Search results for: sickle cell anemia
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3701

Search results for: sickle cell anemia

941 In vitro Antioxidant Activity of Caesalpinia sappan Extract

Authors: Monthon Tangjitmungman

Abstract:

Numerous diseases have been linked to oxidative stress, in which a disproportion of free radicals in the body leads to tissue or cell damage. Polyphenols are the most abundant antioxidants found in plants, and they are highly effective at scavenging oxidative free radicals. Due to the presence of phenolic compounds in Caesalpinia sappan has been discovered to have antioxidant activity. It has several health benefits, the most important of which is preventing cardiovascular and cancer diseases. This study aimed to determine the phenolic content and antioxidant activity of C. sappan extract using a variety of antioxidant assays. The extract of C. sappan was made using a mixture of solvents (ethyl alcohol: water in ratio 8:2). The total phenolic content of C. sappan extract was determined and expressed as gallic acid equivalents using the Folin-Cioucalteu method (GAE). The antioxidant activity of C. sappan extract was assessed using the 2, 2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay and the ABTS radical scavenging capacity assay. An association was found between antioxidant activity and total phenol content. The antioxidant activity of C. sappan extract was also determined by DPPH and ABTS assays. The IC50 values for C. sappan extract from DPPH and ABTS assays were 54.48 μg/mL ± 0.545 and 25.46 μg/mL ± 0.790, respectively, in the DPPH assay. In the DPPH assay, vitamin C was used as a positive control, whereas Trolox was used as a positive control in the ABTS assay. In conclusion, C. sappan extract contains a high level of total phenolics and exhibits significant antioxidant activity. Nevertheless, more research should be done on the antioxidant activity, such as SOD and ROS scavenging assays and in vivo experiments, to determine whether the compound has antioxidant activity.

Keywords: ABTS assay, antioxidant activity, Caesalpinia sappan, DPPH assays, total phenolic content

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940 Screening the Growth Inhibition Mechanism of Sulfate-Reducing Bacteria by Chitosan/Lignosulfonate Nanocomposite in Seawater Media

Authors: K. Rasool

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Sulfate-reducing bacteria (SRBs) induced biofilm formation is a global industrial concern due to its role in the development of microbial-induced corrosion (MIC). Herein, we have developed a biodegradable chitosan/lignosulfonate nanocomposite (CS@LS) as an efficient green biocide for the inhibition of SRBs biofilms. We investigated in detail the inhibition mechanism of SRBs by CS@LS in seawater media. Stable CS@LS-1:1 with 150–200 nm average size and zeta potential of + 34.25 mV was synthesized. The biocidal performance of CS@LS was evaluated by sulfate reduction profiles coupled with analysis of extracted extracellular polymeric substances (EPS) and lactate dehydrogenase (LDH) release assays. As the nanocomposite concentration was increased from 50 to 500 µg/mL, the specific sulfate reduction rate (SSRR) decreased from 0.278 to 0.036 g-sulfate/g-VSS*day showing a relative sulfate reduction inhibition of 86.64% as compared to that of control. Similarly, the specific organic uptake rate (SOUR) decreased from 0.082 to 0.039 0.036 g-TOC/g-VSS*day giving a relative co-substrate oxidation inhibition of 52.19% as compared to that of control. The SRBs spiked with 500 µg/mL CS@LS showed a reduction in cell viability to 1.5 × 106 MPN/mL. To assess the biosafety of the nanocomposite on the marine biota, the 72-hours acute toxicity assays using the zebrafish embryo model revealed that the LC50 for the CS@LS was 103.3 µg/mL. Thus, CS@LS can be classified as environmentally friendly. The nanocomposite showed long-term stability and excellent antibacterial properties against SRBs growth and is thus potentially useful for combating the problems of biofilm growth in harsh marine and aquatic environments.

Keywords: green biocides, chitosan/lignosulfonate nanocomposite, SRBs, toxicity

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939 Multi-Objective Optimization of Intersections

Authors: Xiang Li, Jian-Qiao Sun

Abstract:

As the crucial component of city traffic network, intersections have significant impacts on urban traffic performance. Despite of the rapid development in transportation systems, increasing traffic volumes result in severe congestions especially at intersections in urban areas. Effective regulation of vehicle flows at intersections has always been an important issue in the traffic control system. This study presents a multi-objective optimization method at intersections with cellular automata to achieve better traffic performance. Vehicle conflicts and pedestrian interference are considered. Three categories of the traffic performance are studied including transportation efficiency, energy consumption and road safety. The left-turn signal type, signal timing and lane assignment are optimized for different traffic flows. The multi-objective optimization problem is solved with the cell mapping method. The optimization results show the conflicting nature of different traffic performance. The influence of different traffic variables on the intersection performance is investigated. It is observed that the proposed optimization method is effective in regulating the traffic at the intersection to meet multiple objectives. Transportation efficiency can be usually improved by the permissive left-turn signal, which sacrifices safety. Right-turn traffic suffers significantly when the right-turn lanes are shared with the through vehicles. The effect of vehicle flow on the intersection performance is significant. The display pattern of the optimization results can be changed remarkably by the traffic volume variation. Pedestrians have strong interference with the traffic system.

Keywords: cellular automata, intersection, multi-objective optimization, traffic system

Procedia PDF Downloads 557
938 Liquid Biopsy and Screening Biomarkers in Glioma Grading

Authors: Abdullah Abdu Qaseem Shamsan

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Background: Gliomas represent the most frequent, heterogeneous group of tumors arising from glial cells, characterized by difficult monitoring, poor prognosis, and fatality. Tissue biopsy is an established procedure for tumor cell sampling that aids diagnosis, tumor grading, and prediction of prognosis. We studied and compared the levels of liquid biopsy markers in patients with different grades of glioma. Also, it tried to establish the potential association between glioma and specific blood groups antigen. Result: 78 patients were identified, among whom maximum percentage with glioblastoma possessed blood group O+ (53.8%). The second highest frequency had blood group A+ (20.4%), followed by B+ (9.0%) and A- (5.1%), and least with O-. Liquid biopsy biomarkers comprised of ALT, LDH, lymphocytes, Urea, Alkaline phosphatase, AST Neutrophils, and CRP. The levels of all the components increased significantly with the severity of glioma, with maximum levels seen in glioblastoma (grade IV), followed by grade III and grade II respectively. Conclusion: Gliomas possess significant clinical challenges due to their progression with heterogeneous nature and aggressive behavior. Liquid biopsy is a non-invasive approach which aids to establish the status of the patient and determine the tumor grade, therefore may show diagnostic and prognostic utility. Additionally, our study provides evidence to demonstrate the role of ABO blood group antigens in the development of glioma. However, future clinical research on liquid biopsy will improve the sensitivity and specificity of these tests and validate their clinical usefulness to guide treatment approaches.

