Search results for: cell wall biosynthesis
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 4826

Search results for: cell wall biosynthesis

4076 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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4075 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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4074 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

Procedia PDF Downloads 152
4073 Comparative Mesh Sensitivity Study of Different Reynolds Averaged Navier Stokes Turbulence Models in OpenFOAM

Authors: Zhuoneng Li, Zeeshan A. Rana, Karl W. Jenkins

Abstract:

In industry, to validate a case, often a multitude of simulation are required and in order to demonstrate confidence in the process where users tend to use a coarser mesh. Therefore, it is imperative to establish the coarsest mesh that could be used while keeping reasonable simulation accuracy. To date, the two most reliable, affordable and broadly used advanced simulations are the hybrid RANS (Reynolds Averaged Navier Stokes)/LES (Large Eddy Simulation) and wall modelled LES. The potentials in these two simulations will still be developed in the next decades mainly because the unaffordable computational cost of a DNS (Direct Numerical Simulation). In the wall modelled LES, the turbulence model is applied as a sub-grid scale model in the most inner layer near the wall. The RANS turbulence models cover the entire boundary layer region in a hybrid RANS/LES (Detached Eddy Simulation) and its variants, therefore, the RANS still has a very important role in the state of art simulations. This research focuses on the turbulence model mesh sensitivity analysis where various turbulence models such as the S-A (Spalart-Allmaras), SSG (Speziale-Sarkar-Gatski), K-Omega transitional SST (Shear Stress Transport), K-kl-Omega, γ-Reθ transitional model, v2f are evaluated within the OpenFOAM. The simulations are conducted on a fully developed turbulent flow over a flat plate where the skin friction coefficient as well as velocity profiles are obtained to compare against experimental values and DNS results. A concrete conclusion is made to clarify the mesh sensitivity for different turbulence models.

Keywords: mesh sensitivity, turbulence models, OpenFOAM, RANS

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4072 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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4071 Prevalence of Dengue in Sickle Cell Disease in Pre-school Children

Authors: Nikhil A. Gavhane, Sachin Shah, Ishant S. Mahajan, Pawan D. Bahekar

Abstract:

Introduction: Millions of people are affected with dengue fever every year, which drives up healthcare expenses in many low-income countries. Organ failure and other serious symptoms may result. Another worldwide public health problem is sickle cell anaemia, which is most prevalent in Africa, the Caribbean, and Europe. Dengue epidemics have reportedly occurred in locations with a high frequency of sickle cell disease, compounding the health problems in these areas. Aims and Objectives: This study examines dengue infection in sickle cell disease-afflicted pre-schoolers. Method:This Retrospective cohort study examined paediatric patients. Young people with sickle cell disease (SCD), dengue infection, and a control group without SCD or dengue were studied. Data on demographics, SCD consequences, medical treatments, and laboratory findings were gathered to analyse the influence of SCD on dengue severity and clinical outcomes, classified as severe or non-severe by the 2009 WHO classification. Using fever or admission symptoms, the research estimated acute illness duration. Result: Table 1 compares haemoglobin genotype-based dengue episode features in SS, SC, and controls. Table 2 shows that severe dengue cases are older, have longer admission delays, and have particular symptoms. Table 3's multivariate analysis indicates SS genotype's high connection with severe dengue, multiorgan failure, and acute pulmonary problems. Table 4 relates severe dengue to greater white blood cell counts, anaemia, liver enzymes, and reduced lactate dehydrogenase. Conclusion: This study is valuable but confined to hospitalised dengue patients with sickle cell illness. Small cohorts limit comparisons. Further study is needed since findings contradict predictions.

Keywords: dengue, chills, headache, severe myalgia, vomiting, nausea, prostration

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4070 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

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4069 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

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4068 Leukocyte Detection Using Image Stitching and Color Overlapping Windows

Authors: Lina, Arlends Chris, Bagus Mulyawan, Agus B. Dharmawan

Abstract:

Blood cell analysis plays a significant role in the diagnosis of human health. As an alternative to the traditional technique conducted by laboratory technicians, this paper presents an automatic white blood cell (leukocyte) detection system using Image Stitching and Color Overlapping Windows. The advantage of this method is to present a detection technique of white blood cells that are robust to imperfect shapes of blood cells with various image qualities. The input for this application is images from a microscope-slide translation video. The preprocessing stage is performed by stitching the input images. First, the overlapping parts of the images are determined, then stitching and blending processes of two input images are performed. Next, the Color Overlapping Windows is performed for white blood cell detection which consists of color filtering, window candidate checking, window marking, finds window overlaps, and window cropping processes. Experimental results show that this method could achieve an average of 82.12% detection accuracy of the leukocyte images.

