Search results for: germinal center cells

Authors: Duygu Yüksel Deniz, Memet Vezir Kahraman, Serap Erdem Kuruca, Mediha Süleymanoğlu

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Our first aim was to synthesize Hydroxy Apatite (HAP) and then modify its surface by adding 4-Vinylbenzene boronic acid (4-VBBA). The characterization was done by FT-IR. By adding Polyvinyl alcohol (PVA) to 4- VBBA-HAP, we obtained a suitable electrospinning solution. PVA solution which was also modified by using alkoxy silanes, in order to prevent the scaffolds from being damaged by aqueous cell medium, was added. Keratin was dissolved and then added into the electrospinning solution. Keratin containing 4-VBBA- HAP/PVA composite was used to fabricate nanofiber scaffolds with the simultaneous UV-reactive electrospinning technique. The structural characterization was done by FT-IR. Thermal gravimetric analysis was also performed by using TGA. The morphological characterization was determined by SEM analyses. Our second aim was to create a scaffold where cells could grow. With this purpose, suitable nanofibers were choosen according to their SEM analysis. Keratin containing nanofibers were seeded with 3T3, ECV and SAOS cells and their cytotoxicity and cell proliferation were investigated by using MTT assay. After cell culturing process morphological characterization was determined by SEM analyses. These scaffolds were designed to be nontoxic biomaterials. Here, a comparision was made between keratin containing 3T3, ECV and SAOS seeded nanofiber scaffolds and the results were presented and discussed.

Keywords: cell culture, keratin, nanofibers, UV-reactive electrospinning

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Authors: Mohammad Afzal Khan, Fatimah Alanazi, Hala Abdalrahman Ahmed, Talal Shamma, Kilian Kelly, Mohammed A. Hammad, Abdullah O. Alawad, Abdullah Mohammed Assiri, Dieter Clemens Broering

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Lung transplantation is a life-saving surgical replacement of diseased lungs in patients with end-stage respiratory malfunctions. Despite the remarkable short-term recovery, long-term lung survival continues to face several significant challenges, including chronic rejection and severe toxic side-effects due to global immunosuppression. Stem cell-based immunotherapy has been recognized as a crucial immunoregulatory regimen in various preclinical and clinical studies. Despite initial therapeutic outcomes, conventional stem cells face key limitations. The Cymerus™ manufacturing facilitates the production of a virtually limitless supply of consistent human induced pluripotent stem cell (iPSC)-derived mesenchymal stem cells, which could play a key role in selective immunosuppression and graft repair during rejection. Here, we demonstrated the impact of iPSC-derived human MSCs on the development of immune-tolerance and long-term graft survival in mouse orthotopic airway allografts. BALB/c→C57BL/6 allografts were reconstituted with iPSC-derived MSCs (2 million/transplant/ at d0), and allografts were examined for regulatory T cells (Tregs), oxygenation, microvascular blood flow, airway epithelium and collagen deposition during rejection. We demonstrated that iPSC-derived MSC treatment leads to significant increase in tissue expression of hTSG-6 protein, followed by an upregulation of mouse Tregs and IL-5, IL-10, IL-15 cytokines, which augments graft microvascular blood flow and oxygenation, and thereby maintained a healthy airway epithelium and prevented the subepithelial deposition of collagen at d90 post-transplantation. Collectively, these data confirmed that iPSC-derived MSC-mediated immunosuppression has potential to establish immune-tolerance and rescue allograft from sustained hypoxic/ischemic phase and subsequently limits long-term airway epithelial injury and collagen progression, which therapeutically warrant a study of Cymerus iPSC-derived MSCs as a potential management option for immunosuppression in transplant recipients.

Keywords: stem cell therapy, immunotolerance, regulatory T cells, hypoxia and ischemia, microvasculature

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Authors: Mashitoh Abd Rahman, Najihah Mohd Hashim, Faiqah Ramli, Syam Mohan, Noraziah Nordin, Hamed Karimian, Hapipah Mohd Ali

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Ovarian cancer is the most lethal gynecological malignancies. HME5, a prenylflavanoid has been isolated from local medicinal plant. This compound has been reported to possess a broad spectrum of biological activities including anticancer property. However, the potential of HME5 as an antiproliferative and cytotoxic agent on an ovarian cancer cells has not yet been investigated. In this present study, we examined the antiproliferative and cytotoxic effect of HME5 on Caov-3 (Human Ovarian Adenocarcinoma) cell line by using 3-[4,5-dimethylthizol-2-y]-2,5-diphenyltetrazolium bromide (MTT) assay, Acridine orange and propidium Iodide (AOPi) and cell cycle analysis study. HME5 has shown to inhibit Caov-3 in a time-dependent manner with the IC50 values of 5µg/ml, 2µg/ml and 1µg/ml after 24h, 48h and 72h treatment, respectively. Morphological study from AOPi analysis showed that HME5 induced apoptosis after 24 and 48h post-treatment. Nevertheless, HME5 exhibited cell cycle arrest at G1 phase as indicated in flow cytometry cell cycle profiling. In conclusion, HME5 inhibited proliferation of Caov-3 through induction of apoptosis and cell cycle arrest at G1 phase.

