Search results for: gene modification
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 2483

Search results for: gene modification

593 Optimization of an Electro-Submersible Pump for Crude Oil Extraction Processes

Authors: Deisy Becerra, Nicolas Rios, Miguel Asuaje

Abstract:

The Electrical Submersible Pump (ESP) is one of the most artificial lifting methods used in the last years, which consists of a serial arrangement of centrifugal pumps. One of the main concerns when handling crude oil is the formation of O/W or W/O (oil/water or water/oil) emulsions inside the pump, due to the shear rate imparted and the presence of high molecular weight substances that act as natural surfactants. Therefore, it is important to perform an analysis of the flow patterns inside the pump to increase the percentage of oil recovered using the centrifugal force and the difference in density between the oil and the water to generate the separation of liquid phases. For this study, a Computational Fluid Dynamic (CFD) model was developed on STAR-CCM+ software based on 3D geometry of a Franklin Electric 4400 4' four-stage ESP. In this case, the modification of the last stage was carried out to improve the centrifugal effect inside the pump, and a perforated double tube was designed with three different holes configurations disposed at the outlet section, through which the cut water flows. The arrangement of holes used has different geometrical configurations such as circles, rectangles, and irregular shapes determined as grating around the tube. The two-phase flow was modeled using an Eulerian approach with the Volume of Fluid (VOF) method, which predicts the distribution and movement of larger interfaces in immiscible phases. Different water-oil compositions were evaluated, such as 70-30% v/v, 80-20% v/v and 90-10% v/v, respectively. Finally, greater recovery of oil was obtained. For the several compositions evaluated, the volumetric oil fraction was greater than 0.55 at the pump outlet. Similarly, it is possible to show an inversely proportional relationship between the Water/Oil rate (WOR) and the volumetric flow. The volumetric fractions evaluated, the oil flow increased approximately between 41%-10% for circular perforations and 49%-19% for rectangular shaped perforations, regarding the inlet flow. Besides, the elimination of the pump diffuser in the last stage of the pump reduced the head by approximately 20%.

Keywords: computational fluid dynamic, CFD, electrical submersible pump, ESP, two phase flow, volume of fluid, VOF, water/oil rate, WOR

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592 Planning the Journey of Unifying Medical Record Numbers in Five Facilities and the Expected Challenges: Case Study in Saudi Arabia

Authors: N. Al Khashan, H. Al Shammari, W. Al Bahli

Abstract:

Patients who are eligible to receive treatment at the National Guard Health Affairs (NGHA), Saudi Arabia will typically have four medical record numbers (MRN), one in each of the geographical areas. More hospitals and primary healthcare facilities in other geographical areas will launch soon which means more MRNs. When patients own four MRNs, this will cause major drawbacks in patients’ quality of care such as creating new medical files in different regions for relocated patients and using referral system among regions. Consequently, the access to a patient’s medical record from other regions and the interoperability of health information between the four hospitals’ information system would be challenging. Thus, there is a need to unify medical records among these five facilities. As part of the effort to increase the quality of care, a new Hospital Information Systems (HIS) was implemented in all NGHA facilities by the end of 2016. NGHA’s plan is put to be aligned with the Saudi Arabian national transformation program 2020; whereby 70% citizens and residents of Saudi Arabia would have a unified medical record number that enables transactions between multiple Electronic Medical Records (EMRs) vendors. The aim of the study is to explore the plan, the challenges and barriers of unifying the 4 MRNs into one Enterprise Patient Identifier (EPI) in NGHA hospitals by December 2018. A descriptive study methodology was used. A journey map and a project plan are created to be followed by the project team to ensure a smooth implementation of the EPI. It includes the following: 1) Approved project charter, 2) Project management plan, 3) Change management plan, 4) Project milestone dates. Currently, the HIS is using the regional MRN. Therefore, the HIS and all integrated health care systems in all regions will need modification to move from MRN to EPI without interfering with patient care. For now, the NGHA have successfully implemented an EPI connected with the 4 MRNs that work in the back end in the systems’ database.

Keywords: consumer health, health informatics, hospital information system, universal medical record number

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591 Effect of Lithium Bromide Concentration on the Structure and Performance of Polyvinylidene Fluoride (PVDF) Membrane for Wastewater Treatment

Authors: Poojan Kothari, Yash Madhani, Chayan Jani, Bharti Saini

Abstract:

The requirements for quality drinking and industrial water are increasing and water resources are depleting. Moreover large amount of wastewater is being generated and dumped into water bodies without treatment. These have made improvement in water treatment efficiency and its reuse, an important agenda. Membrane technology for wastewater treatment is an advanced process and has become increasingly popular in past few decades. There are many traditional methods for tertiary treatment such as chemical coagulation, adsorption, etc. However recent developments in membrane technology field have led to manufacturing of better quality membranes at reduced costs. This along with the high costs of conventional treatment processes, high separation efficiency and relative simplicity of the membrane treatment process has made it an economically viable option for municipal and industrial purposes. Ultrafiltration polymeric membranes can be used for wastewater treatment and drinking water applications. The proposed work focuses on preparation of one such UF membrane - Polyvinylidene fluoride (PVDF) doped with LiBr for wastewater treatment. Majorly all polymeric membranes are hydrophobic in nature. This property leads to repulsion of water and hence solute particles occupy the pores, decreasing the lifetime of a membrane. Thus modification of membrane through addition of small amount of salt such as LiBr helped us attain certain characteristics of membrane, which can then be used for wastewater treatment. The membrane characteristics are investigated through measuring its various properties such as porosity, contact angle and wettability to find out the hydrophilic nature of the membrane and morphology (surface as well as structure). Pure water flux, solute rejection and permeability of membrane is determined by permeation experiments. A study of membrane characteristics with various concentration of LiBr helped us to compare its effectivity.

Keywords: Lithium bromide (LiBr), morphology, permeability, Polyvinylidene fluoride (PVDF), solute rejection, wastewater treatment

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590 Detection and Expression of Peroxidase Genes in Trichoderma harzianum KY488466 and Its Response to Crude Oil Degradation

Authors: Michael Dare Asemoloye, Segun Gbolagade Jonathan, Rafiq Ahmad, Odunayo Joseph Olawuyi, D. O. Adejoye

Abstract:

Fungi have potentials for degrading hydrocarbons through the secretion of different enzymes. Crude oil tolerance and degradation by Trichoderma harzianum was investigated in this study with its ability to produce peroxidase enzymes (LiP and MnP). Many fungal strains were isolated from rhizosphere of grasses growing on a crude oil spilled site, and the most frequent strain based on percentage incidence was further characterized using morphological and molecular characteristics. Molecular characterization was done through the amplification of Ribosomal-RNA regions of 18s (1609-1627) and 28s (287-266) using ITS1 and ITS4 combinations and it was identified using NCBI BLAST tool. The selected fungus was also subjected to an in-vitro tolerance test at crude oil concentrations of 5, 10, 15, 20 and 25% while 0% served as control. In addition, lignin peroxidase genes (lig1-6) and manganese peroxidase gene (mnp) were detected and expressed in this strain using RT-PCR technique, its peroxidase producing activities was also studied in aliquots (U/ml). This strain had highest incidence of 80%, it was registered in NCBI as Trichoderma harzianum asemoJ KY488466. The strain KY488466 responded to crude oil concentrations as it increase, the dose inhibition response percentage (DIRP) increased from 41.67 to 95.41 at 5 to 25 % crude oil concentrations. All the peroxidase genes are present in KY488466, and expressed with amplified 900-1000 bp through RT-PCR technique. In this strain, lig2, lig4 and mnp genes were over-expressed, lig 6 was moderately expressed, while none of the genes was under-expressed. The strain also produced 90±0.87 U/ml lignin peroxidase and 120±1.23 U/mil manganese peroxidase enzymes in aliquots. These results imply that KY488466 can tolerate and survive high crude oil concentration and could be exploited for bioremediation of oil-spilled soils, the produced peroxidase enzymes could also be exploited for other biotechnological experiments.

