Search results for: heterogeneous cells
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 3830

Search results for: heterogeneous cells

2390 Visualization of Interaction between Pochonia Chlamydosporia and Meloidogyne Incognita and Their Impact on Tomato Crop

Authors: Saifullah K., Muhammad Naziruddin Saifullah, Muhammad N.

Abstract:

The bio control potential and mechanism of P. chlamydosporia against Meloidogyne incognita was evaluated in the present study. Under invitro conditions, P. chlamydosporia was tested for parasitism of eggs and females of M. incognita. The results indicated that this fungus parasitized 87% eggs and 82% females. Culture filtrate (CF) of P. chlamydosporia was tested for its larvicide activity against M. incognita 2nd stage juvenile. The maximum mortality was 97.3% at 100% concentration of the culture filtrate while minimum mortality was 7.3% in 25% concentration after 24 hrs. The result of the pot experiment proved that P. chlamydosporia has reduced the incidence of RKN and improved all tested agronomic growth parameters. The treatment with inoculated M. incognita alone reduced plant height, fresh shoot, and fresh root weight by 44.7%, 29.8%, and 32.8% respectively over uninoculated healthy control. Histopathological studies on the interaction of Pochonia chlamydosporia and Meloidogyne incognita on tomato roots revealed anatomical changes among treatments. Less number of galls with small in size and scarcer abnormalities in the vascular cylinder was observed in plants inoculated with P. chlamydosporia and M. incognita than the plants treated with nematode only. The fungus was seen in in the intercellular spaces of cortical and epidermal cells while the vascular bundles of the plant remain intact, inoculated only with P. chlamydosporia. In the infected roots, many mature females were seen which feed on giant cells. The findings also revealed that control healthy plants were not affected and no histological changes were noted.

Keywords: histopathology, Pochonia chlamydosporia, Meloidogyne incognita, tomato

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2389 Influence of Nitrogen Doping on the Catalytic Activity of Ni-Incorporated Carbon Nanofibers for Alkaline Direct Methanol Fuel Cells

Authors: Mohamed H. El-Newehy, Badr M. Thamer, Nasser A. M. Barakat, Mohammad A.Abdelkareem, Salem S. Al-Deyab, Hak Y. Kim

Abstract:

In this study, the influence of nitrogen doping on the electrocatalytic activity of carbon nanofibers with nickel nanoparticles toward methanol oxidation is introduced. The modified carbon nanofibers have been synthesized from calcination of electrospun nanofiber mats composed of nickel acetate tetrahydrate, poly(vinyl alcohol) and urea in argon atmosphere at 750oC. The utilized physicochemical characterizations indicated that the proposed strategy leads to form carbon nanofibers having nickel nanoparticles and doped by nitrogen. Moreover, due to the high-applied voltage during the electrospinning process, the utilized urea chemically bonds with the polymer matrix, which leads to form nitrogen-doped CNFs after the calcination process. Investigation of the electrocatalytic activity indicated that nitrogen doping NiCNFs strongly enhances the oxidation process of methanol as the current density increases from 52.5 to 198.5 mA/cm2 when the urea content in the original electrospun solution was 4 wt% urea. Moreover, the nanofibrous morphology exhibits distinct impact on the electrocatalytic activity. Also, nitrogen-doping enhanced the stability of the introduced Ni-based electrocatalyst. Overall, the present study introduces effective and simple strategy to modify the electrocatalytic activity of the nickel-based materials.

Keywords: electrospinning, methanol electrooxidation, fuel cells, nitrogen-doping, nickel

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2388 Synthesis and Evaluation of Heterogeneous Nano-Catalyst: Cr Loaded in to MCM-41

Authors: A. Salemi Golezania, A. Sharifi Fateha

Abstract:

In this study a nano-composite catalyst was synthesized by incorporation of chromium into the framework of MCM-41 as a base catalyst. Mesoporous silica molecular sieves MCM-41 were synthesized under Hydrothermal Continues pH Adjusting Path Way. Then, MCM-41 was impregnated by chromium nitrate aqueous solution for several times under water aspiration. Raw powder was cured by heat treatment in vacuum furnace at 500°C. Phase formation, morphology and gas absorption properties of resulted materials were characterized by XRD, TEM and BET analysis, respectively. The results showed that high quality hexagonal meso structure as a matrix and Cr as a second phase has been formed with a narrow size pore diameter distribution and high surface area in Cr/MCM-41 nano-composite structure. The specific surface and total volume of porosity of the synthesized nanocomposite are obtained 931m^2/gr and 1.12 cm^3/gr, respectively.

Keywords: nano-catalyst, MCM-41, Cr/MCM-41, Marine Science and Engineering

Procedia PDF Downloads 381
2387 Unified Structured Process for Health Analytics

Authors: Supunmali Ahangama, Danny Chiang Choon Poo

Abstract:

Health analytics (HA) is used in healthcare systems for effective decision-making, management, and planning of healthcare and related activities. However, user resistance, the unique position of medical data content, and structure (including heterogeneous and unstructured data) and impromptu HA projects have held up the progress in HA applications. Notably, the accuracy of outcomes depends on the skills and the domain knowledge of the data analyst working on the healthcare data. The success of HA depends on having a sound process model, effective project management and availability of supporting tools. Thus, to overcome these challenges through an effective process model, we propose an HA process model with features from the rational unified process (RUP) model and agile methodology.

Keywords: agile methodology, health analytics, unified process model, UML

Procedia PDF Downloads 500
2386 Anti-Osteoporotic Effect of Deer Antler in Ovariectomized Rats

Authors: Hye Kyung Kim, Myung-Gyou Kim, Kang-Hyun Leem

Abstract:

The deer velvet antler is well known for its traditional medicinal value and is widely used in the clinic. It has been considered to possess bone-strengthening activity. The goal of this study was to investigate the anti-osteoporotic effect of deer antler velvet on ovariectomized rats (OVX), and their possible mechanism of the action. In the first step, the in vitro effects of DAE on bone loss were determined. The proliferation, collagen content and alkaline phosphatase (ALP) activity of human osteoblastic MG-63 cells and osteoclastogenesis from bone marrow-derived precursor cells were measured. The in vivo experiment confirmed the positive effect of DAE on bone tissue. 3-month old female Sparague-Dawley rats were either sham operated or OVX, and administered DAE (20 and 100 mg/kg) for 4 weeks. DAE increased MG-63 cell proliferation and ALP activity in a dose-dependent manner. Collagen content was also increased by DAE treatment. However, the effect of DAE on bone resorption was not observed. OVX rats supplemented with DAE showed osteoprotective effects as the bone ALP level was increased and c-terminal telopeptide level was decreased by 100 mg/kg DAE treatment compared with OVX controls. Moreover, the tartrate-resistant acid phosphatase-5b level was also decreased by DAE treatment. The present study suggests that DAE is effective in preventing bone loss in OVX rats, and may be potential therapeutic agents for the treatment of postmenopausal osteoporosis.

