Search results for: gene regulation
2064 Regulating the Emerging Platform Economy in Ethiopia: Issues in the Ride-Hailing Platforms
Authors: Nebiat Lemenih Lenger
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Today, the digital economy is evolving faster than ever in Ethiopia. Platforms that provide a ride-hailing service are growing fast in the country. The market welcomed them as they disrupt it with quality services and lower prices. This revolution is, however, not without challenges. These include cybersecurity breaches, facilitating illegal economic activities, and challenging concepts of privacy. To mitigate the risks and utilize the benefits, appropriate regulation should be introduced in the economy. By identifying legal and institutional gaps in Ethiopia`s digital economy, this research work assists the government`s effort to create a better digital economy. Moreover, this study, being a pioneer study in the area, will be an input for further studies in academia. The research employs a qualitative legal research method and analyzes various legal and policy instruments in Ethiopia in comparison with best international experiences. As this research applies a qualitative research method, a grounded theory method of data analysis is used. The research concluded that Ethiopia is far from designing appropriate legal and regulatory infrastructures. Due to the government monopoly of the sector, there is poor digital infrastructure in the country. The existing labor laws have no specific provisions on the rights and obligations of gig workers.Keywords: Ethiopia, gig economy, digital, ride-hailing, regulation
Procedia PDF Downloads 1002063 Comparison of Rumen Microbial Analysis Pipelines Based on 16s rRNA Gene Sequencing
Authors: Xiaoxing Ye
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To investigate complex rumen microbial communities, 16S ribosomal RNA (rRNA) sequencing is widely used. Here, we evaluated the impact of bioinformatics pipelines on the observation of OTUs and taxonomic classification of 750 cattle rumen microbial samples by comparing three commonly used pipelines (LotuS, UPARSE, and QIIME) with Usearch. In LotuS-based analyses, 189 archaeal and 3894 bacterial OTUs were observed. The observed OTUs for the Usearch analysis were significantly larger than the LotuS results. We discovered 1495 OTUs for archaea and 92665 OTUs for bacteria using Usearch analysis. In addition, taxonomic assignments were made for the rumen microbial samples. All pipelines had consistent taxonomic annotations from the phylum to the genus level. A difference in relative abundance was calculated for all microbial levels, including Bacteroidetes (QIIME: 72.2%, Usearch: 74.09%), Firmicutes (QIIME: 18.3%, Usearch: 20.20%) for the bacterial phylum, Methanobacteriales (QIIME: 64.2%, Usearch: 45.7%) for the archaeal class, Methanobacteriaceae (QIIME: 35%, Usearch: 45.7%) and Methanomassiliicoccaceae (QIIME: 35%, Usearch: 31.13%) for archaeal family. However, the most prevalent archaeal class varied between these two annotation pipelines. The Thermoplasmata was the top class according to the QIIME annotation, whereas Methanobacteria was the top class according to Usearch.Keywords: cattle rumen, rumen microbial, 16S rRNA gene sequencing, bioinformatics pipeline
Procedia PDF Downloads 922062 Regulation of Desaturation of Fatty Acid and Triglyceride Synthesis by Myostatin through Swine-Specific MEF2C/miR222/SCD5 Pathway
Authors: Wei Xiao, Gangzhi Cai, Xingliang Qin, Hongyan Ren, Zaidong Hua, Zhe Zhu, Hongwei Xiao, Ximin Zheng, Jie Yao, Yanzhen Bi
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Myostatin (MSTN) is the master regulator of double muscling phenotype with overgrown muscle and decreased fatness in animals, but its action mode to regulate fat deposition remains to be elucidated. In this study a swin-specific pathway through which MSTN acts to regulate the fat deposition was deciphered. Deep sequenincing of the mRNA and miRNA of fat tissues of MSTN knockout (KO) and wildtype (WT) pigs discovered the positive correlation of myocyte enhancer factor 2C (MEF2C) and fat-inhibiting miR222 expression, and the inverse correlation of miR222 and stearoyl-CoA desaturase 5 (SCD5) expression. SCD5 is rodent-absent and expressed only in pig, sheep and cattle. Fatty acid spectrum of fat tissues revealed a lower percentage of oleoyl-CoA (18:1) and palmitoleyl CoA (16:1) in MSTN KO pigs, which are the catalyzing products of SCD5-mediated desaturation of steroyl CoA (18:0) and palmitoyl CoA (16:0). Blood metrics demonstrated a 45% decline of triglyceride (TG) content in MSTN KO pigs. In light of these observations we hypothesized that MSTN might act through MEF2C/miR222/SCD5 pathway to regulate desaturation of fatty acid as well as triglyceride synthesis in pigs. To this end, real-time PCR and Western blotting were carried out to detect the expression of the three genes stated above. These experiments showed that MEF2C expression was up-regulated by nearly 2-fold, miR222 up-regulated by nearly 3-fold and SCD5 down-regulated by nearly 50% in MSTN KO pigs. These data were consistent with the expression change in deep sequencing analysis. Dual luciferase reporter was then used to confirm the regulation of MEF2C upon the promoter of miR222. Ecotopic expression of MEF2C in preadipocyte cells enhanced miR222 expression by 3.48-fold. CHIP-PCR identified a putative binding site of MEF2C on -2077 to -2066 region of miR222 promoter. Electrophoretic mobility shift assay (EMSA) demonstrated the interaction of MEF2C and miR222 promoter in vitro. These data indicated that MEF2C transcriptionally regulates the expression of miR222. Next, the regulation of miR222 on SCD5 mRNA as well as its physiological consequences were examined. Dual luciferase reporter testing revealed the translational inhibition of miR222 upon the 3´ UTR (untranslated region) of SCD5 in preadipocyte cells. Transfection of miR222 mimics and inhibitors resulted in the down-regulation and up-regulation of SCD5 in preadipocyte cells respectively, consistent with the results from reporter testing. RNA interference of SCD5 in preadipocyte cells caused 26.2% reduction of TG, in agreement with the results of TG content in MSTN KO pigs. In summary, the results above supported the existence of a molecular pathway that MSTN signals through MEF2C/miR222/SCD5 to regulate the fat deposition in pigs. This swine-specific pathway offers potential molecular markers for the development and breeding of a new pig line with optimised fatty acid composition. This would benefit human health by decreasing the takeup of saturated fatty acid.Keywords: fat deposition, MEF2C, miR222, myostatin, SCD5, pig
Procedia PDF Downloads 1332061 Variable Shunt Reactors for Reactive Power Compensation of HV Subsea Cables
Authors: Saeed A. AlGhamdi, Nabil Habli, Vinoj Somasanran
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This paper presents an application of 230 kV Variable Shunt Reactors (VSR) used to compensate reactive power of dual 90 KM subsea cables. VSR integrates an on-load tap changer (OLTC) that adjusts reactive power compensation to maintain acceptable bus voltages under variable load profile and network configuration. An automatic voltage regulator (AVR) or a power management system (PMS) that allows VSR rating to be changed in discrete steps typically controls the OLTC. Typical regulation range start as minimum as 20% up to 100% and are available for systems up to 550kV. The regulation speed is normally in the order of seconds per step and approximately a minute from maximum to minimum rating. VSR can be bus or line connected depending on line/cable length and compensation requirements. The flexible reactive compensation ranges achieved by recent VSR technologies have enabled newer facilities design to deploy line connected VSR through either disconnect switches, which saves space and cost, or through circuit breakers. Lines with VSR are typically energized with lower taps (reduced reactive compensation) to minimize or remove the presence of delayed zero crossing.Keywords: power management, reactive power, subsea cables, variable shunt reactors