Keywords: crude oil, enzymes, expression, peroxidase genes, tolerance, Trichoderma harzianum

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589 Effect of a GABA/5-HTP Mixture on Behavioral Changes and Biomodulation in an Invertebrate Model

Authors: Kyungae Jo, Eun Young Kim, Byungsoo Shin, Kwang Soon Shin, Hyung Joo Suh

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Gamma-aminobutyric acid (GABA) and 5-hydroxytryptophan (5-HTP) are amino acids of digested nutrients or food ingredients and these can possibly be utilized as non-pharmacologic treatment for sleep disorder. We previously investigated the GABA/5-HTP mixture is the principal concept of sleep-promoting and activity-repressing management in nervous system of D. melanogaster. Two experiments in this study were designed to evaluate sleep-promoting effect of GABA/5-HTP mixture, to clarify the possible ratio of sleep-promoting action in the Drosophila invertebrate model system. Behavioral assays were applied to investigate distance traveled, velocity, movement, mobility, turn angle, angular velocity and meander of two amino acids and GABA/5-HTP mixture with caffeine treated flies. In addition, differentially expressed gene (DEG) analyses from next generation sequencing (NGS) were applied to investigate the signaling pathway and functional interaction network of GABA/5-HTP mixture administration. GABA/5-HTP mixture resulted in significant differences between groups related to behavior (p < 0.01) and significantly induced locomotor activity in the awake model (p < 0.05). As a result of the sequencing, the molecular function of various genes has relationship with motor activity and biological regulation. These results showed that GABA/5-HTP mixture administration significantly involved the inhibition of motor behavior. In this regard, we successfully demonstrated that using a GABA/5-HTP mixture modulates locomotor activity to a greater extent than single administration of each amino acid, and that this modulation occurs via the neuronal system, neurotransmitter release cycle and transmission across chemical synapses.

Keywords: sleep, γ-aminobutyric acid, 5-hydroxytryptophan, Drosophila melanogaster

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588 Synthesis and Characterization of Capric-Stearic Acid/ Graphene Oxide-TiO₂ Microcapsules for Solar Energy Storage and Photocatalytic Efficiency

Authors: Ghada Ben Hamad, Zohir Younsi, Hassane Naji, Noureddine Lebaz, Naoual Belouaggadia

Abstract:

This study deals with a bifunctional micro-encapsulated phase change (MCP) material, capric-stearic acid/graphene oxide-TiO2, which has been successfully developed by in situ hydrolysis and polycondensation of tetrabutyl titanate and modification of graphene oxide (GO) on the TiO2 doped shell. The use of graphene and doped TiO2 is a promising approach to provide photocatalytic activity under visible light and improve the microcapsules physicochemical properties. The morphology and chemical structure of the resulting microcapsule samples were determined by using Fourier transform infrared (FT-IR) spectroscopy, scanning electronic microscope (SEM), and X-ray diffractometer (XRD) methods. The ultraviolet, visible spectrophotometer (UV–vis), the differential scanning calorimeter (DSC) and the thermogravimetric analyzer (TGA) were used to investigate the absorption of visible and ultraviolet (UV), the thermal properties, and thermal stabilities of the microcapsules. Note that, the visible light photocatalytic activity was assessed for the toluene and benzene gaseous removal in a suitable test room. The microcapsules exhibit an interesting spherical morphology and an average diameter of 15 to 25 μm. The addition of graphene can enhance the rigidity of the shell and improve the microcapsules thermal reliability. At the same time, the thermal analysis tests showed that the synthesized microcapsules had a high solar thermal energy-storage and better thermal stability. In addition, the capric-stearic acid microcapsules exhibited high solar photocatalytic activity with respect to atmospheric pollutants under natural sunlight. The fatty acid samples obtained with the GO/TiO2 shell showed great potential for applications of solar energy storage, solar photocatalytic degradation of air pollutants and buildings energy conservation.

Keywords: thermal energy storage, microencapsulation, titanium dioxide, photocatalysis, graphene oxide

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587 Stem Cell Differentiation Toward Secretory Progenitors after Intestinal Ischemia-Reperfusion in a Rat is Accompanied by Inhibited Notch Signaling Cascade

Authors: Igor Sukhotnik

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Objectives: Notch signaling is thought to act to drive cell versification in the lining of the small intestine. When Notch signaling is blocked, proliferation ceases, and epithelial cells become secretory. The purpose of the present study was to evaluate the role of Notch signaling pathway in stem cell differentiation in a rat model of intestinal ischemia-reperfusion (IR). Methods: Male Sprague-Dawley rats were randomly divided into four experimental groups: Sham-24 and Sham-48 rats underwent laparotomy and were killed 24 or 48 h later, respectively; IR-24 and IR-48 rats underwent occlusion of SMA and portal vein for 30 min followed by 24 or 48 h of reperfusion, respectively. Notch-related gene and protein expression were determined using Real Time PCR, Western blotting and immunohistochemistry. Wax histology and immunohistochemistry was used to determine cell differentiation toward absorptive (enterocytes) or secretory progenitors (goblet cells, enteroendocrine cells or Paneth cells). Results: IR-48 rats exhibited a significant decrease in Notch-1 protein expression (Western blot) that was coincided with a significant decrease in the number of Notch-1 positive cells (immunohistochemistry) in jejunum and ileum as well as Hes-1 positive cells in jejunum and ileum compared to Sham-48 rats. A significant down-regulation of Notch signaling related genes and proteins in IR animals was accompanied by a significant increase in the number of goblet and Paneth cells and decreased number of absorptive cells compared to control rats. Conclusions: Forty-eight hours following intestinal IR in rats, inhibited Notch signaling pathway was accompanied by intestinal stem cells differentiation toward secretory progenitors.

Keywords: Intestine, notch, ischemia-reperfusion, cell differentiation, secretory

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586 A New Binder Mineral for Cement Stabilized Road Pavements Soils

Authors: Aydın Kavak, Özkan Coruk, Adnan Aydıner

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Long-term performance of pavement structures is significantly impacted by the stability of the underlying soils. In situ subgrades often do not provide enough support required to achieve acceptable performance under traffic loading and environmental demands. NovoCrete® is a powder binder-mineral for cement stabilized road pavements soils. NovoCrete® combined with Portland cement at optimum water content increases the crystallize formations during the hydration process, resulting in higher strengths, neutralizes pH levels, and provides water impermeability. These changes in soil properties may lead to transforming existing unsuitable in-situ materials into suitable fill materials. The main features of NovoCrete® are: They are applicable to all types of soil, reduce premature cracking and improve soil properties, creating base and subbase course layers with high bearing capacity by reducing hazardous materials. It can be used also for stabilization of recyclable aggregates and old asphalt pavement aggregate, etc. There are many applications in Germany, Turkey, India etc. In this paper, a few field application in Turkey will be discussed. In the road construction works, this binder material is used for cement stabilization works. In the applications 120-180 kg cement is used for 1 m3 of soil with a 2 % of binder NovoCrete® material for the stabilization. The results of a plate loading test in a road construction site show 1 mm deformation which is very small under 7 kg/cm2 loading. The modulus of subgrade reaction increase from 611 MN/m3 to 3673 MN/m3.The soaked CBR values for stabilized soils increase from 10-20 % to 150-200 %. According to these data weak subgrade soil can be used as a base or sub base after the modification. The potential reduction in the need for quarried materials will help conserve natural resources. The use of on-site or nearby materials in fills, will significantly reduce transportation costs and provide both economic and environmental benefits.