Keywords: bone ALP, c-terminal telopeptide, deer antler, osteoporosis, ovariectomy, tartrate-resistant acid phosphatase-5b

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2385 Fucoidan: A Potent Seaweed-Derived Polysaccharide with Immunomodulatory and Anti-inflammatory Properties

Authors: Tauseef Ahmad, Muhammad Ishaq, Mathew Eapen, Ahyoung Park, Sam Karpiniec, Vanni Caruso, Rajaraman Eri

Abstract:

Fucoidans are complex, fucose-rich sulfated polymers discovered in brown seaweeds. Fucoidans are popular around the world, particularly in the nutraceutical and pharmaceutical industries, due to their promising medicinal properties. Fucoidans have been shown to have a variety of biological activities, including anti-inflammatory effects. They are known to inhibit inflammatory processes through a variety of mechanisms, including enzyme inhibition and selectin blockade. Inflammation is a part of the complicated biological response of living systems to damaging stimuli, and it plays a role in the pathogenesis of a variety of disorders, including arthritis, inflammatory bowel disease, cancer, and allergies. In the current investigation, various fucoidan extracts from Undaria pinnatifida, Fucus vesiculosus, Macrocystis pyrifera, Ascophyllum nodosum, and Laminaria japonica were assessed for inhibition of pro-inflammatory cytokine production (TNF-α, IL-1β, and IL-6) in LPS induced human macrophage cell line (THP-1) and human peripheral blood mononuclear cells (PBMCs). Furthermore, we also sought to catalogue these extracts based on their anti-inflammatory effects in the current in-vitro cell model. Materials and Methods: To assess the cytotoxicity of fucoidan extracts, MTT (3-[4,5-dimethylthiazol-2-yl]-2,5, -diphenyltetrazolium bromide) cell viability assay was performed. Furthermore, a dose-response for fucoidan extracts was performed in LPS induced THP-1 cells and PBMCs after pre-treatment for 24 hours, and levels of TNF-α, IL-1β, and IL-6 cytokines were measured using Enzyme-Linked Immunosorbent Assay (ELISA). Results: The MTT cell viability assay demonstrated that fucoidan extracts exhibited no evidence of cytotoxicity in THP-1 cells or PBMCs after 48 hours of incubation. The results of the sandwich ELISA revealed that all fucoidan extracts suppressed cytokine production in LPS-stimulated PBMCs and human THP-1 cells in a dose-dependent manner. Notably, at lower concentrations, the lower molecular fucoidan (5-30 kDa) extract from Macrocystis pyrifera was a highly efficient inhibitor of pro-inflammatory cytokines. Fucoidan extracts from all species including Undaria pinnatifida, Fucus vesiculosus, Macrocystis pyrifera, Ascophyllum nodosum, and Laminaria japonica exhibited significant anti-inflammatory effects. These findings on several fucoidan extracts provide insight into strategies for improving their efficacy against inflammation-related diseases. Conclusion: In the current research, we have successfully catalogued several fucoidan extracts based on their efficiency in LPS-induced macrophages and PBMCs in downregulating the key pro-inflammatory cytokines (TNF-, IL-1 and IL-6), which are prospective targets in human inflammatory illnesses. Further research would provide more information on the mechanism of action, allowing it to be tested for therapeutic purposes as an anti-inflammatory medication.

Keywords: fucoidan, PBMCs, THP-1, TNF-α, IL-1β, IL-6, inflammation

Procedia PDF Downloads 57
2384 Kirchhoff’s Depth Migration over Heterogeneous Velocity Models with Ray Tracing Modeling Approach

Authors: Alok Kumar Routa, Priya Ranjan Mohanty

Abstract:

Complex seismic signatures are generated due to the complexity of the subsurface which is difficult to interpret. In the present study, an attempt has been made to model the complex subsurface using the Ray tracing modeling technique. Add to this, for the imaging of these geological features, Kirchhoff’s prestack depth migration is applied over the synthetic common shot gather dataset. It is found that the Kirchhoff’s migration technique in addition with the Ray tracing modeling concept has the flexibility towards the imaging of various complex geology which gives satisfactory results with proper delineation of the reflectors at their respective true depth position. The entire work has been carried out under the MATLAB environment.

Keywords: Kirchhoff's migration, Prestack depth migration, Ray tracing modelling, velocity model

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2383 Use of 3D Printed Bioscaffolds from Decellularized Umbilical Cord for Cartilage Regeneration

Authors: Tayyaba Bari, Muhammad Hamza Anjum, Samra Kanwal, Fakhera Ikram

Abstract:

Osteoarthritis, a degenerative condition, affects more than 213 million individuals globally. Since articular cartilage has no or limited vessels, therefore, after deteriorating, it is unable to rejuvenate. Traditional approaches for cartilage repair, like autologous chondrocyte implantation, microfracture and cartilage transplantation are often associated with postoperative complications and lead to further degradation. Decellularized human umbilical cord has gained interest as a viable treatment for cartilage repair. Decellularization removes all cellular contents as well as debris, leaving a biologically active 3D network known as extracellular matrix (ECM). This matrix is biodegradable, non-immunogenic and provides a microenvironment for homeostasis, growth and repair. UC derived bioink function as 3D scaffolding material, not only mediates cell-matrix interactions but also adherence, proliferation and propagation of cells for 3D organoids. This study comprises different physical, chemical and biological approaches to optimize the decellularization of human umbilical cord (UC) tissues followed by the solubilization of these tissues to bioink formation. The decellularization process consisted of two cycles of freeze thaw where the umbilical cord at -20˚C was thawed at room temperature followed by dissection in small sections from 0.5 to 1cm. Similarly decellularization with ionic and non-ionic detergents Sodium dodecyl sulfate (SDS) and Triton-X 100 revealed that both concentrations of SDS i.e 0.1% and 1% were effective in complete removal of cells from the small UC tissues. The results of decellularization was further confirmed by running them on 1% agarose gel. Histological analysis revealed the efficacy of decellularization, which involves paraffin embedded samples of 4μm processed for Hematoxylin-eosin-safran and 4,6-diamidino-2-phenylindole (DAPI). ECM preservation was confirmed by Alcian Blue, and Masson’s trichrome staining on consecutive sections and images were obtained. Sulfated GAG’s content were determined by 1,9-dimethyl-methylene blue (DMMB) assay, similarly collagen quantification was done by hydroxy proline assay. This 3D bioengineered scaffold will provide a typical atmosphere as in the extracellular matrix of the tissue, which would be seeded with the mesenchymal cells to generate the desired 3D ink for in vitro and in vivo cartilage regeneration applications.

Keywords: umbilical cord, 3d printing, bioink, tissue engineering, cartilage regeneration

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2382 Optimized Approach for Secure Data Sharing in Distributed Database

Authors: Ahmed Mateen, Zhu Qingsheng, Ahmad Bilal

Abstract:

In the current age of technology, information is the most precious asset of a company. Today, companies have a large amount of data. As the data become larger, access to data for some particular information is becoming slower day by day. Faster data processing to shape it in the form of information is the biggest issue. The major problems in distributed databases are the efficiency of data distribution and response time of data distribution. The security of data distribution is also a big issue. For these problems, we proposed a strategy that can maximize the efficiency of data distribution and also increase its response time. This technique gives better results for secure data distribution from multiple heterogeneous sources. The newly proposed technique facilitates the companies for secure data sharing efficiently and quickly.

Keywords: ER-schema, electronic record, P2P framework, API, query formulation

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2381 Stability Analysis of Two-delay Differential Equation for Parkinson's Disease Models with Positive Feedback

Authors: M. A. Sohaly, M. A. Elfouly

Abstract:

Parkinson's disease (PD) is a heterogeneous movement disorder that often appears in the elderly. PD is induced by a loss of dopamine secretion. Some drugs increase the secretion of dopamine. In this paper, we will simply study the stability of PD models as a nonlinear delay differential equation. After a period of taking drugs, these act as positive feedback and increase the tremors of patients, and then, the differential equation has positive coefficients and the system is unstable under these conditions. We will present a set of suggested modifications to make the system more compatible with the biodynamic system. When giving a set of numerical examples, this research paper is concerned with the mathematical analysis, and no clinical data have been used.

