Search results for: rat red blood cell haemoglobin
Commenced in January 2007
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Edition: International
Paper Count: 5611

Search results for: rat red blood cell haemoglobin

4771 Nafion Nanofiber Composite Membrane Fabrication for Fuel Cell Applications

Authors: C. N. Okafor, M. Maaza, T. A. E. Mokrani

Abstract:

A proton exchange membrane has been developed for Direct Methanol Fuel Cell (DMFC). The nanofiber network composite membranes were prepared by interconnected network of Nafion (perfuorosulfonic acid) nanofibers that have been embedded in an uncharged and inert polymer matrix, by electro-spinning. The spinning solution of Nafion with a low concentration (1 wt. % compared to Nafion) of high molecular weight poly(ethylene oxide), as a carrier polymer. The interconnected network of Nafion nanofibers with average fiber diameter in the range of 160-700nm, were used to make the membranes, with the nanofiber occupying up to 85% of the membrane volume. The matrix polymer was cross-linked with Norland Optical Adhesive 63 under UV. The resulting membranes showed proton conductivity of 0.10 S/cm at 25°C and 80% RH; and methanol permeability of 3.6 x 10-6 cm2/s.

Keywords: composite membrane, electrospinning, fuel cell, nanofibers

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4770 Shear Stress and Effective Structural Stress ‎Fields of an Atherosclerotic Coronary Artery

Authors: Alireza Gholipour, Mergen H. Ghayesh, Anthony Zander, Stephen J. Nicholls, Peter J. Psaltis

Abstract:

A three-dimensional numerical model of an atherosclerotic coronary ‎artery is developed for the determination of high-risk situation and ‎hence heart attack prediction. Employing the finite element method ‎‎(FEM) using ANSYS, fluid-structure interaction (FSI) model of the ‎artery is constructed to determine the shear stress distribution as well ‎as the von Mises stress field. A flexible model for an atherosclerotic ‎coronary artery conveying pulsatile blood is developed incorporating ‎three-dimensionality, artery’s tapered shape via a linear function for ‎artery wall distribution, motion of the artery, blood viscosity via the ‎non-Newtonian flow theory, blood pulsation via use of one-period ‎heartbeat, hyperelasticity via the Mooney-Rivlin model, viscoelasticity ‎via the Prony series shear relaxation scheme, and micro-calcification ‎inside the plaque. The material properties used to relate the stress field ‎to the strain field have been extracted from clinical data from previous ‎in-vitro studies. The determined stress fields has potential to be used as ‎a predictive tool for plaque rupture and dissection.‎ The results show that stress concentration due to micro-calcification ‎increases the von Mises stress significantly; chance of developing a ‎crack inside the plaque increases. Moreover, the blood pulsation varies ‎the stress distribution substantially for some cases.‎

Keywords: atherosclerosis, fluid-structure interaction‎, coronary arteries‎, pulsatile flow

Procedia PDF Downloads 153
4769 Transcriptional Differences in B cell Subpopulations over the Course of Preclinical Autoimmunity Development

Authors: Aleksandra Bylinska, Samantha Slight-Webb, Kevin Thomas, Miles Smith, Susan Macwana, Nicolas Dominguez, Eliza Chakravarty, Joan T. Merrill, Judith A. James, Joel M. Guthridge

Abstract:

Background: Systemic Lupus Erythematosus (SLE) is an interferon-related autoimmune disease characterized by B cell dysfunction. One of the main hallmarks is a loss of tolerance to self-antigens leading to increased levels of autoantibodies against nuclear components (ANAs). However, up to 20% of healthy ANA+ individuals will not develop clinical illness. SLE is more prevalent among women and minority populations (African, Asian American and Hispanics). Moreover, African Americans have a stronger interferon (IFN) signature and develop more severe symptoms. The exact mechanisms involved in ethnicity-dependent B cell dysregulation and the progression of autoimmune disease from ANA+ healthy individuals to clinical disease remains unclear. Methods: Peripheral blood mononuclear cells (PBMCs) from African (AA) and European American (EA) ANA- (n=12), ANA+ (n=12) and SLE (n=12) individuals were assessed by multimodal scRNA-Seq/CITE-Seq methods to examine differential gene signatures in specific B cell subsets. Library preparation was done with a 10X Genomics Chromium according to established protocols and sequenced on Illumina NextSeq. The data were further analyzed for distinct cluster identification and differential gene signatures in the Seurat package in R and pathways analysis was performed using Ingenuity Pathways Analysis (IPA). Results: Comparing all subjects, 14 distinct B cell clusters were identified using a community detection algorithm and visualized with Uniform Manifold Approximation Projection (UMAP). The proportion of each of those clusters varied by disease status and ethnicity. Transitional B cells trended higher in ANA+ healthy individuals, especially in AA. Ribonucleoprotein high population (HNRNPH1 elevated, heterogeneous nuclear ribonucleoprotein, RNP-Hi) of proliferating Naïve B cells were more prevalent in SLE patients, specifically in EA. Interferon-induced protein high population (IFIT-Hi) of Naive B cells are increased in EA ANA- individuals. The proportion of memory B cells and plasma cells clusters tend to be expanded in SLE patients. As anticipated, we observed a higher signature of cytokine-related pathways, especially interferon, in SLE individuals. Pathway analysis among AA individuals revealed an NRF2-mediated Oxidative Stress response signature in the transitional B cell cluster, not seen in EA individuals. TNFR1/2 and Sirtuin Signaling pathway genes were higher in AA IFIT-Hi Naive B cells, whereas they were not detected in EA individuals. Interferon signaling was observed in B cells in both ethnicities. Oxidative phosphorylation was found in age-related B cells (ABCs) for both ethnicities, whereas Death Receptor Signaling was found only in EA patients in these cells. Interferon-related transcription factors were elevated in ABCs and IFIT-Hi Naive B cells in SLE subjects of both ethnicities. Conclusions: ANA+ healthy individuals have altered gene expression pathways in B cells that might drive apoptosis and subsequent clinical autoimmune pathogenesis. Increases in certain regulatory pathways may delay progression to SLE. Further, AA individuals have more elevated activation pathways that may make them more susceptible to SLE.

