Search results for: differentially expressed gene
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2546

Search results for: differentially expressed gene

626 Using Wearable Technology to Monitor Perinatal Health: Perspectives of Community Health Workers and Potential Use by Underserved Perinatal Women in California

Authors: Tamara Jimah, Priscilla Kehoe, Pamela Pimentel, Amir Rahmani, Nikil Dutt, Yuqing Guo

Abstract:

Ensuring equitable access to maternal health care is critical for public health. Particularly for underserved women, community health workers (CHWs) have been invaluable in providing support through health education and strategies for improved maternal self-care management. Our research aimed to assess the acceptance of technology by CHWs and perinatal women to promote healthy pregnancy and postpartum wellness. This pilot study was conducted at a local community organization in Orange County, California, where CHWs play an important role in supporting low-income women through home visitations. Questionnaires were administered to 14 CHWs and 114 pregnant and postpartum women, literate in English and/or Spanish. CHWs tested two wearable devices (Galaxy watch and Oura ring) and shared their user experience, including potential reception by the perinatal women they served. In addition, perinatal women provided information on access to a smart phone and the internet, as well as their interest in using wearable devices to self-monitor personal health with guidance from a CHW. Over 85% of CHWs agreed that it was useful to track pregnancy with the smart watch and ring. The majority of perinatal women owned a smartphone (97.4%), had access to the internet (80%) and unlimited data plans (78%), expressed interest in using the smart wearable devices to self-monitor health, and were open to receiving guidance from a CHW (87%). Community health workers and perinatal women embraced the use of wearable technology to monitor maternal health. These preliminary findings have formed the basis of an ongoing research study that integrates CHW guidance and technology (i.e., smart watch, smart ring, and a mobile phone app) to promote self-efficacy and self-management among underserved perinatal women.

Keywords: community health workers, health promotion and education, health equity, maternal and child health, technology

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625 Beak Size and Asynchronous Hatch in Broiler Chicks

Authors: Mariana Thimotheo, Gabriel Carvalho Ripamonte, Marina De Almeida Nogueira, Silvia Camila Da Costa Aguiar, Marcelo Henrique Santana Ulian, Euclides Braga Malheiros, Isabel Cristina Boleli

Abstract:

Beak plays a fundamental role in the hatching process of the chicks, since it is used for internal and external pipping. The present study examined whether the size of the beak influences the birth period of the broiler chicks in the hatching window. It was analyzed the beak size (length, height and width) of one-hundred twenty nine newly hatched chicks from light eggs (56.22-61.05g) and one-hundred twenty six chicks from heavy eggs (64.95-70.90g), produced by 38 and 45 weeks old broiler breeders (Cobb 500®), respectively. Egg incubation occurred at 37.5°C and 60% RH, with egg turning every hour. Length, height and width of the beaks were measured using a digital caliper (Zaas precision - digital caliper 6", 0.01mm) and the data expressed in millimeters. The beak length corresponded to distance between the tip of the beak and the rictus. The height of the beak was measured in the region of the culmen and its width in the region of the nostrils. Data were analyzed following a 3x2 factorial experimental design, being three birth periods within the hatching window (early: 471.78 to 485.42h, intermediate: 485.43 to 512.27h, and late: 512.28 to 528.72h) and two egg weights (light and heavy). There was a significant interaction between birth period and egg weight for beak height (P < 0.05), which was higher in the intermediate chicks from heavy eggs than in the other chicks from the same egg weight and chicks from light eggs (P < 0.05), that did not differ (P > 0.05). The beak length was influenced only for a birth period, and decreased through the hatch window (early < intermediate < late) (P < 0.05). The width of the beaks was influenced by both main factors, birth period and egg weight (P < 0.05). Early and intermediate chicks had similar beak width, but greater than late chicks, and chicks from heavy eggs presented greater beak width than chicks from light eggs (P < 0.05). In sum, the results show that chicks with longer beak hatch first and that beak length is an important variable for hatch period determination mainly for light eggs.

Keywords: beak dimensions, egg weight, hatching period, hatching window

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624 Large-Scale Screening for Membrane Protein Interactions Involved in Platelet-Monocyte Interactions

Authors: Yi Sun, George Ed Rainger, Steve P. Watson

Abstract:

Background: Beyond the classical roles in haemostasis and thrombosis, platelets are important in the initiation and development of various thrombo-inflammatory diseases. In atherosclerosis and deep vein thrombosis, for example, platelets bridge monocytes with endothelium and form heterotypic aggregates with monocytes in the circulation. This can alter monocyte phenotype by inducing their activation, stimulating adhesion and migration. These interactions involve cell surface receptor-ligand pairs on both cells. This list is likely incomplete as new interactions of importance to platelet biology are continuing to be discovered as illustrated by our discovery of PEAR-1 binding to FcεR1α. Results: We have developed a highly sensitive avidity-based assay to identify novel extracellular interactions among 126 recombinantly-expressed platelet cell surface and secreted proteins involved in platelet aggregation. In this study, we will use this method to identify novel platelet-monocyte interactions. We aim to identify ligands for orphan receptors and novel partners of well-known proteins. Identified interactions will be studied in preliminary functional assays to demonstrate relevance to the inflammatory processes supporting atherogenesis. Conclusions: Platelet-monocyte interactions are essential for the development of thromboinflammatory disease. Up until relatively recently, technologies only allow us to limit our studies on each individual protein interaction at a single time. These studies propose for the first time to study the cell surface platelet-monocyte interactions in a systematic large-scale approach using a reliable screening method we have developed. If successful, this will likely to identify previously unknown ligands for important receptors that will be investigated in details and also provide a list of novel interactions for the field. This should stimulate studies on developing alternative therapeutic strategies to treat vascular inflammatory disorders such as atherosclerosis, DVT and sepsis and other clinically important inflammatory conditions.

Keywords: membrane proteins, large-scale screening, platelets, recombinant expression

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623 Isolation and Molecular Detection of Marek’s Disease Virus from Outbreak Cases in Chicken in South Western Ethiopia

Authors: Abdela Bulbula

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Background: Marek’s disease virus is a devastating infection, causing high morbidity and mortality in chickens in Ethiopia. Methods: The current study was conducted from March to November, 2021 with the general objective of performing antemortem and postmortem, isolation, and molecular detection of Marek’s disease virus from outbreak cases in southwestern Ethiopia. Accordingly, based on outbreak information reported from the study sites namely, Bedelle, Yayo, and Bonga towns in southwestern Ethiopia, 50 sick chickens were sampled. The backyard and intensive farming systems of chickens were included in the sampling and priorities were given for chickens that showed clinical signs that are characteristics of Marek’s disease. Results: By clinical examinations, paralysis of legs and wings, gray eye, loss of weight, difficulty in breathing, and depression were recorded on all chickens sampled for this study and death of diseased chickens was observed. In addition, enlargement of the spleen and gross lesions of the liver and heart were recorded during postmortem examination. The death of infected chickens was observed in both vaccinated and non-vaccinated flocks. Out of 50 pooled feather follicle samples, Marek’s disease virus was isolated from 14/50 (28%) by cell culture method and out of six tissue samples, the virus was isolated from 5/6(83.30%). By Real time polymerization chain reaction technique, which was targeted to detect the Meq gene, Marek’s disease virus was detected from 18/50 feather follicles which accounts for 36% of sampled chickens. Conclusion: In general, the current study showed that the circulating Marek’s disease virus in southwestern Ethiopia was caused by the oncogenic Gallid herpesvirus-2 (Serotype-1). Further research on molecular characterization of revolving virus in current and other regions is recommended for effective control of the disease through vaccination.