Keywords: apoptosis, prenylflavanoid, ovarian cancer, HME5

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Authors: Z. Herceg, V. Stulic, T. Vukusic, A. Rezek Jambrak

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Gas phase plasma treatment is a new nonthermal technology used for food and water decontamination. In this study, we have investigated influence of the gas phase plasma treatment on yeast cells of S. cerevisiae. Sample was composed of 10 mL of yeast suspension and 190 mL of 0.01 M NaNO₃ with a medium conductivity of 100 µS/cm. Samples were treated in a glass reactor with a point- to-plate electrode configuration (high voltage electrode-titanium wire in the gas phase and grounded electrode in the liquid phase). Air or argon were injected into the headspace of the reactor at the gas flow of 5 L/min. Frequency of 60, 90 and 120 Hz, time of 5 and 10 min and positive polarity were defined parameters. Inactivation was higher with the applied higher frequency, longer treatment time and injected argon. Inactivation was not complete which resulted in complete recovery. Cellular leakage (260 nm and 280 nm) was higher with a longer treatment time and higher frequency. Leakage at 280 nm which defines a leakage of proteins was higher than leakage at 260 nm which defines a leakage of nucleic acids. The authors would like to acknowledge the support by Croatian Science Foundation and research project 'Application of electrical discharge plasma for preservation of liquid foods'.

Keywords: Saccharomyces cerevisiae, inactivation, gas-phase plasma treatment, cellular leakage

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Authors: Sven Borén, Henrik Ny

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Ambitions within the EU for moving towards sustainable transport include major emission reductions for fossil fuel road vehicles, especially for buses, trucks, and cars. The electric driveline seems to be an attractive solution for such development. This study first applied the Framework for Strategic Sustainable Development to compare sustainability effects of today’s fossil fuel vehicles with electric vehicles that have batteries or hydrogen fuel cells. The study then addressed a scenario were electric vehicles might be in majority in Europe by 2050. The methodology called Strategic Lifecycle Assessment was first used, were each life cycle phase was assessed for violations against sustainability principles. This indicates where further analysis could be done in order to quantify the magnitude of each violation, and later to create alternative strategies and actions that lead towards sustainability. A Life Cycle Assessment of combustion engine cars, plug-in hybrid cars, battery electric cars and hydrogen fuel cell cars was then conducted to compare and quantify environmental impacts. The authors found major violations of sustainability principles like use of fossil fuels, which contribute to the increase of emission related impacts such as climate change, acidification, eutrophication, ozone depletion, and particulate matters. Other violations were found, such as use of scarce materials for batteries and fuel cells, and also for most life cycle phases for all vehicles when using fossil fuel vehicles for mining, production and transport. Still, the studied current battery and hydrogen fuel cell cars have less severe violations than fossil fuel cars. The life cycle assessment revealed that fossil fuel cars have overall considerably higher environmental impacts compared to electric cars as long as the latter are powered by renewable electricity. By 2050, there will likely be even more sustainable alternatives than the studied electric vehicles when the EU electricity mix mainly should stem from renewable sources, batteries should be recycled, fuel cells should be a mature technology for use in vehicles (containing no scarce materials), and electric drivelines should have replaced combustion engines in other sectors. An uncertainty for fuel cells in 2050 is whether the production of hydrogen will have had time to switch to renewable resources. If so, that would contribute even more to a sustainable development. Except for being adopted in the GreenCharge roadmap, the authors suggest that the results can contribute to planning in the upcoming decades for a sustainable increase of EVs in Europe, and potentially serve as an inspiration for other smaller or larger regions. Further studies could map the environmental effects in LCA further, and include other road vehicles to get a more precise perception of how much they could affect sustainable development.

Keywords: strategic, electric vehicles, sustainability, LCA

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Authors: Eirini Krasaki

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This paper addresses the problem of designing spaces in a consistently changing environment. Space is considered as a totality of forces that coexist in the same place. Forces form the identity of space and characterize the entities that coexist within the same totality. Interior space is considered as a totality of forces which develop within an envelope. This research focuses on the formation of the tripole space-forces-totality and studies the relation of this tripole to the interior space. The point of departure for this investigation has been set the historic center of Athens, a city center where the majority of building mass is unused. The objective of the study is to connect the development of interior spaces to the alterations of the conceptions that form the built environment. The research focuses on Evripidou street, an axis around which expand both commercial and residential centers. Along Evripidou street, three case studies elaborate: a) In case study 01, Evripidou street is examined as a megastructure in which totalities of interior spaces develop. b) In case study 02, a particular group of entities (polykatoikia) that expand in Evripidou street is investigated. c) In case study 03, a particular group of entities (apartment) that derives from a specific envelope is investigated. Throughout the studies and comparisons of different scales, a design methodology that addresses the design of interior space in relation to the dynamics of the built environment is evolved.

