Search results for: gene family
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 4414

Search results for: gene family

3154 Love Is Color-Blind: Perceptions among Sunway University Students toward Interracial Relationship

Authors: Azman Ramlie, Vivian Foo Jing Wen

Abstract:

As the advancement of technology has tremendously changed the way people communicate with each other, it has opened up opportunities for interracial relationships. Interracial relationship is also known as intercultural or interethnic relationships. This research topic aims to study on students’ perceptions toward interracial relationship in terms of their level of acceptance and approval. In addition, the study also focuses on determining and ranking the factors that affects students’ perceptions towards interracial relationship. This study also targets to determine the differences in students’ perceptions from business and communication major. This study uses a sample of university students from Sunway University, particularly on students in business and communication major. The study was conducted through online survey questionnaires. Results revealed that students’ level of acceptance towards interracial relationship was high. In addition, results also further indicated that family members’ support was one of the most important factors in influencing students’ perception towards interracial relationship. No diverse differences of perceptions among students in business and communication major towards interracial relationship were shown in the findings. Most of the result showed that both majors turned out to have similar perceptions towards interracial relationship. It can be said that the students’ perception towards interracial relationship did not differ from which major the students was in but their family background that would shape their perceptions.

Keywords: interracial relationship, racial, relationship, perceptions

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3153 Phylogenetic Relationships of Aproaerema Simplexella (Walker) and the Groundnut Leaf Miner Aproaerema Modicella (Deventer) (Lepidoptera: Gelechiidae) Collected from Australia, India, Mozambique, and South Africa

Authors: Makhosi Buthelezi

Abstract:

Mitochondrial DNA cytochrome c oxidase I (COI) gene analyses linked the South African groundnut leaf miner (GLM) to the Australian soya bean moth Aproaerema simplexella (Walker) and Indian Aproaerema modicella (Deventer). Thus, the genetic relatedness of GLM, A. simplexela, and A. modicella was examined by performing mitochondrial and nuclear (COI, cytochrome oxidase subunit II (COII), mitochondrial cytochrome b (CYTB), nuclear ribosomal 28S (28S) and intergenic spacer elongation factor-1 alpha ( EF-1 ALPHA) on 44 specimens collected from South Africa, four from Mozambique, and three each from single locations in India and Australia. Phylogenetic analyses were conducted using the Maximum Parsimony (MP) and Neighbour-Joining (NJ) methods. All of the datasets of the five DNA gene regions that were sequenced were also analyzed using the Basic Local Alignment Search Tool (BLAST) to find the closest matches for inclusion in the phylogenetic trees as outgroups and for purposes of information. In the phylogenetic trees for COI, COII, cytb and EF-1 ALPHA, a similar pattern was observed in the way that the sequences assembled into different groups; i.e., some sequences of A. simplexella from Australia were grouped separately from the others, but some Australian sequences grouped with those of the GLM from South Africa, India, and Mozambique. In the phylogenetic tree for 28S, all sequences from South Africa, Australia, India, and Mozambique grouped together and formed one group. For COI, genetic pairwise distance ranged from 0.97 to 3.60 %, for COII it ranged from 0.19% to 2.32%, for cytb it ranged from 0.25 to 9.77% and for EF-1 ALPHA it ranged 0.48 to 6.99%. Results of this study indicate that these populations are genetically related and presumably constitute a single species. Thus, further molecular and morphological studies need to be undertaken in order to resolve this apparent conundrum on the taxonomy of these populations.

Keywords: aproaerema modicella, aproaerema simplexella, mitochondrial DNA, nuclear DNA

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3152 An Interpretive Phenomenological Approach to Children’s Experience of Infidelity in Iran: Consequences and Hurts

Authors: Mehravar Javid, Stacey Dershewitz, Shohreh Hashemighoochani, Artin Yousefi

Abstract:

One of the most common reasons for destroying relationships is infidelity. This study was conducted to understand children's experience of infidelity in Iran. In this qualitative research, the interpretive phenomenological approach and theoretical (purposive) sampling method were used, and the data was saturated by conducting semi-structured interviews with eight offspring of families affected by infidelity. All interviews were audio recorded and analyzed on paper after implementation. Acceptability and homogeneity of data were confirmed by different methods. Eight main themes were identified from the participants' statements: description of the family atmosphere, lack of security and distrust, emotional reactions, behavioral reactions, worry about the future, financial problems, mother's physical-psychological state, and mother's coping strategies and offspring's proposed solutions. The children's experiences showed that in addition to causing irreparable harm to the wife, infidelity affected the whole family and caused the children to suffer crucial consequences such as emotional, behavioral, educational, and economic consequences and mistrust.

Keywords: consequences and hurt, infidelity, offspring, phenomenology, qualitative research

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3151 Dys-Regulation of Immune and Inflammatory Response in in vitro Fertilization Implantation Failure Patients under Ovarian Stimulation

Authors: Amruta D. S. Pathare, Indira Hinduja, Kusum Zaveri

Abstract:

Implantation failure (IF) even after the good-quality embryo transfer (ET) in the physiologically normal endometrium is the main obstacle in in vitro fertilization (IVF). Various microarray studies have been performed worldwide to elucidate the genes requisite for endometrial receptivity. These studies have included the population based on different phases of menstrual cycle during natural cycle and stimulated cycle in normal fertile women. Additionally, the literature is also available in recurrent implantation failure patients versus oocyte donors in natural cycle. However, for the first time, we aim to study the genomics of endometrial receptivity in IF patients under controlled ovarian stimulation (COS) during which ET is generally practised in IVF. Endometrial gene expression profiling in IF patients (n=10) and oocyte donors (n=8) were compared during window of implantation under COS by whole genome microarray (using Illumina platform). Enrichment analysis of microarray data was performed to determine dys-regulated biological functions and pathways using Database for Annotation, Visualization and Integrated Discovery, v6.8 (DAVID). The enrichment mapping was performed with the help of Cytoscape software. Microarray results were validated by real-time PCR. Localization of genes related to immune response (Progestagen-Associated Endometrial Protein (PAEP), Leukaemia Inhibitory Factor (LIF), Interleukin-6 Signal Transducer (IL6ST) was detected by immunohistochemistry. The study revealed 418 genes downregulated and 519 genes upregulated in IF patients compared to healthy fertile controls. The gene ontology, pathway analysis and enrichment mapping revealed significant downregulation in activation and regulation of immune and inflammation response in IF patients under COS. The lower expression of Progestagen Associated Endometrial Protein (PAEP), Leukemia Inhibitory Factor (LIF) and Interleukin 6 Signal Transducer (IL6ST) in cases compared to controls by real time and immunohistochemistry suggests the functional importance of these genes. The study was proved useful to uncover the probable reason of implantation failure being imbalance of immune and inflammatory regulation in our group of subjects. Based on the present study findings, a panel of significant dysregulated genes related to immune and inflammatory pathways needs to be further substantiated in larger cohort in natural as well as stimulated cycle. Upon which these genes could be screened in IF patients during window of implantation (WOI) before going for embryo transfer or any other immunological treatment. This would help to estimate the regulation of specific immune response during WOI in a patient. The appropriate treatment of either activation of immune response or suppression of immune response can be then attempted in IF patients to enhance the receptivity of endometrium.