Keywords: GBM: glioblastoma multiforme, CT: computed tomography, MRI: magnetic resonance imaging, ctRNA: circulating tumor RNA

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937 Culturable Diversity of Halophilic Bacteria in Chott Tinsilt, Algeria

Authors: Nesrine Lenchi, Salima Kebbouche-Gana, Laddada Belaid, Mohamed Lamine Khelfaoui, Mohamed Lamine Gana

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Saline lakes are extreme hypersaline environments that are considered five to ten times saltier than seawater (150 – 300 g L-1 salt concentration). Hypersaline regions differ from each other in terms of salt concentration, chemical composition and geographical location, which determine the nature of inhabitant microorganisms. In order to explore the diversity of moderate and extreme halophiles Bacteria in Chott Tinsilt (East of Algeria), an isolation program was performed. In the first time, water samples were collected from the saltern during pre-salt harvesting phase. Salinity, pH and temperature of the sampling site were determined in situ. Chemical analysis of water sample indicated that Na +and Cl- were the most abundant ions. Isolates were obtained by plating out the samples in complex and synthetic media. In this study, seven halophiles cultures of Bacteria were isolated. Isolates were studied for Gram’s reaction, cell morphology and pigmentation. Enzymatic assays (oxidase, catalase, nitrate reductase and urease), and optimization of growth conditions were done. The results indicated that the salinity optima varied from 50 to 250 g L-1, whereas the optimum of temperature range from 25°C to 35°C. Molecular identification of the isolates was performed by sequencing the 16S rRNA gene. The results showed that these cultured isolates included members belonging to the Halomonas, Staphylococcus, Salinivibrio, Idiomarina, Halobacillus Thalassobacillus and Planococcus genera and what may represent a new bacterial genus.

Keywords: bacteria, Chott, halophilic, 16S rRNA

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936 Pale, Firm and Non-Exudative (PFN): An Emerging Major Broiler Breast Meat Group

Authors: Cintia Midori Kaminishikawahara, Fernanda Jéssica Mendonça, Moisés Grespan, Elza Iouko Ida, Massami Shimokomaki, Adriana Lourenço Soares

Abstract:

The quality of broiler breast meat is changing as a result of continuing emphasis on genetically bird’s selection for efficiently higher meat production. The consumer is experiencing a cooked product that is drier and less juicy when consumed. Breast meat has been classified as PSE (pale, soft, exudative), DFD (dark, firm, dry) and normal color meat. However, recently variations of this color have been observed and they are not in line with the specificity of the meat functional properties. Thus, the objective of this work was to report the finding of a new pale meat color group characterized as Pale, Firm and Non-exudative (PFN) based on its pH, color, meat functional properties and micro structural evaluation. Breast meat fillets samples (n=1045) from commercial line were classified into PSE (pH ≤5.8, L* ≥ 53.0), PFN (pH > 5.8 and L* ≥ 53.0) and Normal (pH >5.8 and L* < 53.0), based on pH and L* values. In sequence, a total of 30 samples of each group were analyzed for the water holding capacity (WHC) and shear force (SF). The incidence was 9.1% for PSE meat, 85.7% for PFN and 5.2% for Normal meat. The PSE meat presented lower values of WHC (P ≤ 0.05) followed in sequence by PFN and Normal samples and also the SF values of fresh PFN was higher than PSE meat (P ≤ 0.05) and similar to Normal samples. Under optical microscopy, the cell diameter was 10% higher for PFN in relation to PSE meat and similar to Normal meat. These preliminary results indicate an emerging group of breast meat and it should be considered that the Pale, Firm and Non-exudative should be considered as an ideal broiler breast meat quality.

Keywords: broiler PSE meat, light microscopy, texture, water holding capacity

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935 Anethum graveolens Prevents Liver and Kidney Injury, Oxidative Stress and Inflammation in Mice Exposed to Nicotine Perinatally

Authors: Saleh N. Maodaa

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Perinatal exposure to nicotine imbalances the redox status in newborns. This study investigated the effect of Anethum graveolens (dill) extract on oxidative stress and tissue injury in the liver and kidney of mice newborns exposed to nicotine perinatally. Pregnant mice received nicotine (0.25 mg/kg) on gestational day 12 to day 5 after birth and/or A. graveolens extract on a gestational day 1 to day 15 after birth. Newborn mice exposed to nicotine showed multiple histopathological alterations in the kidney and liver, including inflammatory cell infiltration and degenerative changes. Nicotine exposure increased hepatic and renal reactive oxygen species (ROS), lipid peroxidation, tumor necrosis factor (TNF-_), interleukin-6 (IL-6), and inducible nitric oxide synthase (iNOS) (p < 0.001), and decreased antioxidant defenses (p < 0.001). A. graveolens supplementation significantly prevented liver and kidney injury, suppressed ROS generation (p < 0.001), lipid peroxidation (p < 0.001), and inflammatory response (p < 0.001), and enhanced antioxidant defenses. In addition, A. graveolens upregulated hepatic and renal Nrf2 and HO-1 mRNA and increased HO-1 activity in normal and nicotine-exposed mice. In conclusion, A. graveolens protects against perinatal nicotine-induced oxidative stress, inflammation, and tissue injury in the liver and kidney of newborn mice. A. graveolens upregulated hepatic and renal Nrf2/HO-1 signaling and enhanced antioxidant defenses in mice.

Keywords: dill, oxidative stress, cytokines, nicotine

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934 Study of Age-Dependent Changes of Peripheral Blood Leukocytes Apoptotic Properties

Authors: Anahit Hakobjanyan, Zdenka Navratilova, Gabriela Strakova, Martin Petrek

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Aging has a suppressive influence on human immune cells. Apoptosis may play important role in age-dependent immunosuppression and lymphopenia. Prevention of apoptosis may be promoted by BCL2-dependent and BCL2-independent manner. BCL2 is an antiapoptotic factor that has an antioxidative role by locating the glutathione at mitochondria and repressing oxidative stress. STAT3 may suppress apoptosis in BCL2-independent manner and promote cell survival blocking cytochrome-c release and reducing ROS production. The aim of our study was to estimate the influence of aging on BCL2-dependent and BCL2-independent prevention of apoptosis via measurement of BCL2 and STAT3 mRNAs expressions. The study was done on Armenian population (2 groups: 37 healthy young (mean age±SE; min/max age, male/female: 37.6±1.1; 20/54, 15/22), 28 healthy aged (66.7±1.5; 57/85, 12/16)). mRNA expression in peripheral blood leukocytes (PBL) was determined by RT-PCR using PSMB2 as the reference gene. Statistical analysis was done with Graph-Pad Prism 5; P < 0.05 considered as significant. The expression of BCL2 mRNA was lower in aged group (0.199) compared with young ones (0.643)(p < 0.01). Decrease expression was also recorded for female and male subgroups (p < 0.01). The expression level of STAT3 mRNA was increased (young, 0.228; aged, 0.428) (p < 0.05) during aging (in the whole age group and male/female subgroups). Decreased level of BCL2 mRNA may indicate about the suppression of BCL2-dependent prevention of apoptosis during aging in peripheral blood leukocytes. At the same time increased the level of STAT3 may suggest about activation of BCL2-independent prevention of apoptosis during aging.

