Search results for: gene frequency

Authors: Marta E. Hernandez-Caballero, Veronica Borgonio-Cuadra, Antonio Miranda-Duarte, Xochitl Rojas-Toledo, Normand Garcia-Hernandez, Maura Cardenas-Garcia, Teresa Abad-Camacho

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Breast cancer (BC) is the most common tumor in women over the world. DNA methylation is an epigenetic modification critical in CpG sites, aberrant methylation of CpG islands in promoters is a hallmark of cancer. So, gene expression can be regulated by alterations in DNA methylation. In cell lines DACT2 gene reduces the growth and migration of tumor cells by its participation in the suppression of TGFb/SMAD2/3. PHF20L1 is involved in histone acetylation therefore, it regulates transcription. Our aim was to analyze the methylation status of the DACT2 and PHF20L1 promoter regions in tumoral and healthy mammary tissue from women with BC in different progression states. The study included 77 patients from Centro Medico Nacional La Raza in Mexico City. After identifying a CpG island in DACT2 and PHF20L1 promoters, DNA methylation status was analyzed through sodium bisulfite with subsequent amplification using methylation-specific PCR. Results revealed no changes in methylation status of PHF20L1 and cancer stages (II y III) or in comparison to healthy tissues, it was demethylated. DACT2 promoter methylation was no significant between tumoral stages (II, P = 0.37; III, P = 0.17) or with healthy tissue. Previous data reported DACT2 methylated in nasopharyngeal carcinoma but in this study promoter methylation was not observed. PHF20L1 protein contains N-terminal Tudor and C-terminal plant homeodomain domains, it has been suggested that can stabilize DNMT1 regulating DNA methylation, therefore, was associated with poor prognostic in BC. We found no evidence of methylation in patients and controls in PHF20L1 promoter, so its association with BC may have no direct relation with promoter methylation. More studies including other methylation sites in these genes in BC are necessary.

Keywords: bisulfite conversion, breast cancer, DACT2, DNA methylation, PHF20L1, tumoral status

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Authors: Anyimc, C. J. Aneke, J. O. Orji, O. Nworie, U. C. C. Egbule

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The microbial examination and antimicrobial susceptibility profile of microorganism isolated from the salt mining site in Ebonyi state were evaluated in the present study using a standard microbiological technique. A total of 300 samples were randomly collected in three sample groups (A, B, and C) of 100 each. Isolation, Identification and characterization of organization present on the soil samples were determined by culturing, gram-staining and biochemical technique. The result showed the following organisms were isolated with their frequency as follow: Bacillus species (37.3%) and Staphylococcus species(23.5%) had the highest frequency in the whole Sample group A and B while Klebsiella specie (15.7%), Pseudomonas species(13.7%), and Erwinia species (9.8%) had the least. Rhizopus species (42.0%) and Aspergillus species (26.0%) were the highest fungi isolated, followed by Penicillum species (20.0%) while Mucor species (4.0%), and Fusarium species (8.0%) recorded the least. Sample group C showed high microbial population of all the microbial isolates when compared to sample group A and B. Disc diffusion method was used to determine the susceptibility of isolated bacteria to various antibiotics (oxfloxacin, pefloxacin, ciprorex, augumentin, gentamycin, ciproflox, septrin, ampicillin), while agar well diffusion method was used to determine the susceptibility of isolated fungi to some antifungal drugs (metronidazole, ketoconazole, itraconazole fluconazole). The antibacterial activity of the antibiotics used showed that ciproflux has the best inhibitory effect on all the test bacteria. Ketoconazole showed the highest inhibitory effect on the fungal isolates, followed by itraconazole, while metronidazole and fluconazole showed the least inhibitory effect on the entire test fungal isolates. Hence, the multiple drug resistance of most isolates to appropriate drugs of choice are of great public health concern and cells for periodic monitoring of antibiograms to detect possible changing patterns. Microbes isolated in the salt mining site can also be used as a source of gene(s) that can increase salt tolerance in different crop species through genetic engineering.

Keywords: microorganisms, antibacterial, antifungal, resistance, salt mining site, Ebonyi State

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Authors: Chengcao Sun, Shujun Li, Cuili Yang, Yongyong Xi, Liang Wang, Feng Zhang, Dejia Li

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Hsa-miRNA-139-5p (miR-139-5p) has recently been discovered having anticancer efficacy in different organs. However, the role of miR-139-5p on lung cancer is still ambiguous. In this study, we investigated the role of miR-139-5p on development of lung cancer. Results indicated miR-139-5p was significantly down-regulated in primary tumor tissues and very low levels were found in a non-small cell lung cancer (NSCLC) cell lines. Ectopic expression of miR-139-5p in NSCLC cell lines significantly suppressed cell growth through inhibition of cyclin D1 and up-regulation of p57(Kip2). In addition, miR-139-5p induced apoptosis, as indicated by up-regulation of key apoptosis gene cleaved caspase-3, and down-regulation of anti-apoptosis gene Bcl2. Moreover, miR-139-5p inhibited cellular metastasis through inhibition of matrix metalloproteinases (MMP)-7 and MMP-9. Further, oncogene c-Met was revealed to be a putative target of miR-139-5p, which was inversely correlated with miR-139-5p expression. Taken together, our results demonstrated that miR-139-5p plays a pivotal role in lung cancer through inhibiting cell proliferation, metastasis, and promoting apoptosis by targeting oncogenic c-Met.

Keywords: hsa-miRNA-139-5p (miR-139-5p), c-Met, non-small cell lung cancer (NSCLC), proliferation, apoptosis

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Authors: Arti Devi, Parthasarthi Choudhury

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Extreme rainfall frequency analysis for Meteorological Sub-Division 4 of India was analysed using L-moments approach. Serial Correlation and Mann Kendall tests were conducted for checking serially independent and stationarity of the observations. The discordancy measure for the sites was conducted to detect the discordant sites. The regional homogeneity was tested by comparing with 500 generated homogeneous regions using a 4 parameter Kappa distribution. The best fit distribution was selected based on ZDIST statistics and L-moments ratio diagram from the five extreme value distributions GPD, GLO, GEV, P3 and LP3. The LN3 distribution was selected and regional rainfall frequency relationship was established using index-rainfall procedure. A regional mean rainfall relationship was developed using multiple linear regression with latitude and longitude of the sites as variables.

