Search results for: Vero cell

Authors: Manuel I. Capel, Antonio Tomeu, Alberto Salguero

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Tumor growth from a transformed cancer-cell up to a clinically apparent mass spans through a range of spatial and temporal magnitudes. Through computer simulations, Cellular Automata (CA) can accurately describe the complexity of the development of tumors. Tumor development prognosis can now be made -without making patients undergo through annoying medical examinations or painful invasive procedures- if we develop appropriate CA-based software tools. In silico testing mainly refers to Computational Biology research studies of application to clinical actions in Medicine. To establish sound computer-based models of cellular behavior, certainly reduces costs and saves precious time with respect to carrying out experiments in vitro at labs or in vivo with living cells and organisms. These aim to produce scientifically relevant results compared to traditional in vitro testing, which is slow, expensive, and does not generally have acceptable reproducibility under the same conditions. For speeding up computer simulations of cellular models, specific literature shows recent proposals based on the CA approach that include advanced techniques, such the clever use of supporting efficient data structures when modeling with deterministic stochastic cellular automata. Multiparadigm and multiscale simulation of tumor dynamics is just beginning to be developed by the concerned research community. The use of stochastic cellular automata (SCA), whose parallel programming implementations are open to yield a high computational performance, are of much interest to be explored up to their computational limits. There have been some approaches based on optimizations to advance in multiparadigm models of tumor growth, which mainly pursuit to improve performance of these models through efficient memory accesses guarantee, or considering the dynamic evolution of the memory space (grids, trees,…) that holds crucial data in simulations. In our opinion, the different optimizations mentioned above are not decisive enough to achieve the high performance computing power that cell-behavior simulation programs actually need. The possibility of using multicore and GPU parallelism as a promising multiplatform and framework to develop new programming techniques to speed-up the computation time of simulations is just starting to be explored in the few last years. This paper presents a model that incorporates parallel processing, identifying the synchronization necessary for speeding up tumor growth simulations implemented in Java and C++ programming environments. The speed up improvement that specific parallel syntactic constructs, such as executors (thread pools) in Java, are studied. The new tumor growth parallel model is proved using implementations with Java and C++ languages on two different platforms: chipset Intel core i-X and a HPC cluster of processors at our university. The parallelization of Polesczuk and Enderling model (normally used by researchers in mathematical oncology) proposed here is analyzed with respect to performance gain. We intend to apply the model and overall parallelization technique presented here to solid tumors of specific affiliation such as prostate, breast, or colon. Our final objective is to set up a multiparadigm model capable of modelling angiogenesis, or the growth inhibition induced by chemotaxis, as well as the effect of therapies based on the presence of cytotoxic/cytostatic drugs.

Keywords: cellular automaton, tumor growth model, simulation, multicore and manycore programming, parallel programming, high performance computing, speed up

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Authors: Wiam El Kheir, Anais Dumais, Maude Beaudoin, Bernard Marcos, Nick Virgilio, Benoit Paquette, Nathalie Faucheux, Marc-Antoine Lauzon

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The high infiltrative pattern of glioblastoma multiforme cells (GBM) is the main cause responsible for the actual standard treatments failure. The tumor high heterogeneity, the interstitial fluid flow (IFF) and chemokines guides GBM cells migration in the brain parenchyma resulting in tumor recurrence. Drug delivery systems emerged as an alternative approach to develop effective treatments for the disease. Some recent studies have proposed to harness the effect CXC-lchemokine-ligand-12 to direct and control the cancer cell migration through delivery system. However, the dynamics of the brain environment on the delivery system remains poorly understood. Nanoparticles (NPs) and hydrogels are known as good carriers for the encapsulation of different agents and control their release. We studied the release of CXCL12 (free or loaded into NPs) from an alginate-based hydrogel under static and indirect perfusion (IP) conditions. Under static conditions, the main phenomena driving CXCL12 release from the hydrogel was diffusion with the presence of strong interactions between the positively charged CXCL12 and the negatively charge alginate. CXCL12 release profiles were independent from the initial mass loadings. Afterwards, we demonstrated that the release could tuned by loading CXCL12 into Alginate/Chitosan-Nanoparticles (Alg/Chit-NPs) and embedded them into alginate-hydrogel. The initial burst release was substantially attenuated and the overall cumulative release percentages of 21%, 16% and 7% were observed for initial mass loadings of 0.07, 0.13 and 0.26 µg, respectively, suggesting stronger electrostatic interactions. Results were mathematically modeled based on Fick’s second law of diffusion framework developed previously to estimate the effective diffusion coefficient (Deff) and the mass transfer coefficient. Embedding the CXCL12 into NPs decreased the Deff an order of magnitude, which was coherent with experimental data. Thereafter, we developed an in-vitro 3D model that takes into consideration the convective contribution of the brain IFF to study CXCL12 release in an in-vitro microenvironment that mimics as faithfully as possible the human brain. From is unique design, the model also allowed us to understand the effect of IP on CXCL12 release in respect to time and space. Four flow rates (0.5, 3, 6.5 and 10 µL/min) which may increase CXCL12 release in-vivo depending on the tumor location were assessed. Under IP, cumulative percentages varying between 4.5-7.3%, 23-58.5%, 77.8-92.5% and 89.2-95.9% were released for the three initial mass loadings of 0.08, 0.16 and 0.33 µg, respectively. As the flow rate increase, IP culture conditions resulted in a higher release of CXCL12 compared to static conditions as the convection contribution became the main driving mass transport phenomena. Further, depending on the flow rate, IP had a direct impact on CXCL12 distribution within the simulated brain tissue, which illustrates the importance of developing such 3D in-vitro models to assess the efficiency of a delivery system targeting the brain. In future work, using this very model, we aim to understand the impact of the different phenomenon occurring on GBM cell behaviors in response to the resulting chemokine gradient subjected to various flow while allowing them to express their invasive characteristics in an in-vitro microenvironment that mimics the in-vivo brain parenchyma.