Keywords:

Procedia PDF Downloads 157
4768 The Role of Autophagy Modulation in Angiotensin-II Induced Hypertrophy

Authors: Kitti Szoke, Laszlo Szoke, Attila Czompa, Arpad Tosaki, Istvan Lekli

Abstract:

Autophagy plays an important role in cardiac hypertrophy, which is one of the most common causes of heart failure in the world. This self-degradative catabolic process, responsible for protein quality control, balancing sources of energy at critical times, and elimination of damaged organelles. The autophagic activity can be triggered by starvation, oxidative stress, or pharmacological agents, like rapamycin. This induced autophagy can promote cell survival during starvation or pathological stress. In this study, it is investigated the effect of the induced autophagic process on angiotensin induced hypertrophic H9c2 cells. In our study, it is used H9c2 cells as an in vitro model. To induce hypertrophy, cells were treated with 10000 nM angiotensin-II, and to activate autophagy, 100 nM rapamycin treatment was used. The following groups were formed: 1: control, 2: 10000 nM AT-II, 3: 100 nM rapamycin, 4: 100 nM rapamycin pretreatment then 10000 nM AT-II. The cell viability was examined via MTT (cell proliferation assay) assay. The cells were stained with rhodamine-conjugated phalloidin and DAPI to visualize F-actin filaments and cell nuclei then the cell size alteration was examined in a fluorescence microscope. Furthermore, the expression levels of autophagic and apoptotic proteins such as Beclin-1, p62, LC3B-II, Cleaved Caspase-3 were evaluated by Western blot. MTT assay result suggests that the used pharmaceutical agents in the tested concentrations did not have a toxic effect; however, at group 3, a slight decrement was detected in cell viability. In response to AT-II treatment, a significant increase was detected in the cell size; cells became hypertrophic. However, rapamycin pretreatment slightly reduced the cell size compared to group 2. Western blot results showed that AT-II treatment-induced autophagy, because the increased expression of Beclin-1, p62, LC3B-II were observed. However, due to the incomplete autophagy, the apoptotic Cleaved Caspase-3 expression also increased. Rapamycin pretreatment up-regulated Beclin-1 and LC3B-II, down-regulated p62 and Cleaved Caspase-3, indicating that rapamycin-induced autophagy can restore the normal autophagic flux. Taken together, our results suggest that rapamycin activated autophagy reduces angiotensin-II induced hypertrophy.

Keywords: angiotensin-II, autophagy, H9c2 cell line, hypertrophy, rapamycin

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4767 Recent Development of Materials for Proton Exchange Membrane Fuel Cell (PEMFC)

Authors: Mohammed Jourdani, Hamid Mounir, Abdellatif El Marjani

Abstract:

Proton exchange membrane fuel cells (PEMFCs) have been developed as a promising power source for transportation and stationary applications, and power devices for computers and mobile telephones. This paper discusses and summarizes the latest developments of materials and remaining challenges of PEMFC. The different contributions to the material of all components and the efficiencies are analyzed. Many technical advances are introduced to increase the PEMFC fuel cell efficiency and life time for transportation, stationary and portable utilization. By the last years the total cost of this system is decreasing. However, the remaining challenges that need to be overcome mean that it will be several years before full commercialization can take place.

Keywords: PEMFC fuel cell, materials, recent development, efficiency, life time, commercialization possibility

Procedia PDF Downloads 290
4766 The Effect of Antibiotic Use on Blood Cultures: Implications for Future Policy

Authors: Avirup Chowdhury, Angus K. McFadyen, Linsey Batchelor

Abstract:

Blood cultures (BCs) are an important aspect of management of the septic patient, identifying the underlying pathogen and its antibiotic sensitivities. However, while the current literature outlines indications for initial BCs to be taken, there is little guidance for repeat sampling in the following 5-day period and little information on how antibiotic use can affect the usefulness of this investigation. A retrospective cohort study was conducted using inpatients who had undergone 2 or more BCs within 5 days between April 2016 and April 2017 at a 400-bed hospital in the west of Scotland and received antibiotic therapy between the first and second BCs. The data for BC sampling was collected from the electronic microbiology database, and cross-referenced with data from the hospital electronic prescribing system. Overall, 283 BCs were included in the study, taken from 92 patients (mean 3.08 cultures per patient, range 2-10). All 92 patients had initial BCs, of which 83 were positive (90%). 65 had a further sample within 24 hours of commencement of antibiotics, with 35 positive (54%). 23 had samples within 24-48 hours, with 4 (17%) positive; 12 patients had sampling at 48-72 hours, 12 at 72-96 hours, and 10 at 96-120 hours, with none positive. McNemar’s Exact Test was used to calculate statistical significance for patients who received blood cultures in multiple time blocks (Initial, < 24h, 24-120h, > 120h). For initial vs. < 24h-post BCs (53 patients tested), the proportion of positives fell from 46/53 to 29/53 (one-tailed P=0.002, OR 3.43, 95% CI 1.48-7.96). For initial vs 24-120h (n=42), the proportions were 38/42 and 4/42 respectively (P < 0.001, OR 35.0, 95% CI 4.79-255.48). For initial vs > 120h (n=36), these were 33/36 and 2/36 (P < 0.001,OR ∞). These were also calculated for a positive in initial or < 24h vs. 24-120h (n=42), with proportions of 41/42 and 4/42 (P < 0.001, OR 38.0, 95% CI 5.22-276.78); and for initial or < 24h vs > 120h (n=36), with proportions of 35/36 and 2/36 respectively (P < 0.001, OR ∞). This data appears to show that taking an initial BC followed by a BC within 24 hours of antibiotic commencement would maximise blood culture yield while minimising the risk of false negative results. This could potentially remove the need for as many as 46% of BC samples without adversely affecting patient care. BC yield decreases sharply after 48 hours of antibiotic use, and may not provide any clinically useful information after this time. Further multi-centre studies would validate these findings, and provide a foundation for future health policy generation.

Keywords: antibiotics, blood culture, efficacy, inpatient

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4765 Phase Equilibria in the Ln-Sr-Co-O Systems

Authors: Anastasiia Maklakova

Abstract:

The perovskite type oxides formed in the Ln-Me-Me/-O systems (where Ln – rare-earth, Me – alkaline earth metal, Me/ - 3-d metal) have potential applications as gas sensors, catalysts or cathode materials for IT-SOFCs due to the high values of mixed electronic -ionic conductivity and high oxygen diffusivity. Complex oxides in the Sr-(Pr,Gd)-Co-O systems were prepared via the glycerol-nitrate technique The phase composition was determined using a Shimadzu XRD-7000 diffractometer at room temperature in air. Phase identification was performed using the ICDD database. The structure was refined by the full-profile Rietveld method using Fullprof 2008 software. Gradual substitution of strontium by Pr or Gd leads to the decrease of unit cell parameters and unit cell volume that can be explained by the size factor. An introduction of Pr or Gd into the strontium cobaltite increases the oxygen content in samples.

