Search results for: snp markers
Commenced in January 2007
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Edition: International
Paper Count: 324

Search results for: snp markers

84 Effects of Acupuncture Treatment in Gait Parameters in Parkinson's Disease

Authors: Catarina Isabel Ramos Pereira, Jorge Machado, Begona Alonso Criado, Maria João Santos

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Introduction: Gait disorders are one of the symptoms that have severe implications on the quality of life in Parkinson's disease (PD). Currently, there is no therapy to reverse or treat this condition. None of the drugs used in conventional medical treatment is entirely efficient, and all have a high incidence of side effects. Acupuncture treatment is believed to improve motor ability, but there is still little scientific evidence in individuals with PD. Aim: The aim of the study is to investigate the acute effect of acupuncture on gait parameters in Parkinson's disease. Methods: This is a randomized and controlled crossover study. The same individual patient was part of both the experimental (real acupuncture) and control group (false acupuncture/sham), and the sequence was randomized. Gait parameters were measured at two different moments, before and after treatment, using four force platforms as well as the collection of 3D markers positions taken by 11 cameras. Images were quantitatively analyzed using Qualisys Track Manager software that let us extract data related to the quality of gait and balance. Seven patients with the diagnosis of Parkinson's disease were included in the study. Results: Statistically significant differences were found in gait speed (p = 0.016), gait cadence (p = 0.006), support base width (p = 0.0001), medio-lateral oscillation (p = 0.017), left-right step length (p = 0.0002), and stride length: right-right (p = 0.0000) and left-left (p = 0.0018), time of left support phase (p = 0.029), right support phase (p = 0.025) and double support phase (p = 0.015), between the initial and final moments for the experimental group. Differences in right-left stride length were found for both groups. Conclusion: Our results show that acupuncture could enhance gait in Parkinson's disease patients. Deep research involving a larger number of volunteers should be accomplished to validate these encouraging findings.

Keywords: acupuncture, traditional Chinese medicine, Parkinson's disease, gait

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83 Detection of Bcl2 Polymorphism in Patient with Hepatocellular carcinoma

Authors: Mohamed Abdel-Hamid, Olfat Gamil Shaker, Doha El-Sayed Ellakwa, Eman Fathy Abdel-Maksoud

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Introduction: Despite advances in the knowledge of the molecular virology of hepatitis C virus (HCV), the mechanisms of hepatocellular injury in HCV infection are not completely understood. Hepatitis C viral infection (HCV) influences the susceptibility to apoptosis. This could lead to insufficient antiviral immune response and persistent viral infection. Aim of this study: was to examine whether BCL-2 gene polymorphism at codon 43 (+127G/A or Ala43Thr) has an impact on development of hepatocellular carcinoma caused by chronic hepatitis C Egyptian patients. Subjects and Methods: The study included three groups; group 1: composing of 30 patients with hepatocellular carcinoma (HCC), group 2 composing of 30 patients with HCV, group 3 composing of 30 healthy subjects matching the same age and socioeconomic status were taken as a control group. Gene polymorphism of BCL2 (Ala43Thr) were evaluated by PCR-RFLP technique and measured for all patients and controls. Results: The summed 43Thr genotype was more frequent and statistically significant in HCC patients as compared to control group. This genotype of BCL2 gene may inhibit the programmed cell death which leads to disturbance in tissue and cells homeostasis and reduction in immune regulation. This result leads to viral replication and HCV persistence. Moreover, virus produces variety of mechanisms to block genes participated in apoptosis. This mechanism proves that HCV patients who have 43Thr genotype are more susceptible to HCC. Conclusion: The data suggest for the first time that the BCL2 polymorphism is associated with the susceptibility to HCC in Egyptian populations and might be used as molecular markers for evaluating HCC risk. This study clearly demonstrated that Chronic HCV exhibit a deregulation of apoptosis with the disease progression. This provides an insight into the pathogenesis of chronic HCV infection, and may contribute to the therapy.

Keywords: BCL2 gene, Hepatitis C Virus, Hepatocellular carcinoma, sensitivity, specificity, apoptosis

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82 Genetic Analysis of the Endangered Mangrove Species Avicennia Marina in Qatar Detected by Inter-Simple Sequence Repeat DNA Markers

Authors: Talaat Ahmed, Amna Babssail

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Mangroves are evergreen trees and grow along the coastal areas of Qatar. The largest and oldest area of mangroves can be found around Al-Thakhira and Al-Khor. Other mangrove areas originate from fairly recent plantings by the government, although unfortunately the picturesque mangrove lake in Al-Wakra has now been uprooted. Avicinnia marina is the predominant mangrove species found in the region. Mangroves protect and stabilize low lying coastal land, and provide protection and food sources for estuarine and coastal fishery food chains. They also serve as feeding, breeding and nursery grounds for a variety of fish, crustaceans, reptiles, birds and other wildlife. A total of 21 individuals of A. marina, representing seven diverse Natural and artificial populations, were sampled throughout its range in Qatar. Leaves from 2-3 randomly selected trees at each location were collected. The locations are as follows: Al-Rawis, Ras-Madpak, Fuwairt, Summaseima, Al-khour, AL-Mafjar and Zekreet. Total genomic DNA was extracted using commercial DNeasy Plant System (Qiagen, Inc., Valencia, CA) kit to be used for genetic diversity analysis. Total of 12 (Inter-Simple Sequence Repeat) ISSR primers were used to amplify DNA fragments using genomic DNA. The 12 ISSR primers amplified polymorphic bands among mangrove samples in different areas as well as within each area indicating the existing of variation within each area and among the different areas of mangrove in Qatar. The results could characterize Avicinnia marina populations exist in different areas of Qatar and establish DNA fingerprint documentations for mangrove population to be used in further studies. Moreover, existing of genetic variation within and among Avicinnia marina populations is a strong indication for the ability of such populations to adapt different environmental conditions in Qatar. This study could be a warning to save mangrove in Qatar and save the environment as well.

Keywords: DNA fingerprint, Avicinnia marina, genetic analysis, Qatar

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81 Safety Study of Intravenously Administered Human Cord Blood Stem Cells in the Treatment of Symptoms Related to Chronic Inflammation

Authors: Brian M. Mehling, Louis Quartararo, Marine Manvelyan, Paul Wang, Dong-Cheng Wu

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Numerous investigations suggest that Mesenchymal Stem Cells (MSCs) in general represent a valuable tool for therapy of symptoms related to chronic inflammatory diseases. Blue Horizon Stem Cell Therapy Program is a leading provider of adult and children’s stem cell therapies. Uniquely we have safely and efficiently treated more than 600 patients with documenting each procedure. The purpose of our study is primarily to monitor the immune response in order to validate the safety of intravenous infusion of human umbilical cord blood derived MSCs (UC-MSCs), and secondly, to evaluate effects on biomarkers associated with chronic inflammation. Nine patients were treated for conditions associated with chronic inflammation and for the purpose of anti-aging. They have been given one intravenous infusion of UC-MSCs. Our study of blood test markers of 9 patients with chronic inflammation before and within three months after MSCs treatment demonstrates that there is no significant changes and MSCs treatment was safe for the patients. Analysis of different indicators of chronic inflammation and aging included in initial, 24-hours, two weeks and three months protocols showed that stem cell treatment was safe for the patients; there were no adverse reactions. Moreover data from follow up protocols demonstrates significant improvement in energy level, hair, nails growth and skin conditions. Intravenously administered UC-MSCs were safe and effective in the improvement of symptoms related to chronic inflammation. Further close monitoring and inclusion of more patients are necessary to fully characterize the advantages of UC-MSCs application in treatment of symptoms related to chronic inflammation.