Keywords: methodology, research by design, interior, envelope, dynamics

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Authors: Pouya Karimi, Ramin Gasemi Shayan, Parsa Sheykhzade

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Ease shotgun DNA sequencing is changing the microbial sciences. Sequencing instruments are compelling to the point that example planning is currently the key constraining element. Here, we present a microfluidic test readiness stage that incorporates the key strides in cells to grouping library test groundwork for up to 96 examples and decreases DNA input prerequisites 100-overlay while keeping up or improving information quality. The universally useful microarchitecture we show bolsters work processes with subjective quantities of response and tidy up or catch steps. By decreasing the example amount necessities, we empowered low-input (∼10,000 cells) entire genome shotgun (WGS) sequencing of Mycobacterium tuberculosis and soil miniaturized scale settlements with prevalent outcomes. We additionally utilized the upgraded throughput to succession ∼400 clinical Pseudomonas aeruginosa libraries and exhibit magnificent single-nucleotide polymorphism discovery execution that clarified phenotypically watched anti-toxin opposition. Completely coordinated lab-on-chip test arrangement beats specialized boundaries to empower more extensive organization of genomics across numerous fundamental research and translational applications.

Keywords: clinical microbiology, DNA, microbiology, microbial genomics

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Authors: Matej Komanec, Jan Bohata, Stanislav Zvanovec, Tomas Nemecek, Jan Broucek, Josef Beran

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With the exponential growth of social networks, video streaming and increasing demands on data rates, the number of newly built data centers rises proportionately. The data centers, however, have to adjust to the rapidly increased amount of data that has to be processed. For this purpose, multi-mode (MM) fiber based infrastructures are often employed. It stems from the fact, the connections in data centers are typically realized within a short distance, and the application of MM fibers and components considerably reduces costs. On the other hand, the usage of MM components brings specific requirements for installation service conditions. Moreover, it has to be taken into account that MM fiber components have a higher production tolerance for parameters like core and cladding diameters, eccentricity, etc. Due to the high demands for the reliability of data center components, the determination of properly excited optical field inside the MM fiber core belongs to the key parameters while designing such an MM optical system architecture. Appropriately excited mode field of the MM fiber provides optimal power budget in connections, leads to the decrease of insertion losses (IL) and achieves effective modal bandwidth (EMB). The main parameter, in this case, is the encircled flux (EF), which should be properly defined for variable optical sources and consequent different mode-field distribution. In this paper, we present detailed investigation and measurements of the mode field distribution for short MM links purposed in particular for data centers with the emphasis on reliability and safety. These measurements are essential for large MM network design. The various scenarios, containing different fibers and connectors, were tested in terms of IL and mode-field distribution to reveal potential challenges. Furthermore, we focused on estimation of particular defects and errors, which can realistically occur like eccentricity, connector shifting or dust, were simulated and measured, and their dependence to EF statistics and functionality of data center infrastructure was evaluated. The experimental tests were performed at two wavelengths, commonly used in MM networks, of 850 nm and 1310 nm to verify EF statistics. Finally, we provide recommendations for data center systems and networks, using OM3 and OM4 MM fiber connections.

Keywords: optical fiber, multi-mode, data centers, encircled flux

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Authors: Belal Bakheet, John Beardall, Xiwang Zhang, David McCarthy

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The potential risks associated with toxic cyanobacteria have raised growing environmental and public health concerns leading to an increasing effort into researching ways to bring about their removal from water, together with destruction of their associated cyanotoxins. A variety of toxins are synthesized by cyanobacteria and include hepatotoxins, neurotoxins, and cytotoxins which can cause a range of symptoms in humans from skin irritation to serious liver and nerve damage. Therefore drinking water treatment processes should ensure the consumers’ safety by removing both cyanobacterial cells, and cyanotoxins from the water. Cyanobacterial cells and cyanotoxins presented challenges to the conventional water treatment systems; their accumulation within drinking water treatment plants has been reported leading to plants shut down. Thus, innovative and effective water purification systems to tackle cyanobacterial pollution are required. In recent years there has been increasing attention to the electrochemical oxidation process as a feasible alternative disinfection method which is able to generate in situ a variety of oxidants that would achieve synergistic effects in the water disinfection process and toxin degradation. By utilizing only electric current, the electrochemical process through electrolysis can produce reactive oxygen species such as hydroxyl radicals from the water, or other oxidants such as chlorine from chloride ions present in the water. From extensive physiological and morphological investigation of cyanobacterial cells during electrolysis, our results show that these oxidants have significant impact on cell inactivation, simultaneously with cyanotoxins removal without the need for chemicals addition. Our research aimed to optimize existing electrochemical oxidation systems and develop new systems to treat water containing toxic cyanobacteria and cyanotoxins. The research covers detailed mechanism study on oxidants production and cell inactivation in the treatment under environmental conditions. Overall, our study suggests that the electrochemical treatment process e is an effective method for removal of toxic cyanobacteria and cyanotoxins.