Keywords: L-moments, ZDIST statistics, serial correlation, Mann Kendall test

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Authors: Y. Hu, X. Zhao, T. Yamaguchi, M. Sasajima, Y. Koike

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To focus on the vibration mode of a cone loudspeaker, which acts as an electroacoustic transducer, excitation experiments were performed using two types of loudspeaker units: one employing an impulse hammer and the other a sweep signal. The on-axis sound pressure frequency properties of the loudspeaker were evaluated, and the characteristic properties of the loudspeakers were successfully determined in both excitation experiments. Moreover, under conditions identical to the experiment conditions, a coupled analysis of the vibration-acoustics of the cone loudspeaker was performed using an acoustic analysis software program that considers the impact of damping caused by air viscosity. The result of sound pressure frequency properties with the numerical analysis are the most closely match that measured in the excitation experiments over a wide range of frequency bands.

Keywords: anechoic room, finite element method, impulse hammer, loudspeaker, reverberation room, sweep signal

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Authors: Herlina Abdul Rahim, Javad Abbaszadeh, Ruzairi Abdul Rahim

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In this research, the non-invasive ultrasonic transmission tomography is investigated. In order to model the ultrasonic wave scattering for different thickness of metal pipes, two-dimensional (2D) finite element modeling (FEM) has been utilized. The wall thickness variation of the metal pipe and its influence on propagation of the ultrasonic pressure wave are explored in this paper, includes frequency analysing in order to find the maximum applicable frequency. The simulation results have been compared to experimental data and are shown to provide key insight for this well-defined experimental case by explaining the achieved reconstructed images from experimental setup. Finally, the experimental results which are useful for further investigation for the application of ultrasonic transmission tomography in industry are illustrated.

Keywords: ultrasonic transmission tomography, ultrasonic sensors, ultrasonic wave, non-invasive tomography, metal pipe

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Authors: Jin Young Kim, Kwon Kyoo Kang

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The SGR1 (STAYGREEN1) protein is a critical regulator of plant leaves in chlorophyll degradation and senescence. The functions and mechanisms of tomato SGR1 action are poorly understood and worthy of further investigation. To investigate the function of the SGR1 gene, we generated a SGR1-knockout (KO) null line via clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9-mediated gene editing and conducted RNA sequencing and gas chromatography tandem mass spectrometry (GC-MS/MS) analysis to identify the differentially expressed genes. The SlSGR1 (Solanum lycopersicum SGR1) knockout null line clearly showed a turbid brown color with significantly higher chlorophyll and carotenoid content compared to wild-type (WT) fruit. Differential gene expression analysis revealed 728 differentially expressed genes (DEGs) between WT and sgr1 #1-6 line, including 263 and 465 downregulated and upregulated genes, respectively, for which fold change was >2, and the adjusted p-value was <0.05. Most of the DEGs were related to photosynthesis and chloroplast function. In addition, the pigment, carotenoid changes in sgr1 #1-6 line was accumulated of key primary metabolites such as sucrose and its derivatives (fructose, galactinol, raffinose), glycolytic intermediates (glucose, G6P, Fru6P) and tricarboxylic acid cycle (TCA) intermediates (malate and fumarate). Taken together, the transcriptome and metabolite profiles of SGR1-KO lines presented here provide evidence for the mechanisms underlying the effects of SGR1 and molecular pathways involved in chlorophyll degradation and carotenoid biosynthesis.

Keywords: tomato, CRISPR/Cas9, null line, RNA-sequencing, metabolite profiling

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Authors: Omaima A. Sharaf, Wafaa M. Abd El-Rahim, Hassan Moawad, Michael J. Sadowsky

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Textile industry is one of the important industries in the worldwide. It is known that the eco-friendly industrial and agricultural activities are significant for socio-economic stability of all countries. The absence of appropriate industrial waste water treatments is essential barrier for sustainable development in food and agricultural sectors especially in developing country like Egypt. Thus, the development of enzymatic bioremediation technology for textile dye removal will enhance the collaboration between scientists who develop the technology and industry where this technology will be implemented towards the safe disposal of the textile dye wastes. Highly efficient microorganisms are of most importance in developing and using highly effective biological treatment processes. Bacterial degradation of azo dyes is generally initiated by an enzymatic step that involves cleavage of azo linkages, usually with the aid of an azoreductase as electron donor. Thus, expanding the spectrum of microorganisms with high enzymatic activities as azoreductases and discovering novel azo-dye degrading enzymes, with enhanced stability and superior catalytic properties, are necessary for many environmental and industrial applications. Consequently, the use of molecular tools has become increasingly integrated into the understanding of enzyme properties and characterization. Researchers have utilized a gene cloning and expression methods as a tool to produce recombinant protein for decolorizing dyes more efficiently. Thus, presumptive evidence for the presence of genes encoding azoreductases in the genomes of selected local, and most potent azo-degrading strains were obtained by using specific oligonucleotides primers. These potent strains have been isolated from textile industrial wastewater in Egypt and identified using 16S rRNA sequence analysis as 'Lysinibacillus macrolidesB8, Brevibacillus parabrevisB11, Bacillus coagulansB7, and B. cereusB5'. PCR products of two full length genes designated as (AZO1;621bp and AZO2;534bp) were detected. BLASTx results indicated that AZO1 gene was corresponding to predicted azoreductase from of Bacillus sp. ABP14, complete genome, multispecies azoreductase [Bacillus], It was submitted to the gene bank by an accession no., BankIt2085371 AZO1 MG923210 (621bp; 207 amino acids). AZO1 was generated from the DNA of our identified strains Lysinibacillus macrolidesB8. On the other hand, AZO2 gene was corresponding to a predicted azoreductase from Bacillus cereus strain S2-8. Gene bank accession no. was BankIt2085839 AZO2 MG932081 (534bp;178 amino acids) and it was amplified from our Bacillus coagulansB7. Both genes were successfully cloned into pCR2.1TOPO (Invitrogen) and in pET28b+ vectors, then they transformed into E. coli DH5α and BL21(DE3) cells for heterologous expression studies. Our recombinant azoreductases (AZO1&AZO2) exhibited potential enzyme activity and efficiently decolorized an azo dye (Direct violet). They exhibited pH stability between 6 and 8 with optimum temperature up to 60°C and 37 °C after induction by 1mM and 1.5mM IPTG, for both AZO1 &AZO2, respectively. These results suggested that further optimization and purification of these recombinant proteins by using different heterologous expression systems will give great potential for the sustainable utilization of these recombinant enzymes in several industrial applications especially in wastewater treatments.