Keywords: endometrial receptivity, immune and inflammatory response, gene expression microarray, window of implantation

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3150 Collagen Deposition in Lung Parenchyma Driven by Depletion of LYVE-1+ Macrophages Protects Emphysema and Loss of Airway Function

Authors: Yinebeb Mezgebu Dagnachew, Hwee Ying Lim, Liao Wupeng, Sheau Yng Lim, Lim Sheng Jie Natalie, Veronique Angeli

Abstract:

Collagen is essential for maintaining lung structure and function, and its remodeling has been associated with respiratory diseases, including chronic obstructive pulmonary disease (COPD). However, the cellular mechanisms driving collagen remodeling and the functional implications of this process in the pathophysiology of pulmonary diseases remain poorly understood. Using a mouse model of Lyve-1 expressing macrophage depletion, we found that the absence of this subpopulation of tissue-resident macrophage led to the preferential deposition of type I collagen fibers around the alveoli and bronchi in the steady state. Further analysis by polarized light microscopy revealed that the collagen fibers accumulating in the lungs depleted of Lyve-1+ macrophages were thicker and crosslinked. A decrease in MMP-9 gene expression and proteolytic activity, together with an increase in Col1a1, Timp-3 and Lox gene expression, accompanied the collagen alterations. Next, we investigated the effect of the collagen remodeling on the pathophysiology of COPD and airway function in mouse lacking Lyve-1+ macrophage exposed chronically to cigarette smoke (CS), a well-established animal model of COPD. We showed that the deposition of collagen protected mouse against the destruction of alveoli (emphysema) and bronchi thickening after CS exposure and prevented loss of airway function. Thus, we demonstrate that interstitial Lyve-1+ macrophages regulate the composition, amount, and architecture of the collagen network in the lungs and that such collagen remodeling functionally impacts the development of COPD. This study further supports the potential of targeting collagen as a promising approach to treating respiratory diseases.

Keywords: lung, extracellular matrix, chronic obstructive pulmonary disease, matrix metalloproteinases, collagen

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3149 An Endophyte of Amphipterygium adstringens as Producer of Cytotoxic Compounds

Authors: Karol Rodriguez-Peña, Martha L. Macias-Rubalcava, Leticia Rocha-Zavaleta, Sergio Sanchez

Abstract:

A bioassay-guided study for anti-cancer compounds from endophytes of the Mexican medicinal plant Amphipteryygium adstringens resulted in the isolation of a streptomycete capable of producing a group of compounds with high cytotoxic activity. Microorganisms from surface sterilized samples of various sections of the plant were isolated and all the actinomycetes found were evaluated for their potential to produce compounds with cytotoxic activity against cancer cell lines MCF7 (breast cancer) and HeLa (cervical cancer) as well as the non-tumoural cell line HaCaT (keratinocyte). The most active microorganism was picked for further evaluation. The identification of the microorganism was carried out by 16S rDNA gene sequencing, finding the closest proximity to Streptomyces scabrisporus, but with the additional characteristic that the strain isolated in this study was capable of producing colorful compounds never described for this species. Crude extracts of dichloromethane and ethyl acetate showed IC50 values of 0.29 and 0.96 μg/mL for MCF7, 0.51 and 1.98 μg/mL for HeLa and 0.96 and 2.7 μg/mL for HaCaT. Scaling the fermentation to 10 L in a bioreactor generated 1 g of total crude extract, which was fractionated by silica gel open column to yield 14 fractions. Nine of the fractions showed cytotoxic activity. Fraction 4 was chosen for subsequent purification because of its high activity against cancerous cell lines, lower activity against keratinocytes. HPLC-UV-MS/ESI was used for the evaluation of this fraction, finding at least 10 different compounds with high values of m/z (≈588). Purification of the compounds was carried out by preparative thin-layer chromatography. The prevalent compound was Steffimycin B, a molecule known for its antibiotic and cytotoxic activities and also for its low solubility in aqueous solutions. Along with steffimycin B, another five compounds belonging to the steffimycin family were isolated and at this moment their structures are being elucidated, some of which display better solubility in water: an attractive property for the pharmaceutical industry. As a conclusion to this study, the isolation of endophytes resulted in the discovery of a strain capable of producing compounds with high cytotoxic activity that need to be studied for their possible utilization.

Keywords: amphipterygium adstringens, cytotoxicity, streptomyces scabrisporus, steffimycin

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3148 Viability of Zoning Reform in Tackling Urban Inequality in Louisville

Authors: Mojeed A. Oladele

Abstract:

The original zoning system in Louisville promoted social segregation among groups and remained a tool for social exclusion that strengthened preexisting inequalities. The current residential zoning system in Louisville is predominantly single-family residential housing. Of the 75% of total land allocated for residential purposes, 55% comprises single-family housing, constituting one form of development and ruminant problems of social segregation within the city. The zoning reform initiative birthed the spatial improvement and development of additional middle housing as a more generic and inclusive housing form. The paper investigates the basis of zoning reform relative to the interconnectedness amongst the discursive objects of analysis and the extensiveness as a strategic tool of structural adjustment. Qualitative methodological assessment generated by collective planning professionals reflects the effectiveness of the new zoning design in strengthening the socio-spatial interactions within the city. The zoning reform is currently at the early stage of implementation and requires more professional/public inputs and constant iterative processes for a more promising urban planning outcome.

Keywords: zoning reform, viability, urban inequality, housing affordability, Louisville

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3147 Influence of Interpersonal Communication on Family Planning Practices among Rural Women in South East Nigeria

Authors: Chinwe Okpoko, Vivian Atasie

Abstract:

One of the leading causes of death amongst women of child-bearing age in southeast Nigeria is pregnancy. Women in the reproductive age group die at a higher rate than men of the same age bracket. Furthermore, most maternal deaths occur among poor women who live in rural communities, and who generally fall within the low socio-economic group in society. Failure of policy makers and the media to create the strategic awareness and communication that conform with the sensibilities of this group account, in part, for the persistence of this malaise. Family planning (FP) is an essential component of safe motherhood, which is designed to ensure that women receive high-quality care to achieve an optimum level of health of mother and infant. The aim is to control the number of children a woman can give birth to and prevent maternal and child mortality and morbidity. This is what sustainable development goal (SDG) health target of World Health Organization (WHO) also strives to achieve. FP programmes reduce exposure to the risks of child-bearing. Indeed, most maternal deaths in the developing world can be prevented by fully investing simultaneously in FP and maternal and new-born care. Given the intrinsic value of communication in health care delivery, it is vital to adopt the most efficacious means of awareness creation and communication amongst rural women in FP. In a country where over 50% of her population resides in rural areas with attendant low-level profile standard of living, the need to communicate health information like FP through indigenous channels becomes pertinent. Interpersonal communication amongst family, friends, religious groups and other associations, is an efficacious means of communicating social issues in rural Africa. Communication in informal settings identifies with the values and social context of the recipients. This study therefore sought to determine the place of interpersonal communication on the knowledge of rural women on FP and how it influences uptake of FP. Descriptive survey design was used in the study, with interviewer administered questionnaire constituting the instrument for data collection. The questionnaire was administered on 385 women from rural communities in southeast Nigeria. The results show that majority (58.5%) of the respondents agreed that interpersonal communication helps women understand how to plan their family size. Many rural women (82%) prefer the short term natural method to the more effective modern contraceptive methods (38.1%). Husbands’ approval of FP, as indicated in the Mean response of 2.56, is a major factor that accounts for the adoption of FP messages among rural women. Socio-demographic data also reveal that educational attainment and/or exposure influenced women’s acceptance or otherwise of FP messages. The study, therefore, recommends amongst others, the targeting of husbands in subsequent FP communication interventions, since they play major role on contraceptive usage.