Keywords: 3D culture system, chemokines gradient, glioblastoma multiforme, kinetic release, mathematical modeling

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Authors: V. Naga Sravan Kumar Varma, H. G. Shivakumar

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The aim of the study was to develop dual sensitive poly (N-isopropylacrylamide-co-acrylic acid) (PNA) nanogels(NGs) and studying its applications for Anti-Cancer therapy. NGs were fabricated by free radical polymerization using different amount of N-isopropylacrylamide and acrylic acid. A study for polymer composition over the effect on LCST in different pH was evaluated by measuring the absorbance at 500nm using UV spectrophotometer. Further selected NG’s were evaluated for change in hydrodynamic diameters in response to pH and temperature. NGs which could sharply respond to low pH value of cancer cells at body temperature were loaded with Fluorouracil (5-FU) using equilibrium swelling method and studied for drug release behaviour in different pH. A significant influence of NGs polymer composition over pH dependent LCST was observed. NGs which were spherical with an average particle size of 268nm at room temperature, shrinked forming an irregular shape when heated above to their respective LCST. 5FU loaded NGs did not intervene any difference in pH depended LCST behaviour of NGs. The in vitro drug release of NGs exhibited a pH and thermo-dependent control release. The cytoxicity study of blank carrier to MCF7 cell line showed no cytotoxicity. The results indicated that PNA NGs could be used as a potential drug carrier for anti-cancer therapy.

Keywords: pH and thermo-sensitive, nanogels, P(NIPAM-co-AAc), anti-cancer, 5-FU

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Authors: Kambiz Hassanzadeh, Seyedeh Shohreh Ebrahimi, Shahrbanoo Oryan, Arman Rahimmi, Esmael Izadpanah

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Parkinson’s disease is one of the most prevalent neurodegenerative diseases which occurs in elderly. There are convincing evidences that oxidative stress has an important role in both the initiation and progression of Parkinson’s disease. Thymoquinone (TQ) is shown to have antioxidant and anti-inflammatory properties in invitro and invivo studies. It is well documented that TQ acts as a free radical scavenger and prevents the cell damage. Therefore this study aimed to evaluate the effect of TQ on motor and non-motor symptoms in animal model of Parkinson’s disease. Male Wistar rats (10-12 months) received rotenone (1mg/kg/day, sc) to induce Parkinson’s disease model. Pretreatment with TQ (7.5 and 15 mg/kg/day, po) was administered one hour before the rotenone injection. Three motor tests (rotarod, rearing and bar tests) and two non-motor tests (forced swimming and elevated plus maze) were performed for behavioral assessment. Our results indicated that TQ significantly ameliorated the rotenone-induced motor dysfunction in rotarod and rearing tests also it could prevent the non-motor dysfunctions in forced swimming and elevated plus maze tests. In conclusion we found that TQ delayed the Parkinson's disease induction by rotenone and this effect might be related to its proved antioxidant effect.

Keywords: Parkinson's disease, thymoquinone, motor and non-motor symptoms, neurodegenerative disease

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Authors: Jaeseung Lee, Muhammad Faizan Chinannai, Mohamed Hassan Gundu, Hyunchul Ju

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In this paper, we numerically investigated the effect of metal foams on a real scale 242.57cm2 (19.1 cm × 12.7 cm) polymer electrolyte membrane fuel cell (PEFCs) using a three-dimensional two-phase PEFC model to substantiate design approach for PEFCs using metal foam as the flow distributor. The simulations were conducted under the practical low humidity hydrogen, and air gases conditions in order to observe the detailed operation result in the PEFCs using the serpentine flow channel in the anode and metal foam design in the cathode. The three-dimensional contours of flow distribution in the channel, current density distribution in the membrane and hydrogen and oxygen concentration distribution are provided. The simulation results revealed that the use of highly porous and permeable metal foam can be beneficial to achieve a more uniform current density distribution and better hydration in the membrane under low inlet humidity conditions. This study offers basic directions to design channel for optimal water management of PEFCs.

Keywords: polymer electrolyte fuel cells, metal foam, real-scale, numerical model

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Authors: Dong Xie, Yuling Xu, Leah Howard

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Secondary caries is found to be one of the main reasons to the restoration failure of dental restoratives. To prevent secondary caries formation, dental restoratives ought to be made antibacterial. In this study, a natural fruit component furaneol was tethered onto polyacid, the formed polyacid was used to formulate the light-curable glass-ionomer cements, and then the effect of this new antibacterial compound on compressive strength (CS) and antibacterial activity of the formed cement was evaluated. Fuji II LC glass powders were used as fillers. Compressive strength (CS) and S. mutans viability were used to evaluate the mechanical strength and antibacterial activity of the formed cement. The experimental cement showed a significant antibacterial activity, accompanying with an initial CS reduction. Increasing the compound loading significantly decreased the S. mutans viability from 5 to 81% and also reduced the initial CS of the formed cements from 4 to 58%. The cement loading with 7% antibacterial polymer showed 168 MPa, 7.8 GPa, 243 MPa, 46 MPa, and 57 MPa in yield strength, modulus, CS, diametral tensile strength and flexural strength, respectively, as compared to 141, 6.9, 236, 42 and 53 for Fuji II LC. The cement also showed an antibacterial function to other bacteria. No human saliva effect was noticed. It is concluded that the experimental cement may potentially be developed to a permanent antibacterial cement.

