Search results for: adipose tissue derived stromal/stem cell

Authors: Ipsita Biswas, Arnab Sarkar, Ashikur Rahaman, Gopeswar Mukherjee, Subhrangsu Chatterjee, Shamee Bhattacharjee, Deba Prasad Mandal

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One of the major reasons for the high mortality rate of lung cancer is the substantial delays in disease detection at late metastatic stages. It is of utmost importance to understand the detailed molecular signaling and detect the molecular markers that can be used for the early diagnosis of cancer. Several studies explored the emerging roles of long noncoding RNAs (lncRNAs) in various cancers as well as lung cancer. A long non-coding RNA LINC00273 was recently discovered to promote cancer cell migration and invasion, and its positive correlation with the pathological stages of metastasis may prove it to be a potential target for inhibiting cancer cell metastasis. Comparing real-time expression of LINC00273 in various human clinical cancer tissue samples with normal tissue samples revealed significantly higher expression in cancer tissues. This long intergenic noncoding RNA was found to be highly expressed in human liver tumor-initiating cells, human gastric adenocarcinoma AGS cell line, as well as human non-small cell lung cancer A549 cell line. SiRNA and shRNA-induced knockdown of LINC00273 in both in vitro and in vivo nude mice significantly subsided AGS and A549 cancer cell migration and invasion. LINC00273 knockdown also reduced TGF-β induced SNAIL, SLUG, VIMENTIN, ZEB1 expression, and metastasis in A549 cells. Plenty of reports have suggested the role of microRNAs of the miR200 family in reversing epithelial to mesenchymal transition (EMT) by inhibiting ZEB transcription factors. In this study, hsa-miR-200a-3p was predicted via IntaRNA-Freiburg RNA tools to be a potential target of LINC00273 with a negative free binding energy of −8.793 kcal/mol, and this interaction was verified as a confirmed target of LINC00273 by RNA pulldown, real-time PCR and luciferase assay. Mechanistically, LINC00273 accelerated TGF-β induced EMT by sponging hsa-miR-200a-3p which in turn liberated ZEB1 and promoted prometastatic functions in A549 cells in vitro as verified by real-time PCR and western blotting. The similar expression patterns of these EMT regulatory pathway molecules, viz. LINC00273, hsa-miR-200a-3p, ZEB1 and TGF-β, were also detected in various clinical samples like breast cancer tissues, oral cancer tissues, lung cancer tissues, etc. Overall, this LINC00273 mediated EMT regulatory signaling can serve as a potential therapeutic target for the prevention of lung cancer metastasis.

Keywords: epithelial to mesenchymal transition, long noncoding RNA, microRNA, non-small-cell lung carcinoma

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Authors: Z. Karahaliloğlu, E. Yalçın, M. Demirbilek, E.B. Denkbaş

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Critical-sized bone defects caused by trauma, bone diseases, prosthetic implant revision or tumor excision cannot be repaired by physiological regenerative processes. Current orthopedic applications for critical-sized bone defects are to use autologous bone grafts, bone allografts, or synthetic graft materials. However, these strategies are unable to solve completely the problem, and motivate the development of novel effective biological scaffolds for tissue engineering applications and regenerative medicine applications. In particular, scaffolds combined with a variety of bio-agents as fundamental tools emerge to provide the regeneration of damaged bone tissues due to their ability to promote cell growth and function. In this study, a magnetic silk fibroin (SF) hydrogel scaffold was prepared by electrogelation process of the concentrated Bombxy mori silk fibroin (8 %wt) aqueous solution. For enhancement of osteoblast-like cells (SaOS-2) growth and adhesion, basal fibroblast growth factor (bFGF) were conjugated physically to the HSA-coated magnetic nanoparticles (Fe3O4) and magnetic SF e-gel scaffolds were prepared by incorporation of Fe3O4, HSA (human serum albumin)=Fe3O4 and HSA=Fe3O4-bFGF nanoparticles. HSA=Fe3O4, HSA=Fe3O4-bFGF loaded and bare SF e-gels scaffolds were characterized using scanning electron microscopy (SEM.) For cell studies, human osteoblast-like cell line (SaOS-2) was used and an MTT assay was used to assess the cytotoxicity of magnetic silk fibroin e-gel scaffolds and cell density on these surfaces. For the evaluation osteogenic activation, ALP (alkaline phosphatase), the amount of mineralized calcium, total protein and collagen were studied. Fe3O4 nanoparticles were successfully synthesized and bFGF was conjugated to HSA=Fe3O4 nanoparticles with %97.5 of binding yield which has a particle size of 71.52±2.3 nm. Electron microscopy images of the prepared HSA and bFGF incorporated SF e-gel scaffolds showed a 3D porous morphology. In terms of water uptake results, bFGF conjugated HSA=Fe3O4 nanoparticles has the best water absorbability behavior among all groups. In the in-vitro cell culture studies realized using SaOS-2 cell line, the coating of Fe3O4 nanoparticles surface with a protein enhance the cell viability and HSA coating and bFGF conjugation, the both have an inductive effect in the cell proliferation. One of the markers of bone formation and osteoblast differentiation, according to the ALP activity and total protein results, HSA=Fe3O4-bFGF loaded SF e-gels had significantly enhanced ALP activity. Osteoblast cultured HSA=Fe3O4-bFGF loaded SF e-gels deposited more calcium compared with SF e-gel. The proposed magnetic scaffolds seem to be promising for bone tissue regeneration and used in future work for various applications.