Procedia PDF Downloads 2572060 Precise Identification of Clustered Regularly Interspaced Short Palindromic Repeats-Induced Mutations via Hidden Markov Model-Based Sequence Alignment
Authors: Jingyuan Hu, Zhandong Liu
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CRISPR genome editing technology has transformed molecular biology by accurately targeting and altering an organism’s DNA. Despite the state-of-art precision of CRISPR genome editing, the imprecise mutation outcome and off-target effects present considerable risk, potentially leading to unintended genetic changes. Targeted deep sequencing, combined with bioinformatics sequence alignment, can detect such unwanted mutations. Nevertheless, the classical method, Needleman-Wunsch (NW) algorithm may produce false alignment outcomes, resulting in inaccurate mutation identification. The key to precisely identifying CRISPR-induced mutations lies in determining optimal parameters for the sequence alignment algorithm. Hidden Markov models (HMM) are ideally suited for this task, offering flexibility across CRISPR systems by leveraging forward-backward algorithms for parameter estimation. In this study, we introduce CRISPR-HMM, a statistical software to precisely call CRISPR-induced mutations. We demonstrate that the software significantly improves precision in identifying CRISPR-induced mutations compared to NW-based alignment, thereby enhancing the overall understanding of the CRISPR gene-editing process.Keywords: CRISPR, HMM, sequence alignment, gene editing
Procedia PDF Downloads 592059 The Political and Academic Consideration of Unregulated Concept of Rome Statute in Law No. 26 Year 2000 about Indonesia’s Human Right Court
Authors: Muhammad Iqbal Rachman, Mohammad Faisol Soleh
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The Law No. 26 Year 2000 about Indonesia’s Human Right Court became a new legal enforcement frame of human right law in Indonesia. The new spirit based on some international propulsion in order to enforce human right which basic right of everyone that appearance since in fetus. This matters indicated how crucial the arrangement of human right law, considering the role of state on human right enforcement in this context which became main pillar or instrument to accommodate citizen interest. Basically, the adopting of Law No. 26 Year 2000 came from the womb of concept international crimes regulation based on Rome Statute which became the international law instrument in order to legal enforce of international crimes. But in the other side, the enactment Rome Statute concept in Indonesia has facing with political and academics interest which resulted unaccommodating every type of international crimes in Law No. 26 Year 2000. The analyzing of political and academics background became the fundamental point to find out the solutions based on the regulation of Rome Statute concept matters in Indonesia.Keywords: academic consideration, human right, political consideration, rome statute, unregulated concept
Procedia PDF Downloads 2902058 Proinflammatory Response of Agglomerated TiO2 Nanoparticles in Human-Immune Cells
Authors: Vaiyapuri Subbarayn Periasamy, Jegan Athinarayanan, Ali A. Alshatwi
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The widespread use of Titanium oxide nanoparticles (TiO2-NPs), now are found with different physic-chemical properties (size, shape, chemical properties, agglomeration, etc.) in many processed foods, agricultural chemicals, biomedical products, food packaging and food contact materials, personal care products, and other consumer products used in daily life. Growing evidences have been highlighted that there are risks of physico-chemical properties dependent toxicity with special attention to “TiO2-NPs and human immune system”. Unfortunately, agglomeration and aggregation have frequently been ignored in immuno-toxicological studies, even though agglomeration and aggregation would be expected to affect nanotoxicity since it changes the size, shape, surface area, and other properties of the TiO2-NPs. In this present investigation, we assessed the immune toxic effect of TiO2-NPs on human immune cells Total WBC including Lymphocytes (T cells (CD3+), T helper cells (CD3+, CD4+), Suppressor/cytotoxic T cells (CD3+/CD8+) and NK cells (CD3-/CD16+ and CD56+), Monocytes (CD14+, CD3-) and B lymphocytes (CD19+, CD3-) in order to find the immunological response (IL1A, IL1B, IL2 IL-4, IL5 IL-6, IL-10, IL-12, IL-13, IFN-γ, TGF-β, and TNF-a) and redox gene regulation (TNF, p53, BCl-2, CAT, GSTA4, TNF, CYP1A, POR, SOD1, GSTM3, GPX1, and GSR1)-linking physicochemical properties with special reference to agglomeration of TiO2-NPs. Our findings suggest that TiO2-NPs altered cytokine production, enhanced phagocytic indexing, metabolic stress through specific immune regulatory- genes expression in different WBC subsets and may contribute to pro-inflammatory response. Although TiO2-NPs have great advantages in the personal care products, biomedical, food and agricultural products, its chronic and acute immune-toxicity still need to be assessed carefully with special reference to food and environmental safety.Keywords: TiO2 nanoparticles, oxidative stress, cytokine, human immune cells
Procedia PDF Downloads 3992057 Thermal Regulation of Channel Flows Using Phase Change Material
Authors: Kira Toxopeus, Kamran Siddiqui
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Channel flows are common in a wide range of engineering applications. In some types of channel flows, particularly the ones involving chemical or biological processes, the control of the flow temperature is crucial to maintain the optimal conditions for the chemical reaction or to control the growth of biological species. This often becomes an issue when the flow experiences temperature fluctuations due to external conditions. While active heating and cooling could regulate the channel temperature, it may not be feasible logistically or economically and is also regarded as a non-sustainable option. Thermal energy storage utilizing phase change material (PCM) could provide the required thermal regulation sustainably by storing the excess heat from the channel and releasing it back as required, thus regulating the channel temperature within a range in the proximity of the PCM melting temperature. However, in designing such systems, the configuration of the PCM storage within the channel is critical as it could influence the channel flow dynamics, which would, in turn, affect the heat exchange between the channel fluid and the PCM. The present research is focused on the investigation of the flow dynamical behavior in the channel during heat transfer from the channel flow to the PCM thermal energy storage. Offset vertical columns in a narrow channel were used that contained the PCM. Two different column shapes, square and circular, were considered. Water was used as the channel fluid that entered the channel at a temperature higher than that of the PCM melting temperature. Hence, as the water was passing through the channel, the heat was being transferred from the water to the PCM, causing the PCM to store the heat through a phase transition from solid to liquid. Particle image velocimetry (PIV) was used to measure the two-dimensional velocity field of the channel flow as it flows between the PCM columns. Thermocouples were also attached to the PCM columns to measure the PCM temperature at three different heights. Three different water flow rates (0.5, 0.75 and 1.2 liters/min) were considered. At each flow rate, experiments were conducted at three different inlet water temperatures (28ᵒC, 33ᵒC and 38ᵒC). The results show that the flow rate and the inlet temperature influenced the flow behavior inside the channel.Keywords: channel flow, phase change material, thermal energy storage, thermal regulation
