Search results for: distant cellular interaction
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 4880

Search results for: distant cellular interaction

3350 TiO₂ Deactivation Process during Photocatalytic Ethanol Degradation in the Gas Phase

Authors: W. El-Alami, J. Araña, O. González Díaz, J. M. Doña Rodríguez

Abstract:

The efficiency of the semiconductor TiO₂ needs to be improved to be an effective tool for pollutant removal. To improve the efficiency of this semiconductor, it is necessary to deepen the knowledge of the processes that take place on its surface. In this sense, the deactivation of the catalyst is one of the aspects considered relevant. In order to study this point, the processes of deactivation of TiO₂ during the gas phase degradation of ethanol have been studied. For this, catalysts with only the anatase phase (SA and PC100) and catalysts with anatase and rutile phases (P25 and P90) have been selected. In order to force the deactivation processes, different cycles have been performed, adding ethanol gas but avoiding the degradation of acetates to determine their effect on the process. The surface concentration of fluorine on the catalysts was semi-quantitatively determined by EDAX analysis. The photocatalytic experiments were done with four commercial catalysts (P25, SA, P90, and PC100) and the two fluoride catalysts indicated above. The interaction and photocatalytic degradation of ethanol were followed by Fourier transform infrared spectroscopy (FTIR). EDAX analysis has revealed the presence of sodium on the surface of fluorinated catalysts. In FTIR studies, it has been observed that the acetates adsorbed on the anatase phase in P25 and P90 give rise to electron transfer to surface traps that modify the electronic states of the semiconductor. These deactivation studies have also been carried out with fluorinated P25 and SA catalysts (F-P25 and F-SA) which have observed similar electron transfers but in the opposite direction during illumination. In these materials, it has been observed that the electrons present in the surface traps, as a consequence of the interaction Ti-F, react with the holes, causing a change in the electronic states of the semiconductor. In this way, deactivated states of these materials have been detected by different electron transfer routes. It has been identified that acetates produced from the degradation of ethanol in P25 and P90 are probably hydrated on the surface of the rutile phase. In the catalysts with only the anatase phase (SA and PC100), the deactivation is immediate if the acetates are not removed before adsorbing ethanol again. In F-P25 and F-SA has been observed that the acetates formed react with the sodium ions present on the surface and not with the Ti atoms because they are interacting with the fluorine.

Keywords: photocatalytic degradation, ethanol, TiO₂, deactivation process, F-P25

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3349 Experimental and Theoretical Characterization of Supramolecular Complexes between 7-(Diethylamino)Quinoline-2(1H)-One and Cucurbit[7] Uril

Authors: Kevin A. Droguett, Edwin G. Pérez, Denis Fuentealba, Margarita E. Aliaga, Angélica M. Fierro

Abstract:

Supramolecular chemistry is a field of growing interest. Moreover, studying the formation of host-guest complexes between macrocycles and dyes is highly attractive due to their potential applications. Examples of the above are drug delivery, catalytic process, and sensing, among others. There are different dyes of interest in the literature; one example is the quinolinone derivatives. Those molecules have good optical properties and chemical and thermal stability, making them suitable for developing fluorescent probes. Secondly, several macrocycles can be seen in the literature. One example is the cucurbiturils. This water-soluble macromolecule family has a hydrophobic cavity and two identical carbonyl portals. Additionally, the thermodynamic analysis of those supramolecular systems could help understand the affinity between the host and guest, their interaction, and the main stabilization energy of the complex. In this work, two 7-(diethylamino) quinoline-2 (1H)-one derivative (QD1-2) and their interaction with cucurbit[7]uril (CB[7]) were studied from an experimental and in-silico point of view. For the experimental section, the complexes showed a 1:1 stoichiometry by HRMS-ESI and isothermal titration calorimetry (ITC). The inclusion of the derivatives on the macrocycle lends to an upward shift in the fluorescence intensity, and the pKa value of QD1-2 exhibits almost no variation after the formation of the complex. The thermodynamics of the inclusion complexes was investigated using ITC; the results demonstrate a non-classical hydrophobic effect with a minimum contribution from the entropy term and a constant binding on the order of 106 for both ligands. Additionally, dynamic molecular studies were carried out during 300 ns in an explicit solvent at NTP conditions. Our finding shows that the complex remains stable during the simulation (RMSD ~1 Å), and hydrogen bonds contribute to the stabilization of the systems. Finally, thermodynamic parameters from MMPBSA calculations were obtained to generate new computational insights to compare with experimental results.

Keywords: host-guest complexes, molecular dynamics, quinolin-2(1H)-one derivatives dyes, thermodynamics

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3348 The 6Rs of Radiobiology in Photodynamic Therapy: Review

Authors: Kave Moloudi, Heidi Abrahamse, Blassan P. George

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Radiotherapy (RT) and photodynamic therapy (PDT) are both forms of cancer treatment that aim to kill cancer cells while minimizing damage to healthy tissue. The similarity between RT and PDT lies in their mechanism of action. Both treatments use energy to damage cancer cells. RT uses high-energy radiation to damage the DNA of cancer cells, while PDT uses light energy to activate a photosensitizing agent, which produces reactive oxygen species (ROS) that damage the cancer cells. Both treatments require careful planning and monitoring to ensure the correct dose is delivered to the tumor while minimizing damage to surrounding healthy tissue. They are also often used in combination with other treatments, such as surgery or chemotherapy, to improve overall outcomes. However, there are also significant differences between RT and PDT. For example, RT is a non-invasive treatment that can be delivered externally or internally, while PDT requires the injection of a photosensitizing agent and the use of a specialized light source to activate it. Additionally, the side effects and risks associated with each treatment can vary. In this review, we focus on generalizing the 6Rs of radiobiology in PDT, which can open a window for the clinical application of Radio-photodynamic therapy with minimum side effects. Furthermore, this review can open new insight to work on and design new radio-photosensitizer agents in Radio-photodynamic therapy.

Keywords: radiobiology, photodynamic therapy, radiotherapy, 6Rs in radiobiology, ROS, DNA damages, cellular and molecular mechanism, clinical application.