Keywords: family planning, interpersonal communication, interpersonal interaction, traditional communication

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3146 Effect of Graded Level of Nano Selenium Supplementation on the Performance of Broiler Chicken

Authors: Raj Kishore Swain, Kamdev Sethy, Sumanta Kumar Mishra

Abstract:

Selenium is an essential trace element for the chicken with a variety of biological functions like growth, fertility, immune system, hormone metabolism, and antioxidant defense systems. Selenium deficiency in chicken causes exudative diathesis, pancreatic dystrophy and nutritional muscle dystrophy of the gizzard, heart and skeletal muscle. Additionally, insufficient immunity, lowering of production ability, decreased feathering of chickens and increased embryo mortality may occur due to selenium deficiency. Nano elemental selenium, which is bright red, highly stable, soluble and of nano meter size in the redox state of zero, has high bioavailability and low toxicity due to the greater surface area, high surface activity, high catalytic efficiency and strong adsorbing ability. To assess the effect of dietary nano-Se on performance and expression of gene in Vencobb broiler birds in comparison to its inorganic form (sodium selenite), four hundred fifty day-old Vencobb broiler chicks were randomly distributed into 9 dietary treatment groups with two replicates with 25 chicks per replicate. The dietary treatments were: T1 (Control group): Basal diet; T2: Basal diet with 0.3 ppm of inorganic Se; T3: Basal diet with 0.01875 ppm of nano-Se; T4: Basal diet with 0.0375 ppm of nano-Se; T5: Basal diet with 0.075 ppm of nano-Se, T6: Basal diet with 0.15 ppm of nano-Se, T7: Basal diet with 0.3 ppm of nano-Se, T8: Basal diet with 0.60 ppm of nano-Se, T9: Basal diet with 1.20 ppm of nano-Se. Nano selenium was synthesized by mixing sodium selenite with reduced glutathione and bovine serum albumin. The experiment was carried out in two phases: starter phase (0-3 wks), finisher phase (4-5 wk) in deep litter system. The body weight at the 5th week was best observed in T4. The best feed conversion ratio at the end of 5th week was observed in T4. Erythrocytic catalase, glutathione peroxidase and superoxide dismutase activity were significantly (P < 0.05) higher in all the nano selenium treated groups at 5th week. The antibody titers (log2) against Ranikhet diseases vaccine immunization of 5th-week broiler birds were significantly higher (P < 0.05) in the treatments T4 to T7. The selenium levels in liver, breast, kidney, brain, and gizzard were significantly (P < 0.05) increased with increasing dietary nano-Se indicating higher bioavailability of nano-Se compared to inorganic Se. The real time polymer chain reaction analysis showed an increase in the expression of antioxidative gene in T4 and T7 group. Therefore, it is concluded that supplementation of nano-selenium at 0.0375 ppm over and above the basal level can improve the body weight, antioxidant enzyme activity, Se bioavailability and expression of the antioxidative gene in broiler birds.

Keywords: chicken, growth, immunity, nano selenium

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3145 Sense of Involvement and Support in Persons with Cognitive Decline in Ordinary Dwelling

Authors: Annika Kjallman Alm, Ove Hellzen, Malin Rising-Holmstrom

Abstract:

Worldwide, the number of people who are living with dementia is increasing because of an aging population, which leads to increased financial and social costs, including reduced quality of life for people with dementia and their care partners. Most people who have dementia reside in the community. Aging in place could be described as having the health and social supports and services you need to live safely and independently in your home or your society for as long as you wish and are able. People with dementia are not different than people without dementia where they want to remain at home, if possible, with a sense of familiarity and engagement in typical everyday activities. So how do persons with dementia or cognitive decline see their possibilities to be socially involved and experience support? The aim of this study was to explore persons with cognitive decline's sense of involvement and support living in the ordinary dwelling. The study was approved by the Ethical Review Authority in Sweden prior to the interviews. Interviews were conducted with 20 persons living at home, either alone or in a relationship. The persons had perceived cognitive decline; some were under investigation or already had a diagnose of early dementia. Thematic analysis was used to identify, analyze, and report patterns within the data. Researchers extracted three main themes through participants’ interviews: a) Importance of social involvement with family and friends. b) Hindrances for social involvement. c) Struggling mentally with a new life situation. Results found that going to activity centers, staying involved, and meeting friends and family enhanced the sense of involvement and support. There were also hindrances to a sense of involvement and support as they struggled with the diagnose and the changes in daily life, such as physical problems, mental problems, or economic issues. The mental struggle of accepting the cognitive decline and the changes in daily life it brought was also an issue for some of the participants. A multidimensional support should be provided by the community to enable persons with cognitive decline to stay involved in family and community in the comfort of their own homes.

Keywords: aging in place, cognitive decline, dementia, sense of involvement

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3144 Reliability Analysis of Computer Centre at Yobe State University Using LRU Algorithm

Authors: V. V. Singh, Yusuf Ibrahim Gwanda, Rajesh Prasad

Abstract:

In this paper, we focus on the reliability and performance analysis of Computer Centre (CC) at Yobe State University, Damaturu, Nigeria. The CC consists of three servers: one database mail server, one redundant and one for sharing with the client computers in the CC (called as a local server). Observing the different possibilities of the functioning of the CC, the analysis has been done to evaluate the various popular measures of reliability such as availability, reliability, mean time to failure (MTTF), profit analysis due to the operation of the system. The system can ultimately fail due to the failure of router, redundant server before repairing the mail server and switch failure. The system can also partially fail when a local server fails. The failed devices have restored according to Least Recently Used (LRU) techniques. The system can also fail entirely due to a cooling failure of the server, electricity failure or some natural calamity like earthquake, fire tsunami, etc. All the failure rates are assumed to be constant and follow exponential time distribution, while the repair follows two types of distributions: i.e. general and Gumbel-Hougaard family copula distribution.

Keywords: reliability, availability Gumbel-Hougaard family copula, MTTF, internet data centre

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3143 Reducing Inequalities for the Uptake of Long-Term Reversible Contraceptive Methods through Special Family Planning Camps: A High Impact Service Delivery Model of Family Planning Practices

Authors: Ghulam Mustafa Halepota, Zaib Dahar

Abstract:

Background: Low acceptance of FP services, particularly in hard to reach areas where geographic, economic, or social barriers limit-service uptake. Moreover, limited resources appeared to be a reflection of dismal contraceptive use in Pakistan. People’s Primary Health Care Initiative (PPHI) is a Public Private Partnership Program of Government of Sindh which aims to improve maternal child health through accessible family planning services in far flung areas. In 2015 PPHI launched special family planning camps to have achieved a rapid improvement in CPR. On quarterly basis, these camps focus on Long Acting Reversible Contraceptives (LARC). These camps are arranged at 250 BHU Plus (24/7 MCHCs). The Organization manages 1140 primary health care facilities all over Sindh province and focuses on maternal, newborn and child health which includes antenatal care, labor/delivery, postnatal care, family planning, immunization, nutrition, BEmONC, CEmONC, diagnostic laboratories, ambulance services. Under the FPRH program, the organization launched special family planning camps in far flung areas to achieve a rapid improvement in CPR-committed to FP 2020 goal. Objective: To assess the performance of special FP camps for the improvement of long acting reversible contraceptive in hard to reach areas. Methodology: Outreach camps are organized on quarterly basis in 250 BHUs and maternal and child health centers (available-24/7). Using observational study design, the study reports 2 years data of special FP camps conducted in 23 various districts of Sindh during April 2015-April 2017. These special camps served a range of modern contraceptive methods including IUCDs, implants, condoms, pills, and injections. Moreover, 125 male medical officers are trained across Sindh in LARC and 554 female have been trained in implants and IUCD insertions. MSI Impact calculator was used to determine health and demographic impact of services. Results: This intervention has brought exceptional results, and the response has been overwhelming in time. Total 2048 special camps during 2015 till April 2017 have been carried out. 231796 MWRAs visited camps 91% opted modern FP, of which 45% opted Implants, 6% selected IUCDs from LARC (long term reversible contraceptive) from short term, 17% opted injectable 18% choose pills, and 12% used condoms. This intervention created a high contraceptive impact in rural Sindh an estimated 125048 FP users have been created, of this 111846 LARC users and 13498 are SARC users, through this intervention an estimated 55774 unintended pregnancies, 36299 live births, 9394, 80 maternal deaths, 926 and 6077 unsafe abortion have been averted. Moreover, the intervention created an economic impact and saved 2,409,563 direct health expenditure on each woman with reproductive age. Conclusion: Special FP Camps along with routine services is an effective and acceptable model for increase in provision of long-acting and permanent methods in hard to reach areas. This innovative approach by PHHI-Sindh has also been adopted in other provinces of Pakistan.