Procedia PDF Downloads 1452056 Molecular Detection and Characterization of Infectious Bronchitis Virus from Libya
Authors: Abdulwahab Kammon, Tan Sheau Wei, Abdul Rahman Omar, Abdunaser Dayhum, Ibrahim Eldghayes, Monier Sharif
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Infectious bronchitis virus (IBV) is a very dynamic and evolving virus which causing major economic losses to the global poultry industry. Recently, the Libyan poultry industry faced severe outbreak of respiratory distress associated with high mortality and dramatic drop in egg production. Tracheal and cloacal swabs were analyzed for several poultry viruses. IBV was detected using SYBR Green I real-time PCR detection based on the nucleocapsid (N) gene. Sequence analysis of the partial N gene indicated high similarity (~ 94%) to IBV strain 3382/06 that was isolated from Taiwan. Even though the IBV strain 3382/06 is more similar to that of the Mass type H120, the isolate has been implicated associated with intertypic recombinant of 3 putative parental IBV strains namely H120, Taiwan strain 1171/92 and China strain CK/CH/LDL/97I. Complete sequencing and antigenicity studies of the Libya IBV strains are currently underway to determine the evolution of the virus and its importance in vaccine induced immunity. In this paper, we documented for the first time the presence of possibly variant IBV strain from Libya which required a dramatic change in the vaccination program.Keywords: Libya, infectious bronchitis, molecular characterization, viruses, vaccine
Procedia PDF Downloads 4752055 Aerobic Biodegradation of a Chlorinated Hydrocarbon by Bacillus Cereus 2479
Authors: Srijata Mitra, Mobina Parveen, Pranab Roy, Narayan Chandra Chattopadhyay
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Chlorinated hydrocarbon can be a major pollution problem in groundwater as well as soil. Many people interact with these chemicals on daily accidentally or by professionally in the laboratory. One of the most common sources for Chlorinated hydrocarbon contamination of soil and groundwater are industrial effluents. The wide use and discharge of Trichloroethylene (TCE), a volatile chlorohydrocarbon from chemical industry, led to major water pollution in rural areas. TCE is an mainly used as an industrial metal degreaser in industries. Biotransformation of TCE to the potent carcinogen vinyl chloride (VC) by consortia of anaerobic bacteria might have role for the above purpose. For these reasons, the aim of current study was to isolate and characterized the genes involved in TCE metabolism and also to investigate the in silico study of those genes. To our knowledge, only one aromatic dioxygenase system, the toluene dioxygenase in Pseudomonas putida F1 has been shown to be involved in TCE degradation. This is first instance where Bacillus cereus group being used in biodegradation of trichloroethylene. A novel bacterial strain 2479 was isolated from oil depot site at Rajbandh, Durgapur (West Bengal, India) by enrichment culture technique. It was identified based on polyphasic approach and ribotyping. The bacterium was gram positive, rod shaped, endospore forming and capable of degrading trichloroethylene as the sole carbon source. On the basis of phylogenetic data and Fatty Acid Methyl Ester Analysis, strain 2479 should be placed within the genus Bacillus and species cereus. However, the present isolate (strain 2479) is unique and sharply different from the usual Bacillus strains in its biodegrading nature. Fujiwara test was done to estimate that the strain 2479 could degrade TCE efficiently. The gene for TCE biodegradation was PCR amplified from genomic DNA of Bacillus cereus 2479 by using todC1 gene specific primers. The 600bp amplicon was cloned into expression vector pUC I8 in the E. coli host XL1-Blue and expressed under the control of lac promoter and nucleotide sequence was determined. The gene sequence was deposited at NCBI under the Accession no. GU183105. In Silico approach involved predicting the physico-chemical properties of deduced Tce1 protein by using ProtParam tool. The tce1 gene contained 342 bp long ORF encoding 114 amino acids with a predicted molecular weight 12.6 kDa and the theoretical pI value of the polypeptide was 5.17, molecular formula: C559H886N152O165S8, total number of atoms: 1770, aliphatic index: 101.93, instability index: 28.60, Grand Average of Hydropathicity (GRAVY): 0.152. Three differentially expressed proteins (97.1, 40 and 30 kDa) were directly involved in TCE biodegradation, found to react immunologically to the antibodies raised against TCE inducible proteins in Western blot analysis. The present study suggested that cloned gene product (TCE1) was capable of degrading TCE as verified chemically.Keywords: cloning, Bacillus cereus, in silico analysis, TCE
Procedia PDF Downloads 4032054 Board Regulation and Its Impact on Composition and Effects: Evidence from German Cooperative Banks
Authors: Markus Stralla
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This study employs a GMM framework to examine the impact of potential regulatory intervention regarding the occupations of supervisory board members in cooperative banking. To achieve insights, the study proceeds in two different ways. First, it investigates the changes in board structure prior and following to the German Act to Strengthen Financial Market and Insurance Supervision (FinVAG). Second, the study estimates the influence of Ph.D.Share, professional concentration and supervisory power on bank-risk changes in consideration of the implementation of FinVAG. Therefore, the study is based on a sample of 246 German cooperative banks from 2006-2011 while applying four different measures of bank risk, namely credit-, equity-, liquidity-risk, and Z-Score, with the former three also being addressed in FinVAG. Results indicate that the implementation of FinVAG results in (most likely unintentional) structural changes, especially at the expense of farmers, and affects all risk measures and relations between risk measures and supervisory board characteristics in a risk-reducing and therefore intended way. To disentangle the complex relationship between board characteristics and risk measures, the study utilizes two-step system GMM estimator to account for unobserved heterogeneity and simultaneity in order to reduce endogeneity problems. The findings may be especially relevant for stakeholders, regulators, supervisors and managers.Keywords: bank governance, bank risk-taking, board of directors, regulation
Procedia PDF Downloads 4312053 Influence of the Quality of the Recycled Aggregates in Concrete Pavement
Authors: Viviana Letelier, Ester Tarela, Bianca Lopez, Pedro Muñoz, Giacomo Moriconi