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3347 Trastuzumab Decorated Bioadhesive Nanoparticles for Targeted Breast Cancer Therapy

Authors: Kasi Viswanadh Matte, Abhisheh Kumar Mehata, M.S. Muthu

Abstract:

Brest cancer, up-regulated with human epidermal growth factor receptor type-2 (HER-2) led to the concept of developing HER-2 targeted anticancer therapeutics. Docetaxel-loaded D-α-tocopherol polyethylene glycol succinate 1000 conjugated chitosan (TPGS-g-chitosan) nanoparticles were prepared with or without Trastuzumab decoration. The particle size and entrapment efficiency of conventional, non-targeted and targeted nanoparticles were found to be in the range of 126-186 nm and 74-78% respectively. In-vitro, MDA-MB-231 cells showed that docetaxel-loaded non-targeted and HER-2 receptor targeted TPGS-g-chitosan nanoparticles have enhanced the cellular uptake and cytotoxicity with a promising bioadhesion property, in comparison to conventional nanoparticles. The IC50 values of non-targeted and targeted nanoparticles from cytotoxic assay were found to be 43 and 223 folds higher than DocelTM. The in-vivo pharmacokinetic study showed 2.33, and 2.82-fold enhancement in relative bioavailability of docetaxel for non-targeted and HER-2 receptor targeted nanoparticles, respectively than DocelTM, and after i.v administration, non-targeted and targeted nanoparticle achieved 3.48 and 5.94 times prolonged half-life in comparison to DocelTM. The area under the curve (AUC), relative bioavailability (FR) and mean residence time (MRT) were found to be higher for non-targeted and targeted nanoparticles compared to DocelTM. Further, histopathology results of non-targeted and targeted nanoparticles showed less toxicity on vital organs such as lungs, liver, and kidney compared to DocelTM.

Keywords: breast cancer, HER-2 receptor, targeted nanomedicine, chitosan, TPGS

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3346 Gender Gap in Education and Empowerment Influenced by Parents’ Attitude

Authors: N. Kashif, M. K. Naseer

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This is an undeniable fact that parents are the very first role model for their children and children are the silent observers and followers of their parents. The environment they would be provided will leave either positive or negative lasting impact on their physical and mental behavior and abilities to grow, progress and conquer. This paper focuses on the observation particularly in South Asian countries where females have been facing problems in accessing education and getting financially independent or stable. This paper emphasizes on a survey conducted in rural areas of Punjab State in Pakistan. It explains how the parents’ educational background, financial status, conservative and interdependent accommodation style influence a prominent inequality of giving their female child right to study and get empowered. The forces behind this gender discrimination are not limited to parents’ life style impact but also include some major social problems like distant schools, gender-based harassment, and threat, insecurities, employment opportunities, so on. As a grass root level solution, it is proposed to develop an institution which collects data regarding child birth in their region and can contact the parent when their child is ready to start school. Building up trust based relationship with parents is the most crucial and significant factor. Secondly, celebrities and public figures can play an extraordinary role in running a campaign to advocate and encourage people living in rural areas, villages and small towns. All possible solutions can never be implemented without the support of the state government. Therefore, this paper invites more thoughtful actions, properly planned strategies, initiators to take the lead and make a platform for those who are underprivileged and deprived of their basic rights. Any country, where female constitute 49% of its entire population can never progress without promoting female empowerment and their right to compulsory education, and it is never late or impossible to admit the facts and practically start a flexible solution- oriented approach.

Keywords: employment opportunities, female empowerment, gender based harassment, gender discrimination, inequality, parents' life style impact

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3345 Safety and Efficacy of Recombinant Clostridium botulinum Types B Vaccine Candidate

Authors: Mi-Hye Hwang, Young Min Son, Kichan Lee, Bang-Hun Hyun, Byeong Yeal Jung

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Botulism is a paralytic disease of human beings and animals caused by neurotoxin produced by Clostridium botulinum. The neurotoxins are genetically distinguished into 8 types, A to H. Ingestion of performed toxin, usually types B, C, and D, have been shown to produce diseases in most cases of cattle botulism. Vaccination is the best measure to prevent cattle botulism. However, the commercially available toxoid-based vaccines are difficult and hazardous to produce. We produced recombinant protein using gene of heavy chain domain of botulinum toxin B of which binds to cellular receptor of neuron cells and used as immunogen. In this study, we evaluated the safety and efficacy of botulism vaccine composed of recombinant types B. Safety test was done by National Regulation for Veterinary Biologicals. For efficacy test, female ICR mice (5 weeks old) were subcutaneously injected, intraperitoneally challenged, and examined the survival rates compared with vaccination and non-vaccination group. Mouse survival rate of recombinant types B vaccine was above 80%, while one of non-vaccination group was 0%. A vaccine composed of recombinant types B was safe and efficacious in mouse. Our results suggest that recombinant heavy chain receptor binding domain can be used as an effective vaccine candidate for type B botulism.

Keywords: botulism, livestock, vaccine, recombinant protein, toxin

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3344 Evidence of Half-Metallicity in Cubic PrMnO3 Perovskite

Authors: B. Bouadjemi, S. Bentata, W. Benstaali, A. Abbad

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The electronic and magnetic properties of the cubic praseodymium oxides perovskites PrMnO3 were calculated using the density functional theory (DFT) with both generalized gradient approximation (GGA) and GGA+U approaches, where U is on-site Coulomb interaction correction. The results show a half-metallic ferromagnetic ground state for PrMnO3 in GGA+U approached, while semi-metallic ferromagnetic character is observed in GGA. The results obtained, make the cubic PrMnO3 a promising candidate for application in spintronics.

Keywords: first-principles, electronic properties, transition metal, materials science

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3343 Histological Evaluation of the Neuroprotective Roles of Trans Cinnamaldehyde against High Fat Diet and Streptozotozin Induced Neurodegeneration in Wistar Rats

Authors: Samson Ehindero, Oluwole Akinola

Abstract:

Substantial evidence has shown an association between type 2 diabetes (T2D) and cognitive decline, Trans Cinnamaldehyde (TCA) has been shown to have many potent pharmacological properties. In this present study, we are currently investigating the effects of TCA on type II diabetes-induced neurodegeneration. Neurodegeneration was induced in forty (40) adult wistar rats using high fat diet (HFD) for 4 months followed by low dose of streptozotocin (STZ) (40 mg/kg, i.p.) administration. TCA was administered orally for 30 days at the doses of 40mg/kg and 60mg/kg body weight. Animals were randomized and divided into following groups; A- control group, B- diabetic group, C- TCA (high dose), D- diabetic + TCA (high dose), E- diabetic + TCA (high dose) with high fat diet, F- TCA Low dose, G- diabetic + TCA (low dose) and H- diabetic + TCA (low dose) with high fat diet. Animals were subjected to behavioral tests followed by histological studies of the hippocampus. Demented rats showed impaired behavior in Y- Maze test compared to treated and control groups. Trans Cinnamaldehyde restores the histo architecture of the hippocampus of demented rats. This present study demonstrates that treatment with trans- cinnamaldehyde improves behavioral deficits, restores cellular histo architecture in rat models of neurodegeneration.