Keywords: toxic cyanobacteria, cyanotoxins, electrochemical process, oxidants

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Authors: Maria L. McLeod

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This study examines the ways in which cancer patient narratives are portrayed and framed on the websites of three leading U.S. cancer care centers –The University of Texas MD Anderson Cancer Center in Houston, Memorial Sloan Kettering Cancer Center in New York, and Seattle Cancer Care Alliance. Thirty patient stories, ten from each cancer center website blog, were analyzed using qualitative and quantitative textual analysis of unstructured data, documenting repeated use of specific metaphors and tropes while charting common themes and other elements of story structure and content. Patient narratives were coded using grounded theory as the basis for conducting emergent qualitative research. As part of a systematic, inductive approach to collecting and analyzing data, recurrent and unique themes were examined and compared in terms of positive and negative framing, patient agency, and institutional praise. All three of these cancer care centers are teaching hospitals with university affiliations, that emphasizes an evidence-based scientific approach to treatment that utilizes the latest research and cutting-edge techniques and technology. Thus, the use of anecdotal evidence presented in patient narratives could be perceived as being in conflict with this evidence-based model, as the patient stories are not an accurate representation of scientific outcomes related to developing cancer, cancer reoccurrence, or cancer outcomes. The representative patient narratives tend to exclude or downplay adverse responses to treatment, survival rates, integrative and/or complementary cancer treatments, cancer prevention and causes, and barriers to treatment, such as the limitation of insurance plans, costs of treatment, and/or other issues related to access, potentially contributing to false narratives and inaccurate notions of cancer prevention, cancer care treatment and the potential for a cure. Both quantitative and qualitative findings demonstrate that cancer patient stories featured on the blogsites of the nation’s top cancer care centers deemphasize patient agency and, instead, emphasize deference and gratitude toward the institutions where the featured patients received treatment. Along these lines, language choices reflect positive framing of the cancer experience. Accompanying portrait photos of healthy appearing subjects as well as positive-framed headlines, subheads, and pull quotes function similarly, reflecting hopeful, transformative experiences and outcomes over hardship and suffering. Although patient narratives include real, factual scientific details and descriptions of actual events, the stories lack references to more negative realities of cancer diagnosis and treatment. Instead, they emphasize the triumph of survival by which the cancer care center, in the savior/hero role, enables the patient’s success, represented as a cathartic medical journey.

Keywords: cancer framing, cancer stories, medical gaze, patient narratives

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Authors: Congping Lin

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Intracellular transport in fungi has a number of important roles in, e.g., filamentous fungal growth and cellular metabolism. Two basic mechanisms for intracellular transport are motor-driven trafficking along microtubules (MTs) and diffusion. Mathematical modelling has been actively developed to understand such intracellular transport and provide unique insight into cellular complexity. Based on live-cell imaging data in Ustilago hyphal cells, probabilistic models have been developed to study mechanism underlying spatial organization of molecular motors and organelles. In particular, anther mechanism - stochastic motility of dynein motors along MTs has been found to contribute to half of its accumulation at hyphal tip in order to support early endosome (EE) recycling. The EE trafficking not only facilitates the directed motion of peroxisomes but also enhances their diffusive motion. Considering the importance of spatial organization of early endosomes in supporting peroxisome movement, computational and experimental approaches have been combined to a whole-cell level. Results from this interdisciplinary study promise insights into requirements for other membrane trafficking systems (e.g., in neurons), but also may inform future 'synthetic biology' studies.

Keywords: intracellular transport, stochastic process, molecular motors, spatial organization

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Authors: Surekha Barkur, Aseefhali Bankapur, Santhosh Chidangil

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Raman Tweezers technique has become prevalent in single cell studies. This technique combines Raman spectroscopy which gives information about molecular vibrations, with optical tweezers which use a tightly focused laser beam for trapping the single cells. Thus Raman Tweezers enabled researchers analyze single cells and explore different applications. The applications of Raman Tweezers include studying blood cells, monitoring blood-related disorders, silver nanoparticle-induced stress, etc. There is increased interest in the toxic effect of nanoparticles with an increase in the various applications of nanoparticles. The interaction of these nanoparticles with the cells may vary with their size. We have studied the effect of silver nanoparticles of sizes 10nm, 40nm, and 100nm on erythrocytes using Raman Tweezers technique. Our aim was to investigate the size dependence of the nanoparticle effect on RBCs. We used 785nm laser (Starbright Diode Laser, Torsana Laser Tech, Denmark) for both trapping and Raman spectroscopic studies. 100 x oil immersion objectives with high numerical aperture (NA 1.3) is used to focus the laser beam into a sample cell. The back-scattered light is collected using the same microscope objective and focused into the spectrometer (Horiba Jobin Vyon iHR320 with 1200grooves/mm grating blazed at 750nm). Liquid nitrogen cooled CCD (Symphony CCD-1024x256-OPEN-1LS) was used for signal detection. Blood was drawn from healthy volunteers in vacutainer tubes and centrifuged to separate the blood components. 1.5 ml of silver nanoparticles was washed twice with distilled water leaving 0.1 ml silver nanoparticles in the bottom of the vial. The concentration of silver nanoparticles is 0.02mg/ml so the 0.03mg of nanoparticles will be present in the 0.1 ml nanoparticles obtained. The 25 ul of RBCs were diluted in 2 ml of PBS solution and then treated with 50 ul (0.015mg) of nanoparticles and incubated in CO2 incubator. Raman spectroscopic measurements were done after 24 hours and 48 hours of incubation. All the spectra were recorded with 10mW laser power (785nm diode laser), 60s of accumulation time and 2 accumulations. Major changes were observed in the peaks 565 cm-1, 1211 cm-1, 1224 cm-1, 1371 cm-1, 1638 cm-1. A decrease in intensity of 565 cm-1, increase in 1211 cm-1 with a reduction in 1224 cm-1, increase in intensity of 1371 cm-1 also peak disappearing at 1635 cm-1 indicates deoxygenation of hemoglobin. Nanoparticles with higher size were showing maximum spectral changes. Lesser changes observed in case of 10nm nanoparticle-treated erythrocyte spectra.