Keywords: soil, stabilization, cement, binder, Novocrete, additive

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585 Investigating the Effect of Ceramic Thermal Barrier Coating on Diesel Engine with Lemon Oil Biofuel

Authors: V. Karthickeyan

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The demand for energy is anticipated to increase, due to growing urbanization, industrialization, upgraded living standards and cumulatively increasing human population. The general public is becoming gradually aware of the diminishing fossil fuel resources along with the environmental issues, and it has become clear that biofuel is intended to make significant support to the forthcoming energy needs of the native and industrial sectors. Nowadays, the investigation on biofuels obtained from peels of fruits and vegetables have gained the consideration as an environment-friendly alternative to diesel. In the present work, biofuel was produced from non-edible Lemon Oil (LO) using steam distillation process. LO is characterized by its beneficial aspects like low kinematic viscosity and enhanced calorific value which provides better fuel atomization and evaporation. Furthermore, the heating values of the biofuels are approximately equal to diesel. A single cylinder, four-stroke diesel engine was used for this experimentation. An engine modification technique namely Thermal Barrier Coating (TBC) was attempted. Combustion chamber components were thermally coated with ceramic material namely partially stabilized zirconia (PSZ). The benefit of thermal barrier coating is to diminish the heat loss from engine and transform the collected heat into piston work. Performance characteristics like Brake Thermal Efficiency (BTE) and Brake Specific Fuel Consumption (BSFC) were analyzed. Combustion characteristics like in-cylinder pressure and heat release rate were analyzed. In addition, the following engine emissions namely nitrogen oxide (NO), carbon monoxide (CO), hydrocarbon (HC), and smoke were measured. The acquired performance combustion and emission characteristics of uncoated engine were compared with PSZ coated engine. From the results, it was perceived that the LO biofuel may be considered as the prominent alternative in the near prospect with thermal barrier coating technique to enrich the performance, combustion and emission characteristics of diesel engine.

Keywords: ceramic material, thermal barrier coating, biofuel and diesel engine

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584 Biophysical Study of the Interaction of Harmalol with Nucleic Acids of Different Motifs: Spectroscopic and Calorimetric Approaches

Authors: Kakali Bhadra

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Binding of small molecules to DNA and recently to RNA, continues to attract considerable attention for developing effective therapeutic agents for control of gene expression. This work focuses towards understanding interaction of harmalol, a dihydro beta-carboline alkaloid, with different nucleic acid motifs viz. double stranded CT DNA, single stranded A-form poly(A), double-stranded A-form of poly(C)·poly(G) and clover leaf tRNAphe by different spectroscopic, calorimetric and molecular modeling techniques. Results of this study converge to suggest that (i) binding constant varied in the order of CT DNA > poly(C)·poly(G) > tRNAphe > poly(A), (ii) non-cooperative binding of harmalol to poly(C)·poly(G) and poly(A) and cooperative binding with CT DNA and tRNAphe, (iii) significant structural changes of CT DNA, poly(C)·poly(G) and tRNAphe with concomitant induction of optical activity in the bound achiral alkaloid molecules, while with poly(A) no intrinsic CD perturbation was observed, (iv) the binding was predominantly exothermic, enthalpy driven, entropy favoured with CT DNA and poly(C)·poly(G) while it was entropy driven with tRNAphe and poly(A), (v) a hydrophobic contribution and comparatively large role of non-polyelectrolytic forces to Gibbs energy changes with CT DNA, poly(C)·poly(G) and tRNAphe, and (vi) intercalated state of harmalol with CT DNA and poly(C)·poly(G) structure as revealed from molecular docking and supported by the viscometric data. Furthermore, with competition dialysis assay it was shown that harmalol prefers hetero GC sequences. All these findings unequivocally pointed out that harmalol prefers binding with ds CT DNA followed by ds poly(C)·poly(G), clover leaf tRNAphe and least with ss poly(A). The results highlight the importance of structural elements in these natural beta-carboline alkaloids in stabilizing different DNA and RNA of various motifs for developing nucleic acid based better therapeutic agents.

Keywords: calorimetry, docking, DNA/RNA-alkaloid interaction, harmalol, spectroscopy

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583 Entry Inhibitors Are Less Effective at Preventing Cell-Associated HIV-2 Infection than HIV-1

Authors: A. R. Diniz, P. Borrego, I. Bártolo, N. Taveira

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Cell-to-cell transmission plays a critical role in the spread of HIV-1 infection in vitro and in vivo. Inhibition of HIV-1 cell-associated infection by antiretroviral drugs and neutralizing antibodies (NAbs) is more difficult compared to cell-free infection. Limited data exists on cell-associated infection by HIV-2 and its inhibition. In this work, we determined the ability of entry inhibitors to inhibit HIV-1 and HIV-2 cell-to cell fusion as a proxy to cell-associated infection. We developed a method in which Hela-CD4-cells are first transfected with a Tat expressing plasmid (pcDNA3.1+/Tat101) and infected with recombinant vaccinia viruses expressing either the HIV-1 (vPE16: from isolate HTLV-IIIB, clone BH8, X4 tropism) or HIV-2 (vSC50: from HIV-2SBL/ISY, R5 and X4 tropism) envelope glycoproteins (M.O.I.=1 PFU/cell).These cells are added to TZM-bl cells. When cell-to-cell fusion (syncytia) occurs the Tat protein diffuses to the TZM-bl cells activating the expression of a reporter gene (luciferase). We tested several entry inhibitors including the fusion inhibitors T1249, T20 and P3, the CCR5 antagonists MVC and TAK-779, the CXCR4 antagonist AMD3100 and several HIV-2 neutralizing antibodies (Nabs). All compounds inhibited HIV-1 and HIV-2 cell fusion albeit to different levels. Maximum percentage of HIV-2 inhibition (MPI) was higher for fusion inhibitors (T1249- 99.8%; P3- 95%, T20-90%) followed by co-receptor antagonists (MVC- 63%; TAK-779- 55%; AMD3100- 45%). NAbs from HIV-2 infected patients did not prevent cell fusion up to the tested concentration of 4μg/ml. As for HIV-1, MPI reached 100% with TAK-779 and T1249. For the other antivirals, MPIs were: P3-79%; T20-75%; AMD3100-61%; MVC-65%.These results are consistent with published data. Maraviroc had the lowest IC50 both for HIV-2 and HIV-1 (IC50 HIV-2= 0.06 μM; HIV-1=0.0076μM). Highest IC50 were observed with T20 for HIV-2 (3.86μM) and with TAK-779 for HIV-1 (12.64μM). Overall, our results show that entry inhibitors in clinical use are less effective at preventing Env mediated cell-to-cell-fusion in HIV-2 than in HIV-1 which suggests that cell-associated HIV-2 infection will be more difficult to inhibit compared to HIV-1. The method described here will be useful to screen for new HIV entry inhibitors.