Keywords: Parkinson's disease, stability, simulation, two delay differential equation

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2380 Effects of Punicalagin on Some Productive and Reproductive Traits in Virgin Rabbit Does

Authors: Nada A. El-Shahaw, Anas A.Salem, M. Kobeisy, Hoda M. Shabaan

Abstract:

Reactive oxygen species (ROS) is collective term both oxygen radical, such superoxide (O₂•), hydroxyl(OH•), peroxyl (RO₂), and hydroperoxyl (HO₂•), and certain non-radical oxidizing agents, such as hydrogen peroxide (H₂O₂), hypochlorous acid (HOCL), and ozone (O₃), that can be convert easily to radical. The importance of antioxidants is shown here punicalagin. Punicalagin is preventing the harmful effect of (ROS) in all cells, specially gonadal cells. So, the aim of study was to investigate effects of punicalagin (PL) on maternal live body weight (MLBW), number of services/conception (NS), conception rate (CR), gestation length (GL), kindling rate (KR), total litter size (TLS), live litter size (LLS), kit weight (KW), progesterone (P4) and estradiol-17 (E2) concentrations at 1st and 2nd pregnancy of young does. A total of 28 healthy virgin does (6 months old) were divided into 2 equal groups. Group I, each doe, was injected IM with 100 ug PL twice/week pre-mating and one time 3 days post-mating. Group II, each doe was injected IM with sterilized water (control). Blood samples were taken at pre-mating, mating, post-mating, throughout pregnancy, and immediately post-kindling for assaying P4 and E2. All does were naturally mated with fertile bucks. Results revealed that PL displayed their significant impacts on MLBW, NS/conception, CR, GL, KR, TLS, LLS, KWs (birth and weaning), P4 and E2 concentrations either at 1ˢᵗ/2ⁿᵈ pregnancy or both of them. Conclusively, PL improved pregnancy outcomes of young do particularly at 2ⁿᵈ pregnancy and could be recommended in rabbit's farms.

Keywords: punicalagin, pregnancy, estradiol-17β, progesterone, does

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2379 Cardiotoxicity Associated with Radiation Therapy: The Role of Bone Marrow Mesenchymal Cells in Improvement of Heart Function

Authors: Isalira Peroba Ramos, Cherley Borba Vieira de Andrade, Grazielle Suhett, Camila Salata, Paulo Cesar Canary, Guilherme Visconde Brasil, Antonio Carlos Campos de Carvalho, Regina Coeli dos Santos Goldenberg

Abstract:

Background: The therapeutic options for patients with cancer now include increasingly complex combinations of medications, radiation therapy (RT), and surgical intervention. Many of these treatments have important potential adverse cardiac effects and are likely to have significant effects on patient outcomes. Cell therapy appears to be promising for the treatment of chronic and degenerative diseases, including cardiomyopathy induced by RT, as the current therapeutic options are insufficient. Aims: To evaluate the potential of bone marrow mesenchymal cells (BMMCs) in radioinduced cardiac damage Methods: Female Wistar rats, 3 months old (Ethics Committee 054/14), were divided into 2 groups, non-treated irradiated group (IR n=15) and irradiated and BMMC treated (IRT n=10). Echocardiography was performed to evaluate heart function. After euthanasia, 3 months post treatment; the left ventricle was removed and prepared for RT-qPCR (VEGF and Pro Collagen I) and histological (picrosirius) analysis. Results: In both groups, 45 days after irradiation, ejection fraction (EF) was in the normal range for these animals (> 70%). However, the BMMC treated group had EF (83.1%±2.6) while the non-treated IR group showed a significant reduction (76.1%±2.6) in relation to the treated group. In addition, we observed an increase in VEGF gene expression and a decrease in Pro Collagen I in IRT when compared to IR group. We also observed by histology that the collagen deposition was reduced in IRT (10.26%±0.83) when compared to IR group (25.29%±0.96). Conclusions: Treatment with BMMCs was able to prevent ejection fraction reduction and collagen deposition in irradiated animals. The increase of VEGF and the decrease of pro collagen I gene expression might explain, at least in part, the cell therapy benefits. All authors disclose no financial or personal relationships with individuals or organizations that could be perceived to bias their work. Sources of funding: FAPERJ, CAPES, CNPq, MCT.

Keywords: mesenchymal cells, radioation, cardiotoxicity, bone marrow

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2378 Efficient Production of Cell-Adhesive Motif From Human Fibronectin Domains to Design a Bio-Functionalized Scaffold for Tissue Engineering

Authors: Amina Ben Abla, Sylvie Changotade, Geraldine Rohman, Guilhem Boeuf, Cyrine Dridi, Ahmed Elmarjou, Florence Dufour, Didier Lutomski, Abdellatif Elm’semi

Abstract:

Understanding cell adhesion and interaction with the extracellular matrix is essential for biomedical and biotechnological applications, including the development of biomaterials. In recent years, numerous biomaterials have emerged and were used in the field of tissue engineering. Nevertheless, the lack of interaction of biomaterials with cells still limits their bio-integration. Thus, the design of bioactive biomaterials to improve cell attachment and proliferation is of growing interest. In this study, bio-functionalized material was developed combining a synthetic polymer scaffold surface with selected domains of type III human fibronectin (FNIII-DOM) to promote cell adhesion and proliferation. Bioadhesive ligand includes cell-binding domains of human fibronectin, a major ECM protein that interacts with a variety of integrins cell-surface receptors, and ECM proteins through specific binding domains were engineered. FNIII-DOM was produced in bacterial system E. coli in 5L fermentor with a high yield level reaching 20mg/L. Bioactivity of the produced fragment was validated by studying cellular adhesion of human cells. The adsorption and immobilization of FNIII-DOM onto the polymer scaffold were evaluated in order to develop an innovative biomaterial.

Keywords: biomaterials, cellular adhesion, fibronectin, tissue engineering

Procedia PDF Downloads 139
2377 Functional Profiling of a Circular RNA from the Huntingtin (HTT) Gene

Authors: Laura Gantley, Vanessa M. Conn, Stuart Webb, Kirsty Kirk, Marta Gabryelska, Duncan Holds, Brett W. Stringer, Simon J. Conn

Abstract:

Trinucleotide repeat disorders comprise ~20 severe, inherited human neuromuscular and neurodegenerative disorders, which are a result of an abnormal expansion of repetitive sequences in the DNA. The most common of these, Huntington’s disease, results from the expansion of the CAG repeat region in exon 1 of the HTT gene via an unknown mechanism. Non-coding RNAs have been implicated in the initiation and progression of many diseases; thus, we focus on one circular RNA (circRNA) molecule arising from non-canonical splicing (back splicing) of HTT pre-mRNA. This circRNA and its mouse orthologue were transgenically overexpressed in human cells (SHSY-5Y and HEK293T) and mouse cells (Mb1), respectively. High-content imaging and flow cytometry demonstrated the overexpression of this circRNA reduces cell proliferation, reduces nuclear size independent of cellular size, and alters cell cycle progression. Analysis of protein by western blot and immunofluorescence demonstrated no change to HTT protein levels but altered nuclear-cytoplasmic distribution without impacting the expansion of the HTT repeat region. As these phenotypic and genotypic changes are found in Huntington’s disease patients, these results may suggest that this circRNA may play a functional role in the progression of Huntington’s disease.