Keywords: chronic inflammatory diseases, intravenous infusion, stem cell therapy, umbilical cord blood derived mesenchymal stem cells (UC-MSCs)

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80 Antioxidant Potential of Pomegranate Rind Extract Attenuates Pain, Inflammation and Bone Damage in Experimental Rats

Authors: Ritu Karwasra, Surender Singh

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Inflammation is an important physiological response of the body’s self-defense system that helps in eliminating and protecting organism from harmful stimuli and in tissue repair. It is a highly regulated protective response which helps in eliminating the initial cause of cell injury, and initiates the process of repair. The present study was designed to evaluate the ameliorative effect of pomegranate rind extract on pain and inflammation. Hydroalcoholic standardized rind extract of pomegranate at doses 50, 100 and 200 mg/kg and indomethacin (3 mg/kg) was tested against eddy’s hot plate induced thermal algesia, carrageenan (acute inflammation) and Complete Freund’s Adjuvant (chronic inflammation) induced models in Wistar rats. Parameters analyzed were inhibition of paw edema, measurement of joint diameter, levels of GSH, TBARS, SOD, TNF-α, radiographic imaging, tissue histology and synovial expression of pro-inflammatory cytokine receptor (TNF-R1). Radiological and light microscopical analysis were carried out to find out the bone damage in CFA-induced chronic inflammatory model. Findings of the present study revealed that pomegranate rind extract at a dose of 200 mg/kg caused a significant (p<0.05) reduction in paw swelling in both the inflammatory models. Nociceptive threshold was also significantly (p<0.05) improved. Immunohistochemical analysis of TNF-R1 in CFA-induced group showed elevated level, whereas reduction in level of TNF-R1 was observed in pomegranate (200 mg/kg). Henceforth, we might say that pomegranate produced a dose-dependent reduction in inflammation and pain along with the reduction in levels of oxidative stress markers and tissue histology, and the effect was found to be comparable to that of indomethacin. Thus, it can be concluded that pomegranate is a potential therapeutic target in the pathogenesis of inflammation and pain, and punicalagin is the major constituents found in rind extract might be responsible for the activity.

Keywords: carrageenan, inflammation, nociceptive-threshold, pomegranate, histopathology

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79 Developing Scaffolds for Tissue Regeneration using Low Temperature Plasma (LTP)

Authors: Komal Vig

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Cardiovascular disease (CVD)-related deaths occur in 17.3 million people globally each year, accounting for 30% of all deaths worldwide, with a predicted annual incidence of deaths to reach 23.3 million globally by 2030. Autologous bypass grafts remain an important therapeutic option for the treatment of CVD, but the poor quality of the donor patient’s blood vessels, the invasiveness of the resection surgery, and postoperative movement restrictions create issues. The present study is aimed to improve the endothelialization of intimal surface of graft by using low temperature plasma (LTP) to increase the cell attachment and proliferation. Polytetrafluoroethylene (PTFE) was treated with LTP. Air was used as the feed-gas, and the pressure in the plasma chamber was kept at 800 mTorr. Scaffolds were also modified with gelatin and collagen by dipping method. Human umbilical vein endothelial cells (HUVEC) were plated on the developed scaffolds, and cell proliferation was determined by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and by microscopy. mRNA expressions levels of different cell markers were investigated using quantitative real-time PCR (qPCR). XPS confirmed the introduction of oxygenated functionalities from LTP. HUVEC cells showed 80% seeding efficiency on the scaffold. Microscopic and MTT assays indicated increase in cell viability in LTP treated scaffolds, especially when treated with gelatin or collagen, compared to untreated scaffolds. Gene expression studies shows enhanced expression of cell adhesion marker Integrin- α 5 gene after LTP treatment. LTP treated scaffolds exhibited better cell proliferation and viability compared to untreated scaffolds. Protein treatment of scaffold increased cell proliferation. Based on our initial results, more scaffolds alternatives will be developed and investigated for cell growth and vascularization studies. Acknowledgments: This work is supported by the NSF EPSCoR RII-Track-1 Cooperative Agreement OIA-2148653.

Keywords: LTP, HUVEC cells, vascular graft, endothelialization

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78 Development of a New Method for the Evaluation of Heat Tolerant Wheat Genotypes for Genetic Studies and Wheat Breeding

Authors: Hameed Alsamadany, Nader Aryamanesh, Guijun Yan

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Heat is one of the major abiotic stresses limiting wheat production worldwide. To identify heat tolerant genotypes, a newly designed system involving a large plastic box holding many layers of filter papers positioned vertically with wheat seeds sown in between for the ease of screening large number of wheat geno types was developed and used to study heat tolerance. A collection of 499 wheat geno types were screened under heat stress (35ºC) and non-stress (25ºC) conditions using the new method. Compared with those under non-stress conditions, a substantial and very significant reduction in seedling length (SL) under heat stress was observed with an average reduction of 11.7 cm (P<0.01). A damage index (DI) of each geno type based on SL under the two temperatures was calculated and used to rank the genotypes. Three hexaploid geno types of Triticum aestivum [Perenjori (DI= -0.09), Pakistan W 20B (-0.18) and SST16 (-0.28)], all growing better at 35ºC than at 25ºC were identified as extremely heat tolerant (EHT). Two hexaploid genotypes of T. aestivum [Synthetic wheat (0.93) and Stiletto (0.92)] and two tetraploid genotypes of T. turgidum ssp dicoccoides [G3211 (0.98) and G3100 (0.93)] were identified as extremely heat susceptible (EHS). Another 14 geno types were classified as heat tolerant (HT) and 478 as heat susceptible (HS). Extremely heat tolerant and heat susceptible geno types were used to develop re combinant inbreeding line populations for genetic studies. Four major QTLs, HTI4D, HTI3B.1, HTI3B.2 and HTI3A located on wheat chromosomes 4D, 3B (x2) and 3A, explaining up to 34.67 %, 28.93 %, 13.46% % and 11.34% phenotypic variation, respectively, were detected. The four QTLs together accounted for 88.40% of the total phenotypic variation. Random wheat geno types possessing the four heat tolerant alleles performed significantly better under the heat condition than those lacking the heat tolerant alleles indicating the importance of the four QTLs in conferring heat tolerance in wheat. Molecular markers are being developed for marker assisted breeding of heat tolerant wheat.

Keywords: bread wheat, heat tolerance, screening, RILs, QTL mapping, association analysis

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77 Predicting Factors for Occurrence of Cardiac Arrest in Critical, Emergency and Urgency Patients in an Emergency Department