Keywords: cell-to-cell fusion, entry inhibitors, HIV, NAbs, vaccinia virus

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582 In situ Ortho-Quinone Methide Reactions for Construction of Flavonoids with Fused Ring Systems

Authors: Vidia A. Nuraini, Eugene M. H. Yee, Mohan Bhadbhade, David StC. Black, Naresh Kumar

Abstract:

Flavonoids are naturally occurring compounds that have been shown to exhibit a wide range of biological properties including anticancer and anti-inflammatory activities. However, flavonoids suffer from low bioavailability, which limits their overall utility for therapeutic applications. One of the methods to overcome this limitation is through structural modification of natural flavonoids. In this study, flavanone, isoflavanone, and isoflavene, were structurally modified through the introduction of additional fused-ring systems via ortho-quinone methide intermediates (o-QMs). These intermediates can readily undergo a [4+2] cycloaddition through an inverse-electron-demand Diels–Alder reaction with electron-rich dienophiles. A regioselective Mannich reaction using bis-(N,N-dimethylamino)methane was employed to generate the o-QM precursors of flavanone, isoflavanone, and isoflavene. The o-QM intermediates were subsequently generated in situ through thermal elimination of the dimethylamine functionality and reacted with a variety of dienophiles to produce novel flavonoids with fused-ring systems. A total of 21 novel flavonoid analogs were successfully synthesized. The X-ray crystal structure of cycloaddition adducts, particularly those derived from 3,4-dihydro-2H-pyran and p-methoxystyrene revealed a special case of enantiomeric disorder, where two enantiomers in equal amounts superpose with one another, with the exception for atoms that have opposite configuration. The anticancer properties of fused-ring systems derived from isoflavene were evaluated against the neuroblastoma SKN-BE(2)C, the triple negative breast cancer MDA-MB-231, and the glioblastoma U87 cancer cell lines. One of these cycloaddition adducts had displayed improved anti-proliferative activity against MDA-MB-231 and U87 cancer cell lines as compared to the parent compound. Further anticancer and anti-inflammatory activities of the flavanone and isoflavanone analogs are currently being investigated.

Keywords: Diels-Alder reaction, flavonoids, Mannich reaction, ortho-quinone methide.

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581 Using Baculovirus Expression Vector System to Express Envelop Proteins of Chikungunya Virus in Insect Cells and Mammalian Cells

Authors: Tania Tzong, Chao-Yi Teng, Tzong-Yuan Wu

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Currently, Chikungunya virus (CHIKV) transmitted to humans by Aedes mosquitoes has distributed from Africa to Southeast Asia, South America, and South Europe. However, little is known about the antigenic targets for immunity, and there are no licensed vaccines or specific antiviral treatments for the disease caused by CHIKV. Baculovirus has been recognized as a novel vaccine vector with attractive characteristic features of an optional vaccine delivery vehicle. This approach provides the safety and efficacy of CHIKV vaccine. In this study, bi-cistronic recombinant baculoviruses vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP were produced. Both recombinant baculovirus can express EGFP reporter gene in insect cells to facilitate the recombinant virus isolation and purification. Examination of vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP showed that this recombinant baculovirus could induce syncytium formation in insect cells. Unexpectedly, the immunofluorescence assay revealed the expression of E1 and E2 of CHIKV structural proteins in insect cells infected by vAc-CMV-CHIKV26S-Rhir-EGFP. This result may imply that the CMV promoter can induce the transcription of CHIKV26S in insect cells. There are also E1 and E2 expression in mammalian cells transduced by vAc-CMV-CHIKV26S-Rhir-EGFP and vAc-CMV-pH-CHIKV26S-Lir-EGFP. The expression of E1 and E2 proteins of insect and mammalian cells was validated again by Western blot analysis. The vector construction with dual tandem promoters, which is polyhedrin and CMV promoter, has higher expression of the E1 and E2 of CHIKV structural proteins than the vector construction with CMV promoter only. Most of the E1 and E2 proteins expressed in mammalian cells were glycosylated. In the future, the expression of structural proteins of CHIKV in mammalian cells is expected can form virus-like particle, so it could be used as a vaccine for chikungunya virus.

Keywords: chikungunya virus, virus-like particle, vaccines, baculovirus expression vector system

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580 Deficiencies in Vitamin A and Iron Supply Potential of Selected Indigenous Complementary Foods of Infants in Uganda

Authors: Richard Kajjura, Joyce Kikafunda, Roger Whitehead

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Introduction: Indigenous complementary recipes for children (6-23 months) are bulky and inextricably linked. The potential contribution of indigenous complementary foods to infant’s vitamin A and iron needs is not well investigated in Uganda. Less is known whether children in Uganda are living with or without adequate supply of vitamin A and iron nutrients. In this study, vitamin A and iron contents were assessed in the complementary foods fed to infants aged 6-11 months in a Peri-urban setting in Kampala District in Central Uganda. Objective: Assessment of vitamin A and iron contents of indigenous complementary foods of children as fed and associated demographic factor. Method: In a cross sectional study design, one hundred and three (153) households with children aged 6-11 months were randomly selected to participate in the assessment. Complementary food samples were collected from the children’s mothers/caretakers at the time of feeding the child. The mothers’ socio-demographic characteristics of age, education, marital status, occupation and sex collected a semi-qualitative questionnaire. The Vitamin A and iron contents in the complementary foods were analyzed using a UV/VIS spectrophotometer for vitamin A and Atomic Absorption spectrophotometer for iron samples. The data was analyzed using Gene-stat software program. Results: The mean vitamin A content was 97.0± 72.5 µg while that of iron was 1.5 ± 0.4 mg per 100g of food sample as fed. The contribution of indigenous complementary foods found was 32% for vitamin A and 15% iron of the recommended dietary allowance. Age of children was found to be significantly associated Vitamin A and Iron supply potential. Conclusion: The contribution of indigenous complementary foods to infant’s vitamin A and iron needs was low. Complementary foods in Uganda are more likely to be deficient in vitamin A and iron content. Nutrient dense dietary supplementation should be intervened in to make possible for Ugandan children attain full growth potential.

Keywords: indigenous complementary food, infant, iron, vitamin A

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579 Elevated Creatinine Clearance and Normal Glomerular Filtration Rate in Patients with Systemic Lupus erythematosus

Authors: Stoyanka Vladeva, Elena Kirilova, Nikola Kirilov

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Background: The creatinine clearance is a widely used value to estimate the GFR. Increased creatinine clearance is often called hyperfiltration and is usually seen during pregnancy, patients with diabetes mellitus preceding the diabetic nephropathy. It may also occur with large dietary protein intake or with plasma volume expansion. Renal injury in lupus nephritis is known to affect the glomerular, tubulointerstitial, and vascular compartment. However high creatinine clearance has not been found in patients with SLE, Target: Follow-up of creatinine clearance values in patients with systemic lupus erythematosus without history of kidney injury. Material and methods: We observed the creatinine, creatinine clearance, GFR and dipstick protein values of 7 women (with a mean age of 42.71 years) with systemic lupus erythematosus. Patients with active lupus have been monthly tested in the period of 13 months. Creatinine clearance has been estimated by Cockcroft-Gault Equation formula in ml/sec. GFR has been estimated by MDRD formula (The Modification of Diet in renal Disease) in ml/min/1.73 m2. Proteinuria has been defined as present when dipstick protein > 1+.Results: In all patients without history of kidney injury we found elevated creatinine clearance levels, but GFRremained within the reference range. Two of the patients were in remission while the other five patients had clinically and immunologically active Lupus. Three of the patients had a permanent presence of high creatinine clearance levels and proteinuria. Two of the patients had periodically elevated creatinine clearance without proteinuria. These results show that kidney disturbances may be caused by the vascular changes typical for SLE. Glomerular hyperfiltration can be result of focal segmental glomerulosclerosis caused by a reduction in renal mass. Probably lupus nephropathy is preceded not only by glomerular vascular changes, but also by tubular vascular changes. Using only the GFR is not a sufficient method to detect these primary functional disturbances. Conclusion: For early detection of kidney injury in patients with SLE we determined that the follow up of creatinine clearance values could be helpful.