Keywords: BCL2, STAT3, aging, apoptosis

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933 Observation of the Orthodontic Tooth's Long-Term Movement Using Stereovision System

Authors: Hao-Yuan Tseng, Chuan-Yang Chang, Ying-Hui Chen, Sheng-Che Chen, Chih-Han Chang

Abstract:

Orthodontic tooth treatment has demonstrated a high success rate in clinical studies. It has been agreed upon that orthodontic tooth movement is based on the ability of surrounding bone and periodontal ligament (PDL) to react to a mechanical stimulus with remodeling processes. However, the mechanism of the tooth movement is still unclear. Recent studies focus on the simple principle compression-tension theory while rare studies directly measure tooth movement. Therefore, tracking tooth movement information during orthodontic treatment is very important in clinical practice. The aim of this study is to investigate the mechanism responses of the tooth movement during the orthodontic treatments. A stereovision system applied to track the tooth movement of the patient with the stamp brackets. The system was established by two cameras with their relative position calibrate. And the orthodontic force measured by 3D printing model with the six-axis load cell to determine the initial force application. The result shows that the stereovision system accuracy revealed the measurement presents a maximum error less than 2%. For the study on patient tracking, the incisor moved about 0.9 mm during 60 days tracking, and half of movement occurred in the first few hours. After removing the orthodontic force in 100 hours, the distance between before and after position incisor tooth decrease 0.5 mm consisted with the release of the phenomenon. Using the stereovision system can accurately locate the three-dimensional position of the teeth and superposition of 3D coordinate system for all the data to integrate the complex tooth movement.

Keywords: orthodontic treatment, tooth movement, stereovision system, long-term tracking

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932 Understanding the Mechanisms of Salmonella typhimurium Resistance to Cannabidiol

Authors: Iddrisu Ibrahim, Joseph Atia Ayariga, Junhuan Xu, Daniel Abugri, Boakai Robertson, Olufemi S. Ajayi

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The emergence of multidrug resistance poses a huge risk to public health globally. Yet these recalcitrant pathogens continue to rise in incidence rate, with resistance rates significantly outpacing the speed of antibiotic development. This, therefore, presents an aura of related health issues such as untreatable nosocomial infections arising from organ transplants and surgeries, as well as community-acquired infections that are related to people with compromised immunity, e.g., diabetic and HIV patients, etc. There is a global effort to fight multidrug-resistant pathogens spearheaded by the World Health Organization, thus calling for research into novel antimicrobial agents to fight multiple drug resistance. Previously, our laboratory demonstrated that Cannabidiol (CBD) was an effective antimicrobial against Salmonella typhimurium (S. typhimurium). However, we observed resistance development over time. To understand the mechanisms S. typhimurium uses to develop resistance to Cannabidiol (CBD), we studied the abundance of bacteria lipopolysaccharide (LPS) and membrane sterols of both susceptible and resistant S. typhimurium. Using real-time quantitative polymerase chain reaction (RT-qPCR), we also analyzed the expression of selected genes known for aiding resistance development in S. typhimurium. We discovered that there was a significantly higher expression of blaTEM, fimA, fimZ, and integrons in the CBD-resistant bacteria, and these were also accompanied by a shift in abundance in cell surface molecules such as lipopolysaccharide (LPS) and sterols.

Keywords: antimicrobials, resistance, cannabidiol, gram-negative bacteria, integrons, blaTEM, Fim, LPS, ergosterols

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931 Chemical Composition and Biological Investigation of Halpophyllum tuberculatum A. Juss (Rutaceae) Essential Oils Growing in Libya

Authors: O. M. M. Sabry, Abeer M. El Sayed

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The essential oils from the aerial parts and flowers of Haplophyllum tuberculatum (Forsskal) Adr. Juss (Rutaceae) growing in Libya were obtained separately by hydro-distillation using a Clevenger-type apparatus. The essential oils yield were (0.4, 1.5w/w%) respectively based on the dry weight of the plant. The oils were analyzed by GC-MS. Twenty four constituents, amounting to 96.6%, were identified in the oil of the aerial parts. The predominant compounds were among the non oxygenated terpenoids (82.4%) as monoterpene hydrocarbons, represented by sabinen (26.4 %), δ-terpinen (26 %), β-phellandrene (10.4%) and 3-carene (3.86%). Zingiberine (0.4%) and β-sesquiphellandrene (0.12%) were the major sesquiterpene hydrocarbons identified. Oxygenated monoterpenes were represented by eucalyptol (5.5%) and piperitone (5.55%). Twenty six constituents, equivalent to 99.5%, were identified in the oil of the flowers. The dominance of monoterpene hydrocarbons in the flowers oil can be attributed to the high percentage of γ-terpinen (38.44%), β-phellandrene (10.0%), α- phellandrene (2.33%), 3,4-dimethyl-1,5-cyclooctadiene (6.67%), β-myrecene (6.04%), 3-carene (5.43%) and α-pinene (1.3%).While the oxygenated monoterpenes can be contributed to the trans-piperitol (4.67%) and piperitone (2.07%). Sesquiterpene hydrocarbons were not identified in the oil of the flower of H. tuberculatum. Variation in constitution between oils of Libyan H. tuberculatum and that obtained from other countries can be due to both environmental and genetic factors. The essential oils have demonstrated variable antimicrobial activities against certain micro-organisms. Also have revealed marked in vitro cytotoxicity against lung (H1299), liver (HEPG2) carcinoma cell line and variably effective as anti-inflammatory and antioxidant.