Keywords: neurodegeneration, trans cinnamaldehyde, high fat diet, streptozotocin

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3342 Microfabrication and Non-Invasive Imaging of Porous Osteogenic Structures Using Laser-Assisted Technologies

Authors: Irina Alexandra Paun, Mona Mihailescu, Marian Zamfirescu, Catalin Romeo Luculescu, Adriana Maria Acasandrei, Cosmin Catalin Mustaciosu, Roxana Cristina Popescu, Maria Dinescu

Abstract:

A major concern in bone tissue engineering is to develop complex 3D architectures that mimic the natural cells environment, facilitate the cells growth in a defined manner and allow the flow transport of nutrients and metabolic waste. In particular, porous structures of controlled pore size and positioning are indispensable for growing human-like bone structures. Another concern is to monitor both the structures and the seeded cells with high spatial resolution and without interfering with the cells natural environment. The present approach relies on laser-based technologies employed for fabricating porous biomimetic structures that support the growth of osteoblast-like cells and for their non-invasive 3D imaging. Specifically, the porous structures were built by two photon polymerization –direct writing (2PP_DW) of the commercially available photoresists IL-L780, using the Photonic Professional 3D lithography system. The structures consist of vertical tubes with micrometer-sized heights and diameters, in a honeycomb-like spatial arrangement. These were fabricated by irradiating the IP-L780 photoresist with focused laser pulses with wavelength centered at 780 nm, 120 fs pulse duration and 80 MHz repetition rate. The samples were precisely scanned in 3D by piezo stages. The coarse positioning was done by XY motorized stages. The scanning path was programmed through a writing language (GWL) script developed by Nanoscribe. Following laser irradiation, the unexposed regions of the photoresist were washed out by immersing the samples in the Propylene Glycol Monomethyl Ether Acetate (PGMEA). The porous structures were seeded with osteoblast like MG-63 cells and their osteogenic potential was tested in vitro. The cell-seeded structures were analyzed in 3D using the digital holographic microscopy technique (DHM). DHM is a marker free and high spatial resolution imaging tool, where the hologram acquisition is performed non-invasively i.e. without interfering with the cells natural environment. Following hologram recording, a digital algorithm provided a 3D image of the sample, as well as information about its refractive index, which is correlated with the intracellular content. The axial resolution of the images went down to the nanoscale, while the temporal scales ranged from milliseconds up to hours. The hologram did not involve sample scanning and the whole image was available in one frame recorded going over 200μm field of view. The digital holograms processing provided 3D quantitative information on the porous structures and allowed a quantitative analysis of the cellular response in respect to the porous architectures. The cellular shape and dimensions were found to be influenced by the underlying micro relief. Furthermore, the intracellular content gave evidence on the beneficial role of the porous structures in promoting osteoblast differentiation. In all, the proposed laser-based protocol emerges as a promising tool for the fabrication and non-invasive imaging of porous constructs for bone tissue engineering. Acknowledgments: This work was supported by a grant of the Romanian Authority for Scientific Research and Innovation, CNCS-UEFISCDI, project PN-II-RU-TE-2014-4-2534 (contract 97 from 01/10/2015) and by UEFISCDI PN-II-PT-PCCA no. 6/2012. A part of this work was performed in the CETAL laser facility, supported by the National Program PN 16 47 - LAPLAS IV.

Keywords: biomimetic, holography, laser, osteoblast, two photon polymerization

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3341 Pegylated Liposomes of Trans Resveratrol, an Anticancer Agent, for Enhancing Therapeutic Efficacy and Long Circulation

Authors: M. R. Vijayakumar, Sanjay Kumar Singh, Lakshmi, Hithesh Dewangan, Sanjay Singh

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Trans resveratrol (RES) is a natural molecule proved for cancer preventive and therapeutic activities devoid of any potential side effects. However, the therapeutic application of RES in disease management is limited because of its rapid elimination from blood circulation thereby low biological half life in mammals. Therefore, the main objective of this study is to enhance the circulation as well as therapeutic efficacy using PEGylated liposomes. D-α-tocopheryl polyethylene glycol 1000 succinate (vitamin E TPGS) is applied as steric surface decorating agent to prepare RES liposomes by thin film hydration method. The prepared nanoparticles were evaluated by various state of the art techniques such as dynamic light scattering (DLS) technique for particle size and zeta potential, TEM for shape, differential scanning calorimetry (DSC) for interaction analysis and XRD for crystalline changes of drug. Encapsulation efficiency and invitro drug release were determined by dialysis bag method. Cancer cell viability studies were performed by MTT assay, respectively. Pharmacokinetic studies were performed in sprague dawley rats. The prepared liposomes were found to be spherical in shape. Particle size and zeta potential of prepared formulations varied from 64.5±3.16 to 262.3±7.45 nm and -2.1 to 1.76 mV, respectively. DSC study revealed absence of potential interaction. XRD study revealed presence of amorphous form in liposomes. Entrapment efficiency was found to be 87.45±2.14 % and the drug release was found to be controlled up to 24 hours. Minimized MEC in MTT assay and tremendous enhancement in circulation time of RES PEGylated liposomes than its pristine form revealed that the stearic stabilized PEGylated liposomes can be an alternative tool to commercialize this molecule for chemopreventive and therapeutic applications in cancer.

Keywords: trans resveratrol, cancer nanotechnology, long circulating liposomes, bioavailability enhancement, liposomes for cancer therapy, PEGylated liposomes

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3340 Use of Social Media in Political Communications: Example of Facebook

Authors: Havva Nur Tarakci, Bahar Urhan Torun

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The transformation that is seen in every area of life by technology, especially internet technology changes the structure of political communications too. Internet, which is at the top of new communication technologies, affects political communications with its structure in a way that no traditional communication tools ever have and enables interaction and the channel between receiver and sender, and it becomes one of the most effective tools preferred among the political communication applications. This state as a result of technological convergence makes Internet an unobtainable place for political communication campaigns. Political communications, which means every kind of communication strategies that political parties called 'actors of political communications' use with the aim of messaging their opinions and party programmes to their present and potential voters who are a target group for them, is a type of communication that is frequently used also among social media tools at the present day. The electorate consisting of different structures is informed, directed, and managed by social media tools. Political parties easily reach their electorate by these tools without any limitations of both time and place and also are able to take the opinions and reactions of their electorate by the element of interaction that is a feature of social media. In this context, Facebook, which is a place that political parties use in social media at most, is a communication network including in our daily life since 2004. As it is one of the most popular social networks today, it is among the most-visited websites in the global scale. In this way, the research is based on the question, “How do the political parties use Facebook at the campaigns, which they conduct during the election periods, for informing their voters?” and it aims at clarifying the Facebook using practices of the political parties. In direction of this objective the official Facebook accounts of the four political parties (JDP–AKParti, PDP–BDP, RPP-CHP, NMP-MHP), which reach their voters by social media besides other communication tools, are treated, and a frame for the politics of Turkey is formed. The time of examination is constricted with totally two weeks, one week before the mayoral elections and one week after the mayoral elections, when it is supposed that the political parties use their Facebook accounts in full swing. As a research method, the method of content analysis is preferred, and the texts and the visual elements that are gotten are interpreted based on this analysis.