Keywords: basic fibroblast growth factor (bFGF), e-gel, iron oxide nanoparticles, silk fibroin

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Authors: R. P. Hewawasam, M. H. A. D. de Silva, M. A. G. Iresha

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In recent years childhood obesity has increased to pan-epidemic proportions along with a concomitant increase in obesity-associated morbidity. Birth weight is an important determinant of later adult health, with neonates at both ends of the birth weight spectrum at risk of future health complications. Consequently, infants who are born large for gestational age (LGA) are more likely to be obese in childhood and adolescence and are at risk of cardiovascular and metabolic complications later in life. Adipose tissue plays a role in linking events in fetal growth to the subsequent development of adult diseases. In addition to its role as a storage depot for fat, adipose tissue produces and secrets a number of hormones of importance in modulating metabolism and energy homeostasis. Cord blood leptin level has been positively correlated with fetal adiposity at birth. It is established that Asians have lower skeletal muscle mass, low bone mineral content and excess body fat for a given body mass index indicating a genetic predisposition in the occurrence of obesity. To our knowledge, studies have never been conducted in Sri Lanka to determine the relationship between adipocytokine profile in cord blood and anthropometric parameters in newborns. Thus, the objective of this study is to establish the above relationship for the Sri Lankan population to implement awareness programs to minimize childhood obesity in the future. Umbilical cord blood was collected from 90 newborns (Male 40, Female 50; gestational age 35-42 weeks) after double clamping the umbilical cord before separation of the placenta and the concentration of leptin was measured by ELISA technique. Anthropometric parameters of the newborn such as birth weight, length, ponderal index, occipital frontal, chest, hip and calf circumferences were measured. Pearson’s correlation was used to assess the relationship between leptin and anthropometric parameters while the Mann-Whitney U test was used to assess the differences in cord blood leptin levels between small for gestational age (SGA), appropriate for gestational age (AGA) and LGA infants. There was a significant difference (P < 0.05) between the cord blood leptin concentrations of LGA infants (12.67 ng/mL ± 2.34) and AGA infants (7.10 ng/mL ± 0.90). However, a significant difference was not observed between leptin levels of SGA infants (8.86 ng/mL ± 0.70) and AGA infants. In both male and female neonates, umbilical leptin levels showed significant positive correlations (P < 0.05) with birth weight of the newborn, pre-pregnancy maternal weight and pre pregnancy BMI between the infants of large and appropriate for gestational ages. Increased concentrations of leptin levels in the cord blood of large for gestational age infants suggest that they may be involved in regulating fetal growth. Leptin concentration of Sri Lankan population was not significantly deviated from published data of Asian populations. Fetal leptin may be an important predictor of neonatal adiposity; however, interventional studies are required to assess its impact on the possible risk of childhood obesity.

Keywords: appropriate for gestational age, childhood obesity, leptin, anthropometry

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Authors: Scott Nielsen, Luca Longanesi, Chris Chuck

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Palm oil is obtained from the flesh and kernel of the fruit of oil palms and is the most productive and inexpensive oil crop. The global demand for palm oil is approximately 75 million metric tonnes, a 29% increase in global production of palm oil since 2016. This expansion of oil palm cultivation has resulted in mass deforestation, vast biodiversity destruction and increasing net greenhouse gas emissions. One possible alternative is to produce a saturated oil, similar to palm, from microbes such as oleaginous yeast. The yeasts can be cultured on sugars derived from second-generation sources and do not compete with tropical forests for land. One highly promising oleaginous yeast for this application is Metschnikowia pulcherrima. However, recent techno-economic modeling has shown that cell lysis and standard lipid extraction are major contributors to the cost of the oil. Typical cell disruption techniques to extract either single cell oils or proteins have been based around bead-beating, homogenization and acid lysis. However, these can have a detrimental effect on lipid quality and are energy-intensive. In this study, a vortex separator, which produces high sheer with minimal energy input, was investigated as a potential low energy method of lysing cells. This was compared to four more traditional methods (thermal lysis, acid lysis, alkaline lysis, and osmotic lysis). For each method, the yeast loading was also examined at 1 g/L, 10 g/L and 100 g/L. The quality of the cell disruption was measured by optical cell density, cell counting and the particle size distribution profile comparison over a 2-hour period. This study demonstrates that the vortex separator is highly effective at lysing the cells and could potentially be used as a simple apparatus for lipid recovery in an oleaginous yeast process. The further development of this technology could potentially reduce the overall cost of microbial lipids in the future.

Keywords: palm oil substitute, metschnikowia pulcherrima, cell disruption, cell lysis

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Authors: Piamsiri Sawaisorn, Tienrat Tangchaikeeree, Waraporn Chan-On, Chaniya Leepiyasakulchai, Rachanee Udomsangpetch, Suradej Hongeng, Kulachart Jangpatarapongsa

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Human Vγ9Vδ2 T lymphocytes are regarded as promising effector cells for cancer immunotherapy since they have the ability to eliminate several tumor cells through non-peptide antigen recognition and non-major histocompatibility complex (MHC) restriction. An issue of recent interest is the capability to activate γδ T cells by use of a group of drugs, such as pamidronate, that cause accumulation of phosphoantigen which is recognized by γδ T cell receptors. Moreover, their antigen presenting cell-like phenotype and function have been confirmed in many clinical trials. In this study, Vγ9Vδ2 T cells derived from normal peripheral blood mononuclear cells were activated with pamidronate and the expanded Vγ9Vδ2 T cells can recognize and kill chronic myeloid leukemia (CML) cells treated with pamidronate through their cytotoxic activity. To support the strong role played by Vγ9Vδ2 T cells against cancer, we provide the evidence that Vγ9Vδ2 T cells activated with CML cell lysate antigen can efficiently express antigen presenting cell (APC) phenotype and function. In conclusion, pamidronate can be used in intentional activation of human Vγ9Vδ2 T cells and can increase the susceptibility of CML cells to cytotoxicity of Vγ9Vδ2 T cells. The activated Vγ9Vδ2 T cells by cancer cells lysate can show their APC characteristics, and so greatly increase the interest in exploring their therapeutic potential in hematologic malignancy.