Keywords: systemic Lupus erythematosus, kidney injury, elevated creatinine clearance level, normal glomerular filtration rate

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578 An Unbiased Profiling of Immune Repertoire via Sequencing and Analyzing T-Cell Receptor Genes

Authors: Yi-Lin Chen, Sheng-Jou Hung, Tsunglin Liu

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Adaptive immune system recognizes a wide range of antigens via expressing a large number of structurally distinct T cell and B cell receptor genes. The distinct receptor genes arise from complex rearrangements called V(D)J recombination, and constitute the immune repertoire. A common method of profiling immune repertoire is via amplifying recombined receptor genes using multiple primers and high-throughput sequencing. This multiplex-PCR approach is efficient; however, the resulting repertoire can be distorted because of primer bias. To eliminate primer bias, 5’ RACE is an alternative amplification approach. However, the application of RACE approach is limited by its low efficiency (i.e., the majority of data are non-regular receptor sequences, e.g., containing intronic segments) and lack of the convenient tool for analysis. We propose a computational tool that can correctly identify non-regular receptor sequences in RACE data via aligning receptor sequences against the whole gene instead of only the exon regions as done in all other tools. Using our tool, the remaining regular data allow for an accurate profiling of immune repertoire. In addition, a RACE approach is improved to yield a higher fraction of regular T-cell receptor sequences. Finally, we quantify the degree of primer bias of a multiplex-PCR approach via comparing it to the RACE approach. The results reveal significant differences in frequency of VJ combination by the two approaches. Together, we provide a new experimental and computation pipeline for an unbiased profiling of immune repertoire. As immune repertoire profiling has many applications, e.g., tracing bacterial and viral infection, detection of T cell lymphoma and minimal residual disease, monitoring cancer immunotherapy, etc., our work should benefit scientists who are interested in the applications.

Keywords: immune repertoire, T-cell receptor, 5' RACE, high-throughput sequencing, sequence alignment

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577 Analysis of Extracellular Vesicles Interactomes of two Isoforms of Tau Protein via SHSY-5Y Cell Lines

Authors: Mohammad Aladwan

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Alzheimer’s disease (AD) is a widespread dementing illness with a complex and poorly understood etiology. An important role in improving our understanding of the AD process is the modeling of disease-associated changes in tau protein phosphorylation, a protein known to mediate events essential to the onset and progression of AD. A main feature of AD is the abnormal phosphorylation of tau protein and the presence of neurofibrillary tangles. In order to evaluate the respective roles of the microtubule-binding region (MTBR) and alternatively spliced exons in the N-terminal projection domains in AD, we have constructed SHSY-5Y cell lines that stably overexpress four different species of tau protein (4R2N, 4R0N, N(E-2), N(E+2)). Since the toxicity and spreading of tau lesions in AD depends on the interactions of tau with other proteins, we have performed a proteomic analysis of exosome-fraction interactomes for cell lysates and media samples that were isolated from SHSY-5Y cell lines. Functional analysis of tau interactomes based on gene ontology (GO) terms was performed using the String 10.5 database program. The highest number of exosomes proteomes and tau associated proteins were found with 4R2N isoform (2771 and 159) in cell lysate and they have a high strength of connectivity (78%) between proteins, while N(E-2) isoform in the media proteomes has the highest number of proteins and tau associated protein (1829 and 205). Moreover, known AD markers were significantly enriched in secreted interactomes relative to lysate interactomes in the SHSY-5Y cells of tau isoforms lacking exons 2 and 3 in the N-terminal. The lack of exon 2 (E-2) from tau protein can be mediated by tau secretion and spreading to different cells. Enriched functions in the secreted E-2 interactome include signaling and developmental pathways that have been linked to a) tau misprocessing and lesion development and b) tau secretion and which, therefore, could play novel roles in AD pathogenesis.

Keywords: Alzheimer's disease, dementia, tau protein, neurodegenration disease

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576 The Psychosis Prodrome: Biomarkers of the Glutamatergic System and Their Potential Role in Prediction and Treatment

Authors: Peter David Reiss

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The concept of the psychosis prodrome has allowed for the identification of adolescent and young adult patients who have a significantly elevated risk of developing schizophrenia spectrum disorders. A number of different interventions have been tested in order to prevent or delay progression of symptoms. To date, there has been no consistent meta-analytical evidence to support efficacy of antipsychotic treatment for patients in the prodromal state, and their use remains therefore inconclusive. Although antipsychotics may manage symptoms transiently, they have not been found to prevent or delay onset of psychotic disorders. Furthermore, pharmacological intervention in high-risk individuals remains controversial, because of the antipsychotic side effect profile in a population in which only about 20 to 35 percent will eventually convert to psychosis over a two-year period, with even after two years conversion rates not exceeding 30 to 40 percent. This general estimate is additionally problematic, in that it ignores the fact that there is significant variation in individual risk among clinical high-risk cases. The current lack of reliable tests for at-risk patients makes it difficult to justify individual treatment decisions. Preventive treatment should ideally be dictated by an individual’s risk while minimizing potentially harmful medication exposure. This requires more accurate predictive assessments by using valid and accessible prognostic markers. The following will compare prediction and risk modification potential of behavioral biomarkers such as disturbances of basic sense of self and emotion awareness, neurocognitive biomarkers such as attention, working and declarative memory, and neurophysiological biomarkers such as glutamatergic abnormalities and NMDA receptor dysfunction. Identification of robust biomarkers could therefore not only provide more reliable means of psychosis prediction, but also help test and develop new clinical interventions targeted at the prodromal state.

Keywords: at-risk mental state, biomarkers, glutamatergic system, NMDA receptor, psychosis prodrome, schizophrenia

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575 Synergistic Effect of Curcumin and Insulin on GLUT4 Translocation in C2C12 Cell

Authors: Javad Mohiti-Ardekani, Shabodin Asadii, Ali Moradi

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Introduction: Curcumin, the yellow pigment in turmeric, has been shown as an anti-diabetic agent for centuries but only in recent few years, its mechanism of action has been under investigation. Some studies showed that curcumin might exert its anti-diabetic effect via increasing glucose transporter isotype-4 (GLUT4) gene and glycoprotein contents in cells. To investigate this possibility, we investigate the effects of extract and commercial curcumin with and without insulin on GLUT4 translocation from intracellular compartments of nuclear or endoplasmic reticulum membranes (N/ER) into the cytoplasmic membrane (CM). Methods and Material: C2C12 myoblastic cell line were seeded in DMEM plus 20 % FBS and differentiated to myotubes using 2 % horse serum. After myotubes formation, 40 µmolar Extract and Commercial curcumin, with or without insulin as intervention, and as control 1 % DMSO were added for 3 h. Cells were washed and homogenized followed by ultracentrifuge fractionation, protein separation by SDS-PAGE and GLUT4 detection using semi-quantitative Western blotting. Data analysis was done by two independent samples t-test for comparison of mean ± SD of GLUT4 percent in categories. GLUT4 contents were higher in CM groups curcumin and curcumin with insulin in comparison to 1 % DMSO-treated myotubes control group. Results: As our results have shown extract and commercial curcumin induces GLUT4 translocation from intra-cell into cell surface. The results have also shown synergic effect of curcumin on translocation of GLUT4 from intra-cell into cell surface in the presence of 100 nm insulin. Discussion: We conclude that curcumin may be a choice of type-2 diabetes mellitus treatment because its extract and commercial enhances GLUT4 contents in CM where it facilitates glucose entrance into the cell. However, it is necessary to trace the signaling pathways which are activated by curcumin.