Keywords: azoreductases, decolorization, enzyme activity, gene cloning and expression

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Authors: Erin Louise Robinson, Emily Elsa Freeman

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Rough-and-tumble play (RTP) is a physical and competitive type of play that parents engage in with their children. While past research has reported RTP to be the preferred play type for western fathers, the frequency of these interactions in Australian families have not been explored. With parental perceptions of play importance playing a major role in the frequency of activity engagement, the present study investigated how perceptions and parent gender impact on RTP play frequency. By utilising child gender in our approach, we also examined the historical trend of boys receiving more physical play interactions with their parents. Three hundred and seventy-nine respondents completed the study with their 0–10-year-old children. The results indicated that, in line with past research, parents engaged more frequently in RTP with their sons than their daughters. While, both mothers and fathers participated in RTP with their children, fathers perceived RTP to be of greater important to their child’s development than mothers did. Moreover, supporting previous findings, this more positive perception of the play was related to greater frequency of RTP in these father-child dyads. Although RTP literature remains heavily focussed on fathers, the fact that mothers are engaging in these interactions as well, establishes the need to explore maternal influences in future research.

Keywords: parenting, play, child development, family, Australia

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Authors: Redvan Ghasemlounia, Elham Ansari, Hikmet Kerem Cigizoglu

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Hydrological modeling in arid and semi-arid regions is very important. Iran has many regions with these climate conditions such as Chaharmahal and Bakhtiari province that needs lots of attention with an appropriate management. Forecasting of hydrological parameters and estimation of hydrological events of catchments, provide important information that used for design, management and operation of water resources such as river systems, and dams, widely. Discharge in rivers is one of these parameters. This study presents the application and comparison of some estimation methods such as Feed-Forward Back Propagation Neural Network (FFBPNN), Multi Linear Regression (MLR), Gene Expression Programming (GEP) and Bayesian Network (BN) to predict the daily flow discharge of the Soolegan River, located at Chaharmahal and Bakhtiari province, in Iran. In this study, Soolegan, station was considered. This Station is located in Soolegan River at 51° 14՜ Latitude 31° 38՜ longitude at North Karoon basin. The Soolegan station is 2086 meters higher than sea level. The data used in this study are daily discharge and daily precipitation of Soolegan station. Feed Forward Back Propagation Neural Network(FFBPNN), Multi Linear Regression (MLR), Gene Expression Programming (GEP) and Bayesian Network (BN) models were developed using the same input parameters for Soolegan's daily discharge estimation. The results of estimation models were compared with observed discharge values to evaluate performance of the developed models. Results of all methods were compared and shown in tables and charts.

Keywords: ANN, multi linear regression, Bayesian network, forecasting, discharge, gene expression programming

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Authors: Himanshu J. Bahirat, Apoorva S. Janawlekar

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Plasma actuators are essential for aerodynamic flow separation control due to their lack of mechanical parts, lightweight, and high response frequency, which have numerous applications in hypersonic or supersonic aircraft. The working of these actuators is based on the formation of a low-temperature plasma between a pair of parallel electrodes by the application of a high-voltage AC signal across the electrodes, after which air molecules from the air surrounding the electrodes are ionized and accelerated through the electric field. The high-frequency operation is required in dielectric discharge barriers to ensure plasma stability. To carry out flow separation control in a hypersonic flow, the optimal design and construction of a power supply to generate dielectric barrier discharges is carried out in this paper. In this paper, it is aspired to construct a simplified circuit topology to emulate the dielectric barrier discharge and study its various frequency responses. The power supply can generate high voltage pulses up to 20kV at the repetitive frequency range of 20-50kHz with an input power of 500W. The power supply has been designed to be short circuit proof and can endure variable plasma load conditions. Its general outline is to charge a capacitor through a half-bridge converter and then later discharge it through a step-up transformer at a high frequency in order to generate high voltage pulses. After simulating the circuit, the PCB design and, eventually, lab tests are carried out to study its effectiveness in controlling flow separation.