Keywords: γδ T lymphocytes, antigen-presenting cells, chronic myeloid leukemia, cancer, immunotherapy

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Authors: Jefunnie Matahum, Yu-Chi Kuo, Chao-Ming Su, Tzong-Rong Ger

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Magnetic cell labeling is of great importance in various applications in biomedical fields such as cell separation and cell sorting. Since analytical methods for quantification of cell uptake of magnetic nanoparticles (MNPs) are already well established, image analysis on single cell level still needs more characterization. This study reports an alternative non-destructive quantification methods of single-cell uptake of positively charged MNPs. Magnetophoresis experiments were performed to calculate the number of MNPs in a single cell. Mobility of magnetic cells and the area of intracellular MNP stained by Prussian blue were quantified by image processing software. ICP-MS experiments were also performed to confirm the internalization of MNPs to cells. Initial results showed that the magnetic cells incubated at 100 µg and 50 µg MNPs/mL concentration move at 18.3 and 16.7 µm/sec, respectively. There is also an increasing trend in the number and area of intracellular MNP with increasing concentration. These results could be useful in assessing the nanoparticle uptake in a single cell level.

Keywords: magnetic nanoparticles, single cell, magnetophoresis, image analysis

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Authors: M. A. Islam, P. J. Hazell, J. P. Escobedo, M. Saadatfar

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Quasistatic compression and micro structural characterization of closed cell aluminium foams of different pore size and cell distributions has been carried out. Metallic foams have good potential for lightweight structures for impact and blast mitigation and therefore it is important to find out the optimized foam structure (i.e. cell size, shape, relative density, and distribution) to maximize energy absorption. In this paper, we present results for two different aluminium metal foams of density 0.5 g/cc and 0.7 g/cc respectively that have been tested in quasi-static compression. The influence of cell geometry and cell topology on quasistatic compression behavior has been investigated using computed tomography (micro-CT) analysis. The compression behavior and micro structural characterization will be presented.

Keywords: metal foams, micro-CT, cell topology, quasistatic compression

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Authors: Ji Su Kim, Bo Ram Choi, Ju Yeon Cho, Hyukjin Lee

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Epidermal growth factor receptor (EGFR) 19 deletion mutation gene is over-expressed in carcinoma patient. EGFR 19 deletion mutation is known as typical biomarker of non-small cell lung cancer (NSCLC), which one section in the coding exon 19 of EGFR is deleted. Therefore, there have been many attempts over the years to detect EGFR 19 deletion mutation for replacing conventional diagnostic method such as PCR and tissue biopsy. We developed a simple and facile detection platform based on Rolling Circle Amplification (RCA), which provides highly amplified products in isothermal amplification of the ligated DNA template. Limit of detection (~50 nM) and a faster detection time (~30 min) could be achieved by introducing RCA.

Keywords: EGFR19, cancer, diagnosis, rolling circle amplification (RCA), hydrogel

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Authors: Ashima Sharma

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Human Serum Albumin (HSA)- one of the most demanded therapeutic proteins with immense biotechnological applications- is a large multidomain protein containing 17 disulfide bonds. The current source of HSA is human blood plasma which is a limited and unsafe source. Thus, there exists an indispensable need to promote non-animal derived recombinant HSA (rHSA) production. Escherichia coli is one of the most convenient hosts which had contributed to the production of more than 30% of the FDA approved recombinant pharmaceuticals. It grows rapidly and reaches high cell density using inexpensive and simple substrates. E. coli derived recombinant products have more economic potential as fermentation processes are cheaper compared to the other expression hosts. The major bottleneck in exploiting E. coli as a host for a disulfide-rich multidomain protein is the formation of aggregates of overexpressed protein. The majority of the expressed HSA forms inclusion bodies (more than 90% of the total expressed rHSA) in the E. coli cytosol. Recovery of functional rHSA from inclusion bodies is not preferred because it is difficult to obtain a large multidomain disulfide bond rich protein like rHSA in its functional native form. Purification is tedious, time-consuming, laborious and expensive. Because of such limitations, the E. coli host system was neglected for rHSA production for the past few decades despite its numerous advantages. In the present work, we have exploited the capabilities of E. coli as a host for the enhanced functional production of rHSA (~60% of the total expressed rHSA in the soluble fraction). Parameters like intracellular environment, temperature, induction type, duration of induction, cell lysis conditions etc. which play an important role in enhancing the level of production of the desired protein in its native form in vivo have been optimized. We have studied the effect of assistance of different types of exogenously employed chaperone systems on the functional expression of rHSA in the E. coli host system. Different aspects of cell growth parameters during the production of rHSA in presence and absence of molecular chaperones in E. coli have also been studied. Upon overcoming the difficulties to produce functional rHSA in E. coli, it has been possible to produce significant levels of functional protein through engineering the biological system of protein folding in the cell, the E. coli-derived rHSA has been purified to homogeneity. Its detailed physicochemical characterization has been performed by monitoring its conformational properties, secondary and tertiary structure elements, surface properties, ligand binding properties, stability issues etc. These parameters of the recombinant protein have been compared with the naturally occurring protein from the human source. The outcome of the comparison reveals that the recombinant protein resembles exactly the same as the natural one. Hence, we propose that the E. coli-derived rHSA is an ideal biosimilar for human blood plasma-derived serum albumin. Therefore, in the present study, we have introduced and promoted the E. coli- derived rHSA as an alternative to the preparation from a human source, pHSA.