Keywords: Ethioi, Marek's disease, isolation, molecular

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622 Fiduciary in Theory and Practice: The Perspective of the Allodial Rights Holders of Customary Land in Ghana

Authors: Kwasi Sarfo, Bernard Okoampah Otu

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The 1992 constitution of the Republic of Ghana recognises the authority and roles of traditional leaders and considers them as being entrusted with fiduciary responsibility over land in their respective territories. The new land act, Act 1036 of 2020, in buttressing the fiduciary role of traditional leaders in land matters, inserted the traditional leaders’ accountability clause. Many traditional leaders have expressed their misgivings about the insertion of this clause. Therefore, there appears to be a misunderstanding of the concept of fiduciary in land management in Ghana. The objective of this study is to examine the concept of fiduciary in respect of allodial rights holders in land management and administration and how this concept is perceived and applied by traditional leaders. The study seeks to provide insights into the discrepancy between fiduciary theory and its practical implementation in Ghana. The study is based on a qualitative empirical research approach and adopts in-depth interviews as a primary method of data collection. The study also adopts the theory of New Institutionalism of social anthropology in analysing and interpreting the findings. The data for this study was collected over a period of one year, from July 2022 to July 2023, as part of one of the author's PhD dissertation. The collected data were carefully analysed using the principles of thematic analysis, identifying key themes and patterns. This study does not seek to pursue the discussions from a legal standpoint but from a social anthropological perspective and argues that the concept of fiduciary in theory is far different from what pertains in practice and that traditional leaders’ assumptions and application of this concept contribute to the alienation of customary and communal land at the expense of their subjects. This study deepens our understanding of the continuous expropriation of communal landholders in many rural communities in Africa in the era of global land grabbing, which worsens their poverty levels. It also explains further the theory of new institutionalism of social anthropology by highlighting how the theory manifests in practice in the case of Ghana.

Keywords: fiduciary, customary land tenure, allodial rights, land alienation, communal land, Ghana

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621 Engineering of Reagentless Fluorescence Biosensors Based on Single-Chain Antibody Fragments

Authors: Christian Fercher, Jiaul Islam, Simon R. Corrie

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Fluorescence-based immunodiagnostics are an emerging field in biosensor development and exhibit several advantages over traditional detection methods. While various affinity biosensors have been developed to generate a fluorescence signal upon sensing varying concentrations of analytes, reagentless, reversible, and continuous monitoring of complex biological samples remains challenging. Here, we aimed to genetically engineer biosensors based on single-chain antibody fragments (scFv) that are site-specifically labeled with environmentally sensitive fluorescent unnatural amino acids (UAA). A rational design approach resulted in quantifiable analyte-dependent changes in peak fluorescence emission wavelength and enabled antigen detection in vitro. Incorporation of a polarity indicator within the topological neighborhood of the antigen-binding interface generated a titratable wavelength blueshift with nanomolar detection limits. In order to ensure continuous analyte monitoring, scFv candidates with fast binding and dissociation kinetics were selected from a genetic library employing a high-throughput phage display and affinity screening approach. Initial rankings were further refined towards rapid dissociation kinetics using bio-layer interferometry (BLI) and surface plasmon resonance (SPR). The most promising candidates were expressed, purified to homogeneity, and tested for their potential to detect biomarkers in a continuous microfluidic-based assay. Variations of dissociation kinetics within an order of magnitude were achieved without compromising the specificity of the antibody fragments. This approach is generally applicable to numerous antibody/antigen combinations and currently awaits integration in a wide range of assay platforms for one-step protein quantification.

Keywords: antibody engineering, biosensor, phage display, unnatural amino acids

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620 Influence of IL-1β on Hamster Blastocyst Hatching via Regulation of Hatching Associated Proteases

Authors: Madhulika Pathak, Polani Seshagiri, Vani Venkatappa

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Blastocyst hatching is an indispensable process for successful implantation. One of the major reasons for implantation failure in IVF clinic is poor quality of embryo, which are not development/hatching-competent. Therefore, attempts are required to develop or enhance the culture system with a molecule recapitulating the autocrine/paracrine factors containing the environment of in-vivo endometrial milieu. We have tried to explore the functional molecules involved in the hamster hatching phenomenon. Blastocyst hatching is governed by several molecules that are entwined and regulate this process, among which cytokines are known to be expressed and are still least explored. Two of such cytokines we have used for our study are IL-1β and its natural antagonist IL-1ra to understand the functional dynamics of cytokines involved in the hatching process. Using hamster, an intriguing experimental model for hatching behavior, we have shown the mRNA (qPCR) and protein (ICC) expression of IL-1β, IL-1ra and IL-1 receptor type 1 throughout all the stages of morula, blastocyst and hatched blastocyst. Post-asserting the expression, the functional role is shown by supplementation studies, where IL-1β supplementation showed enhancement in hatching level (IL-1β treated: 84.1 ± 4.2% vs control: 63.7 ± 3.1 %, N=11), further confirmed by the diminishing effect of IL-1ra on hatching rate (IL-1ra treated: 27.5 ± 11.1% vs control: 67.9 ± 3.1%). The exogenous supplementation of IL-1ra decreased the survival rate of embryos and affected the viability in dose-dependent manner, establishing the importance of IL-1β in blastocyst cell survival. Previously, the cathepsin L and B were established as the proteases that were involved in the hamster hatching process. The inducing effect of IL-1β was correlated with enhanced mRNA level, as analyzed by qPCR, for both CAT L (by 1.9 ± 0.5 fold) and CAT B (by 3.5 ± 0.1) fold which was diminished in presence of IL-1ra (Cat L by 88 percent and Cat B by 94 percent. Moreover, using a specific fluorescent substrate-based assay kit, the enzymatic activity of these proteases was found to be increased in presence of IL-1β (Cat L by 2.1 ± 0.1 fold and CAT B by 2.3 ± 0.7 fold) and was curtailed in presence of IL-1ra. These observations provide functional insights with respect to the involvement of cytokines in the hatching process. This has implications in understanding the hatching biology and improving the embryo development quality in IVF clinics.

Keywords: Blastocyst, Cytokines, Hatching, Interleukin

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619 Effects of Anti-FGL2 Monoclonal Antibody SPF89 on Vascular Inflammation

Authors: Ying Sun, Biao Cheng, Qing Lu, Xuefei Tao, Xiaoyu Lai, Cheng Guo, Dan Wang

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Fibrinogen-like protein 2 (FGL2) has recently been identified to play an important role in inflammatory diseases such as atherosclerosis through a thrombin-dependent manner. Here, a murine monoclonal antibody was raised against the critical residue Ser(89) of FGL2, and the effects of the anti-FGL2 mAb (SPF89) were analyzed in human umbilical vein endothelial cells (HUVECs) and THP-1 cells. Firstly, it was proved that SPF89, which belongs to the IgG1 subtype with a KD value of 44.5 pM, could specifically show the expression levels of protein FGL2 in different cell lines of known target gene status. The lipopolysaccharide (LPS)-mediated endothelial cell proliferation was significantly inhibited with a decline of phosphorylation nuclear factor-κB (NF-κB) in a dose-dependent manner after SPF89 treatment. Furthermore, SPF89 reduced LPS-induced expression of adhesion molecules and inflammatory cytokines such as vascular cell adhesion molecule-1, tumor necrosis factor-α, Matrix metalloproteinase MMP-2, Integrin αvβ3, and interleukin-6 in HUVECs. In macrophage-like THP-1 cells, SPF89 effectively inhibited LPS and low-density lipoprotein-induced foam cell formation. However, these anti-inflammatory and anti-atherosclerotic effects of anti-FGL2 mAb in HUVECs and THP-1 cells were significantly reduced after treatment with an NF-κB inhibitor PDTC. All the above suggest, by efficiently inhibiting LPS-induced pro-inflammatory effects in vascular endothelial cells by attenuating NF-κB dependent pathway, the new anti-FGL2 mAb SPF89 could to be a potential therapeutic candidate for protecting the vascular endothelium against inflammatory diseases such as atherosclerosis. This work was supported by the Program of Sichuan Science and Technology Department (2017FZ0069) and Collaborative Innovation Program of Sichuan for Elderly Care and Health(YLZBZ1511).