Keywords: inequalities, special camps, family planning services, hard to reach areas

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3142 Managing Children with Autism Spectrum Disorder in Corona Age

Authors: Raju Singh, Shikha Singh

Abstract:

This article is note for managing Autistic Child during the Corona time line. It becomes very critical for the primary caregiver as corona pandemic poses new challenges and completely variety of threats to line of treatment, growth, socialization, mental health for children with autism spectrum disorder (ASD), and, so for the family of the children. It is a highly distressful situation, where the line of treatment has shrunken, physical contact has reduced and therapies footprints reduced in several parts of the world. As children with ASD already face socialization challenges, isolation rules imposed by individuals (or social groups), government agencies have only made the situation worse for the children with ASD and their family. This note will try to touch the basics on understanding the ASD and related development disorders, challenges, impact, and suggest approaches to deal with such situation. This document also covers data analysis, deep dive into the increasing impact of ASD on children. This document can also act as a baseline for many researchers, psychiatrists, psychologists, therapists to view the problem statement and measure its impact.

Keywords: autism spectrum disorder, mental health, applied behavior therapy, occupational therapy, social anxiety

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3141 A Contrastive Analysis on Hausa and Yoruba Adjectival Phrases

Authors: Abubakar Maikudi

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Contrastive analysis is the method of analyzing the structure of any two languages with a view to determining the possible differential aspects of their systems irrespective of their genetic affinity or level of development. Contrastive analysis of two languages becomes useful when it is adequately describing the sound structure and grammatical structure of two languages, with comparative statements giving emphasis to the compatible items in the two systems. This research work uses comparative analysis theory to analyze adjective and adjectival phrases in Hausa and Yorùbá languages. The Hausa language belongs to the Chadic family of the Afro-Asiatic phylum, while the Yorùbá language belongs to the Benue-Congo family of the Niger-Congo phylum. The findings of the research clearly demonstrated that there are significant similarities in the adjectival phrase constructions of the two languages, i.e., nominal (Head) and post-nominal (Post-Head) use of the adjective, predicative function of an adjective, use of the reduplicative adjective, use of the comparative and superlative adjective, etc. However, there are dissimilarities in the adjectival phrase of the two languages in gender/number agreement and pre-nominal (Post-Head) use of adjectives.

Keywords: genetic affinity, contrastive analysis, phylum, pre-head, post-head

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3140 RNA-seq Analysis of Liver from NASH-HCC Model Mouse Treated with Streptozotocin-High Fat Diet

Authors: Bui Phuong Linh, Yuki Sakakibara, Ryuto Tanaka, Elizabeth H. Pigney, Taishi Hashiguchi

Abstract:

Non-alcoholic steatohepatitis (NASH) is a chronic liver disease, often associated with type II diabetes, which sometimes progresses to more serious conditions such as liver fibrosis and hepatocellular carcinoma (HCC). NASH has become an important health problem worldwide, buttherapeutic agents for NASH have not yet been approved, and animal models with high clinical correlation are required. TheSTAM™ mouse shows the same pathological progression as human NASH patients and has been widely used for both drug efficacy and basic research, such as lipid profiling and gut microbiota research. In this study, we analyzed the RNA-seq data of STAM™mice at each pathological stage (steatosis, steatohepatitis, liver fibrosis, and HCC) and examined the clinical correlation at the genetic level. NASH was induced in male mice by a single subcutaneous injection of 200 µg streptozotocin solution 2 days after birth and feeding with high fat dietafter 4 weeks of age. The mice were sacrificed and livers collected at 6, 8, 10, 12, 16, and 20 weeks of age. For liver samples, the left lateral lobe was snap frozen in liquid nitrogen and stored at -80˚C for RNA-seq analysis. Total RNA of the cells was isolated using RNeasy mini kit. The gene expression of the canonical pathways in NASH progression from steatosis to hepatocellular carcinoma were analyzed, such as immune system process, oxidation-reduction process, lipid metabolic process. Moreover, since it has been reported that genetic traits are involved in the development of NASH-HCC, we next analyzed the genetic mutations in the STAM™mice. The number of individuals showing mutations in Mtorinvolved in Insulin signaling increases as the disease progresses, especially in the liver cancer phase. These results indicated a clinical correlation of gene profiles in the STAM™mouse.

Keywords: steatosis, non-alcoholic steatohepatitis, fibrosis, hepatocellular carcinoma, RNA-seq

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3139 Emotional Disclosure as Mediator Between Marital Satisfaction and Mental Health Problems in Women with Infertility

Authors: Sadia Saleem

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Infertility is a global health concern that may have a long-lasting effect on the psychosocial functioning of an individual. Rich research evidence has shown that women with infertility are at greater risk of adverse psychological experiences than men. The culture plays a risk factor when it comes to infertility. Family is considered as a central focus of a collectivistic culture like Pakistan and having children is the key factor that determines the quality of a marital relationship, individual well-being and overall standing in the society. In this collectivistic cultural context, women usually get the blame and experience more psychological distress and social isolation. A total sample of 121 (M 28.17, SD 4.73) women with primary infertility selected through purposive sampling were tested using Emotional Disclosure Questionnaire, Couple Satisfaction Index and Depression Anxiety Stress Scale. The results indicate that negative emotional disclosure positively mediates the relationship between marital satisfaction and mental health problems (p < .001) in women with primary infertility. The results are discussed in terms of psychosocial counseling and family psychoeducation in Pakistani collectivistic cultural context.

Keywords: infertility, couple satisfaction, emotional disclosure, mental health

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3138 Child Labour and the Challenges of National Security in Nigeria: A Study of Aba Urban, Abia State, Nigeria

Authors: Anyaogu Paul

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The study examines the problem of child labour in respect to National security in Nigeria. The current situation reflects a chronic urban poverty, which can compel parents and guardians to send their children and wards of school age to engage in income yielding activities to augment family income. The study sought to explain the basis of child labour, its causes and its effects on its victim and society at large. A social survey research design was employed to select the respondents. A sample size of 250 respondents was selected from targeted population of children below the age of 18 years. A questionnaire instrument was employed to collect data. An accidental sampling technique was employed to select the respondents. The findings revealed that child labour is on the increase and a serious threat to national security and social cohesion. The study recommends that the Nigerian government should enforce the laws on child labour and provide opportunities for job creation for urban dwellers. More so, government should also provide free and compulsory education at primary and secondary school levels of education.