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The environmental impact has become a global concern during the last decades. Several alternatives have been proposed and studied to minimize this impact in different areas. The reuse of aggregates from old concretes to manufacture new ones not only can reduce this impact but is also a way to optimize the resource management. The effect of the origin of the reused aggregates from two different origin materials in recycled concrete pavement is studied here. Using the dosing applied by a pavement company, coarse aggregates in the 6.3-25 mm fraction are replaced by recycled aggregates with two different origins: old concrete pavements with similar origin strength to the one of the control concrete, and precast concrete pipes with smaller strengths than the one of the control concrete. The replacement percentages tested are 30%, 40% and 50% in both cases. The compressive strength tests are performed after 7, 14, 28 and 90 curing days, the flexural strength tests and the elasticity modulus tests after 28 and 90 curing days. Results show that the influence of the quality of the origin concrete in the mechanical properties of recycled concretes is not despicable. Concretes with up to a 50% of recycled aggregates from the concrete pavement have similar compressive strengths to the ones of the control concrete and slightly smaller flexural strengths that, however, in all cases exceed the minimum of 5MPa after 28 curing days stablished by the Chilean regulation for pavement concretes. On the other hand, concretes with recycled aggregates from precast concrete pipes show significantly lower compressive strengths after 28 curing days. The differences with the compressive strength of the control concrete increase with the percentage of replacement, reaching a 13% reduction when 50% of the aggregates are replaced. The flexural strength also suffers significant reductions that increase with the percentage of replacement, only obeying the Chilean regulation when 30% of the aggregates are recycled after 28 curing days. Nevertheless, after 90 curing days, all series obey the regulation requirements. Results show, not only the importance of the quality of the origin concrete, but also the significance of the curing days, that may allow the use of less quality recycled material without important strength losses.Keywords: flexural strength of recycled concrete., mechanical properties of recycled concrete, recycled aggregates, recycled concrete pavements
Procedia PDF Downloads 2542052 The First Complete Mitochondrial Genome of Melon Thrips, Thrips palmi (Thripinae: Thysanoptera): Vector for Tospoviruses
Authors: Kaomud Tyagi, Rajasree Chakraborty, Shantanu Kundu, Devkant Singha, Kailash Chandra, Vikas Kumar
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The melon thrips, Thrips palmi is a serious pest of a wide range of agriculture crops and also act as vectors for plant viruses (genus Tospovirus, family Bunyaviridae). More molecular data on this species is required to understand the cryptic speciation and evolutionary affiliations. Mitochondrial genomes have been widely used in phylogenetic and evolutionary studies in insect. So far, mitogenomes of five thrips species (Anaphothrips obscurus, Frankliniella intonsa, Frankliniella occidentalis, Scirtothrips dorsalis and Thrips imaginis) is available in the GenBank database. In this study, we sequenced the first complete mitogenome T. palmi and compared it with available thrips mitogenomes. We assembled the mitogenome from the whole genome sequencing data generated using Illumina Hiseq2500. Annotation was performed using MITOS web-server to estimate the location of protein coding genes (PCGs), transfer RNA (tRNAs), ribosomal RNAs (rRNAs) and their secondary structures. The boundaries of PCGs and rRNAs was confirmed manually in NCBI. Phylogenetic analyses were performed using the 13 PCGs data using maximum likelihood (ML) in PAUP, and Bayesian inference (BI) in MrBayes 3.2. The complete mitogenome of T. palmi was 15,333 base pairs (bp), which was greater than the genomes of A. obscurus (14,890bp), F. intonsa (15,215 bp), F. occidentalis (14,889 bp) and S. dorsalis South Asia strain (SA1) (14,283 bp), but smaller than the genomes of T. imaginis (15,407 bp) and S. dorsalis East Asia strain (EA1) (15,343bp). Like in other thrips species, the mitochondrial genome of T. palmi was represented by 37 genes, including 13 PCGs, large and small ribosomal RNA (rrnL and rrnS) genes, 22 transfer RNA (tRNAs) genes (with one extra gene for trn-Serine) and two A+T-rich control regions (CR1 and CR2). Thirty one genes were observed on heavy (H) strand and six genes on the light (L) strand. The six tRNA genes (trnG,trnK, trnY, trnW, trnF, and trnH) were found to be conserved in all thrips species mitogenomes in their locations relative to a protein-coding or rRNA gene upstream or downstream. The gene arrangements of T. palmi is very close to T. imaginis except the rearrangements in tRNAs genes: trnR (arginine), and trnE (glutamic acid) were found to be located between cox3 and CR2 in T. imaginis which were translocated between atp6 and CR1 in T. palmi; trnL1 (Leucine) and trnS1(Serine) were located between atp6 and CR1 in T. imaginis which were translocated between cox3 and CR2 in T. palmi. The location of CR1 upstream of nad5 gene was suggested to be ancestral condition of the thrips species in subfamily Thripinae, was also observed in T. palmi. Both the Maximum likelihood (ML) and Bayesian Inference (BI) phylogenetic trees generated resulted in similar topologies. The T. palmi was clustered with T. imaginis. We concluded that more molecular data on the diverse thrips species from different hierarchical level is needed, to understand the phylogenetic and evolutionary relationships among them.Keywords: thrips, comparative mitogenomics, gene rearrangements, phylogenetic analysis
Procedia PDF Downloads 1722051 Boosting Economic Value in Ghana’s Film Industry: Rethinking Media Policy, Regulation and Copyright Law
Authors: Sela Adjei
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This paper aims to rationalize the need for media policy implementation and copyright enforcement to address various challenges faced within Ghana’s film industry. After Ghana transitioned to democratic rule in 1992, critics and media professionals advocated a national media policy. This advocacy subsequently resulted in agitation for media deregulation and loosening of grip on state-owned media organizations. The reinstatement of constitutional rule in 1992 paved the way for the state to lax its monopoly of the media within the democratic context of a free market economy. The National Media Commission proposed a media policy and broadcast bill which was presented to parliament but has still not been passed into law. This legislative lapse partly contributed to the influx of unregulated foreign content. Accessible foreign media content subsequently promoted a system of unfair competition that radically undermined locally produced content, putting a generation of thriving film producers out of work. Drawing on reflections from a series of structured interviews, focus group discussions and creative workshops, the findings of this study maintain that the various challenges confronting Ghanaian filmmakers is centred around inadequate funding opportunities, copyright violation and policy implementation issues. Using the film industry structure and value chain analysis, the various challenges faced by the selected film producers were discussed and critically analyzed. A significant aspect of this study is the solution-driven approach adopted in outlining the practical recommendations that will boost the aesthetic, cultural and economic value of Ghanaian film productions. Based on the discussions and conclusions drawn with the various stakeholders within Ghana’s creative industries, the paper makes a strong case for firm state regulation, copyright enforcement and policy implementation to grow Ghana’s film industry.Keywords: film, value, copyright, media, policy, culture, regulation, economy
Procedia PDF Downloads 732050 Anti-Obesity Effects of Pteryxin in Peucedanum japonicum Thunb Leaves through Different Pathways of Adipogenesis In-Vitro
Authors: Ruwani N. Nugara, Masashi Inafuku, Kensaku Takara, Hironori Iwasaki, Hirosuke Oku