Keywords: color overlapping windows, image stitching, leukocyte detection, white blood cell detection

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4067 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

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4066 Assessing Overall Thermal Conductance Value of Low-Rise Residential Home Exterior Above-Grade Walls Using Infrared Thermography Methods

Authors: Matthew D. Baffa

Abstract:

Infrared thermography is a non-destructive test method used to estimate surface temperatures based on the amount of electromagnetic energy radiated by building envelope components. These surface temperatures are indicators of various qualitative building envelope deficiencies such as locations and extent of heat loss, thermal bridging, damaged or missing thermal insulation, air leakage, and moisture presence in roof, floor, and wall assemblies. Although infrared thermography is commonly used for qualitative deficiency detection in buildings, this study assesses its use as a quantitative method to estimate the overall thermal conductance value (U-value) of the exterior above-grade walls of a study home. The overall U-value of exterior above-grade walls in a home provides useful insight into the energy consumption and thermal comfort of a home. Three methodologies from the literature were employed to estimate the overall U-value by equating conductive heat loss through the exterior above-grade walls to the sum of convective and radiant heat losses of the walls. Outdoor infrared thermography field measurements of the exterior above-grade wall surface and reflective temperatures and emissivity values for various components of the exterior above-grade wall assemblies were carried out during winter months at the study home using a basic thermal imager device. The overall U-values estimated from each methodology from the literature using the recorded field measurements were compared to the nominal exterior above-grade wall overall U-value calculated from materials and dimensions detailed in architectural drawings of the study home. The nominal overall U-value was validated through calendarization and weather normalization of utility bills for the study home as well as various estimated heat loss quantities from a HOT2000 computer model of the study home and other methods. Under ideal environmental conditions, the estimated overall U-values deviated from the nominal overall U-value between ±2% to ±33%. This study suggests infrared thermography can estimate the overall U-value of exterior above-grade walls in low-rise residential homes with a fair amount of accuracy.

Keywords: emissivity, heat loss, infrared thermography, thermal conductance

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4065 Phytosynthesized Iron Nanoparticles Elicited Growth and Biosynthesis of Steviol Glycosides in Invitro Stevia rebaudiana Plant Cultures

Authors: Amir Ali, Laura Yael Mendoza

Abstract:

The application of nanomaterials is becoming the most effective strategy of elicitation to produce a desirable level of plant biomass with complex medicinal compounds. This study was designed to check the influence of phytosynthesized iron nanoparticles (FeNPs) on physical growth characteristics, antioxidant status, and production of steviol glycosides of in vitro grown Stevia rebaudiana. Effect of different concentrations of iron nanoparticles replacement of iron sulfate in MS medium (stock solution) on invitro stevia plant growth following positive control (MS basal medium), negative control (iron sulfate devoid medium), iron sulfate devoid MS medium and supplemented with FeNPs at different concentrations (5.6 mg/L, 11.2 mg/L, 16.8 mg/L, 22.4 mg/L) was evaluated. The iron deficiency leads to a drastic reduction in plant growth. In contrast, applying FeNPs leads to improvement in plant height, leave diameter, improved leave morphology, etc., in a concentration-dependent manner. Furthermore, the stress caused by FeNPs at 16.8 mg/L in cultures produced higher levels of total phenolic content (3.7 ± 0.042 mg/g dry weight: DW) and total flavonoid content (1.9 ± 0.022 mg/g DW and antioxidant activity (78 ± 4.6%). In addition, plants grown in the presence of FeNPs at 22.4 mg/L resulted in higher enzymatic antioxidant activities (SOD = 3.5 ± 0.042 U/mg; POD = 2.6 ± 0.026 U/mg; CAT = 2.8 ± 0.034 U/mg and APx = 3.6 ± 0.043 U/ mg), respectively. Furthermore, exposure to a higher dose of FeNPs (22.4 mg/L) exhibited the maximum amount of stevioside (stevioside: 4.6 ± 0.058 mg/g (DW) and rebaudioside A: 4.9 ± 0.068 mg/g DW) as compared to other doses. The current investigation confirms the effectiveness of FeNPs in growth media. It offers a suitable prospect for commercially desirable production of S. rebaudiana biomass with higher sweet glycosides profiles in vitro.