Keywords: phase equilibria, crystal structure, oxygen nonstoichiometry, solid oxide fuel cell

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4764 Effects of Cuminum cyminum L. Essential Oil Supplementation on Components of Metabolic Syndrome: A Clinical Trial

Authors: Ashti Morovati, Hushyar Azari, Bahram Pourghassem Gargari

Abstract:

Objectives and goals: The prevalence of metabolic syndrome (MetS), as a major health burden for societies, is increasing. This clinical trial was conducted to evaluate the effects of CuEO supplementation on anthropometric indices, systolic and diastolic blood pressure, blood glucose level, insulin resistance and serum lipid level in patients suffering from MetS. Methods: This was a randomized, triple‐blind, placebo‐controlled clinical trial in which 56 patients with MetS aged 18–60 years who fulfilled the eligibility criteria were randomly allocated to an intervention or a control group. Inclusion criteria for the study were comprised of diagnosis of MetS according to the new International Federation of Diabetes. The exclusion criteria were defined as: taking herbal supplements, use of drugs having evident interaction with cumin such as anti‐depressant drugs, vitamin D, omega 3, selenium, zinc, smoking, pregnancy, or breastfeeding, suffering from cancer, having any history of gastrointestinal and hepatic, cardiovascular, thyroid and kidney disorders, and menopause. 75 mg CuEO or placebo soft gels were administered three times daily to the participants for eight weeks. The soft gel consumption was checked by asking the participants to bring the medication containers in the follow‐up visits at the 4th and the 8th weeks of the study. Data pertaining to blood pressure, height, weight, waist circumference, hip circumference and BMI, as well as food consumption were collected at the beginning and end of the study. Fasting blood samples ( glucose, triglyceride, total cholesterol, HDL-cholesterol and LDL-cholesterol) were obtained and biochemical measurements were assessed at the beginning and end of the study. Results: At eight weeks, a total of 44 patients completed this study. Except for diastolic blood pressure (DBP), the other assessed variables were not significantly different between the two groups. In intra group analysis, placebo and CuEO groups both had insignificant decrements in DBP (mean difference [MD] with 95% CI: −3.31 [−7.11, 0.47] and −1.77 [−5.95, 2.40] mmHg, respectively). However, DBP was significantly lower in CuEO compared with the placebo group at the end of study (81.41 ± 5.88 vs. 84.09 ± 5.54 mmHg, MD with 95% CI: −3.98 [−7.60, −0.35] mmHg, p < .05). Conclusions: The results of this study indicated that CuEO does not have any effect on MetS components, except for DBP in patients with MetS.

Keywords: blood pressure, fasting blood glucose, lipid profile, waist circumference

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4763 Deciphering Tumor Stroma Interactions in Retinoblastoma

Authors: Rajeswari Raguraman, Sowmya Parameswaran, Krishnakumar Subramanian, Jagat Kanwar, Rupinder Kanwar

Abstract:

Background: Tumor microenvironment has been implicated in several cancers to regulate cell growth, invasion and metastasis culminating in outcome of therapy. Tumor stroma consists of multiple cell types that are in constant cross-talk with the tumor cells to favour a pro-tumorigenic environment. Not much is known about the existence of tumor microenvironment in the pediatric intraocular malignancy, Retinoblastoma (RB). In the present study, we aim to understand the multiple stromal cellular subtypes and tumor stromal interactions expressed in RB tumors. Materials and Methods: Immunohistochemistry for stromal cell markers CD31, CD68, alpha-smooth muscle (α-SMA), vimentin and glial fibrillary acidic protein (GFAP) was performed on formalin fixed paraffin embedded tissues sections of RB (n=12). The differential expression of stromal target molecules; fibroblast activation protein (FAP), tenascin-C (TNC), osteopontin (SPP1), bone marrow stromal antigen 2 (BST2), stromal derived factor 2 and 4 (SDF2 and SDF4) in primary RB tumors (n=20) and normal retina (n=5) was studied by quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) and Western blotting. The differential expression was correlated with the histopathological features of RB. The interaction between RB cell lines (Weri-Rb-1, NCC-RbC-51) and Bone marrow stromal cells (BMSC) was also studied using direct co-culture and indirect co-culture methods. The functional effect of the co-culture methods on the RB cells was evaluated by invasion and proliferation assays. Global gene expression was studied by using Affymetrix 3’ IVT microarray. Pathway prediction was performed using KEGG and the key molecules were validated using qRT-PCR. Results: The immunohistochemistry revealed the presence of several stromal cell types such as endothelial cells (CD31+;Vim+/-); macrophages (CD68+;Vim+/-); Fibroblasts (Vim+; CD31-;CD68- );myofibroblasts (α-SMA+/ Vim+) and invading retinal astrocytes/ differentiated retinal glia (GFAP+; Vim+). A characteristic distribution of these stromal cell types was observed in the tumor microenvironment, with endothelial cells predominantly seen in blood vessels and macrophages near actively proliferating tumor or necrotic areas. Retinal astrocytes and glia were predominant near the optic nerve regions in invasive tumors with sparse distribution in tumor foci. Fibroblasts were widely distributed with rare evidence of myofibroblasts in the tumor. Both gene and protein expression revealed statistically significant (P<0.05) up-regulation of FAP, TNC and BST2 in primary RB tumors compared to the normal retina. Co-culture of BMSC with RB cells promoted invasion and proliferation of RB cells in direct and indirect contact methods respectively. Direct co-culture of RB cell lines with BMSC resulted in gene expression changes in ECM-receptor interaction, focal adhesion, IL-8 and TGF-β signaling pathways associated with cancer. In contrast, various metabolic pathways such a glucose, fructose and amino acid metabolism were significantly altered under the indirect co-culture condition. Conclusion: The study suggests that the close interaction between RB cells and the stroma might be involved in RB tumor invasion and progression which is likely to be mediated by ECM-receptor interactions and secretory factors. Targeting the tumor stroma would be an attractive option for redesigning treatment strategies for RB.

Keywords: gene expression profiles, retinoblastoma, stromal cells, tumor microenvironment

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4762 Cytotoxicity of Flavonoid Compounds from Smilax corbularia Kunth Against Cholangiocarcinoma Cell Line

Authors: Pakakrong Thongdeeying, Srisopa Ruangnoo, Arunporn Itharat

Abstract:

The rhizomes of Smilax corbularia Kunth have long been used as common ingredients in anticancer preparations. Thus, the objective of this study is to investigate cytotoxicity of S. corbularia and its ingredients against cholangiocarcinoma cell line (KKU-M156) by SRB assay. Ethanolic and water extracts of S. corbularia rhizomes were obtained using the procedures followed by Thai traditional doctors. Bioassay guided isolation was used to isolate cytotoxic compounds. The results revealed that the ethanolic extract of S. corbularia exhibited activity against KKU-M156 cell line with an IC50 value of 84.53±1.62 µg/ml, but the water extract showed no cytotoxic activity. Three flavonoid compounds [astilbin (1), engeletin (2), and quercetin (3)] were isolated from the ethanolic extract. Compound 3 exhibited the strongest activity against KKU-M156 cell line (IC50 = 8.14 ± 1.15 µg/ml), but 1 and 2 showed no cytotoxic activity (IC50 > 100 µg/ml). In conclusion, quercetin showed the highest efficacy against cholangiocarcinoma. These results support the traditional use of this plant by Thai traditional doctors for cancer treatment.

Keywords: cholangiocarcinoma, cytotoxicity, flavonoid, Smilax corbularia

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4761 Robust Diagnosability of PEMFC Based on Bond Graph LFT

Authors: Ould Bouamama, M. Bressel, D. Hissel, M. Hilairet

Abstract:

Fuel cell (FC) is one of the best alternatives of fossil energy. Recently, the research community of fuel cell has shown a considerable interest for diagnosis in view to ensure safety, security, and availability when faults occur in the process. The problematic for model based FC diagnosis consists in that the model is complex because of coupling of several kind of energies and the numerical values of parameters are not always known or are uncertain. The present paper deals with use of one tool: the Linear Fractional Transformation bond graph tool not only for uncertain modelling but also for monitorability (ability to detect and isolate faults) analysis and formal generation of robust fault indicators with respect to parameter uncertainties.The developed theory applied to a nonlinear FC system has proved its efficiency.