Keywords: aircraft propulsion, dielectric barrier discharge, flow separation control, power source

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Authors: Naso Isaiah Thanavisuth

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Introduction: Medical cannabis can be used for treatment, including anorexia, pain, inflammation, multiple sclerosis, Parkinson's disease, epilepsy, cancer, and metabolic syndrome-related disorders. However, medical cannabis leads to adverse effects (AEs), which is delta-9-tetrahydrocannabinol (THC). In previous studies, the major of THC metabolism enzymes are CYP2C9. Especially, the variation of CYP2C9 gene consist of CYP2C9*2 on exon 3 (C430T) (Arg144Cys) and CYP2C9*3 on exon 7 (A1075C) (Ile359Leu) to decrease enzyme activity. Notwithstanding, there is no data describing whether the variant of CYP2C9 genes are a pharmacogenetics marker for prediction of THC-induced AEs in Thai patients. Objective: We want to investigate the association between CYP2C9 gene and THC-induced AEs in Thai patients. Method: We enrolled 39 Thai patients with medical cannabis treatment consisting of men and women who were classified by clinical data. The quality of DNA extraction was assessed by using NanoDrop ND-1000. The CYP2C9*2 and *3 genotyping were conducted using the TaqMan real time PCR assay (ABI, Foster City, CA, USA). Results: All Thai patients who received the medical cannabis consist of twenty four (61.54%) patients who were female and fifteen (38.46%) were male, with age range 27- 87 years. Moreover, the most AEs in Thai patients who were treated with medical cannabis between cases and controls were tachycardia, arrhythmia, dry mouth, and nausea. Particularly, thirteen (72.22%) medical cannabis-induced AEs were female and age range 33 – 69 years. In this study, none of the medical cannabis groups carried CYP2C9*2 variants in Thai patients. The CYP2C9*3 variants (*1/*3, intermediate metabolizer, IM) and (*3/*3, poor metabolizer, PM) were found, three of thirty nine (7.69%) and one of thirty nine (2.56%) , respectively. Conclusion: This is the first study to confirm the genetic polymorphism of CYP2C9 and medical cannabis-induced AEs in the Thai population. Although, our results indicates that there is no found the CYP2C9*2. However, the variation of CYP2C9 allele might serve as a pharmacogenetics marker for screening before initiating the therapy with medical cannabis for prevention of medical cannabis-induced AEs.

Keywords: CYP2C9, medical cannabis, adverse effects, THC, P450

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Authors: Md. Shafiullah, M. Shafiul Alam, Bandar Suliman Alsharif

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Electrical quantities such as frequency, voltage, current are being fluctuated due to abnormalities in power system. Most of the abnormalities cause fluctuation in system frequency and sometimes extreme abnormalities lead to system blackout. To protect the system from complete blackout planned and proper islanding plays a very important role even in case of extreme abnormalities. Islanding operation not only helps stabilizing a faulted system but also supports power supplies to critical and important loads, in extreme emergency. But the islanding systems are weaker than integrated system so the stability of islands is the prime concern when an integrated system is disintegrated. In this paper, different impacts on a load rich island have been studied and a frequency based auto-load shedding scheme has been developed for sudden load addition, generation outage and combined effect of both to the island. The developed scheme has been applied to Khulna-Barisal Island to validate the effectiveness of the developed technique. Various types of abnormalities to the test system have been simulated and for the simulation purpose CYME PSAF (Power System Analysis Framework) has been used.

Keywords: auto load shedding, FS&FD relay, impact study, island, PSAF, ROCOF

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Authors: Vidya Padmakumar, N. C. Tharavathy

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The study was conducted from July 2015 to July 2017 to determine and understand the occurrence, frequency and diversity of avifauna in the Mallathahalli Lake of Bangalore Urban district. During the study period, 46 species of both terrestrial, as well as, aquatic birds belonging to 30 families were identified out of which 9 families were aquatic birds and 21 families were terrestrial birds. There were 4 species of migratory birds out of 46, showing diurnal migration. There was a significant reduce in the number of bird species both terrestrial and aquatic during the summer season and also varied greatly during winters and monsoon. Of the total 24 species of aquatic birds, Fulica atra and Tachybaptus ruficolis were the most common with 100% frequency and the least frequent species with 3.02% frequency was identified as Threskiornis melanocephalus. Among the 22 species of terrestrial birds, Acridotheres tristis had a frequency of 89% and the least frequent was Pycnonotus cafer (4.45%). The most commonly encountered bird species were from the families- Anatidae, Podicipedidae, Ardeidae, Phalacrocoracidae, Rallidae, Accipitridae, Scolopacidae, Charadridae, Laridae, Meropidae, Hirudinidae. All the birds surviving around the area are dependent on the wetland and crop vegetation surrounding the lake, which are deteriorating due to anthropogenic interventions and urbanization which are rising to its peak gradually causing the decline in the avifaunal diversity.

Keywords: Avifaunal diversity, Mallathahalli lake, seasonal migration, urbanization

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Authors: Zahra Solgi, Hossein Rassi

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Colorectal cancer (CRC) is a multi-step disease, and chronic gastric infection with H. pylori could play a role in one or more of the steps in this pathogenic process. Polymorphisms in several miRNAs are considered to increase the risk for the development of CRC by controlling proliferation, apoptosis and H. pylori pathogenesis. Therefore, the aim of this study was to investigate miRNA146a rs2910164 polymorphism and Helicobacter pylori infection in CRC. A total of 65 patients with CRC were divided into 2 groups: 28 patients < 50 years of age and 37 patients ≥ 50 years of age. DNA was extracted from all samples by a standard method and H. pylori cagA and miRNA146a rs2910164 genotypes were determined by PCR method. The results show that there was no significant difference in the frequency of H. pylori cagA gene between the two groups but there was a significant difference in the distribution of rs2910164 genotypes in patients < 50 years of age with the p-value of 0.05 and odds ratio equal to 2.69. On other hand, patients < 50 years of age with genotype CC of miRNA146a showed a significant difference in CRC risk. Furthermore, there was a significant correlation between rs2910164 CC genotype with Helicobacter pylori infection in patients < 50 years of age. The present study suggests that the CC genotype of miRNA146a in combination with H. pylori infection can be effective as risk factors and molecular markers for early diagnosis and treatment of CRC.