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Pteryxin from the partially purified hexane phase (HP) of Peucedanum japonicum Thunb (PJT) was identified as the active compound related to anti-obesity. Thus, in this study we investigated the mechanisms related to anti-obesity activity in-vitro. The HP was fractionated, and effect on the triglyceride (TG) content was evaluated in 3T3-L1 and HepG2 cells. Comprehensive spectroscopic analyses were used to identify the structure of the active compound. The dose dependent effect of active constituent on the TG content, and the gene expressions related to adipogenesis, fatty acid catabolism, energy expenditure, lipolysis and lipogenesis (20 μg/mL) were examined in-vitro. Furthermore, higher dosage of pteryxin (50μg/mL) was tested against 20μg/mL in 3T3-L1 adipocytes. The mRNA were subjected to SOLiD next generation sequencer and the obtained data were analyzed by Ingenuity Pathway Analysis (IPA). The active constituent was identified as pteryxin, a known compound in PJT. However, its biological activities against obesity have not been reported previously. Pteryxin dose dependently suppressed TG content in both 3T3-L1 adipocytes and HepG2 hepatocytes (P < 0.05). Sterol regulatory element-binding protein-1 (SREBP1 c), Fatty acid synthase (FASN), and acetyl-CoA carboxylase-1 (ACC1) were downregulated in pteryxin-treated adipocytes (by 18.0, 36.1 and 38.2%; P < 0.05, respectively) and hepatocytes (by 72.3, 62.9 and 38.8%, respectively; P < 0.05) indicating its suppressive effects on fatty acid synthesis. The hormone-sensitive lipase (HSL), a lipid catabolising gene was upregulated (by 15.1%; P < 0.05) in pteryxin-treated adipocytes suggesting improved lipolysis. Concordantly, the adipocyte size marker gene, paternally expressed gene1/mesoderm specific transcript (MEST) was downregulated (by 42.8%; P < 0.05), further accelerating the lipolytic activity. The upregulated trend of uncoupling protein 2 (UCP2; by 77.5%; P < 0.05) reflected the improved energy expenditure due to pteryxin. The 50μg/mL dosage of pteryxin completely suppressed PPARγ, MEST, SREBP 1C, HSL, Adiponectin, Fatty Acid Binding Protein (FABP) 4, and UCP’s in 3T3-L1 adipocytes. The IPA suggested that pteryxin at 20μg/mL and 50μg/mL suppress obesity in two different pathways, whereas the WNT signaling pathway play a key role in the higher dose of pteryxin in preadipocyte stage. Pteryxin in PJT play the key role in regulating lipid metabolism related gene network and improving energy production in vitro. Thus, the results suggests pteryxin as a new natural compound to be used as an anti-obesity drug in pharmaceutical industry.Keywords: obesity, peucedanum japonicum thunb, pteryxin, food science
Procedia PDF Downloads 4562049 In vitro Modeling of Aniridia-Related Keratopathy by the Use of Crispr/Cas9 on Limbal Epithelial Cells and Rescue
Authors: Daniel Aberdam
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Haploinsufficiency of PAX6 in humans is the main cause of congenital aniridia, a rare eye disease characterized by reduced visual acuity. Patients have also progressive disorders including cataract, glaucoma and corneal abnormalities making their condition very challenging to manage. Aniridia-related keratopathy (ARK), caused by a combination of factors including limbal stem-cell deficiency, impaired healing response, abnormal differentiation, and infiltration of conjunctival cells onto the corneal surface, affects up to 95% of patients. It usually begins in the first decade of life resulting in recurrent corneal erosions, sub-epithelial fibrosis with corneal decompensation and opacification. Unfortunately, current treatment options for aniridia patients are currently limited. Although animal models partially recapitulate this disease, there is no in vitro cellular model of AKT needed for drug/therapeutic tools screening and validation. We used genome editing (CRISPR/Cas9 technology) to introduce a nonsense mutation found in patients into one allele of the PAX6 gene into limbal stem cells. Resulting mutated clones, expressing half of the amount of PAX6 protein and thus representative of haploinsufficiency were further characterized. Sequencing analysis showed that no off-target mutations were induced. The mutated cells displayed reduced cell proliferation and cell migration but enhanced cell adhesion. Known PAX6 targets expression was also reduced. Remarkably, addition of soluble recombinant PAX6 protein into the culture medium was sufficient to activate endogenous PAX6 gene and, as a consequence, rescue the phenotype. It strongly suggests that our in vitro model recapitulates well the epithelial defect and becomes a powerful tool to identify drugs that could rescue the corneal defect in patients. Furthermore, we demonstrate that the homeotic transcription factor Pax6 is able to be uptake naturally by recipient cells to function into the nucleus.Keywords: Pax6, crispr/cas9, limbal stem cells, aniridia, gene therapy
Procedia PDF Downloads 2102048 Fundamental Theory of the Evolution Force: Gene Engineering utilizing Synthetic Evolution Artificial Intelligence
Authors: L. K. Davis
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The effects of the evolution force are observable in nature at all structural levels ranging from small molecular systems to conversely enormous biospheric systems. However, the evolution force and work associated with formation of biological structures has yet to be described mathematically or theoretically. In addressing the conundrum, we consider evolution from a unique perspective and in doing so we introduce the “Fundamental Theory of the Evolution Force: FTEF”. We utilized synthetic evolution artificial intelligence (SYN-AI) to identify genomic building blocks and to engineer 14-3-3 ζ docking proteins by transforming gene sequences into time-based DNA codes derived from protein hierarchical structural levels. The aforementioned served as templates for random DNA hybridizations and genetic assembly. The application of hierarchical DNA codes allowed us to fast forward evolution, while dampening the effect of point mutations. Natural selection was performed at each hierarchical structural level and mutations screened using Blosum 80 mutation frequency-based algorithms. Notably, SYN-AI engineered a set of three architecturally conserved docking proteins that retained motion and vibrational dynamics of native Bos taurus 14-3-3 ζ.Keywords: 14-3-3 docking genes, synthetic protein design, time-based DNA codes, writing DNA code from scratch
Procedia PDF Downloads 1182047 Evaluation of Immune Responses of Gamma-Irradiated, Electron Beam Irradiated FMD Virus Type O/IRN/2007 Vaccines and DNA Vaccine- Based on the VP1 Gene by a Prime-Boost Strategy in a Mouse Model
Authors: Farahnaz Motamedi Sedeh, Homayoon Mahravani, Parvin Shawrang, Mehdi Behgar
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Most countries use inactivated binary ethylenimine (BEI) vaccines to control and prevent Foot-and-Mouth Disease (FMD). However, this vaccine induces a short-term humoral immune response in animals. This study investigated the cellular and humoral immune responses in homologous and prime-boost (PB) groups in the BALB/c mouse model. FMDV strain O/IRN/1/2007 was propagated in the BHK-21 cell line and inactivated by three methods, including a chemical with BEI to produce a conventional vaccine (CV), a gamma irradiation vaccine (GIV), and an electron irradiated vaccine (EIV). Three vaccines were formulated with the adjuvant aluminum hydroxide gel. In addition, a DNA vaccine was prepared by amplifying the virus VP1 gene pcDNA3.1 plasmid. In addition, the plasmid encoding the granulocyte-macrophage colony-stimulating factor gene (GM-CSF) was used as a molecular adjuvant. Eleven groups of five mice each were selected, and the vaccines were administered as homologous and heterologous strategy prime-boost (PB) in three doses two weeks apart. After the evaluation of neutralizing antibodies, interleukin (IL)-2, IL-4, IL-10, interferon-gamma (INF-γ), and MTT assays were compared in the different groups. The pcDNA3.1+VP1 cassette was prepared and confirmed as a DNA vaccine. The virus was inactivated by gamma rays and electron beams at 50 and 55 kGy as GIV and EIV, respectively. Splenic lymphocyte proliferation in the inactivated vaccinated homologous groups was significantly lower (P≤0.05) compared with the heterologous prime-boosts (PB1, PB2, PB3) and DNA + GM-CSF groups (P≤0.05). The highest SNT titer was observed in the inactivated vaccine groups. IFN-γ and IL-2 were higher in the vaccinated groups. It was found that although there was a protective humoral immune response in the groups with inactivated vaccine, there was adequate cellular immunity in the group with the DNA vaccine. However, the strongest cellular and humoral immunity was observed in the PB groups. The primary injection was accompanied by DNA vaccine + GM-CSF and boosted injection with GIV or CV.Keywords: foot and mouth disease, irradiated vaccine, immune responses, DNA vaccine, prime boost strategy