Keywords: Halpophyllum tuberculatum, rutaceae, essential oil, antimicrobial, anti-inflammatory, antitumor, antioxidant, Libya

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930 Role of Biomaterial Surface Nanotopography on Protein Unfolding and Immune Response

Authors: Rahul Madathiparambil Visalakshan, Alex Cavallaro, John Hayball, Krasimir Vasilev

Abstract:

The role of biomaterial surface nanotopograhy on fibrinogen adsorption and unfolding, and the subsequent immune response were studied. Inconsistent topography and varying chemical functionalities along with a lack of reproducibility pose a challenge in determining the specific effects of nanotopography or chemistry on proteins and cells. It is important to have a well-defined nanotopography with a homogeneous chemistry to study the real effect of nanotopography on biological systems. Therefore, we developed a technique that can produce well-defined and highly reproducible topography to identify the role of specific roughness, size, height and density with the presence of homogeneous chemical functionality. Using plasma polymerisation of oxazoline monomers and immobilized gold nanoparticles we created surfaces with an equal number density of nanoparticles of different sizes. This surface was used to study the role of surface nanotopography and the interplay of surface chemistry on proteins and immune cells. The effect of nanotopography on fibrinogen adsorption was investigated using Quartz Cristal Microbalance with Dissipation and micro BCA. The mass of fibrinogen adsorbed on the surface increased with increasing size of nano-topography. Protein structural changes up on adsorption to the nano rough surface was studied using circular dichroism spectroscopy. Fibrinogen unfolding varied depending on the specific nanotopography of the surfaces. It was revealed that the in vitro immune response to the nanotopography surfaces changed due to this protein unfolding.

Keywords: biomaterial inflammation, protein and cell responses, protein unfolding, surface nanotopography

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929 Genetics of Atopic Dermatitis: Role of Cytokine Genes Polymorphisms

Authors: Ghaleb Bin Huraib

Abstract:

Atopic dermatitis (AD), also known as atopic eczema, is a chronic inflammatory skin disease characterized by severe itching and recurrent, relapsing eczema-like skin lesions, affecting up to 20% of children and 10% of adults in industrialized countries. AD is a complex multifactorial disease, and its exact etiology and pathogenesis have not been fully elucidated. The aim of this study was to investigate the impact of gene polymorphisms of T helper cell subtype Th1 and Th2 cytokines, interferon-gamma (IFN-γ), interleukin-6 (IL-6) and transforming growth factor (TGF)-β1on AD susceptibility in a Saudi cohort. One hundred four unrelated patients with AD and 195 healthy controls were genotyped for IFN-γ (874A/T), IL-6 (174G/C) and TGF-β1 (509C/T) polymorphisms using ARMS-PCR and PCR-RFLP technique. The frequency of genotypes AA and AT of IFN-γ (874A/T) differed significantly among patients and controls (P 0.001). The genotype AT was increased while genotype AA was decreased in AD patients as compared to controls. AD patients also had a higher frequency of T-containing genotypes (AT+TT) than controls (P = 0.001). The frequencies of alleles T and A were statistically different in patients and controls (P = 0.04). The frequencies of genotype GG and allele G of IL-6 (174G/C) were significantly higher, while genotype GC and allele C were lower in AD patients than in controls. There was no significant difference in the frequencies of alleles and genotypes of TGF-β1 (509C/T) polymorphism between the patient and control groups. These results showed that susceptibility to AD is influenced by the presence or absence of genotypes of IFN-γ (874A/T) and IL-6 (174G/C) polymorphisms. It is concluded T-allele and T-containing genotypes (AT+TT) of IFN-γ (874A/T) and G-allele and GG genotype ofIL-6 (174G/C) polymorphisms are susceptible to AD in Saudis. On the other hand, the TGF-β1 (509C/T) polymorphism may not be associated with AD risk in our population; however, further studies with large sample sizes are required to confirm these results.

Keywords: atopic dermatitis, Polymorphism, Interferon, IL-6

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928 Impact of Edible Coatings Made of Chitosan and Spray Dried Propolis in the Shell Life of White Cachama (Piaractus brachypomus)

Authors: David Guillermo Piedrahita Marquez, Hector Suarez Mahecha, Jairo Humberto Lopez

Abstract:

There is a need to preserve aquaculture matrices due to their high nutritional value, and its broad consumption, one of those species is the white cachama (Piaractus brachypomus), this fish is located in the rivers of eastern Colombia, and the previously mentioned species needs more study. Therefore, in a paper the effects of an alternative method of preservation of shell life were investigated, the method used is the application of an edible coating made from chitosan and ethanolic extract of propolis (EEP) encapsulated in maltodextrin. The coating was applied by immersion, and after that, we investigated the post mortem quality changes of the fish performing physicochemical and microbiological analysis. pH, volatile bases, test thiobarbituric acid and peroxide value were tested; finally, we studied the effect of the coating on mesophilic strains, coliforms and other microorganisms such as Staphylococcus, and Salmonella. Finally, we concluded that the coating prolongs the shelf life because it acts as a barrier to oxygen and moisture, the bioactive compounds trap free radicals and the coatings changes the metabolism and cause the cell lysis of the microorganisms. It was determined that the concentration of malonaldehyde, the volatile basic nitrogen content and pH are the variables that distinguish more clearly between the samples with the treatment and the control samples.

Keywords: antimicrobial activity, lipid oxidation, texture profile analysis (TPA), sensorial analysis, peroxide value, thiobarbituric acid assay (TBA), total volatile basic nitrogen (TVB-N)

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927 Unsupervised Segmentation Technique for Acute Leukemia Cells Using Clustering Algorithms

Authors: N. H. Harun, A. S. Abdul Nasir, M. Y. Mashor, R. Hassan

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Leukaemia is a blood cancer disease that contributes to the increment of mortality rate in Malaysia each year. There are two main categories for leukaemia, which are acute and chronic leukaemia. The production and development of acute leukaemia cells occurs rapidly and uncontrollable. Therefore, if the identification of acute leukaemia cells could be done fast and effectively, proper treatment and medicine could be delivered. Due to the requirement of prompt and accurate diagnosis of leukaemia, the current study has proposed unsupervised pixel segmentation based on clustering algorithm in order to obtain a fully segmented abnormal white blood cell (blast) in acute leukaemia image. In order to obtain the segmented blast, the current study proposed three clustering algorithms which are k-means, fuzzy c-means and moving k-means algorithms have been applied on the saturation component image. Then, median filter and seeded region growing area extraction algorithms have been applied, to smooth the region of segmented blast and to remove the large unwanted regions from the image, respectively. Comparisons among the three clustering algorithms are made in order to measure the performance of each clustering algorithm on segmenting the blast area. Based on the good sensitivity value that has been obtained, the results indicate that moving k-means clustering algorithm has successfully produced the fully segmented blast region in acute leukaemia image. Hence, indicating that the resultant images could be helpful to haematologists for further analysis of acute leukaemia.