Keywords: cell biology, circular RNAs, Huntington’s disease, molecular biology, neurodegenerative disorders

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2376 Biodegradation of Phenazine-1-Carboxylic Acid by Rhodanobacter sp. PCA2 Proceeds via Decarboxylation and Cleavage of Nitrogen-Containing Ring

Authors: Miaomiao Zhang, Sabrina Beckmann, Haluk Ertan, Rocky Chau, Mike Manefield

Abstract:

Phenazines are a large class of nitrogen-containing aromatic heterocyclic compounds, which are almost exclusively produced by bacteria from diverse genera including Pseudomonas and Streptomyces. Phenazine-1-carboxylic acid (PCA) as one of 'core' phenazines are converted from chorismic acid before modified to other phenazine derivatives in different cells. Phenazines have attracted enormous interests because of their multiple roles on biocontrol, bacterial interaction, biofilm formation and fitness of their producers. However, in spite of ecological importance, degradation as a part of phenazines’ fate only have extremely limited attention now. Here, to isolate PCA-degrading bacteria, 200 mg L-1 PCA was supplied as sole carbon, nitrogen and energy source in minimal mineral medium. Quantitative PCR and Reverse-transcript PCR were employed to study abundance and activity of functional gene MFORT 16269 in PCA degradation, respectively. Intermediates and products of PCA degradation were identified with LC-MS/MS. After enrichment and isolation, a PCA-degrading strain was selected from soil and was designated as Rhodanobacter sp. PCA2 based on full 16S rRNA sequencing. As determined by HPLC, strain PCA2 consumed 200 mg L-1 (836 µM) PCA at a rate of 17.4 µM h-1, accompanying with significant cells yield from 1.92 × 105 to 3.11 × 106 cells per mL. Strain PCA2 was capable of degrading other phenazines as well, including phenazine (4.27 µM h-1), pyocyanin (2.72 µM h-1), neutral red (1.30 µM h-1) and 1-hydroxyphenazine (0.55 µM h-1). Moreover, during the incubation, transcript copies of MFORT 16269 gene increased significantly from 2.13 × 106 to 8.82 × 107 copies mL-1, which was 2.77 times faster than that of the corresponding gene copy number (2.20 × 106 to 3.32 × 107 copies mL-1), indicating that MFORT 16269 gene was activated and played roles on PCA degradation. As analyzed by LC-MS/MS, decarboxylation from the ring structure was determined as the first step of PCA degradation, followed by cleavage of nitrogen-containing ring by dioxygenase which catalyzed phenazine to nitrosobenzene. Subsequently, phenylhydroxylamine was detected after incubation for two days and was then transferred to aniline and catechol. Additionally, genomic and proteomic analyses were also carried out for strain PCA2. Overall, the findings presented here showed that a newly isolated strain Rhodanobacter sp. PCA2 was capable of degrading phenazines through decarboxylation and cleavage of nitrogen-containing ring, during which MFORT 16269 gene was activated and played important roles.

Keywords: decarboxylation, MFORT16269 gene, phenazine-1-carboxylic acid degradation, Rhodanobacter sp. PCA2

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2375 IL6/PI3K/mTOR/GFAP Molecular Pathway Role in COVID-19-Induced Neurodegenerative Autophagy, Impacts and Relatives

Authors: Mohammadjavad Sotoudeheian

Abstract:

COVID-19, which began in December 2019, uses the angiotensin-converting enzyme 2 (ACE2) receptor to enter and spread through the cells. ACE2 mRNA is present in almost every organ, including nasopharynx, lung, as well as the brain. Ports of entry of SARS-CoV-2 into the central nervous system (CNS) may include arterial circulation, while viremia is remarkable. However, it is imperious to develop neurological symptoms evaluation CSF analysis in patients with COVID-19, but theoretically, ACE2 receptors are expressed in cerebellar cells and may be a target for SARS-CoV-2 infection in the brain. Recent evidence agrees that SARS-CoV-2 can impact the brain through direct and indirect injury. Two biomarkers for CNS injury, glial fibrillary acidic protein (GFAP) and neurofilament light chain (NFL) detected in the plasma of patients with COVID-19. NFL, an axonal protein expressed in neurons, is related to axonal neurodegeneration, and GFAP is over-expressed in CNS inflammation. GFAP cytoplasmic accumulation causes Schwan cells to misfunction, so affects myelin generation, reduces neuroskeletal support over NfLs during CNS inflammation, and leads to axonal degeneration. Interleukin-6 (IL-6), which extensively over-express due to interleukin storm during COVID-19 inflammation, regulates gene expression, as well as GFAP through STAT molecular pathway. IL-6 also impresses the phosphoinositide 3-kinase (PI3K)/STAT/smads pathway. The PI3K/ protein kinase B (Akt) pathway is the main modulator upstream of the mammalian target of rapamycin (mTOR), and alterations in this pathway are common in neurodegenerative diseases. Most neurodegenerative diseases show a disruption of autophagic function and display an abnormal increase in protein aggregation that promotes cellular death. Therefore, induction of autophagy has been recommended as a rational approach to help neurons clear abnormal protein aggregates and survive. The mTOR is a major regulator of the autophagic process and is regulated by cellular stressors. The mTORC1 pathway and mTORC2, as complementary and important elements in mTORC1 signaling, have become relevant in the regulation of the autophagic process and cellular survival through the extracellular signal-regulated kinase (ERK) pathway.

Keywords: mTORC1, COVID-19, PI3K, autophagy, neurodegeneration

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2374 In vitro Comparison Study of Biologically Synthesized Cupper-Disulfiram Nanoparticles with Its Free Corresponding Complex as Therapeutic Approach for Breast and Liver Cancer

Authors: Marwa M. Abu-Serie, Marwa M. Eltarahony

Abstract:

The search for reliable, effective, and safe nanoparticles (NPs) as a treatment for cancer is a pressing priority. In this study, Cu-NPs were fabricated by Streptomyces cyaneofuscatus through simultaneous bioreduction strategy of copper nitrate salt. The as-prepared Cu-NPs subjected to structural analysis; energy-dispersive X-ray spectroscopy, elemental mapping, X-ray diffraction, transmission electron microscopy, and ζ-potential. These biological synthesized Cu-NPs were mixed with disulfiram (DS), forming a nanocomplex of Cu-DS with a size of ~135 nm. The prepared nanocomplex (nanoCu-DS) exhibited higher anticancer activity than that of free complex of DS-Cu, Cu-NPs, and DS alone. This was illustrated by the lowest IC50 of nanoCu-DS (< 4 µM) against human breast and liver cancer cell lines comparing with DS-Cu, Cu-NPs, and DS (~8, 22.98-33.51 and 11.95-14.86, respectively). Moreover, flow cytometric analysis confirmed that higher apoptosis percentage range of nanoCu-DS-treated in MDA-MB 231, MCF-7, Huh-7, and HepG-2 cells (51.24-65.28%) than free complex of Cu-DS ( < 4.5%). Regarding inhibition potency of liver and breast cancer cell migration, no significant difference was recorded between free and nanocomplex. Furthermore, nanoCu-DS suppressed gene expression of β-catenine, Akt, and NF-κB and upregulated p53 expression (> 3, >15, > 5 and ≥ 3 folds, respectively) more efficiently than free complex (all ~ 1 fold) in MDA-MB 231 and Huh-7 cells. Our finding proved this prepared nano complex has a powerful anticancer activity relative to free complex, thereby offering a promising cancer treatment.