Authors: Angkrit Phitchayangkoon, Ar-Aishah Dadeh

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Background: A key aim of triage is to identify the patients with high risk of cardiac arrest because they require intensive monitoring, resuscitation facilities, and early intervention. We aimed to identify the predicting factors such as initial vital signs, serum pH, serum lactate level, initial capillary blood glucose, and Modified Early Warning Score (MEWS) which affect the occurrence of cardiac arrest in an emergency department (ED). Methods: We conducted a retrospective data review of ED patients in an emergency department (ED) from 1 August 2014 to 31 July 2016. Significant variables in univariate analysis were used to create a multivariate analysis. Differentiation of predicting factors between cardiac arrest patient and non-cardiac arrest patients for occurrence of cardiac arrest in an emergency department (ED) was the primary outcome. Results: The data of 527 non-trauma patients with Emergency Severity Index (ESI) 1-3 were collected. The factors found to have a significant association (P < 0.05) in the non-cardiac arrest group versus the cardiac arrest group at the ED were systolic BP (mean [IQR] 135 [114,158] vs 120 [90,140] mmHg), oxygen saturation (mean [IQR] 97 [89,98] vs 82.5 [78,95]%), GCS (mean [IQR] 15 [15,15] vs 11.5 [8.815]), normal sinus rhythm (mean 59.8 vs 30%), sinus tachycardia (mean 46.7 vs 21.7%), pH (mean [IQR] 7.4 [7.3,7.4] vs 7.2 [7,7.3]), serum lactate (mean [IQR] 2 [1.1,4.2] vs 7 [5,10.8]), and MEWS score (mean [IQR] 3 [2,5] vs 5 [3,6]). A multivariate analysis was then performed. After adjusting for multiple factors, ESI level 2 patients were more likely to have cardiac arrest in the ER compared with ESI 1 (odds ratio [OR], 1.66; P < 0.001). Furthermore, ESI 2 patients were more likely than ESI 1 patients to have cardiovascular disease (OR, 1.89; P = 0.01), heart rate < 55 (OR, 6.83; P = 0.18), SBP < 90 (OR, 3.41; P = 0.006), SpO2 < 94 (OR, 4.76; P = 0.012), sinus tachycardia (OR, 4.32; P = 0.002), lactate > 4 (OR, 10.66; P = < 0.001), and MEWS > 4 (OR, 4.86; P = 0.028). These factors remained predictive of cardiac arrest at the ED. Conclusion: The factors related to cardiac arrest in the ED are ESI 1 patients, ESI 2 patients, patients diagnosed with cardiovascular disease, SpO2 < 94, lactate > 4, and a MEWS > 4. These factors can be used as markers in the event of simultaneous arrival of many patients and can help as a pre-state for patients who have a tendency to develop cardiac arrest. The hemodynamic status and vital signs of these patients should be closely monitored. Early detection of potentially critical conditions to prevent critical medical intervention is mandatory.

Keywords: cardiac arrest, predicting factor, emergency department, emergency patient

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76 Curative Role of Bromoenol Lactone, an Inhibitor of Phospholipase A2 Enzyme, during Cigarette Smoke Condensate Induced Anomalies in Lung Epithelium

Authors: Subodh Kumar, Sanjeev Kumar Sharma, Gaurav Kaushik, Pramod Avti, Phulen Sarma, Bikash Medhi, Krishan Lal Khanduja

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Background: It is well known that cigarette smoke is one of the causative factors in various lung diseases especially cancer. Carcinogens and oxidant molecules present in cigarette smoke not only damage the cellular constituents (lipids, proteins, DNA) but may also regulate the molecular pathways involved in inflammation and cancer. Continuous oxidative stress caused by the constituents of cigarette smoke leads to higher PhospholipaseA₂ (PLA₂) activity, resulting in elevated levels of secondary metabolites whose role is well defined in cancer. To reduce the burden of chronic inflammation as well as oxidative stress, and higher levels of secondary metabolites, we checked the curative potential of PLA₂ inhibitor Bromoenol Lactone (BEL) during continuous exposure of cigarette smoke condensate (CSC). Aim: To check the therapeutic potential of Bromoenol Lactone (BEL), an inhibitor of PhospholipaseA₂s, in pathways of CSC-induced changes in type I and type II alveolar epithelial cells. Methods: Effect of BEL on CSC-induced PLA2 activity were checked using colorimetric assay, cellular toxicity using cell viability assay, membrane integrity using fluorescein di-acetate (FDA) uptake assay, reactive oxygen species (ROS) levels and apoptosis markers through flow cytometry, and cellular regulation using MAPKinases levels, in lung epithelium. Results: BEL significantly mimicked CSC-induced PLA₂ activity, ROS levels, apoptosis, and kinases level whereas improved cellular viability and membrane integrity. Conclusions: Current observations revealed that BEL may be a potential therapeutic agent during Cigarette smoke-induced anomalies in lung epithelium.

Keywords: cigarette smoke condensate, phospholipase A₂, oxidative stress, alveolar epithelium, bromoenol lactone

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75 Derivation of Human NK Cells from T Cell-Derived Induced Pluripotent Stem Cells Using Xenogeneic Serum-Free and Feeder Cell-Free Culture System

Authors: Aliya Sekenova, Vyacheslav Ogay

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The derivation of human induced pluripotent stem cells (iPSCs) from somatic cells by direct reprogramming opens wide perspectives in the regenerative medicine. It means the possibility to develop the personal and, consequently, any immunologically compatible cells for applications in cell-based therapy. The purpose of our study was to develop the technology for the production of NK cells from T cell-derived induced pluripotent stem cells (TiPSCs) for subsequent application in adoptive cancer immunotherapy. Methods: In this study iPSCs were derived from peripheral blood T cells using Sendai virus vectors expressing Oct4, Sox2, Klf4 and c-Myc. Pluripotent characteristics of TiPSCs were examined and confirmed with alkaline phosphatase staining, immunocytochemistry and RT-PCR analysis. For NK cell differentiation, embryoid bodies (EB) formed from (TiPSCs) were cultured in xenogeneic serum-free medium containing human serum, IL-3, IL-7, IL-15, SCF, FLT3L without using M210-B4 and AFT-024 stromal feeder cells. After differentiation, NK cells were characterized with immunofluorescence analysis, flow cytometry and cytotoxicity assay. Results: Here, we for the first time demonstrate that TiPSCs can effectively differentiate into functionally active NK cells without M210-B4 and AFT-024 xenogeneic stroma cells. Immunofluorescence and flow cytometry analysis showed that EB-derived cells can differentiate into a homogeneous population of NK cell expressing high levels of CD56, CD45 and CD16 specific markers. Moreover, these cells significantly express killing activation receptors such as NKp44 and NKp46. In the comparative analysis, we observed that NK cells derived using feeder-free culture system have more high killing activity against K-562 tumor cells, than NK cells derived by feeder-dependent method. Thus, we think that our obtained data will be useful for the development of large-scale production of NK cells for translation into cancer immunotherapy.

Keywords: induced pluripotent stem cells, NK cells, T cells, cell diffentiation, feeder cell-free culture system

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74 Culture of Human Mesenchymal Stem Cells Culture in Xeno-Free Serum-Free Culture Conditions on Laminin-521

Authors: Halima Albalushi, Mohadese Boroojerdi, Murtadha Alkhabori

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Introduction: Maintenance of stem cell properties during culture necessitates the recreation of the natural cell niche. Studies reported the promising outcome of mesenchymal stem cells (MSC) properties maintenance after using extracellular matrix such as CELLstart™, which is the recommended coating material for stem cells cultured in serum-free and xeno-free conditions. Laminin-521 is known as a crucial adhesion protein, which is found in natural stem cell niche, and plays an important role in facilitating the maintenance of self-renewal, pluripotency, standard morphology, and karyotype of human pluripotent stem cells (PSCs). The aim of this study is to investigate the effects of Laminin-521 on human umbilical cord-derived mesenchymal stem cells (UC-MSC) characteristics as a step toward clinical application. Methods: Human MSC were isolated from the umbilical cord via the explant method. Umbilical cord-derived-MSC were cultured in serum-free and xeno-free conditions in the presence of Laminin-521 for six passages. Cultured cells were evaluated by morphology and expansion index for each passage. Phenotypic characterization of UC-MSCs cultured on Laminin-521 was evaluated by assessment of cell surface markers. Results: Umbilical cord derived-MSCs formed small colonies and expanded as a homogeneous monolayer when cultured on Laminin-521. Umbilical cord derived-MSCs reached confluence after 4 days in culture. No statistically significant difference was detected in all passages when comparing the expansion index of UC-MSCs cultured on LN-521 and CELLstart™. Phenotypic characterization of UC-MSCs cultured on LN-521 using flow cytometry revealed positive expression of CD73, CD90, CD105 and negative expression of CD34, CD45, CD19, CD14 and HLA-DR.Conclusion: Laminin-521 is comparable to CELLstart™ in supporting UC-MSCs expansion and maintaining their characteristics during culture in xeno-free and serum-free culture conditions.