Keywords: acute leukaemia images, clustering algorithms, image segmentation, moving k-means

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926 Electrochemical Behavior of Cocaine on Carbon Paste Electrode Chemically Modified with Cu(II) Trans 3-MeO Salcn Complex

Authors: Alex Soares Castro, Matheus Manoel Teles de Menezes, Larissa Silva de Azevedo, Ana Carolina Caleffi Patelli, Osmair Vital de Oliveira, Aline Thais Bruni, Marcelo Firmino de Oliveira

Abstract:

Considering the problem of the seizure of illicit drugs, as well as the development of electrochemical sensors using chemically modified electrodes, this work shows the study of the electrochemical activity of cocaine in carbon paste electrode chemically modified with Cu (II) trans 3-MeO salcn complex. In this context, cyclic voltammetry was performed on 0.1 mol.L⁻¹ KCl supporting electrolyte at a scan speed of 100 mV s⁻¹, using an electrochemical cell composed of three electrodes: Ag /AgCl electrode (filled KCl 3 mol.L⁻¹) from Metrohm® (reference electrode); a platinum spiral electrode, as an auxiliary electrode, and a carbon paste electrode chemically modified with Cu (II) trans 3-MeO complex (as working electrode). Two forms of cocaine were analyzed: cocaine hydrochloride (pH 3) and cocaine free base form (pH 8). The PM7 computational method predicted that the hydrochloride form is more stable than the free base form of cocaine, so with cyclic voltammetry, we found electrochemical signal only for cocaine in the form of hydrochloride, with an anodic peak at 1.10 V, with a linearity range between 2 and 20 μmol L⁻¹ had LD and LQ of 2.39 and 7.26x10-5 mol L⁻¹, respectively. The study also proved that cocaine is adsorbed on the surface of the working electrode, where through an irreversible process, where only anode peaks are observed, we have the oxidation of cocaine, which occurs in the hydrophilic region due to the loss of two electrons. The mechanism of this reaction was confirmed by the ab-inito quantum method.

Keywords: ab-initio computational method, analytical method, cocaine, Schiff base complex, voltammetry

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925 Stem Cell Augmentation Therapy for Cardiovascular Risk in Ankylosing Spondylitis: STATIN-as Study

Authors: Ashit Syngle, Nidhi Garg, Pawan Krishan

Abstract:

Objective: Bone marrow derived stem cells, endothelial progenitor cells (EPCs), protect against atherosclerotic vascular damage. However, EPCs are depleted in AS and contribute to the enhanced cardiovascular risk. Statins have a protective effect in CAD and diabetes by enhancing the proliferation, migration and survival of EPCs. Therapeutic potential of augmenting EPCs to treat the heightened cardiovascular risk of AS has not yet been exploited. We aimed to investigate the effect of rosuvastatin on EPCs population and inflammation in AS. Methods: 30 AS patients were randomized to receive 6 months of treatment with rosuvastatin (10 mg/day, n=15) and placebo (n=15) as an adjunct to existing stable anti-rheumatic drugs. EPCs (CD34+/CD133+) were quantified by Flow Cytometry. Inflammatory measures (BASDAI, BASFI, CRP and ESR), pro-inflammatory cytokines (TNF-α, IL-6 and IL-1) and lipids were measured at baseline and after treatment. Results: At baseline, inflammatory measures and pro-inflammatory cytokines were elevated and EPCs depleted among both groups. EPCs increased significantly (p < 0.01) after treatment with rosuvastatin. At 6 months, BASDAI, BASFI, ESR, CRP, TNF-α, and IL-6 improved significantly in rosuvastatin group. Significant negative correlation was observed between EPCs and BASDAI, CRP and IL-6 after rosuvastatin treatment. Conclusion: First study to show that rosuvastatin augments EPCs population in AS. This defines a novel mechanism of rosuvastatin treatment in AS: the augmentation of EPCs with improvement in proinflammatory cytokines and inflammatory disease activity. The augmentation of EPCs by rosuvastatin may provide a novel strategy to prevent cardiovascular events in AS.

Keywords: ankylosing spondylitis, Endothelial Progenitor Cells, inflammation, pro-inflammatory cytokines, rosuvastatin

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924 Effect of Barium Doping on Structural, Morphological, Optical, and Photocatalytic Properties of Sprayed ZnO Thin Films

Authors: Halima Djaaboube, Redha Aouati, Ibtissem Loucif, Yassine Bouachiba, Mouad Chettab, Adel Taabouche, Sihem Abed, Salima Ouendadji, Abderrahmane Bouabellou

Abstract:

Thin films of pure and barium-doped zinc oxide (ZnO) were prepared using spray pyrolysis process. The films were deposited on glass substrates at 450°C. The different samples are characterized by X-ray diffraction (XRD) and UV-Vis spectroscopy. X-ray diffraction patterns reveal the formation of a single ZnO Wurtzite structure and the good crystallinity of the films. The substitution of Ba ions influences the texture of the layers and makes the (002) plane a preferential growth plane. At concentrations below 6% Ba, the hexagonal structure of ZnO undergoes compressive stresses due to barium ions which have a radius twice of the Zn ions. This result leads to the decrees of a and c parameters and therefore the volume of the unit cell. This result is confirmed by the decrease in the number of crystallites and the increase in the size of the crystallites. At concentrations above 6%, barium substitutes the zinc atom and modifies the structural parameters of the thin layers. The bandgap of ZnO films decreased with increasing doping, this decrease is probably due to the 4d orbitals of the Ba atom due to the sp-d spin-exchange interactions between the band electrons and the localized d-electrons of the substituted Ba ion. Although, the Urbache energy undergoes an increase which implies the creation of energy levels below the conduction band and decreases the band gap width. The photocatalytic activity of ZnO doped 9% Ba was evaluated by the photodegradation of methylene blue under UV irradiation.

Keywords: barium, doping, photodegradation, spray pyrolysis, ZnO.