Keywords: monoclonal antibody, fibrinogen like protein 2, inflammation, endothelial cells

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618 Antibacterial Activities of Lactic Acid Bacteria on Potential Multidrug - Resistant Pathogens Isolated from Rabbit

Authors: Checkfaith I. Aizebeoje, Temitope O. Lawal, Bolanle A. Adeniyi

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The overuse and abuse of antibiotics in treating zoonotic infections in humans and opportunistic infections in rabbit has contributed to the increase in antimicrobial drug resistance, therefore, an alternative to antibiotics is needed in treating these infections. The study was carried out to determine the antimicrobial activity of lactic acid bacteria (LAB) isolated from rabbit’s faeces against multidrug-resistant (MDR) pathogens isolated from the same rabbit. Twelve faecal samples and twelve swabs from fur samples were randomly collected aseptically from apparently healthy rabbits from Ajibode, Ibadan and University of Ibadan research farm in Ibadan, Oyo state, Nigeria. Lactic acid bacteria and multidrug-resistant pathogens were isolated using appropriate agar media and identified by partial sequencing of the 16SrRNA gene. Antibiotic susceptibility pattern of isolated bacteria and LAB were determined by the agar diffusion method. The antibacterial activity of the LAB against the test pathogens was determined using the agar overlay and agar diffusion methods. The pathogens Myroides gitamensis, Citrobacter rodentium, Acinetobacter johnsonii, Enterobacter oryzendophyticus and Serratia marcescens as well as twenty-eight (28) species of LAB belonging to Acetobacter and Lactobacillus genera were identified and characterized. Lactobacillus plantarum had the highest (60.71%) occurrence of the LAB. Viable cells and cell free supernatant (CFS) of isolated LAB inhibited the growth of the test organisms with the largest zone of inhibition (40 mm) produced by Lactobacillus plantarum against Citrobacter rodentium. This study showed that LAB from rabbit possess considerable antibacterial activity against multidrug-resistant bacteria from the same environment.

Keywords: antibacterial activities, cell-free supernatant, lactic acid bacteria; multidrug-resistant pathogens, rabbits’ faeces

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617 Phylogeographic Reconstruction of the Tiger Shrimp (Penaeus monodon) Invasion in the Atlantic Ocean: The Role of the Farming Systems in the Marine Biological Invasions

Authors: Juan Carlos Aguirre Pabon, Stephen Sabatino, James Morris, Khor Waiho, Antonio Murias

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The tiger shrimp Penaeus monodon is one of the most important species in aquaculture and is native to the Indo-Pacific Ocean. During its greatest success in world production (70s and 80s) was introduced in many Atlantic Ocean countries for cultivation purposes and is currently reported as established in several countries of this area. Because there are no studies to understand the magnitude of the invasion process, this is an exciting opportunity to test evolutionary hypotheses in the context of marine invasions mediated by culture systems; therefore, the purpose of this study was to reconstruct the scenario of invasion of P. monodon in the Atlantic Ocean, by using mitochondrial DNA and eight loci microsatellites. In addition, samples of the invasion area in the Atlantic Ocean (US, Colombia, Venezuela, Brazil, Guienne Bissau, Senegal), the Indo-Pacific Ocean (Indonesia, India, Mozambique), and some cultivation systems (India, Bangladesh, Madagascar) were collected; and analysis of phylogenetic relationships (using some species of the family), genetic diversity, structure population, and demographic changes were performed. High intraspecific divergence in P. semisulcatus and P. monodon were found, high genetic variability in all sites (especially with microsatellites) and the presence of three clusters or populations. In addition, signs of demographic expansion in the culture population and bottlenecks in the invasive and native populations were found, as well as evidence of gene mixtures from all of the populations studied, implying that cropping systems play an essential role in mitigating the negative effects of the founder effect and providing a source of genetic variability that can ensure the success of the invasion.

Keywords: species introduction, increased variability, demographic changes, promoting invasion.

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616 Phenolic Composition and Antioxidant Activity of Sorbus L. Fruits and Leaves

Authors: Raudone Lina, Raudonis Raimondas, Gaivelyte Kristina, Pukalskas Audrius, Janulis Valdimaras, Viskelis Pranas

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Sorbus L. species are widely distributed in the Northern hemisphere and have been used for medicinal purposes in various traditional medicine systems and as food ingredients. Various Sorbus L. raw materials, fruits, leaves, inflorescences, barks, possess diuretic, anti-inflammatory, hypoglycemic, anti-diarrheal and vasoprotective activities. Phenolics, to whom main pharmacological activities are attributed, are compounds of interest due to their notable antioxidant activity. The aim of this study was to determine the antioxidant profiles of fruits and leaves of selected Sorbus L. species (S. anglica, S. aria f. latifolia, S. arranensis, S. aucuparia, S. austriaca, S. caucasica, S. commixta, S. discolor, S. gracilis, S. hostii, S. semi-incisa, S. tianschanica) and to identify the phenolic compounds with potent contribution to antioxidant activity. Twenty two constituents were identified in Sorbus L. species using ultra high performance liquid chromatography coupled to quadruple and time-of-flight mass spectrometers (UPLC–QTOF–MS). Reducing activity of individual constituents was determined using high performance liquid chromatography (HPLC) coupled to post-column FRAP assay. Signicantly greatest trolox equivalent values corresponding up to 45% of contribution to antioxidant activity were assessed for neochlorogenic and chlorogenic acids, which were determined as markers of antioxidant activity in samples of leaves and fruits. Characteristic patterns of antioxidant profiles obtained using HPLC post-column FRAP assay significantly depend on specific Sorbus L. species and raw materials and are suitable for equivalency research of Sorbus L. fruits and leaves. Selecting species and target plant organs with richest phenolic composition and strongly expressed antioxidant power is the first step in further research of standardized extracts.

Keywords: FRAP, antioxidant, phenolic, Sorbus L., chlorogenic acid, neochlorogenic acid

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615 pH and Temperature Triggered Release of Doxorubicin from Hydogen Bonded Multilayer Films of Polyoxazolines

Authors: Meltem Haktaniyan, Eda Cagli, Irem Erel Goktepe

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Polymers that change their properties in response to different stimuli (e.g. light, temperature, pH, ionic strength or magnetic field) are called ‘smart’ or ‘stimuli-responsive polymers’. These polymers have been widely used in biomedical applications such as sensors, gene delivery, drug delivery or tissue engineering. Temperature-responsive polymers have been studied extensively for controlled drug delivery applications. As regard of pseudo-peptides, poly (2-alky-2-oxazoline)s are considered as good candidates for delivery systems due to their stealth behavior and nontoxicity. In order to build responsive multilayer films for controlled drug release applications from surface, Layer by layer technique (LBL) is a powerful technique with an advantage of nanometer scale control over spatial architecture and morphology. Multilayers can be constructed on surface where non-covalent interactions including electrostatic interactions, hydrogen bonding, and charge-transfer or hydrophobic-hydrophobic interactions. In the present study, hydrogen bounded multilayer films of poly (2-alky-2-oxazoline) s with tannic acid were prepared in order to use as a platform to release Doxorubicin (DOX) from surface with pH and thermal triggers. For this purpose, poly (2-isopropyl-2-oxazoline) (PIPOX) and poly (2-ethyl-2-oxazoline) (PETOX) were synthesized via cationic ring opening polymerization (CROP) with hydroxyl end groups. Two polymeric multilayer systems ((PETOX)/(DOX)-(TA) complexes and (PIPOX)/(DOX)-(TA) complexes) were designed to investigate of controlled release of Doxorubicin (DOX) from surface with pH and thermal triggers. The drug release profiles from the multilayer thin films with alterations of pH and temperature will been examined with UV-Vis Spectroscopy and Fluorescence Spectroscopy.