Keywords: Curcumin, insulin, Diabetes type-2, GLUT4

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574 Endophytic Fungi Recovered from Lycium arabicum as an Eco-Friendly Alternative for Fusarium Crown and Root Rot Disease Control and Tomato Growth Enhancement

Authors: Ahlem Nefzi, Rania Aydi Ben Abdallah, Hayfa Jabnoun-Khiareddine, Ammar Nawaim, Rabiaa Haouala, Mejda Daami-Remadi

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Seven endophytic fungi were isolated from the wild Solanaceous species Lycium arabicum growing in the Tunisian Centre-East and were assessed for their ability to suppress Fusarium Crown and Root Rot disease caused by Fusarium oxysporum f. sp. radicis lycopersici (FORL) and to enhance plant growth. Fungal isolates were shown able to colonize tomato cv. Rio Grande roots, crowns, and stems. A significant promotion in all studied growth parameters (root length, shoot height, and roots and shoots fresh weight) was recorded in tomato plants treated with fungal conidial suspensions or their cell-free culture filtrates compared to FORL-inoculated or pathogen-free controls. I15 and I18 isolates were shown to be the most effective leading to 85.7-87.5 and 93.6-98.4% decrease in leaf and root damage index and the vascular discoloration extent, respectively, over FORL-inoculated and untreated control. These two bioactive and growth-promoting isolates (I15 and I18) were morphologically characterized and identified using rDNA sequencing gene as being Alternaria alternata (MF693801) and Fusarium fujikuroi (MF693802). These fungi significantly suppressed FORL mycelial growth and showed chitinolytic, proteolytic and amylase activities whereas only F. fujikuroi displayed a lipolytic activity. This study clearly demonstrated the potential use of fungi naturally associated with L. arabicum as biocontrol and bio-fertilizing agents.

Keywords: biocontrol, endophytic fungi, Fusarium oxysporum f. sp. radicis-lycopersici, tomato promotion, Lycium arabicum

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573 Tripeptide Inhibitor: The Simplest Aminogenic PEGylated Drug against Amyloid Beta Peptide Fibrillation

Authors: Sutapa Som Chaudhury, Chitrangada Das Mukhopadhyay

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Alzheimer’s disease is a well-known form of dementia since its discovery in 1906. Current Food and Drug Administration approved medications e.g. cholinesterase inhibitors, memantine offer modest symptomatic relief but do not play any role in disease modification or recovery. In last three decades many small molecules, chaperons, synthetic peptides, partial β-secretase enzyme blocker have been tested for the development of a drug against Alzheimer though did not pass the 3rd clinical phase trials. Here in this study, we designed a PEGylated, aminogenic, tripeptidic polymer with two different molecular weights based on the aggregation prone amino acid sequence 17-20 in amyloid beta (Aβ) 1-42. Being conjugated with poly-ethylene glycol (PEG) which self-assembles into hydrophilic nanoparticles, these PEGylated tripeptides constitute a very good drug delivery system crossing the blood brain barrier while the peptide remains protected from proteolytic degradation and non-specific protein interactions. Moreover, being completely aminogenic they would not raise any side effects. These peptide inhibitors were evaluated for their effectiveness against Aβ42 fibrillation at an early stage of oligomer to fibril formation as well as preformed fibril clearance via Thioflavin T (ThT) assay, dynamic light scattering analyses, atomic force microscopy and scanning electron microscopy. The inhibitors were proved to be safe at a higher concentration of 20µM by the reduction assay of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) dye. Moreover, SHSY5Y neuroblastoma cells have shown a greater survivability when treated with the inhibitors following Aβ42 fibril and oligomer treatment as compared with the control Aβ42 fibril and/or oligomer treated neuroblastoma cells. These make the peptidic inhibitors a promising compound in the aspect of the discovery of alternative medication for Alzheimer’s disease.

Keywords: Alzheimer’s disease, alternative medication, amyloid beta, PEGylated peptide

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572 Identification of the Putative Interactome of Escherichia coli Glutaredoxin 2 by Affinity Chromatography

Authors: Eleni Poulou-Sidiropoulou, Charalampos N. Bompas, Martina Samiotaki, Alexios Vlamis-Gardikas

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The glutaredoxin (Grx) and thioredoxin (Trx) systems keep the intracellular environment reduced in almost all organisms. In Escherichia coli (E. coli), the Grx system relies on NADPH+ to reduce GSH reductase (GR), the latter reducing oxidized diglutathione to glutathione (GSH) which in turn reduces cytosolic Grxs, the electron donors for different intracellular substrates. In the Trx system, GR and GSH are replaced by Trx reductase (TrxR). Three of the Grxs of E. coli (Grx1, 2, 3) are reduced by GSH, while Grx4 is likely reduced by TrxR. Trx1 and Grx1 from E. coli may reduce ribonucleotide reductase Ia to ensure a constant supply of deoxyribonucleotides for the synthesis of DNA. The role of the other three Grxs is relatively unknown, especially for Grx2 that may amount up to 1 % of total cellular protein in the stationary phase of growth. The protein is known as a potent antioxidant, but no specific functions have been attributed to it. Herein, affinity chromatography of cellular extracts on immobilized Grx2, followed by MS analysis of the resulting eluates, was employed to identify protein ligands that could provide insights into the biological role of Grx2. Ionic, strong non-covalent, and covalent (disulfide) interactions with relevant proteins were detected. As a means of verification, the identified ligands were subjected to in silico docking with monothiol Grx2. In other experiments, protein extracts from E. coli cells lacking the gene for Grx2 (grxB) were compared to those of wild type. Taken together, the two approaches suggest that Grx2 is involved in protein synthesis, nucleotide metabolism, DNA damage repair, stress responses, and various metabolic processes. Grx2 appears as a versatile protein that may participate in a wide range of biological pathways beyond its known general antioxidant function.