Keywords: mesenchymal stem cells, culture, laminin-521, xeno-free serum-free

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73 Effects of Pterostilbene in Brown Adipose Tissue from Obese Rats

Authors: Leixuri Aguirre, Iñaki Milton-Laskibar, Elizabeth Hijona, Luis Bujanda, Agnes M. Rimando, Maria P. Portillo

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Introduction: In recent years great attention has been paid by scientific community to phenolic compounds as active biomolecules naturally present in foodstuffs due to their beneficial effects on health. Pterostilbene is a resveratrol dimethylether derivative which shows higher biodisponibility. Objective. To analyze the effects of two doses of pterostilbene on several markers of thermogenic capacity in a model of genetic obesity, which shows reduced thermogenesis. Methods: The experiment was conducted with thirty Zucker (fa/fa) rats that were distributed in 3 experimental groups, the control group and two groups orally administered with pterostilbene at 15 and 30 mg/kg body weight/day for 6 weeks. Gene expression of Ucp1, Pgc-1α, Cpt1b, Pparα, Nfr1, Tfam and Cox-2 were assessed by RT-PCR, protein expression of UCP1 and GLUT4 by western blot and enzyme activity of carnitine palmitoyl transferase 1b and citrate synthase by spectrophotometry in interscapular brown adipose tissue (iBAT). Statistical analysis was performed by using one way ANOVA and Newman-Keuls as post-hoc test. Results: Pterostilbene did not change gene expression of Pgc-1α. However, significant increases were found in the expression of Ucp1, Pparα, Nfr-1 and Cox-2. Protein expression of UCP1 and GLUT4 was increased in animals treated with pterostilbene, as well as the activities of CPT-1b and CS. These effects were observed with both doses of pterostilbene, without differences between them. Conclusions: These results show that pterostilbene increases thermogenic and oxidative capacity of brown adipose tissue in obese rats. Whether these effects effectively contribute to the anti-obesity properties of these compound needs further research. Acknowledgments: MINECO-FEDER (AGL2015-65719-R), Basque Government (IT-572-13), University of the Basque Country (ELDUNANOTEK UFI11/32), Institut of Health Carlos III (CIBERobn). Iñaki Milton is a fellowship from the Basque Government.

Keywords: brown adipose tissue, pterostilbene, thermogenesis, uncoupling protein 1

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72 The Modulation of Health and Inflammatory Status in Young Pigs by Grape Waste Enriched in Polyphenols

Authors: Gina Cecilia Pistol, Loredana Calin, Mariana Stancu, Veronica Chedea, Ionelia Taranu

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Inflammatory-associated diseases have an increased trend in the past decades. The pharmacological strategies aimed to treat these inflammatory diseases are very expensive and with non-beneficial results. The current trend is to find alternative strategies to counteract or to control inflammatory component of diseases. The grape by-products either seeds or pomace are rich in bioactive compounds (e.g. polyphenols) which may be beneficial in prevention of inflammation associated with cancer progression and other pathologies with inflammatory component. The in vivo models are very useful for studying the immune and inflammatory status. The domestic pig (Sus scrofa domesticus) is related to human from anatomic and physiologic point of view, representing a feasible model for studying the human inflammatory pathologies. Starting from these data, we evaluated the effect of a diet containing 5% grape seed cakes (GS) on piglets blood biochemical parameters and immune pro- and anti-inflammatory biomarkers (IL-1 beta, IL-8, TNF-alpha, IL-6, IFN-gamma, IL-10, IL-4) in spleen and lymph nodes. 12 weaned piglets were fed for 30 days with a control diet or an experimental diet containing 5% GS. At the end of trial, plasma and tissue samples (spleen and lymph nodes) were collected and the biochemical and inflammatory markers were analysed by using biochemistry analyser and ELISA techniques. Our results showed that diet included 5% GS did not influence the health status determined by plasma biochemical parameters. Only a tendency for a slight increase of the biochemical parameters associated with energetic profile (glucose, cholesterol, triglycerides) was observed. Also, GS diet had no effect on pro- and anti-inflammatory cytokines content in spleen and lymph nodes tissue. Further experiments are needed in order to investigate other rate of dietary inclusion which could provide more evidence about the effect of grape bioactive compounds on pigs used as animal model.

Keywords: animal model, inflammation, grape seed by-product, immune organs

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

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

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

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

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70 The h3r Antagonist E159 Alleviates Neuroinflammation and Autistic-Like Phenotypes in BTBR T+ tf/J Mouse Model of Autism

Authors: Shilu Deepa Thomas, P. Jayaprakash, Dorota Łazewska, Katarzyna Kieć-Kononowicz, B. Sadek

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A large body of evidence suggests the involvement of cognitive impairment, increased levels of inflammation and oxidative stress in the pathogenesis of autism spectrum disorder (ASD). ASD commonly coexists with psychiatric conditions like anxiety and cognitive challenges, and individuals with ASD exhibit significant levels of inflammation and immune system dysregulation. Previous Studies have identified elevated levels of pro-inflammatory markers such as IL-1β, IL-6, IL-2 and TNF-α, particularly in young children with ASD. The current therapeutic options for ASD show limited effectiveness, signifying the importance of exploring an efficient drugs to address the core symptoms. The role of histamine H3 receptors (H3Rs) in memory and the prospective role of H3R antagonists in pharmacological control of neurodegenerative disorders, e.g., ASD, is well-accepted. Hence, the effects of chronic systemic administration of H3R antagonist E159 on autistic-like repetitive behaviors, social deficits, memory and anxiety parameters, as well as neuroinflammation in Black and Tan BRachyury (BTBR) mice, were evaluated using Y maze, Barnes maze, self-grooming, open field and three chamber social test. E159 (2.5, 5 and 10 mg/kg, i.p.) dose-dependently ameliorated repetitive and compulsive behaviors by reducing the increased time spent in self-grooming and improved reduced spontaneous alternation in BTBR mice. Moreover, treatment with E159 attenuated disturbed anxiety levels and social deficits in tested male BTBR mice. Furthermore, E159 attenuated oxidative stress by significantly increasing GSH, CAT, and SOD and decreasing the increased levels of MDA in the cerebellum as well as the hippocampus. In addition, E159 decreased the elevated levels of proinflammatory cytokines (tumor necrosis factor (TNF-α), interleukin-1β (IL-1β), and IL-6). The observed results show that H3R antagonists like E159 may represent a promising novel pharmacological strategy for the future treatment of ASD.

Keywords: histamine H3 receptors, antagonist E159, autism, behaviors, mice

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69 Tinder as a Queer Identity Building Tool: An Ethnographic Study of How Dating Apps Are Used as Identity Markers among Queer Women in Bologna, Italy

Authors: Lovis Ingrid Hakala

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This paper provides an example of how the dating app Tinder can be used as a tool in queer research and what the use of the app among lesbians can tell us about the production of particular kinds of lesbian identities paired with technology. This study seeks to understand the ways in which the app is understood by lesbian women and how it is brought into their understandings of what being a lesbian means. Dating apps are becoming increasingly common as tools for meeting a potential partner and research has shown how these apps gain new meaning depending on the context. This paper will show both how Tinder has gained a particular meaning in a lesbian context in Bologna, and also how the use of Tinder produces specific ideas of what lesbians are and how they act. While doing ethnographic fieldwork among young lesbians (20-30 years old) in Bologna in the autumn of 2017, as a part of the research methodology, Tinder was used together with interviews and participant observation. By signing up on Tinder and contacting potential informants on the app the aim was to reach women who were not necessarily involved in the queer activist spaces in the city and who might thus provide a different perspective than their activist counterparts. Using Tinder as a tool in research proved to be a useful methodology both when it came to contacting potential informants as well as in gaining a better understanding for queer identity building, and the role Tinder played in defining lesbianism in Bologna. The app itself was often deemed useless, and informants often complained about the lack of women on the app who had their profile showing an interest in women. Apart from being away for the interlocutors to get in touch with potential romantic or sexual partners, the app did provide a concrete object around which to define particular lesbian behavior. Lesbians were described to be slow at answering and not being very eager to meet. This placed lesbian women, in contrast to gay men who were seen as doing the opposite, using dating apps a lot and be quick to propose meetings. Tinder among lesbians in Bologna thus proved to have a dual function as both a way of creating connections between queer women but also as a tool to define what a lesbian is. This paper provides insight into the ways in which dating apps provides a community and identity building function and how a certain behavior on said apps can be a part of defining identity. This work stands out among other literature on dating apps since it does not only focus on the methodological aspects of using Tinder as a tool in research but also provides an insight in how the way in which individuals use the app comes to add to an understanding of what a lesbian is.

