Search results for: chaperone assisted protein folding

Authors: R. Asmeda, A. Noorlaila, M. H. Norziah

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This research work was conducted to investigate the effects of different grinding techniques during the milling process of rice grains on physicochemical characteristics of rice flour produced. Dry grinding, semi-wet grinding, and wet grinding were employed to produce the rice flour. The results indicated that different grinding methods significantly (p ≤ 0.05) affected physicochemical and functional properties of starch except for the carbohydrate content, x-ray diffraction pattern and breakdown viscosity. Dry grinding technique caused highest percentage of starch damage compared to semi-wet and wet grinding. Protein, fat and ash content were highest in rice flour obtained by dry grinding. It was found that wet grinding produce flour with smallest average particle size (8.52 µm), resulting in highest process yield (73.14%). Pasting profiles revealed that dry grinding produce rice flour with significantly lowest pasting temperature and highest setback viscosity.

Keywords: average particle size, grinding techniques, physicochemical characteristics, rice flour

Procedia PDF Downloads 196

Authors: Y. Okumuş, A. Tuna, A. T. Seyhan, H. Çelebi

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Useful carbon fibers were derived from chicken feathers (PCFs) based on a two-step pyrolysis method. The collected PCFs were cleaned and categorized as black, white and brown. Differential scanning calorimeter (DSC) and thermo-gravimetric analyzer (TGA) were systemically used to design the pyrolysis steps. Depending on colors, feathers exhibit different glass transition (Tg) temperatures. Long-time heat treatment applied to the feathers emerged influential on the surface quality of the resulting carbon fibers. Fourier Transformation Infrared (FTIR) examination revealed that the extent of disulfide bond cleavage is highly associated with the feather melting stability. Scanning electron microscopy (SEM) examinations were employed to evaluate the morphological changes of feathers after pyrolysis. Of all, brown feathers were found to be the most promising to turn into useful carbon fibers without any trace of melting and shape distortion when pyrolysis was carried out at 230°C for 24 hours and at 450°C for 1 hour.

Keywords: poultry chicken feather, keratin protein fiber, pyrolysis, high carbonaceous fibers

Procedia PDF Downloads 333

Authors: Ling-Lang Huang, Yih-Jer Wu

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Empathy is crucial for a patient-physician relationship and medical professionalism. Internet activity, gaming, or even addiction, have been more and more common among medical students. However, there’s been no report showing whether internet behavior has a substantial impact on empathy in medical students to our best knowledge. All year-2 medical students taking the optional course 'Narrative, Comprehension, and Communication' were enrolled. Internet behaviors are divided into two groups, 'internet users without online gaming (IU)' and 'internet users with online gaming (IG)', each group was further divided into 3 groups according to their average online retention time each day (< 2, 2 - 6, > 6 hours). Empathy was evaluated by the scores of the reports and humanities reflection after watching indicated movies, and by self-measured empathy questionnaire. All students taking the year-2 optional course 'Narrative, Comprehension, and Communication' were enrolled. As compared with students in the IU group, those in the IG group had significantly lower scores for the reports (81.3 ± 3.7 vs. 86.4 ± 5.1, P = 0.014). If further dividing the students into 5 groups (IU < 2, IU 2-6, IG < 2, IG 2 - 6, and IG > 6 hours), the scores were significantly and negatively correlated to online gaming with longer hours (r = -0.556, P = 0.006). However, there was no significant difference between IU and IG groups (33.0 ± 5.4 vs. 34.8 ± 3.2, P = n.s.), in terms of scores in the self-measured empathy questionnaire, neither was there any significant trend of scores along with longer online hours across the 5 groups (r = -0.164, P = n.s.). To date, there has been no evidence showing whether different internet behaviors (with or without online gaming) have distinct impacts on empathy. Although all of the medical students had a similarly good self-perception for empathy, our data suggested that online gaming did have a negative impact on their actual expression of empathy. Our observation has brought up an important issue for pondering: May IT- or gaming-assisted medical learning actually harm students’ empathy? In conclusion, this data suggests that long hours of online gaming harms expression of empathy, though all medics think themselves a person of high empathy.

Keywords: empathy. Internet, medical students, online gaming

Procedia PDF Downloads 135

Authors: Cristiano Giordani, Marco Diociaiuti, Cecilia Bombelli, Laura Zanetti-Polzi, Marcello Belfiore, Raoul Fioravanti, Gianfranco Macchia

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We investigated induced functional effects by evaluating Ca2+-influx in liposomes and cell viability in HT22-DIFF neurons. Only solutions rich in unstructured Prefibrillar-Oligomers (PFOs) were able, in the presence of Monosialoganglioside-GM1 (GM1), to induce Ca2+-influx and were also neurotoxic, suggesting a correlation between the two phenomena. Thus, in the presence of GM1, we investigated the protein conformation and liposome modification due to the interaction. Circular Dichroism showed that GM1 fostered the formation of β-structures and Energy Filtered-Transmission Electron Microscopy that PFOs formed “amyloid-channels” as reported for Aβ. We speculate that electrostatic forces occurring between the positive PFOs and negative GM1 drive the initial binding, while the hydrophobic profile of the flexible PFO is responsible for the subsequent pore formation. Conversely, the rigid β-structured mature/fibers (MFs) and proto-fibers (PFs) were unable to induce membrane damage and Ca2+- influx.

Keywords: amyloid proteins, neurotoxicity, lipid-rafts, GM1

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Authors: Yeng Min Yi, Rosli Md Illias, Salehhuddin Hamdan

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Industrial biocatalysis is mainly based on the use of cell free or intracellular enzyme systems. However, the expensive cost and relatively lower operational stability of free enzymes limit practical use in industries. Cell surface display system can be used as a cost-efficient alternative to overcome the laborious purification and substrate transport limitation. In this research, TibA autotransporter from E. coli was used to display Aspergillus fumigatus xylanase (xyn). The amplified xyn was fused in between N-terminal signal peptide and C-terminal β-barrel of TibA. The cloned was transformed and expressed in E. coli BL21 (DE3). Outer membrane localization of TibA-xyn fusion protein was confirmed by SDS PAGE and western blot with expected size of 62.5 kDa. Functional display of xyn was examined by activity assay. Cell surface displayed xyn exhibited the highest activity at 37 °c, 0.3 mM IPTG. As a summary, TibA displaying system has the potential for further industrial applications. Moreover, this is the first report of the display of xylanase using TibA on the surface of E. coli.