Keywords: erythrocytes, nanoparticle-induced toxicity, Raman tweezers, silver nanoparticles

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Authors: Felsy J. Sandi

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There is a potential of growth in the spa industry due to the influx of domestic and international tourist coming to Sabah, Malaysia. It is a good opportunity to venture into this industry for the country’s economic future growth, and therefore, it is essential for this area to be researched. Being one of the fastest growing industries in the world, has led to enormous challenges, which need to be addressed. Malaysia is also riding with this phenomenon. The President of the Malaysian Association of Wellness and Spa stated that the misconception about the Spa industry’s image, especially amongst the elderly is the biggest challenge faced by the industry, as they perceived the spa industry is equivalent to a prostitution center. Therefore, the objective of this study is to explore the issue by analyzing whether image can be added in the theory of planned behavior to better understand the consumer’s intention to visit, in the spa context. The Theory of Planned Behavior by Ajzen, a theory or model in predicting intention, has three constructs; such as Attitude as the first construct, the second construct is Subjective Norm and the third construct is Perceived Behavioral Control. Qualitative research is used as this is an exploratory research. The site of study will be at Jari Jari Spa, located in Kota Kinabalu, the only spa in Sabah that was awarded as the Center of Excellence (CoE) by the Ministry of Tourism and Culture in Malaysia. The findings propose to provide useful information to the relevant stakeholders on ways to approach local customers to convince them to visit the spa and for spa marketers to help them develop and design effective marketing strategies. Future investigation should consider more on the perception and loyalty of the local customers.

Keywords: consumer's perception, image, local customer, spa, visit intention

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Authors: Zeinab Hajifathali, Moghan Amirhosseinian

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This study had two main aims: firstly, to determine how the individual substitution of CaO/MgO and CaO/SrO can affect the in vitro bioactivity of sol-gel derived substituted 58S bioactive glass (BG) and secondly to introduce a composition in the 60SiO₂–(36-x)CaO–4P₂O₅–(x)MgO and 60SiO₂–(36-x)CaO–4P₂O₅–(x)SrO quaternary systems (where x= 0, 5, 10 mol.%) with enhanced biocompatibility, alkaline phosphatase (ALP) activity, and more efficient antibacterial activity against MRSA bacteria. Results showed that both magnesium-substituted bioactive glasses (M-BGs) and strontium- substituted bioactive glasses (S-BGs) retarded the Hydroxyapatite (HA) formation. Meanwhile, magnesium had more pronounced effect. The 3-(4, 5dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and ALP assays revealed that the presence of moderate amount (5 mol%) of Mg and Sr had a stimulating effect on increasing of both proliferation and differentiation of MC3T3-E1 cells. Live dead and Dapi/actin staining revealed both substitution of CaO/MgO and CaO/SrO resulted in more biocompatibility and stimulation potential of the MC3T3 cells compared with control. Taken together, among all of the synthesized magnesium substituted (MBGs) and strontium substituted (SBGs), the sample 58- BG with 5 mol% CaO/MgO substitution (BG-5M) was considered as a multifunctional biomaterial in bone tissue regeneration field with enhanced biocompatibility, ALP activity as well as the highest antibacterial efficiency against methicillin-resistant Staphylococcus aureus (MRSA) bacteria.

Keywords: apatite, alkaline earth, bioactivity, biomedical applications, Sol-gel

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Authors: Fadi Saleh, Çığdem Sezer Zhmurov