Keywords: dating apps, identity, Italy, lesbian, queer

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68 Characterization of Transmembrane Proteins with Five Alpha-Helical Regions

Authors: Misty Attwood, Helgi Schioth

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Transmembrane proteins are important components in many essential cell processes such as signal transduction, cell-cell signalling, transport of solutes, structural adhesion activities, and protein trafficking. Due to their involvement in diverse critical activities, transmembrane proteins are implicated in different disease pathways and hence are the focus of intense interest in understanding functional activities, their pathogenesis in disease, and their potential as pharmaceutical targets. Further, as the structure and function of proteins are correlated, investigating a group of proteins with the same tertiary structure, i.e., the same number of transmembrane regions, may give understanding about their functional roles and potential as therapeutic targets. In this in silico bioinformatics analysis, we identify and comprehensively characterize the previously unstudied group of proteins with five transmembrane-spanning regions (5TM). We classify nearly 60 5TM proteins in which 31 are members of ten families that contain two or more family members and all members are predicted to contain the 5TM architecture. Furthermore, nine singlet proteins that contain the 5TM architecture without paralogues detected in humans were also identifying, indicating the evolution of single unique proteins with the 5TM structure. Interestingly, more than half of these proteins function in localization activities through movement or tethering of cell components and more than one-third are involved in transport activities, particularly in the mitochondria. Surprisingly, no receptor activity was identified within this family in sharp contrast with other TM families. Three major 5TM families were identified and include the Tweety family, which are pore-forming subunits of the swelling-dependent volume regulated anion channel in astrocytes; the sidoreflexin family that acts as mitochondrial amino acid transporters; and the Yip1 domain family engaged in vesicle budding and intra-Golgi transport. About 30% of the proteins have enhanced expression in the brain, liver, or testis. Importantly, 60% of these proteins are identified as cancer prognostic markers, where they are associated with clinical outcomes of various tumour types, indicating further investigation into the function and expression of these proteins is important. This study provides the first comprehensive analysis of proteins with 5TM regions and provides details of the unique characteristics and application in pharmaceutical development.

Keywords: 5TM, cancer prognostic marker, drug targets, transmembrane protein

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67 Serum Vitamin D and Carboxy-Terminal TelopeptideType I Collagen Levels: As Markers for Bone Health Affection in Patients Treated with Different Antiepileptic Drugs

Authors: Moetazza M. Al-Shafei, Hala Abdel Karim, Eitedal M. Daoud, Hassan Zaki Hassuna

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Epilepsy is a common neurological disorder affecting all age groups. It is one of the world's most prevalent non-communicable diseases. Increased evidence suggesting that long term usage of anti-epileptic drugs can have adverse effects on bone mineralization and bone molding .Aiming to study these effects and to give guide lines to support bone health through early intervention. From Neurology Out-Patient Clinic kaser Elaini University Hospital, 60 Patients were enrolled, 40 patients on antiepileptic drugs for at least two years and 20 controls matched with age and sex, epileptic but before starting treatment both chosen under specific criteria. Patients were divided into four groups, three groups with monotherapy treated with either Phynetoin, Valporic acid or Carbamazipine and fourth group treated with both Valporic acid and Carbamazipine. Estimation of serum Carboxy-Terminal Telopeptide of Type I- Collagen(ICTP) bone resorption marker, serum 25(OH )vit D3, calcium ,magnesium and phosphorus were done .Results showed that all patients on AED had significant low levels of 25(OH) vit D3 (p<0.001) ,with significant elevation of ICTP (P<0.05) versus controls. In group treated with Phynotoin highly significant elevation of (ICTP) marker and decrease of both serum 25(OH) vit D3 (P<0, 0001) and serum calcium(P<0.05)versus control. Double drug group showed significant decrease of serum 25(OH) vit D3 (P<0.0001) and decrease in Phosphorus (P<0.05) versus controls. Serum magnesium showed no significant differences between studied groups. We concluded that Anti- epileptic drugs appears to be an aggravating factor on bone mineralization ,so therapeutically it can be worth wile to supplement calcium and vitamin D even before initiation of antiepileptic therapy. ICTP marker can be used to evaluate change in bone resorption before and during AED therapy.

Keywords: antiepileptic drugs, bone minerals, carboxy teminal telopeptidetype-1-collagen bone resorption marker, vitamin D

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66 A Novel Application of CORDYCEPIN (Cordycepssinensis Extract): Maintaining Stem Cell Pluripotency and Improving iPS Generation Efficiency

Authors: Shih-Ping Liu, Cheng-Hsuan Chang, Yu-Chuen Huang, Shih-Yin Chen, Woei-Cherng Shyu

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Embryonic stem cells (ES) and induced pluripotnet stem cells (iPS) are both pluripotent stem cells. For mouse stem cells culture technology, leukemia inhibitory factor (LIF) was used to maintain the pluripotency of stem cells in vitro. However, LIF is an expensive reagent. The goal of this study was to find out a pure compound extracted from Chinese herbal medicine that could maintain stem cells pluripotency to replace LIF and improve the iPS generation efficiency. From 20 candidates traditional Chinese medicine we found that Cordycepsmilitaris triggered the up-regulation of stem cells activating genes (Oct4 and Sox2) expression levels in MEF cells. Cordycepin, a major active component of Cordycepsmilitaris, also could up-regulate Oct4 and Sox2 gene expression. Furthermore, we used ES and iPS cells and treated them with different concentrations of Cordycepin (replaced LIF in the culture medium) to test whether it was useful to maintain the pluripotency. The results showed higher expression levels of several stem cells markers in 10 μM Cordycepin-treated ES and iPS cells compared to controls that did not contain LIF, including alkaline phosphatase, SSEA1, and Nanog. Embryonic body formation and differentiation confirmed that 10 μM Cordycepin-containing medium was capable to maintain stem cells pluripotency after four times passages. For mechanism analysis, microarray analysis indicated extracellular matrix and Jak/Stat signaling pathway as the top two deregulated pathways. In ECM pathway, we determined that the integrin αVβ5 expression levels and phosphorylated Src levels increased after Cordycepin treatment. In addition, the phosphorylated Jak2 and phosphorylated Sat3 protein levels were increased after Cordycepin treatment and suppressed with the Jak2 inhibitor, AG490. The expression of cytokines associated with Jak2/Stat3 signaling pathway were also up-regulated by Q-PCR and ELISA assay. Lastly, we used Oct4-GFP MEF cells to test iPS generation efficiency following Cordycepin treatment. We observed that 10 Μm Cordycepin significantly increased the iPS generation efficiency in day 21. In conclusion, we demonstrated Cordycepin could maintain the pluripotency of stem cells through both of ECM and Jak2/Stat3 signaling pathway and improved iPS generation efficiency.