Keywords: antibacterial, dental materials, strength, cell viability

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Authors: Huayang Yu, Nicola Ingram, David C. Green, Paul D. Thornton

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The dual stimuli-controlled release of doxorubicin from gel-embedded nanoparticles is reported. Non-cytotoxic polymer nanoparticles are formed from poly(ethylene glycol)-b-poly(benzyl glutamate) that, uniquely, contain a central ester link. This connection renders the nanoparticles pH-responsive, enabling extensive doxorubicin release in acidic solutions (pH 6.5), but not in solutions of physiological pH (pH 7.4). Doxorubicin loaded nanoparticles were found to be stable for at least 31 days and lethal against the three breast cancer cell lines tested. Furthermore, doxorubicin-loaded nanoparticles could be incorporated within a thermoresponsive poly(2-hydroxypropyl methacrylate) gel depot, which forms immediately upon injection of poly(2-hydroxypropyl methacrylate) into aqueous solution. The combination of the poly(2-hydroxypropyl methacrylate) gel and poly(ethylene glycol)-b-poly(benzyl glutamate) nanoparticles yields an injectable doxorubicin delivery system that facilities near-complete drug release when maintained at elevated temperatures (37 °C) in acidic solution (pH 6.5). In contrast, negligible payload release occurs when the material is stored at room temperature in a non-acidic solution (pH 7.4). The system has great potential as a vehicle for the prolonged, site-specific release of chemotherapeutics.

Keywords: biodegradable, nanoparticle, polymer, thermoresponsive

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Authors: Jaden Lu, Olivia Lu

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Despite the remarkable advancement of solar cell technology, the challenge of optimizing total solar energy harvest efficiency persists, primarily due to significant heat loss. This excess heat not only diminishes solar panel output efficiency but also curtails its operational lifespan. A promising approach to address this issue is the conversion of surplus heat into electricity. In recent years, there is growing interest in the use of thermoelectric generators (TEG) as a potential solution. The integration of efficient TEG devices holds the promise of augmenting overall energy harvest efficiency while prolonging the longevity of solar panels. While certain research groups have proposed the integration of solar cells and TEG devices, a substantial gap between conceptualization and practical implementation remains, largely attributed to low thermal energy conversion efficiency of TEG devices. To bridge this gap and meet the requisites of practical application, a feasible strategy involves the incorporation of a substantial number of p-n junctions within a confined unit volume. However, the manufacturing of high-density TEG p-n junctions presents a formidable challenge. The prevalent solution often leads to large device sizes to accommodate enough p-n junctions, consequently complicating integration with solar cells. Recently, the adoption of 3D printing technology has emerged as a promising solution to address this challenge by fabricating high-density p-n arrays. Despite this, further developmental efforts are necessary. Presently, the primary focus is on the 3D printing of vertically layered TEG devices, wherein p-n junction density remains constrained by spatial limitations and the constraints of 3D printing techniques. This study proposes a novel device configuration featuring horizontally arrayed p-n junctions of Bi2Te3. The structural design of the device is subjected to simulation through the Finite Element Method (FEM) within COMSOL Multiphysics software. Various device configurations are simulated to identify optimal device structure. Based on the simulation results, a new TEG device is fabricated utilizing 3D Selective laser melting (SLM) printing technology. Fusion 360 facilitates the translation of the COMSOL device structure into a 3D print file. The horizontal design offers a unique advantage, enabling the fabrication of densely packed, three-dimensional p-n junction arrays. The fabrication process entails printing a singular row of horizontal p-n junctions using the 3D SLM printing technique in a single layer. Subsequently, successive rows of p-n junction arrays are printed within the same layer, interconnected by thermally conductive copper. This sequence is replicated across multiple layers, separated by thermal insulating glass. This integration created in a highly compact three-dimensional TEG device with high density p-n junctions. The fabricated TEG device is then attached to the bottom of the solar cell using thermal glue. The whole device is characterized, with output data closely matching with COMSOL simulation results. Future research endeavors will encompass the refinement of thermoelectric materials. This includes the advancement of high-resolution 3D printing techniques tailored to diverse thermoelectric materials, along with the optimization of material microstructures such as porosity and doping. The objective is to achieve an optimal and highly integrated PV-TEG device that can substantially increase the solar energy harvest efficiency.

Keywords: thermoelectric, finite element method, 3d print, energy conversion

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Authors: Katarzyna Lubecka, Kirsty Flower, Megan Beetch, Lucinda Kurzava, Hannah Buvala, Samer Gawrieh, Suthat Liangpunsakul, Tracy Gonzalez, George McCabe, Naga Chalasani, James M. Flanagan, Barbara Stefanska

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Hepatocellular carcinoma (HCC), the most prevalent type of primary liver cancer, is the second leading cause of cancer death worldwide. Late onset of clinical symptoms in HCC results in late diagnosis and poor disease outcome. Approximately 85% of individuals with HCC have underlying liver cirrhosis. However, not all cirrhotic patients develop cancer. Reliable early detection biomarkers that can distinguish cirrhotic patients who will develop cancer from those who will not are urgently needed and could increase the cure rate from 5% to 80%. We used Illumina-450K microarray to test whether blood DNA, an easily accessible source of DNA, bear site-specific changes in DNA methylation in response to HCC before diagnosis with conventional tools (pre-diagnostic). Top 11 differentially methylated sites were selected for validation by pyrosequencing. The diagnostic potential of the 11 pyrosequenced probes was tested in blood samples from a prospective cohort of cirrhotic patients. We identified 971 differentially methylated CpG sites in pre-diagnostic HCC cases as compared with healthy controls (P < 0.05, paired Wilcoxon test, ICC ≥ 0.5). Nearly 76% of differentially methylated CpG sites showed lower levels of methylation in cases vs. controls (P = 2.973E-11, Wilcoxon test). Classification of the CpG sites according to their location relative to CpG islands and transcription start site revealed that those hypomethylated loci are located in regulatory regions important for gene transcription such as CpG island shores, promoters, and 5’UTR at higher frequency than hypermethylated sites. Among 735 CpG sites hypomethylated in cases vs. controls, 482 sites were assigned to gene coding regions whereas 236 hypermethylated sites corresponded to 160 genes. Bioinformatics analysis using GO, KEGG and DAVID knowledgebase indicate that differentially methylated CpG sites are located in genes associated with functions that are essential for gene transcription, cell adhesion, cell migration, and regulation of signal transduction pathways. Taking into account the magnitude of the difference, statistical significance, location, and consistency across the majority of matched pairs case-control, we selected 11 CpG loci corresponding to 10 genes for further validation by pyrosequencing. We established that methylation of CpG sites within 5 out of those 10 genes distinguish cirrhotic patients who subsequently developed HCC from those who stayed cancer free (cirrhotic controls), demonstrating potential as biomarkers of early detection in populations at risk. The best predictive value was detected for CpGs located within BARD1 (AUC=0.70, asymptotic significance ˂0.01). Using an additive logistic regression model, we further showed that 9 CpG loci within those 5 genes, that were covered in pyrosequenced probes, constitute a panel with high diagnostic accuracy (AUC=0.887; 95% CI:0.80-0.98). The panel was able to distinguish pre-diagnostic cases from cirrhotic controls free of cancer with 88% sensitivity at 70% specificity. Using blood as a minimally invasive material and pyrosequencing as a straightforward quantitative method, the established biomarker panel has high potential to be developed into a routine clinical test after validation in larger cohorts. This study was supported by Showalter Trust, American Cancer Society (IRG#14-190-56), and Purdue Center for Cancer Research (P30 CA023168) granted to BS.