Keywords: biocatalysis, cell surface display, Escherichia coli, TibA autotransporter

Procedia PDF Downloads 286

Authors: Eirini Arvanitaki

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Romance has incrementally grown in popularity over the last century. The first Mills & Boon romance novel was published in 1909 and since then romance has not only survived but it has become a long standing genre. There are several reasons behind its durability and success. First, its ability to please and appeal to a mass audience. Romance novels are products of commercial success situated in large scale production, especially if one takes into account the high volume of romance novels published, translated and distributed all around the world every month. Second, what has also contributed to keeping the romance genre alive is the content of the books and their effect on the reader. These are stories of two heterosexual individuals who meet, fall in love, face obstacles and successfully overcome them. Through the love plots, the books address anxieties, concerns and everyday troubles that the average reader can identify with. Additionally, the romance novel is a means of escapism from everyday life and responsibilities as well as a short-lived opportunity to enjoy personal time/space and focus on one’s self. Third, the genre’s ability to adapt to the periods and societies in which it is published has also assisted in prolonging its longevity. This paper discusses the ways in which popular romance authors write and engage with the body. Despite the claim that popular romance narratives adjust their contents in accordance with different time periods and social phenomena, the paper highlights the dissimilarities between writing the female and male body and suggests that women romance writers are yet to break free from phallogocentric law. The examination of the projections of the body and the language used to describe it indicates that these narratives are flexible enough to adjust to twenty-first century but only within the limits of their own conventionality.

Keywords: body, gender, language, literary representations, popular romance narratives, taboo

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Authors: Manab Bandhu Bera, Navdeep Singh, Paramjit Singh Panesar

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In view of exploiting the health benefits of probiotic yeast S.boulardii NCDC 363 and make it available in the form of non-dairy food products, study was undertaken. In this, probiotic yeast S.boulardii NCDC 363 was incorporated in the edible film made from sodium alginate (SA), whey protein concentrate (WPC) and glycerol (50%). Response surface methodology was used to optimize process variables such as; concentration of SA (0.25-0.75%), WPC (1-2%) and temperature (70-80°C) and also to investigate effect of these process variables on viability of probiotic yeast and hardness when applied as an edible coat on extruded food products. Accelerated storage stability of optimized probiotic extruded food products samples was determined at 38 C and 90% RH. The optimized products were packed in high-density polyethylene (HDPE) and aluminum laminated polyethylene (ALP) pouches at 38°C and relative humidity maintained was 90%. It was observed that product stored in ALP had better stability in terms of moisture absorption, hardness and viability.

Keywords: probiotic yeast, extruded food product, WPC, RSM

Procedia PDF Downloads 278

Authors: Sandeep Kumar Agrawal, Aditi Bhattacharya, Janvie Manhas, Krushna Bhatt, Yatin Kholakiya, Nupur Khera, Ajoy Roychoudhury, Sudip Sen

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Autologous cultured explants of human oral mucosal epithelial cells (OMEC) are a potential therapeutic modality for limbal stem cell deficiency (LSCD). Injury or inflammation of the ocular surface in the form of burns, chemicals, Stevens Johnson syndrome, ocular cicatricial pemphigoid etc. can lead to destruction and deficiency of limbal stem cells. LSCD manifests in the form of severe ocular surface diseases (OSD) characterized by persistent and recurrent epithelial defects, conjuntivalisation and neovascularisation of the corneal surface, scarring and ultimately opacity and blindness. Most of the cases of OSD are associated with severe dry eye pertaining to diminished mucin and aqueous secretion. Mycophenolate mofetil (MMF) has been shown to upregulate the mucin expression in conjunctival goblet cells in vitro. The aim of this study was to evaluate the effects of MMF on mucin expression in primary cultures of oral mucosal epithelial cells. With institutional ethics committee approval and written informed consent, thirty oral mucosal epithelial tissue samples were obtained from patients undergoing oral surgery for non-malignant conditions. OMEC were grown on human amniotic membrane (HAM, obtained from expecting mothers undergoing elective caesarean section) scaffold for 2 weeks in growth media containing DMEM & Ham’s F12 (1:1) with 10% FBS and growth factors. In vitro dosage of MMF was standardised by MTT assay. Analysis of stem cell markers was done using RT-PCR while mucin mRNA expression was quantified using RT-PCR and q-PCR before and after treating cultured OMEC with graded concentrations of MMF for 24 hours. Protein expression was validated using immunocytochemistry. Morphological studies revealed a confluent sheet of proliferating, stratified oral mucosal epithelial cells growing over the surface of HAM scaffold. The presence of progenitor stem cell markers (p63, p75, β1-Integrin and ABCG2) and cell surface associated mucins (MUC1, MUC15 and MUC16) were elucidated by RT-PCR. The mucin mRNA expression was found to be upregulated in MMF treated primary cultures of OMEC, compared to untreated controls as quantified by q-PCR with β-actin as internal reference gene. Increased MUC1 protein expression was validated by immunocytochemistry on representative samples. Our findings conclude that OMEC have the ability to form a multi-layered confluent sheet on the surface of HAM similar to a cornea, which is important for the reconstruction of the damaged ocular surface. Cultured OMEC has stem cell properties as demonstrated by stem cell markers. MMF can be a novel enhancer of mucin production in OMEC. It has the potential to improve dry eye in patients undergoing OMEC transplantation for bilateral OSD. Further clinical trials are required to establish the role of MMF in patients undergoing OMEC transplantation.

Keywords: limbal stem cell deficiency, mycophenolate mofetil, mucin, ocular surface disease

Procedia PDF Downloads 336

Authors: Jiaqian Yin, Ruoling Chen, David Churchill, Huijuan Zou, Peipei Guo, Chunmei Liang, Xiaoqing Peng, Zhikang Zhang, Weiju Zhou, Yunxia Cao

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Purpose: This study aimed to explore the interaction of male and female age on implantation failure from in vitro fertilisation (IVF)/ intracytoplasmic sperm injection (ICSI) treatments in couples following their first cycles using the Anhui Maternal-Child Health Study (AMCHS). Methods: The AMCHS recruited 2042 infertile couples who were physically fit for in vitro fertilisation (IVF) or intracytoplasmic sperm injection (ICSI) treatment at the Reproductive Centre of the First Affiliated Hospital of Anhui Medical University between May 2017 to April 2021. This prospective cohort study analysed the data from 1910 cohort couples for the current paper data analysis. The multivariate logistic regression model was used to identify the effect of male and female age on implantation failure after controlling for confounding factors. Male age and female age were examined as continuous and categorical (male age: 20-<25, 25-<30, 30-<35, 35-<40, ≥40; female age: 20-<25, 25-<30, 30-<35, 35-<40, ≥40) predictors. Results: Logistic regression indicated that advanced maternal age was associated with increased implantation failure (P<0.001). There was evidence of an interaction between maternal age (30-<35 and ≥ 35) and paternal age (≥35) on implantation failure. (p<0.05). Only when the male was ≥35 years of increased maternal age was associated with the risk of implantation failure. Conclusion: In conclusion, there was an additive effect on implantation failure with advanced parental age. The impact of advanced maternal age was only seen in the older paternal age group. The delay of childbearing in both men and women will be a serious public issue that may contribute to a higher risk of implantation failure in patients needing assisted reproductive technology (ART).