Keywords: cordycepin, iPS cells, Jak2/Stat3 signaling pathway, molecular biology

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65 The Effect of Foot Progression Angle on Human Lower Extremity

Authors: Sungpil Ha, Ju Yong Kang, Sangbaek Park, Seung-Ju Lee, Soo-Won Chae

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The growing number of obese patients in aging societies has led to an increase in the number of patients with knee medial osteoarthritis (OA). Artificial joint insertion is the most common treatment for knee medial OA. Surgery is effective for patients with serious arthritic symptoms, but it is costly and dangerous. It is also inappropriate way to prevent a disease as an early stage. Therefore Non-operative treatments such as toe-in gait are proposed recently. Toe-in gait is one of non-surgical interventions, which restrain the progression of arthritis and relieves pain by reducing knee adduction moment (KAM) to facilitate lateral distribution of load on to knee medial cartilage. Numerous studies have measured KAM in various foot progression angle (FPA), and KAM data could be obtained by motion analysis. However, variations in stress at knee cartilage could not be directly observed or evaluated by these experiments of measuring KAM. Therefore, this study applied motion analysis to major gait points (1st peak, mid –stance, 2nd peak) with regard to FPA, and to evaluate the effects of FPA on the human lower extremity, the finite element (FE) method was employed. Three types of gait analysis (toe-in, toe-out, baseline gait) were performed with markers placed at the lower extremity. Ground reaction forces (GRF) were obtained by the force plates. The forces associated with the major muscles were computed using GRF and marker trajectory data. MRI data provided by the Visible Human Project were used to develop a human lower extremity FE model. FE analyses for three types of gait simulations were performed based on the calculated muscle force and GRF. We observed the maximum stress point during toe-in gait was lower than the other types, by comparing the results of FE analyses at the 1st peak across gait types. This is the same as the trend exhibited by KAM, measured through motion analysis in other papers. This indicates that the progression of knee medial OA could be suppressed by adopting toe-in gait. This study integrated motion analysis with FE analysis. One advantage of this method is that re-modeling is not required even with changes in posture. Therefore another type of gait simulation or various motions of lower extremity can be easily analyzed using this method.

Keywords: finite element analysis, gait analysis, human model, motion capture

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64 Carbendazim Toxicity and Ameliorative Effect of Vitamin E in African Giant Rats

Authors: A. O. Omonona, T. A. Jarikre

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Increase specialization in agriculture and use of pesticides may inadvertently cause ecosystem degradation and eventually loss of biodiversity. The populations of numerous wildlife species have undergone a precipitous decline. Many of these problems have been attributed directly to habitat loss and over exploitation resulting from unregulated pesticide uses. Carbendazim a broad spectrum benzimidazole fungicide and a metabolite of benomyl, is used to control plant disease in cereals and fruit. The effect of carbendazim exposure and the ameliorative effect of tocopherol (vitamin E) were assessed on African giant rat AGR. Hematological, biochemical and histological changes were used to determine the health condition of the animals exposed to pesticide. Sixteen AGR were stabilized, weighed and then divided into four experimental groups (A to D). Two groups were pretreated with vitamin. Group A was exposed to carbendazim only, B- carbendazim + vitamin, C- vitamin only, and D- blank (control). Packed cell volume PCV was estimated by the microhematocrit method, Leucocyte and Platelet counts were determined using the hemocytometric method. Cholinesterase (AchE) and markers of oxidative stress were quantified, and tissue changes examined microscopically. There were no behavioral changes observed in the animals, but there was a decrease in body weight and abortion after 23 days of exposure to carbendazim. There was significant differences in the packed cell volume, the hemoglobin concentration and the red blood cell counts (p < 0.05). The increases in malonyl aldehyde MDA was significant (p < 0.05) in the pesticide intoxicated rats compared to control. Vitamin E supplementation reduced MDA level significantly (p < 0.05). There was a sharp remarkable decrease in acetylcholinesterase levels in the pesticide intoxicated rats (p < 0.05). Vitamin E supplementation normalise the AchE levels comparable to that in control. Grossly, the vital organs appeared normal in the pesticide exposed and control groups except moderate pulmonary congestion. Microscopically, there was severe diffuse hepatocellular swelling in carbendazim exposed group. The severity of hepatocellular injury was reduced in the rats with vitamin E. This study ascertained the toxic effect of carbendazim and antioxidative properties of vitamins in the Africa giant rat.

Keywords: African giant rat, antioxidant, carbendazim, pesticides, toxicity

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63 Food Losses Reducing by Extending the Minimum Durability Date of Thermally Processed Products

Authors: Dorota Zielińska, Monika Trząskowska, Anna Łepecka, Katarzyna Neffe-Skocińska, Beata Bilska, Marzena Tomaszewska, Danuta Kołożyn-Krajewska

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Minimum durability date (MDD) labeled food is known to have a long shelf life. A properly stored or transported food retains its physical, chemical, microbiological, and sensory properties up to MDD. The aim of the study was to assess the sensory quality and microbiological safety of selected thermally processed products,i.e., mayonnaise, jam, and canned tuna within and after MDD. The scope of the study was to determine the markers of microbiological quality, i.e., the total viable count (TVC), the Enterobacteriaceae count and the total yeast and mold (TYMC) count on the last day of MDD and after 1 and 3 months of storage, after the MDD expired. In addition, the presence of Salmonella and Listeria monocytogenes was examined on the last day of MDD. The sensory quality of products was assessed by quantitative descriptive analysis (QDA), the intensity of differentiators (quality features), and overall quality were defined and determined. It was found that during three months storage of tested food products, after the MDD expired, the microbiological quality slightly decreased, however, regardless of the tested sample, TVC was at the level of <3 log cfu/g, similarly, the Enterobacretiaceae, what indicates the good microbiological quality of the tested foods. The TYMC increased during storage but did not exceed 2 logs cfu/g of product. Salmonella and Listeria monocytogenes were not found in any of the tested food samples. The sensory quality of mayonnaise negatively changed during storage. After three months from the expiry of MDD, a decrease in the "fat" and "egg" taste and aroma intensity, as well as the "density" were found. The "sour" taste intensity of blueberry jam after three months of storage was slightly higher, compared to the jam tested on the last day of MDD, without affecting the overall quality. In the case of tuna samples, an increase in the "fishy" taste and aroma intensity was observed during storage, and the overall quality did not change. Tested thermally processed products (mayonnaise, jam, and canned tuna) were characterized by good microbiological and sensory quality on the last day of MDD, as well as after three months of storage under conditions recommended by the producer. These findings indicate the possibility of reducing food losses by extending or completely abolishing the MDD of selected thermal processed food products.

Keywords: food wastes, food quality and safety, mayonnaise, jam, tuna

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62 Analysis of Anti-Tuberculosis Immune Response Induced in Lungs by Intranasal Immunization with Mycobacterium indicus pranii

Authors: Ananya Gupta, Sangeeta Bhaskar

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Mycobacterium indicus pranii (MIP) is a saprophytic mycobacterium. It is a predecessor of M. avium complex (MAC). Whole genome analysis and growth kinetics studies have placed MIP in between pathogenic and non-pathogenic species. It shares significant antigenic repertoire with M. tuberculosis and have unique immunomodulatory properties. MIP provides better protection than BCG against pulmonary tuberculosis in animal models. Immunization with MIP by aerosol route provides significantly higher protection as compared to immunization by subcutaneous (s.c.) route. However, mechanism behind differential protection has not been studied. In this study, using mice model we have evaluated and compared the M.tb specific immune response in lung compartments (airway lumen / lung interstitium) as well as spleen following MIP immunization via nasal (i.n.) and s.c. route. MIP i.n. vaccination resulted in increased seeding of memory T cells (CD4+ and CD8+ T-cells) in the airway lumen. Frequency of CD4+ T cells expressing Th1 migratory marker (CXCR3) and activation marker (CD69) were also high in airway lumen of MIP i.n. group. Significantly high ex vivo secretion of cytokines- IFN-, IL-12, IL-17 and TNF- from cells of airway luminal spaces provides evidence of antigen-specific lung immune response, besides generating systemic immunity comparable to MIP s.c. group. Analysis of T cell response on per cell basis revealed that antigen specific T-cells of MIP i.n. group were functionally superior as higher percentage of these cells simultaneously secreted IFN-gamma, IL-2 and TNF-alpha cytokines as compared to MIP s.c. group. T-cells secreting more than one of the cytokines simultaneously are believed to have robust effector response and crucial for protection, compared with single cytokine secreting T-cells. Adoptive transfer of airway luminal T-cells from MIP i.n. group into trachea of naive B6 mice revealed that MIP induced CD8 T-cells play crucial role in providing long term protection. Thus the study demonstrates that MIP intranasal vaccination induces M.tb specific memory T-cells in the airway lumen that results in an early and robust recall response against M.tb infection.