Keywords: cell culture, stevia, iron nanoparticles, antioxidants

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4064 Unzipping the Stress Response Genes in Moringa oleifera Lam. through Transcriptomics

Authors: Vivian A. Panes, Raymond John S. Rebong, Miel Q. Diaz

Abstract:

Moringa oleifera Lam. is known mainly for its high nutritional value and medicinal properties contributing to its popular reputation as a 'miracle plant' in the tropical climates where it usually grows. The main objective of this study is to discover the genes and gene products involved in abiotic stress-induced activity that may impact the M. oleifera Lam. mature seeds as well as their corresponding functions. In this study, RNA-sequencing and de novo transcriptome assembly were performed using two assemblers, Trinity and Oases, which produced 177,417 and 120,818 contigs respectively. These transcripts were then subjected to various bioinformatics tools such as Blast2GO, UniProt, KEGG, and COG for gene annotation and the analysis of relevant metabolic pathways. Furthermore, FPKM analysis was performed to identify gene expression levels. The sequences were filtered according to the 'response to stress' GO term since this study dealt with stress response. Clustered Orthologous Groups (COG) showed that the highest frequencies of stress response gene functions were those of cytoskeleton which make up approximately 14% and 23% of stress-related sequences under Trinity and Oases respectively, recombination, repair and replication at 11% and 14% respectively, carbohydrate transport and metabolism at 23% and 9% respectively and defense mechanisms 16% and 12% respectively. KEGG pathway analysis determined the most abundant stress-response genes in the phenylpropanoid biosynthesis at counts of 187 and 166 pathways for Oases and Trinity respectively, purine metabolism at 123 and 230 pathways, and biosynthesis of antibiotics at 105 and 102. Unique and cumulative GO term counts revealed that majority of the stress response genes belonged to the category of cellular response to stress at cumulative counts of 1,487 to 2,187 for Oases and Trinity respectively, defense response at 754 and 1,255, and response to heat at 213 and 208, response to water deprivation at 229 and 228, and oxidative stress at 508 and 488. Lastly, FPKM was used to determine the levels of expression of each stress response gene. The most upregulated gene encodes for thiamine thiazole synthase chloroplastic-like enzyme which plays a significant role in DNA damage tolerance. Data analysis implies that M. oleifera stress response genes are directed towards the effects of climate change more than other stresses indicating the potential of M. oleifera for cultivation in harsh environments because it is resistant to climate change, pathogens, and foreign invaders.

Keywords: stress response, genes, Moringa oleifera, transcriptomics

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4063 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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4062 Understanding the Role of Nitric Oxide Synthase 1 in Low-Density Lipoprotein Uptake by Macrophages and Implication in Atherosclerosis Progression

Authors: Anjali Roy, Mirza S. Baig

Abstract:

Atherosclerosis is a chronic inflammatory disease characterized by the formation of lipid rich plaque enriched with necrotic core, modified lipid accumulation, smooth muscle cells, endothelial cells, leucocytes and macrophages. Macrophage foam cells play a critical role in the occurrence and development of inflammatory atherosclerotic plaque. Foam cells are the fat-laden macrophages in the initial stage atherosclerotic lesion formation. Foam cells are an indication of plaque build-up, or atherosclerosis, which is commonly associated with increased risk of heart attack and stroke as a result of arterial narrowing and hardening. The mechanisms that drive atherosclerotic plaque progression remain largely unknown. Dissecting the molecular mechanism involved in process of macrophage foam cell formation will help to develop therapeutic interventions for atherosclerosis. To investigate the mechanism, we studied the role of nitric oxide synthase 1(NOS1)-mediated nitric oxide (NO) on low-density lipoprotein (LDL) uptake by bone marrow derived macrophages (BMDM). Using confocal microscopy, we found that incubation of macrophages with NOS1 inhibitor, TRIM (1-(2-Trifluoromethylphenyl) imidazole) or L-NAME (N omega-nitro-L-arginine methyl ester) prior to LDL treatment significantly reduces the LDL uptake by BMDM. Further, addition of NO donor (DEA NONOate) in NOS1 inhibitor treated macrophages recovers the LDL uptake. Our data strongly suggest that NOS1 derived NO regulates LDL uptake by macrophages and foam cell formation. Moreover, we also checked proinflammatory cytokine mRNA expression through real time PCR in BMDM treated with LDL and copper oxidized LDL (OxLDL) in presences and absences of inhibitor. Normal LDL does not evoke cytokine expression whereas OxLDL induced proinflammatory cytokine expression which significantly reduced in presences of NOS1 inhibitor. Rapid NOS-1-derived NO and its stable derivative formation act as signaling agents for inducible NOS-2 expression in endothelial cells, leading to endothelial vascular wall lining disruption and dysfunctioning. This study highlights the role of NOS1 as critical players of foam cell formation and would reveal much about the key molecular proteins involved in atherosclerosis. Thus, targeting NOS1 would be a useful strategy in reducing LDL uptake by macrophages at early stage of disease and hence dampening the atherosclerosis progression.

Keywords: atherosclerosis, NOS1, inflammation, oxidized LDL

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4061 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

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4060 Second-Order Slip Flow and Heat Transfer in a Long Isothermal Microchannel

Authors: Huei Chu Weng, Chien-Hung Liu

Abstract:

This paper presents a study on the effect of second-order slip and jump on forced convection through a long isothermally heated or cooled planar microchannel. The fully developed solutions of thermal flow fields are analytically obtained on the basis of the second-order Maxwell-Burnett slip and Smoluchowski jump boundary conditions. Results reveal that the second-order term in the Karniadakis slip boundary condition is found to contribute a negative velocity slip and then to lead to a higher pressure drop as well as a higher fluid temperature for the heated-wall case or to a lower fluid temperature for the cooled-wall case. These findings are contrary to predictions made by the Deissler model. In addition, the role of second-order slip becomes more significant when the Knudsen number increases.

Keywords: microfluidics, forced convection, gas rarefaction, second-order boundary conditions

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4059 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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4058 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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4057 Nonlinear Flow Behavior and Validity of the Cubic Law in a Rough Fracture

Authors: Kunwar Mrityunjai Sharma, Trilok Nath Singh

Abstract:

The Navier-Stokes equation is used to study nonlinear fluid flow in rough 2D fractures. The major goal is to investigate the influence of inertial flow owing to fracture wall roughness on nonlinear flow behavior. Roughness profiles are developed using Barton's Joint Roughness Coefficient (JRC) and used as fracture walls to assess wall roughness. Four JRC profiles (5, 11, 15, and 19) are employed in the study, where a higher number indicates higher roughness. A parametric study has been performed using varying pressure gradients, and the corresponding Forchheimer number is calculated to observe the nonlinear behavior. The results indicate that the fracture roughness has a significant effect on the onset of nonlinearity. Additionally, the validity of the cubic law is evaluated and observed that it overestimates the flow in rough fractures and should be used with utmost care.

Keywords: fracture flow, nonlinear flow, cubic law, Navier-stokes equation

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4056 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

Procedia PDF Downloads 176
4055 Antimicrobial Action and Its Underlying Mechanism by Methanolic Seed Extract of Syzygium cumini on Bacillus subtilis

Authors: Alok Kumar Yadav, Saurabh Saraswat, Preeti Sirohi, Manjoo Rani, Sameer Srivastava, Manish Pratap Singh, Nand K. Singh

Abstract:

The development of antibiotic resistance in bacteria is increasing at an alarming rate, and this is considered as one of the most serious threats in the history of medicine, and an alternative solution should be derived so as to tackle this problem. In many countries, people use the medicinal plants for the treatment of various diseases as these are cheaper, easily available and least toxic. Syzygium cumini is used for the treatment of various kinds of diseases but their mechanism of action is not reported. The antimicrobial activity of Syzygium cumini was tested by the well diffusion assay and zone of inhibition was reported to be 20.06 mm as compared to control with MIC of 0.3 mg/ml. Genomic DNA fragmentation of Bacillus subtilis revealed apoptosis and FE-SEM indicate cell wall cracking on several intervals of time. Propidium iodide staining results showed that few bacterial cells were stained in the control and population of stained cells increase after exposing them for various period of time. Flow cytometric kinetic data analysis on the membrane permeabilization in bacterial cell showed the significant contribution of antimicrobial potential of the seed extract on antimicrobial-induced permeabilization. Two components of Syzygium cumini methanolic seed extract was found to be quite active against four enzymes like PDB ID- 1W5D, 4OX3, 3MFD and 5E2F which have a very crucial role in membrane synthesis in Bacillus subtilis by in silico analysis. Through in silico analysis, lupeol showed highest binding energy for macromolecule 1W5D and 4OX3 whereas stigmasterol showed the highest binding energy for macromolecule 3MFD and 5E2F respectively. It showed that methanolic seed extract of Syzygium cumini can be used for the inhibition of foodborne infections caused by Bacillus subtilis and also as an alternative of prevalent antibiotics.

Keywords: antibiotics, Bacillus subtilis, inhibition, Syzygium cumini

Procedia PDF Downloads 178
4054 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

Procedia PDF Downloads 507
4053 Effects of New Anthraquinone Derivatives on Resistance Ovarian Cancer Cells and The Mechanism Investigation

Authors: Hui-Hsin Huang, Sheng-Tung Huang, Chi-Ming Lee, Chiao-Han Yen, Chun-Mao Lin

Abstract:

At initiation stage, there are no symptoms at initiation stage; however, at late stage, patients suffer symptoms as soon as ovarian cancer metastasis. Moreover, ovarian cancer cells are resistant to some anti-ovarian cancer drugs in clinical. Thus, it is very important to find an effective treatment for resistant ovarian cancer. Anthraquinone derivatives are able to induce DNA damage and lead to cell apoptosis, so several derivatives have been used for clinical application. Therefore, to explore more effective anti-ovarian cancer drugs, this study investigates the mechanism of three new anthraquinone compounds bearing different functional groups to camptothecin-resistance ovarian cell line A2780R2000. Cell viability was determined by MTT assay after treating A2780R2000 with the three new anthraquinone compounds. The results indicated that IC50 values are 33.44μM (Compound I), 25.77μM (Compound II) and 24.59μM (Compound III). Next, through cell cycle analysis, the results demonstrated that three new anthraquinone compounds not only induced A2780R2000 cell cycle arrest at early stage but also apoptosis at late stage. Besides, through apoptosis assay, the results indicated new anthraquinone compound induced apoptosis at late stage. Furthermore, the results of western blot show that the three new anthraquinone compounds lead to A2780R2000 apoptosis through intrinsic pathway. Theses results suggested that three new anthraquinone compounds may be potential new drugs for clinical cancer treatment in the future.

Keywords: anthraquinone, camptothecin, resistance, ovarian cancer

Procedia PDF Downloads 377
4052 Effect of Chlorophyll Concentration Variations from Extract of Papaya Leaves on Dye-Sensitized Solar Cell

Authors: Eka Maulana, Sholeh Hadi Pramono, Dody Fanditya, M. Julius

Abstract:

In this paper, extract of papaya leaves are used as a natural dye and combined by variations of solvent concentration applied on DSSC (Dye-Sensitized Solar Cell). Indonesian geographic located on the equator line occasions the magnitude of the potential to develop organic solar cells made from extracts of chlorophyll as a substitute for inorganic materials or synthetic dye on DSSC material. Dye serves as absorbing photons which are then converted into electrical energy. A conductive coated glass layer called TCO (Transparent Conductive Oxide) is used as a substrate of electrode. TiO2 nanoparticles as binding dye molecules, redox couple iodide/ tri-iodide as the electrolyte and carbon as the counter electrode in the DSSC are used. TiO2 nanoparticles, organic dyes, electrolytes and counter electrode are arranged and combined with the layered structure of the photo-catalyst absorption layer. Dye absorption measurements using a spectrophotometer at 200-800 nm light spectrum produces a total amount of chlorophyll 80.076 mg/l. The test cell at 7 watt LED light with 5000 lux luminescence were obtained Voc and Isc of 235.5 mV and 14 μA, respectively.