Keywords: Escherichia coli, glutaredoxin 2, interactome, thiol-disulfide oxidoreductase

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571 Synthesis and Characterization of the Carbon Spheres Built Up from Reduced Graphene Oxide

Authors: Takahiro Saida, Takahiro Kogiso, Takahiro Maruyama

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The ordered structural carbon (OSC) material is expected to apply to the electrode of secondary batteries, the catalyst supports, and the biomaterials because it shows the low substance-diffusion resistance by its uniform pore size. In general, the OSC material is synthesized using the template material. Changing size and shape of this template provides the pore size of OSC material according to the purpose. Depositing the oxide nanosheets on the polymer sphere template by the layer by layer (LbL) method was reported as one of the preparation methods of OSC material. The LbL method can provide the controlling thickness of structural wall without the surface modification. When the preparation of the uniform carbon sphere prepared by the LbL method which composed of the graphene oxide wall and the polymethyl-methacrylate (PMMA) core, the reduction treatment will be the important object. Since the graphene oxide has poor electron conductivity due to forming a lot of functional groups on the surface, it could be hard to apply to the electrode of secondary batteries and the catalyst support of fuel cells. In this study, the graphene oxide wall of carbon sphere was reduced by the thermal treatment under the vacuum conditions, and its crystalline structure and electronic state were characterized. Scanning electron microscope images of the carbon sphere after the heat treatment at 300ºC showed maintaining sphere shape, but its shape was collapsed with increasing the heating temperature. In this time, the dissolution rate of PMMA core and the reduction rate of graphene oxide were proportionate to heating temperature. In contrast, extending the heating time was conducive to the conservation of the sphere shape. From results of X-ray photoelectron spectroscopy analysis, its electronic state of the surface was indicated mainly sp² carbon. From the above results, we succeeded in the synthesis of the sphere structure composed by the reduction graphene oxide.

Keywords: carbon sphere, graphene oxide, reduction, layer by layer

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570 Association of MMP-2,-9 Overexpression and Imbalance PGR-A/PGR-B Ratio in Endometriosis

Authors: P. Afsharian, S. Mousazadeh, M. Shahhoseini, R. Aflatoonian

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Introduction: Matrix MetalloProteinases (MMPs) degrade extracellular matrix components to provide normal remodeling and contribute to pathological tissue destruction and cell migration in endometriosis. It is accepted that MMPs are resistant to suppression by progesterone in endometriotic tissues. The physiological effects of progesterone are mediated by its two progesterone receptor (PGR) isoforms, namely PGR-A and PGR-B. The capacity of progesterone affect to gene expression is dependent on the PGR-A/PGR-B ratio. The imbalance ratio in endometriotic tissue may be an important mechanism to be resulted in Progesterone resistance and modify progesterone action via differential regulation of specific progesterone response genes and improve endometriosis disease. Material and methods: RNA was extracted from twenty ectopic (endometriotic) and eutopic (endometrial) tissue samples of women undergoing laparoscopy for endometriosis and 20 healthy fertile women at Royan Institute, Tehran, Iran. Analysis of PGR-A, PGR-B, MMP-2 and MMP-9 mRNA expression was performed using Real-time PCR in ectopic and eutopic tissues. Then, Statistical analysis was calculated according to the 2-ΔΔCT equation for all samples. Results: Quantitative RT–PCR analyses of PGR-A and PGR-B mRNA revealed that there were differences in both isoformes of PGRs mRNA expressions between ectopic and control eutopic tissues. We were able to demonstrate low expression levels of PGR-B isoforms in ectopic tissues. Although, PGR-A expression was significantly higher in the same ectopic samples compare to controls.This method permitted us to demonstrate significant overexpression of MMP-2 and MMP-9 in ectopic samples compared to control endometrial tissues, as well. Conclusions: Our data suggest that low expression levels of PGR-B and overexpression of PGR-A can alter PGR-A/PGR-B ratio in endometriotic ectopic tissues. Imbalance ratio of PGRs in endometriotic tissue may be able to consequence MMP-2 and MMP-9 overexpression which can be important in pathogenesis and treatment of disease.

Keywords: endometriosis, matrix metalloproteinases, progesterone receptor -A and -B, PGR-A/PGR-B ratio

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569 The Predictive Value of Micro Rna 451 on the Outcome of Imatinib Treatment in Chronic Myeloid Leukemia Patients

Authors: Nehal Adel Khalil, Amel Foad Ketat, Fairouz Elsayed Mohamed Ali, Nahla Abdelmoneim Hamid, Hazem Farag Manaa

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Background: Chronic myeloid leukemia (CML) represents 15% of adult leukemias. Imatinib Mesylate (IM) is the gold standard treatment for new cases of CML. Treatment with IM results in improvement of the majority of cases. However, about 25% of cases may develop resistance. Sensitive and specific early predictors of IM resistance in CML patients have not been established to date. Aim: To investigate the value of miR-451 in CML as an early predictor for IM resistance in Egyptian CML patients. Methods: The study employed Real time Polymerase Reaction (qPCR) technique to investigate the leucocytic expression of miR-451 in fifteen newly diagnosed CML patients (group I), fifteen IM responder CML patients (group II), fifteen IM resistant CML patients (group III) and fifteen healthy subjects of matched age and sex as a control group (group IV). The response to IM was defined as < 10% BCR-ABL transcript level after 3 months of therapy. The following parameters were assessed in subjects of all the studied groups: 1- Complete blood count (CBC). 2- Measurement of plasma level of miRNA 451 using real-time Polymerase Chain Reaction (qPCR). 3- Detection of BCR-ABL gene mutation in CML using qPCR. Results: The present study revealed that miR-451 was significantly down-regulated in leucocytes of newly diagnosed CML patients as compared to healthy subjects. IM responder CML patients showed an up-regulation of miR- 451 compared with IM resistant CML patients. Conclusion: According to the data from the present study, it can be concluded that leucocytic miR- 451 expression is a useful additional follow-up marker for the response to IM and a promising prognostic biomarker for CML.

Keywords: chronic myeloid leukemia, imatinib resistance, microRNA 451, Polymerase Chain Reaction

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568 Indigenous Adaptation Strategies for Climate Change: Small Farmers’ Options for Sustainable Crop Farming in South-Western Nigeria

Authors: Emmanuel Olasope Bamigboye, Ismail Oladeji Oladosu

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Local people of south-western Nigeria like in other climes, continue to be confronted with the vagaries of changing environments. Through the modification of existing practice and shifting resource base, their strategies for coping with change have enabled them to successfully negotiate the shifts in climate change and the environment. This article analyses indigenous adaptation strategies for climate change with a view to enhancing sustainable crop farming in south –western Nigeria. Multi-stage sampling procedure was used to select 340 respondents from the two major ecological zones (Forest and Derived Savannah) for good geographical spread. The article draws on mixed methods of qualitative research, literature review, field observations, informal interview and multinomial logit regression to capture choice probabilities across the various options of climate change adaptation options among arable crop farmers. The study revealed that most 85.0% of the arable crop farmers were males. It also showed that the use of local climate change adaptation strategies had no relationship with the educational level of the respondents as 77.3% had educational experiences at varying levels. Furthermore, the findings showed that seven local adaptation strategies were commonly utilized by arable crop farmers. Nonetheless, crop diversification, consultation with rainmakers and involvement in non-agricultural ventures were prioritized in the order of 1-3, respectively. Also, multinomial logit analysis result showed that at p ≤ 0.05 level of significance, household size (P<0.08), sex (p<0.06), access to loan(p<0.16), age(p<0.07), educational level (P<0.17) and functional extension contact (P<0.28) were all important in explaining the indigenous climate change adaptation utilized by the arable crops farmers in south-western Nigeria. The study concluded that all the identified local adaptation strategies need to be integrated into the development process for sustainable climate change adaptation.