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923 Algebraic Coupled Level Set-Volume of Fluid Method with Capillary Pressure Treatment for Surface Tension Dominant Two-Phase Flows

Authors: Majid Haghshenas, James Wilson, Ranganathan Kumar

Abstract:

In this study, an Algebraic Coupled Level Set-Volume of Fluid (A-CLSVOF) method with capillary pressure treatment is proposed for the modeling of two-phase capillary flows. The Volume of Fluid (VOF) method is utilized to incorporate one-way coupling with the Level Set (LS) function in order to further improve the accuracy of the interface curvature calculation and resulting surface tension force. The capillary pressure is determined and treated independently of the hydrodynamic pressure in the momentum balance in order to maintain consistency between cell centered and interpolated values, resulting in a reduction in parasitic currents. In this method, both VOF and LS functions are transported where the new volume fraction determines the interface seed position used to reinitialize the LS field. The Hamilton-Godunov function is used with a second order (in space and time) discretization scheme to produce a signed distance function. The performance of the current methodology has been tested against some common test cases in order to assess the reduction in non-physical velocities and improvements in the interfacial pressure jump. The cases of a static drop, non-linear Rayleigh-Taylor instability and finally a droplets impact on a liquid pool were simulated to compare the performance of the present method to other well-known methods in the area of parasitic current reduction, interface location evolution and overall agreement with experimental results.

Keywords: two-phase flow, capillary flow, surface tension force, coupled LS with VOF

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922 Quorum-Sensing Driven Inhibitors for Mitigating Microbial Influenced Corrosion

Authors: Asma Lamin, Anna H. Kaksonen, Ivan Cole, Paul White, Xiao-Bo Chen

Abstract:

Microbiologically influenced corrosion (MIC) is a process in which microorganisms initiate, facilitate, or accelerate the electrochemical corrosion reactions of metallic components. Several reports documented that MIC accounts for about 20 to 40 % of the total cost of corrosion. Biofilm formation due to the presence of microorganisms on the surface of metal components is known to play a vital role in MIC, which can lead to severe consequences in various environmental and industrial settings. Quorum sensing (QS) system plays a major role in regulating biofilm formation and control the expression of some microbial enzymes. QS is a communication mechanism between microorganisms that involves the regulation of gene expression as a response to the microbial cell density within an environment. This process is employed by both Gram-positive and Gram-negative bacteria to regulate different physiological functions. QS involves production, detection, and responses to signalling chemicals, known as auto-inducers. QS controls specific processes important for the microbial community, such as biofilm formation, virulence factor expression, production of secondary metabolites and stress adaptation mechanisms. The use of QS inhibitors (QSIs) has been proposed as a possible solution to biofilm related challenges in many different applications. Although QSIs have demonstrated some strength in tackling biofouling, QSI-based strategies to control microbially influenced corrosion have not been thoroughly investigated. As such, our research aims to target the QS mechanisms as a strategy for mitigating MIC on metal surfaces in engineered systems.

Keywords: quorum sensing, quorum quenching, biofilm, biocorrosion

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921 Iron Catalyst for Decomposition of Methane: Influence of Al/Si Ratio Support

Authors: A. S. Al-Fatesh, A. A. Ibrahim, A. M. AlSharekh, F. S. Alqahtani, S. O. Kasim, A. H. Fakeeha

Abstract:

Hydrogen is the expected future fuel since it produces energy without any pollution. It can be used as a fuel directly or through the fuel cell. It is also used in chemical and petrochemical industry as reducing agent or in hydrogenation processes. It is produced by different methods such as reforming of hydrocarbon, electrolytic method and methane decomposition. The objective of the present paper is to study the decomposition of methane reaction at 700°C and 800°C. The catalysts were prepared via impregnation method using 20%Fe and different proportions of combined alumina and silica support using the following ratios [100%, 90%, 80%, and 0% Al₂O₃/SiO₂]. The prepared catalysts were calcined and activated at 600 OC and 500 OC respectively. The reaction was carried out in fixed bed reactor at atmospheric pressure using 0.3g of catalyst and feed gas ratio of 1.5/1 CH₄/N₂ with a total flow rate 25 mL/min. Catalyst characterizations (TPR, TGA, BET, XRD, etc.) have been employed to study the behavior of catalysts before and after the reaction. Moreover, a brief description of the weight loss and the CH₄ conversions versus time on stream relating the different support ratios over 20%Fe/Al₂O₃/SiO₂ catalysts has been added as well. The results of TGA analysis provided higher weights losses for catalysts operated at 700°C than 800°C. For the 90% Al₂O₃/SiO₂, the activity decreases with the time on stream using 800°C reaction temperature from 73.9% initial CH₄ conversion to 46.3% for a period of 300min, whereas the activity for the same catalyst increases from 47.1% to 64.8% when 700°C reaction temperature is employed. Likewise, for 80% Al₂O₃/SiO₂ the trend of activity is similar to that of 90% Al₂O₃/SiO₂ but with a different rate of activity variation. It can be inferred from the activity results that the ratio of Al₂O₃ to SiO₂ is crucial and it is directly proportional with the activity. Whenever the Al/Si ratio decreases the activity declines. Indeed, the CH₄ conversion of 100% SiO₂ support was less than 5%.

Keywords: Al₂O₃, SiO₂, CH₄ decomposition, hydrogen, iron

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920 InP Nanocrystals Core and Surface Electronic Structure from Ab Initio Calculations

Authors: Hamad R. Jappor, Zeyad Adnan Saleh, Mudar A. Abdulsattar

Abstract:

The ab initio restricted Hartree-Fock method is used to simulate the electronic structure of indium phosphide (InP) nanocrystals (NCs) (216-738 atoms) with sizes ranging up to about 2.5 nm in diameter. The calculations are divided into two parts, surface, and core. The oxygenated (001)-(1×1) facet that expands with larger sizes of nanocrystals is investigated to determine the rule of the surface in nanocrystals electronic structure. Results show that lattice constant and ionicity of the core part show decreasing order as nanocrystals grow up in size. The smallest investigated nanocrystal is 1.6% larger in lattice constant and 131.05% larger in ionicity than the converged value of largest investigated nanocrystal. Increasing nanocrystals size also resulted in an increase of core cohesive energy (absolute value), increase of core energy gap, and increase of core valence. The surface states are found mostly non-degenerated because of the effect of surface discontinuity and oxygen atoms. Valence bandwidth is wider on the surface due to splitting and oxygen atoms. The method also shows fluctuations in the converged energy gap, valence bandwidth and cohesive energy of core part of nanocrystals duo to shape variation. The present work suggests the addition of ionicity and lattice constant to the quantities that are affected by quantum confinement phenomenon. The method of the present model has threefold results; it can be used to approach the electronic structure of crystals bulk, surface, and nanocrystals.