Keywords: DSSC (Dye-Sensitized Solar Cell), natural dye, chlorophyll, absorption

Procedia PDF Downloads 483
4051 Transvaginal Repair of Anterior Vaginal Wall Prolapse with Polyvinylidene Fluoride (PVDF) Mesh: An Alternative for Previously Restricted Materials

Authors: Mohammad-Javad Eslami, Mahtab Zargham, Farshad Gholipour, Mohammadreza Hajian, Katayoun Bakhtiari, Sakineh Hajebrahimi, Melina Eghbal, Ziba Farajzadegan

Abstract:

Introduction: To study the mid-term safety and functional outcomes of transvaginal anterior vaginal wall prolapse repair using polyvinylidene fluoride (PVDF) mesh (DynaMesh®-PR4) by the double trans-obturator technique (TOT). Methods: Between 2015 and 2020, we prospectively included women with symptomatic high-stage anterior vaginal wall prolapse with or without uterine prolapse or stress urinary incontinence (SUI) in the study. The patients underwent transvaginal repair of the prolapse using PVDF mesh in two medical centers. We followed all patients for at least 12 months. We recorded the characteristics of vaginal and sexual symptoms, urinary incontinence, and prolapse stage pre- and postoperatively using International Consultation on Incontinence Questionnaire-Vaginal Symptoms (ICIQ-VS), International Consultation on Incontinence Questionnaire-Urinary Incontinence-Short Form (ICIQ-UI-SF), and Pelvic Organ Prolapse Quantification (POP-Q) system, respectively. Results: One hundred eight women were included in the final analysis with a mean follow-up time of 34.5 ± 18.6 months. The anatomical success was achieved in 103 (95.4%) patients. There was a significant improvement in patients’ vaginal symptoms, urinary incontinence, and quality of life scores postoperatively (p < 0.0001). Only six patients (5.5%) had mesh extrusion, five of whom were managed successfully. The total rates of complications and de novo urinary symptoms were 21.3% and 7.4%, respectively. Significant pain was reported in 17 cases (15.7%). Conclusion: Our findings show that using PVDF mesh in the double TOT technique for anterior vaginal wall prolapse repair is a safe procedure with high anatomic and functional success rates and acceptable complication rates in mid-term follow-up.

Keywords: stress urinary incontinence (SU, incontinence questionnaire-vaginal symptoms (ICIQ-VS), polyvinylidene fluoride (PVDF) mes, double trans-obturator technique (TOT)

Procedia PDF Downloads 19
4050 Engineering Ligand-Free Biodegradable-Based Nanoparticles for Cell Attachment and Growth

Authors: Simone F. Medeiros, Isabela F. Santos, Rodolfo M. Moraes, Jaspreet K. Kular, Marcus A. Johns, Ram Sharma, Amilton M. Santos

Abstract:

Tissue engineering aims to develop alternatives to treat damaged tissues by promoting their regeneration. Its basic principle is to place cells on a scaffold capable of promoting cell functions, and for this purpose, polymeric nanoparticles have been successfully used due to the ability of some macro chains to mimic the extracellular matrix and influence cell functions. In general, nanoparticles require surface chemical modification to achieve cell adhesion, and recent advances in their synthesis include methods for modifying the ligand density and distribution onto nanoparticles surface. However, this work reports the development of biodegradable polymeric nanoparticles capable of promoting cellular adhesion without any surface chemical modification by ligands. Biocompatible and biodegradable nanoparticles based on poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBHV) were synthesized by solvent evaporation method. The produced nanoparticles were small in size (85 and 125 nm) and colloidally stable against time in aqueous solution. Morphology evaluation showed their spherical shape with small polydispersity. Human osteoblast-like cells (MG63) were cultured in the presence of PHBHV nanoparticles, and growth kinetics were compared to those grown on tissue culture polystyrene (TCPS). Cell attachment on non-tissue culture polystyrene (non-TCPS) pre-coated with nanoparticles was assessed and compared to attachment on TCPS. These findings reveal the potential of PHBHV nanoparticles for cell adhesion and growth, without requiring a matrix ligand to support cells, to be used as scaffolds, in tissue engineering applications.