Keywords: temperature responsive polymers, h-bonded multilayer films, drug release, polyoxazoline

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614 AMF activates PDH 45 and G-proteins Genes to Alleviate Abiotic Stress in Tomato Plants

Authors: Deepak Bhardwaj, Narendra Tuteja

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Global climate change is impacting large agrarian societies, especially those in countries located near the equator. Agriculture, and consequently, plant-based food, is the hardest hit in tropical and sub-tropical countries such as India due to an increased incidence of drought as well as an increase in soil salinity. One method that holds promise is AMF-rich biofertilizers which assist in activating proteins which in turn help alleviate abiotic stress in plants. In the present study, we identified two important species of (arbuscular mycorrhizal fungus) AMF belonging to Glomus and Gigaspora from the rhizosphere of the important medicinal plant Justicia adathoda. These two species have been found to be responsible for the abundance of Justicia adathoda in the semi-arid areas of the Jammu valley located in northern India, namely, the Union Territory of Jammu and Kashmir. We isolated the species of Glomus and Gigaspora from the rhizosphere of Justicia adathoda and used them as biofertilizers for the tomato plant. Significant improvements in the growth parameters were observed in the tomato plants inoculated with Glomus sp. and Gigaspora sp. in comparison with the tomato plants that were grown without AMF treatments. Tomato plants grown along with Glomus sp. and Gigaspora sp. have been observed to withstand 200 mM of salinity and 25% PEG stress. AMF also resulted in an increased concentration of proline and antioxidant enzymes in tomato plants. We also examined the expression levels of salinity and drought stress-inducible genes such as pea DNA helicase 45 (PDH 45) and genes of G-protein subunits of the tomato plants inoculated with and without AMF under stress and normal conditions. All the stress-inducible genes showed a significant increase in their gene expression under stress and AMF inoculation, while their levels were found to be normal under AMF inoculation without stress. We propose a model of abiotic stress alleviation in tomato plants with the help of external factors such as AMF and internally with the help of proteins like PDH 45 and G-proteins.

Keywords: AMF, abiotic stress, g-proteins, PDH-45

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613 A Rare Entity: Case Report on Anaesthetic Management in Robinow Syndrome

Authors: Vidhi Chandra, Arshpreet Singh Grewal

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A five-year-old male child born from non-consanguineous marriage, who presented with complaints of growth retardation and no appreciable increase in the penile size since birth and he was posted for de-gloving of penis with dissection of corpora under anaesthesia. After thorough preoperative evaluation it was revealed that patient had peculiar facial dysmorphism that of Robinow Syndrome, high arched palate, Mallampati grade III, mesomelic limbs, scoliotic spine and short stature. All routine investigation were within normal limit, electrocardiography (ECG) and 2D-Echocardiography (ECHO) were normal. In antero-posterior roentgenogram chest showed butterfly and hemivertebrae at multiple levels. The patient was considered to be ASA II. On the day of surgery after ensuring fasting of 6 hours, patient was taken in operation theatre, all standard ASA monitoring was done with ECG, non-invasive blood pressure, peripheral oxygen saturation (SpO2) and body temperature. The patient was pre-oxygenated with 100% oxygen with anatomical face mask. General anaesthesia was induced with Sevoflurane 1-8%, and airway was secured with an appropriate size supraglottic airway and anaesthesia was maintained with nitrous oxide and oxygen in 1:1 ratio along with sevoflurane 2%. An ultrasound guided caudal block was given owing to the skeletal deformities making it difficult even under USG guidance. Post operatively patient was given supportive care with proper hydration, antibiotics, anti-inflammatory and analgesics. He was discharged the next day and followed up weekly for a month. DISCUSSION Robinow syndrome is genetically inherited as autosomal dominant, autosomal recessive or heterogenous disorder involving tyrosine kinase ROR2 gene located on chromosome 9. It has low incidence with no preponderance for any gender. Though intelligence is normal but developmental delay and mental retardation occurs in 20%cases

Keywords: Robinow Syndrome, dwarfism, paediatric, anaesthesia

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612 The Impact of the Fitness Center Ownership Structure on the Service Quality Perception in the Fitness in Serbia

Authors: Dragan Zivotic, Mirjana Ilic, Aleksandra Perovic, Predrag Gavrilovic

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As with the provision of other services, the service quality perception is one of the key factors that the modern manager must pay attention to. Countries in which the state regulation is in transition also have specific features in providing fitness services. Identification of the dimensions in which the most significant different service quality perception between different types of fitness centers, enables managers to profile the offer according to the wishes and expectations of users. The aim of the paper was the comparison of the quality of services perception in the field of fitness in Serbia between three categories of fitness centers: the privately owned centers, the publicly owned centers, and the Public-private partnership centers. For this research 350 respondents of both genders (174 men and 176 women) were interviewed, aged between 18 and 68 years, being beneficiaries of fitness services for at least 1 year. Administered questionnaire with 100 items provided information about the 15 basic areas in which they expressed the service quality perception in the gym. The core sample was composed of 212 service users in private fitness centers, 69 service users in public fitness centers and 69 service users in the public-private partnership. Sub-samples were equal in representation of women and men, as well as by age and length of use of fitness services. The obtained results were subject of univariate analysis with the Kruskal-Wallis non-parametric analysis of variance. Significant differences between the analyzed sub-samples were not found solely in the areas of rapid response and quality outcomes. In the multivariate model, the results were processed by backward stepwise discriminant analysis that extracted 3 areas that maximize the differences between sub-samples: material and technical basis, secondary facilities and coaches. By applying the classification function 93.87% of private centers services users, 62.32% of public centers services users and 85.51% of the public-private partnership centers users of services were correctly classified (total 86.00%). These results allow optimizing the allocation of the necessary resources in profiling offers of a fitness center in order to optimally adjust it to the user’s needs and expectations.

Keywords: fitness, quality perception, management, public ownership, private ownership, public-private partnership, discriminative analysis

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611 Structural and Functional Characterization of the Transcriptional Regulator Rv1176 of Mycobacterium tuberculosis H37Rv

Authors: Vikash Yadav, Ashish Arora

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Microorganisms have self-defense mechanisms to protect themselves from toxic environments. Phenolic acid decarboxylase(pad) is responsible for the defense against toxicity caused by phenolic acids, converting them into less toxic vinyl derivatives. The transcription of the pad gene is regulated by a negative transcription factor, phenolic acid decarboxylase regulators (PadR), in a substrate-inducible manner. The PadR family members share the conserved DNA-binding features and interact with the operator DNA using a winged helix-turn-helix (wHTH) motif, which contains a three-helix motif and a β-stranded wing. The members of this family function as transcriptional regulators that are involved in various cellular survival processes, such as toxin production, detoxification, multidrug resistance, antibiotic biosynthesis, and carbon catabolism. Rv1176 of Mycobacterium tuberculosis H37Rv has been assigned to the PadR family protein that remains to be structurally and functionally uncharacterized. To reveal the structural mechanism by which Rv1176 could regulates effector-responsive transcription, several experiments were performed, including Electrophoretic Mobility Shift Assay (EMSA) for DNA protein interaction, differential scanning calorimetry (DSC) and Differential Scanning Fluorimetry (DSF) for temperature and ligand-dependent protein stability, Circular Dichroism (CD) spectroscopy for secondary structure analysis. Further, to evaluate the functional role of Rv1176, the intracellular survival of recombinant M. smegmatis was examined in murine macrophage cell line J774A.1 and different stressed conditions like oxidative, pH, and nutritive stress. All these studies demonstrated that Rv1176 could behave as a transcription regulator and its expression in recombinant M. smegmatis increases intracellular survival.