Keywords: bond graph, fuel cell, fault detection and isolation (FDI), robust diagnosis, structural analysis

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4760 Microfluidic Based High Throughput Screening System for Photodynamic Therapy against Cancer Cells

Authors: Rina Lee, Chung-Hun Oh, Eunjin Lee, Jeongyun Kim

Abstract:

The Photodynamic therapy (PDT) is a treatment that uses a photosensitizer as a drug to damage and kill cancer cells. After injecting the photosensitizer into the bloodstream, the drug is absorbed by cancer cells selectively. Then the area to be treated is exposed to specific wavelengths of light and the photosensitizer produces a form of oxygen that kills nearby cancer cells. PDT is has an advantage to destroy the tumor with minimized side-effects on normal cells. But, PDT is not a completed method for cancer therapy. Because the mechanism of PDT is quite clear yet and the parameters such as intensity of light and dose of photosensitizer are not optimized for different types of cancers. To optimize these parameters, we suggest a novel microfluidic system to automatically control intensity of light exposure with a personal computer (PC). A polydimethylsiloxane (PDMS) microfluidic chip is composed with (1) a cell culture channels layer where cancer cells were trapped to be tested with various dosed photofrin (1μg/ml used for the test) as the photosensitizer and (2) a color dye layer as a neutral density (ND) filter to reduce intensity of light which exposes the cell culture channels filled with cancer cells. Eight different intensity of light (10%, 20%, …, 100%) are generated through various concentrations of blue dye filling the ND filter. As a light source, a light emitting diode (LED) with 635nm wavelength was placed above the developed PDMS microfluidic chip. The total time for light exposure was 30 minutes and HeLa and PC3 cell lines of cancer cells were tested. The cell viability of cells was evaluated with a Live/Dead assay kit (L-3224, Invitrogen, USA). The stronger intensity of light exposed, the lower viability of the cell was observed, and vice versa. Therefore, this system was demonstrated through investigating the PDT against cancer cell to optimize the parameters as critical light intensity and dose of photosensitizer. Our results suggest that the system can be used for optimizing the combinational parameters of light intensity and photosensitizer dose against diverse cancer cell types.

Keywords: photodynamic therapy, photofrin, high throughput screening, hela

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4759 Direct Phoenix Identification and Antimicrobial Susceptibility Testing from Positive Blood Culture Broths

Authors: Waad Al Saleemi, Badriya Al Adawi, Zaaima Al Jabri, Sahim Al Ghafri, Jalila Al Hadhramia

Abstract:

Objectives: Using standard lab methods, a positive blood culture requires a minimum of two days (two occasions of overnight incubation) to obtain a final identification (ID) and antimicrobial susceptibility results (AST) report. In this study, we aimed to evaluate the accuracy and precision of identification and antimicrobial susceptibility testing of an alternative method (direct method) that will reduce the turnaround time by 24 hours. This method involves the direct inoculation of positive blood culture broths into the Phoenix system using serum separation tubes (SST). Method: This prospective study included monomicrobial-positive blood cultures obtained from January 2022 to May 2023 in SQUH. Blood cultures containing a mixture of organisms, fungi, or anaerobic organisms were excluded from this study. The result of the new “direct method” under study was compared with the current “standard method” used in the lab. The accuracy and precision were evaluated for the ID and AST using Clinical and Laboratory Standards Institute (CLSI) recommendations. The categorical agreement, essential agreement, and the rates of very major errors (VME), major errors (ME), and minor errors (MIE) for both gram-negative and gram-positive bacteria were calculated. Passing criteria were set according to CLSI. Result: The results of ID and AST were available for a total of 158 isolates. Of 77 isolates of gram-negative bacteria, 71 (92%) were correctly identified at the species level. Of 70 isolates of gram-positive bacteria, 47(67%) isolates were correctly identified. For gram-negative bacteria, the essential agreement of the direct method was ≥92% when compared to the standard method, while the categorical agreement was ≥91% for all tested antibiotics. The precision of ID and AST were noted to be 100% for all tested isolates. For gram-positive bacteria, the essential agreement was >93%, while the categorical agreement was >92% for all tested antibiotics except moxifloxacin. Many antibiotics were noted to have an unacceptable higher rate of very major errors including penicillin, cotrimoxazole, clindamycin, ciprofloxacin, and moxifloxacin. However, no error was observed in the results of vancomycin, linezolid, and daptomycin. Conclusion: The direct method of ID and AST for positive blood cultures using SST is reliable for gram negative bacteria. It will significantly decrease the turnaround time and will facilitate antimicrobial stewardship.

Keywords: bloodstream infection, oman, direct ast, blood culture, rapid identification, antimicrobial susceptibility, phoenix, direct inoculation

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4758 Differential Diagnosis of Malaria and Dengue Fever on the Basis of Clinical Findings and Laboratory Investigations

Authors: Aman Ullah Khan, Muhammad Younus, Aqil Ijaz, Muti-Ur-Rehman Khan, Sayyed Aun Muhammad, Asif Idrees, Sanan Raza, Amar Nasir

Abstract:

Dengue fever and malaria are important vector-borne diseases of public health significance affecting millions of people around the globe. Dengue fever is caused by Dengue virus while malaria is caused by plasmodium protozoan. Generally, the consequences of Malaria are less severe compared to dengue fever. This study was designed to differentiate dengue fever and malaria on the basis of clinical and laboratory findings and to compare the changes in both diseases having different causative agents transmitted by the common vector. A total of 200 patients of dengue viral infection (120 males, 80 females) were included in this prospective descriptive study. The blood samples of the individuals were first screened for malaria by blood smear examination and then the negative samples were tested by anti-dengue IgM strip. The strip positive cases were further screened by IgM capture ELISA and their complete blood count including hemoglobin estimation (Hb), total and differential leukocyte counts (TLC and DLC), erythrocyte sedimentation rate (ESR) and platelet counts were performed. On the basis of the severity of signs and symptoms, dengue virus infected patients were subdivided into dengue fever (DF) and dengue hemorrhagic fever (DHF) comprising 70 and 100 confirmed patients, respectively. On the other hand, 30 patients were found infected with Malaria while overall 120 patients showed thrombocytopenia. The patients of DHF were found to have more leucopenia, raised hemoglobin level and thrombocytopenia < 50,000/µl compared to the patients belonging to DF and malaria. On the basis of the outcomes of the study, it was concluded that patients affected by DF were at a lower risk of undergoing haematological disturbance than suffering from DHF. While, the patients infected by Malaria were found to have no significant change in their blood components.