Keywords: InP, nanocrystals core, ionicity, Hartree-Fock method, large unit cell

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919 Numerical Analysis of Core-Annular Blood Flow in Microvessels at Low Reynolds Numbers

Authors: L. Achab, F. Iachachene

Abstract:

In microvessels, red blood cells (RBCs) exhibit a tendency to migrate towards the vessel center, establishing a core-annular flow pattern. The core region, marked by a high concentration of RBCs, is governed by significantly non-Newtonian viscosity. Conversely, the annular layer, composed of cell-free plasma, is characterized by Newtonian low viscosity. This property enables the plasma layer to act as a lubricant for the vessel walls, efficiently reducing resistance to the movement of blood cells. In this study, we investigate the factors influencing blood flow in microvessels and the thickness of the annular plasma layer using a non-miscible fluids approach in a 2D axisymmetric geometry. The governing equations of an incompressible unsteady flow are solved numerically through the Volume of Fluid (VOF) method to track the interface between the two immiscible fluids. To model blood viscosity in the core region, we adopt the Quemada constitutive law which is accurately captures the shear-thinning blood rheology over a wide range of shear rates. Our results are then compared to an established theoretical approach under identical flow conditions, particularly concerning the radial velocity profile and the thickness of the annular plasma layer. The simulation findings for low Reynolds numbers, demonstrate a notable agreement with the theoretical solution, emphasizing the pivotal role of blood’s rheological properties in the core region in determining the thickness of the annular plasma layer.

Keywords: core-annular flows, microvessels, Quemada model, plasma layer thickness, volume of fluid method

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918 Hematological Changes in Oral Cancer Patients with Smokable and Chewable Tobacco

Authors: Mohsin Ali Baloch, Saira Baloch

Abstract:

Objective: To analyze hematological changes in patients of oral cancer with history of smokable and chewable tobacco use, and to compare them with healthy controls. Study Design: Descriptive type of study survey. Setting: This study was conducted at the Department of Oral and Maxillofacial Surgery, LUMHS, Jamshoro. Study Period: One year July, 2013 to July, 2014. Subject and Methods: Histopathologically confirmed hundred cases of oral cancer with the history of smokable and non-smokable tobacco were selected to analyze the hematological variation. Inclusion Criteria: Histopathologically diagnosed patients of oral squamous cell carcinoma, with history of smokable and non-smokable tobacco. Exclusion Criteria: Patient with any systemic medically compromising problem, terminally ill patients, radio or chemotherapeutically treated patients, patients with metastasis to lungs or any distant metastasis, patients with the history of more than one well-defined etiological factor involved. Results: There were 73% patients of oral cancer reported with anemic. Significantly lower values of Hb, platelet, and higher mean values of ESR, TLC, and were observed in both groups of oral cancer patients; tobacco smokers and tobacco chewers as compared to non-smokers healthy controls. There was more decline in the level of haemoglobin and incline in the level of ESR observed in tobacco chewer oral cancer patients as compared to tobacco smokers patients, while TLC was more observed in smokers. Conclusion: Oral cancer patients with a history of chewable/smokable tobacco have likely worse hematological profile, which increases the anesthetic and surgical challenges for maxillofacial surgeons, which have a significant impact on treatment planning as well.

Keywords: oral cancer, hematological variations, tobacco, smokers

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917 Membrane-Localized Mutations as Predictors of Checkpoint Blockade Efficacy in Cancer

Authors: Zoe Goldberger, Priscilla S. Briquez, Jeffrey A. Hubbell

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Tumor cells have mutations resulting from genetic instability that the immune system can actively recognize. Immune checkpoint immunotherapy (ICI) is commonly used in the clinic to re-activate immune reactions against mutated proteins, called neoantigens, resulting in tumor remission in cancer patients. However, only around 20% of patients show durable response to ICI. While tumor mutational burden (TMB) has been approved by the Food and Drug Administration (FDA) as a criterion for ICI therapy, the relevance of the subcellular localizations of the mutated proteins within the tumor cell has not been investigated. Here, we hypothesized that localization of mutations impacts the effect of immune responsiveness to ICI. We analyzed publicly available tumor mutation sequencing data of ICI treated patients from 3 independent datasets. We extracted the subcellular localization from the UniProtKB/Swiss-Prot database and quantified the proportion of membrane, cytoplasmic, nuclear, or secreted mutations per patient. We analyzed this information in relation to response to ICI treatment and overall survival of patients showing with 1722 ICI-treated patients that high mutational burden localized at the membrane (mTMB), correlate with ICI responsiveness, and improved overall survival in multiple cancer types. We anticipate that our results will ameliorate predictability of cancer patient response to ICI with potential implications in clinical guidelines to tailor ICI treatment. This would not only increase patient survival for those receiving ICI, but also patients’ quality of life by reducing the number of patients enduring non-effective ICI treatments.

Keywords: cancer, immunotherapy, membrane neoantigens, efficacy prediction, biomarkers

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916 Laser-Hole Boring into Overdense Targets: A Detailed Study on Laser and Target Properties

Authors: Florian Wagner, Christoph Schmidt, Vincent Bagnoud

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Understanding the interaction of ultra-intense laser pulses with overcritical targets is of major interest for many applications such as laser-driven ion acceleration, fast ignition in the frame of inertial confinement fusion or high harmonic generation and the creation of attosecond pulses. One particular aspect of this interaction is the shift of the critical surface, where the laser pulse is stopped and the absorption is at maximum, due to the radiation pressure induced by the laser pulse, also referred to as laser hole boring. We investigate laser-hole boring experimentally by measuring the backscattered spectrum which is doppler-broadened because of the movement of the reflecting surface. Using the high-power, high-energy laser system PHELIX in Darmstadt, we gathered an extensive set of data for different laser intensities ranging from 10^18 W/cm2 to 10^21 W/cm2, two different levels of the nanosecond temporal contrast (10^6 vs. 10^11), elliptical and linear polarization and varying target configurations. In this contribution we discuss how the maximum velocity of the critical surface depends on these parameters. In particular we show that by increasing the temporal contrast the maximum hole boring velocity is decreased by more than a factor of three. Our experimental findings are backed by a basic analytical model based on momentum and mass conservation as well as particle in cell simulations. These results are of particular importance for fast ignition since they contribute to a better understanding of the transport of the ignitor pulse into the overdense region.