Keywords: child labour, family income, national security, survey research

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3137 The Extent of Proliferation, Apoptosis and Angiogenesis at the Site of Injury Determine the Course of Healing Either as Scar Free or as Scarred One in the Appendages of Lizard

Authors: Isha Ranadive, Sonam Patel, Suresh Balakrishnan

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It has been observed that in lizards wound can be healed by either a scar free mechanism or by scarring. The animal model used to study both these healing processes was Northern House Gecko. In lizard, the tail when amputated heals by scar free mechanism which allows it to regenerate, the same is not seen when the limb is amputated. Proliferation, apoptosis, and angiogenesis are the main events which succeed an injury. We observed that proliferation of the cells beneath the wound epidermis was much higher in case of wound healing in tail. This could be because after the wound gets covered by the epithelium, it enters in to a cross-talk with the underlying mesenchyme to recruit a pool of blastemal cells which proliferate and later differentiate to form the lost part through epimorphic regeneration. This was substantiated by mRNA expression levels of various FGFs which facilitate the cross-talk and also by PCNA which is a marker for proliferation. Western blot result reaffirms the same notion. However, in case of the limb, the rate of apoptosis was more than proliferation as there are a lot of debris that needs to be removed. We came to this conclusion as we observed that p53 the apoptotic gene was highly upregulated in case of the scarred tissue. Further, we confirmed this result by checking the anti-apoptotic gene bcl2 and found it to be significantly down-regulated. As we noticed heightened proliferation in the case of scar-free wound healing in tail, angiogenesis was targeted for the study. This is because, when the cells are proliferating they require constant supply of blood and hence neo-vascularization is inevitable. It was observed that the marker of angiogenesis, VEGF, was expressed more during wound healing as compared to the resting stage of tail. Moreover, a high up-regulation was seen in KDR, a receptor of VEGF. Thus, this study reveals how proliferation, apoptosis, and angiogenesis play a key role in the scar-free as well as scarred wound healing.

Keywords: epimorphic regeneration, injury, northern house gecko, wound healing

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3136 Effects of Delphinidin on Lipid Metabolism in HepG2 Cells and Diet-Induced Obese Mice

Authors: Marcela Parra-Vargas, Ana Sandoval-Rodriguez, Roberto Rodriguez-Echevarria, Jose Dominguez-Rosales, Juan Armendariz-Borunda

Abstract:

Non-alcoholic fatty liver disease (NAFLD) is characterized by an excess of hepatic lipids, and it is to author’s best knowledge, the most prevalent chronic liver disorder. Anthocyanin-rich food consumption is linked to health benefits in metabolic disorders associated with obesity and NAFLD, although the precise functional role of anthocyanidin delphinidin (Dp) has yet to be established. The aim of this study was to investigate the effect of the Dp in NAFLD metabolic alterations by evaluating prevention or amelioration of hepatic lipid accumulation, as well as molecular mechanisms in two experimental obesity-related models of NALFD. In vitro: HepG2 cells were incubated with sodium palmitate (PA, 1 mM) to induce lipotoxic damage, and concomitantly treated with Dp (180 uM) for 24 h. Subsequently, total lipid accumulation was measured by colorimetric staining with Oil Red O, and total intrahepatic triglycerides were determined by an enzymatic assay. To assess molecular mechanisms, cells were pre-treated with PA for 24 h and then exposed to Dp for 1 h. In vivo: four-week-old male C57BL/6Nhsd mice were allocated in two main groups. Mice were fed with standard diet (control) or high-fat and high-carbohydrate diet (45% fat, HFD) for 16 wk to induce NAFLD. Then HFD was divided into subgroups: one treated orally with Dp (15 mg/kg bw, HFD-Dp) every day for 4 wk, while HFD group treated with vehicle (DMSO). Weight and fasting glucose were recorded weekly, while dietary ingestion was measured daily. Insulin tolerance test was performed at the end of treatment. Liver histology was evaluated with H&E and Masson’s trichrome stain. RT-PCR was used to evaluate gene expression and Western Blot to determine levels of protein in both experimental models. Parametric data were analyzed with one-way ANOVA and Tukey’s post-hoc test. Kruskal-Wallis and Mann-Whitney U test for non-parametric data, and P < 0.5 were considered significant. Dp prevented hepatic lipid accumulation by PA in HepG2 hepatocytes. Furthermore, Dp down-regulated gene expression of SREBP1c, FAS, and CPT1a without modifying AMPK phosphorylation levels. In vivo, Dp oral administration did not ameliorate lipid metabolic alterations raised by HFD. Adiposity, dietary ingestion, fasting glucose, and insulin sensitivity after Dp treatment remained similar to HFD group. Histological analysis showed hepatic damage in HFD groups and no differences between HFD and HFD-Dp groups were found. Hepatic gene expression of ACC and FAS were not altered by HFD. SREBP1c was similar in both HFD and HFD-Dp groups. No significant changes were observed in SREBP1c, ACC, and FAS adipose tissue gene expression by HFD or Dp treatment. Additionally, immunoblotting analysis revealed no changes in pathway SIRT1-LKB-AMPK and PPAR alpha by both HFD groups compared to control. In conclusion, the antioxidant Dp may provoke beneficial effects in the prevention of hepatic lipid accumulation. Nevertheless, the oral dose administrated in mice that simulated the total intake of anthocyanins consumed daily by humans has no effect as a treatment on hepatic lipid metabolic alterations and histological abnormalities associated with exposure to chronic HFD. A healthy lifestyle with regular intake of antioxidants such as anthocyanins may prevent metabolic alterations in NAFLD.

Keywords: anthocyanins, antioxidants, delphinidin, non-alcoholic fatty liver disease, obesity

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3135 Functional Switching of Serratia marcescens Transcriptional Regulator from Activator to Inhibitor of Quorum Sensing by Exogenous Addition

Authors: Norihiro Kato, Yuriko Takayama

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Some gram-negative bacteria enable the simultaneous activation of gene expression involved in N-acylhomoserine lactone (AHL) dependent cell-to-cell communication system. Such regulatory system for the bacterial group behavior is termed as quorum sensing (QS) because a diffusible AHL signal can accumulate around the cell during the increase of the cell density and trigger activation of the sequential QS process. By blocking the QS, the expression of diverse genes related to infection, antibiotic production, and biofilm formation is inhibited. Conditioning of QS by regulation of the DNA-receptor-AHL interaction is a potential target for enhancing host defenses against pathogenicity. We focused on engineered application of transcriptional regulator SpnR produced in opportunistic human pathogen Serratia marcescens. The SpnR can interact with AHL signals at an N-terminal domain and also with a promoter region of a QS target gene at a C-terminal domain. As the initial process of the QS activation, the SpnR forms a complex with the AHL to enhance the expression of pig cluster; the SpnR normally acts as an activator for the expression of the QS-dependent gene. In this research, we attempt to artificially control QS by changing the role of SpnR. The QS-dependent prodigiosin production is expected to inhibit by externally added SpnR in the culture broth of AS-1 strain because the AHL concentration was kept below the threshold by AHL-SpnR complex formation. Maltose-binding protein (MBP)-tagged SpnR (MBP-SpnR) was overexpressed in Escherichia coli and purified using an affinity chromatography equipped with an amylose resin column. The specific interaction between AHL and MBP-SpnR was demonstrated by quartz crystal microbalance (QCM) sensor. AHL with amino end-group was coupled with COOH-terminated self-assembled monolayer prepared on a gold electrode of 27-MHz quartz crystal sensor using water-soluble carbodiimide. After the injection of MBP-SpnR into a cup-type sensor cell filled with the buffer solution, time course of resonant frequency change (ΔFs) was determined. A decrease of ΔFs clearly showed the uptake of MBP-SpnR onto the AHL-immobilized electrode. Furthermore, no binding affinity was observed after the heat-inactivation of MBP-SpnR at 80ºC. These results suggest that MBP-SpnR possesses a specific affinity for AHL. MBP-SpnR was added to the culture medium as an AHL trap to study inhibitory effects on intracellularly accumulated prodigiosin. With approximately 2 µM MBP-SpnR, the amount of prodigiosin induced was half that of the control without any additives. In conclusion, the function of SpnR could be switched by adding it to the cell culture. Exogenously added MBP-SpnR possesses high affinity for AHL derived from cells and acts as an inhibitor of AHL-mediated QS.