Keywords: dengue fever, blood, serum, malaria, ELISA

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4757 Role of Micro-Patterning on Stem Cell-Material Interaction Modulation and Cell Fate

Authors: Lay Poh Tan, Chor Yong Tay, Haiyang Yu

Abstract:

Micro-contact printing is a form of soft lithography that uses the relief patterns on a master polydimethylsiloxane (PDMS) stamp to form patterns of self-assembled monolayers (SAMs) of ink on the surface of a substrate through conformal contact technique. Here, we adopt this method to print proteins of different dimensions on our biodegradable polymer substrates. We started off with printing 20-500 μm scale lanes of fibronectin to engineer the shape of bone marrow derived human mesenchymal stem cell (hMSCs). After 8 hours of culture, the hMSCs adopted elongated shapes, and upon analysis of the gene expressions, genes commonly associated with myogenesis (GATA-4, MyoD1, cTnT and β-MHC) and neurogenesis (NeuroD, Nestin, GFAP, and MAP2) were up-regulated but gene expression associated to osteogenesis (ALPL, RUNX2, and SPARC) were either down modulated or remained at the nominal level. This is the first evidence that cellular morphology control via micropatterning could be used to modulate stem cell fate without external biochemical stimuli. We further our studies to modulate the focal adhesion (FA) instead of the macro shape of cells. Micro-contact printed islands of different smaller dimensions were investigated. We successfully regulated the FAs into dense FAs and elongated FAs by micropatterning. Additionally, the combined effects of hard (40.4 kPa), and intermediate (10.6 kPa) PA gel and FAs patterning on hMSCs differentiation were studied. Results showed that FA and matrix compliance plays an important role in hMSCs differentiation, and there is a cross-talk between different physical stimulants and the significance of these stimuli can only be realized if they are combined at the optimum level.

Keywords: micro-contact printing, polymer substrate, cell-material interaction, stem cell differentiation

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4756 Statistical Manufacturing Cell/Process Qualification Sample Size Optimization

Authors: Angad Arora

Abstract:

In production operations/manufacturing, a cell or line is typically a bunch of similar machines (computer numerical control (CNCs), advanced cutting, 3D printing or special purpose machines. For qualifying a typical manufacturing line /cell / new process, Ideally, we need a sample of parts that can be flown through the process and then we make a judgment on the health of the line/cell. However, with huge volumes and mass production scope, such as in the mobile phone industry, for example, the actual cells or lines can go in thousands and to qualify each one of them with statistical confidence means utilizing samples that are very large and eventually add to product /manufacturing cost + huge waste if the parts are not intended to be customer shipped. To solve this, we come up with 2 steps statistical approach. We start with a small sample size and then objectively evaluate whether the process needs additional samples or not. For example, if a process is producing bad parts and we saw those samples early, then there is a high chance that the process will not meet the desired yield and there is no point in keeping adding more samples. We used this hypothesis and came up with 2 steps binomial testing approach. Further, we also prove through results that we can achieve an 18-25% reduction in samples while keeping the same statistical confidence.

Keywords: statistics, data science, manufacturing process qualification, production planning

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4755 The Response of Adaptive Mechanism of Fluorescent Proteins from Coral Species and Target Cell Properties on Signalling Capacity as Biosensor

Authors: Elif Tugce Aksun Tumerkan

Abstract:

Fluorescent proteins (FPs) have become very popular since green fluorescent protein discovered from crystal jellyfish. It is known that Anthozoa species have a wide range of chromophore organisms, and the initial crystal structure for non-fluorescent chromophores obtained from the reef-building coral has been determined. There are also differently coloured pigments in non-bioluminescent Anthozoa zooxanthellate and azooxanthellate which are frequently members of the GFP-like protein family. The development of fluorescent proteins (FPs) and their applications is an outstanding example of basic science leading to practical biotechnological and medical applications. Fluorescent proteins have several applications in science and are used as important indicators in molecular biology and cell-based research. With rising interest in cell biology, FPs have used as biosensor indicators and probes in pharmacology and cell biology. Using fluorescent proteins in genetically encoded metabolite sensors has many advantages than chemical probes for metabolites such as easily introduced into any cell or organism in any sub-cellular localization and giving chance to fixing to fluoresce of different colours or characteristics. There are different factors effects to signalling mechanism when they used as a biosensor. While there are wide ranges of research have been done on the significance and applications of fluorescent proteins, the cell signalling response of FPs and target cell are less well understood. In this study, it was aimed to clarify the response of adaptive mechanisms of coral species such as pH, temperature and symbiotic relationship and target cells properties on the signalling capacity. Corals are a rich natural source of fluorescent proteins that change with environmental conditions such as light, heat stress and injury. Adaptation mechanism of coral species to these types of environmental variations is important factor due to FPs properties have affected by this mechanism. Since fluorescent proteins obtained from nature, their own ecological property like the symbiotic relationship is observed very commonly in coral species and living conditions have the impact on FPs efficiency. Target cell properties also have an effect on signalling and visualization. The dynamicity of detector that used for reading fluorescence and the level of background fluorescence are key parameters for the quality of the fluorescent signal. Among the factors, it can be concluded that coral species adaptive characteristics have the strongest effect on FPs signalling capacity.

Keywords: biosensor, cell biology, environmental conditions, fluorescent protein, sea anemone

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4754 The Effect of Naringenin on the Apoptosis in T47D Cell Line of Breast Cancer

Authors: AliAkbar Hafezi, Jahanbakhsh Asadi, Majid Shahbazi, Alijan Tabarraei, Nader Mansour Samaei, Hamed Sheibak, Roghaye Gharaei

Abstract:

Background: Breast cancer is the most common cancer in women. In most cancer cells, apoptosis is blocked. As for the importance of apoptosis in cancer cell death and the role of different genes in its induction or inhibition, the search for compounds that can begin the process of apoptosis in tumor cells is discussed as a new strategy in anticancer drug discovery. The aim of this study was to investigate the effect of Naringenin (NGEN) on the apoptosis in the T47D cell line of breast cancer. Materials and Methods: In this experimental study in vitro, the T47D cell line of breast cancer was selected as a sample. The cells at 24, 48, and 72 hours were treated with doses of 20, 200, and 1000 µm of Naringenin. Then, the transcription levels of the genes involved in apoptosis, including Bcl-2, Bax, Caspase 3, Caspase 8, Caspase 9, P53, PARP-1, and FAS, were assessed using Real Time-PCR. The collected data were analyzed using IBM SPSS Statistics 24.0. Results: The results showed that Naringenin at doses of 20, 200, and 1000 µm in all three times of 24, 48, and 72 hours increased the expression of Caspase 3, P53, PARP-1 and FAS and reduced the expression of Bcl-2 and increased the Bax/Bcl-2 ratio, nevertheless in none of the studied doses and times, had not a significant effect on the expression of Bax, Caspase 8 and Caspase 9. Conclusion: This study indicates that Naringenin can reduce the growth of some cancer cells and cause their deaths through increased apoptosis and decreased anti-apoptotic Bcl-2 gene expression and, resulting in the induction of apoptosis via both internal and external pathways.