Keywords: recombinant human serum albumin, Escherichia coli, biosimilar, chaperone assisted protein folding

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Authors: Mohammad Syahirin Aisha, Khairul Imran Sainan

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The PEM fuel cell is a device that generate electric by electrochemical reaction between hydrogen fuel and oxygen in the fuel cell stack. PEM fuel cell consists of an anode (hydrogen supply), a cathode (oxygen supply) and an electrolyte that allow charges move between the two positions of the fuel cell. The only product being developed after the reaction is water (H2O) and heat as the waste which does not emit greenhouse gasses. The performance of fuel cell affected by numerous parameters. This study is restricted to cathode parameters that affect fuel cell performance. At the anode side, the reactant is not going through any changes. Experiments with variation in air velocity (3m/s, 6m/s and 9m/s), temperature (10oC, 20oC, 35oC) and relative humidity (50%, 60%, and 70%) have been carried out. The experiments results are presented in the form of fuel cell stack power output over time, which demonstrate the impacts of the various air condition on the execution of the PEM fuel cell. In this study, the experimental analysis shows that with variation of air conditions, it gives different fuel cell performance behavior. The maximum power output of the experiment was measured at an ambient temperature of 25oC with relative humidity and 9m/s velocity of air.

Keywords: air-breathing PEM fuel cell, cathode side, performance, variation in air condition

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Authors: Hilal Turkoglu Sasmazel, Ozan Ozkan, Seda Surucu

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Tissue engineering is the field of treating defects caused by injuries, trauma or acute/chronic diseases by using artificial scaffolds that mimic the extracellular matrix (ECM), the natural biological support for the tissues and cells within the body. The main aspects of a successful artificial scaffold are (i) large surface area in order to provide multiple anchorage points for cells to attach, (ii) suitable porosity in order to achieve 3 dimensional growth of the cells within the scaffold as well as proper transport of nutrition, biosignals and waste and (iii) physical, chemical and biological compatibility of the material in order to obtain viability throughout the healing process. By hybrid scaffolds where two or more different materials were combined with advanced fabrication techniques into complex structures, it is possible to combine the advantages of individual materials into one single structure while eliminating the disadvantages of each. Adding this to the complex structure provided by advanced fabrication techniques enables obtaining the desired aspects of a successful artificial tissue scaffold. In this study, fibroblast compatibility of poly(ε-caprolactone) (PCL)/chitosan core-shell electrospun hybrid scaffolds with proper mechanical, chemical and physical properties successfully developed in our previous study was investigated. Standard 7-day cell culture was carried out with L929 fibroblast cell line. The viability of the cells cultured with the scaffolds was monitored with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) viability assay for every 48 h starting with 24 h after the initial seeding. In this assay, blank commercial tissue culture polystyrene (TCPS) Petri dishes, single electrospun PCL and single electrospun chitosan mats were used as control in order to compare and contrast the performance of the hybrid scaffolds. The adhesion, proliferation, spread and growth of the cells on/within the scaffolds were observed visually on the 3rd and the 7th days of the culture period with confocal laser scanning microscopy (CSLM) and scanning electron microscopy (SEM). The viability assay showed that the hybrid scaffolds caused no toxicity for fibroblast cells and provided a steady increase in cell viability, effectively doubling the cell density for every 48 h for the course of 7 days, as compared to TCPS, single electrospun PCL or chitosan mats. The cell viability on the hybrid scaffold was ~2 fold better compared to TCPS because of its 3D ECM-like structure compared to 2D flat surface of commercially cell compatible TCPS, and the performance was ~2 fold and ~10 fold better compared to single PCL and single chitosan mats, respectively, even though both fabricated similarly with electrospinning as non-woven fibrous structures, because single PCL and chitosan mats were either too hydrophobic or too hydrophilic to maintain cell attachment points. The viability results were verified with visual images obtained with CSLM and SEM, in which cells found to achieve characteristic spindle-like fibroblast shape and spread on the surface as well within the pores successfully at high densities.

Keywords: chitosan, core-shell, fibroblast, electrospinning, PCL

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

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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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Authors: Mehdi Abbasi, Shadan Navid, Mohammad Pourahmadi, M. Majidi Zolbin

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We have recently reported that melatonin as antioxidant enhances the efficacy of colonization of spermatogonial stem cells (SSCs). Melatonin as an antioxidant plays a vital role in the development of SSCs in vitro. This study aimed to investigate evaluation of sertoli cells and melatonin simultaneously on SSC proliferation following transplantation to testis of adult mouse busulfan-treated azoospermia model. SSCs and sertoli cells were isolated from the testes of three to six-day old male mice.To determine the purity, Flow cytometry technique using PLZF antibody were evaluated. Isolated testicular cells were cultured in αMEM medium in the absence (control group) or presence (experimental group) of sertoli cells and melatonin extract for 2 weeks. We then transplanted SSCs by injection into the azoospermia mice model. Higher viability, proliferation, and Id4, Plzf, expression were observed in the presence of simultaneous sertoli cells and melatonin in vitro. Moreover, immunocytochemistry results showed higher Oct4 expression in this group. Eight weeks after transplantation, injected cells were localized at the base of seminiferous tubules in the recipient testes. The number of spermatogonia and the weight of testis were higher in the experimental group relative to control group. The results of our study suggest that this new protocol can increase the transplantation of these cells can be useful in the treatment of male infertility.

Keywords: colonization, melatonin, spermatogonial stem cell, transplantation

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Authors: Chuan Yin

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Cancer stem cells (CSCs) represent a subpopulation of cancer cells that possess the characteristics associated with normal stem cells. CD133 is a phenotype of melanoma CSCs responsible for melanoma metastasis and drug resistance. Although adriamycin (ADR) is commonly used drug in melanoma therapy, but it is ineffective in the treatment of melanoma CSCs. In this study, we constructed CD133 antibody conjugated ADR immunoliposomes (ADR-Lip-CD133) to target CD133+ melanoma CSCs. The results showed that the immunoliposomes possessed a small particle size (~150 nm), high drug encapsulation efficiency (~90%). After 72 hr treatment on the WM266-4 melanoma tumorspheres, the IC50 values of the drug formulated in ADR-Lip-CD133, ADR-Lip (ADR liposomes) and ADR are found to be 24.42, 57.13 and 59.98 ng/ml respectively, suggesting that ADR-Lip-CD133 was more effective than ADR-Lip and ADR. Significantly, ADR-Lip-CD133 could almost completely abolish the tumorigenic ability of WM266-4 tumorspheres in vivo, and showed the best therapeutic effect in WM266-4 melanoma xenograft mice. It is noteworthy that ADR-Lip-CD133 could selectively kill CD133+ melanoma CSCs of WM266-4 cells both in vitro and in vivo. ADR-Lip-CD133 represent a potential approach in targeting and killing CD133+ melanoma CSCs.