Procedia PDF Downloads 222046 BRG1 and Ep300 as a Transcriptional Regulators of Breast Cancer Growth
Authors: Maciej Sobczak, Julita Pietrzak, Tomasz Płoszaj, Agnieszka Robaszkiewicz
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Brg1, a member of SWI/SNF complex, plays a role in chromatin remodeling, therefore, regulates expression of many genes. Brg1 is an ATPase of SWI/SNF complex, thus its activity requires ATP. Through its bromodomain recognizes acetylated histone residues and evicts them, thus promoting transcriptionally active state of chromatin. One of the enzymes that is responsible for acetylation of histone residues is Ep300. It was previously shown in the literature that cooperation of Brg1 and Ep300 occurs at the promoter regions that have binding sites for E2F-family transcription factors as well as CpG islands. According to literature, approximately 20% of human cancer possess mutation in Brg1 or any other crucial SWI/SNF subunit. That phenomenon makes Brg1-Ep300 a very promising target for anti-cancer therapy. Therefore in our study, we investigated if physical interaction between Brg1 and Ep300 exists and what impact those two proteins have on key for breast cancer cells processes such as DNA damage repair and cell proliferation. Bioinformatical analysis pointed out, that genes involved in cell proliferation and DNA damage repair are overexpressed in MCF7 and MDA-MB-231 cells. Moreover, promoter regions of these genes are highly acetylated, which suggests high transcriptional activity of those sites. Notably, many of those gene possess within their promoters an E2F, Brg1 motives, as well as CpG islands and acetylated histones. Our data show that Brg1 physically interacts with Ep300, and together they regulate expression of genes involved in DNA damage repair and cell proliferation. Upon inhibiting Brg1 or Ep300, expression of vital for cancer cell survival genes such as CDK2/4, BRCA1/2, PCNA, and XRCC1 is decreased in MDA-MB-231 and MCF7 cells. Moreover, inhibition or silencing of either Brg1 or Ep300 leads to cell cycle arrest in G1. After inhibition of BRG1 or Ep300 on tested gene promoters, the repressor complex including Rb, HDAC1, and EZH2 is formed, which inhibits gene expression. These results highlight potentially significant target for targeted anticancer therapy to be introduced as a supportive therapy.Keywords: brg1, ep300, breast cancer, epigenetics
Procedia PDF Downloads 1882045 Identification of New Familial Breast Cancer Susceptibility Genes: Are We There Yet?
Authors: Ian Campbell, Gillian Mitchell, Paul James, Na Li, Ella Thompson
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The genetic cause of the majority of multiple-case breast cancer families remains unresolved. Next generation sequencing has emerged as an efficient strategy for identifying predisposing mutations in individuals with inherited cancer. We are conducting whole exome sequence analysis of germ line DNA from multiple affected relatives from breast cancer families, with the aim of identifying rare protein truncating and non-synonymous variants that are likely to include novel cancer predisposing mutations. Data from more than 200 exomes show that on average each individual carries 30-50 protein truncating mutations and 300-400 rare non-synonymous variants. Heterogeneity among our exome data strongly suggest that numerous moderate penetrance genes remain to be discovered, with each gene individually accounting for only a small fraction of families (~0.5%). This scenario marks validation of candidate breast cancer predisposing genes in large case-control studies as the rate-limiting step in resolving the missing heritability of breast cancer. The aim of this study is to screen genes that are recurrently mutated among our exome data in a larger cohort of cases and controls to assess the prevalence of inactivating mutations that may be associated with breast cancer risk. We are using the Agilent HaloPlex Target Enrichment System to screen the coding regions of 168 genes in 1,000 BRCA1/2 mutation-negative familial breast cancer cases and 1,000 cancer-naive controls. To date, our interim analysis has identified 21 genes which carry an excess of truncating mutations in multiple breast cancer families versus controls. Established breast cancer susceptibility gene PALB2 is the most frequently mutated gene (13/998 cases versus 0/1009 controls), but other interesting candidates include NPSR1, GSN, POLD2, and TOX3. These and other genes are being validated in a second cohort of 1,000 cases and controls. Our experience demonstrates that beyond PALB2, the prevalence of mutations in the remaining breast cancer predisposition genes is likely to be very low making definitive validation exceptionally challenging.Keywords: predisposition, familial, exome sequencing, breast cancer
Procedia PDF Downloads 4972044 Artificial Intelligent Tax Simulator to Minimize Tax Liability for Multinational Corporations
Authors: Sean Goltz, Michael Mayo
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The purpose of this research is to use Global-Regulation.com database of the world laws, focusing on tax treaties between countries, in order to create an AI-driven tax simulator that will run an AI agent through potential tax scenarios across countries. The AI agent goal is to identify the scenario that will result in minimum tax liability based on tax treaties between countries. The results will be visualized by a three dimensional matrix. This will be an online web application. Multinational corporations are running their business through multiple countries. These countries, in turn, have a tax treaty with many other countries to regulate the payment of taxes on income that is transferred between these countries. As a result, planning the best tax scenario across multiple countries and numerous tax treaties is almost impossible. This research propose to use Global-Regulation.com database of word laws in English (machine translated by Google and Microsoft API’s) in order to create a simulator that will include the information in the tax treaties. Once ready, an AI agent will be sent through the simulator to identify the scenario that will result in minimum tax liability. Identifying the best tax scenario across countries may save multinational corporations, like Google, billions of dollars annually. Given the nature of the raw data and the domain of taxes (i.e., numbers), this is a promising ground to employ artificial intelligence towards a practical and beneficial purpose.Keywords: taxation, law, multinational, corporation
Procedia PDF Downloads 2022043 Challenge of Baseline Hydrology Estimation at Large-Scale Watersheds
Authors: Can Liu, Graham Markowitz, John Balay, Ben Pratt