Keywords: tissue engineering, PHBHV, stem cells, cellular attachment

Procedia PDF Downloads 196
4049 Mathematical Modeling of Skin Condensers for Domestic Refrigerator

Authors: Nitin Ghule, S. G. Taji

Abstract:

A mathematical model of hot-wall condensers used in refrigerators is presented. The model predicts the heat transfer characteristics of condenser and the effects of various design and operating parameters on condenser tube length and capacity. A finite element approach was used to model the condenser. The condenser tube is divided into elemental units, with each element consisting of adhesive tape, refrigerant tube and outer metal sheet. The heat transfer characteristics of each section are then analyzed by considering the heat transfer through the tube wall, tape and the outer sheet. Variations in inner heat transfer coefficient and pressure drop are considered depending on temperature, fluid phase, type of flow and orientation of tube. Variation in outer heat transfer coefficient is also taken into account. Various materials were analysed for the tube, tape and outer sheet.

Keywords: condenser, domestic refrigerator, heat transfer, mathematical model

Procedia PDF Downloads 441
4048 Drying and Transport Processes in Distributed Hydrological Modelling Based on Finite Volume Schemes (Iber Model)

Authors: Carlos Caro, Ernest Bladé, Pedro Acosta, Camilo Lesmes

Abstract:

The drying-wet process is one of the topics to be more careful in distributed hydrological modeling using finite volume schemes as a means of solving the equations of Saint Venant. In a hydrologic and hydraulic computer model, surface flow phenomena depend mainly on the different flow accumulation and subsequent runoff generation. These accumulations are generated by routing, cell by cell, from the heights of water, which begin to appear due to the rain at each instant of time. Determine when it is considered a dry cell and when considered wet to include in the full calculation is an issue that directly affects the quantification of direct runoff or generation of flow at the end of a zone of contribution by accumulations flow generated from cells or finite volume.

Keywords: hydrology, transport processes, hydrological modelling, finite volume schemes

Procedia PDF Downloads 375
4047 Colonization of Embrionic Gonads of Nile Tilapia by Giant Gourami Testicular Germ Cells

Authors: Irma Andriani, Ita Djuwita, Komar Sumantadinata, Alimuddin

Abstract:

The recent study has been conducted to develop testicular germ cell transplantation as a tool for preservation and propagation of male germ-plasm from endangered fish species, as well as to produce surrogate broodstock of commercially valuable fish. Giant gourami testis had been used as a model for donor and Nile tilapia larvae as recipient. We developed testicular cell xenotransplantation by optimizing the timing of intraperitoneal cell transplantation to recipient larvae aged 1, 3, 5 and 7 days post hatching (dph). Freshly isolated testis of giant gourami weighing 600–800 g were minced in dissociation medium and then incubated for 3 hours in room temperature to collect monodisperce cell suspension. Donor cells labeled with PKH 26 were transplanted into the peritoneal cavity of Nile tilapia larvae using glass micropipettes. Parameters observed were survival rate of Nile tilapia larvae at 24 hours post transplantation (pt) and colonization efficiency of donor cells at 2 and 3 months pt. The incorporated donor cells were observed under fluorescent microscope. The result showed that the lowest survival rate at 24 hours pt was 1 dph larvae (82.74±6.76%) and the highest survival rate were 3 and 5 dph larvae (95.00±5.00% and 95.00±2.50%, respectively). The highest colonization efficiency was on 3 dph larvae (61.1±34.71%) and the lowest colonization efficiency was on 7 dph larvae (19.43±17.33%). In conclusion, 3 dph Nile tilapia larvae was the best recipient for giant gourami testicular germ cells xenotransplantation.

Keywords: xenotransplantation, testicular germ cell, giant gourami, Nile tilapia, colonization efficiency

Procedia PDF Downloads 569