Keywords: biomarker, DNA methylation, early detection, hepatocellular carcinoma

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Authors: Yomna Ibrahim El-Gazzar, Hussien Ibrahim El-Subbagh, Hanan Hanaa Georgey, Ghada S. Hassan Hassan

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Dihydrofolate reductase (DHFR) is an enzyme that has pivotal importance in biochemistry and medicinal chemistry. It catalyzes the reduction of dihydrofolate to tetrahydrofolate and intimately couples with thymidylate synthase. Thymidylate synthase is a crucial enzyme that catalyzes the reductive methylation of (dUMP) to (dTMP) utilizing N5, N10-methylenetetrahydrofolate as a cofactor. A new series of 2-substituted thio-quinazolin-4-one analogs was designed that possessed electron withdrawing or donating functional groups (Cl or OCH3) at position 6- or 7-, 4-methoxyphenyl function at position 3-.The thiol function is used to connect to either 1,2,4-triazole, or 1,3,4-thiadiazole via a methylene bridge. Most of the functional groups designed to be accommodated on the quinazoline ring such as thioether, alkyl to increase lipid solubility of polar compounds, a character very much needed in the nonclassical DHFR inhibitors. The target compounds were verified with spectral data and elemental analysis. DHFR inhibitions, as well as antitumor activity, were applied on three cell lines (MCF-7, CACO-2, HEPG-2).

Keywords: nonclassical antifolates, DHFR Inhibitors, antitumor activity, quinazoline ring

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Authors: Sagarjyoti Roy

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Dengue is an arboviral infection, belonging to family flaviviridae, comprising of four serotypes; DENV1, DENV2, DENV3 and DENV4. All four serotypes are capable of causing full-spectrum of clinical features, ranging from self-limiting fever to severe dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). Complications may affect any organ system, including those involving gastrointestinal system and serositis. We report a case, of a 28 years, non-alcoholic male, presenting with a 7 day history of fever and malaise followed by abdominal pain and distension, from 4th day of fever. He was admitted in medicine department of RG KAR medical college hospital. Dengue fever was confirmed by NS1 and dengue IgM positivity. Platelet count was 30,000/cc (1.5- 4 lac/cc) and haematocrit was 52% (38- 50% for men). Clinicoradiological findings revealed bilateral pleural effusion, ascites and splenomegaly. Ascitic fluid was hemorrhagic in nature, with a high protein and RBC content. Liver function tests revealed mild transaminitis with normal coagulation profile. Patient was managed conservatively. A diagnosis of dengue fever complicated by serositis and spontaneous haemoperitoneum was made. The symptoms subsided after a hospital stay of 10 days. The case highlights haemorrhage into peritoneal cavity as a possible complication of dengue fever. Although a definite explanation requires more detailed studies, platelet or endothelial cell dysfunction might be contributory.

Keywords: ascites, dengue, haemoperitoneum, serositis

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Authors: Sadaf Ilyas

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Recombinant cytokines have been employed successfully as potential therapeutic agent. Some cytokine therapies are already used as a part of clinical practice, ranging from early exploratory trials to well established therapies that have already received approval. Interleukin 15 is a pleiotropic cytokine having multiple roles in peripheral innate and adaptive immune cell function. It regulates the activation, proliferation and maturation of NK cells, T-cells, monocytes/macrophages and granulocytes, and the interactions between them thus acting as a bridge between innate and adaptive immune responses. Unraveling the biology of IL-15 has revealed some interesting surprises that may point toward some of the first therapeutic applications for this cytokine. In this study, the human interleukin 15 gene was isolated, amplified and ligated to a TA vector which was then transfected to a bacterial host, E. coli Top10F’. The sequence of cloned gene was confirmed and it showed 100% homology with the reported sequence. The confirmed gene was then subcloned in pET Expression system to study the IPTG induced expression of IL-15 gene. Positive expression was obtained for number of clones that showed 15 kd band of IL-15 in SDS-PAGE analysis, indicating the successful strain development that can be studied further to assess the potential therapeutic intervention of this cytokine in relevance to human diseases.