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Baseline or natural hydrology is commonly employed for hydrologic modeling and quantification of hydrologic alteration due to manmade activities. It can inform planning and policy related efforts for various state and federal water resource agencies to restore natural streamflow flow regimes. A common challenge faced by hydrologists is how to replicate unaltered streamflow conditions, particularly in large watershed settings prone to development and regulation. Three different methods were employed to estimate baseline streamflow conditions for 6 major subbasins the Susquehanna River Basin; those being: 1) incorporation of consumptive water use and reservoir operations back into regulated gaged records; 2) using a map correlation method and flow duration (exceedance probability) regression equations; 3) extending the pre-regulation streamflow records based on the relationship between concurrent streamflows at unregulated and regulated gage locations. Parallel analyses were perform among the three methods and limitations associated with each are presented. Results from these analyses indicate that generating baseline streamflow records at large-scale watersheds remain challenging, even with long-term continuous stream gage records available.Keywords: baseline hydrology, streamflow gage, subbasin, regression
Procedia PDF Downloads 3272042 Regulation of PKA-Dependent Calcineurin as a Switch in Cell Secretion
Authors: Hani M. M. Alothaid, Louise Robson, Richmond Muimo
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This study will investigate cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA) dependent calcineurin (Cn), known as protein phosphatase 2 B (PP2B) as well, regulation of chloride ion (Cl⁻) secretion and the release of pro-inflammatory molecules in immune cells such as cytokines. THP-1-derived monocytes, primary human monocytes and the bronchial epithelial cell line (16HBE14o-) were used in this study. The 16HBE14o- cells were chosen as positive control. Hence, to further confirm the expression of cystic fibrosis transmembrane conductance regulator (CFTR), calcium binding protein (S100A10), annexin A2 (AnxA2) and calcineurin A subunit (CnA) in all three cell types, cell lysate was probed against corresponding primary antibodies by immunoblotting. Western blot analyses show the expression of CFTR, AnxA2, CnA and S100A10 in THP-1-derived monocytes and primary human monocytes. In conclusion, CFTR, S100A10, CnA and AnxA2 are expressed in THP-1-derived monocytes and primary human monocytes and regulate Cl⁻ secretion. Also, they may play a role in the pro-inflammatory molecules release. The ongoing work will confirm interaction between these proteins in the cell lines.Keywords: annexin A2, calcineurin, CFTR, chloride, monocytes, pro-inflammatory molecules, S100A10
Procedia PDF Downloads 2352041 Survey of Selected Pathogenic Bacteria in Chickens from Rural Households in Limpopo Province
Authors: M. Lizzy Madiwani, Ignatious Ncube, Evelyn Madoroba
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This study was designed to determine the distribution of pathogenic bacteria in household raised chickens and study their virulence and antibiotic profiles. For this purpose, 40 chickens were purchased from families in the Capricorn district and sacrificed for sampling. Tissues were cultured on different bacteriological media followed by biotyping using Matrix-assisted Laser Desorption Ionization-time of Flight (MALDI-TOF). Disk diffusion test was performed to determine the antibiotic susceptibility profiles of these bacteria. Out of a total of 160 tissue samples evaluated, E. coli and Salmonella were detected in these tissues. Furthermore, determination of the pathogenic E. coli and Salmonella strains at species level using primer sets that target selected genes of interest in the polymerase chain reaction (PCR) assay was employed. The invA gene, a confirmatory gene of Salmonella was detected in all the Salmonella isolates. The study revealed that there is a high distribution of Salmonella and pathogenic E. coli in these chickens. Therefore, further studies on identification at the species level are highly recommended to provide management and sanitation practices to lower this prevalence. The antimicrobial susceptibly data generated from this study can be a valuable reference to veterinarians for treating bacterial diseases in poultry.Keywords: antimicrobial, Escherichia coli, pathogens, Salmonella
Procedia PDF Downloads 1332040 Development of Entrepreneurship in Industry on the Basis of Regulation of Transnational Production Chains in the Russian Arctic
Authors: E. N. Vetrova, L.V. Lapochkina, N. V. Nikulina
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In the national economy, entrepreneurship plays the role of a buffer between economy and policy for it contributes to improving budget effectiveness and decreasing dependence of economy on the state. Entrepreneurship in industry makes it possible to increase the added value that is formed in production chains and to decrease dependence on import. Under the current circumstances, when sanctions are being imposed, this is especially relevant for Russia and for the realization of projects in the Russian Arctic. However, development of entrepreneurship in industry requires an enlightened state policy. The purpose of the research is elaboration of recommendations for improving economic effectiveness of the realization of the Arctic projects on the basis of conceptual proposals for the development of entrepreneurship in industry. The paper presents the studies of the extractive industry role in the Russian economy and proves its raw material character. The analysis of production chains in industry on the basis of the conception of the added value global chains demonstrated a low added value formed by Russian companies. The study of changes in the structure of economy based on systemic, statistical and comparative analyses revealed no positive changes in the structure of economy over the period under consideration. This is a manifestation of ineffectiveness of the Russian industrial policy in general and within the Arctic region in particular. The authors identified the problems information and implementation of the state industrial policy in the Arctic region and in the development of national entrepreneurship, analyzed the shortcomings of the current state policy in the sphere of the Russian industry. On the basis of the conducted studies, the authors formulated conceptual approaches to change the state policy in the Arctic. The basic idea of the authors is to substantiate the focus of the state regulation on the development of entrepreneurship in industry in the process of the Russian Arctic exploration. At the same time another problem is solved–that of the development of the manufacturing industry in the southern regions of the northwestern part of Russia. The criterion of effectiveness in this case is the economic effectiveness.Keywords: entrepreneurship in industry, global chains of the added value, government regulation, industrial policies, production chains in the arctic region, economic effectiveness
Procedia PDF Downloads 3872039 The Problem of Legal Regulation of Joint Physical Custody: The Polish Perspective
Authors: Katarzyna Kamińska