Keywords: parental age, infertility, cohort study, IVF

Procedia PDF Downloads 159

Authors: Allison Herlene Du Plessis, Dalena (R.M.) Van Rooyen, Wilma Ten Ham-Baloyi, Sihaam Jardien-Baboo

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Introduction: Women die in pregnancy and childbirth for five main reasons—severe bleeding, infections, unsafe abortions, hypertensive disorders (pre-eclampsia and eclampsia), and medical complications including cardiac disease, diabetes, or HIV/AIDS complicated by pregnancy. In 2015, WHO classified sepsis as the third highest cause for maternal mortalities in the world. Chorioamnionitis is a clinical syndrome of intrauterine infection during any stage of the pregnancy and it refers to ascending bacteria from the vaginal canal up into the uterus, causing infection. While the incidence rates for chorioamnionitis are not well documented, complications related to chorioamnionitis are well documented and midwives still struggle to identify this condition in time due to its complex nature. Few diagnostic methods are available in public health services, due to escalated laboratory costs. Often the affordable biomarkers, such as C-reactive protein CRP, full blood count (FBC) and WBC, have low significance in diagnosing chorioamnionitis. A lack of screening impacts on effective and timeous management of chorioamnionitis, and early identification and management of risks could help to prevent neonatal complications and reduce the subsequent series of morbidities and healthcare costs of infants who are health foci of perinatal infections. Objective: This integrative literature review provides an overview of current best research evidence on the screening of women at risk for chorioamnionitis. Design: An integrative literature review was conducted using a systematic electronic literature search through EBSCOhost, Cochrane Online, Wiley Online, PubMed, Scopus and Google. Guidelines, research studies, and reports in English related to chorioamnionitis from 2008 up until 2020 were included in the study. Findings: After critical appraisal, 31 articles were included. More than one third (67%) of the literature included ranked on the three highest levels of evidence (Level I, II and III). Data extracted regarding screening for chorioamnionitis was synthesized into four themes, namely: screening by clinical signs and symptoms, screening by causative factors of chorioamnionitis, screening of obstetric history, and essential biomarkers to diagnose chorioamnionitis. Key conclusions: There are factors that can be used by midwives to identify women at risk for chorioamnionitis. However, there are a paucity of established sociological, epidemiological and behavioral factors to screen this population. Several biomarkers are available to diagnose chorioamnionitis. Increased Interleukin-6 in amniotic fluid is the better indicator and strongest predictor of histological chorioamnionitis, whereas the available rapid matrix-metalloproteinase-8 test requires further testing. Maternal white blood cells count (WBC) has shown poor selectivity and sensitivity, and C-reactive protein (CRP) thresholds varied among studies and are not ideal for conclusive diagnosis of subclinical chorioamnionitis. Implications for practice: Screening of women at risk for chorioamnionitis by health care providers providing care for pregnant women, including midwives, is important for diagnosis and management before complications arise, particularly in resource-constraint settings.

Keywords: chorioamnionitis, guidelines, best evidence, screening, diagnosis, pregnant women

Procedia PDF Downloads 128

Authors: Shady S. Hassan, Brijesh K. Tiwari, Gwilym A. Williams, Amit K. Jaiswal

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EU is known as the destination with the highest rate of the coffee consumption per capita in the world. Spent coffee grounds (SCG) are the main by-product of coffee brewing. SCG is either disposed as a solid waste or employed as compost, although the polysaccharides from such lignocellulosic biomass might be used as feedstock for fermentation processes. However, SCG as a lignocellulose have a complex structure and pretreatment process is required to facilitate an efficient enzymatic hydrolysis of carbohydrates. However, commonly used pretreatment methods, such as chemical, physico-chemical and biological techniques are still insufficient to meet optimal industrial production requirements in a sustainable way. Ultrasound is a promising candidate as a sustainable green pretreatment solution for lignocellulosic biomass utilization in a large scale biorefinery. Thus, ultrasound pretreatment of SCG without adding harsh chemicals investigated as a green technology to enhance enzyme hydrolysis. In the present work, ultrasound pretreatment experiments were conducted on SCG using different ultrasound frequencies (25, 35, 45, 130, and 950 kHz) for 60 min. Regardless of ultrasound power, low ultrasound frequency is more effective than high ultrasound frequency in pretreatment of biomass. Ultrasound pretreatment of SCG (at ultrasound frequency of 25 kHz for 60 min) followed by enzymatic hydrolysis resulted in total reducing sugars of 56.1 ± 2.8 mg/g of biomass. Fourier transform Infrared Spectroscopy (FTIR) was employed to investigate changes in functional groups of biomass after pretreatment, while high-performance liquid chromatography (HPLC) was employed for determination of glucose. Pretreatment of lignocellulose by low frequency ultrasound in water only was found to be an effective green approach for SCG to improve saccharification and glucose yield compared to native biomass. Pretreatment conditions will be optimized, and the enzyme hydrolysate will be used as media component substitute for the production of ethanol.

Keywords: lignocellulose, ultrasound, pretreatment, spent coffee grounds

Procedia PDF Downloads 329

Authors: R. S. Fisca, Y. R. Elox, L. Umi, U. Z. Luttfia, Kun Harismah

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Petai (Parkia speciosa) is a plant indigenous to Southeast Asia. It is consumed either raw or cooked. It has been used in folk medicine to treat diabetes, hypertension, and kidney problems. It contains minerals and vitamins. Petai contains a lot of chemical compounds that are beneficial for health, including antioxidants, Vitamin B6 0,9mg, energy 142 g. cal, 10.4 g protein. 2 g fat, 22 g carbohydrates, 95 mg calcium, phosphorus 115 mg, 1 mg iron, 200 IU of vitamin A, vitamin B1 0.17 mg, 36 mg of vitamin C that can resolve various health problems. These chips are the result of innovation from petai packaged in such a way becomes a tasty snack chips and can be enjoyed by many people to relax and also nutritious for health. In the manufacture of petai chips require several steps of them start by boiling, flating, drying and the last frying. In introducing the products widely we sell petai chips with several methods. Some of these methods include direct sales, delivery order, online/social media, and open some booth at a few places and the car free day in Solo every sunday. Opportunity in selling petai chips is very wide because there is no competitors with similar business. With the innovation of petai chips become healthy snacks can be introduced to the public and can even be exported out of the country as one of the extraordinary snacks from Indonesia.