Keywords: intracellular signaling, microbial biotechnology, quorum sensing, transcriptional regulator

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3134 Migrants as Change Agents: A Study of Social Remittances between Finland and Russia

Authors: Ilona Bontenbal

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In this research, the potential for societal change is researched through the idea of migrants as change agents. The viewpoint is on the potential that migrants have for affecting societal change in their country of origin through transmitting transnational peer-to-peer information. The focus is on the information that Russian migrants living in Finland transmit about their experiences and attitudes regarding the Nordic welfare state, its democratic foundation and the social rights embedded in it, to their family and friends in their country of origin. The welfare provision and level of democracy are very different in the two neighbouring countries of Finland and Russia. Finland is a Nordic welfare state with strong democratic institutions and a comprehensive actualizing of civil and social rights. In Russia, the state of democracy has on the other hand been declining, and the social and civil rights of its citizens are constantly undermined. Due to improvements in communications and travel technology, migrants can easily and relatively cheaply stay in contact with their family and friends in their country of origin. This is why it is possible for migrants to act as change agents. By telling about their experiences and attitudes about living in a democratic welfare state, migrants can affect what people in the country or origin know and think about welfare, democracy, and social rights. This phenomenon is approached through the concept of social remittances. Social remittances broadly stand for the ideas, know-how, world views, attitudes, norms of behavior, and social capital that flows through transnational networks from receiving- to sending- country communities and the other way around. The viewpoint is that historically and culturally formed democratic welfare models cannot be copied entirely nor that each country should achieve identical development paths, but rather that migrants themselves choose which aspects they see as important to remit to their acquaintances in their country of origin. This way the potential for social change and the agency of the migrants is accentuated. The empirical research material of this study is based on 30 qualitative interviews with Russian migrants living in Finland. Russians are the largest migrant group in Finland and Finland is a popular migration destination especially for individuals living in North-West Russia including the St. Petersburg region. The interviews are carried out in 2018-2019. The preliminary results indicate that Russian migrants discuss social rights and welfare a lot with their family members and acquaintances living in Russia. In general, the migrants feel that they have had an effect on the way that their friends and family think about Finland, the West, social rights and welfare provision. Democracy, on the other hand, is seen as a more difficult and less discussed topic. The transformative potential that the transmitted information and attitudes could have outside of the immediate circle of acquaintances on larger societal change is seen as ambiguous although not negligible.

Keywords: migrants as change agents, Russian migrants, social remittances, welfare and democracy

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3133 The Relationship between Violence against Women and Levels of Self-Esteem in Urban Informal Settlements of Mumbai, India: A Cross-Sectional Study

Authors: A. Bentley, A. Prost, N. Daruwalla, D. Osrin

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Background: This study aims to investigate the relationship between experiences of violence against women in the family, and levels of self-esteem in women residing in informal settlement (slum) areas of Mumbai, India. The authors hypothesise that violence against women in Indian households extends beyond that of intimate partner violence (IPV), to include other members of the family and that experiences of violence are associated with lower levels of self-esteem. Methods: Experiences of violence were assessed through a cross-sectional survey of 598 women, including questions about specific acts of emotional, economic, physical and sexual violence across different time points, and the main perpetrator of each. Self-esteem was assessed using the Rosenberg self-esteem questionnaire. A global score for self-esteem was calculated and the relationship between violence in the past year and Rosenberg self-esteem score was assessed using multivariable linear regression models, adjusted for years of education completed, and clustering using robust standard errors. Results: 482 (81%) women consented to interview. On average, they were 28.5 years old, had completed 6 years of education and had been married 9.5 years. 88% were Muslim and 46% lived in joint families. 44% of women had experienced at least one act of violence in their lifetime (33% emotional, 22% economic, 24% physical, 12% sexual). Of the women who experienced violence after marriage, 70% cited a perpetrator other than the husband for at least one of the acts. 5% had low self-esteem (Rosenberg score < 15). For women who experienced emotional violence in the past year, the Rosenberg score was 2.6 points lower (p < 0.001). It was 1.2 points lower (p = 0.03) for women who experienced economic violence. For physical or sexual violence in the past year, no statistically significant relationship with Rosenberg score was seen. However, for a one-unit increase in the number of different acts of each type of violence experienced in the past year, a decrease in Rosenberg score was seen (-0.62 for emotional, -0.76 for economic, -0.53 for physical and -0.47 for sexual; p < 0.05 for all). Discussion: The high prevalence of violence experiences across the lifetime was likely due to the detailed assessment of violence and the inclusion of perpetrators within the family other than the husband. Experiences of emotional or economic violence in the past year were associated with lower Rosenberg scores and therefore lower self-esteem, but no relationship was seen between experiences of physical or sexual violence and Rosenberg score overall. For all types of violence in the past year, a greater number of different acts were associated with a decrease in Rosenberg score. Emotional violence showed the strongest relationship with self-esteem, but for all types of violence the more complex the pattern of perpetration with different methods used, the lower the levels of self-esteem. Due to the cross-sectional nature of the study causal directionality cannot be attributed. Further work to investigate the relationship between severity of violence and self-esteem and whether self-esteem mediates relationships between violence and poorer mental health would be beneficial.

Keywords: family violence, India, informal settlements, Rosenberg self-esteem scale, self-esteem, violence against women

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3132 Biochemical Characterization of CTX-M-15 from Enterobacter cloacae and Designing a Novel Non-β-Lactam-β-Lactamase Inhibitor

Authors: Mohammad Faheem, M. Tabish Rehman, Mohd Danishuddin, Asad U. Khan

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The worldwide dissemination of CTX-M type β-lactamases is a threat to human health. Previously, we have reported the spread of blaCTX-M-15 gene in different clinical strains of Enterobacteriaceae from the hospital settings of Aligarh in north India. In view of the varying resistance pattern against cephalosporins and other β-lactam antibiotics, we intended to understand the correlation between MICs and catalytic activity of CTX-M-15. In this study, steady-state kinetic parameters and MICs were determined on E. coli DH5α transformed with blaCTX-M-15 gene that was cloned from Enterobacter cloacae (EC-15) strain of clinical background. The effect of conventional β-lactamase inhibitors (clavulanic acid, sulbactam and tazobactam) on CTX-M-15 was also studied. We have found that tazobactam is the best among these inhibitors against CTX-M-15. The inhibition characteristic of tazobactam is defined by its very low IC50 value (6 nM), high affinity (Ki = 0.017 µM) and better acylation efficiency (k+2/K9 = 0.44 µM-1s-1). It forms an acyl-enzyme covalent complex, which is quite stable (k+3 = 0.0057 s-1). Since increasing resistance has been reported against conventional b-lactam antibiotic-inhibitor combinations, we aspire to design a non-b-lactam core containing b-lactamase inhibitor. For this, we screened ZINC database and performed molecular docking to identify a potential non-β-lactam based inhibitor (ZINC03787097). The MICs of cephalosporin antibiotics in combination with this inhibitor gave promising results. Steady-state kinetics and molecular docking studies showed that ZINC03787097 is a reversible inhibitor which binds non-covalently to the active site of the enzyme through hydrogen bonds and hydrophobic interactions. Though, it’s IC50 (180 nM) is much higher than tazobactam, it has good affinity for CTX-M-15 (Ki = 0.388 µM). This study concludes that ZINC03787097 compound can be used as seed molecule to design more efficient non-b-lactam containing b-lactamase inhibitor that could evade pre-existing bacterial resistance mechanisms.