Keywords: airway lumen, Mycobacterium indicus pranii, Th1 migratory markers, vaccination

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61 Genetic Diversity of Sugar Beet Pollinators

Authors: Ksenija Taški-Ajdukovic, Nevena Nagl, Živko Ćurčić, Dario Danojević

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Information about genetic diversity of sugar beet parental populations is of a great importance for hybrid breeding programs. The aim of this research was to evaluate genetic diversity among and within populations and lines of diploid sugar beet pollinators, by using SSR markers. As plant material were used eight pollinators originating from three USDA-ARS breeding programs and four pollinators from Institute of Field and Vegetable Crops, Novi Sad. Depending on the presence of self-fertility gene, the pollinators were divided into three groups: autofertile (inbred lines), autosterile (open-pollinating populations), and group with partial presence of autofertility gene. A total of 40 SSR primers were screened, out of which 34 were selected for the analysis of genetic diversity. A total of 129 different alleles were obtained with mean value 3.2 alleles per SSR primer. According to the results of genetic variability assessment the number and percentage of polymorphic loci was the maximal in pollinators NS1 and tester cms2 while effective number of alleles, expected heterozygosis and Shannon’s index was highest in pollinator EL0204. Analysis of molecular variance (AMOVA) showed that 77.34% of the total genetic variation was attributed to intra-varietal variance. Correspondence analysis results were very similar to grouping by neighbor-joining algorithm. Number of groups was smaller by one, because correspondence analysis merged IFVCNS pollinators with CZ25 into one group. Pollinators FC220, FC221 and C 51 were in the next group, while self-fertile pollinators CR10 and C930-35 from USDA-Salinas were separated. On another branch were self-sterile pollinators ЕL0204 and ЕL53 from USDA-East Lansing. Sterile testers cms1 and cms2 formed separate group. The presented results confirmed that SSR analysis can be successfully used in estimation of genetic diversity within and among sugar beet populations. Since the tested pollinator differed considering the presence of self-fertility gene, their heterozygosity differed as well. It was lower in genotypes with fixed self-fertility genes. Since the most of tested populations were open-pollinated, which rarely self-pollinate, high variability within the populations was expected. Cluster analysis grouped populations according to their origin.

Keywords: auto fertility, genetic diversity, pollinator, SSR, sugar beet

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60 Histological and Morphometric Studies of the Liver of Goats Aborted

Authors: Toumi Farah, Charallah Salima

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In the Algerian Sahara, goat farming is predominant, and it’s associated with other types of breeding, particularly camel and sheep; it also constitutes a significant proportion of breeding exclusively goat. This Saharan goat is a small ruminant with a black dress with white’s spots, hanging ears, and a coat more or less long. It is known for its hardiness and resistance to adverse conditions of arid zones and its perfect ecophysiological adaptation to harsh environmental conditions. However, pregnancy alterations, particularly abortion, degrade its productivity and cause economic losses, having both direct and indirect effects on animal production, like the costs of veterinary interventions and the reconstitution of livestock. The purpose of this work is to study the histological aspect of the liver of goats’ aborted living under nomadic herds in the region of Béni-Abbès (30° 7' N, 2° 10 'O). The organs were collected in physiological serum, rinsed, and then fixed with formaldehyde (37°, diluted at 10%). After that, these samples were processed for a topographic study. The morphometric study of the liver was performed by using an image analysis and processing software "Image J"; the various measurements obtained are intended to specify the supposed stage of development according to the body weight. The histological structure of the liver shows that the hepatic parenchyma consists of vascular conjunctive spaces surrounded by Glisson’s capsule. The sinusoids and hepatic portal vein are full of red blood cells, representing sinusoidal congestion and a thrombosed vein. At high magnification, the blood vessels show the presence of vascular thrombosis and haemorrhage in some areas of the hepatic parenchyma. Morphometric analysis shows that the number of liver parenchymal cells and the diameter of liver vessels vary according to the stage of development. The results obtained will provide details of the anatomical and cellular elements that can be used in the diagnosis of early or late abortion and late embryonic death. It would be interesting to find, by immunohistochemistry, some inflammatory markers useful for monitoring the progress of pregnancy and bioindicators of fetomaternal distress.

Keywords: aborting goat, arid zone, liver, histopathology

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59 Habitat Suitability, Genetic Diversity and Population Structure of Two Sympatric Fruit Bat Species Reveal the Need of an Urgent Conservation Action

Authors: Mohamed Thani Ibouroi, Ali Cheha, Claudine Montgelard, Veronique Arnal, Dawiyat Massoudi, Guillelme Astruc, Said Ali Ousseni Dhurham, Aurelien Besnard

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The Livingstone's flying fox (Pteropus livingstonii) and the Comorian fruit bat (P.seychellensis comorensis) are two endemic fruit bat species among the mostly threatened animals of the Comoros archipelagos. Despite their role as important ecosystem service providers like all flying fox species as pollinators and seed dispersers, little is known about their ecologies, population genetics and structures making difficult the development of evidence-based conservation strategies. In this study, we assess spatial distribution and ecological niche of both species using Species Distribution Modeling (SDM) based on the recent Ensemble of Small Models (ESMs) approach using presence-only data. Population structure and genetic diversity of the two species were assessed using both mitochondrial and microsatellite markers based on non-invasive genetic samples. Our ESMs highlight a clear niche partitioning of the two sympatric species. Livingstone’s flying fox has a very limited distribution, restricted on steep slope of natural forests at high elevation. On the contrary, the Comorian fruit bat has a relatively large geographic range spread over low elevations in farmlands and villages. Our genetic analysis shows a low genetic diversity for both fruit bats species. They also show that the Livingstone’s flying fox population of the two islands were genetically isolated while no evidence of genetic differentiation was detected for the Comorian fruit bats between islands. Our results support the idea that natural habitat loss, especially the natural forest loss and fragmentation are the important factors impacting the distribution of the Livingstone’s flying fox by limiting its foraging area and reducing its potential roosting sites. On the contrary, the Comorian fruit bats seem to be favored by human activities probably because its diets are less specialized. By this study, we concluded that the Livingstone’s flying fox species and its habitat are of high priority in term of conservation at the Comoros archipelagos scale.