Keywords: antioxidants, chips, healty, petai

Procedia PDF Downloads 568

Authors: Truong H. Le, Ngoc M. To, Quang N. Tran, Luu T. Cao, Chi V. Le

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Introduction: In Vietnam, the current monitoring and treatment for ordinary diabetic patient mostly based on glucose monitoring with HbA1c test for every three months (recommended goal is HbA1c < 6.5%~7%). For diabetes in pregnant women or Gestational diabetes mellitus (GDM), glycemic control until the time of delivery is extremly important because it could reduce significantly medical implications for both the mother and the child. Besides, GDM requires continuos glucose monitoring at least every two weeks and therefore an alternative marker of glycemia for short-term control is considering a potential tool for the healthcare providers. There are published studies have indicated that the glycosylated serum protein is a better indicator than glycosylated hemoglobin in GDM monitoring. Based on the actual practice in Vietnam, this study was designed to evaluate the role of serum fructosamine as a monitoring tool in GDM treament and its correlations with fasting blood glucose (G0), 2-hour postprandial glucose (G2) and glycosylated hemoglobin (HbA1c). Methods: A cohort study on pregnant women diagnosed with GDM by the 75-gram oralglucose tolerance test was conducted at Endocrinology Department, Cho Ray hospital, Vietnam from June 2014 to March 2015. Cho Ray hospital is the final destination for GDM patient in the southern of Vietnam, the study population has many sources from other pronvinces and therefore researchers belive that this demographic characteristic can help to provide the study result as a reflection for the whole area. In this study, diabetic patients received a continuos glucose monitoring method which consists of bi-weekly on-site visit every 2 weeks with glycosylated serum protein test, fasting blood glucose test and 2-hour postprandial glucose test; HbA1c test for every 3 months; and nutritious consultance for daily diet program. The subjects still received routine treatment at the hospital, with tight follow-up from their healthcare providers. Researchers recorded bi-weekly health conditions, serum fructosamine level and delivery outcome from the pregnant women, using Stata 13 programme for the analysis. Results: A total of 500 pregnant women was enrolled and follow-up in this study. Serum fructosamine level was found to have a light correlation with G0 ( r=0.3458, p < 0.001) and HbA1c ( r=0.3544, p < 0.001), and moderately correlated with G2 ( r=0.4379, p < 0.001). During study timeline, the delivery outcome of 287 women were recorded with the average age of 38.5 ± 1.5 weeks, 9% of them have macrosomia, 2.8% have premature birth before week 35th and 9.8% have premature birth before week 37th; 64.8% of cesarean section and none of them have perinatal or neonatal mortality. The study provides a reference interval of serum fructosamine for GDM patient was 112.9 ± 20.7 μmol/dL. Conclusion: The present results suggests that serum fructosamine is as effective as HbA1c as a reflection of blood glucose control in GDM patient, with a positive result in delivery outcome (0% perinatal or neonatal mortality). The reference value of serum fructosamine measurement provided a potential monitoring utility in GDM treatment for hospitals in Vietnam. Healthcare providers in Cho Ray hospital is considering to conduct more studies to test this reference as a target value in their GDM treatment and monitoring.

Keywords: gestational diabetes mellitus, monitoring tool, serum fructosamine, Vietnam

Procedia PDF Downloads 282

Authors: Linnan Zhang

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Linguists, as well as psychologists, have shown great interests in implicit causality in reference processing. However, most frequently-used approaches to this issue are psychological experiments (such as eye tracking or self-paced reading, etc.). This research is a corpus-based one and is assisted with statistical tool – software R. The main focus of the present study is about the competition between verb-based implicit causality and theme structure’s influence on anaphora bias in Mandarin Chinese sentences. In Accessibility Theory, it is believed that salience, which is also known as accessibility, and relevance are two important factors in reference processing. Theme structure, which is a special syntactic structure in Chinese, determines the salience of an antecedent on the syntactic level while verb-based implicit causality is a key factor to the relevance between antecedent and anaphora. Therefore, it is a study about anaphora, combining psychology with linguistics. With analysis of the sentences from corpus as well as the statistical analysis of Multinomial Logistic Regression, major findings of the present study are as follows: 1. When the sentence is stated in a ‘cause-effect’ structure, the theme structure will always be the antecedent no matter forward biased verbs or backward biased verbs co-occur; in non-theme structure, the anaphora bias will tend to be the opposite of the verb bias; 2. When the sentence is stated in a ‘effect-cause’ structure, theme structure will not always be the antecedent and the influence of verb-based implicit causality will outweigh that of theme structure; moreover, the anaphora bias will be the same with the bias of verbs. All the results indicate that implicit causality functions conditionally and the noun in theme structure will not be the high-salience antecedent under any circumstances.

Keywords: accessibility theory, anaphora, theme strcture, verb-based implicit causality

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Authors: Threera Intarareaung

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This research aims to study guidelines of organizing the Wat Sai Museum of Local Boats, of Ruam Sai Pattana Community 789, Kradangnga sub-district, Bang Khon Tee district, Samut Songkram province. The procedures are to seek for potentials and problems within the community to properly organize the said museum. The research contains methodology of qualitative by studying from the population from Ruam Sai Pattana 789, a community that located within the area of the museum. The sample contains 40 persons. The procedures are in-depth interview, observation, focus group by discussing ideas, problems and the importance of the museum. After that, the information received is synthesized and analyzed. The result finds that the local temple requires setting up the boat museum. However, there are several factors that limit the possibility which are the museum manager, resources, exhibition and service and knowledge management. These factors reflect mutual potentials and problems that lead us to seek the best solutions to set up the local boat museum. In conclusion, there are 4 issues regarding the management. First, the knowledge management should be conducted to indicate the purposes of the museum. Second, exhibition and service should be categorized. In addition, the serviced should be assisted by local guides. Third, the museum should employ staffs to correspond to other entities such as related authorities. Fourth is regarding location and condition of the building. This is considered as a major factor that needs discussion from different parties. However, the condition of the existing resource is proper. All the information is collected by field trip and interview the locals and person of interest. Therefore, the idea is inconsistent with the community’s requirement that can proceed to real practice.

Keywords: museum, boat museum, heritage tourism, conservation lifestyle

Procedia PDF Downloads 335

Authors: Srie Rezeki Nur Endah, Richa Mardianingrum

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Cancer is one of the most feared diseases and is considered the leading cause of death worldwide. Generally, cancer drugs are synthetic drugs with relatively more expensive prices and have harmful side effects, so many people turn to traditional medicine, for example by utilizing herbal medicine. Erythrina poeppigiana is one of the plants that can be used as a medicinal plant containing 10,11-dihidroerisodin compounds that are useful anticancer etnofarmakologi. The purpose of this study was to identify the target of 10,11 dihydroerisodin receptor compound as in silico anticancer candidate. The pure isolate was tested physicochemically by MS (Mass Spectrometry), UV-Vis (Ultraviolet – Visible), IR (Infra Red), 13C-NMR (Carbon-13 Nuclear Magnetic Resonance), 1H-NMR (Hydrogen-1 Nuclear Magnetic Resonance), to obtain the structure of 10,11-dihydroerisodin alkaloid compound then identified to target receptors in silico. From the results of the study, it was found that 10,11-dihydroerisodin compound can work on the Serine / threonine-protein kinase Chk1 receptor that serves as an anti-cancer candidate.