Keywords: Interleukin 15, pET expression system, immune therapy, protein purification

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Authors: Subrata Kar, Banani Kundu, Papiya Nandy, Ruma Basu, Sukhen Das

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Natural montmorilollite clay is a common ingredient in pharmaceutical products, both as excipients and active support; hence considered as suitable candidate for Drug Delivery System. In this work, cationic detergent CTAB is used to increase the interlayer spacing of Na+-Montmoriollite clay to intercalate curcumin and methotrexate. Methotrexate is a folic acid antagonist, anti-proliferative and immunosuppressive agent; while curcumin is a bioactive constituent of rhizomes of Curcuma longa, possessing remarkable chemo-preventive and anti-inflammatory properties. The resultant inorganic-organic hybrids are characterized by X-ray diffraction (XRD), Infrared spectroscopy (FTIR) and Thermo Gravimetric Analysis (TGA) to confirm successful intercalation of curcumin and Methotrexate within clay layers. Pharmaceutical investigation of the hybrids is explored by studying the drug loading (%), encapsulation efficiency and release kinetics. Finally in-vitro studies are performed using cancer cells to find the effect of released curcumin to improve the sensitivity of clay bound methotrexate to ameliorate cell death compared to their effectiveness when used without the inorganic aluminosilicate vehicle.

Keywords: montmorillonite, methotrexate, curcumin, loading efficiency, release kinetics, anticancer activity

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Authors: Santosh Kumar Biswal, Ramakar Jha

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Soil erosion is a slow and continuous process and one of the prominent problems across the world leading to many serious problems like loss of soil fertility, loss of soil structure, poor internal drainage, sedimentation deposits etc. In this paper remote sensing and GIS based methods have been applied for the determination of soil erosion and sediment yield. Tel River basin which is the second largest tributary of the river Mahanadi laying between latitude 19° 15' 32.4"N and, 20° 45' 0"N and longitude 82° 3' 36"E and 84° 18' 18"E chosen for the present study. The catchment was discretized into approximately homogeneous sub-areas (grid cells) to overcome the catchment heterogeneity. The gross soil erosion in each cell was computed using Universal Soil Loss Equation (USLE). Various parameters for USLE was determined as a function of land topography, soil texture, land use/land cover, rainfall, erosivity and crop management and practice in the watershed. The concept of transport limited accumulation was formulated and the transport capacity maps were generated. The gross soil erosion was routed to the catchment outlet. This study can help in recognizing critical erosion prone areas of the study basin so that suitable control measures can be implemented.

Keywords: Universal Soil Loss Equation (USLE), GIS, land use, sediment yield,

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Authors: Kah Yan How, Peh Fern Ong, Keang Peng Song

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Porphyromonas gingivalis is a black-pigmented, anaerobic Gram-negative bacterium that is important in the progression of chronic and severe periodontitis. This organism has an essential requirement for iron, which is usually obtained from hemin, using specific membrane receptors, proteases, and lipoproteins. In this study, we report the characterization of a novel 24 kDa hemin-binding protein, HmuX, in P. gingivalis W50. The hmuX gene is 651 bp long which encodes for a 217 amino acid protein. HmuX was found to be identical at the C-terminus to the previously reported HmuY protein, differing by an additional 74 amino acids at the N-terminus. Recombinant HmuX demonstrated hemin-binding ability by LDS- PAGE and TMBZ staining. Sequence analysis of HmuX revealed a putative lipoprotein attachment site, suggesting its possible role as a lipoprotein. HmuX was also localized to the outer cell surface by transmission electron microscopy. Northern analysis showed hmuX to be transcribed as a single gene and that hmuX mRNA was tightly regulated by the availability of extra-cellular hemin. P. gingivalis isogenic mutant deficient in hmuX gene exhibited significant growth retardation under hemin-limited conditions. Taken together, these results suggest that HmuX is a hemin-binding lipoprotein, important in hemin utilization for the growth of P. gingivalis.

Keywords: Porphyromonas gingivalis, periodontal diseases, HmuX, protein characterization

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Authors: Sudhir Kumar Sharma, Ramesh Jagannathan

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The interaction of organic and inorganic matrices of biological origin resulting in self-assembled structures with unique properties is well established. The development of such self-assembled nanostructures by synthetic and bio-inspired techniques is an established field of active research. Among bio-materials, nacre, a laminar stack of calcium carbonate nanosheets, which are interleaved with organic material, has long been focused research due to its unique mechanical properties. In this paper, we present the development of nacre-like lamellar structures made up of calcium carbonate via a wet chemical route. We used the binding affinity of carboxylate anions and calcium cations using poly (acrylic) acid (PAA) to lead CaCO₃ crystallization. In these experiments, we selected calcium acetate as the precursor molecule along with PAA (Mw ~ 8000 Da). We found that Ca⁺²/COO⁻ ratio provided a tunable control for the morphology and growth of CaCO₃ nanostructures. Drop casting one such formulation on a silicon substrate followed by calcination resulted in co-planner, molecular sheets of CaCO₃, separated by a spacer layer of carbon. The scope of our process could be expanded to produce unit cell thick molecular sheets of other important inorganic materials.

Keywords: self-assembled structures, bio-inspired materials, calcium carbonate, wet chemical route

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Authors: Minzi Mao, Jianjun Ren, Yu Zhao

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Objectives: The absence of a uniform staging scheme for temporal bone carcinoma (TBC) seriously impedes the improvement of its management strategies. Therefore, this research was aimed to investigate the prognostic values of two currently applying staging schemes, namely, the modified Pittsburgh staging system (MPB) and Stell’s T classification (Stell-T) in patients with TBC. Methods: Areal-world single-institution retrospectivereview of patientsdiagnosed with TBC between2008 and 2019 was performed. Baseline characteristics were extracted, and patients were retrospectively staged by both the MPB and Stell-T classifications. Cox regression analyseswereconductedtocomparetheoverall survival (OS). Results: A total of 69 consecutive TBC patients were included in thisstudy. Univariate analysis showed that both Stell-T and T- classifications of the modified Pittsburgh staging system (MPB-T) were significant prognostic factors for all TBC patients as well as temporal bone squamous cell carcinoma (TBSCC, n=50) patients (P < 0.05). However, only Stell-T was confirmed to be an independent prognostic factor in TBSCC patients (P = 0.004). Conclusions: Tumor extensions, quantified by both Stell-T and MPB-T classifications, are significant prognostic factors for TBC patients, especially for TBSCC patients. However, only the Stell-T classification is an independent prognostic factor for TBSCC patients.