Keywords: Facebook, political communications, social media, electrorate

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3339 The Effects of Inoculation and N Fertilization on Soybean (Glycine max (L.) Merr.) Seed Yield and Protein Concentration under Drought Stress

Authors: Oqba Basal, Andras Szabo

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Using mineral fertilization is increasing worldwide, as it is claimed to be majorly responsible for achieving high yields; however, the negative impacts of mineral fertilization on soil and environment are becoming more obvious, with alternative methods being more necessary and applicable, especially with the current climatic changes which have imposed serious abiotic stresses, such as drought. An experiment was made during 2017 growing season in Debrecen, Hungary to investigate the effects of inoculation and N fertilization on the seed yield and protein concentration of the soybean (Glycine max (L.) Merr.) cultivar (Panonia Kincse) under three different irrigation regimes: severe drought stress (SD), moderate drought stress (MD) and control with no drought stress (ND). Three N fertilizer rates were applied: no N fertilizer (0 N), 35 kg ha⁻¹ of N fertilizer (35 N) and 105 kg ha⁻¹ of N fertilizer (105 N). Half of the seeds in each treatment was inoculated with Bradyrhizobium japonicum inoculant, and the other half was not inoculated. The results showed significant differences in the seed yield associated with inoculation, irrigation and the interaction between them, whereas there were no significant differences in the seed yield associated with fertilization alone or in interaction with inoculation or irrigation or both. When seeds were inoculated, yield was increased when (35 N) was applied compared to (0 N) but not significantly; however, the high rate of N fertilizer (105 N) reduced the yield to a level even less than (0 N). When seeds were not inoculated, the highest rate of N increased the yield the most compared to the other two N fertilizer rates whenever the drought was present (moderate or severe). Under severe drought stress, inoculation was positively and significantly correlated with yield; however, adding N fertilizer increased the yield of uninoculated plants compared to the inoculated ones, regardless of the rate of N fertilizer. Protein concentration in the seeds was significantly affected by irrigation and by fertilization, but not by inoculation. Protein concentration increased as the N fertilization rate increased, regardless of the inoculation or irrigation treatments; moreover, increasing the N rate reduced the correlation coefficient of protein concentration with the irrigation. It was concluded that adding N fertilizer is not always recommended, especially when seeds are inoculated before being sown; however, it is very important under severe drought stress to sustain yield. Enhanced protein concentrations could be achieved by applying N fertilization, whether the seeds were pre-inoculated or not.

Keywords: drought stress, N fertilization, protein concentration, soybean

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3338 Synthesis and Applications of Biosorbent from Barley Husk for Adsorption of Heavy Metals and Bacteria from Water

Authors: Sudarshan Kalsulkar, Sunil S. Bhagwat

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Biosorption is a physiochemical process that occurs naturally in certain biomass which allows it to passively concentrate and bind contaminants onto its cellular structure. Activated carbons (AC) are one such efficient biosorbents made by utilizing lignocellulosic materials from agricultural waste. Steam activated carbon (AC) was synthesized from Barley husk. Its synthesis parameters of time and temperature were optimized. Its physico-chemical properties like density, surface area, pore volume, Methylene blue and Iodine values were characterized. BET surface area was found to be 42 m²/g. Batch Adsorption tests were carried out to determine the maximum adsorption capacity (qmax) for various metal ions. Cd+2 48.74 mg/g, Pb+2 19.28 mg/g, Hg+2 39.1mg/g were the respective qmax values. pH and time were optimized for adsorption of each ion. Column Adsorptions were carried for each to obtain breakthrough data. Microbial adsorption was carried using E. coli K12 strain. 78% reduction in cell count was observed at operating conditions. Thus the synthesized Barley husk AC can be an economically feasible replacement for commercially available AC prepared from the costlier coconut shells. Breweries and malting industries where barley husk is a primary waste generated on a large scale can be a good source for bulk raw material.

Keywords: activated carbon, Barley husk, biosorption, decontamination, heavy metal removal, water treatment

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3337 Investigation of Performance of Organic Acids on Carbonate Rocks (Experimental Study in Ahwaz Oilfield)

Authors: Azad Jarrahian, Ehsan Heidaryan

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Matrix acidizing treatments can yield impressive production increase if properly applied. In this study, carbonate samples taken from Ahwaz Oilfield have undergone static solubility, sludge, emulsion, and core flooding tests. In each test interaction of acid and rock is reported and at the end it has been shown that how initial permeability and type of acid affects the overall treatment efficiency.

Keywords: carbonate acidizing, organic acids, spending rate, acid penetration, incomplete spending.

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3336 Chitosan Modified Halloysite Nanomaterials for Efficient and Effective Vaccine Delivery in Farmed Fish

Authors: Saji George, Eng Khuan Seng, Christof Luda

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Nanotechnology has been recognized as an important tool for modern agriculture and has the potential to overcome some of the pressing challenges faced by aquaculture industry. A strategy for optimizing nanotechnology-based therapeutic delivery platform for immunizing farmed fish was developed. Accordingly, a compositional library of nanomaterials of natural chemistry (Halloysite (clay), Chitosan, Hydroxyapatite, Mesoporous Silica and a composite material of clay-chitosan) was screened for their toxicity and efficiency in delivering models antigens in cellular and zebrafish embryo models using high throughput screening platforms. Through multi-parametric optimization, chitosan modified halloysite (clay) nanomaterial was identified as an optimal vaccine delivery platform. Further, studies conducted in juvenile seabass showed the potential of clay-chitosan in delivering outer membrane protein of Tenacibaculum maritimum- TIMA (pathogenic bacteria) to and its efficiency in eliciting immune responses in fish. In short, as exemplified by this work, the strategy of using compositional nanomaterial libraries and their biological profiling using high-throughput screening platform could fasten the discovery process of nanomaterials with potential applications in food and agriculture.