Keywords: cancer stem cells, melanoma, immunoliposomes, CD133

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Authors: Khaled A. shady, Fagr B. Bazeed, Nashwa K. Abousamra, Ihab H. Elberawe, Ashraf E. shaalan, Mohamed A. Sobh

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Physical activity activates a variety of adult stem cells which might be released into the circulation or might be activated in their organ-resident state. A variety of stimuli such as metabolic, mechanical, and hormonal stimuli might by responsible for the mobilization. This study was done to know the changes in hematopoietic stem cells and endothelial progenitor in athletes in the 24 hours following 30 min of aerobic exercise. Methods: Ten healthy male's athlete's (age 20.7± 0.61 y) performed moderate running with 30 min at 80% of velocity of The IAT. Blood samples taken pre-, and immediately, 30 min, 2h, 6h and 24h post-exercise were analyzed for hematopoietic stem cells (HSCs ), endothelial progenitor cells (EPCs(, vascular endothelial growth factor (VEGF), nitric oxide (NO), lactic acid (LA), and white blood cells . HSCs and EPCs were quantified by flow cytometry. Results: After 30min of aerobic exercise significant increases in HSCs, EPC, VEGF, NO, LA and WBCs (p ˂ 0.05). This increase will be at different rates according to the timing of taking blood sample and was in the maximum rate of increase after 30 min of aerobic exercise. HSCs, EPC, NO and WBCs were in the maximum rate of increase 2h post exercise. In addition, VEGF was in the maximum rate of increase immediately post exercise and LA concentration not affected after exercise. Conclusion: These data suggest that HSCs and EPCs increased after aerobic exercise due to increase of VEGF which play an important role in mobilization of stem cells and promotes NO increase which contributes to increase EPCs.

Keywords: physical activity, hematopoietic stem cells, mobilization, athletes

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Authors: Anne Bubriski

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Despite improvements in gender inequalities, women and girls continue to face glass ceilings, underrepresentation, and harmful stereotypes that can limit their aspirations and opportunities in STEM. While girls are taking similar high school math and science classes, boys are more likely to take physics and six times more likely to take an engineering course. The gap becomes even larger for minority or low-income girls. This gender gap is not due to biology; rather, it is due to cultural, social, and institutional forces. As girls get older, these forces often ‘teach’ them ‘STEM is more for boys’. The STEM gender gap widens in college, with only 20% of engineering degrees being awarded to women, and by the time women enter the workforce, they only occupy about 13% of engineering jobs. At the University of Central Florida, the Women’s and Gender Studies Program has developed a unique mentoring program to address these issues, Science Leadership and Mentoring (SLAM). What is unique about the approach of SLAM is that we look to address this problem through leadership and STEM. We look to help girls make connections between leadership and STEM—that young women can be leaders as scientists and that scientists are leaders making a change. This is particularly needed and relevant to our community because while there are mentoring programs to our knowledge, SLAM is one of the only, if not only, mentoring programs pairing college women and 7th-grade girls that includes a focus both on STEM and leadership in the United States. SLAM is a curriculum-based mentoring program pairing one 7th-grade girl with one UCF undergraduate STEM major. SLAM empowers young women to be assertive, brave, confident, independent, inquisitive and proud leaders in STEM. SLAM seeks to promote young women’s inspiration and excitement into STEM fields and careers while also building leadership abilities such as problem-solving, teamwork and cooperation, cultural identity and ethnic pride, advocacy for positive change, and goals for the future. SLAM serves about fifteen 7th-grade girls for the academic year and about 20 UCF students. SLAM holds weekly mentoring meetings lasting about 90 minutes, covering topics on leadership, STEM majors and careers, and STEM leadership. This past year, SLAM received a Community Action Grant from the American Association of University Women (AAUW) to run a sub-program, SLAM-Space. SLAM-Space focused on exposing SLAM participants to aerospace engineering and other space-related STEM fields, such as physics and astronomy, through guest speakers, workshops and field trips, including the Kenndy Space Center. The proposed paper presentation will present an overview of SLAM-Space and the data findings from pre and post-surveys, in-depth interviews and focus groups from the SLAM participants' experiences in the program.

Keywords: gender, leadership, STEM, empowerment

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Authors: Mustafa M. Donma, Orkide Donma