Keywords: laser-hole boring, interaction of ultra-intense lasers with overcritical targets, fast ignition, relativistic laser motter interaction

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915 Circadian Expression of MicroRNAs in Colon and Its Changes during Colorectal Tumorigenesis

Authors: Katerina Balounova, Jiri Pacha, Peter Ergang, Martin Vodicka, Pavlina Kvapilova

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MicroRNAs are small non-coding RNAs involved in a wide range of physiological processes. Post-transcriptional regulation of gene expression by microRNAs gives the organism a further level of control of the gene-expression program and the disruption of this microRNA regulatory mechanism seems to increase the risk of various pathophysiological conditions including tumorigenesis. To the present day, microRNAs were shown to participate in the mayor signalization pathways leading to tumorigenesis, including proliferation, cell cycle, apoptosis and metastasis formation. In addition, microRNAs have been found to play important roles in the generation and maintenance of circadian clock. These clocks generate circadian rhythms, which participate in a number of regulatory pathways. Disruption of the circadian signals seems to be associated with the development and the progression of tumours including colorectal cancer. We investigated therefore whether the diurnal profiles of miRNAs linked to tumorigenesis and regulation of circadian clock are changed during tumorigenesis. Based on published data we chose 10 microRNAs linked to tumorigenesis or circadian clock (let-7b-5p, miR 1 3p, miR 106b 5p, miR 141 3p, miR 191 5p, miR 20a 5p, miR 25 3p, miR 29a 3p, miR 34a 5p and miR 93 5p) and compared their 24-hr expression profiles in healthy and in chemically induces primary colorectal tumours of 52week-old mice. Using RT-qPCR we proved circadian rhythmicity in let-7b-5p, miR 106b 5p, miR 141 3p, miR 191 5p, miR 20a 5p, miR 25 3p, miR 29a 3p and miR 93 5p in healthy colon but not in tumours. The acrophases of miR 106b 5p, miR 141 3p, miR 191 5p, miR 20a 5p, miR 25 3p and miR 93 5p were reached around CT 24, the acrophases of let-7b-5p and miR-29a-3p were slightly shifted and reached around CT 21. In summary, our results show that circadian regulation of some colonic microRNAs is greatly affected by neoplastic transformation.

Keywords: circadian rhythm, colon, colorectal cancer, microRNA, tumorigenesis

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914 Bluetooth Communication Protocol Study for Multi-Sensor Applications

Authors: Joao Garretto, R. J. Yarwood, Vamsi Borra, Frank Li

Abstract:

Bluetooth Low Energy (BLE) has emerged as one of the main wireless communication technologies used in low-power electronics, such as wearables, beacons, and Internet of Things (IoT) devices. BLE’s energy efficiency characteristic, smart mobiles interoperability, and Over the Air (OTA) capabilities are essential features for ultralow-power devices, which are usually designed with size and cost constraints. Most current research regarding the power analysis of BLE devices focuses on the theoretical aspects of the advertising and scanning cycles, with most results being presented in the form of mathematical models and computer software simulations. Such computer modeling and simulations are important for the comprehension of the technology, but hardware measurement is essential for the understanding of how BLE devices behave in real operation. In addition, recent literature focuses mostly on the BLE technology, leaving possible applications and its analysis out of scope. In this paper, a coin cell battery-powered BLE Data Acquisition Device, with a 4-in-1 sensor and one accelerometer, is proposed and evaluated with respect to its Power Consumption. First, evaluations of the device in advertising mode with the sensors turned off completely, followed by the power analysis when each of the sensors is individually turned on and data is being transmitted, and concluding with the power consumption evaluation when both sensors are on and respectively broadcasting the data to a mobile phone. The results presented in this paper are real-time measurements of the electrical current consumption of the BLE device, where the energy levels that are demonstrated are matched to the BLE behavior and sensor activity.

Keywords: bluetooth low energy, power analysis, BLE advertising cycle, wireless sensor node

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913 Communication Tools Used in Teaching and Their Effects: An Empirical Study on the T. C. Selcuk University Samples

Authors: Sedat Simsek, Tugay Arat

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Today's communication concept, which has a great revolution with the printing press which has been found by Gutenberg, has no boundary thanks to advanced communication devices and the internet. It is possible to take advantage in many areas, such as from medicine to social sciences or from mathematics to education, from the computers that was first produced for the purpose of military services. The use of these developing technologies in the field of education has created a great vision changes in both training and having education. Materials, which can be considered as basic communication resources and used in traditional education has begun to lose its significance, and some technologies have begun to replace them such as internet, computers, smart boards, projection devices and mobile phone. On the other hand, the programs and applications used in these technologies have also been developed. University students use virtual books instead of the traditional printed book, use cell phones instead of note books, use the internet and virtual databases instead of the library to research. They even submit their homework with interactive methods rather than printed materials. The traditional education system, these technologies, which increase productivity, have brought a new dimension to education. The aim of this study is to determine the influence of technologies in the learning process of students and to find whether is there any similarities and differences that arise from the their faculty that they have been educated and and their learning process. In addition to this, it is aimed to determine the level of ICT usage of students studying at the university level. In this context, the advantages and conveniences of the technology used by students are also scrutinized. In this study, we used surveys to collect data. The data were analyzed by using SPSS 16 statistical program with the appropriate testing.

Keywords: education, communication technologies, role of technology, teaching

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912 Iron Response Element-mRNA Binding to Iron Response Protein: Metal Ion Sensing

Authors: Mateen A. Khan, Elizabeth J. Theil, Dixie J. Goss

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Cellular iron homeostasis is accomplished by the coordinated regulated expression of iron uptake, storage, and export. Iron regulate the translation of ferritin and mitochondrial aconitase iron responsive element (IRE)-mRNA by interaction with an iron regulatory protein (IRPs). Iron increases protein biosynthesis encoded in iron responsive element. The noncoding structure IRE-mRNA, approximately 30-nt, folds into a stem loop to control synthesis of proteins in iron trafficking, cell cycling, and nervous system function. Fluorescence anisotropy measurements showed the presence of one binding site on IRP1 for ferritin and mitochondrial aconitase IRE-mRNA. Scatchard analysis revealed the binding affinity (Kₐ) and average binding sites (n) for ferritin and mitochondrial aconitase IRE-mRNA were 68.7 x 10⁶ M⁻¹ and 9.2 x 10⁶ M⁻¹, respectively. In order to understand the relative importance of equilibrium and stability, we further report the contribution of electrostatic interactions in the overall binding of two IRE-mRNA with IRP1. The fluorescence quenching of IRP1 protein was measured at different ionic strengths. The binding affinity of IRE-mRNA to IRP1 decreases with increasing ionic strength, but the number of binding sites was independent of ionic strength. Such results indicate a differential contribution of electrostatics to the interaction of IRE-mRNA with IRP1, possibly related to helix bending or stem interactions and an overall conformational change. Selective destabilization of ferritin and mitochondrial aconitase RNA/protein complexes as reported here explain in part the quantitative differences in signal response to iron in vivo and indicate possible new regulatory interactions.

Keywords: IRE-mRNA, IRP1, binding, ionic strength

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