Keywords: apoptosis, breast cancer, naringenin, T47D cell line

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4753 Rapid and Easy Fabrication of Collagen-Based Biocomposite Scaffolds for 3D Cell Culture

Authors: Esra Turker, Umit Hakan Yildiz, Ahu Arslan Yildiz

Abstract:

The key of regenerative medicine is mimicking natural three dimensional (3D) microenvironment of tissues by utilizing appropriate biomaterials. In this study, a synthetic biodegradable polymer; poly (L-lactide-co-ε-caprolactone) (PLLCL) and a natural polymer; collagen was used to mimic the biochemical structure of the natural extracellular matrix (ECM), and by means of electrospinning technique the real physical structure of ECM has mimicked. PLLCL/Collagen biocomposite scaffolds enables cell attachment, proliferation and nutrient transport through fabrication of micro to nanometer scale nanofibers. Biocomposite materials are commonly preferred due to limitations of physical and biocompatible properties of natural and synthetic materials. Combination of both materials improves the strength, degradation and biocompatibility of scaffold. Literature studies have shown that collagen is mostly solved with heavy chemicals, which is not suitable for cell culturing. To overcome this problem, a new approach has been developed in this study where polyvinylpyrrolidone (PVP) is used as co-electrospinning agent. PVP is preferred due to its water solubility, so PLLCL/collagen biocomposite scaffold can be easily and rapidly produced. Hydrolytic and enzymatic biodegradation as well as mechanical strength of scaffolds were examined in vitro. Cell adhesion, proliferation and cell morphology characterization studies have been performed as well. Further, on-chip drug screening analysis has been performed over 3D tumor models. Overall, the developed biocomposite scaffold was used for 3D tumor model formation and obtained results confirmed that developed model could be used for drug screening studies to predict clinical efficacy of a drug.

Keywords: biomaterials, 3D cell culture, drug screening, electrospinning, lab-on-a-chip, tissue engineering

Procedia PDF Downloads 294
4752 The Role of Cyfra 21-1 in Diagnosing Non Small Cell Lung Cancer (NSCLC)

Authors: H. J. T. Kevin Mozes, Dyah Purnamasari

Abstract:

Background: Lung cancer accounted for the fourth most common cancer in Indonesia. 85% of all lung cancer cases are the Non-Small Cell Lung Cancer (NSCLC). The indistinct signs and symptoms of NSCLC sometimes lead to misdiagnosis. The gold standard assessment for the diagnosis of NSCLC is the histopathological biopsy, which is invasive. Cyfra 21-1 is a tumor marker, which can be found in the intermediate protein structure in the epitel. The accuracy of Cyfra 21-1 in diagnosing NSCLC is not yet known, so this report is made to seek the answer for the question above. Methods: Literature searching is done using online databases. Proquest and Pubmed are online databases being used in this report. Then, literature selection is done by excluding and including based on inclusion criterias and exclusion criterias. The selected literature is then being appraised using the criteria of validity, importance, and validity. Results: From six journals appraised, five of them are valid. Sensitivity value acquired from all five literature is ranging from 50-84.5 %, meanwhile the specificity is 87.8 %-94.4 %. Likelihood the ratio of all appraised literature is ranging from 5.09 -10.54, which categorized to Intermediate High. Conclusion: Serum Cyfra 21-1 is a sensitive and very specific tumor marker for diagnosis of non-small cell lung cancer (NSCLC).

Keywords: cyfra 21-1, diagnosis, nonsmall cell lung cancer, NSCLC, tumor marker

Procedia PDF Downloads 218
4751 A Study of Blood Alcohol Concentration in People Arrested for Various Offences and Its Demographic Pattern

Authors: Tabin Millo, Khoob Chand, Ashok Kumar Jaiswal

Abstract:

Introduction: Various kinds of violence and offences are related to alcohol consumption by the offenders. The relationship between alcohol and violence is complex. But its study is important to achieve understanding of violence as well as alcohol related behavior. This study was done to know the blood alcohol concentration in people involved in various offences and its demographic pattern. The study was carried out in the forensic toxicology laboratory, department of Forensic Medicine, All India Institute of Medical Sciences, New Delhi, India. Material and methods: The blood samples were collected from the arrested people shortly after the commission of the offence by the emergency medical officers in the emergency department and forwarded to the forensic toxicology laboratory through the investigating officer. The blood samples were collected in EDTA vial with sodium fluoride preservative. The samples were analyzed by using gas chromatography with head space (GC-HS), which is ideal for alcohol estimation. The toxicology reports were given within a week. The data of seven years (2011-17) were analyzed for its alcohol concentration, associated crimes and its demographic pattern. Analysis and conclusion: Total 280 samples were analyzed in the period of 2011-2017. All were males except one female who was a bar dancer. The maximum cases were in the age group of 21-30 years (124 cases). The type of offences involved were road traffic accidents (RTA), assault cases, drunken driving, drinking in public place, drunk on duty, sexual offence, bestiality, eve teasing, fall etc. The maximum cases were of assault (75 cases) followed by RTA (64 cases). The maximum cases were in the alcohol concentration range of 101-150mg% (58 cases) followed by 51-100mg% (52 cases). The maximum blood alcohol level detected was 391.51 mg%, belonging to a security guard found unconscious. This study shows that alcohol consumption is associated with various kinds of violence and offences in society.

Keywords: alcohol, crime, toxicology, violence

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4750 Ellagic Acid Enhanced Apoptotic Radiosensitivity via G1 Cell Cycle Arrest and γ-H2AX Foci Formation in HeLa Cells in vitro

Authors: V. R. Ahire, A. Kumar, B. N. Pandey, K. P. Mishra, G. R. Kulkarni

Abstract:

Radiation therapy is an effective vital strategy used globally in the treatment of cervical cancer. However, radiation efficacy principally depends on the radiosensitivity of the tumor, and not all patient exhibit significant response to irradiation. A radiosensitive tumor is easier to cure than a radioresistant tumor which later advances to local recurrence and metastasis. Herbal polyphenols are gaining attention for exhibiting radiosensitization through various signaling. Current work focuses to study the radiosensitization effect of ellagic acid (EA), on HeLa cells. EA intermediated radiosensitization of HeLa cells was due to the induction γ-H2AX foci formation, G1 phase cell cycle arrest, and loss of reproductive potential, growth inhibition, drop in the mitochondrial membrane potential and protein expression studies that eventually induced apoptosis. Irradiation of HeLa in presence of EA (10 μM) to doses of 2 and 4 Gy γ-radiation produced marked tumor cytotoxicity. EA also demonstrated radio-protective effect on normal cell, NIH3T3 and aided recovery from the radiation damage. Our results advocate EA to be an effective adjuvant for improving cancer radiotherapy as it displays striking tumor cytotoxicity and reduced normal cell damage instigated by irradiation.