Keywords: anti-cancer, Erythrina poeppigiana, target receptor, 10, 11- dihidroerisodin

Procedia PDF Downloads 247

Authors: Antonela Matana, Dubravka Brdar, Vesela Torlak, Marijana Popovic, Ivana Gunjaca, Ozren Polasek, Vesna Boraska Perica, Maja Barbalic, Ante Punda, Caroline Hayward, Tatijana Zemunik

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Parathyroid hormone (PTH) plays a critical role in the regulation of bone mineral metabolism and calcium homeostasis. Higher PTH levels are associated with heart failure, hypertension, coronary artery disease, cardiovascular mortality and poorer bone health. A twin study estimated that 60% of the variation in PTH concentrations is genetically determined. Only one GWAS of PTH concentration has been reported to date. Identified loci explained 4.5% of the variance in circulating PTH, suggesting that additional genetic variants remain undiscovered. Therefore, the aim of this study was to identify novel genetic variants associated with PTH levels in a general population. We have performed a GWAS meta-analysis on 2596 individuals originating from three Croatian cohorts: City of Split and the Islands of Korčula and Vis, within a large-scale project of “10,001 Dalmatians”. A total of 7 411 206 variants, imputed using the 1000 Genomes reference panel, with minor allele frequency ≥ 1% and Rsq ≥ 0.5 were analyzed for the association. GWAS within each data set was performed under an additive model, controlling for age, gender and relatedness. Meta-analysis was conducted using the inverse-variance fixed-effects method. Furthermore, to identify sex-specific effects, we have conducted GWAS meta-analyses analyzing males and females separately. In addition, we have performed biological pathway analysis. Four SNPs, representing one locus, reached genome-wide significance. The most significant SNP was rs11099476 on chromosome 4 (P=1.15x10-8), which explained 1.14 % of the variance in PTH. The SNP is located near the protein-coding gene RASGEF1B. Additionally, we detected suggestive association with SNPs, rs77178854 located on chromosome 2 in the DPP10 gene (P=2.46x10-7) and rs481121 located on chromosome 1 (P=3.58x10-7) near the GRIK1 gene. One of the top hits detected in the main meta-analysis, intron variant rs77178854 located within DPP10 gene, reached genome-wide significance in females (P=2.21x10-9). No single locus was identified in the meta-analysis in males. Fifteen biological pathways were functionally enriched at a P<0.01, including muscle contraction, ion homeostasis and cardiac conduction as the most significant pathways. RASGEF1B is the guanine nucleotide exchange factor, known to be associated with height, bone density, and hip. DPP10 encodes a membrane protein that is a member of the serine proteases family, which binds specific voltage-gated potassium channels and alters their expression and biophysical properties. In conclusion, we identified 2 novel loci associated with PTH levels in a general population, providing us with further insights into the genetics of this complex trait.

Keywords: general population, genome-wide association analysis, parathyroid hormone, single nucleotide polymorphisms.

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Authors: Sung-Hee Hwang, Michael Lee

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Upregulated Src activity has been implicated in a variety of cancers. Thus, Src family tyrosine kinase (SFK) inhibitors are often effective cancer treatments. Here, we investigate the role of autophagy in ATG5 knockout cell lines generated by the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas mediated genome editing. The CRISPR-associated protein Cas9 is an RNA-guided DNA endonuclease that uses RNA–DNA complementarity to identify target sites for sequence specific double-stranded DNA (dsDNA) cleavage. Interestingly, ATG5 KO cells clearly showed a greater proliferation rate than WT NIH 3T3 cells, implying that autophagy induction is cytotoxic. Also, the clonogenic survival of ATG5 KO cells was greater than WT cells. The MTT assay revealed that the cytotoxic effect of PP2 was weaker on ATG5 knockout cells than that WT cells. The conversion of non-autophagic LC3-I to autophagic LC3-II and RT-PCR confirmed the functional gene knockout. Furthermore, Cyto-ID autophagy assay also revealed that PP2 failed to induce autophagy in ATG5 knockout cells. Together, our findings suggest that the resistance to PP2 in ATG5 knockout cells is associated with defective autophagy.

Keywords: ATG5 knockout, Autophagy, CRISPR/Cas9, PP2

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Authors: T. Büyüksırıt, H. Kuleaşan

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Natural antimicrobials are used to preserve foods that can be found in plants, animals, and microorganisms. Antimicrobial substances are natural or artificial agents that produced by microorganisms or obtained semi/total chemical synthesis are used at low concentrations to inhibit the growth of other microorganisms. Food borne pathogens and spoilage microorganisms are inactivated by the use of antagonistic microorganisms and their metabolites. Yeasts can produce toxic proteins or glycoproteins (toxins) that cause inhibition of sensitive bacteria and yeast species. Antimicrobial substance producing phenotypes belonging different yeast genus were isolated from different sources. Toxins secreted by many yeast strains inhibiting the growth of other yeast strains. These strains show antimicrobial activity, inhibiting the growth of mold and bacteria. The effect of antimicrobial agents produced by yeasts can be extremely fast, and therefore may be used in various treatment procedures. Rapid inhibition of microorganisms is possibly caused by microbial cell membrane lipopolysaccharide binding and in activation (neutralization) effect. Antimicrobial agents inhibit the target cells via different mechanisms of action.