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Obesity is a low-grade inflammatory state. Childhood obesity is a multisystem disease, which is associated with a number of complications as well as potentially negative consequences. Gender is an important universal risk factor for many diseases. Hematological indices differ significantly by gender. This should be considered during the evaluation of obese children. The aim of this study is to detect hematologic indices that differ by gender in morbid obese (MO) children. A total of 134 MO children took part in this study. The parents filled an informed consent form and the approval from the Ethics Committee of Namik Kemal University was obtained. Subjects were divided into two groups based on their genders (64 females aged 10.2±3.1 years and 70 males aged 9.8±2.2 years; p ≥ 0.05). Waist-to-hip as well as head-to-neck ratios and body mass index (BMI) values were calculated. The children, whose WHO BMI-for age and sex percentile values were > 99 percentile, were defined as MO. Hematological parameters [haemoglobin, hematocrit, erythrocyte count, mean corpuscular volume, mean corpuscular haemoglobin, mean corpuscular haemoglobin concentration, red blood cell distribution width, leukocyte count, neutrophil %, lymphocyte %, monocyte %, eosinophil %, basophil %, platelet count, platelet distribution width, mean platelet volume] were determined by the automatic hematology analyzer. SPSS was used for statistical analyses. P ≤ 0.05 was the degree for statistical significance. The groups included children having mean±SD value of BMI as 26.9±3.4 kg/m² for males and 27.7±4.4 kg/m² for females (p ≥ 0.05). There was no significant difference between ages of females and males (p ≥ 0.05). Males had significantly increased waist-to-hip ratios (0.95±0.08 vs 0.91±0.08; p=0.005) and mean corpuscular hemoglobin concentration values (33.6±0.92 vs 33.1±0.83; p=0.001) compared to those of females. Significantly elevated neutrophil (4.69±1.59 vs 4.02±1.42; p=0.011) and neutrophil-to-lymphocyte ratios (1.70±0.71 vs 1.39±0.48; p=0.004) were detected in females. There was no statistically significant difference between groups in terms of C-reactive protein values (p ≥ 0.05). Adipose tissue plays important roles during the development of obesity and associated diseases such as metabolic syndrom and cardiovascular diseases (CVDs). These diseases may cause changes in complete blood cell count parameters. These alterations are even more important during childhood. Significant gender effects on the changes of neutrophils, one of the white blood cell subsets, were observed. The findings of the study demonstrate the importance of considering gender in clinical studies. The males and females may have distinct leukocyte-trafficking profiles in inflammation. Female children had more circulating neutrophils, which may be the indicator of an increased risk of CVDs, than male children within this age range during the late stage of obesity. In recent years, females represent about half of deaths from CVDs; therefore, our findings may be the indicator of the increasing tendency of this risk in females starting from childhood.

Keywords: children, gender, morbid obesity, neutrophil-to-lymphocyte ratio

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Authors: Ashwani Kumar

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Use of NIR light for imaging the biological tissue and to quantify its optical properties is a good choice over other invasive methods. Optical tomography involves two steps. One is the forward problem and the other is the reconstruction problem. The forward problem consists of finding the measurements of transmitted light through the tissue from source to detector, given the spatial distribution of absorption and scattering properties. The second step is the reconstruction problem. In X-ray tomography, there is standard method for reconstruction called filtered back projection method or the algebraic reconstruction methods. But this method cannot be applied as such, in optical tomography due to highly scattering nature of biological tissue. A hybrid algorithm for reconstruction has been implemented in this work which takes into account the highly scattered path taken by photons while back projecting the forward data obtained during Monte Carlo simulation. The reconstructed image suffers from blurring due to point spread function.

Keywords: NIR light, tissue, blurring, Monte Carlo simulation

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Authors: N. Chetty, K. Singh

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In this work, the relationship between the melanin content in a tissue and subsequent absorption of light through that tissue was determined using a digital camera. This technique proved to be simple, cost effective, efficient and reliable. Tissue phantom samples were created using milk and soy sauce to simulate the optical properties of melanin content in human tissue. Increasing the concentration of soy sauce in the milk correlated to an increase in melanin content of an individual. Two methods were employed to measure the light transmitted through the sample. The first was direct measurement of the transmitted intensity using a conventional lux meter. The second method involved correctly calibrating an ordinary digital camera and using image analysis software to calculate the transmitted intensity through the phantom. The results from these methods were then graphically compared to the theoretical relationship between the intensity of transmitted light and the concentration of absorbers in the sample. Conclusions were then drawn about the effectiveness and efficiency of these low cost methods.

Keywords: tissue phantoms, scattering coefficient, albedo, low-cost method

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Authors: Mpofu Vongai, Vhurumuku Elaosi

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Many African countries, such as Zimbabwe and South Africa, have curricula reform agendas that include incorporation of Indigenous Knowledge and Nature of Science (NOS) into school Science, Technology, Engineering and Mathematics (STEM) education. It is argued that at high school level, STEM learning, which incorporates understandings of indigenization science and NOS, has the potential to provide a strong foundation for a culturally embedded scientific knowledge essential for their advancement in Science and Technology. Globally, investment in STEM education is recognized as essential for economic development. For this reason, developing countries such as Zimbabwe and South Africa have been investing into training specialized teachers in natural sciences and technology. However, in many cases this training has been detached from the cultural realities and contexts of indigenous learners. For this reason, the STEM curricula reform has provided implementation challenges to teachers. An issue of major concern is the teachers’ pedagogical content knowledge (PCK), which is essential for effective implementation of these STEM curricula. Well-developed Teacher PCK include an understanding of both the nature of indigenous knowledge (NOIK) and of NOS. This paper reports the results of a study that investigated the development of 3 South African and 3 Zimbabwean in-service teachers’ abilities to integrate NOS and NOIK as part of their PCK. A participatory action research design was utilized. The main focus was on capturing, determining and developing teachers STEM knowledge for integrating NOIK and NOS in science classrooms. Their use of indigenous games was used to determine how their subject knowledge for STEM and pedagogical abilities could be developed. Qualitative data were gathered through the use dialogues between the researchers and the in-service teachers, as well as interviewing the participating teachers. Analysis of the data provides a methodological window through which in-service teachers’ PCK can be STEMITIZED and their abilities to integrate NOS and NOIK developed. Implications are raised for developing teachers’ STEM education in universities and teacher training colleges.

Keywords: indigenous knowledge, nature of science, pedagogical content knowledge, STEM education

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Authors: Mahfuzur Rahman

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Back-contact solar cells improve optical properties by moving all electrically conducting parts to the back of the cell. The cell's structure allows silicon solar cells to surpass the 25% efficiency barrier and interdigitated solar cells are now the most efficient. In this work, the fabrication of a light, efficient and temperature resistant interdigitated back contact (IBC) solar cell is investigated. This form of solar cell differs from a conventional solar cell in that the electrodes are located at the back of the cell, eliminating the need for grids on the top, allowing the full surface area of the cell to receive sunlight, resulting in increased efficiency. In this project, we will use SILVACO TCAD, an optoelectronic device simulator, to construct a very thin solar cell with dimensions of 100x250um in 2D Luminous. The influence of sunlight intensity and atmospheric temperature on solar cell output power is highly essential and it has been explored in this work. The cell's optimum performance with 150um bulk thickness provides 28.81% efficiency with an 87.68% fill factor rate making it very thin, flexible and resilient, providing diverse operational capabilities.