Keywords: apoptotic radiosensitivity, ellagic acid, mitochondrial potential, cell-cycle arrest

Procedia PDF Downloads 335
4749 Evaluating the Performance of Passive Direct Methanol Fuel Cell under Varying Operating and Structural Conditions

Authors: Rahul Saraswat

Abstract:

More recently, a focus has been given to replacing machined stainless steel metal flow fields with inexpensive wire mesh current collectors. The flow fields are based on simple woven wire mesh screens of various stainless steels, which are sandwiched between a thin metal plate of the same material to create a bipolar plate/flow field configuration for use in a stack. Major advantages of using stainless steel wire screens include the elimination of expensive raw materials as well as machining and/or other special fabrication costs. The objective of the project is to improve the performance of the passive direct methanol fuel cell without increasing the cost of the cell and to make it as compact and light as possible. From the literature survey, it was found that very little is done in this direction, and the following methodology was used. 1. The passive direct methanol fuel cell (DMFC) can be made more compact, lighter, and less costly by changing the material used in its construction. 2. Controlling the fuel diffusion rate through the cell improves the performance of the cell. A passive liquid feed direct methanol fuel cell (DMFC) was fabricated using a given MEA (Membrane Electrode Assembly) and tested for different current collector structures. Mesh current collectors of different mesh densities along with different support structures, were used, and the performance was found to be better. Methanol concentration was also varied. Optimisation of mesh size, support structure, and fuel concentration was achieved. Cost analysis was also performed hereby. From the performance analysis study of DMFC, we can conclude with the following points: Area specific resistance (ASR) of wire mesh current collectors is lower than the ASR of stainless steel current collectors. Also, the power produced by wire mesh current collectors is always more than that produced by stainless steel current collectors. 1. Low or moderate methanol concentrations should be used for better and stable DMFC performance. 2. Wiremesh is a good substitute for stainless steel for current collector plates of passive DMFC because of its lower cost (by about 27 %), flexibility, and light in weight characteristics of wire mesh.

Keywords: direct methanol fuel cell, membrane electrode assembly, mesh, mesh size, methanol concentration, support structure

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4748 COX-2 Inhibitor NS398 Counteracts Chemoresistance to Temozolomide in T98G Glioblastoma Cell Line

Authors: Francesca Lombardi, Francesca Rosaria Augello, Benedetta Cinque, Maria Grazia Cifone, Paola Palumbo

Abstract:

Glioblastoma multiforme (GBM) is a high-grade primary brain tumor refractory to current forms of treatment. The survival benefits of patients with GBM remain unsatisfactory due to the intrinsic or acquired resistance to temozolomide (TMZ), an alkylating agent, used as the first-line chemotherapeutic drug to treat GBM patients. Its cytotoxic effect is visualized by the induction of O6-methylguanine (O6MeG) within DNA. Cyclooxygenase-2 (COX-2), an inflammation-associated enzyme, has been implicated in tumorigenesis and progression of GBM, its inhibition shows anticancer activities. In the present study, it was verified if the combination of a COX-2 selective inhibitor, NS398, with TMZ could counteract the TMZ resistance. In particular, the effect of NS398 mixed with TMZ was investigated in the GBM TMZ-resistant cell line, T98G. Cells were pretreated with NS398 (100µM, 24 hours) and then exposed to TMZ alone (200µM), NS398 alone, or both for 72 hours, after which cell growth rate and cycle phases, as well as apoptosis level, were evaluated. Coadministration of NS398 and TMZ caused a significant decrease in cell growth and a progressive increase of dead cells detected by trypan blue staining. Moreover, a significant level of apoptotic cell percentage and alteration of cell cycle phases were observed in T98G treated with TMZ-NS398 combination when compared to untreated cells or TMZ-treated cells. TMZ-resistant tumors, as GBM, express elevated levels of DNA repair enzyme O6-methylguanine-DNA methyltransferase (MGMT). The mixture drastically reduced MGMT expression in the TMZ-resistant cell line T98G, known to express high levels of MGMT basically. Moreover, while TMZ alone did not influence the COX-2 protein expression, the combination successfully reduced it. In conclusion, these results demonstrated that NS398 enhanced the efficacy of TMZ through cell number reduction, apoptosis induction, and decreased MGMT levels, suggesting the ability of drug combination to reduce the chemoresistance. This drug combination deserves attention and could be considered as a promising therapeutic strategy for GBM patients.

Keywords: COX-2, COX-2 inhibitor, glioblastoma, NS398, T98G, temozolomide

Procedia PDF Downloads 129
4747 Prevalence and Determinants of Hypertension among the Santal Indigenous Group in Bangladesh

Authors: Sharmin Sultana, Palash Chandra Banik, Shirin Jahan Mumu, Liaquat Ali

Abstract:

Santals are one of the oldest indigenous groups of South Asia who, according to anthropological evidence, are thought to be the origins of the Bengali race. The aim of the study was to explore, according to our best knowledge for the first time, the prevalence and determinants of hypertension in this relatively isolated and marginalized indigenous group who still live mostly in a traditional style. Under a cross-sectional analytical design, the study was conducted on the adult (age≥18 years) Santals (n=389, M/F 184/205, age in years, 38±15.3) of a village located in a remote rural area of northern Bangladesh. Subjects were selected by purposive sampling, and data were collected by interviewer-administered pretested questionnaire. Blood pressure was measured by following the WHO guideline of JNC-7 has been used to classify the blood pressure. The prevalence of hypertension was 4.9% among the respondents. Females had a much higher prevalence (5.4%) of hypertension compared to males (4.3%). Among the risk indicators of hypertension, more than half (50.9%) of the study population took extra salt in their meals, whereas 10.5% of respondents used extra salt occasionally, which is an important risk factor for high blood pressure. High waist circumference was found in 19% of the study subjects in terms of central obesity. Older age group (p=0.003, OR=1.1, 95%CI-1.02-1.10), respondents who completed more than primary school (p=0.038, OR=7.1, CI-1.11, 44.6), overweight and obesity (p=0.004, OR=17.1, CI-2.5, 118.1), were the major determinant for hypertension as found from the binary logistic model. None of the respondents received any medication, neither they visit any doctor ever for their hypertension control. The prevalence of hypertension was found to be low but not ignorable. Pre-hypertension in the case of systolic blood pressure needs attention among Santal indigenous population.

Keywords: hypertension, indigenous group, Santals, Bangladesh

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4746 Management and Evaluating Technologies of Tissue Engineering Various Fields of Bone

Authors: Arash Sepehri Bonab

Abstract:

Techniques to switch cells between development and differentiation, which tend to be commonly exclusive, are utilized in arrange to supply an expansive cell mass that can perform particular separated capacities required for the tissue to develop. Approaches to tissue engineering center on the have to give signals to cell populaces to advance cell multiplication and separation. Current tissue regenerative procedures depend primarily on tissue repair by transplantation of synthetic/natural inserts. In any case, restrictions on the existing procedures have expanded the request for tissue designing approaches. Tissue engineering innovation and stem cell investigation based on tissue building have made awesome advances in overcoming the issues of tissue and organ damage, useful loss, and surgical complications. Bone tissue has the capability to recover itself; in any case, surrenders of a basic estimate anticipate the bone from recovering and require extra support. The advancement of bone tissue building has been utilized to form useful options to recover the bone. This paper primarily portrays current advances in tissue engineering in different fields of bone and talks about the long-term trend of tissue designing innovation in the treatment of complex diseases.