Keywords: modified pittsburgh staging system, overall survival, prognostic factor, stell’s T- classification, temporal bone carcinoma

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Authors: Aslihan Cetinbas Genc, Meral Unal

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This study presents the structural and cytochemical alterations of stigma at the stages of pre-anthesis, anthesis and post-anthesis in Vibirnum tinus. Capitate stigma continues with a closed style. The receptive surface of stigma is composed of unicellular papillae which are short and flattened at pre-anthesis stage. The papillae in this stage have dense cytoplasm with small vacuoles and a centrally located nucleus. With the start of anthesis, the stigma widens, papillae lengthen and become cylindrical. At anthesis stage, vacuoles enlarge, and nucleus moves to the base of the cell. At post-anthesis stage, the boundaries of the papillae become less noticeable. As proved by Periodic Acid Schiff procedure, the cytoplasm of papillae is rich in insoluble polysaccharides at all stages of development but it becomes remarkable at post-anthesis, particularly at the sub-papillar area. Although there is no significant difference in the content of protein in all stages of the development, it is more abundant at post-anthesis stage, as in Coomassie Brillant Blue stained sections. The surface of papillae is covered by a cuticle which becomes thicker at post-anthesis, and it gives positive reaction with Sudan Black B and Auramine O. The cuticle is covered by a pellicle stained by Coomassie Brillant Blue, indicating dry type of stigma.

Keywords: develeopmental features, histochemistry, stigma, Vibirnum tinus

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Authors: Vanja Misic, Mohamed El-Mogy, Yousef Haj-Ahmad

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Endonuclease G (EndoG) is a well conserved mitochondrio-nuclear nuclease with dual lethal and vital roles in the cell. The aim of our study was to examine whether EndoG exerts its nuclease activity on exogenous DNA substrates such as plasmid DNA (pDNA), considering their importance in gene therapy applications. The effects of EndoG knockdown on pDNA stability and levels of encoded reporter gene expression were evaluated in the cervical carcinoma HeLa cells. Transfection of pDNA vectors encoding short-hairpin RNAs (shRNAs) reduced levels of EndoG mRNA and nuclease activity in HeLa cells. In physiological circumstances, EndoG knockdown did not have an effect on the stability of pDNA or the levels of encoded transgene expression as measured over a four day time-course. However, when endogenous expression of EndoG was induced by an extrinsic stimulus, targeting of EndoG by shRNA improved the perceived stability and transgene expression of pDNA vectors. Therefore, EndoG is not a mediator of exogenous DNA clearance, but in non-physiological circumstances it may non-specifically cleave intracellular DNA regardless of its origin. These findings make it unlikely that targeting of EndoG is a viable strategy for improving the duration and level of transgene expression from non-viral DNA vectors in gene therapy efforts.

Keywords: EndoG, silencing, exogenous DNA stability, HeLa cells

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Authors: Khrizelle Angelique Sablan, Rizalinda De Leon, Jaeyoung Lee, Joey Ocon

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The impending water shortage drives us to find alternative sources of water. One of the possible solutions is desalination of seawater. There are numerous processes by which it can be done and one if which is capacitive deionization. Capacitive deionization is a relatively new technique for water desalination. It utilizes the electric double layer for ion adsorption. Carbon-based materials are commonly used as electrodes for capacitive deionization. In this study, carbon nanotubes (CNTs) were treated in a mixture of nitric and sulfuric acid. The silver addition was also facilitated to incorporate antimicrobial action. The acid-treated carbon nanotubes (f-CNTs) and silver-decorated f-CNTs (Ag@f-CNTs) were used as electrode materials for seawater deionization and compared with CNT and acid-treated CNT. The synthesized materials were characterized using TEM, EDS, XRD, XPS and BET. The electrochemical performance was evaluated using cyclic voltammetry, and the deionization performance was tested on a single cell with water containing 64mg/L NaCl. The results showed that the synthesized Ag@f-CNT-10 H could have better performance than CNT and a-CNT with a maximum ion removal efficiency of 50.22% and a corresponding adsorption capacity of 3.21 mg/g. It also showed antimicrobial activity against E. coli. However, the said material lacks stability as the efficiency decreases with repeated usage of the electrode.

Keywords: capacitive deionization, carbon nanotubes, desalination, acid functionalization, silver

Procedia PDF Downloads 229

Authors: Cheng Zhang, James Marco, Walid Allafi, Truong Q. Dinh, W. D. Widanage

Abstract:

Equivalent circuit models (ECMs) are widely used in battery management systems in electric vehicles and other battery energy storage systems. The battery dynamics and the model parameters vary under different working conditions, such as different temperature and state of charge (SOC) levels, and therefore online parameter identification can improve the modelling accuracy. This paper presents a way of online ECM parameter identification using a continuous time (CT) estimation method. The CT estimation method has several advantages over discrete time (DT) estimation methods for ECM parameter identification due to the widely separated battery dynamic modes and fast sampling. The presented method can be used for online SOC estimation. Test data are collected using a lithium ion cell, and the experimental results show that the presented CT method achieves better modelling accuracy compared with the conventional DT recursive least square method. The effectiveness of the presented method for online SOC estimation is also verified on test data.