Keywords: biologically prepared Cu-NPs, breast cancer cell lines, liver cancer cell lines, nanoCu- disulfiram

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2373 Development of Three-Dimensional Bio-Reactor Using Magnetic Field Stimulation to Enhance PC12 Cell Axonal Extension

Authors: Eiji Nakamachi, Ryota Sakiyama, Koji Yamamoto, Yusuke Morita, Hidetoshi Sakamoto

Abstract:

The regeneration of injured central nerve network caused by the cerebrovascular accidents is difficult, because of poor regeneration capability of central nerve system composed of the brain and the spinal cord. Recently, new regeneration methods such as transplant of nerve cells and supply of nerve nutritional factor were proposed and examined. However, there still remain many problems with the canceration of engrafted cells and so on and it is strongly required to establish an efficacious treating method of a central nerve system. Blackman proposed the electromagnetic stimulation method to enhance the axonal nerve extension. In this study, we try to design and fabricate a new three-dimensional (3D) bio-reactor, which can load a uniform AC magnetic field stimulation on PC12 cells in the extracellular environment for enhancement of an axonal nerve extension and 3D nerve network generation. Simultaneously, we measure the morphology of PC12 cell bodies, axons, and dendrites by the multiphoton excitation fluorescence microscope (MPM) and evaluate the effectiveness of the uniform AC magnetic stimulation to enhance the axonal nerve extension. Firstly, we designed and fabricated the uniform AC magnetic field stimulation bio-reactor. For the AC magnetic stimulation system, we used the laminated silicon steel sheets for a yoke structure of 3D chamber, which had a high magnetic permeability. Next, we adopted the pole piece structure and installed similar specification coils on both sides of the yoke. We searched an optimum pole piece structure using the magnetic field finite element (FE) analyses and the response surface methodology. We confirmed that the optimum 3D chamber structure showed a uniform magnetic flux density in the PC12 cell culture area by using FE analysis. Then, we fabricated the uniform AC magnetic field stimulation bio-reactor by adopting analytically determined specifications, such as the size of chamber and electromagnetic conditions. We confirmed that measurement results of magnetic field in the chamber showed a good agreement with FE results. Secondly, we fabricated a dish, which set inside the uniform AC magnetic field stimulation of bio-reactor. PC12 cells were disseminated with collagen gel and could be 3D cultured in the dish. The collagen gel were poured in the dish. The collagen gel, which had a disk shape of 6 mm diameter and 3mm height, was set on the membrane filter, which was located at 4 mm height from the bottom of dish. The disk was full filled with the culture medium inside the dish. Finally, we evaluated the effectiveness of the uniform AC magnetic field stimulation to enhance the nurve axonal extension. We confirmed that a 6.8 increase in the average axonal extension length of PC12 under the uniform AC magnetic field stimulation at 7 days culture in our bio-reactor, and a 24.7 increase in the maximum axonal extension length. Further, we confirmed that a 60 increase in the number of dendrites of PC12 under the uniform AC magnetic field stimulation. Finally, we confirm the availability of our uniform AC magnetic stimulation bio-reactor for the nerve axonal extension and the nerve network generation.

Keywords: nerve regeneration, axonal extension , PC12 cell, magnetic field, three-dimensional bio-reactor

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2372 The Effect of High-Pressure Processing on the Inactivation of Saccharomyces cerevisiae in Different Concentration of Manuka Honey and Its Relation with ° Brix

Authors: Noor Akhmazillah Fauzi, Mohammed Mehdi Farid, Filipa V. Silva

Abstract:

The aim of this paper is to investigate if different concentration of Manuka honey (as a model food) has a major influence on the inactivation of Saccharomyces cerevisiae (as the testing microorganism) after subjecting it to HPP. Honey samples with different sugar concentrations (20, 30, 40, 50, 60 and 70 °Brix) were prepared aseptically using sterilized distilled water. No dilution of honey was made for the 80 °Brix sample. For the 0 °Brix sample (control), sterilized distilled water was used. Thermal treatment at 55 °C for 10 min (conventionally applied in honey pasteurisation in industry) was carried out for comparison purpose. S. cerevisiae cell numbers in honey samples were established before and after each HPP and thermal treatment. The number of surviving cells was determined after a proper dilution of the untreated and treated samples by the viable plate count method. S. cerevisiae cells, in different honey concentrations (0 to 80 °Brix), subjected to 600 MPa (at ambient temperature) showed an increasing resistance to inactivation with °Brix. A significant correlation (p < 0.05) between cell reduction and °Brix was found. Cell reduction in high pressure-treated samples varied linearly with °Brix (R2 > 0.9), confirming that the baroprotective effect of the food is due to sugar content. This study has practical implications in establishing efficient process design for commercial manufacturing of high sugar food products and on the potential use of HPP for such products.

Keywords: high pressure processing, honey, Saccharomyces cerevisiae, °Brix

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2371 Efficient Delivery of Biomaterials into Living Organism by Using Noble Metal Nanowire Injector

Authors: Kkochorong Park, Keun Cheon Kim, Hyoban Lee, Eun Ju Lee, Bongsoo Kim

Abstract:

Introduction of biomaterials such as DNA, RNA, proteins is important for many research areas. There are many methods to introduce biomaterials into living organisms like tissue and cells. To introduce biomaterials, several indirect methods including virus‐mediated delivery, chemical reagent (i.e., lipofectamine), electrophoresis have been used. Such methods are passive delivery using an endocytosis process of cell, reducing an efficiency of delivery. Unlike the indirect delivery method, it has been reported that a direct delivery of exogenous biomolecules into nucleus have been more efficient to expression or integration of biomolecules. Nano-sized material is beneficial for detect signal from cell or deliver stimuli/materials into the cell at cellular and molecular levels, due to its similar physical scale. Especially, because 1 dimensional (1D) nanomaterials such as nanotube, nanorod and nanowire with high‐aspect ratio have nanoscale geometry and excellent mechanical, electrical, and chemical properties, they could play an important role in molecular and cellular biology. In this study, by using single crystalline 1D noble metal nanowire, we fabricated nano-sized 1D injector which can successfully interface with living cells and directly deliver biomolecules into several types of cell line (i.e., stem cell, mammalian embryo) without inducing detrimental damages on living cell. This nano-bio technology could be a promising and robust tool for introducing exogenous biomaterials into living organism.