Keywords: colorectal cancer, Helicobacter pylori, miRNA146a, rs2910164 polymorphism

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Authors: Atsuko Okazaki, Jurg Ott

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Some genetic diseases (‘digenic traits’) are due to the interaction between two DNA variants. For example, certain forms of Retinitis Pigmentosa (a genetic form of blindness) occur in the presence of two mutant variants, one in the ROM1 gene and one in the RDS gene, while the occurrence of only one of these mutant variants leads to a completely normal phenotype. Detecting such digenic traits by genetic methods is difficult. A common approach to finding disease-causing variants is to compare 100,000s of variants between individuals with a trait (cases) and those without the trait (controls). Such genome-wide association studies (GWASs) have been very successful but hinge on genetic effects of single variants, that is, there should be a difference in allele or genotype frequencies between cases and controls at a disease-causing variant. Frequent pattern mining (FPM) methods offer an avenue at detecting digenic traits even in the absence of single-variant effects. The idea is to enumerate pairs of genotypes (genotype patterns) with each of the two genotypes originating from different variants that may be located at very different genomic positions. What is needed is for genotype patterns to be significantly more common in cases than in controls. Let Y = 2 refer to cases and Y = 1 to controls, with X denoting a specific genotype pattern. We are seeking association rules, ‘X → Y’, with high confidence, P(Y = 2|X), significantly higher than the proportion of cases, P(Y = 2) in the study. Clearly, generally available FPM methods are very suitable for detecting disease-associated genotype patterns. We use fpgrowth as the basic FPM algorithm and built a framework around it to enumerate high-frequency digenic genotype patterns and to evaluate their statistical significance by permutation analysis. Application to a published dataset on opioid dependence furnished results that could not be found with classical GWAS methodology. There were 143 cases and 153 healthy controls, each genotyped for 82 variants in eight genes of the opioid system. The aim was to find out whether any of these variants were disease-associated. The single-variant analysis did not lead to significant results. Application of our FPM implementation resulted in one significant (p < 0.01) genotype pattern with both genotypes in the pattern being heterozygous and originating from two variants on different chromosomes. This pattern occurred in 14 cases and none of the controls. Thus, the pattern seems quite specific to this form of substance abuse and is also rather predictive of disease. An algorithm called Multifactor Dimension Reduction (MDR) was developed some 20 years ago and has been in use in human genetics ever since. This and our algorithms share some similar properties, but they are also very different in other respects. The main difference seems to be that our algorithm focuses on patterns of genotypes while the main object of inference in MDR is the 3 × 3 table of genotypes at two variants.

Keywords: digenic traits, DNA variants, epistasis, statistical genetics

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Authors: Hui Li, Xueying Zhao, Ke Ma, Yu Cao, Fan Yang, Qingwen Xu, Wenbin Liu

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Soil can often link a person or item to a crime scene, which makes it a valuable evidence in forensic casework. Several techniques have been utilized in forensic soil discrimination in previous studies. Because soil contains a vast number of microbiomes, the analyse of soil microbiomes is expected to be a potential way to characterise soil evidence. In this study, we applied massively parallel sequencing (MPS) to soil bacterial profiling on the Ion Torrent Personal Genome Machine (PGM). Soils from different regions were collected repeatedly. V-region 3 and 4 of Bacterial 16S rRNA gene were detected by MPS. Operational taxonomic units (OTU, 97%) were used to analyse soil bacteria. Several bioinformatics methods (PCoA, NMDS, Metastats, LEfse, and Heatmap) were applied in bacterial profiles. Our results demonstrate that MPS can provide a more detailed picture of the soil microbiomes and the composition of soil bacterial components from different region was individualistic. In conclusion, the utility of soil bacterial profiling via MPS of the 16S rRNA gene has potential value in characterising soil evidences and associating them with their place of origin, which can play an important role in forensic science in the future.

Keywords: bacterial profiling, forensic, massively parallel sequencing, soil evidence

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Authors: Mohammad Naeem Lakanwall, Jamshid Abdul-Ghafar

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Background: Endocrinopathies are a commonly occurring entity, particularly those of the thyroid gland; however, there is a lack of scientific literature from Afghanistan, a country with very limited health care facilities and resources. To our best knowledge, this is the first study aimed to describe the frequency of occurrence and factors associated with thyroid dysfunction in the Afghan population. The aim of this study is to estimate the frequency and to identify factors associated with thyroid dysfunction among individuals coming to a tertiary care facility in Kabul, Afghanistan. Methods: A cross-sectional study was conducted from July to Sep 2018 at the Department of Clinical Pathology, French Medical Institute for Mothers and Children (FMIC), Kabul, Afghanistan. Blood samples were obtained, serum TSH levels were analyzed, and the patients were divided into three diagnostic categories according to their serum TSH concentrations: 1) hypothyroidism, 2) hyperthyroidism, 3) normal. Results: A total of 127 individuals were included in the final analysis. The majority of study participants (77%) were females. A large number of the participants (92%) did not have a family history of thyroid dysfunction. 74% of the participants in the study had normal TSH levels classified as normal thyroid function, (14%) had lower TSH levels, and (12%) higher TSH levels, classified as hyper and hypothyroid, respectively. Conclusions: The findings of the current study showed a high frequency of thyroid dysfunctions from a single center. Further large-scale studies are needed to find out the prevalence and document this entity for better health outcomes in the country.

Keywords: Afghanistan, factors, frequency, hypothyroid, hyperthyroid, thyroid, thyroid stimulating hormone

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Authors: Yusuf Ulaş Kabukçu, Si̇nan Çeli̇k, Onur Salan, Mai̇de Altuntaş, Mert Can Dalkiran, Gökseni̇n Bozdağ, Metehan Bulut, Fati̇h Yaman