Keywords: antimicrobial agents, yeast, toxic protein, glycoprotein

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Authors: A. K. Tursunova, O. V. Chebonenko, A. Zh. Amirkulova, A. O. Abaildayev, O. A. Sapko, Y. M. Dyo, A. Sh. Utarbaeva

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Fusarium solani and its variants cause root and stem rot of plants. Dry rot is the most common disease of potato tubers during storage. The causative agents of fusariosis in contact with plants behave as antagonists, growth stimulants or parasites. The diversity of host-parasite relationships is explained by the parasite’s ability to produce a wide spectrum of biologically active compounds including toxins, enzymes, oligosaccharides, antibiotic substances, enniatins and gibberellins. Many of these metabolites contribute to the creation of compatible relations; others behave as elicitors, inducing various protective responses in plants. An important part of the strategy for developing plant resistance against pathogens is the activation of protein synthesis to produce protective ‘pathogenesis-related’ proteins. The family of PR-proteins known to confer the most protective response is chitinases (EC 3.2.1.14, Cht) and β-1,3-glucanases (EC 3.2.1.39, Glu). PR-proteins also include a large multigene family of peroxidases (EC 1.11.1.7, Pod), and increased activity of Pod and expression of the Pod genes leads to the development of resistance to a broad class of pathogens. Despite intensive research on the role of PR-proteins, the question of their participation in the mechanisms of formation of the F.solani–S.tuberosum pathosуstem is not sufficiently studied. Our aim was to investigate the effect of different classes of F. solani metabolites on the activity of chitinase, β-1,3-glucanases and peroxidases in tubers of Solanum tuberosum. Metabolite culture filtrate (CF) and cytoplasmic components were fractionated by extraction of the mycelium with organic solvents, salting out techniques, dialysis, column chromatography and ultrafiltration. Protein, lipid, carbohydrate and polyphenolic fractions of fungal metabolites were derived. Using enzymatic hydrolysis we obtained oligo glycans from fungal cell walls with different molecular weights. The activity of the metabolites was tested using potato tuber discs (d = 16mm, h = 5mm). The activity of PR-proteins of tubers was analyzed in a time course of 2–24 hours. The involvement of the analysed metabolites in the modulation of both early non-specific and late related to pathogenesis reactions was demonstrated. The most effective inducer was isolated from the CF (fraction of total phenolic compounds including naphtazarins). Induction of PR-activity by this fraction was: chitinase - 340-360%, glucanase - 435-450%, soluble forms of peroxidase - 400-560%, related forms of peroxidase - 215-237%. High-inducing activity was observed by the chloroform and acetonitrile extracts of the mycelium (induction of chitinase and glucanase activity was 176-240%, of soluble and bound forms of peroxidase - 190-400%). The fraction of oligo glycans mycelium cell walls of 1.2 kDa induced chitinase and β-1,3-glucanase to 239-320%; soluble forms and related peroxidase to 198-426%. Oligo glycans cell walls of 5-10 kDa had a weak suppressor effect - chitinase (21-25%) and glucanase (25-28%) activity; had no effect on soluble forms of peroxidase, but induced to 250-270% activity related forms. The CF polysaccharides of 8.5 kDa and 3.1 kDa inhibited synchronously the glucanase and chitinase specific response in step (after 24 hours at 42-50%) and the step response induced nonspecific peroxidase activity: soluble forms 4.8 -5.2 times, associated forms 1.4-1.6 times.

Keywords: fusarium solani, PR-proteins, peroxidase, solanum tuberosum

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Authors: Ali Mohagheghzadeh, Samad Vaez Badiegard, Bita Shomali

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Nowadays, with the increase in population, the need for protein sources is increasing. Different bacteria can cause food poisoning while most of the symptoms of food poisoning are similar to those of gastrointestinal infections. As a result, the diagnosis of bacteria and viruses causing food poisoning would not be possible without a stool culture. Cases of food poisoning are often accompanied by gastrointestinal disorders such as diarrhea, vomit, and gastrointestinal stomach cramps. Thus, providing enough food, taking into account health issues has always been a concern of authorities. Since E. coli bacterium is one of the important indicators of food hygiene and quality, producing food without being contaminated by this bacterium is desired in the food industry. This study aimed at assessing the E. coli contamination of poultry meat produced in slaughterhouses. Samples were taken from critical areas of slaughterhouses, namely the feather picking area, viscera and carcass evacuation area the area after cooling chillers. The results showed that 60% of contamination occurs in feather picking area. Among antiseptic and detergent materials, the highest reduction belongs to Epimax.

Keywords: slaughterhouse, E. coli, Epimax, contamination

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Authors: Maryam Hamlehdar, Guillermo A. Narsilio

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To enhance the feasibility of utilising geothermal hot sedimentary aquifers (HSAs) for clean hydrogen production, one approach is the implementation of solar-integrated geothermal energy systems. This detailed modelling study conducts a thermo-economic assessment of an advanced Organic Rankine Cycle (ORC)-based hydrogen production system that uses low-temperature geothermal reservoirs, with a specific focus on hot sedimentary aquifers (HSAs) over a 30-year period. In the proposed hybrid system, solar-thermal energy is used to raise the water temperature extracted from the geothermal production well. This temperature increase leads to a higher steam output, powering the turbine and subsequently enhancing the electricity output for running the electrolyser. Thermodynamic modeling of a parabolic trough solar (PTS) collector is developed and integrated with modeling for a geothermal-based configuration. This configuration includes a closed regenerator cycle (CRC), proton exchange membrane (PEM) electrolyser, and thermoelectric generator (TEG). Following this, the study investigates the impact of solar energy use on the temperature enhancement of the geothermal reservoir. It assesses the resulting consequences on the lifecycle performance of the hydrogen production system in comparison with a standalone geothermal system. The results indicate that, with the appropriate solar collector area, a combined solar-geothermal hydrogen production system outperforms a standalone geothermal system in both cost and rate of production. These findings underscore a solar-assisted geothermal hybrid system holds the potential to generate lower-cost hydrogen with enhanced efficiency, thereby boosting the appeal of numerous low to medium-temperature geothermal sources for hydrogen production.

Keywords: clean hydrogen production, integrated solar-geothermal, low-temperature geothermal energy, numerical modelling

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Authors: Lea Stein, Sabine Rösner, Alessandra Lo Giudice, Beate Ditzen, Tewes Wischmann

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For many couples bearing children is a consistent life goal; however, it cannot always be fulfilled. Undergoing infertility treatment does not guarantee pregnancies and live births. Couples have to deal with miscarriages and sometimes even discontinue infertility treatment. Significant medical predictors for the outcome of infertility treatment have yet to be fully identified. To further our understanding, a cross-sectional five-year follow-up survey was undertaken, in which 95 women and 82 men that have been treated at the Women’s Hospital of Heidelberg University participated. Binary logistic regressions, parametric and non-parametric methods were used for our sample to determine the relevance of biological (infertility diagnoses, maternal and paternal age) and lifestyle factors (smoking, drinking, over- and underweight) on the outcome of infertility treatment (clinical pregnancy, live birth, miscarriage, dropout rate). During infertility treatment, 72.6% of couples became pregnant and 69.5% were able to give birth. Suffering from miscarriages 27.5% of couples and 20.5% decided to discontinue an unsuccessful fertility treatment. The binary logistic regression models for clinical pregnancies, live births and dropouts were statistically significant for the maternal age, whereas the paternal age in addition to maternal and paternal BMI, smoking, infertility diagnoses and infections, showed no significant predicting effect on any of the outcome variables. The results confirm an effect of maternal age on infertility treatment, whereas the relevance of other medical predictors remains unclear. Further investigations should be considered to increase our knowledge of medical predictors.