Keywords: nanotechnology, fish-vaccine, drug-delivery, halloysite-chitosan

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3335 Gender Differences in Objectively Assessed Physical Activity among Urban 15-Year-Olds

Authors: Marjeta Misigoj Durakovic, Maroje Soric, Lovro Stefan

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Background and aim: Physical inactivity has been linked with increased morbidity and premature mortality and adolescence has been recognised as the critical period for a decline in physical activity (PA) level. In order to properly direct interventions aimed at increasing PA, high-risk groups of individuals should be identified. Therefore, the aim of this study is to describe gender differences in: a) PA level; b) weekly PA patterns. Methods: This investigation is a part of the CRO-PALS study which is an on-going longitudinal study conducted in a representative sample of urban youth in Zagreb (Croatia). CRO-PALS involves 903 adolescents and for the purpose of this study data from a subgroup of 190 participants with information on objective PA level were analysed (116 girls; mean age [SD]=15.6[0.3] years). Duration of moderate and vigorous PA was measured during 5 consecutive by a multiple-sensor physical activity monitor (SenseWear Armband, BodyMedia inc., Pittsburgh, USA). Gender differences in PA level were evaluated using independent samples t-test. Differences in school week and weekend levels of activity were assessed using mixed ANOVA with gender as between-subjects factor. The amount of vigorous PA had to be log-transformed to achieve normality in the distribution. Results: Boys were more active than girls. Duration of moderate-to-vigorous PA averaged 111±44 min/day in boys and 80±38 min/day in girls (mean difference=31 min/day, 95%CI=20-43 min/day). Vigorous PA was 2.5 times higher in boys compared to girls (95%CI=1.9-3.5). Participants were more active during school days than on weekends. The magnitude of the difference in moderate-to-vigorous PA was similar in both gender (p value for time*gender interaction = 0.79) and averaged 19 min/day (95%CI=11-27 min/day). Similarly, vigorous PA was 36% lower on weekends compared with school days (95%CI=22-46%) with no gender difference (p value for time*gender interaction = 0.52). Conclusion: PA level was higher in boys than in girls throughout the week. Still, in both boys and girls, the amount of PA reduced markedly on weekends compared with school days.

Keywords: adolescence, multiple-sensor physical activity monitor, physical activity level, weekly physical activity pattern

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3334 Investigating the Role of Dystrophin in Neuronal Homeostasis

Authors: Samantha Shallop, Hakinya Karra, Tytus Bernas, Gladys Shaw, Gretchen Neigh, Jeffrey Dupree, Mathula Thangarajh

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Abnormal neuronal homeostasis is considered a structural correlate of cognitive deficits in Duchenne Muscular Dystrophy. Neurons are highly polarized cells with multiple dendrites but a single axon. Trafficking of cellular organelles are highly regulated, with the cargo in the somatodendritic region of the neuron not permitted to enter the axonal compartment. We investigated the molecular mechanisms that regular organelle trafficking in neurons using a multimodal approach, including high-resolution structural illumination, proteomics, immunohistochemistry, and computational modeling. We investigated the expression of ankyrin-G, the master regulator controlling neuronal polarity. The expression of ankyrin G and the morphology of the axon initial segment was profoundly abnormal in the CA1 hippocampal neurons in the mdx52 animal model of DMD. Ankyrin-G colocalized with kinesin KIF5a, the anterograde protein transporter, with higher levels in older mdx52 mice than younger mdx52 mice. These results suggest that the functional trafficking from the somatodendritic compartment is abnormal. Our data suggests that dystrophin deficiency compromised neuronal homeostasis via ankyrin-G-based mechanisms.

Keywords: neurons, axonal transport, duchenne muscular dystrophy, organelle transport

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3333 Sexual Consent: Exploring the Perceptions of Heterosexual, Gay, and Bisexual Men

Authors: Shulamit Sternin, Raymond M. McKie, Carter Winberg, Robb N. Travers, Terry P. Humphreys, Elke D. Reissing

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Issues surrounding sexual consent negotiation have become a major topic of societal concern. The majority of current research focuses on the complexities of sexual consent negotiations and the multitude of nuanced issues that surround the consent obtainment of heterosexual adults in post-secondary educational institutions. To date, the only study that has addressed sexual consent negotiation behaviour in same-sex relationships focused on the extent to which individuals used a variety of different verbal and nonverbal sexual consent behaviours to initiate or respond to sexual activity. The results were consistent with trends found within heterosexual individuals; thus, suggesting that the current understanding of sexual consent negotiation, which is grounded in heterosexual research, can serve as a strong foundation for further exploration of sexual consent negotiation within same-sex relationships populations. The current study quantitatively investigated the differences between heterosexual men and gay and bisexual men (GBM) in their understanding of sexual consent negotiation. Exploring how the perceptions of GBM differ from heterosexual males provides insight into some of the unique challenges faced by GBM. Data were collected from a sample of 252 heterosexual men and 314 GBM from Canada, the United States, and Western Europe. Participants responded to the question, 'do you think sexual consent and sex negotiation is different for heterosexual men compared to gay men? If so, how?' by completed an online survey. Responses were analysed following Braun & Clarke’s (2006) six phase thematic analysis guidelines. Inter-rater coding was validated using Cohen’s Kappa value and was calculated at (ϰ = 0.84), indicating a very strong level of agreement between raters. The final thematic structure yielded four major themes: understanding of sexual interaction, unique challenges, scripted role, and universal consent. Respondents spoke to their understanding of sexual interaction, believing GBM sexual consent negotiation to be faster and more immediate. This was linked to perceptions of emotional attachment and the idea that sexual interaction and emotional involvement were distinct and separate processes in GBM sexual consent negotiation, not believed to be the case in heterosexual interactions. Unique challenges such as different protection concerns, role declaration, and sexualization of spaces were understood to hold differing levels of consideration for heterosexual men and GBM. The perception of a clearly defined sexual script for GBM was suggested as a factor that may create ambiguity surrounding sexual consent negotiation, which in turn holds significant implications on unwanted sexual experiences for GBM. Broadening the scope of the current understanding of sexual consent negotiation by focusing on heterosexual and GBM population, the current study has revealed variations in perception of sexual consent negotiation between these two populations. These differences may be understood within the context of sexual scripting theory and masculinity gender role theory. We suggest that sexual consent negotiation is a health risk factor for GBM that has not yet been adequately understood and addressed. Awareness of the perceptions that surround the sexual consent negotiation of both GBM and heterosexual men holds implications on public knowledge, which in turn can better inform policy making, education, future research, and clinical treatment.

Keywords: sexual consent, negotiation, heterosexual men, GBM, sexual script

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3332 Diagnosis of Gingivitis Based on Correlations of Laser Doppler Data and Gingival Fluid Cytology

Authors: A. V. Belousov, Yakushenko

Abstract:

One of the main problems of modern dentistry is development a reliable method to detect inflammation in the gums on the stages of diagnosis and assessment of treatment efficacy. We have proposed a method of gingival fluid intake, which successfully combines accessibility, excluding the impact of the annoying and damaging the gingival sulcus factors and provides reliable results (patent of RF№ 2342956 Method of gingival fluid intake). The objects of the study were students - volunteers of Dentistry Faculty numbering 75 people aged 20-21 years. Cellular composition of gingival fluid was studied using microscope "Olympus CX 31" (Japan) with the calculation of epithelial leukocyte index (ELI). Assessment of gingival micro circulation was performed using the apparatus «LAKK–01» (Lazma, Moscow). Cytological investigation noted the highly informative of epithelial leukocyte index (ELI), which demonstrated changes in the mechanisms of protection gums. The increase of ELI occurs during inhibition mechanisms of phagocytosis and activation of epithelial desquamation. The cytological data correlate with micro circulation indicators obtained by laser Doppler flowmetry. We have identified and confirmed the correlations between parameters laser Doppler flowmetry and data cytology gingival fluid in patients with gingivitis.