Keywords: crop diversification, climate change, adaptation option, sustainable, small farmers

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567 Joubert Syndrome in Children as Multicentric Screening in Ten Different Places in World

Authors: Bajraktarevic Adnan, Djukic Branka, Sporisevic Lutvo, Krdzalic Zecevic Belma, Uzicanin Sajra, Hadzimuratovic Admir, Hadzimuratovic Hadzipasic Emina, Abduzaimovic Alisa, Kustric Amer, Suljevic Ismet, Serafi Ismail, Tahmiscija Indira, Khatib Hakam, Semic Jusufagic Aida, Haas Helmut, Vladicic Aleksandra, Aplenc Richard, Kadic Deovic Aida

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Introduction: Joubert syndrome has an autosomal recessive pattern of inheritance. It is referred as the brain malfunctioning and caused due to the underdevelopment of the cerebellar vermis. Associated conditions involving the eye, the kidney, and ocular disease are well described. Aims: Research helps us better understand this diseases, Joubert syndrome and can lead to advances in diagnosis and treatment. Methods: Different several conditions have been described in which the molar tooth sign and characteristics of Joubert syndrome in ten different places in the world. Carrier testing and diagnosis are available if one of these gene mutations has been identified in an affected family member. Results: Authors have described eleven cases during twenty years of Joubert syndrome. It is a clinically and genetically heterogeneous group of disorders characterized by hypoplasia of the cerebellar vermis with the characteristic neuroradiologic molar tooth sign, and accompanying neurologic symptoms, including dysregulation of breathing pattern and developmental delay. We made confirmation of diagnosis in twin sisters with Joubert syndrome with renal anomalies. Ocular symptoms have existed in seven cases (63.64%) from total eleven. Eleven cases were different sex, five boys (45.45%) and six girls (54.44%). Conclusions: Joubert syndrome is inherited as an autosomal recessive genetic disorder with several features of the disease.

Keywords: Joubert syndrome, cerebellooculorenal syndrome, autosomal recessive genetic disorder (ARGD), children

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566 Optimization of SOL-Gel Copper Oxide Layers for Field-Effect Transistors

Authors: Tomas Vincze, Michal Micjan, Milan Pavuk, Martin Weis

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In recent years, alternative materials are gaining attention to replace polycrystalline and amorphous silicon, which are a standard for low requirement devices, where silicon is unnecessarily and high cost. For that reason, metal oxides are envisioned as the new materials for these low-requirement applications such as sensors, solar cells, energy storage devices, or field-effect transistors. Their most common way of layer growth is sputtering; however, this is a high-cost fabrication method, and a more industry-suitable alternative is the sol-gel method. In this group of materials, many oxides exhibit a semiconductor-like behavior with sufficiently high mobility to be applied as transistors. The sol-gel method is a cost-effective deposition technique for semiconductor-based devices. Copper oxides, as p-type semiconductors with free charge mobility up to 1 cm2/Vs., are suitable replacements for poly-Si or a-Si:H devices. However, to reach the potential of silicon devices, a fine-tuning of material properties is needed. Here we focus on the optimization of the electrical parameters of copper oxide-based field-effect transistors by modification of precursor solvent (usually 2-methoxy ethanol). However, to achieve solubility and high-quality films, a better solvent is required. Since almost no solvents have both high dielectric constant and high boiling point, an alternative approach was proposed with blend solvents. By mixing isopropyl alcohol (IPA) and 2-methoxy ethanol (2ME) the precursor reached better solubility. The quality of the layers fabricated using mixed solutions was evaluated in accordance with the surface morphology and electrical properties. The IPA:2ME solution mixture reached optimum results for the weight ratio of 1:3. The cupric oxide layers for optimal mixture had the highest crystallinity and highest effective charge mobility.

Keywords: copper oxide, field-effect transistor, semiconductor, sol-gel method

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565 Rapid Formation of Ortho-Boronoimines and Derivatives for Reversible and Dynamic Bioconjugation Under Physiological Conditions

Authors: Nicholas C. Rose, Christopher D. Spicer

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The regeneration of damaged or diseased tissues would provide an invaluable therapeutic tool in biological research and medicine. Cells must be provided with a number of different biochemical signals in order to form mature tissue through complex signaling networks that are difficult to recreate in synthetic materials. The ability to attach and detach bioactive proteins from material in an iterative and dynamic manner would therefore present a powerful way to mimic natural biochemical signaling cascades for tissue growth. We propose to reversibly attach these bioactive proteins using ortho-boronoimine (oBI) linkages and related derivatives formed by the reaction of an ortho-boronobenzaldehyde with a nucleophilic amine derivative. To enable the use of oBIs for biomaterial modification, we have studied binding and cleavage processes with precise detail in the context of small molecule models. A panel of oBI complexes has been synthesized and screened using a novel Förster resonance energy transfer (FRET) assay, using a cyanine dye FRET pair (Cy3 and Cy5), to identify the most reactive boron-aldehyde/amine nucleophile pairs. Upon conjugation of the dyes, FRET occurs under Cy3 excitation and the resultant ratio of Cy3:Cy5 emission directly correlates to conversion. Reaction kinetics and equilibria can be accurately quantified for reactive pairs, with dissociation constants of oBI derivatives in water (KD) found to span 9-orders of magnitude (10⁻²-10⁻¹¹ M). These studies have provided us with a better understanding of oBI linkages that we hope to exploit to reversibly attach bioconjugates to materials. The long-term aim of the project is to develop a modular biomaterial platform that can be used to help combat chronic diseases such as osteoarthritis, heart disease, and chronic wounds by providing cells with potent biological stimuli for tissue engineering.

Keywords: dynamic, bioconjugation, bornoimine, rapid, physiological

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564 Identification and Quantification of Sesquiterpene Lactones of Sagebrush (Artemisia tridentate) and Its Chemical Modification

Authors: Rosemary Anibogwu, Kavita Sharma, Karl De Jesus

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Sagebrush is an abundant and naturally occurring plant in the Intermountain West region of the United States. The plant contains an array of bioactive compounds such as flavonoids, terpenoids, sterols, and phenolic acids. It is important to identify and characterize these compounds because Native Americans use sagebrush as herbal medicine. These compounds are also utilized for preventing infection in wounds, treating headaches and colds, and possess antitumor properties. This research is an exploratory study on the sesquiterpene present in the leaves of sagebrush. The leaf foliage was extracted with 100 % chloroform and 100 % methanol. The percentage yield for the crude was considerably higher in chloroform. The Thin Layer Chromatography (TLC) analysis of the crude extracted unveiled a brown band at Rf = 0.25 and a dark brown band at Rf = 0.74, along with three unknown faint bands the 254 nm UV lamp. Furthermore, the two distinct brown (Achillin) and dark brown band (Hydroxyachillin) in TLC were further utilized in the isolation of pure compounds with column chromatography. The structures of Achillin and Hydroxyachillin were elucidated based on extensive spectroscopic analysis, including TLC, High-Performance Liquid Chromatography (HPLC), 1D- and 2D-Nuclear Magnetic Resonance (NMR), and Mass Spectroscopy (MS). The antioxidant activities of crude extract and three pure compounds were evaluated in terms of their peroxyl radical scavenging by Ferric Reducing Ability of Plasma (FRAP) and 1,1-Diphenyl-2-picryl-hydrazyl (DPPH) methods. The crude extract showed the antioxidant activity of 18.99 ± 0.51 µmol TEg -1 FW for FRAP and 11.59 ± 0.38 µmol TEg -1 FW for DPPH. The activities of Achillin, Hydroxyachillin, and Quercetagetin trimethyl ether were 13.03, 15.90 and 14.02 µmol TEg -1 FW respectively for the FRAP assay. The three purified compounds have been submitted to the National Cancer Institute 60 cancer cell line for further study.

Keywords: HPLC, nuclear magnetic resonance spectroscopy, sagebrush, sesquiterpene lactones

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