Keywords: EMSA, Mycobacterium tuberculosis, PadR family protein, transcriptional regulator

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610 LTF Expression Profiling Which is Essential for Cancer Cell Proliferation and Metastasis, Correlating with Clinical Features, as Well as Early Stages of Breast Cancer

Authors: Azar Heidarizadi, Mahdieh Salimi, Hossein Mozdarani

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Introduction: As a complex disease, breast cancer results from several genetic and epigenetic changes. Lactoferrin, a member of the transferrin family, is reported to have a number of biological functions, including DNA synthesis, immune responses, iron transport, etc., any of which could play a role in tumor progression. The aim of this study was to investigate the bioinformatics data and experimental assay to find the pattern of promoter methylation and gene expression of LTF in breast cancer in order to study its potential role in cancer management. Material and Methods: In order to evaluate the methylation status of the LTF promoter, we studied the MS-PCR and Real-Time PCR on samples from patients with breast cancer and normal cases. 67 patient samples were conducted for this study, including tumoral, plasma, and normal tissue adjacent samples, as well as 30 plasma from normal cases and 10 tissue breast reduction cases. Subsequently, bioinformatics analyses such as cBioPortal databases, string, and genomatix were conducted to disclose the prognostic value of LTF in breast cancer progression. Results: The analysis of LTF expression showed an inverse relationship between the expression level of LTF and the stages of tissues of breast cancer patients (p<0.01). In fact, stages 1 and 2 had a high expression in LTF, while, in stages 3 and 4, a significant reduction was observable (p < 0.0001). LTF expression frequently alters with a decrease in the expression in ER⁺, PR⁺, and HER2⁺ patients (P < 0.01) and an increase in the expression in the TNBC, LN¯, ER¯, and PR- patients (P < 0.001). Also, LTF expression is significantly associated with metastasis and lymph node involvement factors (P < 0.0001). The sensitivity and specificity of LTF were detected, respectively. A negative correlation was detected between the results of level expression and methylation of the LTF promoter. Conclusions: The altered expression of LTF observed in breast cancer patients could be considered as a promotion in cell proliferation and metastasis even in the early stages of cancer.

Keywords: LTF, expression, methylation, breast cancer

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609 Whole Exome Sequencing Data Analysis of Rare Diseases: Non-Coding Variants and Copy Number Variations

Authors: S. Fahiminiya, J. Nadaf, F. Rauch, L. Jerome-Majewska, J. Majewski

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Background: Sequencing of protein coding regions of human genome (Whole Exome Sequencing; WES), has demonstrated a great success in the identification of causal mutations for several rare genetic disorders in human. Generally, most of WES studies have focused on rare variants in coding exons and splicing-sites where missense substitutions lead to the alternation of protein product. Although focusing on this category of variants has revealed the mystery behind many inherited genetic diseases in recent years, a subset of them remained still inconclusive. Here, we present the result of our WES studies where analyzing only rare variants in coding regions was not conclusive but further investigation revealed the involvement of non-coding variants and copy number variations (CNV) in etiology of the diseases. Methods: Whole exome sequencing was performed using our standard protocols at Genome Quebec Innovation Center, Montreal, Canada. All bioinformatics analyses were done using in-house WES pipeline. Results: To date, we successfully identified several disease causing mutations within gene coding regions (e.g. SCARF2: Van den Ende-Gupta syndrome and SNAP29: 22q11.2 deletion syndrome) by using WES. In addition, we showed that variants in non-coding regions and CNV have also important value and should not be ignored and/or filtered out along the way of bioinformatics analysis on WES data. For instance, in patients with osteogenesis imperfecta type V and in patients with glucocorticoid deficiency, we identified variants in 5'UTR, resulting in the production of longer or truncating non-functional proteins. Furthermore, CNVs were identified as the main cause of the diseases in patients with metaphyseal dysplasia with maxillary hypoplasia and brachydactyly and in patients with osteogenesis imperfecta type VII. Conclusions: Our study highlights the importance of considering non-coding variants and CNVs during interpretation of WES data, as they can be the only cause of disease under investigation.

Keywords: whole exome sequencing data, non-coding variants, copy number variations, rare diseases

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608 Support for Privilege Based on Nationality in Switched-At-Birth Scenario

Authors: Anne Lehner, Mostafa Salari Rad, Jeremy Ginges

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Many of life’s privileges (and burdens) are thrust on us at birth. Someone born white or male in the United States is also born with a set of advantages over someone born non-white or female. One aspect of privileges conferred by birth is that they are so entrenched in social institutions and social norms that until they are robustly challenged, they can be seen as a moral good. While American society increasingly confronts privileges based on gender and race, other types of privileges, like one's nationality, see less attention. The nationality one is born into can have enormous effects on one’s personal life, work opportunities, and health outcomes. Yet, we predicted that although most Americans would regard it as absurd to think that white people have a right to protect their privileges and 'way of life', they would regard it as obvious that Americans have a right to protect the American way of life and associated privileges. In a preregistered study we presented 300 Americans randomly with one out of three 'privilege scales' in order to assess their agreement with certain statements. The domains for the privilege scales were nationality, race, and gender. Next, all participants completed the switched-at-birth task assessing ones tendency to essentialize nationality. We found that Americans are more approving of privilege based on nationality than of privilege based on gender and race. In addition, we found an interaction of condition with ideology, showing that conservatives are in general more approving of the privilege of any kind than liberals are, and they especially approve of privilege based on nationality. For the switched-at-birth task, we found that both, liberals as well as conservatives are equally willing to grant the child 100% American nationality. Whether or not one chose 100% is unrelated to the expressed approval of privilege based on nationality. One might hesitate to fully grant the child 100% American nationality in the task, yet disapprove of privilege based on nationality. This shows that as much as we see beholders of privilege being oblivious to their statuses within other social categories, like gender or race, we seem to detect the same blindness for the privilege based on nationality. Liberals showing relatively fewer support for privilege based on nationality compared to conservatives still refused to acknowledge the child as having become 100% American and thereby denying the privileges it potentially bestows upon them.