Keywords: DNA, gene delivery, nanoinjector, nanowire

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2370 Validation of an Impedance-Based Flow Cytometry Technique for High-Throughput Nanotoxicity Screening

Authors: Melanie Ostermann, Eivind Birkeland, Ying Xue, Alexander Sauter, Mihaela R. Cimpan

Abstract:

Background: New reliable and robust techniques to assess biological effects of nanomaterials (NMs) in vitro are needed to speed up safety analysis and to identify key physicochemical parameters of NMs, which are responsible for their acute cytotoxicity. The central aim of this study was to validate and evaluate the applicability and reliability of an impedance-based flow cytometry (IFC) technique for the high-throughput screening of NMs. Methods: Eight inorganic NMs from the European Commission Joint Research Centre Repository were used: NM-302 and NM-300k (Ag: 200 nm rods and 16.7 nm spheres, respectively), NM-200 and NM- 203 (SiO₂: 18.3 nm and 24.7 nm amorphous, respectively), NM-100 and NM-101 (TiO₂: 100 nm and 6 nm anatase, respectively), and NM-110 and NM-111 (ZnO: 147 nm and 141 nm, respectively). The aim was to assess the biological effects of these materials on human monoblastoid (U937) cells. Dispersions of NMs were prepared as described in the NANOGENOTOX dispersion protocol and cells were exposed to NMs at relevant concentrations (2, 10, 20, 50, and 100 µg/mL) for 24 hrs. The change in electrical impedance was measured at 0.5, 2, 6, and 12 MHz using the IFC AmphaZ30 (Amphasys AG, Switzerland). A traditional toxicity assay, Trypan Blue Dye Exclusion assay, and dark-field microscopy were used to validate the IFC method. Results: Spherical Ag particles (NM-300K) showed the highest toxic effect on U937 cells followed by ZnO (NM-111 ≥ NM-110) particles. Silica particles were moderate to non-toxic at all used concentrations under these conditions. A higher toxic effect was seen with smaller sized TiO2 particles (NM-101) compared to their larger analogues (NM-100). No interferences between the IFC and the used NMs were seen. Uptake and internalization of NMs were observed after 24 hours exposure, confirming actual NM-cell interactions. Conclusion: Results collected with the IFC demonstrate the applicability of this method for rapid nanotoxicity assessment, which proved to be less prone to nano-related interference issues compared to some traditional toxicity assays. Furthermore, this label-free and novel technique shows good potential for up-scaling in directions of an automated high-throughput screening and for future NM toxicity assessment. This work was supported by the EC FP7 NANoREG (Grant Agreement NMP4-LA-2013-310584), the Research Council of Norway, project NorNANoREG (239199/O70), the EuroNanoMed II 'GEMN' project (246672), and the UH-Nett Vest project.

Keywords: cytotoxicity, high-throughput, impedance, nanomaterials

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2369 New Active Dioxin Response Element Sites in Regulatory Region of Human and Viral Genes

Authors: Ilya B. Tsyrlov, Dmitry Y. Oshchepkov

Abstract:

A computational search for dioxin response elements (DREs) in genes of proteins comprising the Ah receptor (AhR) cytosolic core complex was performed by highly efficient tool SITECON. Eventually, the following number of new DREs in 5’flanking region was detected by SITECON: one in AHR gene, five in XAP2, eight in HSP90AA1, and three in HSP90AB1 genes. Numerous DREs found in genes of AhR and AhR cytosolic complex members would shed a light on potential mechanisms of expression, the stoichiometry of unliganded AhR core complex, and its degradation vs biosynthesis dynamics resulted from treatment of target cells with the AhR most potent ligand, 2,3,7,8-TCDD. With human viruses, reduced susceptibility to TCDD of geneencoding HIV-1 P247 was justified by the only potential DRE determined in gag gene encoding HIV-1 P24 protein, whereas the regulatory region of CMV genes encoding IE gp/UL37 has five potent DRE, 1.65 kb/UL36 – six DRE, pp65 and pp71 – each has seven DRE, and pp150 – ten DRE. Also, from six to eight DRE were determined with SITECON in the regulatory region of HSV-1 IE genes encoding tegument proteins, UL36 and UL37, and of UL19 gene encoding bindingglycoprotein C (gC). So, TCDD in the low picomolar range may activate in human cells AhR: Arnt transcription pathway that triggers CMV and HSV-1 reactivation by binding to numerous promoter DRE within immediate-early (IE) genes UL37 and UL36, thus committing virus to the lytic cycle.

Keywords: dioxin response elements, Ah receptor, AhR: Arnt transcription pathway, human and viral genes

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2368 N-Glycosylation in the Green Microalgae Chlamydomonas reinhardtii

Authors: Pierre-Louis Lucas, Corinne Loutelier-Bourhis, Narimane Mati-Baouche, Philippe Chan Tchi-Song, Patrice Lerouge, Elodie Mathieu-Rivet, Muriel Bardor

Abstract:

N-glycosylation is a post-translational modification taking place in the Endoplasmic Reticulum and the Golgi apparatus where defined glycan features are added on protein in a very specific sequence Asn-X-Thr/Ser/Cys were X can be any amino acid except proline. Because it is well-established that those N-glycans play a critical role in protein biological activity, protein half-life and that a different N-glycan structure may induce an immune response, they are very important in Biopharmaceuticals which are mainly glycoproteins bearing N-glycans. From now, most of the biopharmaceuticals are produced by mammalian cells like Chinese Hamster Ovary cells (CHO) for their N-glycosylation similar to the human, but due to the high production costs, several other species are investigated as the possible alternative system. In this purpose, the green microalgae Chlamydomonas reinhardtii was investigated as the potential production system for Biopharmaceuticals. This choice was influenced by the facts that C. reinhardtii is a well-study microalgae which is growing fast with a lot of molecular biology tools available. This organism is also producing N-glycan on its endogenous proteins. However, the analysis of the N-glycan structure of this microalgae has revealed some differences as compared to the human. Rather than in Human where the glycans are processed by key enzymes called N-acetylglucosaminyltransferase I and II (GnTI and GnTII) adding GlcNAc residue to form a GlcNAc₂Man₃GlcNAc₂ core N-glycan, C. reinhardtii lacks those two enzymes and possess a GnTI independent glycosylation pathway. Moreover, some enzymes like xylosyltransferases and methyltransferases not present in human are supposed to act on the glycans of C. reinhardtii. Furthermore, the recent structural study by mass spectrometry shows that the N-glycosylation precursor supposed to be conserved in almost all eukaryotic cells results in a linear Man₅GlcNAc₂ rather than a branched one in C. reinhardtii. In this work, we will discuss the new released MS information upon C. reinhardtii N-glycan structure and their impact on our attempt to modify the glycan in a Human manner. Two strategies will be discussed. The first one consisted in the study of Xylosyltransferase insertional mutants from the CLIP library in order to remove xyloses from the N-glycans. The second will go further in the humanization by transforming the microalgae with the exogenous gene from Toxoplasma gondii having an activity similar to GnTI and GnTII with the aim to synthesize GlcNAc₂Man₃GlcNAc₂ in C. reinhardtii.

Keywords: Chlamydomonas reinhardtii, N-glycosylation, glycosyltransferase, mass spectrometry, humanization

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2367 Regulation of Desaturation of Fatty Acid and Triglyceride Synthesis by Myostatin through Swine-Specific MEF2C/miR222/SCD5 Pathway

Authors: Wei Xiao, Gangzhi Cai, Xingliang Qin, Hongyan Ren, Zaidong Hua, Zhe Zhu, Hongwei Xiao, Ximin Zheng, Jie Yao, Yanzhen Bi

Abstract:

Myostatin (MSTN) is the master regulator of double muscling phenotype with overgrown muscle and decreased fatness in animals, but its action mode to regulate fat deposition remains to be elucidated. In this study a swin-specific pathway through which MSTN acts to regulate the fat deposition was deciphered. Deep sequenincing of the mRNA and miRNA of fat tissues of MSTN knockout (KO) and wildtype (WT) pigs discovered the positive correlation of myocyte enhancer factor 2C (MEF2C) and fat-inhibiting miR222 expression, and the inverse correlation of miR222 and stearoyl-CoA desaturase 5 (SCD5) expression. SCD5 is rodent-absent and expressed only in pig, sheep and cattle. Fatty acid spectrum of fat tissues revealed a lower percentage of oleoyl-CoA (18:1) and palmitoleyl CoA (16:1) in MSTN KO pigs, which are the catalyzing products of SCD5-mediated desaturation of steroyl CoA (18:0) and palmitoyl CoA (16:0). Blood metrics demonstrated a 45% decline of triglyceride (TG) content in MSTN KO pigs. In light of these observations we hypothesized that MSTN might act through MEF2C/miR222/SCD5 pathway to regulate desaturation of fatty acid as well as triglyceride synthesis in pigs. To this end, real-time PCR and Western blotting were carried out to detect the expression of the three genes stated above. These experiments showed that MEF2C expression was up-regulated by nearly 2-fold, miR222 up-regulated by nearly 3-fold and SCD5 down-regulated by nearly 50% in MSTN KO pigs. These data were consistent with the expression change in deep sequencing analysis. Dual luciferase reporter was then used to confirm the regulation of MEF2C upon the promoter of miR222. Ecotopic expression of MEF2C in preadipocyte cells enhanced miR222 expression by 3.48-fold. CHIP-PCR identified a putative binding site of MEF2C on -2077 to -2066 region of miR222 promoter. Electrophoretic mobility shift assay (EMSA) demonstrated the interaction of MEF2C and miR222 promoter in vitro. These data indicated that MEF2C transcriptionally regulates the expression of miR222. Next, the regulation of miR222 on SCD5 mRNA as well as its physiological consequences were examined. Dual luciferase reporter testing revealed the translational inhibition of miR222 upon the 3´ UTR (untranslated region) of SCD5 in preadipocyte cells. Transfection of miR222 mimics and inhibitors resulted in the down-regulation and up-regulation of SCD5 in preadipocyte cells respectively, consistent with the results from reporter testing. RNA interference of SCD5 in preadipocyte cells caused 26.2% reduction of TG, in agreement with the results of TG content in MSTN KO pigs. In summary, the results above supported the existence of a molecular pathway that MSTN signals through MEF2C/miR222/SCD5 to regulate the fat deposition in pigs. This swine-specific pathway offers potential molecular markers for the development and breeding of a new pig line with optimised fatty acid composition. This would benefit human health by decreasing the takeup of saturated fatty acid.

Keywords: fat deposition, MEF2C, miR222, myostatin, SCD5, pig

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2366 Self-Organization-Based Approach for Embedded Real-Time System Design

Authors: S. S. Bendib, L. W. Mouss, S. Kalla

Abstract:

This paper proposes a self-organization-based approach for real-time systems design. The addressed issue is the mapping of an application onto an architecture of heterogeneous processors while optimizing both makespan and reliability. Since this problem is NP-hard, a heuristic algorithm is used to obtain efficiently approximate solutions. The proposed approach takes into consideration the quality as well as the diversity of solutions. Indeed, an alternate treatment of the two objectives allows to produce solutions of good quality while a self-organization approach based on the neighborhood structure is used to reorganize solutions and consequently to enhance their diversity. Produced solutions make different compromises between the makespan and the reliability giving the user the possibility to select the solution suited to his (her) needs.

Keywords: embedded real-time systems design, makespan, reliability, self-organization, compromises

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2365 The Cell Viability Study of Extracts of Bark, Flowers, Leaves and Seeds of Indian Dhak Tree, Flame of Forest

Authors: Madhavi S. Apte, Milind Bhitre

Abstract:

In pharmaceutical research and new drug development, medicinal plants have important roles. Similarly, Indian dhak tree belonging to family Fabaceae has been widely used in the traditional Indian medical system of ‘Ayurveda’ for the treatment of a variety of ailments. Hence the cell viability study was undertaken to evaluate and compare the activity of extracts of various parts like flower, bark, leaf, seed by conducting MTT assay method along with other pharmacognostical studies. The methanolic extracts of bark, flowers, leaves, and seeds were used for the study. The cell viability MTT assay was performed using the standard operating procedures. The extracts were dissolved in DMSO and serially diluted with complete medium to get the concentrations range of test concentration. DMSO concentration was kept < 0.1% in all the samples. HUVEC cells maintained in appropriate conditions were seeded in 96 well plates and treated with different concentrations of the test samples and incubated at 37°C, 5% CO₂ for 96 hours. MTT reagent was added to the wells and incubated for 4 hours; the dark blue formazan product formed by the cells was dissolved in DMSO under a safety cabinet and read at 550nm. Percentage inhibitions were calculated and plotted with the concentrations used to calculate the IC50 values. The bark, flower, leaves and seed extracts have shown the cytotoxicity activity and can be further studied for antiangiogenesis activity.

Keywords: pharmacognosy, Cell viability, MTT assay, anti-angiogenesis

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2364 Inhibitory Effects of Crocin from Crocus sativus L. on Cell Proliferation of a Medulloblastoma Human Cell Line

Authors: Kyriaki Hatziagapiou, Eleni Kakouri, Konstantinos Bethanis, Alexandra Nikola, Eleni Koniari, Charalabos Kanakis, Elias Christoforides, George Lambrou, Petros Tarantilis

Abstract:

Medulloblastoma is a highly invasive tumour, as it tends to disseminate throughout the central nervous system early in its course. Despite the high 5-year-survival rate, a significant number of patients demonstrate serious long- or short-term sequelae (e.g., myelosuppression, endocrine dysfunction, cardiotoxicity, neurological deficits and cognitive impairment) and higher mortality rates, unrelated to the initial malignancy itself but rather to the aggressive treatment. A strong rationale exists for the use of Crocus sativus L (saffron) and its bioactive constituents (crocin, crocetin, safranal) as pharmaceutical agents, as they exert significant health-promoting properties. Crocins are water soluble carotenoids. Unlike other carotenoids, crocins are highly water-soluble compounds, with relatively low toxicity as they are not stored in adipose and liver tissues. Crocins have attracted wide attention as promising anti-cancer agents, due to their antioxidant, anti-inflammatory, and immunomodulatory effects, interference with transduction pathways implicated in tumorigenesis, angiogenesis, and metastasis (disruption of mitotic spindle assembly, inhibition of DNA topoisomerases, cell-cycle arrest, apoptosis or cell differentiation) and sensitization of cancer cells to radiotherapy and chemotherapy. The current research aimed to study the potential cytotoxic effect of crocins on TE671 medulloblastoma cell line, which may be useful in the optimization of existing and development of new therapeutic strategies. Crocins were extracted from stigmas of saffron in ultrasonic bath, using petroleum-ether, diethylether and methanol 70%v/v as solvents and the final extract was lyophilized. Identification of crocins according to high-performance liquid chromatography (HPLC) analysis was determined comparing the UV-vis spectra and the retention time (tR) of the peaks with literature data. For the biological assays crocin was diluted to nuclease and protease free water. TE671 cells were incubated with a range of concentrations of crocins (16, 8, 4, 2, 1, 0.5 and 0.25 mg/ml) for 24, 48, 72 and 96 hours. Analysis of cell viability after incubation with crocins was performed with Alamar Blue viability assay. The active ingredient of Alamar Blue, resazurin, is a blue, nontoxic, cell permeable compound virtually nonfluorescent. Upon entering cells, resazurin is reduced to a pink and fluorescent molecule, resorufin. Viable cells continuously convert resazurin to resorufin, generating a quantitative measure of viability. The colour of resorufin was quantified by measuring the absorbance of the solution at 600 nm with a spectrophotometer. HPLC analysis indicated that the most abundant crocins in our extract were trans-crocin-4 and trans-crocin-3. Crocins exerted significant cytotoxicity in a dose and time-dependent manner (p < 0.005 for exposed cells to any concentration at 48, 72 and 96 hours versus cells not exposed); as their concentration and time of exposure increased, the reduction of resazurin to resofurin decreased, indicating reduction in cell viability. IC50 values for each time point were calculated ~3.738, 1.725, 0.878 and 0.7566 mg/ml at 24, 48, 72 and 96 hours, respectively. The results of our study could afford the basis of research regarding the use of natural carotenoids as anticancer agents and the shift to targeted therapy with higher efficacy and limited toxicity. Acknowledgements: The research was funded by Fellowships of Excellence for Postgraduate Studies IKY-Siemens Programme.

