilization of established DNA bar codes to identify carrion feeding insect species for concordant estimation of post mortem interval. A molecular identification method involving sequencing of a 658bp ‘barcode’ fragment of the mitochondrial cytochrome oxidase subunit 1 (CO1) gene from collected specimens of unknown dipteral species from cadavers of Lahore was evaluated. Nucleotide sequence divergences were calculated using MEGA 7 and Arlequin, and a neighbor-joining phylogenetic tree was generated. Three species were identified, Chrysomya megacephala, Chrysomya saffranea, and Chrysomya rufifacies with low genetic diversity. The fixation index was 0.83992 that suggests a need for further studies to identify and classify forensically relevant insects in Pakistan. There is an exigency demand for further research especially when immature forms of arthropods are recovered from the crime scene.

Keywords: molecular diversity, DNA barcoding, species identification, forensically relevant

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