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Biopharmaceuticals represent one of the wildest developing fields within biotechnology, and the biological macromolecules being produced inside cells have a variety of applications for therapies. In the past, mammalian cells, especially CHO cells, have been employed in the production of biopharmaceuticals. This is because these cells can achieve human-like completion of PTM. These systems, however, carry apparent disadvantages like high production costs, vulnerability to contamination, and limitations in scalability. This research is focused on the utilization of microalgae as a bioreactor system for the synthesis of biopharmaceutical glycoproteins in relation to PTMs, particularly N-glycosylation. The research points to a growing interest in microalgae as a potential substitute for more conventional expression systems. A number of advantages exist in the use of microalgae, including rapid growth rates, the lack of common human pathogens, controlled scalability in bioreactors, and the ability of some PTMs to take place. Thus, the potential of microalgae to produce recombinant proteins with favorable characteristics makes this a promising platform in order to produce biopharmaceuticals. The study focuses on the examination of the N-glycosylation pathways across different species of microalgae. This investigation is important as N-glycosylation—the process by which carbohydrate groups are linked to proteins—profoundly influences the stability, activity, and general performance of glycoproteins. Additionally, bioinformatics methodologies are employed to explain the genetic pathways implicated in N-glycosylation within microalgae, with the intention of modifying these organisms to produce glycoproteins suitable for human consumption. In this way, the present comparative analysis of the N-glycosylation pathway in humans and microalgae can be used to bridge both systems in order to produce biopharmaceuticals with humanized glycosylation profiles within the microalgal organisms. The results of the research underline microalgae's potential to help improve some of the limitations associated with traditional biopharmaceutical production systems. The study may help in the creation of a cost-effective and scale-up means of producing quality biopharmaceuticals by modifying microalgae genetically to produce glycoproteins with N-glycosylation that is compatible with humans. Improvements in effectiveness will benefit biopharmaceutical production and the biopharmaceutical sector with this novel, green, and efficient expression platform. This thesis, therefore, is thorough research into the viability of microalgae as an efficient platform for producing biopharmaceutical glycoproteins. Based on the in-depth bioinformatic analysis of microalgal N-glycosylation pathways, a platform for their engineering to produce human-compatible glycoproteins is set out in this work. The findings obtained in this research will have significant implications for the biopharmaceutical industry by opening up a new way of developing safer, more efficient, and economically more feasible biopharmaceutical manufacturing platforms.

Keywords: microalgae, glycoproteins, post-translational modification, genome

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

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

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

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Authors: Tomilola J. Ajayi, Moses Ollengo, Lukas le Roux, Michael N. Pillay, Richard J. Staples, Shannon M. Biros Werner E. van Zyl

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The formation, characterization, and dye-sensitized solar cell application of nickel(II), zinc(II) and cadmium(II) ferrocenyl dithiophosphonate complexes were investigated. The multidentate monoanionic ligand [S₂PFc(OH)]¯ (L1) was synthesized from the reaction between ferrocenyl Lawesson’s reagent, [FcP(=S)μ-S]₂ (FcLR), (Fc = ferrocenyl) and water. Ligand L1 could potentially coordinate to metal centers through the S, S’ and O donor atoms. The reaction between metal salt precursors and L1 produced a Ni(II) complex of the type [Ni{S₂P(Fc)(OH)}₂] (1) (molar ratio 1:2), a tetranickel (II) complex of the type [Ni₂{S₂OP(Fc)}₂]₂ (2) (molar ratio (1:1), as well as a Zn(II) complex [Zn{S₂P(Fc)(OH)}₂]₂ (3), and a Cd(II) complex [Cd{S₂P(Fc)(OH)}₂]₂ (4). Complexes 1-4 were characterized by 1H and 31P NMR and FT-IR, and complexes 1 and 2 were additionally analysed by X-Ray crystallography. After co-sensitization, the DSSCs were characterized using UV-Vis, cyclic voltammetry, electrochemical impedance spectroscopy, and photovoltaic measurements (I-V curves). Overall finding shows that co-sensitization of our compounds with ruthenium dye N719 resulted in a better overall solar conversion efficiency than only pure N719 dye under the same experimental conditions. In conclusion, we report the first examples of dye-sensitized solar cells (DSSCs) co-sensitized with ferrocenyl dithiophosphonate complexes.

Keywords: dithiophosphonate, dye sensitized solar cell, co-sensitization, solar efficiency

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Authors: Subal Kar, Amitesh Kumar, A. Majumder, S. K. Ghosh, S. Saha, S. S. Sikdar, T. K. Saha

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CRLH (composite right/left-handed) based and SRR (split-ring resonator) based filters have been designed at microwave frequency which can provide better performance compared to conventional edge-coupled band-pass filter designed around the same frequency, 2.45 GHz. Both CRLH and SRR are unit cells used in metamaterial design. The primary aim of designing filters with such structures is to realize size reduction and also to realize novel filter performance. The CRLH based filter has been designed in microstrip transmission line, while the SRR based filter is designed with SRR loading in waveguide. The CRLH based filter designed at 2.45 GHz provides an insertion loss of 1.6 dB with harmonic suppression up to 10 GHz with 67 % size reduction when compared with a conventional edge-coupled band-pass filter designed around the same frequency. One dimensional (1-D) SRR matrix loaded in a waveguide shows the possibility of realizing a stop-band with sharp skirts in the pass-band while a stop-band in the pass-band of normal rectangular waveguide with tailoring of the dimensions of SRR unit cells. Such filters are expected to be very useful for communication systems at microwave frequency.