Keywords: gingivitis, laser doppler flowmetry, gingival fluid cytology, epithelial leukocyte index (ELI)

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3331 Human-Automation Interaction in Law: Mapping Legal Decisions and Judgments, Cognitive Processes, and Automation Levels

Authors: Dovile Petkeviciute-Barysiene

Abstract:

Legal technologies not only create new ways for accessing and providing legal services but also transform the role of legal practitioners. Both lawyers and users of legal services expect automated solutions to outperform people with objectivity and impartiality. Although fairness of the automated decisions is crucial, research on assessing various characteristics of automated processes related to the perceived fairness has only begun. One of the major obstacles to this research is the lack of comprehensive understanding of what legal actions are automated and could be meaningfully automated, and to what extent. Neither public nor legal practitioners oftentimes cannot envision technological input due to the lack of general without illustrative examples. The aim of this study is to map decision making stages and automation levels which are and/or could be achieved in legal actions related to pre-trial and trial processes. Major legal decisions and judgments are identified during the consultations with legal practitioners. The dual-process model of information processing is used to describe cognitive processes taking place while making legal decisions and judgments during pre-trial and trial action. Some of the existing legal technologies are incorporated into the analysis as well. Several published automation level taxonomies are considered because none of them fit well into the legal context, as they were all created for avionics, teleoperation, unmanned aerial vehicles, etc. From the information processing perspective, analysis of the legal decisions and judgments expose situations that are most sensitive to cognitive bias, among others, also help to identify areas that would benefit from the automation the most. Automation level analysis, in turn, provides a systematic approach to interaction and cooperation between humans and algorithms. Moreover, an integrated map of legal decisions and judgments, information processing characteristics, and automation levels all together provide some groundwork for the research of legal technology perceived fairness and acceptance. Acknowledgment: This project has received funding from European Social Fund (project No 09.3.3-LMT-K-712-19-0116) under grant agreement with the Research Council of Lithuania (LMTLT).

Keywords: automation levels, information processing, legal judgment and decision making, legal technology

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3330 Using Eye-Tracking Technology to Understand Consumers’ Comprehension of Multimedia Health Information

Authors: Samiullah Paracha, Sania Jehanzeb, M. H. Gharanai, A. R. Ahmadi, H.Sokout, Toshiro Takahara

Abstract:

The purpose of this study is to examine how health consumers utilize pictures when developing an understanding of multimedia health documents, and whether attentional processes, measured by eye-tracking, relate to differences in health-related cognitive resources and passage comprehension. To investigate these issues, we will present health-related text-picture passages to elders and collect eye movement data to measure readers’ looking behaviors.

Keywords: multimedia, eye-tracking, consumer health informatics, human-computer interaction

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3329 The Study of the Absorption and Translocation of Chromium by Lygeum spartum in the Mining Region of Djebel Hamimat and Soil-Plant Interaction

Authors: H. Khomri, A. Bentellis

Abstract:

Since century of the Development Activities extraction and a dispersed mineral processing Toxic metals and much more contaminated vast areas occupied by what they natural outcrops. New types of metalliferous habitats are so appeared. A species that is Lygeum spartum attracted our curiosity because apart from its valuable role in desertification, it is apparently able to exclude antimony and other metals can be. This species, green leaf blades which are provided as cattle feed, would be a good subject for phytoremediation of mineral soils. The study of absorption and translocation of chromium by the Lygeum spartum in the mining region of Djebel Hamimat and the interaction soil-plant, revealed that soils of this species living in this region are alkaline, calcareous majority in their fine texture medium and saline in their minority. They have normal levels of organic matter. They are moderately rich in nitrogen. They contain total chromium content reaches a maximum of 66,80 mg Kg^(-1) and a total absence of soluble chromium. The results of the analysis of variance of the difference between bare soils and soils appear Lygeum spartum made a significant difference only for the silt and organic matter. But for the other variables analyzed this difference is not significant. Thus, this plant has only one action on the amendment, only the levels of silt and organic matter in soils. The results of the multiple regression of the chromium content of the roots according to all soil variables studied did appear that among the studied variables included in the model, only the electrical conductivity and clay occur in the explanation of contents chromium in roots. The chromium content of the aerial parts analyzed by regression based on all studied soil variables allows us to see only the variables: electrical conductivity and content of chromium in the root portion involved in the explanation of the content chromium in the aerial part.

Keywords: absorption, translocation, analysis of variance, chrome, Lygeum spartum, multiple regression, the soil variables

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3328 Role of F18-FDG PET in Management of Differentiated Thyroid Cancers (TENIS) Patients

Authors: Seemab Safdar, Shazia Fatima, Ahmad Qureshy, M. Adnan Saeed, M. Faheem

Abstract:

Background: Thyroid cancer has 586,000 cases per year worldwide, and this translates to 3% of all tumor diagnoses. 90% of the cases fall under differentiated thyroid carcinoma (DTC), which includes follicular thyroid cancer (FTC) and papillary thyroid cancer (PTC). During their illness, 10% of patients develop distant metastases, and two-thirds of them develop resistance to radioactive iodine (RAI) treatment. It has been shown that in some circumstances, like DTC with high TG levels and negative 131I whole-body scintigraphy (TENIS), [18F] FDG-PET-CT is an effective imaging technique. This study determines the role of [18F] FDG-PET-CT in the treatment of TENIS patients. Methods: 16 patients (n = 12 female; 4 males, age 45 ± 15 years) with histologically proven thyroid cancer (Differentiated and poorly differentiated) and high TG with negative iodine scans were included in this prospective study from January 2024 to June 2024. They underwent scanning in state-of-the-art (GE Discovery MI) [18F] FDG-PET-CT for re-staging or diagnostics of recurrent disease using a standardized protocol. All DTC subtypes and PDTC were included. The referring physicians completed standardized questionnaires both before and after PET-CT to prospectively determine the examination's effect on clinical decision-making. Patient outcomes were measured by analysis of medical records. Moreover, after PET-CT, a change in the pre-PET-CT planned therapies was documented in 32% of cases and additional invasive diagnostic procedures could be waived in 37.5 % of cases. TG levels under TSH stimulation were significantly higher in patients showing PET-CT metastases compared to patients without such findings (68.75%). Results: Without PET-CT, physicians referring to the doctors had not established a complete treatment plan for 45% of patients with thyroid carcinoma. 12/16 patients showed FDG avidity in cervical lymph nodes that were not Iodine avid previously, 2 patients had FDG avid disease in the lungs. In the process, PET-CT helped plan patient management and created a clear plan for treatment in 68.75% of patients. Conclusions: This study confirms that [18F] FDG-PET-CT used in a routine clinical setting has a very important impact on the management of patients with thyroid cancer when TG levels are persistently high in the presence of negative Iodine Scans by initiating treatments and replacing additional imaging and invasive tests.