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This study presents a more compact implementation of the 2.4GHz MIT Coffee Can Doppler Radar for 2.6GHz operating frequency. The main difference of our prototype depends on the use of microstrip antennas which makes it possible to transport with a small robotic vehicle. We have designed our radar system with two different channels: Tx and Rx. The system mainly consists of Voltage Controlled Oscillator (VCO) source, low noise amplifiers, microstrip antennas, splitter, mixer, low pass filter, and necessary RF connectors with cables. The two microstrip antennas, one is element for transmitter and the other one is array for receiver channel, was designed, fabricated and verified by experiments. The system has two operation modes: speed detection and range detection. If the switch of the operation mode is ‘Off’, only CW signal transmitted for speed measurement. When the switch is ‘On’, CW is frequency-modulated and range detection is possible. In speed detection mode, high frequency (2.6 GHz) is generated by a VCO, and then amplified to reach a reasonable level of transmit power. Before transmitting the amplified signal through a microstrip patch antenna, a splitter used in order to compare the frequencies of transmitted and received signals. Half of amplified signal (LO) is forwarded to a mixer, which helps us to compare the frequencies of transmitted and received (RF) and has the IF output, or in other words information of Doppler frequency. Then, IF output is filtered and amplified to process the signal digitally. Filtered and amplified signal showing Doppler frequency is used as an input of audio input of a computer. After getting this data Doppler frequency is shown as a speed change on a figure via Matlab script. According to experimental field measurements the accuracy of speed measurement is approximately %90. In range detection mode, a chirp signal is used to form a FM chirp. This FM chirp helps to determine the range of the target since only Doppler frequency measured with CW is not enough for range detection. Such a FMCW Doppler radar may be used in border security of the countries since it is capable of both speed and range detection.

Keywords: doppler radar, FMCW, range detection, speed detection

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Authors: S. Rostampour Yasouri, M. Ghane

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Leptospirosis is one the most important common diseases between human and live stock occurred by different species of Leptospira. This disease has been construed as the native in the northern provinces of Iran and risk of the infection with pathogenic is high. One hundred fifteen samples of water (67), soil (36) and feces of rodents (12) were collected from the rice fields of the suburbs of Tonekabon Township situated in northern part of Iran in 2012. The samples, after passage from membranous filters, were cultured in the liquid and solid EMJH medium and incubated at 30°C for 1 month. Leptospira spp. were isolated using culture technique, and the plates were studied from viewpoint of colony formation, microscopic observations and then identified by phenotyping tests. Finally, the identification of Leptospira genus was verified by PCR technique and 16S rRNA gene sequencing. Of 115 samples totally, 55 samples (47.82%) became positive by use of the culture technique which the positive cases included 47 water samples (70.14%) and 8 soil samples (22.22%), while the isolation was not accomplished from the sample of the rodents feces. Overall, according to these data, Leptospira spp. exists with high frequency in North Iran. Hence, based on foregoing evidence environments in the north of Iran are vehicles of Leptospira spp.

Keywords: EMJH Medium, Leptospira, Northern of Iran, rice fields

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Authors: Ahmed A. D. Sarhan, S. R. Besharaty, Javad Akbaria, M. Hamdi

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The capability of CNC gantry milling machines in manufacturing long components has caused the expanded use of such machines. On the other hand, the machines’ gantry rigidity can reduce under severe loads or vibration during operation. Indeed, the quality of machining is dependent on the machine’s dynamic behavior throughout the operating process. For this reason, this type of machines has always been used prudently and are non efficient. Therefore, they can usually be employed for rough machining and may not produce adequate surface finishing. In this paper, a CNC gantry milling machine with the potential to produce good surface finish has been designed and analyzed. The lowest natural frequency of this machine is 202 Hz at all motion amplitudes with a full range of suitable frequency responses. Meanwhile, the maximum deformation under dead loads for the gantry machine is 0.565µm, indicating that this machine tool is capable of producing higher product quality.

Keywords: frequency response, finite element, gantry machine, gantry design, static and dynamic analysis

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Authors: David D. Adams, Ibrahim B. Bello

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Spent oil contaminated Soil from ten selected mechanic workshops were investigated for their bacteria and bioremediation potentials. The bacterial isolates were morphologically and molecularly identified as Enterobacter hormaechei, Escherichia coli, Klebsiella pneumoniae, Shigella flexneri , Wesiella cibaria, Lactobacillus planetarium. The singles and a consortium of these bacteria incubated in the minimal salt medium incorporated with 1% engine oil exhibited various biodegradation rates, with the mixed consortium exhibiting the highest for this oil. The gene for the hydrocarbon enzyme Catechol 2, 3 dioxygenase (C2,30) was detected and amplified in Enterobacter hormaechei, Escherichia coli and Shigella flexneri using PCR and Agarose gel electrophoresis. The detection of the (C2,30) enzyme gene in, and the spent oil biodegradation activity exhibited by these bacteria suggest their possible possession of bioremediating potentials for the spent engine oil. It is therefore suggested that a pilot study on the field application of these bacteria for bioremediation and restoration of spent oil polluted environment should be done in mechanic workshops.

Keywords: spent engine oil, pollution, bacteria, enzyme, bioremediation, mechanic workshop

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Authors: Abla A. Ali, Heba A. Abd El Razik, Nadia A. Kotb, Amany A. Bayoumi, Laila A. Rashed

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The detection of human skin through mRNA-based profiling is a very useful tool for forensic investigations. The aim of this study was definitive identification of human skin at different time intervals using an mRNA marker late cornified envelope gene 1C. Ten middle-aged healthy volunteers of both sexes were recruited for this study. Skin samples controlled with blood samples were taken from the candidates to test for the presence of our targeted mRNA marker. Samples were kept at dry dark conditions to be tested at different time intervals (24 hours, one week, three weeks and four weeks) for detection and relative quantification of the targeted marker by RT PCR. The targeted marker could not be detected in blood samples. The targeted marker showed the highest mean value after 24 hours (11.90 ± 2.42) and the lowest mean value (7.56 ± 2.56) after three weeks. No marker could be detected at four weeks. This study verified the high specificity and sensitivity of mRNA marker in the skin at different storage times up to three weeks under the study conditions.