Keywords: electric circuit model, continuous time domain estimation, linear integral filter method, parameter and SOC estimation, recursive least square

Procedia PDF Downloads 381

Authors: Arvind Kumar, Manoj Kumar, Dattatraya H. Nagaraj, Amanpreet Singh, Jayanthi Prattapati

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The purpose of this paper is to study and compare two maximum power point tracking (MPPT) algorithms in a photovoltaic simulation system and also show a simulation study of maximum power point tracking (MPPT) for photovoltaic systems using perturb and observe algorithm and Incremental conductance algorithm. Maximum power point tracking (MPPT) plays an important role in photovoltaic systems because it maximize the power output from a PV system for a given set of conditions, and therefore maximize the array efficiency and minimize the overall system cost. Since the maximum power point (MPP) varies, based on the irradiation and cell temperature, appropriate algorithms must be utilized to track the (MPP) and maintain the operation of the system in it. MATLAB/Simulink is used to establish a model of photovoltaic system with (MPPT) function. This system is developed by combining the models established of solar PV module and DC-DC Boost converter. The system is simulated under different climate conditions. Simulation results show that the photovoltaic simulation system can track the maximum power point accurately.

Keywords: incremental conductance algorithm, perturb and observe algorithm, photovoltaic system, simulation results

Procedia PDF Downloads 554

Authors: M. Sumida, S. Morita

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In this research, the effect of unsteady flows acting on road vehicles was experimentally investigated, using an advanced and recently introduced wind tunnel. The aims of this study were to extract the characteristics of fluid forces acting on road vehicles under unsteady wind conditions and obtain new information on drag forces in a practical on-road test. We applied pulsating wind as a representative example of the atmospheric fluctuations that vehicles encounter on the road. That is, we considered the case where the vehicles are moving at constant speed in the air, with large wind oscillations. The experimental tests were performed on the Ahmed-type test model, which is a simplified vehicle model. This model was chosen because of its simplicity and the data accumulated under steady wind conditions. The experiments were carried out with a time-averaged Reynolds number of Re = 4.16x10⁵ and a pulsation period of T = 1.5 s, with amplitude of η = 0.235. Unsteady fluid forces of drag and lift were obtained utilizing a multi-component load cell. It was observed that the unsteady aerodynamic forces differ significantly from those under steady wind conditions. They exhibit a phase shift and an enhanced response to the wind oscillations. Furthermore, their behavior depends on the slant angle of the rear shape of the model.

Keywords: Ahmed body, automotive aerodynamics, unsteady wind, wind tunnel test

Procedia PDF Downloads 292

Authors: Khairy Mohamed Abdalla Zoheir

Abstract:

One of the most severe complications of Rheumatoid arthritis is delayed recovery. lipoic acid possesses antioxidant, hypoglycemic, and anti-inflammatory activity. In the present study, the effects of lipoic acid were investigated on the key mediators of Rheumatoid arthritis, namely, CD4+CD25+ T cell subsets, GITR expressing cells, CD4+CD25+Foxp3+ regulatory T (Treg) cells, T-helper-17 (Th17) cells and pro-inflammatory cytokines Interleukin-1β (IL-1β), Interleukin-6 (IL-6) and Tumor Necrosis Factor- α (TNF-α)] through flow-cytometry and qPCR analyses. Lipoic acid-treated mice showed a significant decrease in Rheumatoid arthritis, the frequency of GITR-expressing cells, and Th1 cytokines (IL-17A, TNF-αand Interferon- γ (IFN-γ) compared with positive and negative controlled mice. Lipoic acid treatment also downregulated the mRNA expression of the inflammatory mediators compared with the Rheumatoid arthritis mouse model and untreated mice. The number of Tregs was also found to be significantly upregulated in lipoic acid-treated mice. Our results were confirmed by the histopathological examination. This study showed the beneficial role of lipoic acid in promoting a well-balanced tool for the therapy of Rheumatoid arthritis.

Keywords: lipoic acid, inflammatory markers, rheumatoid arthritis, qPCR

Procedia PDF Downloads 98

Authors: Eunhwa Jun

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This case report describes the misdiagnosis of a patient who presented with right arm pain as cervical disc herniation. The patient had several underlying conditions, including hypertension, diabetes mellitus, liver cirrhosis, a history of lung cancer with left lower lobe lobectomy, and adjuvant chemoradiotherapy. An external cervical spine MRI revealed central protruding discs at the C4-5-6-7 levels. Despite treatment with medication and epidural blocks, the patient's pain persisted. A C-RACZ procedure was planned, but the patient's pain had worsened before admission. Using ultrasound, a brachial plexus block was attempted, but the brachial plexus eluded clear visualization, hinting at underlying neurological complexities. Chest CT revealed a new, large soft tissue mass in the right supraclavicular region with adjacent right axillary lymphadenopathy, leading to the diagnosis of metastatic squamous cell carcinoma. Palliative radiation therapy and chemotherapy were initiated as part of the treatment plan, and the patient's pain score decreased to 3 out of 10 on the Numeric Rating Scale (NRS), revealing the pain was due to metastatic lung cancer.

Keywords: supraclavicula brachial plexus metastasis, cervical disc herniation, brachial plexus block, metastatic lung cancer

Procedia PDF Downloads 42

Authors: Ghada E. Esheba, Ghufran Kheshaifaty, Kholoud Al-Harbi, Wafa'a Al-Harbi, Ala'a Al-Orabi, Moayad Turkistani

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Krukenberg tumor is a rare metastatic ovarian carcinoma that usually occurs in female between 30 - 40 year old and rarely seen after menopause. Stomach is the most common primary site. Histopathological features of krukenberg tumors appear as diffuse stromal proliferation, mucus-production, and numerous signet-cells and these tumors spread mostly by lymphatic route. Treatment and prognostic factors are not well established. This study describes a 34 year old female with a unilateral ovarian mass discovered accidentally during cesarean section delivery and it was misdiagnosed as luteoma of pregnancy, but histopathological examination showed a diffuse infiltration of the ovary and omentum by signet ring cells. These findings were not correlated with luteoma of pregnancy or any other types of primary ovarian tumors like surface epithelial tumor, sex cord stromal tumor or germ cell tumor. However, after the analysis of immunohistochemical results (negative CK7, positive CK20 and CDX-2), the finding was the diagnostic of metastatic krukenberg tumor. Two weeks later, the patient was evaluated and a large gastric tumor was found in her stomach and she underwent gastrectomy.