Keywords: ESBL, non-b-lactam-b-lactamase inhibitor, bioinformatics, biomedicine

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3131 Role of Television in Constructing Gender for Young Women

Authors: Bhavna Negi

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Several studies highlight the significance of media in constructing realities around us. According to Forbes magazine the demand of televisions has increased several times in the developing nations. A recent survey reveals that 112 million Indian households have a television, with 61 percent accessing cable. The space and visibility of television has enormously grown over the last decade in Indian homes. This small box has indeed taken a large place in their daily routines. The multi channel viewing and TRPs puzzle the Indian audience. This medium creates and constructs social images and roles which form internal representation about societal functioing. According to National Council of Applied Economic Research about twenty seven percent Indian literate youth watches TV for recreation. The present study finds about the role of television and its impact on young college going women with reference to family based serials shown on television. It is interesting to see how young women perceive the popular family soaps and define norms, roles and spaces for a woman and a man. The paper further examines the subtle messages given to young women through television serials. It draws insights into the relationship between the contemporary Indian women and the images conceptualized and communicated on television.

Keywords: media, women, gender, social roles

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3130 Projects and Limits of Memory Engineering: A Case of Lithuanian Partisan War

Authors: Mingaile Jurkute, Vilnius University

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The memory of the Lithuanian partisan war (1944-1953) underwent extremely dramatic transformations. During this war, the image of the resistance and a partisan was one of the key elements of Lithuanian identity. Its importance is evidenced by the extremely large legacy of songs about partisans, no other topic has collected so much folklore in Lithuania. In the Soviet years, this resistance was practically forced to be forgotten. Terror and Soviet laws have forced people to stop talking about the events, even in the family circle. In addition, the Soviets created their own propaganda story, reinterpreting the Lithuanian partisan war, presenting partisans as bandits who brutally tortured and murdered locals. But even in the Soviet years, the memory could neither be completely suppressed, nor completely transformed into wishful shape. The analysis of fiction and cinema shows that the traumatic memory of real events rushed to the surface, thus transforming the very propagandistic narrative. After the restoration of the Republic of Lithuania in 1990, the Lithuanian partisan war was gradually returned to the central place of Lithuanian history. After 2014 the nationalist heroic narrative about Lithuanian partisans became the central narrative of modern Lithuanian history. Nevertheless, interviews I conducted in Lithuanian villages reveal that the memory of local communities and families preserves quite different experiences that do not fit into neither the Soviet narrative nor the heroic one. Such experiences include, for example, partisan violence against local families. This paper is about the efforts of two political ideologies (the Soviet and the Lithuanian patriotic) to use the history of the Lithuanian partisans for their own needs, and the attempts of small communities (mostly families) to resist these efforts. The research reveals that family memory, even when opposed to aggressive state memory policies, can preserve counter-narratives by exploiting unexpected objects beyond the control of the state, such as nature and wildlife. Basically, the paper analyses the limits of the instrumentalization of memory, even by extremely aggressive political regimes.

Keywords: collective memory, post-memory, violence, military conflict, family memory

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3129 Investigating the Impact of Factors Associated with Student Academic Achievement and Expectations through the Ecosystemic Perspective in the Greek Context: The Role of the Individual, Family, School and of the Community

Authors: Olga Giovani

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In this research, Bronfenbrenner's theory will be used to investigate the individual, microsystemic, and exosystemic factors that may affect adolescents' academic achievement as well as their expectations in Greece. First, the topic of academic achievement in an adolescent developmental context will be set as the target of the proposed study while focusing on the aspects of community influences on adolescents. More specifically, the effect of available resources and the perceived sense of safety and support will be further investigated. Then the issue of family factors will be analyzed, as they are subjectively perceived by the adolescents, focusing on the perceived parental style, parental monitor, and involvement as a mesosystemic factor. In turn, the school will also be discussed with emphasis on the perceived school climate and support as well as the academic aspects of student achievement. Finally, the adolescent's individual perspective will be taken into consideration in developmental terms, examining their perceptions regarding their community/neighborhood, their family, their school, as well as their sense of self-concept and self-esteem as these are expressed through their academic performance and prosocial behavior. The aim of the proposed research is to study these associations through the prism of the systemic perspective, the relationship between aspects of educational achievement and socioeconomic background, with an emphasis on the role of the community, which has not been adequately researched in the Greek context. Community will be defined by the available community resources (recreational activities, public library, local orchestras, free entrance museums, etc.), adolescents' own perception of social support, safety, and support inside that community. These perceptions need to be investigated since they may serve as possible predictors of a child's current cognitive, developmental, and psycho-social outcomes, such as their perceived self-concept and self-esteem, as well as on their future expectations related to the entrance to university and job expectations.

Keywords: bioecological model, developmental psychology, ecosystemic approach, student achievement, microsystemic factors, mesosystemic factors, individual perceptions

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3128 Fabrication of Electrospun Green Fluorescent Protein Nano-Fibers for Biomedical Applications

Authors: Yakup Ulusu, Faruk Ozel, Numan Eczacioglu, Abdurrahman Ozen, Sabriye Acikgoz

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GFP discovered in the mid-1970s, has been used as a marker after replicated genetic study by scientists. In biotechnology, cell, molecular biology, the GFP gene is frequently used as a reporter of expression. In modified forms, it has been used to make biosensors. Many animals have been created that express GFP as an evidence that a gene can be expressed throughout a given organism. Proteins labeled with GFP identified locations are determined. And so, cell connections can be monitored, gene expression can be reported, protein-protein interactions can be observed and signals that create events can be detected. Additionally, monitoring GFP is noninvasive; it can be detected by under UV-light because of simply generating fluorescence. Moreover, GFP is a relatively small and inert molecule, that does not seem to treat any biological processes of interest. The synthesis of GFP has some steps like, to construct the plasmid system, transformation in E. coli, production and purification of protein. GFP carrying plasmid vector pBAD–GFPuv was digested using two different restriction endonuclease enzymes (NheI and Eco RI) and DNA fragment of GFP was gel purified before cloning. The GFP-encoding DNA fragment was ligated into pET28a plasmid using NheI and Eco RI restriction sites. The final plasmid was named pETGFP and DNA sequencing of this plasmid indicated that the hexa histidine-tagged GFP was correctly inserted. Histidine-tagged GFP was expressed in an Escherichia coli BL21 DE3 (pLysE) strain. The strain was transformed with pETGFP plasmid and grown on LuiraBertoni (LB) plates with kanamycin and chloramphenicol selection. E. coli cells were grown up to an optical density (OD 600) of 0.8 and induced by the addition of a final concentration of 1mM isopropyl-thiogalactopyranoside (IPTG) and then grown for additional 4 h. The amino-terminal hexa-histidine-tag facilitated purification of the GFP by using a His Bind affinity chromatography resin (Novagen). Purity of GFP protein was analyzed by a 12 % sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The concentration of protein was determined by UV absorption at 280 nm (Varian Cary 50 Scan UV/VIS spectrophotometer). Synthesis of GFP-Polymer composite nanofibers was produced by using GFP solution (10mg/mL) and polymer precursor Polyvinylpyrrolidone, (PVP, Mw=1300000) as starting materials and template, respectively. For the fabrication of nanofibers with the different fiber diameter; a sol–gel solution comprising of 0.40, 0.60 and 0.80 g PVP (depending upon the desired fiber diameter) and 100 mg GFP in 10 mL water: ethanol (3:2) mixtures were prepared and then the solution was covered on collecting plate via electro spinning at 10 kV with a feed-rate of 0.25 mL h-1 using Spellman electro spinning system. Results show that GFP-based nano-fiber can be used plenty of biomedical applications such as bio-imaging, bio-mechanic, bio-material and tissue engineering.