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The main purpose of the work is to present the results of the studies regarding joint physical custody in the Polish legal system. The issues addressed fit into the ongoing process of modernising family law regulations and their adaptation to changing social reality in Poland. The Polish legislator now faces a dilemma: whether to introduce into Polish law a developed substantive or procedural regulation of joint physical custody and then whether it should be considered a legal presumption. Joint physical custody after divorce or separation is theoretically possible in Poland. It can either follow from the court’s independent proposal based on the assessment of the circumstances or from the parenting plan submitted by parents wishing to jointly retain full parental authority. However, joint physical custody does not result directly from the Polish Family and Guardianship Code. Therefore, there is real legal uncertainty in this matter, which leads to different treatment of citizens by the public authorities and courts. Another problem is that joint physical custody is misunderstood by the Polish courts. The main thesis of the work is that joint physical custody does not only mean the system of symmetrical child care (50/50), and the possibility to award joint physical custody will require the courts to carefully weigh the pros and cons of such an arrangement in each individual case.Keywords: joint physical custody, shared parenting, divorce, separation, parental authority
Procedia PDF Downloads 912038 SOCS3 Reverses Multidrug Resistance by Inhibiting MDR1 in Mammary Cell Carcinoma
Authors: S. Pradhan, D. Pradhan, G. Tripathy, T. Dasmohapatra
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Suppressors of cytokine signalling (SOCS3), a newly indentified anti-apoptotic molecule is a downstream effecter of the receptor tyrosine kinase-Ras signalling pathway. Current study has uncovered that SOCS3 may have wide and imperative capacities, particularly because of its close correlation with malignant tumors. To investigate the impact of SOCS3 on MDR, we analyzed the expression of P-gp and SOCS3 by immune-histochemistry and found there was positive correlation between them. At that point we effectively interfered with RNA translation by the contamination of siRNA of SOCS3 into MCF7/ADM breast cancer cell lines through a lentivirus, and the expression of the target gene was significantly inhibited. After RNAi the drug resistance was reduced altogether and the expression of MDR1 mRNA and P-gp in MCF7/ADM cell lines demonstrated a significant decrease. Likewise the expression of P53 protein increased in a statistically significant manner (p ≤ 0.01) after RNAi exposure. Moreover, flowcytometry analysis uncovers that cell cycle and anti-apoptotic enhancing capacity of cells changed after RNAi treatment. These outcomes proposed SOCS3 may take part in breast cancer MDR by managing MDR1 and P53 expression, changing cell cycle and enhancing the anti-apoptotic ability.Keywords: SOCS3gene, breast cancer, multidrug resistance, MDR1 gene, RNA interference
Procedia PDF Downloads 3402037 Molecular Characterization and Phylogenetic Analysis of Influenza a(H3N2) Virus Circulating during the 2010-2011 in Riyadh, Saudi Arabia
Authors: Ghazanfar Ali, Fahad N Almajhdi
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This study provides data on the viral diagnosis and molecular epidemiology of influenza A(H3N2) virus isolated in Riyadh, Saudi Arabia. Nasopharyngeal aspirates from 80 clinically infected patients in the peak of the 2010-2011 winter seasons were processed for viral diagnosis by RT-PCR. Sequencing of entire HA and NA genes of representative isolates and molecular epidemiological analysis were performed. A total of 06 patients were positive for influenza A, B and respiratory syncytial viruses by RT-PCR assays; out of these only one sample was positive for influenza A(H3N2) by RT-PCR. Phylogenetic analysis of the HA and NA gene sequences showed identities higher than 99-98.8 % in both genes. They were also similar to reference isolates in HA sequences (99 % identity) and in NA sequences (99 % identity). Amino acid sequences predicted for the HA gene were highly identical to reference strains. The NA amino acid substitutions identified did not include the oseltamivir-resistant H275Y substitution. Conclusion: Viral isolation and RT-PCR together were useful for diagnosis of the influenza A (H3N2) virus. Variations in HA and NA sequences are similar to those identified in worldwide reference isolates and no drug resistance was found.Keywords: influenza A (H3N2), genetic characterization, viral isolation, RT-PCR, Saudi Arabia
Procedia PDF Downloads 2642036 LncRNA NEAT1 Promotes NSCLC Progression through Acting as a ceRNA of miR-377-3p
Authors: Chengcao Sun, Shujun Li, Cuili Yang, Yongyong Xi, Liang Wang, Feng Zhang, Dejia Li
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Recently, the long non-coding RNA (lncRNA) NEAT1 has been identified as an oncogenic gene in multiple cancer types and elevated expression of NEAT1 was tightly linked to tumorigenesis and cancer progression. However, the molecular basis for this observation has not been characterized in progression of non-small cell lung cancer (NSCLC). In our studies, we identified NEAT1 was highly expressed in NSCLC patients and was a novel regulator of NSCLC progression. Patients whose tumors had high NEAT1 expression had a shorter overall survival than patients whose tumors had low NEAT1 expression. Further, NEAT1 significantly accelerates NSCLC cell growth and metastasis in vitro and tumor growth in vivo. Additionally, by using bioinformatics study and RNA pull down combined with luciferase reporter assays, we demonstrated that NEAT1 functioned as a competing endogenous RNA (ceRNA) for has-miR-377-3p, antagonized its functions and led to the de-repression of its endogenous targets E2F3, which was a core oncogene in promoting NSCLC progression. Taken together, these observations imply that the NEAT1 modulated the expression of E2F3 gene by acting as a competing endogenous RNA, which may build up the missing link between the regulatory miRNA network and NSCLC progression.Keywords: long non-coding RNA NEAT1, hsa-miRNA-377-3p, E2F3, non-small cell lung cancer, tumorigenesis
Procedia PDF Downloads 3752035 Mechanistic Structural Insights into the UV Induced Apoptosis via Bcl-2 proteins
Authors: Akash Bera, Suraj Singh, Jacinta Dsouza, Ramakrishna V. Hosur, Pushpa Mishra
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Ultraviolet C (UVC) radiation induces apoptosis in mammalian cells and it is suggested that the mechanism by which this occurs is the mitochondrial pathway of apoptosis through the release of cytochrome c from the mitochondria into the cytosol. The Bcl-2 family of proteins pro-and anti-apoptotic is the regulators of the mitochondrial pathway of apoptosis. Upon UVC irradiation, the proliferation of apoptosis is enhanced through the downregulation of the anti-apoptotic protein Bcl-xl and up-regulation of Bax. Although the participation of the Bcl-2 family of proteins in apoptosis appears responsive to UVC radiation, to the author's best knowledge, it is unknown how the structure and, effectively, the function of these proteins are directly impacted by UVC exposure. In this background, we present here a structural rationale for the effect of UVC irradiation in restoring apoptosis using two of the relevant proteins, namely, Bid-FL and Bcl-xl ΔC, whose solution structures have been reported previously. Using a variety of biophysical tools such as circular dichroism, fluorescence and NMR spectroscopy, we show that following UVC irradiation, the structures of Bcl-xlΔC and Bid-FL are irreversibly altered. Bcl-xLΔC is found to be more sensitive to UV exposure than Bid-FL. From the NMR data, dramatic structural perturbations (α-helix to β-sheet) are seen to occur in the BH3 binding region, a crucial segment of Bcl-xlΔC which impacts the efficacy of its interactions with pro-apoptotic tBid. These results explain the regulation of apoptosis by UVC irradiation. Our results on irradiation dosage dependence of the structural changes have therapeutic potential for the treatment of cancer.Keywords: Bid, Bcl-xl, UVC, apoptosis
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