Keywords: PET-CT, TENIS, role, FDG

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3327 The Observable Method for the Regularization of Shock-Interface Interactions

Authors: Teng Li, Kamran Mohseni

Abstract:

This paper presents an inviscid regularization technique that is capable of regularizing the shocks and sharp interfaces simultaneously in the shock-interface interaction simulations. The direct numerical simulation of flows involving shocks has been investigated for many years and a lot of numerical methods were developed to capture the shocks. However, most of these methods rely on the numerical dissipation to regularize the shocks. Moreover, in high Reynolds number flows, the nonlinear terms in hyperbolic Partial Differential Equations (PDE) dominates, constantly generating small scale features. This makes direct numerical simulation of shocks even harder. The same difficulty happens in two-phase flow with sharp interfaces where the nonlinear terms in the governing equations keep sharpening the interfaces to discontinuities. The main idea of the proposed technique is to average out the small scales that is below the resolution (observable scale) of the computational grid by filtering the convective velocity in the nonlinear terms in the governing PDE. This technique is named “observable method” and it results in a set of hyperbolic equations called observable equations, namely, observable Navier-Stokes or Euler equations. The observable method has been applied to the flow simulations involving shocks, turbulence, and two-phase flows, and the results are promising. In the current paper, the observable method is examined on the performance of regularizing shocks and interfaces at the same time in shock-interface interaction problems. Bubble-shock interactions and Richtmyer-Meshkov instability are particularly chosen to be studied. Observable Euler equations will be numerically solved with pseudo-spectral discretization in space and third order Total Variation Diminishing (TVD) Runge Kutta method in time. Results are presented and compared with existing publications. The interface acceleration and deformation and shock reflection are particularly examined.

Keywords: compressible flow simulation, inviscid regularization, Richtmyer-Meshkov instability, shock-bubble interactions.

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3326 The Benefits of Using Transformative Inclusion Practices and Action Research in Teaching Development and Active Participation of Roma Students in the Kindergarten

Authors: Beazidou Eleftheria

Abstract:

Roma children face discrimination in schools where they are the minority. On the other hand, teachers do not identify the specific needs of Roma students for educational and social inclusion and generally use a very restricted repertoire of insufficient strategies for helping them. Modern classrooms can and should look different. Therefore, engaging in transformational learning with young children is a deliberate choice. Transformation implies a different way of thinking and acting. This requires new knowledge that incorporates multiple perspectives and actions in order to generate experiences for further learning. In this way, we build knowledge based on empirical examples, and we share what works efficiently. The present research aims at assisting the participating teachers to improve their teaching inclusive practice, thus ultimately benefiting their students. To increase the impact of transformative efforts with a ‘new’ teaching approach, we implemented a classroom-based action research program for over six months in five kindergarten classrooms with Roma and non-Roma students. More specifically, we explore a) information about participants’ experience of the program and b) if the program is successful in helping participants to change their teaching practice. Action research is, by definition, a form of inquiry that is intended to have both action and research outcomes. The action research process that we followed included five phases: 1. Defining the problem: As teachers said, the Roma students are often the most excluded group in schools (Low social interaction and participation in classroom activities) 2. Developing a plan to address the problem: We decided to address the problem by improving/transforming the inclusive practices that teachers implemented in their classrooms. 3. Acting: implementing the plan: We incorporated new activities for all students with the goals: a) All students being passionate about their learning, b) Teachers must investigate issues in the educational context that are personal and meaningful to children's growth, c) Establishment of a new module for values and skills for all students, d) Raising awareness in culture of Roma, e) Teaching students to reflect. 4. Observing: We explore the potential for transformation in the action research program that involves observations of students’ participation in classroom activities and peer interaction. – thus, generated evidence from data. 5. Reflecting and acting: After analyzing and evaluating the outcomes from data and considering the obstacles during the program’s implementation, we established new goals for the next steps of the program. These are centered in: a) the literacy skills of Roma students and b) the transformation of teacher’s perceptions and believes, which have a powerful impact on their willingness to adopt new teaching strategies. The final evaluation of the program showed a significant achievement of the transformative goals, which were related to the active participation of the Roma students in classroom activities and peer interaction, while the activities which were related to literacy skills did not have the expected results. In conclusion, children were equipped with relevant knowledge and skills to raise their potential and contribute to wider societal development as well as teachers improved their teaching inclusive practice.

Keywords: action research, inclusive practices, kindergarten, transformation

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3325 Animal-Assisted Therapy: A Perspective From Singapore

Authors: Julia Wong, Hua Beng Lim, Petrina Goh, Johanna Foo, Caleb Ng, Nurul ‘Aqilah Bte Mohd Taufek

Abstract:

Animal-assisted therapy (AAT) utilizes human-animal interaction to achieve specific therapeutic goals, and its efficacy has been demonstrated across various settings overseas. The use of AAT in Singapore, however, is still limited. Ang Mo Kio-Thye Hua Kwan (AMKH) is one of the first community hospitals in Singapore to use AAT to complement its occupational therapy services with elderly patients. This study explored the perspectives of AMKH’s occupational therapists (OTs) in relation to AAT to understand barriers and enablers in implementing and practising AAT. We also examined how OTs at-large across practice settings perceive AAT. A mixed method design was used. 64 OTs at-large participated in on online survey, and 7 AMKH OTs were interviewed individually via Zoom. Survey results were analysed with descriptive and Mann-Whitney U tests. Interviews were thematically analysed. AMKH OTs perceived various benefits of AAT articulated in overseas studies in domains such as motivation and participation, emotional, social interaction, sensory tactile stimulation, and cognition. Interestingly, this perception was also supported by 67% of OTs who had responded to the survey, even though most of the OTs who had participated in the survey had no experience in AAT. Despite the perceived benefits of AAT, both OTs from AMKH and those at-large articulated concerns on risks pertaining to AAT (e.g., allergies, unexpected animal behaviour, infections, etc). However, AMKH OTs shared several ways to mitigate these risks, demonstrating their ability to develop a safe program. For e.g., volunteers and their dogs must meet specific recruitment criteria, stringent protocols are used to screen and match dogs with patients, and there are strict exclusion criteria for patients participating in AAT. AMKH OTs’ experience suggests that additional skills and knowledge are required to implement AAT, therefore, healthcare institutions should first consider improving their staff training and risk mitigation knowledge before implementing AAT. They can also refer to AMKH’s AAT protocols and those found in overseas studies, but institutions must adapt the protocols to fit their institutional settings and patients’ profiles.