Keywords: thought experiment, anti-immigrant attitudes, privilege of nationality, immigration, moral circles, psychology

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607 Detection of Cryptosporidium Oocysts by Acid-Fast Staining Method and PCR in Surface Water from Tehran, Iran

Authors: Mohamad Mohsen Homayouni, Niloofar Taghipour, Ahmad Reza Memar, Niloofar Khalaji, Hamed Kiani, Seyyed Javad Seyyed Tabaei

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Background and Objective: Cryptosporidium is a coccidian protozoan parasite; its oocysts in surface water are a global health problem. Due to the low number of parasites in the water resources and the lack of laboratory culture, rapid and sensitive method for detection of the organism in the water resources is necessarily required. We applied modified acid-fast staining and PCR for the detection of the Cryptosporidium spp. and analysed the genotypes in 55 samples collected from surface water. Methods: Over a period of nine months, 55 surface water samples were collected from the five rivers in Tehran, Iran. The samples were filtered by using cellulose acetate membrane filters. By acid fast method, initial identification of Cryptosporidium oocyst were carried out on surface water samples. Then, nested PCR assay was designed for the specific amplification and analysed the genotypes. Results: Modified Ziehl-Neelsen method revealed 5–20 Cryptosporidium oocysts detected per 10 Liter. Five out of the 55 (9.09%) surface water samples were found positive for Cryptosporidium spp. by Ziehl-Neelsen test and seven (12.7%) were found positive by nested PCR. The staining results were consistent with PCR. Seven Cryptosporidium PCR products were successfully sequenced and five gp60 subtypes were detected. Our finding of gp60 gene revealed that all of the positive isolates were Cryptosporidium parvum and belonged to subtype families IIa and IId. Conclusion: Our investigations were showed that collection of water samples were contaminated by Cryptosporidium, with potential hazards for the significant health problem. This study provides the first report on detection and genotyping of Cryptosporidium species from surface water samples in Iran, and its result confirmed the low clinical incidence of this parasite on the community.

Keywords: Cryptosporidium spp., membrane filtration, subtype, surface water, Iran

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606 Predictive Value of Hepatitis B Core-Related Antigen (HBcrAg) during Natural History of Hepatitis B Virus Infection

Authors: Yanhua Zhao, Yu Gou, Shu Feng, Dongdong Li, Chuanmin Tao

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The natural history of HBV infection could experience immune tolerant (IT), immune clearance (IC), HBeAg-negative inactive/quienscent carrier (ENQ), and HBeAg-negative hepatitis (ENH). As current biomarkers for discriminating these four phases have some weaknesses, additional serological indicators are needed. Hepatits B core-related antigen (HBcrAg) encoded with precore/core gene contains denatured HBeAg, HBV core antigen (HBcAg) and a 22KDa precore protein (p22cr), which was demonstrated to have a close association with natural history of hepatitis B infection, but no specific cutoff values and diagnostic parameters to evaluate the diagnostic efficacy. This study aimed to clarify the distribution of HBcrAg levels and evaluate its diagnostic performance during the natural history of infection from a Western Chinese perspective. 294 samples collected from treatment-naïve chronic hepatitis B (CHB) patients in different phases (IT=64; IC=72; ENQ=100, and ENH=58). We detected the HBcrAg values and analyzed the relationship between HBcrAg and HBV DNA. HBsAg and other clinical parameters were quantitatively tested. HBcrAg levels of four phases were 9.30 log U/mL, 8.80 log U/mL, 3.00 log U/mL, and 5.10 logU/mL, respectively (p < 0.0001). Receiver operating characteristic curve analysis demonstrated that the area under curves (AUCs) of HBcrAg and quantitative HBsAg at cutoff values of 9.25 log U/mL and 4.355 log IU/mL for distinguishing IT from IC phases were 0.704 and 0.694, with sensitivity 76.39% and 59.72%, specificity 53.13% and 79.69%, respectively. AUCs of HBcrAg and quantitative HBsAg at cutoff values of 4.15 log U/mlmL and 2.395 log IU/mlmL for discriminating between ENQ and ENH phases were 0.931 and 0.653, with sensitivity 87.93% and 84%, specificity 91.38% and 39%, respectively. Therefore, HBcrAg levels varied significantly among four natural phases of HBV infection. It had higher predictive performance than quantitative HBsAg for distinguishing between ENQ-patients and ENH-patients and similar performance with HBsAg for the discrimination between IT and IC phases, which indicated that HBcrAg could be a potential serological marker for CHB.

Keywords: chronic hepatitis B, hepatitis B core-related antigen, hepatitis B surface antigens, hepatitis B virus

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605 Rohingya Resettlement Roadblocks: Challenges and Potentials

Authors: Ishrat Zakia Sultana

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The solution to the Rohingya crisis has become complicated than it was anticipated. Because of consistent persecution, ethnic cleansing, and genocide against the Rohingya in Burma, four major influxes of the Rohingya people took place to the neighboring country Bangladesh. After the latest influx of October 2016 and August 2017, the total number of Rohingya in Bangladesh stands somewhere between 900,000 to over one million, placing Bangladesh much ahead with the number of refugees compared to Dadaab and Kakuma in Kenya, Bidibidi in Uganda, and Zaatari in Jordan. While Bangladesh received recognition and appreciation for receiving such a large number of Rohingya, eventually finding a solution to the Rohingya crisis has become a serious problem. The host country and the Rohingya themselves long for repatriation, the most desired solution to the crisis. But going back to their own country is now almost an impossible matter due to the unwillingness of the Myanmar government. The other two options to the solution to Rohingya crisis – reintegration in the host country and third country resettlement – have drawn little attention until now. On the one hand, the geopolitical factors have been making the Rohingya crisis complex. On the other, the war and conflict between Russia-Ukraine and Palestine-Israel have lessening the importance of the Rohingya issue and been diverting the world’s attention from the Rohingya crisis. Clearly, without the support of international community, Bangladesh finds no sustainable way to repatriate 1.1 million Rohingya. Yet, possibilities of a third country resettlement remain unexplored. In the past few years, some countries have expressed interest in accepting the Rohingya as part of third country resettlement but the number they wanted to take is like a drop in the ocean. This paper examines the roadblocks for third country resettlement of the Rohingya. It aims to look at the underlying reasons for which international community is less interested in accepting the Rohingya as refugees. Is it the racial and religious identity of the Rohingya that are considered problematic to the resettlement process? In what ways geopolitical complexities affecting the resettlement issue? How do the Rohingya view third country resettlement? This paper looks for the answers to these questions. The paper is based on qualitative study conducted from 2016-2018 and 2021-2023 in Rohingya camps in Cox’s Bazar, Bangladesh. The camp management authority, the Rohingya themselves, and the NGOs working in the camp participated in the study.

Keywords: rohingya, refugee, resettlement, bangladesh

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604 Effects of Garlic and Stevia Extract Following Aerobic Exercise on Hypothalamic Semaphorin 4A and Plexin D1 Genes Expression in High-Fat Diet-Induced Obese Rats

Authors: Sayyed-Javad Ziaolhagh, Mojtaba Hokmabadi

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Introduction: Childhood obesity is a serious medical condition that affects children and adolescents even in the central nervous system. Semaphorins also play a role in the inflammatory process of the nervous system. On the other hand, it has been stated that garlic and stevia extracts following aerobic exercise are effective on immune system inflammation in addition to aerobic activity. Materials and Methods: For 15 weeks, 50 3-week-old male Wistar rats were fed with conventional rodent chow for control and a high-fat diet to induce obesity. Obese rats then were randomly assigned into 7 groups (n=5) based on the Lee index: healthy control (C), obese (OBS), obese + garlic (OBS+GAR), obese + Stevia (OBS+STV), obese + aerobic exercise (OBS+EXE), obese + garlic + aerobic exercise (OBS+GAR+EXE), and obese + stevia + aerobic exercise (OBS+STV+EXE). Training groups completed a progressive aerobic running program (at 8-15 m/min, 5-20 min/day, 5 days/week), and Stevia and garlic extract group (250 mg/kg/day, 5 days/week) were given orally once a day. Real-time PCR was used to determine the levels of Semaphorin 4A, and Plexin D1 gene expressions in the hypothalamus. Fold change analysis with ANOVA was performed for statistical analysis, with a significance threshold of P<0.05. Results: Body weight increased significantly in OBS compared to C (p= 0.013), but was not significantly changed in all treatment rats. Moreover, Semaphorin 4A was significantly increased in obese compared to control group (p= 0.041) and after 8 weeks, stevia extract (p=0.006), aerobic exercise (p=0.012) and garlic extract + aerobic exercise (p=0.008) significantly decreased compared to obese rats. In addition, Plexin D1 genes were also found in the hypothalamus of both obese and control rats but were insignificantly up-regulated when compared with the obese group (p=0.950). Conclusion: High-fat diet caused neuroinflammation by elevation of sema4A in obese rats and stevia, stevia with aerobic and garlic with aerobic could reduce this inflammation in rats. Also, none of them could alter Plexin D1.