Keywords: Comoros islands, ecological niche, habitat loss, population genetics, fruit bats, conservation biology

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58 Value of FOXP3 Expression in Prediction of Neoadjuvant Chemotherapy Effect in Triple Negative Breast Cancer

Authors: Badawia Ibrahim, Iman Hussein, Samar El Sheikh, Fatma Abou Elkasem, Hazem Abo Ismael

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Background: Response of breast carcinoma to neoadjuvant chemotherapy (NAC) varies regarding many factors including hormonal receptor status. Breast cancer is a heterogenous disease with different outcomes, hence a need arises for new markers predicting the outcome of NAC especially for the triple negative group when estrogen, progesterone receptors and Her2/neu are negative. FOXP3 is a promising target with unclear role. Aim: To examine the value of FOXP3 expression in locally advanced triple negative breast cancer tumoral cells as well as tumor infiltrating lymphocytes (TILs) and to elucidate its relation to the extent of NAC response. Material and Methods: Forty five cases of immunohistochemically confirmed to be triple negative breast carcinoma were evaluated for NAC (Doxorubicin, Cyclophosphamide AC x 4 cycles + Paclitaxel x 12 weeks, patients with ejection fraction less than 60% received Taxotere or Cyclophosphamide, Methotrexate, Fluorouracil CMF) response in both tumour and lymph nodes status according to Miller & Payne's and Sataloff's systems. FOXP3 expression in tumor as well as TILs evaluated in the pretherapy biopsies was correlated with NAC response in breast tumor and lymph nodes as well as other clinicopathological factors. Results: Breast tumour cells showed FOXP3 positive cytoplasmic expression in (42%) of cases. High FOXP3 expression percentage was detected in (47%) of cases. High infiltration by FOXP3+TILs was detected in (49%) of cases. Positive FOXP3 expression was associated with negative lymph node metastasis. High FOXP3 expression percentage and high infiltration by FOXP3+TILs were significantly associated with complete therapy response in axillary lymph nodes. High FOXP3 expression in tumour cells was associated with high infiltration by FOXP3+TILs. Conclusion: This result may provide evidence that FOXP3 marker is a good prognostic and predictive marker for triple negative breast cancer (TNBC) indicated for neoadjuvant chemotherapy and can be used for stratifications of TNBC cases indicated for NAC. As well, this study confirmed the fact that the tumour cells and the surrounding microenvironment interact with each other and the tumour microenvironment can influence the treatment outcomes of TNBC.

Keywords: breast cancer, FOXP3 expression, prediction of neoadjuvant chemotherapy effect, triple negative

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57 Literary Words of Foreign Origin as Social Markers in Jeffrey Archer's Novels Speech Portrayals

Authors: Tatiana Ivushkina

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The paper is aimed at studying the use of literary words of foreign origin in modern fiction from a sociolinguistic point of view, which presupposes establishing correlation between this category of words in a speech portrayal or narrative and a social status of the speaker, verifying that it bears social implications and serves as a social marker or index of socially privileged identity in the British literature of the 21-st century. To this end, there were selected literary words of foreign origin in context (60 contexts) and subjected to careful examination. The study is carried out on two novels by Jeffrey Archer – Not a Penny More, Not a Penny Less and A Prisoner of Birth – who, being a graduate from Oxford, represents socially privileged classes himself and gives a wide depiction of characters with different social backgrounds and statuses. The analysis of the novels enabled us to categorize the selected words into four relevant groups. The first represented by terms (commodity, debenture, recuperation, syringe, luminescence, umpire, etc.) serves to unambiguously indicate education, occupation, a field of knowledge in which a character is involved or a situation of communication. The second group is formed of words used in conjunction with their Germanic counterparts (perspiration – sweat, padre – priest, convivial – friendly) to contrast social position of the characters: literary words serving as social indices of upper class speakers whereas their synonyms of Germanic origin characterize middle or lower class speech portrayals. The third class of words comprises socially marked words (verbs, nouns, and adjectives), or U-words (the term first coined by Allan Ross and Nancy Mitford), the status acquired in the course of social history development (elegant, excellent, sophistication, authoritative, preposterous, etc.). The fourth includes words used in a humorous or ironic meaning to convey the narrator’s attitude to the characters or situation itself (ministrations, histrionic, etc.). Words of this group are perceived as 'alien', stylistically distant as they create incongruity between style and subject matter. Social implication of the selected words is enhanced by French words and phrases often accompanying them.

Keywords: British literature of the XXI century, literary words of foreign origin, social context, social meaning

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56 Bioreactor for Cell-Based Impedance Measuring with Diamond Coated Gold Interdigitated Electrodes

Authors: Roman Matejka, Vaclav Prochazka, Tibor Izak, Jana Stepanovska, Martina Travnickova, Alexander Kromka

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Cell-based impedance spectroscopy is suitable method for electrical monitoring of cell activity especially on substrates that cannot be easily inspected by optical microscope (without fluorescent markers) like decellularized tissues, nano-fibrous scaffold etc. Special sensor for this measurement was developed. This sensor consists of corning glass substrate with gold interdigitated electrodes covered with diamond layer. This diamond layer provides biocompatible non-conductive surface for cells. Also, a special PPFC flow cultivation chamber was developed. This chamber is able to fix sensor in place. The spring contacts are connecting sensor pads with external measuring device. Construction allows real-time live cell imaging. Combining with perfusion system allows medium circulation and generating shear stress stimulation. Experimental evaluation consist of several setups, including pure sensor without any coating and also collagen and fibrin coating was done. The Adipose derived stem cells (ASC) and Human umbilical vein endothelial cells (HUVEC) were seeded onto sensor in cultivation chamber. Then the chamber was installed into microscope system for live-cell imaging. The impedance measurement was utilized by vector impedance analyzer. The measured range was from 10 Hz to 40 kHz. These impedance measurements were correlated with live-cell microscopic imaging and immunofluorescent staining. Data analysis of measured signals showed response to cell adhesion of substrates, their proliferation and also change after shear stress stimulation which are important parameters during cultivation. Further experiments plan to use decellularized tissue as scaffold fixed on sensor. This kind of impedance sensor can provide feedback about cell culture conditions on opaque surfaces and scaffolds that can be used in tissue engineering in development artificial prostheses. This work was supported by the Ministry of Health, grants No. 15-29153A and 15-33018A.

Keywords: bio-impedance measuring, bioreactor, cell cultivation, diamond layer, gold interdigitated electrodes, tissue engineering

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55 Agro-Morphological Traits Based Genetic Diversity Analysis of ‘Ethiopian Dinich’ Plectranthus edulis (Vatke) Agnew Populations Collected from Diverse Agro-Ecologies in Ethiopia

Authors: Fekadu Gadissa, Kassahun Tesfaye, Kifle Dagne, Mulatu Geleta

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‘Ethiopian dinich’ also called ‘Ethiopian potato’ is one of the economically important ‘orphan’ edible tuber crops indigenous to Ethiopia. We evaluated the morphological and agronomic traits performances of 174 samples from Ethiopia at multiple locations using 12 qualitative and 16 quantitative traits, recorded at the correct growth stages. We observed several morphotypes and phenotypic variations for qualitative traits along with a wide range of mean performance values for all quantitative traits. Analysis of variance for each quantitative trait showed a highly significant (p<0.001) variation among the collections with eventually non-significant variation for environment-traits interaction for all but flower length. A comparatively high phenotypic and genotypic coefficient of variation was observed for plant height, days to flower initiation, days to 50% flowering and tuber number per hill. Moreover, the variability and coefficients of variation due to genotype-environment interaction was nearly zero for all the traits except flower length. High genotypic coefficients of variation coupled with a high estimate of broad sense heritability and high genetic advance as a percent of collection mean were obtained for tuber weight per hill, number of primary branches per plant, tuber number per hill and number of plants per hill. Association of tuber yield per hectare of land showed a large magnitude of positive phenotypic and genotypic correlation with those traits. Principal components analysis revealed 76% of the total variation for the first six principal axes with high factor loadings again from tuber number per hill, number of primary branches per plant and tuber weight. The collections were grouped into four clusters with the weak region (zone) of origin based pattern. In general, there is high genetic-based variability for ‘Ethiopian dinich’ improvement and conservation. DNA based markers are recommended for further genetic diversity estimation for use in breeding and conservation.

Keywords: agro-morphological traits, Ethiopian dinich, genetic diversity, variance components

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