Keywords: interdigitated, shading, recombination loss, incident-plane, drift-diffusion, luminous, SILVACO

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Authors: Kevin Zhao, Norman J. Horing

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A methodology for cells sorting and circulating tumor cell detection and discrimination is presented in this paper. The technique is based on Dielectrophoresis and microfluidic device theory. Specifically, the sorting of the cells is realized by adjusting the relation among the sedimentation forces, the drag force provided by the fluid, and the Dielectrophortic force that is relevant to the bias voltage applied on the device. The relation leads to manipulation of the elevation of the cells of the same kind to a height by controlling the bias voltage. Once the cells have been lifted to a position next to the bottom of the cell collection channel, the buffer fluid flashes them into the cell collection channel. Repeated elevation of the cells leads to a complete sorting of the cells in the sample chamber. A proof-of-principle example is presented which verifies the feasibility of the methodology.

Keywords: cell sorter, CTC cell, detection and discrimination, dielectrophoresisords, simulation

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Authors: Cyril Deroy, Agata Rumianek, David R. Greaves, Peter R. Cook, Edmond J. Walsh

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Various microfluidic platforms have been developed in the past couple of decades offering experimental methods for the study of cell migration; however, their implementation in the laboratory has remained limited. Some reasons cited for the lack of uptake include the technical complexity of the devices, high failure rate associated with gas-bubbles, biocompatibility concerns with the use of polydimethylsiloxane (PDMS) and equipment/time/expertise requirements for operation and manufacture. As sample handling remains challenging due to the closed format of microfluidic devices, open microfluidic systems have been developed offering versatility and simplicity of use. Rather than confining fluids by solid walls, samples can be accessed directly over the open platform, by removing at least one of the solid boundaries, such as the cover. In this paper, a method for the fabrication of open fluid-walled microfluidic circuits for cell migration studies is introduced, where only materials commonly used by the life-science community are required; tissue culture dishes and cell media. The simplicity of the method, and ability to retrieve cells of interest are two key features of the method. Both passive and active flow-devices can be created in this way. To demonstrate the versatility of the method a cell migration assay is performed, which requires fabricating circuits for establishing chemical gradients, loading cells and incubating, creating chemical gradients, real time imaging of cell migration and finally retrieval of cells. The open architecture has high fidelity as it eliminates air bubble related failures and enables the precise control of gradients. The ability to fabricate custom microfluidic designs in minutes should make this method suitable for use in a wide range of cell migration studies.

Keywords: chemotaxis, fluid walls, gradient generation, open microfluidics

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Authors: Emily Schlebes, Christian Hundhausen, Jens W. Fischer

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The glycosaminoglycan hyaluronan (HA) is a major component of the extracellular matrix, typically produced by fibroblasts of the connective tissue but also by immune cells. Here, we investigated the capacity of human peripheral blood CD8 T cells from healthy donors to produce HA and to express HA receptors as well as HA degrading enzymes. Further, we evaluated the effect of pharmacological HA inhibition on CD8 T cell function. Using immunocytochemistry together with quantitative PCR analysis, we found that HA synthesis is rapidly induced upon antibody-induced T cell receptor (TCR) activation and almost exclusively mediated by HA synthase 3 (HAS3). TCR activation also resulted in the upregulation of HA receptors CD44, hyaluronan-mediated motility receptor (HMMR), and layilin (LAYN), although kinetics and strength of expression varied greatly between subjects. The HA-degrading enzymes HYAL1 and HYAL2 were detected at low levels and induced by cell activation in some individuals. Interestingly, expression of HAS3, HA receptors, and hyaluronidases were modulated by the proinflammatory cytokines IL-6 and IL-1bβ in most subjects. To assess the functional role of HA in CD8 T cells, we performed carboxyfluorescein succinimidyl ester (CFSE) based proliferation assays and cytokine analysis in the presence of the HA inhibitor 4- Methylumbelliferone (4-MU). Despite significant inter-individual variation with regard to the effective dose, 4-MU resulted in the inhibition of CD8 T cell proliferation and reduced release of TNF-α and IFN-γ. Collectively, these data demonstrate that human CD8 T cells respond to TCR stimulation with a synthesis of HA and expression of HA-related genes. They further suggest that HA inhibition may be helpful in interfering with pathogenic T cell activation in human disease.

Keywords: CD8 T cells, extracellular matrix, hyaluronan, hyaluronan synthase 3

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Authors: Kebret Kebede, Anthony Scinta

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Obesity is the most prevalent global problem that continues to rise at alarming rates both in the industrialized and developing countries. Adipose tissue is an endocrine tissue that secretes numerous chemical signals, hormones, lipids, cytokines and coagulation factors as well as prompting insulin resistance which is a primary contributor to Type II Diabetes- one of its most common adverse effects on health. Other hormones whose levels are linked to obesity and nutritional state are leptin, IGF-1, and adiponectin. Several studies indicate that obesity is the leading cause of high levels of cholesterol that leads to fatty liver disease, gallstones, hypertension, increased risk for cancer and degenerative joint disease that primarily affects the weight bearing joints of the lower extremities. The activation of inflammatory pathways promotes synovial pathology that results in accelerated degeneration of the joints. The study examines the prevalence of obesity in the US female population in comparison to that of the developing world and its emergence as a significant and potentially modifiable risk factor in degenerative disease of the hip and knee joints that has resulted in staggering healthcare cost. Studies have shown that as the prevalence of obesity rises, we continue to see a rise in degenerative joint disease. The percentage of arthritis cases linked directly to obesity has risen from 3 percent in 1971 to 18 percent in 2002. A person with obesity is around 60 percent more likely to develop arthritis than someone of normal body weight. In women, obesity is associated with increased mortality from breast, cervical, endometrial and ovarian cancer that may accompany debilitating joint diseases and restricted mobility.