Keywords: biomaterial, GFP, nano-fibers, protein expression

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3127 Silica Nanoparticles Induced Oxidative Stress and Inflammation in MRC-5 Human Lung Fibroblasts

Authors: Anca Dinischiotu, Sorina Nicoleta Voicu

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Silica nanoparticles (SiO2-NPs) are widely used in consumer products such as paints, plastics, insulation materials, tires, concrete production, as well as in gene delivery systems and imaging procedures. Environmental human exposure to them occurs during utilization of these products, in a time-dependent manner, the uptake being by topic and inhalation route especially. SiO2-NPs enter cells and induce membrane damage, oxidative stress and inflammatory reactions in a concentration-dependent manner. In this study, MRC-5 cells (human fetal lung fibroblasts) were exposed to amorphous SiO2-NPs at a dose of 62.5 μg/ml for 24, 48 and 72 hours. The size distribution of NPs was a lognormal function, in the range 3-14 nm. A time-dependent decrease of total reduced glutathione concentration by 36%, 50%, and 78% and an increase of NO level by 62%, 32%, respectively 24% compared to control were noticed. An up-regulation of NF-kB expression by 20%, 50% respectively 10% and of Nrf-2 by 139%, 58%, and 16% compared to control after 24, 48 and 72 hours was noticed also. The expression of IL-1β, IL-6, IL-8, and COX-2 was up-regulated in a time-dependent manner. Also, the expression of MMP-2 and MMP-9 were down-regulated after 48 and 72 hours, whereas their activities raised in a time-dependent manner. Exposure of cells to NPs up-regulated the expression of inducible NO synthase, as previously was shown, and probably this is the reason for the increased level of NO, that can react with the thiol groups of reduced glutathione molecules, diminishing its concentration Nrf2 is a transcription factor translocated in nucleus, under oxidative stress, where downstream gene expression activates in order to modulate the adaptive intracellular response against oxidative stress. The cross-talk between Nrf2 and NF-kB activities regulates the inflammatory processes. The activation of NF-kB could activate up-regulation of IL-1β, IL-6, and IL-8. The increase of COX-2 expression could be correlated with IL-1β one. Also, probably in response to the pro-inflammatory cytokines, MMP-2 and MMP-9 were induced and activated. In conclusion, the exposure of MRC-5 cells to SiO2-NPs generated inflammation in a time-dependent manner.

Keywords: inflammation, MRC-5 cells, oxidative stress, silica nanoparticles

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3126 First Report of Rahnella Victoriana Associated with Walnut Decline

Authors: Mohammadreza Hajialigol, Nargues Falahi Charkhabi, Fatemeh Shahryari, Saadat Sarikhani

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BACKGROUND AND OBJECTIVES Iran is the third producer of Persian walnut worldwide. However, its walnut trees have been under threat from decline during last decade. Walnut canker caused by B. nigrifluens and B. rubrifaciens was recorded in multiple regions of Iran. Furthermore, Brenneria rosae subsp. rosae and Gibbsiella quercinecans were recently recognized as responsible for walnut decline in northwestern Iran. This study aimed to identify the causal agent of walnut decline in Kermanshah and Isfahan. MATERIAL AND METHODS Symptomatic samples were collected from affected walnut trees of Kermanshah and Isfahan provinces. The pathogenicity of strains was proved on immature walnut fruits cv. ‘Hartley’ and young green twigs of two-year-old walnut seedling cv. ‘Chandler’. Pathogenic strains were subjected to conventional phenotypic tests. 16S rRNA, gyrB, and infB genes were partially amplified and sequenced. RESULTS Irregular longitudinal cankers and dark lesions were observed in the outer and inner bark, respectively. Twenty-four strains were isolated on EMB-agar media. Fourteen strains were able to cause necrosis and a dark-colored region in the mesocarp and on young green twigs around the inoculation site 14 and 30 days post-inoculation, respectively. Strains were able to hydrolyze Tween 20, Tween 80, gelatin and esculin, however, did not produce indole or urease. Pairwise comparison, the 16S rRNA gene nucleotide sequences of strain I2 were 100% identical with those of Rahnella victoriana FRB 225T. Moreover, a phylogenetic tree reconstructed based on the concatenated sequences of two housekeeping gene fragments, gyrB (601 bp) and infB (615 bp), revealed that the strains I2, I5, and KE6 were clustered with R. victoriana FRB 225T. CONCLUSION To the best of our knowledge, this is the first report of R. victoriana in association with walnut decline. This result is necessary to find resistant genotypes.

Keywords: emerging pathogens, Iran, juglans regia, MLSA

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3125 Biodegradation of 2,4-Dichlorophenol by Pseudomonas chlororaphis Strain Isolated from Activated Sludge Sample from a Wastewater Treatment Plant in Durban, South Africa

Authors: Boitumelo Setlhare, Mduduzi P. Mokoena, Ademola O. Olaniran

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Agricultural and industrial activities have led to increasing production of xenobiotics such as 2,4-dichlorophenol (2,4-DCP), a derivative of 2,4-dichlorophenoxyacetic acid (2,4-D), which is a widely used herbicide. Bioremediation offers an efficient, cost-effective and environmentally friendly method for degradation of the compound through the activities of the various microbial enzymes involved in the catabolic pathway. The aim of this study was to isolate and characterize bacterial isolate indigenous to contaminated sites in Durban, South Africa for 2,4-DCP degradation. One bacterium capable of utilizing 2,4-DCP as sole carbon source was isolated using culture enrichment technique and identified as Pseudomonas chlororaphis strain UFB2 via PCR amplification and analysis of 16S rRNA gene sequence. This isolate was able to degrade up to 75.11% of 2,4-DCP in batch cultures within 10 days, with the degradation rate constant of 0.14 mg/l/d. Phylogenetic analysis revealed the relatedness of this bacterial isolate to other Pseudomonas sp. previously characterized for chlorophenol degradation. PCR amplification of the catabolic genes involved in 2,4-DCP degradation revealed the presence of the correct amplicons for phenol hydroxylase (600 bp), catechol 1,2-dioxygenase (214 bp), muconate isomerase (851 bp), cis-dienelactone hydrolase (577 bp), and trans-dienelactone hydrolase (491 bp) genes. Enzyme assays revealed activity as high as 21840 mU/mg, 15630 mU/mg, 2340 mU/mg and 1490 mU/mg obtained for phenol hydroxylase, catechol 1,2-dioxygenase, cis-dienelactone hydroxylase and trans-dienelactone hydroxylase, respectively. The absence of catechol 2,3-dioxygenase gene and the corresponding enzyme in this isolate suggests that the organism followed ortho-pathway for 2,4-DCP degradation. Furthermore, the absence of malaycetate reductase genes showed that the bacterium may not be able to completely mineralize 2,4-DCP. Further studies are required to optimize 2,4-DCP degradation by this isolate as well as to elucidate the mechanism of 2,4-DCP degradation.

Keywords: biodegradation, catechol 1, 2-dioxygenase, 2, 4-dichlorophenol, phenol hydroxylase, Pseudomonas chlororaphis

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