Keywords: sema 4A, plexin D1, garlic, stevia

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603 Tissue-Specific Distribution of Cytochrome P450 1A1 and 3A in Rainbow Trout (Oncorhynchus mykiss)

Authors: Viktoriia Burkina, Vladimir Zlabek, Galia Zamaratskaia

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Cytochromes P450 (CYP) are important family of enzymes in Phase I metabolism. Environmental pollutants often act as inducers of the gene expression and activities CYP1A1 and CYP3A-like isoforms in fish. The activities are generally measured in the fish liver or gills, and less is known about tissue distribution of expression. In present study, the CYP1A1 and CYP3A-like activities were measured in rainbow trout liver, gill, intestine, heart, brain and gonads. The activities of CYP1A1 and CYP3A-like proteins were estimated as the rates of 7-ethoxyresorufin-O-deethylation (EROD) and benzyloxy-4-trifluoromethylcoumarin-O-debenzyloxylation (BFCOD), respectively. The CYP1A1 and CYP3A-like activities were detectable in all investigated fish tissues, with the highest activity in hepatic tissue followed by heart > brain > gill > intestine > gonads. To confirm the presence of CYP1A1 in different tissues, EROD activity was measured in presence of the selective inhibitors ellipticine (CYP1A1), ketoconazole (CYP3A), 8-methoxypsoralen (human CYP2A) and diallyl sulphide (CYP2E1). It was found that ellipticine, ketoconazole and 8-methoxypsoralen inhibited hepatic EROD activity by 88-98%. Ellipticine inhibited gill, intestine, and gonad EROD activity by 50%. In conclusion, EROD and BFCOD activities were detected in rainbow trout liver, gill, intestine, heart, brain and gonads. Further studies are needed to fully identify all CYP450 isoforms responsible for these activities. Acknowledgement: The study was financially supported by the Ministry of Education, Youth and Sports of the Czech Republic - projects „CENAKVA “(No. CZ.1.05/2.1.00/01.0024), “CENAKVA Center Development “(No. CZ.1.05/2.1.00/19.0380), “CENAKVA II “(No. LO1205 under the NPU I program), and "Development of USB - International mobility (No. CZ.02.2.69/0.0/0.0/16_027/0008364).

Keywords: BFCOD, EROD, fish, phase I metabolism, selective inhibitors

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602 Water Access and Food Security: A Cross-Sectional Study of SSA Countries in 2017

Authors: Davod Ahmadi, Narges Ebadi, Ethan Wang, Hugo Melgar-Quiñonez

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Compared to the other Least Developed Countries (LDCs), major countries in sub-Saharan Africa (SSA) have limited access to the clean water. People in this region, and more specifically females, suffer from acute water scarcity problems. They are compelled to spend too much of their time bringing water for domestic use like drinking and washing. Apart from domestic use, water through affecting agriculture and livestock contributes to the food security status of people in vulnerable regions like SSA. Livestock needs water to grow, and agriculture requires enormous quantities of water for irrigation. The main objective of this study is to explore the association between access to water and individuals’ food security status. Data from 2017 Gallup World Poll (GWP) for SSA were analyzed (n=35,000). The target population in GWP is the entire civilian, non-institutionalized, aged 15 and older population. All samples selection is probability based and nationally representative. The Gallup surveys an average of 1,000 samples of individuals per country. Three questions related to water (i.e., water quality, availability of water for crops and availability of water for livestock) were used as the exposure variables. Food Insecurity Experience Scale (FIES) was used as the outcome variable. FIES measures individuals’ food security status, and it is composed of eight questions with simple dichotomous responses (1=Yes and 0=No). Different statistical analyses such as descriptive, crosstabs and binary logistic regression, form the basis of this study. Results from descriptive analyses showed that more than 50% of the respondents had no access to enough water for crops and livestock. More than 85% of respondents were categorized as “food insecure”. Findings from cross-tabulation analyses showed that food security status was significantly associated with water quality (0.135; P=0.000), water for crops (0.106; P=0.000) and water for livestock (0.112; P=0.000). In regression analyses, the probability of being food insecure increased among people who expressed no satisfaction with water quality (OR=1.884 (OR=1.768-2.008)), not enough water for crops (OR=1.721 (1.616-1.834)) and not enough water for livestock (OR=1.706 (1.819)). In conclusion, it should note that water access affects food security status in SSA.

Keywords: water access, agriculture, livestock, FIES

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601 Cratoxy Formosum (Jack) Dyer Leaf Extract-Induced Human Breast and Liver Cancer Cells Death

Authors: Benjaporn Buranrat, Nootchanat Mairuae

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Cratoxylum formosum (Jack) Dyer (CF) has been used for the traditional medicines in South East Asian and Thailand. Normally, northeast Thai vegetables have proven cytotoxic to many cancer cells. Therefore, the present study aims to explore the molecular mechanisms underlying CF-induced cancer cell death and apoptosis on breast and liver cancer cells. The cytotoxicity and antiproliferative effects of CF on the human breast MCF-7 and liver HepG2 cancer cell lines were evaluated using sulforhodamine B assay and colony formation assay. Cell migration assay was measured using wound healing assay. The apoptosis induction mechanisms were investigated through reactive oxygen species formation, caspase 3 activity, and JC-1 activity. Gene expression by real-time PCR and apoptosis related protein levels by Western blot analysis. CF induced MCF-7 and HepG2 cell death by time- and dose-dependent manner. Furthermore, CF had the greater cytotoxic potency on MCF-7 more than HepG2 cells with IC50 values of 85.70+4.52 μM and 219.03±9.96 μM respectively, at 24 h. Treatment with CF also caused a dose-dependent decrease in colony forming ability and cell migration, especially on MCF-7 cells. CF induced ROS formation, increased caspase 3 activities, and decreased the mitochondrial membrane potential, and causing apoptotic body production and DNA fragmentation. CF significantly decreased expression of the cell cycle regulatory protein RAC1 and downstream proteins, cdk6. Additionally, CF enhanced p21 and reduced cyclin D1 protein levels. CF leaf extract induced cell death, apoptosis, antimigration in both of MCF-7 and HepG2 cells. CF could be useful for developing to anticancer drug candidate for breast and liver cancer therapy.