Keywords: CK7, CK20, CDX-2, Krukenburg tumor, metastatic ovarian tumor

Procedia PDF Downloads 313

Authors: G. V. Novikov, V. S. Sivozhelezov, S. S. Kolesnikov, K. V. Shaitan

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The study reports about the influence of binding of orthosteric ligands as well as point mutations on the conformational dynamics of β-2-adrenoreceptor. Using molecular dynamics simulation we found that there was a little fraction of active states of the receptor in its apo (ligand free) ensemble corresponded to its constitutive activity. Analysis of MD trajectories indicated that such spontaneous activation of the receptor is accompanied by the motion in intracellular part of its alpha-helices. Thus receptor’s constitutive activity directly results from its conformational dynamics. On the other hand the binding of a full agonist resulted in a significant shift of the initial equilibrium towards its active state. Finally, the binding of the inverse agonist stabilized the receptor in its inactive state. It is likely that the binding of inverse agonists might be a universal way of constitutive activity inhibition in vivo. Our results indicate that ligand binding redistribute pre-existing conformational degrees of freedom (in accordance to the Monod-Wyman-Changeux-Model) of the receptor rather than cause induced fit in it. Therefore, the ensemble of biologically relevant receptor conformations is encoded in its spatial structure, and individual conformations from that ensemble might be used by the cell in conformity with the physiological behaviour.

Keywords: seven-transmembrane receptors, constitutive activity, activation, x-ray crystallography, principal component analysis, molecular dynamics simulation

Procedia PDF Downloads 254

Authors: S. O. Ekpo

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Environmental science deals with everyday challenges such as a cell for call for good and safe quality air, water, food and healthy leaving condition which include destruction of biodiversity and how to conserve these natural resources for sustainable development. Biodiversity or species richness is the sum of all the different species of animals, plants, fungi and microorganisms leaving on earth and variety of habitats in which they leave. Human beings leave on plants and animals on daily basis for food, clothing, medicine, housing, research and trade or commerce; besides this, biodiversity serves to purify the air, water and land of contaminant, and recycle useful materials for continual use of man. However, man continual incessant exploitation and exploration has affected biodiversity negatively in many ways such habitant fragmentation and destruction, introduction of invasive species, pollution, overharvesting, prediction and pest control amongst others. Measures such as recycling material, establishing natural parks, sperm bank, limiting the exploitation of renewable resources to sustainable yield and urban and industrial development as well as prohibiting hunting endangered species and release of non native live forms into an area will go a long way towards conserving biodiversity for continues profitable yield.

Keywords: biodiversity, conservation, exploitation and exploration sustainable yield, recycling of materials

Procedia PDF Downloads 223

Authors: Sadia Arshad, Yifa Tang, Dumitru Baleanu

Abstract:

A fractional order mathematical model is proposed that incorporate CD8+ cells, natural killer cells, cytokines and tumor cells. The tumor cells growth in the absence of an immune response is modeled by logistic law as it was the simplest form for which predictions also agreed with the experimental data. Natural Killer Cells are our first line of defense. NK cells directly kill tumor cells through several mechanisms, including the release of cytoplasmic granules containing perforin and granzyme, expression of tumor necrosis factor (TNF) family members. The effect of the NK cells on the tumor cell population is expressed with the product term. Rational form is used to describe interaction between CD8+ cells and tumor cells. A number of cytokines are produced by NKs, including tumor necrosis factor TNF, IFN, and interleukin (IL-10). Source term for cytokines is modeled by Michaelis-Menten form to indicate the saturated effects of the immune response. Stability of the equilibrium points is discussed for biologically significant values of bifurcation parameters. We studied the treatment of fractional order system by investigating analytical conditions of tumor eradication. Numerical simulations are presented to illustrate the analytical results.

Keywords: cancer model, fractional calculus, numerical simulations, stability analysis

Procedia PDF Downloads 313

Authors: Eun Young Choi, So Hui Choe, Jin Yi Hyeon, Ji Young Jin, Bo Ram Keum, Jong Min Lim, Hyung Rae Cho, Kwang Keun Cho, In Soon Choi

Abstract:

This research analyzed the effect of β-glucan that is expected to alleviate the production of inflammatory mediator in macrophagocyte, which was processed by the lipopolysaccharide (LPS) of Escherichia, a pathogen related to allergy. The incubated layer was used for nitric oxide (NO) analysis. The DNA-binding activation of the small unit of NF-κB was measured using ELISA-based kit. In RAW264.7 cells that were vitalized by E.coli LPS, β-glucan inhibited both the combatant and rendering phases of inducible NO synthase (iNOS)-derived NO. β-glucan increased the expression of heme oxygenase-1 (HO-1) in the cell that was stimulated by E.coli LPS, and HO-1 activation was inhibited by SnPP. This shows that NO production induced by LPS is related to the inhibition effect of β-glucan. The phosphorylation of JNK and p38 induced by LPS were not influenced by β-glucan, and IκB-α decomposition was not influenced either. Instead, β-glucan remarkably inhibited the phosphorylation of STAT1 that was induced by E.coli LPS. Overall, β-glucan inhibited the production of NO in macrophagocyte that was vitalized by E.coli LPS through HO-1 induction and STAT1 pathways inhibition in this research. As the host inflammation reaction control by β-glucan weakens the progress of allergy, β-glucan can be used as an effective treatment method.

Keywords: β-glucan, lipopolysaccharide (LPS), nitric oxide (NO), RAW264.7 cells, STAT1

Procedia PDF Downloads 406