Keywords: tissue engineering, bone, technologies, treatment

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4745 Synthesis of [1-(Substituted-Sulfonyl)-Piperidin-4-yl]-(2,4-Difluoro-Phenyl)-Methanone Oximes and Their Biological Activity

Authors: L. Mallesha, C. S. Karthik, P. Mallu

Abstract:

A series of new [1-(substituted-benzoyl)-piperidin-4-yl]-(2,4-difluoro-phenyl)-methanone oxime derivatives, 3(a-f) were synthesized and characterized by different spectral studies. All compounds were evaluated for their in vitro antibacterial activity against bacterial strains. These compounds were screened for their antioxidant activity by DPPH• and Fe2+ chelating assay. Antiproliferative effects were evaluated using the MTT assay method against two human cancer cell lines and one astrocytoma brain tumor cell line. Compound 3b exhibited moderate antibacterial activity when compared with other compounds. All the compounds showed antioxidant activity, where compound 3f was the best radical scavenger and Fe2+ ion scavenger. Compounds, 3b, and 3d showed good activity on all cell lines, whereas the other compounds in the series exhibited moderate activity.

Keywords: Piperidine, antibacterial, antioxidant, antiproliferative

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4744 Prolonged Synthesis of Chitin Polysaccharide from Chlorovirus System

Authors: Numfon Rakkhumkaew, Takeru Kawasaki, Makoto Fujie, Takashi Yamada

Abstract:

Chlorella viruses or chloroviruses contain a gene that encodes a function for chitin synthesis, which is expressed early in viral infection to produce chitin polysaccharide, a polymer of β-1, 4-linked GlcNAc, on the outside of Chlorella cell wall. Interestingly, chlorovirus system is an eco-friendly system which converses CO2 and solar energy from the environment into useful materials. However, infected Chlorella cells are lysed at the final stage of viral infection, and this phenomenon is caused the breaking down of polysaccharide. To postpone the lysing period and prolong the synthesis of chitin polysaccharide on cells, the slow growing virus incorporated with aphidicolin treatment, an inhibitor of DNA synthesis, was investigated. In this study, a total of 25 virus isolates from water samples in Japan region were analyzed for CHS (the gene for CH synthase) gene by PCR (polymerase chain reaction). The accumulation and appearance of chitin polysaccharide on infected cells were detected by biotinylated chitin-binding proteins WGA (wheat germ agglutinin)-biotin for chitin in conjunction with avidin-Cy 2 or Cy 3 and investigated by fluorescence microscopy, observed as green or yellow fluorescence over the cell surface. Among all chlorovirus isolates, cells infected with CNF1 revealed the accumulation of chitin over the cell surface within 30 min p.i. and continued to accumulate on cells until 4 h p.i. before cell lyses which was 1.6 times longer accumulation period than cells infected with CVK2 (prototype virus). Furthermore, addition of aphidicolin could extend the chitin accumulation on cells infected with CNF1 until 8 h p.i. before cell lyses. Whereas, CVK2-infected cells treated with aphidicolin could prolong the chitin synthesis only for 6 h p.i. before cell lyses. Therefore, chitin synthesis by Chlorella-virus system could be prolonged by using slow-growing viral isolates and with aphidicolin.

Keywords: chitin, chlorovirus, Chlorella virus, aphidicolin

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4743 An Unbiased Profiling of Immune Repertoire via Sequencing and Analyzing T-Cell Receptor Genes

Authors: Yi-Lin Chen, Sheng-Jou Hung, Tsunglin Liu

Abstract:

Adaptive immune system recognizes a wide range of antigens via expressing a large number of structurally distinct T cell and B cell receptor genes. The distinct receptor genes arise from complex rearrangements called V(D)J recombination, and constitute the immune repertoire. A common method of profiling immune repertoire is via amplifying recombined receptor genes using multiple primers and high-throughput sequencing. This multiplex-PCR approach is efficient; however, the resulting repertoire can be distorted because of primer bias. To eliminate primer bias, 5’ RACE is an alternative amplification approach. However, the application of RACE approach is limited by its low efficiency (i.e., the majority of data are non-regular receptor sequences, e.g., containing intronic segments) and lack of the convenient tool for analysis. We propose a computational tool that can correctly identify non-regular receptor sequences in RACE data via aligning receptor sequences against the whole gene instead of only the exon regions as done in all other tools. Using our tool, the remaining regular data allow for an accurate profiling of immune repertoire. In addition, a RACE approach is improved to yield a higher fraction of regular T-cell receptor sequences. Finally, we quantify the degree of primer bias of a multiplex-PCR approach via comparing it to the RACE approach. The results reveal significant differences in frequency of VJ combination by the two approaches. Together, we provide a new experimental and computation pipeline for an unbiased profiling of immune repertoire. As immune repertoire profiling has many applications, e.g., tracing bacterial and viral infection, detection of T cell lymphoma and minimal residual disease, monitoring cancer immunotherapy, etc., our work should benefit scientists who are interested in the applications.

Keywords: immune repertoire, T-cell receptor, 5' RACE, high-throughput sequencing, sequence alignment

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4742 The Evaluation for Interfacial Adhesion between SOFC and Metal Adhesive in the High Temperature Environment

Authors: Sang Koo Jeon, Seung Hoon Nahm, Oh Heon Kwon

Abstract:

The unit cell of solid oxide fuel cell (SOFC) must be stacked as several layers type to obtain the high power. The most of researcher have concerned about the performance of stacked SOFC rather than the structural stability of stacked SOFC and especially interested how to design for reducing the electrical loss and improving the high efficiency. Consequently, the stacked SOFC able to produce the electrical high power and related parts like as manifold, gas seal, bipolar plate were developed to optimize the stack design. However, the unit cell of SOFC was just layered on the interconnector without the adhesion and the hydrogen and oxygen were injected to the interfacial layer in the high temperature. On the operating condition, the interfacial layer can be the one of the weak point in the stacked SOFC. Therefore the evaluation of the structural safety for the failure is essentially needed. In this study, interfacial adhesion between SOFC and metal adhesive was estimated in the high temperature environment. The metal adhesive was used to strongly connect the unit cell of SOFC with interconnector and provide the electrical conductivity between them. The four point bending test was performed to measure the interfacial adhesion. The unit cell of SOFC and SiO2 wafer were diced and then attached by metal adhesive. The SiO2 wafer had the center notch to initiate a crack from the tip of the notch. The modified stereomicroscope combined with the CCD camera and system for measuring the length was used to observe the fracture behavior. Additionally, the interfacial adhesion was evaluated in the high temperature condition because the metal adhesive was affected by high temperature. Also the specimen was exposed in the furnace during several hours and then the interfacial adhesion was evaluated. Finally, the interfacial adhesion energy was quantitatively determined and compared in the each condition.

Keywords: solid oxide fuel cell (SOFC), metal adhesive, adhesion, high temperature

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