Keywords: cratoxylum formosum (jack) dyer, breast cancer, liver cancer, cell death

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600 Ectoine: A Compatible Solute in Radio-Halophilic Stenotrophomonas sp. WMA-LM19 Strain to Prevent Ultraviolet-Induced Protein Damage

Authors: Wasim Sajjad, Manzoor Ahmad, Sundas Qadir, Muhammad Rafiq, Fariha Hasan, Richard Tehan, Kerry L. McPhail, Aamer Ali Shah

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Aim: This study aims to investigate the possible radiation protective role of a compatible solute in the tolerance of radio-halophilic bacterium against stresses, like desiccation and exposure to ionizing radiation. Methods and Results: Nine different radio-resistant bacteria were isolated from desert soil, where strain WMA-LM19 was chosen for detailed studies on the basis of its high tolerance for ultraviolet radiation among all these isolates. 16S rRNA gene sequencing indicated that the bacterium was closely related to Stenotrophomonas sp. (KT008383). A bacterial milking strategy was applied for extraction of intracellular compatible solutes in 70% (v/v) ethanol, which were purified by high-performance liquid chromatography (HPLC). The compound was characterized as ectoine by 1H and 13C nuclear magnetic resonance (NMR), and mass spectrometry (MS). Ectoine demonstrated more efficient preventive activity (54.80%) to erythrocyte membranes and also inhibited oxidative damage to proteins and lipids in comparison to the standard ascorbic acid. Furthermore, a high level of ectoine-mediated protection of bovine serum albumin against ionizing radiation (1500-2000 Jm-2) was observed, as indicated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis. Conclusion: The results indicated that ectoine can be used as a potential mitigator and radio-protective agent to overcome radiation- and salinity-mediated oxidative damage in extreme environments. Significance and Impact of the Study: This study shows that ectoine from radio-halophiles can be used as a potential source in topical creams as sunscreen. The investigation of ectoine as UV protectant also changes the prospective that radiation resistance is specific only to molecular adaptation.

Keywords: ectoine, anti-oxidant, stenotrophomonas sp., ultraviolet radiation

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599 Embodied Spirituality in Gestalt Therapy

Authors: Silvia Alaimo

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This lecture brings to our attention the theme of spirituality within Gestalt therapy’s theoretical and clinical perspectives and which is closely connected to the fertile emptiness and creative indifference’ experiences. First of all, the premise that must be done is the overcoming traditional western culture’s philosophical and religious misunderstandings, such as the dicotomy between spirituality and pratical/material daily life, as well as the widespread secular perspective of classic psychology. Even fullness and emptiness have traditionally been associated with the concepts of being and not being. "There is only one way through which we can contact the deepest layers of our existence, rejuvenate our thinking and reach intuition (the harmony of thought and being): inner silence" (Perls) *. Therefore, "fertile void" doesn't mean empty in itself, but rather an useful condition of every creative and responsible act, making room for a deeper dimension close to spirituality. Spirituality concerns questions about the meaning of existence, which lays beyond the concrete and literal dimension, looking for the essence of things, and looking at the value of personal experience. Looking at fundamentals of Gestalt epistemology, phenomenology, aesthetics, and the relationship, we can reach the heart of a therapeutic work that takes spiritual contours and which are based on an embodied (incarnate size), through the relational aesthetic knowledge (Spagnuolo Lobb ), the deep contact with each other, the role of compassion and responsibility, as the patient's recognition criteria (Orange, 2013) rooted in the body. The aesthetic dimension, like the spiritual dimension to which it is often associated, is a subtle dimension: it is the dimension of the essence of things, of their "soul." In clinical practice, it implies that the relationship between therapist and patient is "in the absence of judgment," also called "zero point of creative indifference," expressed by ‘therapeutic mentality’. It consists in following with interest and authentic curiosity where the patient wants to go and support him in his intentionality of contact. It’s a condition of pure and simple awareness, of the full acceptance of "what is," a moment of detachment from one's own life in which one does not take oneself too seriously, a starting point for finding a center of balance and integration that brings to the creative act, to growth, and, as Perls would say, to the excitement and adventure of living.

Keywords: spirituality, bodily, embodied aesthetics, phenomenology, relationship

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598 An Evaluation of a Student Peer Mentoring Program

Authors: Nazeema Ahmed

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This paper reports on the development of a student peer mentoring programme at a higher education institution. The programme is dependent on volunteering senior undergraduate students who are trained to mentor first-year students studying towards an engineering degree. The evaluation of the programme took the form of first-year students completing a self-report paper questionnaire at the onset of a lecture and mentors completing their questionnaire electronically. The evaluation yielded mixed findings. Peer mentoring clearly benefited some students in their adjustment to the institution. Specific mentors’ personal attributes enabled the establishment of successful mentoring relationships, where encouragement, advice and academic assistance was provided. Gains were reciprocal with mentors reporting that the programme contributed towards their personal development. Confidence in the programme was expressed in mentors feeling that it was an initiative worth continuing and first-year students agreeing that it be recommended to future first-year students. This was despite many unfavourable experiences of mentors where their professionalism and commitment to the programme was suspect. It is evident that while mentors began with noble intentions they appear either to lose interest or become overwhelmed with their own workload as the academic year progresses. On the other hand, some mentors reported feeling challenged by the apathy of first-year students who failed to maximise the opportunity available to them. The different attitudes towards mentoring that manifested as a mentoring culture in some departments were particularly pertinent to its successful implementation. The findings point to the key role of academic staff in the mentoring programme who model the mentoring relationship in their interaction with student mentors. While their involvement in the programme may be perceived as a drain on resources in an already demanding academic teaching environment, it is imperative that structural changes be put in place for the programme to be both efficient and sustainable. A pervasive finding concerns the evolving institutional culture of student development in the faculty. Mentors and first-year students alike alluded to the potential of the mentoring programme provided it is seriously endorsed at both the departmental and faculty level. The findings provide a foundation from which to develop the programme further and to begin improving its capacity for maximizing student retention in South African higher education.

Keywords: engineering students, first-year students, peer mentoring

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597 Mannosidase Alpha Class 1B Member 1 Targets F Severe Acute Respiratory Syndrome Coronavirus 2 Spike Protein and Ebola Virus Glycoprotein to Endoplasmic Reticulum-To-Lysosome-Associated Degradation by Micro-Endoplasmic Reticulum-Phagy

Authors: Yong-Hui Zheng

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Viruses hijack host machineries to propagate and spread, which disrupts cellular homeostasis and activates various counteractive mechanisms. Infection of enveloped viruses is dependent on their fusion proteins, which bind to viral receptors to allow virus entry into cells. Fusion proteins are glycoproteins and expressed in the endoplasmic reticulum (ER) by hijacking the secretory pathway. Previously, we reported that Zaire ebolavirus (EBOV)-glycoprotein (GP) expression induces ER stress, and EBOV-GP is targeted by the calnexin cycle to macro-ER-phagy for degradation. We now report that expression of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2/SARS2)-spike (S) protein also causes ER stress, and its expression is strongly downregulated by mannosidase alpha class 1B member 1 (MAN1B1), a class I α-mannosidase from the ER. MAN1B1 co-localizes with SARS2-S in the ER, and its downregulation of SARS2-S is blocked by inhibitors targeting lysosomes and autophagy, but not proteasomes, indicating SARS2-S degradation by autolysosomes. Notably, the SARS2-S degradation does not require the core autophagy machinery including ATG3, ATG5, ATG7, and phosphatidylinositol 3-kinase catalytic subunit type 3 (PI3KC3)/vacuolar protein sorting 34 (VPS34), and instead, it requires Beclin 1 (BECN1), a core component in the PI3KC3 complex. In addition, MAN1B1 does not trigger SARS2-S polyubiquitination, and consistently, the SARS2-S degradation does not require the autophagy receptor sequestosome 1 (SQSTM1)/p62. MAN1B1 also downregulates EBOV-GP similarly, but this degradation does not require BECN1. Collectively, we conclude that MAN1B1 downregulates viral fusions by micro-ER-phagy, and importantly, we have identified BECN1-dependent and BECN1-independent mechanisms for micro-ER-phagy.

Keywords: Micro-ER-phagy, reticulophagy, fusion proteins, ER stress

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