Keywords: animal-assisted therapy, dog-assisted therapy, occupational therapy, complementary therapy

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3324 Investigating the Post-Liver Transplant Complications and Their Management in Children Referred to the Children’s Medical Center

Authors: Hosein Alimadadi, Fatemeh Farahmand, Ali Jafarian, Nasir Fakhar, Mohammad Hassan Sohouli, Neda Raeesi

Abstract:

Backgroundsː Regarding the important role of liver transplantation as the only treatment in many cases of end-stage liver disease in children, the aim of this study is to investigate the complications of liver transplantation and their management in children referred to the Children's Medical Center. Methods: This study is a cross-sectional study on pediatric patients who have undergone liver transplants in the years 2016 to 2021. The indication for liver transplantation in this population was confirmed by a pediatric gastroenterologist, and a liver transplant was performed by a transplant surgeon. Finally, information about the patient before and after the transplantation was collected and recorded. Results: A total of 53 patients participated in this study, including 25 (47.2%) boys and 28 (52.8%) girls. The most common causes of liver transplantation were cholestatic and metabolic diseases. The most common early complication of liver transplantation in children was acute cellular rejection (ACR) and anastomotic biliary stricture. The most common late complication in these patients was an infection which was observed in 56.6% of patients. Among the drug side effects, neurotoxicity (convulsions) was seen more in patients, and 15.1% of the transplanted patients died. Conclusion: In this study, the most common early complication of liver transplantation in children was ACR and biliary stricture, and the most common late complication was infection. Neurotoxicity (convulsions) was the most common side effect of drugs.

Keywords: liver transplantation, complication, infection, survival rate

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3323 Characterization of Transcription Factors Involved in Early Defense Response during Interaction of Oil Palm Elaeis guineensis Jacq. with Ganoderma boninense

Authors: Sakeh N. Mohd, Bahari M. N. Abdul, Abdullah S. N. Akmar

Abstract:

Oil palm production generates high export earnings to many countries especially in Southeast Asian region. Infection by necrotrophic fungus, Ganoderma boninense on oil palm results in basal stem rot which compromises oil palm production leading to significant economic loss. There are no reliable disease treatments nor promising resistant oil palm variety has been cultivated to eradicate the disease up to date. Thus, understanding molecular mechanisms underlying early interactions of oil palm with Ganoderma boninense may be vital to promote preventive or control measure of the disease. In the present study, four months old oil palm seedlings were infected via artificial inoculation of Ganoderma boninense on rubber wood blocks. Roots of six biological replicates of treated and untreated oil palm seedlings were harvested at 0, 3, 7 and 11 days post inoculation. Next-generation sequencing was performed to generate high-throughput RNA-Seq data and identify differentially expressed genes (DEGs) during early oil palm-Ganoderma boninense interaction. Based on de novo transcriptome assembly, a total of 427,122,605 paired-end clean reads were assembled into 30,654 unigenes. DEGs analysis revealed upregulation of 173 transcription factors on Ganoderma boninense-treated oil palm seedlings. Sixty-one transcription factors were categorized as DEGs according to stringent cut-off values of genes with log2 ratio [Number of treated oil palm seedlings/ Number of untreated oil palm seedlings] ≥ |1.0| (corresponding to 2-fold or more upregulation) and P-value ≤ 0.01. Transcription factors in response to biotic stress will be screened out from abiotic stress using reverse transcriptase polymerase chain reaction. Transcription factors unique to biotic stress will be verified using real-time polymerase chain reaction. The findings will help researchers to pinpoint defense response mechanism specific against Ganoderma boninense.

Keywords: Ganoderma boninense, necrotrophic, next-generation sequencing, transcription factors

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3322 Determination of Biomolecular Interactions Using Microscale Thermophoresis

Authors: Lynn Lehmann, Dinorah Leyva, Ana Lazic, Stefan Duhr, Philipp Baaske

Abstract:

Characterization of biomolecular interactions, such as protein-protein, protein-nucleic acid or protein-small molecule, provides critical insights into cellular processes and is essential for the development of drug diagnostics and therapeutics. Here we present a novel, label-free, and tether-free technology to analyze picomolar to millimolar affinities of biomolecular interactions by Microscale Thermophoresis (MST). The entropy of the hydration shell surrounding molecules determines thermophoretic movement. MST exploits this principle by measuring interactions using optically generated temperature gradients. MST detects changes in the size, charge and hydration shell of molecules and measures biomolecule interactions under close-to-native conditions: immobilization-free and in bioliquids of choice, including cell lysates and blood serum. Thus, MST measures interactions under close-to-native conditions, and without laborious sample purification. We demonstrate how MST determines the picomolar affinities of antibody::antigen interactions, and protein::protein interactions measured from directly from cell lysates. MST assays are highly adaptable to fit to the diverse requirements of different and complex biomolecules. NanoTemper´s unique technology is ideal for studies requiring flexibility and sensitivity at the experimental scale, making MST suitable for basic research investigations and pharmaceutical applications.

Keywords: biochemistry, biophysics, molecular interactions, quantitative techniques

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3321 Functional Gene Expression in Human Cells Using Linear Vectors Derived from Bacteriophage N15 Processing

Authors: Kumaran Narayanan, Pei-Sheng Liew

Abstract:

This paper adapts the bacteriophage N15 protelomerase enzyme to assemble linear chromosomes as vectors for gene expression in human cells. Phage N15 has the unique ability to replicate as a linear plasmid with telomeres in E. coli during its prophage stage of life-cycle. The virus-encoded protelomerase enzyme cuts its circular genome and caps its ends to form hairpin telomeres, resulting in a linear human-chromosome-like structure in E. coli. In mammalian cells, however, no enzyme with TelN-like activities has been found. In this work, we show for the first-time transfer of the protelomerase from phage into human and mouse cells and demonstrate recapitulation of its activity in these hosts. The function of this enzyme is assayed by demonstrating cleavage of its target DNA, followed by detecting telomere formation based on its resistance to recBCD enzyme digestion. We show protelomerase expression persists for at least 60 days, which indicates limited silencing of its expression. Next, we show that an intact human β-globin gene delivered on this linear chromosome accurately retains its expression in the human cellular environment for at least 60 hours, demonstrating its stability and potential as a vector. These results demonstrate that the N15 protelomerse is able to function in mammalian cells to cut and heal DNA to create telomeres, which provides a new tool for creating novel structures by DNA resolution in these hosts.

Keywords: chromosome, beta-globin, DNA, gene expression, linear vector

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