Keywords: human skin, late cornified envelope gene 1C, mRNA marker, time intervals

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Authors: Qiuyi Lyu, Bin Gong

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Breadth-First Search(BFS) is a core graph algorithm that is widely used for graph analysis. As it is frequently used in many graph applications, improve the BFS performance is essential. In this paper, we present a graph ordering method that could reorder the graph nodes to achieve better data locality, thus, improving the BFS performance. Our method is based on an observation that the sibling relationships will dominate the cache access pattern during the BFS traversal. Therefore, we propose a frequency-based model to construct the graph order. First, we optimize the graph order according to the nodes’ visit frequency. Nodes with high visit frequency will be processed in priority. Second, we try to maximize the child nodes overlap layer by layer. As it is proved to be NP-hard, we propose a heuristic method that could greatly reduce the preprocessing overheads. We conduct extensive experiments on 16 real-world datasets. The result shows that our method could achieve comparable performance with the state-of-the-art methods while the graph ordering overheads are only about 1/15.

Keywords: breadth-first search, BFS, graph ordering, graph algorithm

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Authors: Arash Yousefi

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Experts suggest that relying exclusively on parameters to convey a situation or establish a condition may not be adequate. Assessing and appraising incidents in a system based on accident parameters, such as accident frequency, lost workdays, or fatalities, may not always be precise and occasionally erroneous. The frequency rate of accidents is a metric that assesses the correlation between the number of accidents causing work-time loss due to injuries and the total working hours of personnel over a year. Traditionally, this has been calculated based on one million working hours, but the American Occupational Safety and Health Organization has updated its standards. The new coefficient of 200/000 working hours is now used to compute the frequency rate of accidents. It's crucial to ensure that the total working hours of employees are equally represented when calculating individual event and incident numbers. The accident severity rate is a metric used to determine the amount of time lost or wasted during a given period, often a year, in relation to the total number of working hours. It measures the percentage of work hours lost or wasted compared to the total number of useful working hours, which provides valuable insight into the number of days lost or wasted due to work-related incidents for each working hour. Calculating the severity of an incident can be difficult if a worker suffers permanent disability or death. To determine lost days, coefficients specified in the "tables of days equivalent to OSHA or ANSI standards" for disabling injuries are used. The accident frequency coefficient denotes the rate at which accidents occur, while the accident severity coefficient specifies the extent of damage and injury caused by these accidents. These coefficients are crucial in accurately assessing the magnitude and impact of accidents.

Keywords: incidents, safety, analysis, frequency, severity, injuries, determine

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Authors: Sahar Hooshmand, Mohammad Reza Bagherzadeh Anasari

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The aim of this study was to examine the effect of breakfast eating pattern (between frequencies of breakfast consumers and non-consumers) on nutritional status (weight for age, height for age and weight for height). A total 4570 primary school children aged 6-9 years old constituted the sample. From these, 2234 Iranian school children (1218 girls and 1016 boys) and 2336 Indian school children (1096 girls and 1240 boys) were included in a cross sectional study. Breakfast frequency consumption was recorded through an interview with mothers of children. Height and wight of children were taken and body mass index were calculated. The World Health Organization’s (WHO) AnthroPlus software used to assess the nutritional status of the children. Weight for age z-scores were slightly associated with frequency of consuming breakfast in both India (χ2 = 60.083, p=0.000) and Iran (χ2 = 18.267, p=0.032). A significant association was seen between frequency of child‘s breakfast intake and the height z-scores in both India (χ2 = 31.334, p=0.000) and Iran (χ2 = 19.443, p=0.022). Most of children with normal height had breakfast daily in both countries. A significant association was seen with children‘s BMI z-scores of Indian children (χ2 = 31.247, p=0.000) but it was not significant in Iran (χ2 = 10.791, p=0.095). The present study confirms the observations of other studies that showed more frequency in having breakfast is associated with better nutritional status.

Keywords: breakfast, schoolchildren, nutritional status, global food security

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Authors: Hamzeh Alipour, Masoumeh Bagheri, Abbasali Raz, Javad Dadgar Pakdel, Kourosh Azizi, Aboozar Soltani, Mohammad Djaefar Moemenbellah-Fard

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Background: Maggot debridement therapy is an appropriate, effective, and controlled method using sterilized larvae of Luciliasericata (L.sericata) to treat wounds. Netrin-A is an enzyme in the Laminins family which secreted from salivary gland of L.sericata with a central role in neural regeneration and angiogenesis. This study aimed to production of new recombinant Netrin-A protein of Luciliasericata larvae by baculovirus expression vector system (BEVS) in SF9. Material and methods: In the first step, gene structure was subjected to the in silico studies, which were include determination of Antibacterial activity, Prion formation risk, homology modeling, Molecular docking analysis, and Optimization of recombinant protein. In the second step, the Netrin-A gene was cloned and amplified in pTG19 vector. After digestion with BamH1 and EcoR1 restriction enzymes, it was cloned in pFastBac HTA vector. It was then transformed into DH10Bac competent cells, and the recombinant Bacmid was subsequently transfected into insect Sf9 cells. The expressed recombinant Netrin-A was thus purified in the Ni-NTA agarose. This protein evaluation was done using SDS-PAGE and western blot, respectively. Finally, its concentration was calculated with the Bradford assay method. Results: The Bacmid vector structure with Netrin-A was successfully constructed and then expressed as Netrin-A protein in the Sf9 cell lane. The molecular weight of this protein was 52 kDa with 404 amino acids. In the in silico studies, fortunately, we predicted that recombinant LSNetrin-A have Antibacterial activity and without any prion formation risk.This molecule hasa high binding affinity to the Neogenin and a lower affinity to the DCC-specific receptors. Signal peptide located between amino acids 24 and 25. The concentration of Netrin-A recombinant protein was calculated to be 48.8 μg/ml. it was confirmed that the characterized gene in our previous study codes L. sericata Netrin-A enzyme. Conclusions: Successful generation of the recombinant Netrin-A, a secreted protein in L.sericata salivary glands, and because Luciliasericata larvae are used in larval therapy. Therefore, the findings of the present study could be useful to researchers in future studies on wound healing.