Keywords: crocetin, crocin, medulloblastoma, saffron

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2363 Mesoporous BiVO4 Thin Films as Efficient Visible Light Driven Photocatalyst

Authors: Karolina Ordon, Sandrine Coste, Malgorzata Makowska-Janusik, Abdelhadi Kassiba

Abstract:

Photocatalytic processes play key role in the production of a new source of energy (as hydrogen), design of self-cleaning surfaces or for the environment preservation. The most challenging task deals with the purification of water distinguished by high efficiency. In the mentioned process, organic pollutants in solutions are decomposed to the simple, non-toxic compounds as H2O and CO2. The most known photocatalytic materials are ZnO, CdS and TiO2 semiconductors with a particular involvement of TiO2 as an efficient photocatalysts even with a high band gap equal to 3.2 eV which exploit only UV radiation from solar emitted spectrum. However, promising material with visible light induced photoactivity was searched through the monoclinic polytype of BiVO4 which has energy gap about 2.4 eV. As required in heterogeneous photocatalysis, the high contact surface is required. Also, BiVO4 as photocatalyst can be optimized by increasing its surface area by achieving the mesoporous structure synthesize. The main goal of the present work consists in the synthesis and characterization of BiVO4 mesoporous thin film. The synthesis method based on sol-gel was carried out using a standard surfactants such as P123 and F127. The thin film was deposited by spin and dip coating method. Then, the structural analysis of the obtained material was performed thanks to X-ray diffraction (XRD) and Raman spectroscopy. The surface of resulting structure was investigated using a scanning electron microscopy (SEM). The computer simulations based on modeling the optical and electronic properties of bulk BiVO4 by using DFT (density functional theory) methodology were carried out. The semiempirical parameterized method PM6 was used to compute the physical properties of BiVO4 nanostructures. The Raman and IR absorption spectra were also measured for synthesized mesoporous material, and the results were compared with the theoretical predictions. The simulations of nanostructured BiVO4 have pointed out the occurrence of quantum confinement for nanosized clusters leading to widening of the band gap. This result overcame the relevance of nanosized objects to harvest wide part of the solar spectrum. Also, a balance was searched experimentally through the mesoporous nature of the films devoted to enhancing the contact surface as required for heterogeneous catalysis without to lower the nanocrystallite size under some critical sizes inducing an increased band gap. The present contribution will discuss the relevant features of the mesoporous films with respect to their photocatalytic responses.

Keywords: bismuth vanadate, photocatalysis, thin film, quantum-chemical calculations

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2362 Evaluation of Suspended Particles Impact on Condensation in Expanding Flow with Aerodynamics Waves

Authors: Piotr Wisniewski, Sławomir Dykas

Abstract:

Condensation has a negative impact on turbomachinery efficiency in many energy processes.In technical applications, it is often impossible to dry the working fluid at the nozzle inlet. One of the most popular working fluid is atmospheric air that always contains water in form of steam, liquid, or ice crystals. Moreover, it always contains some amount of suspended particles which influence the phase change process. It is known that the phenomena of evaporation or condensation are connected with release or absorption of latent heat, what influence the fluid physical properties and might affect the machinery efficiency therefore, the phase transition has to be taken under account. This researchpresents an attempt to evaluate the impact of solid and liquid particles suspended in the air on the expansion of moist air in a low expansion rate, i.e., with expansion rate, P≈1000s⁻¹. The numerical study supported by analytical and experimental research is presented in this work. The experimental study was carried out using an in-house experimental test rig, where nozzle was examined for different inlet air relative humidity values included in the range of 25 to 51%. The nozzle was tested for a supersonic flow as well as for flow with shock waves induced by elevated back pressure. The Schlieren photography technique and measurement of static pressure on the nozzle wall were used for qualitative identification of both condensation and shock waves. A numerical model validated against experimental data available in the literature was used for analysis of occurring flow phenomena. The analysis of the suspended particles number, diameter, and character (solid or liquid) revealed their connection with heterogeneous condensation importance. If the expansion of fluid without suspended particlesis considered, the condensation triggers so called condensation wave that appears downstream the nozzle throat. If the solid particles are considered, with increasing number of them, the condensation triggers upwind the nozzle throat, decreasing the condensation wave strength. Due to the release of latent heat during condensation, the fluid temperature and pressure increase, leading to the shift of normal shock upstream the flow. Owing relatively large diameters of the droplets created during heterogeneous condensation, they evaporate partially on the shock and continues to evaporate downstream the nozzle. If the liquid water particles are considered, due to their larger radius, their do not affect the expanding flow significantly, however might be in major importance while considering the compression phenomena as they will tend to evaporate on the shock wave. This research proves the need of further study of phase change phenomena in supersonic flow especially considering the interaction of droplets with the aerodynamic waves in the flow.

Keywords: aerodynamics, computational fluid dynamics, condensation, moist air, multi-phase flows

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2361 Rapid, Label-Free, Direct Detection and Quantification of Escherichia coli Bacteria Using Nonlinear Acoustic Aptasensor

Authors: Shilpa Khobragade, Carlos Da Silva Granja, Niklas Sandström, Igor Efimov, Victor P. Ostanin, Wouter van der Wijngaart, David Klenerman, Sourav K. Ghosh

Abstract:

Rapid, label-free and direct detection of pathogenic bacteria is critical for the prevention of disease outbreaks. This paper for the first time attempts to probe the nonlinear acoustic response of quartz crystal resonator (QCR) functionalized with specific DNA aptamers for direct detection and quantification of viable E. coli KCTC 2571 bacteria. DNA aptamers were immobilized through biotin and streptavidin conjugation, onto the gold surface of QCR to capture the target bacteria and the detection was accomplished by shift in amplitude of the peak 3f signal (3 times the drive frequency) upon binding, when driven near fundamental resonance frequency. The developed nonlinear acoustic aptasensor system demonstrated better reliability than conventional resonance frequency shift and energy dissipation monitoring that were recorded simultaneously. This sensing system could directly detect 10⁽⁵⁾ cells/mL target bacteria within 30 min or less and had high specificity towards E. coli KCTC 2571 bacteria as compared to the same concentration of S.typhi bacteria. Aptasensor response was observed for the bacterial suspensions ranging from 10⁽⁵⁾-10⁽⁸⁾ cells/mL. Conclusively, this nonlinear acoustic aptasensor is simple to use, gives real-time output, cost-effective and has the potential for rapid, specific, label-free direction detection of bacteria.

Keywords: acoustic, aptasensor, detection, nonlinear

Procedia PDF Downloads 556