Keywords: BPF, CRLH, harmonic, metamaterial, SRR and waveguide

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Authors: Kunyaporn Pipithsangchan, Alongkorn Korntong, Assanee Songserm, Phatchara Piriyavinit, Saowanee Dechakampoo

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The Kingdom of Thailand is one of the South East Asian countries. From its area of 514,000 square kilometers (51 million ha), at least 18,000 plant species (8% of the world total) have been estimated to be found in the country. As a result, the conservation of plant genetic diversity, particularly food crops, is becoming important and is an assurance for the national food security. Department of Agriculture Genebank or DOA Genebank, Thailand is responsible for the conservation of plant germplasm by participating and accomplishing several collaborative projects both at national and international levels. Integrated Management System of Plant Genetic Resources or IMPGR is one of the most outstandingly successful cooperation. It is a multilateral project under the Asian Food and Agriculture Cooperation Initiative (AFACI) supported by the Rural Development Administration (RDA) of South Korea. The member countries under the project consist of 11 nations namely Bangladesh, Cambodia, Indonesia, Laos PDR, Mongolia, Nepal, Philippines, Sri Lanka, Thailand, Vietnam and South Korea. The project enabled the members to jointly address the global issues in plant genetic resource (PGR) conservation and strengthen their network in this aspect. The 1st phase of IMPGR project, entitled 'Collection, Conservation, Regeneration and Characterization of Cucurbitaceae and Solanaceae 2012-2014', comprises three main objectives that are: 1) To improve management in storage facilities, collection, and regeneration, 2) To improve linkage between Genebank and material sources (for regeneration), and 3) To improve linkage between Genebank and other field crop or/and horticultural research centers. The project was done for three years from 2012 to 2014. The activities of the project can be described as following details: In the 1st year, there were 9 target provinces for completing plant genetic resource survey and collection. 108 accessions of PGR were collected. In the 2nd year, PGR were continuously surveyed and collected from 9 provinces. The total number of collection was 140 accessions. In addition, the process of regeneration of 237 accessions collected from 1st and 2nd year was started at several sites namely Biotechnology Research and Development Office, Sukothai Horticultural Research Center, Tak Research, and Development Center and Nakhon Ratchasima Research and Development Center. In the 3rd year, besides survey and collection of 115 accessions from 9 target provinces, PGR characterization and evaluation were done for 206 accessions. Moreover, safety duplication of 253 PGR at the World Seed Vault, RDA, was also done according to Standard Agreement on Germplasm Safety Duplication between Department of Agriculture, Ministry of Agriculture and Cooperatives, the Kingdom of Thailand and the National Agrobiodiversity Center, Rural Development Administration of the Republic of Korea. The success of the 1st phase project led to the second phase which entitled 'Collection and Characterization for Effective Conservation of Local Capsicum spp., Solanum spp. and Lycopersicon spp. in Thailand 2015-2017'.

Keywords: characterization, conservation, DOA genebank, plant genetic resources

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Authors: Samia A. El-Fiky, Fouad A. Abou-Zaid, Ibrahim M. Farag, Naira M. El-Fiky

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Clomipramine hydrochloride is one of the most used tricyclic antidepressant drug in Egypt. This drug contains in its chemical structure on two benzene rings. Benzene is considered to be toxic and clastogenic agent. So, the present study was designed to assess the genotoxic effect of Clomipramine hydrochloride on somatic and germ cells in mice. Three dose levels 0.195 (Low), 0.26 (Medium), and 0.65 (High) mg/kg.b.wt. were used. Seven groups of male mice were utilized in this work. The first group was employed as a control. In the remaining six groups, each of the above doses was orally administrated for two groups, one of them was treated for 5 days and the other group was given the same dose for 30 days. At the end of experiments, the animals were sacrificed for cytogenetic and sperm examination as well as histopathological investigations by using hematoxylin and eosin stains (H and E stains) and electron microscope. Concerning the sperm studies, these studies were confined to 5 days treatment with different dose levels. Moreover, the ultrastructural investigation by electron microscope was restricted to 30 days treatment with drug doses. The results of the dose dependent effect of Clomipramine showed that the treatment with three different doses induced increases of frequencies of chromosome aberrations in bone marrow and spermatocyte cells as compared to control. In addition, mitotic and meiotic activities of somatic and germ cells were declined. The treatments with medium or high doses were more effective for inducing significant increases of chromosome aberrations and significant decreases of cell divisions than treatment with low dose. The effect of high dose was more pronounced for causing such genetic deleterious in respect to effect of medium dose. Moreover, the results of the time dependent effect of Clomipramine observed that the treatment with different dose levels for 30 days led to significant increases of genetic aberrations than treatment for 5 days. Sperm examinations revealed that the treatment with Clomipramine at different dose levels caused significant increase of sperm shape abnormalities and significant decrease in sperm count as compared to control. The adverse effects on sperm shape and count were more obviousness by using the treatments with medium or high doses than those found in treatment with low dose. The group of mice treated with high dose had the highest rate of sperm shape abnormalities and the lowest proportion of sperm count as compared to mice received medium dose. In histopathological investigation, hematoxylin and eosin stains showed that, the using of low dose of Clomipramine for 5 or 30 days caused a little pathological changes in liver tissue. However, using medium and high doses for 5 or 30 days induced severe damages than that observed in mice treated with low dose. The treatment with high dose for 30 days gave the worst results of pathological changes in hepatic cells. Moreover, ultrastructure examination revealed, the mice treated with low dose of Clomipramine had little differences in liver histological architecture as compared to control group. These differences were confined to cytoplasmic inclusions. Whereas, prominent pathological changes in nuclei as well as dilated of rough Endoplasmic Reticulum (rER) were observed in mice treated with medium or high doses of Clomipramine drug. In conclusion, the present study adds evidence that treatments with medium or high doses of Clomipramine have genotoxic effects on somatic and germ cells of mice, as unwanted side effects. However, the using of low dose (especially for short time, 5 days) can be utilized as a therapeutic dose, where it caused relatively similar proportions of genetic, sperm, and histopathological changes as those found in normal control.