Keywords: blowfly, BEVS, gene, immature insect, recombinant protein, Sf9

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Authors: Ahmet Kayabasi, Ali Akdagli

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In this study, three robust predicting methods, namely artificial neural network (ANN), adaptive neuro fuzzy inference system (ANFIS) and support vector machine (SVM) were used for computing the resonant frequency of A-shaped compact microstrip antennas (ACMAs) operating at UHF band. Firstly, the resonant frequencies of 144 ACMAs with various dimensions and electrical parameters were simulated with the help of IE3D™ based on method of moment (MoM). The ANN, ANFIS and SVM models for computing the resonant frequency were then built by considering the simulation data. 124 simulated ACMAs were utilized for training and the remaining 20 ACMAs were used for testing the ANN, ANFIS and SVM models. The performance of the ANN, ANFIS and SVM models are compared in the training and test process. The average percentage errors (APE) regarding the computed resonant frequencies for training of the ANN, ANFIS and SVM were obtained as 0.457%, 0.399% and 0.600%, respectively. The constructed models were then tested and APE values as 0.601% for ANN, 0.744% for ANFIS and 0.623% for SVM were achieved. The results obtained here show that ANN, ANFIS and SVM methods can be successfully applied to compute the resonant frequency of ACMAs, since they are useful and versatile methods that yield accurate results.

Keywords: a-shaped compact microstrip antenna, artificial neural network (ANN), adaptive neuro-fuzzy inference system (ANFIS), support vector machine (SVM)

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Authors: Dharmendra Pratap

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Large Cardamom (Amomum subulatum), family Zingiberaceae is an aromatic spice crop and has rich medicinal value. Large Cardamom is as synonymous to Sikkim as Tea is to Darjeeling. Since Sikkim alone contributes up to 88% of India's large cardamom production which is the world leader by producing over 50% of the global yield. However, the production of large cardamom has declined almost to half since last two decade. The economic losses have been attributed to two viral diseases namely, chirke and Foorkey. Chirke disease is characterized by light and dark green streaks on leaves. The affected leaves exhibit streak mosaic, which gradually coalesce, turn brown and eventually dry up. Excessive sprouting and formation of bushy dwarf clumps at the base of mother plants that gradually die characterize the foorkey disease. In our surveys in Sikkim–Darjeeling hill area during 2012-14, 40-45% of plants were found to be affected with foorkey disease and 10-15% with chirke. Mechanical and aphid transmission study showed banana as an alternate host for both the disease. For molecular identification, total genomic DNA and RNA was isolated from the infected leaf tissues and subjected to Rolling circle amplification (RCA) and RT-PCR respectively. The DNA concatamers produced in the RCA reaction were monomerized by different restriction enzymes and the bands corresponding to ~1 kb genomes were purified and cloned in the respective sites. The nucleotide sequencing results revealed the association of Nanovirus with the foorkey disease of large cardamom. DNA1 showed 74% identity with Replicase gene of FBNYV, DNA2 showed 77% identity with the NSP gene of BBTV and DNA3 showed 74% identity with CP gene of BBTV. The finding suggests the presence of a new species of nanovirus associated with foorkey disease of large cardamom in Sikkim and Darjeeling hills. The details of their epidemiology and other factors would be discussed.

Keywords: RCA, nanovirus, large cardamom, molecular virology and microbiology

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Authors: Jeanne Leblond, Warren Viricel, Amira Mbarek

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Although several products have reached the market, gene therapeutics are still in their first stages and require optimization. It is possible to improve their lacking efficiency by the use of carefully engineered vectors, able to carry the genetic material through each of the biological barriers they need to cross. In particular, getting inside the cell is a major challenge, because these hydrophilic nucleic acids have to cross the lipid-rich plasmatic and/or endosomal membrane, before being degraded into lysosomes. It takes less than one hour for newly endocytosed liposomes to reach highly acidic lysosomes, meaning that the degradation of the carried gene occurs rapidly, thus limiting the transfection efficiency. We propose to use a new pH-sensitive lipid able to change its conformation upon protonation at endosomal pH values, leading to the disruption of the lipidic bilayer and thus to the fast release of the nucleic acids into the cytosol. It is expected that this new pH-sensitive mechanism promote endosomal escape of the gene, thereby its transfection efficiency. The main challenge of this work was to design a preparation presenting fast-responding lipidic bilayer destabilization properties at endosomal pH 5 while remaining stable at blood pH value and during storage. A series of pH-sensitive lipids able to perform a conformational switch upon acidification were designed and synthesized. Liposomes containing these switchable lipids, as well as co-lipids were prepared and characterized. The liposomes were stable at 4°C and pH 7.4 for several months. Incubation with siRNA led to the full entrapment of nucleic acids as soon as the positive/negative charge ratio was superior to 2. The best liposomal formulation demonstrated a silencing efficiency up to 10% on HeLa cells, very similar to a commercial agent, with a lowest toxicity than the commercial agent. Using flow cytometry and microscopy assays, we demonstrated that drop of pH was required for the transfection efficiency, since bafilomycin blocked the transfection efficiency. Additional evidence was brought by the synthesis of a negative control lipid, which was unable to switch its conformation, and consequently exhibited no transfection ability. Mechanistic studies revealed that the uptake was mediated through endocytosis, by clathrin and caveolae pathways, as reported for previous lipid nanoparticle systems. This potent system was used for the treatment of hypercholesterolemia. The switchable lipids were able to knockdown PCSK9 expression on human hepatocytes (Huh-7). Its efficiency is currently evaluated on in vivo mice model of PCSK9 KO mice. In summary, we designed and optimized a new cationic pH-sensitive lipid for gene delivery. Its transfection efficiency is similar to the best available commercial agent, without the usually associated toxicity. The promising results lead to its use for the treatment of hypercholesterolemia on a mice model. Anticancer applications and pulmonary chronic disease are also currently investigated.

Keywords: liposomes, siRNA, pH-sensitive, molecular switch

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