Keywords: obesity, endocrine, degenerative, mortality, joint diseases, cancer, debilitating, mobility

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Authors: Naser Shagerdi Esmaeli, Mohsen Hamidpour

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Background and Objective: Hematopoietic stem cell transplantation (HSCT) has been widely used for treating oncological and hematological diseases. Although HSCT has helped to improve patient survival, the risk of developing an infection during hospitalization is an important cause of morbidity and mortality. This study aimed to analyze the infection profile during hospitalization and the associated risk factors among patients undergoing autologous HSCT at the University Hospital, Shahid Ghazi Tabatabaei Hospital, Tabriz, Iran. Subjects and Methods: This was a cross-sectional study on patients undergoing autologous HSCT at a public university hospital. Methods: Patients with febrile neutropenia between 2015 and 2018 were retrospectively evaluated regarding their infection profile and associated risk factors. This survey included: bacterial culture and blood culture on specific media. Results: Infection occurred in 57.2% of 56 patients with febrile neutropenia. The main source of infection was the central venous catheter (25.9%). Infection was chiefly due to Gram-positive bacteria, although Gram-negative-related infections were more severe and caused a higher death rate. Sex, age, skin color, nutritional status, and underlying disease were not associated with the development of infection. Patients with severe mucositis (Grades III and IV) had a higher infection rate (P < 0.001). Patients who developed pulmonary complications during hospitalization had higher infection rates (P = 0.002). Infection was the main cause of death (57.1%) in the study sample. Conclusion: Strategies aimed at reducing infection-related mortality rates among patients undergoing autologous HSCT are necessary.

Keywords: hematopoietic stem cell, autologous bone marrow transplantation, infection profile, tabriz, Iran

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Authors: Jong-Bae Lee, Seongsoo Lee

Abstract:

This paper proposes an adaptive discharge time control method to balance cell voltages in alternating battery cell discharging method. In the alternating battery cell discharging method, battery cells are periodically discharged in turn. Recovery effect increases battery output voltage while the given battery cell rests without discharging, thus battery operation time of target system increases. However, voltage mismatch between cells leads two problems. First, voltage difference between cells induces inter-cell current with wasted power. Second, it degrades battery operation time, since system stops when any cell reaches to the minimum system operation voltage. To solve this problem, the proposed method adaptively controls cell discharge time to equalize both cell voltages. In the proposed method, battery operation time increases about 19%, while alternating battery cell discharging method shows about 7% improvement.

Keywords: battery, recovery effect, low-power, alternating battery cell discharging, adaptive discharge time control

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Authors: Obinna K. Ihesiulor, Krishna Shankar, Paul Smith, Alan Fien

Abstract:

Excessive fretting wear at the taper-trunnion junction (trunnionosis) apparently contributes to the high failure rates of hip implants. Implant wear and corrosion lead to the release of metal particulate debris and subsequent release of metal ions at the taper-trunnion surface. This results in a type of metal poisoning referred to as metallosis. The consequences of metal poisoning include; osteolysis (bone loss), osteoarthritis (pain), aseptic loosening of the prosthesis and revision surgery. Follow up after revision surgery, metal debris particles are commonly found in numerous locations. Background: A stable connection between the femoral ball head (taper) and stem (trunnion) is necessary to prevent relative motions and corrosion at the taper junction. Hence, the importance of component assembly cannot be over-emphasized. Therefore, the aim of this study is to determine the influence of head-stem junction assembly by press fitting and the subsequent disengagement/disassembly on the connection strength between the taper ball head and stem. Methods: CoCr femoral heads were assembled with High stainless hydrogen steel stem (trunnion) by Push-in i.e. press fit; and disengaged by Pull-out test. The strength and stability of the two connections were evaluated by measuring the head pull-out forces according to ISO 7206-10 standards. Findings: The head-stem junction strength linearly increases with assembly forces.

Keywords: wear, modular hip prosthesis, taper head-stem, force assembly and disassembly

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Authors: Christoph Mayer, Dominik Holzmann

Abstract:

This paper presents the development of a compact, portable and easy to handle solar cell characterization device. The presented device reduces the effort and cost of single solar cell characterization to a minimum. It enables realistic characterization of cells under sunlight within minutes. In the field of photovoltaic research the common way to characterize a single solar cell or a module is, to measure the current voltage curve. With this characteristic the performance and the degradation rate can be defined which are important for the consumer or developer. The paper consists of the system design description, a summary of the measurement results and an outline for further developments.

Keywords: solar cell, photovoltaics, PV, characterization

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Authors: Gürol Önal, Kevser Dinçer, Salih Yayla

Abstract:

In this study, performance of proton exchange membrane PEM fuel cell was experimentally investigated. Coating on the anode side of the PEM fuel cell was accomplished with the spin method by using YSZ+SDC. A solution having 0,1 gr YttriaStabilized Zirconia (YSZ) + 0,1 Samarium-Doped Ceria (SDC) + 10 mL methanol was prepared. This solution was taken out and filled into a micro-pipette. Then the anode side of PEM fuel cell was coated with YSZ+ SDC by using spin method. In the experimental study, current, voltage and power performances before and after coating were recorded and then compared to each other. It was found that the efficiency of PEM fuel cell increases after the coating with YSZ+SDC.

Keywords: fuel cell, Polymer Electrolyte Membrane (PEM), membrane, spin method

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