Keywords: advanced maternal age, assisted reproductive technology, female factor, male factor, medical predictors, infertility treatment, reproductive medicine

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Authors: Mbadugha Esther Ifeoma

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Soybeans, like other legumes are rich in nutrients. However, the nutrient profile of soybeans differs in some important ways from most other legumes. Among other nutrients, soy is high in protein, carbohydrates, and fibers, is rich in vitamins, minerals and unsaturated fatty acids and is low in saturated fatty acids. Because of its high nutritional value, it has been rated to be equivalent to meats, eggs and milk. Soy has many health benefits including prevention of coronary heart disease, prevention of cancer growth, improvement of cognitive function, promotion of bone health, prevention of obesity, prevention of type II diabetes and promotion of growth of normal floras in the colon. Soybean consumption is also associated with some side effects which include allergy, flatulence and abdominal discomfort. Nurses/health care providers should therefore, educate clients on the precautionary measures to be taken in preparing soy food products in order to reduce to the barest minimum the side effects, while encouraging them to include soy as part of their daily meals for optimal health and vitality.

Keywords: health benefit, health education, nutritional benefit, soybeans

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Authors: Ewelina Wisnik, Zsolt Regdon, Kinga Chmielewska, Laszlo Virag, Agnieszka Robaszkiewicz

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Apart from protecting genome, PARP1 has been documented to regulate many intracellular processes inter alia gene transcription by physically interacting with chromatin bound proteins and by their ADP-ribosylation. Our recent findings indicate that expression of PARP1 decreases during the differentiation of human CD34+ hematopoietic stem cells to monocytes as a consequence of differentiation-associated cell growth arrest and formation of E2F4-RBL2-HDAC1-SWI/SNF repressive complex at the promoter of this gene. Since the RBL2 complexes repress genes in a E2F-dependent manner and are widespread in the genome in G0 arrested cells, we asked (a) if RBL2 directly contributes to defining monocyte phenotype and function by targeting gene promoters and (b) if RBL2 controls gene transcription indirectly by repressing PARP1. For identification of genes controlled by RBL2 and/or PARP1,we used primer libraries for surface receptors and TLR signaling mediators, genes were silenced by siRNA or shRNA, analysis of gene promoter occupation by selected proteins was carried out by ChIP-qPCR, while statistical analysis in GraphPad Prism 5 and STATISTICA, ChIP-Seq data were analysed in Galaxy 2.5.0.0. On the list of 28 genes regulated by RBL2, we identified only four solely repressed by RBL2-E2F4-HDAC1-BRM complex. Surprisingly, 24 out of 28 emerged genes controlled by RBL2 were co-regulated by PARP1 in six different manners. In one mode of RBL2/PARP1 co-operation, represented by MAP2K6 and MAPK3, PARP1 was found to associate with gene promoters upon RBL2 silencing, which was previously shown to restore PARP1 expression in monocytes. PARP1 effect on gene transcription was observed only in the presence of active EP300, which acetylated gene promoters and activated transcription. Further analysis revealed that PARP1 binding to MA2K6 and MAPK3 promoters enabled recruitment of EP300 in monocytes, while in proliferating cancer cell lines, which actively transcribe PARP1, this protein maintained EP300 at the promoters of MA2K6 and MAPK3. Genome-wide analysis revealed a similar distribution of PARP1 and EP300 around transcription start sites and the co-occupancy of some gene promoters by PARP1 and EP300 in cancer cells. Here, we described a new RBL2/PARP1/EP300 axis which controls gene transcription regardless of the cell type. In this model cell, cycle-dependent transcription of PARP1 regulates expression of some genes repressed by RBL2 upon cell cycle limitation. Thus, RBL2 may indirectly regulate transcription of some genes by controlling the expression of EP300-recruiting PARP1. Acknowledgement: This work was financed by Polish National Science Centre grants nr DEC-2013/11/D/NZ2/00033 and DEC-2015/19/N/NZ2/01735. L.V. is funded by the National Research, Development and Innovation Office grants GINOP-2.3.2-15-2016-00020 TUMORDNS, GINOP-2.3.2-15-2016-00048-STAYALIVE and OTKA K112336. AR is supported by Polish Ministry of Science and Higher Education 776/STYP/11/2016.

Keywords: retinoblastoma transcriptional co-repressor like 2 (RBL2), poly(ADP-ribose) polymerase 1 (PARP1), E1A binding protein p300 (EP300), monocytes

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Authors: Maryam Borjali

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Introduction. Mortality due to noncommunicable diseases has increased in the world today with the advent of demographic shifts, growing age, and lifestyle patterns in the world, which have been affected by economic and social crises. Congenital heart defects are one of the forms of diseases that have raised infant mortality worldwide. e objective of present study was to identify nonmedical determinants related to this abnormality from the mother’s perspectives. Methods. is research was a qualitative study and the data collection method was a semistructured interview with mothers who had children with congenital heart diseases referring to the Shahid Rajaei Heart Hospital in Tehran, Iran. A thematic analysis approach was employed to analyze transcribed documents assisted by MAXQDA Plus version 12. Results. Four general themes and ten subthemes including social contexts (social harms, social interactions, and social necessities), psychological contexts (mood disorders and mental well-being), cultural contexts (unhealthy lifestyle, family culture, and poor parental health behaviors), and environmental contexts (living area and polluted air) were extracted from interviews with mothers of children with congenital heart diseases. Conclusions. Results suggest that factors such as childhood poverty, lack of parental awareness of congenital diseases, lack of proper nutrition and health facilities, education, and lack of medical supervision during pregnancy were most related with the birth of children with congenital heart disease from mothers’ prospective. In this regard, targeted and intersectorial collaborations are proposed to address nonmedical determinants related to the incidence of congenital heart diseases.

Keywords: congenital_cou, cultural, social, platform

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Authors: Supatra Karnjanapratum, Soottawat Benjakul

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Gelatin hydrolysate, especially from marine resource, has been known to possess antioxidative activity. Nevertheless, the activity is still lower in comparison with the commercially available antioxidant. Maillard reactions can be use to increase antioxidative activity of gelatin hydrolysate, in which the numerous amino group could be involved in glycation. In the present study, gelatin hydrolysate (GH) from unicorn leatherjacket skin prepared using glycyl endopeptidase with prior autolysis assisted process was used for preparation of Maillard reaction products (MRPs) under dry condition. The impacts of different factors including, types of saccharides, GH to saccharide ratio, incubation temperatures, relative humidity (RH) and times on antioxidative activity of MRPs were investigated. MRPs prepared using the mixture of GH and galactose showed the highest antioxidative activity as determined by both ABTS radical scavenging activity and ferric reducing antioxidant power during heating (0-48 h) at 60 °C with 65% RH, compared with those derived from other saccharide tested. GH to galactose ratio at 2:1 (w/w) yielded the MRPs with the highest antioxidative activity, followed by the ratios of 1:1 and 1:2, respectively. When the effects of incubation temperatures (50, 60, 70 °C) and RH (55, 65, 75%) were examined, the highest browning index and the absorbance at 280 nm were found at 70 °C, regardless of RH. The pH and free amino group content of MRPs were decreased with the concomitant increase in antioxidative activity as the reaction time increased. Antioxidative activity of MRPs generally increased with increasing temperature and the highest antioxidative activity was found when RH of 55% was used. Based on electrophoresis of MRP, the polymerization along with the formation of high molecular weight material was observed. The optimal condition for preparing antioxidative MRPs was heating the mixture of GH and galactose (2:1) at 70 °C and 55% RH for 36 h. Therefore, antioxidative activity of GH was improved by Maillard reaction and the resulting MRP could be used as natural antioxidant in food products.

Keywords: antioxidative activity, gelatin hydrolysate, maillard reaction, unicorn leatherjacket

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Authors: Helan Satish, M. Ramasubba Reddy

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The simulation of an endocardial cell for gene mutation in the cardiac sodium ion channel NaV1.5, encoded by SCN5A gene, is discussed. The characterization of Brugada Syndrome by loss of function effect on SCN5A mutation due to L812Q mutant present in the DII-S4 transmembrane region of the NaV1.5 channel protein and its effect in an endocardial cell is studied. Ten Tusscher model of human ventricular action potential is modified to incorporate the changes contributed by L812Q mutant in the endocardial cells. Results show that BrS-associated SCN5A mutation causes reduction in the inward sodium current by modifications in the channel gating dynamics such as delayed activation, enhanced inactivation, and slowed recovery from inactivation in the endocardial cell. A decrease in the inward sodium current was also observed, which affects depolarization phase (Phase 0) that leads to reduction in the spike amplitude of the cardiac action potential.

Keywords: SCN5A gene mutation, sodium channel, Brugada syndrome, cardiac arrhythmia, action potential

Procedia PDF Downloads 129

Authors: A.Knox

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Student diversity in postgraduate classes puts major challenges on educators seeking to encourage student engagement and desired to learn outcomes. This paper outlines the impact of a set of teaching initiatives aimed at addressing challenges associated with teaching and learning in an environment characterized by diversity in the student cohort. The study examines postgraduate students completing the core capstone unit within a specialized business degree. Although relatively small, the student cohort is highly diverse in terms of cultural backgrounds represented, prior learning and/or qualifications, as well as duration and type of work experience relevant to the degree, is completed. The wide range of cultures, existing knowledge and experience create enormous challenges with respect to students’ learning needs and outcomes. Subsequently, a suite of teaching innovations has been adopted to enhance curriculum content/delivery and the design of assessments. This paper explores the impact of these specific teaching and learning practices, examining the ways they have supported students’ diverse needs and enhanced students’ learning outcomes. Data from surveys and focus groups are used to assess the effectiveness of these practices. The results highlight the effectiveness of peer-assisted learning, cultural competence-building, and advanced assessment options in addressing diverse student needs and enhancing student engagement and learning outcomes. These findings suggest that such practices would benefit students’ learning in environments marked by diversity in the student cohort. Specific recommendations are offered for other educators working with diverse classes.

Keywords: assessment design, curriculum content, curriculum delivery, student diversity

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Authors: Farrah Putri Salmanida, Mei-Li Wu, Rika Wahyuningtyas, Wen-Bin Chung, Hso-Chi Chaung, Ko-Tung Chang

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Within the field of immunotherapy, oncolytic virotherapy (OVT) employs dual approaches that directly eliminate tumor cells while preserving healthy ones and indirectly reprogram the tumor microenvironment (TME) to elicit antitumor responses. Within the TME, tumor associated macrophages (TAMs) manifest characteristics akin to those of anti-inflammatory M2 macrophages, thus earning the designation of M2-like TAMs. In prior research, two antigens denoted as A1 (g6Ld10T) and A3 (ORF6L5), derived from a complete sequence of ORF5 with partial sequence of ORF6 in Porcine Reproductive and Respiratory Syndrome Virus Genotype 2 (PRRSV-2), demonstrated the capacity to repolarize M2-type porcine alveolar macrophages (PAMs) into M1 phenotypes. In this study, we sought for utilizing OVT strategies by introducing A1 or A3 on TAMs to endow them with the anti-tumor traits of M1 macrophages while retaining their capacity to target cancer cells. Upon exposing human THP-1-derived M2 macrophages to a cross-species test with 2 µg/ml of either A1 or A3 for 24 hours, real time PCR revealed that A3, but not A1, treated cells exhibited upregulated gene expressions of M1 markers (CCR7, IL-1ß, CCL2, Cox2, CD80). These cells reacted to virus-derived antigen, as evidenced by increased expression of pattern-recognition receptors TLR3, TLR7, and TLR9, subsequently providing feedback in the form of type I interferon responses like IFNAR1, IFN-ß, IRF3, IRF7, OAS1, Mx1, and ISG15. Through an MTT assay, only after 15 µg/ml of A3 treatment could the cell viability decrease, with a predicted IC50 of 16.96 µg/ml. Interestingly, A3 caused dose-dependent toxicity to a rat C6 glial cancer cell line even at doses as low as 2.5 µg/ml and reached its IC50 at 9.419 µg/ml. Using Annexin V/7AAD staining and PCR test, we deduced that a significant proportion of C6 cells were undergoing the early apoptosis phase predominantly through the intrinsic apoptosis cascade involving Bcl-2 family proteins. Following this stage, we conducted a test on A3’s repolarization ability, which revealed a significant rise in M1 gene expression markers, such as TNF, CD80, and IL-1ß, in M2-like TAMs generated in vitro from murine RAW264.7 macrophages grown with conditioned medium of 4T1 breast cancer cells. This was corroborated by the results of transcriptome analysis, which revealed that the primary subset among the top 10 to top 30 significantly upregulated differentially expressed genes (DEGs) dominantly consisted of M1 macrophages profiles, including Ccl3, Ccl4, Csf3, TNF, Bcl6b, Stc1, and Dusp2. Our findings unveiled the remarkable potential of the PRRSV-derived antigen A3 to repolarize macrophages while also being capable of selectively inducing apoptosis in cancerous cells. While further in vivo study is needed for A3, it holds promise as an adjuvant by its dual effects in cancer therapy modalities.

Keywords: cancer cell apoptosis, interferon responses, macrophage repolarization, recombinant protein

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