Keywords: chromosome aberrations, clomipramine, mice, histopathology, sperm abnormalities

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Authors: Michal Kielbik, Izabela Szulc-Kielbik, Anna Brzostek, Jaroslaw Dziadek, Magdalena Klink

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The goal of many research groups in the world is to find new components that are important for survival of mycobacteria in the host cells. Mycobacterium tuberculosis (Mtb) possesses a number of enzymes degrading cholesterol that are considered to be an important factor for its survival and persistence in host macrophages. One of them - cholesterol oxidase (ChoD), although not being essential for cholesterol degradation, is discussed as a virulence compound, however its involvement in macrophages’ response to Mtb is still not sufficiently determined. The recognition of tubercle bacilli antigens by pathogen recognition receptors is crucial for the initiation of the host innate immune response. An important receptor that has been implicated in the recognition and/or uptake of Mtb is Toll-like receptor type 2 (TLR2). Engagement of TLR2 results in the activation and phosphorylation of intracellular signaling proteins including IRAK-1 and -4, TRAF-6, which in turn leads to the activation of target kinases and transcription factors responsible for bactericidal and pro-inflammatory response of macrophages. The aim of these studies was a detailed clarification of the role of Mtb cholesterol oxidase as a virulence factor affecting the TLR2 signaling pathway in human macrophages. As human macrophages the THP-1 differentiated cells were applied. The virulent wild-type Mtb strain (H37Rv), its mutant lacking a functional copy of gene encoding cholesterol oxidase (∆choD), as well as complimented strain (∆choD–choD) were used. We tested the impact of Mtb strains on the expression of TLR2-depended signaling proteins (mRNA level, cytosolic level and phosphorylation status). The cytokine and bactericidal response of THP-1 derived macrophages infected with Mtb strains in relation to TLR2 signaling pathway dependence was also determined. We found that during the 24-hours of infection process the wild-type and complemented Mtb significantly reduced the cytosolic level and phosphorylation status of IRAK-4 and TRAF-6 proteins in macrophages, that was not observed in the case of ΔchoD mutant. Decreasement of TLR2-dependent signaling proteins, induced by wild-type Mtb, was not dependent on the activity of proteasome. Blocking of TLR2 expression, before infection, effectively prevented the induced by wild-type strain reduction of cytosolic level and phosphorylation of IRAK-4. None of the strains affected the surface expression of TLR2. The mRNA level of IRAK-4 and TRAF-6 genes were significantly increased in macrophages 24 hours post-infection with either of tested strains. However, the impact of wild-type Mtb strain on both examined genes was significantly stronger than its ΔchoD mutant. We also found that wild-type strain stimulated macrophages to release high amount of immunosuppressive IL-10, accompanied by low amount of pro-inflammatory IL-8 and bactericidal nitric oxide in comparison to mutant lacking cholesterol oxidase. The influence of wild-type Mtb on this type of macrophages' response strongly dependent on fully active IRAK-1 and IRAK-4 signaling proteins. In conclusion, Mtb using cholesterol oxidase causes the over-activation of TLR2 signaling proteins leading to the reduction of their cytosolic level and activity resulting in the modulation of macrophages response to allow its intracellular survival. Supported by grant: 2014/15/B/NZ6/01565, National Science Center, Poland

Keywords: Mycobacterium tuberculosis, cholesterol oxidase, macrophages, TLR2-dependent signaling pathway

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Authors: Sadaf Ilyas

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

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

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Authors: Leo Nnamdi Ozurumba-Dwight

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Messenger ribooxynucleic acid (mRNA) molecules are compositional, protein-based. These proteins, encoding mRNA molecules (which collectively connote the transcriptome), when analyzed by RNA sequencing (RNAseq), unveils the nature of gene expression in the RNA. The obtained gene expression provides clues of cellular traits and their dynamics in presentations. These can be studied in relation to function and responses. RNAseq is a practical concept in Genomics as it enables detection and quantitative analysis of mRNA molecules. Single cell and spatial transcriptomics both present varying avenues for expositions in genomic characteristics of single cells and pooled cells in disease conditions such as cancer, auto-immune diseases, hematopoietic based diseases, among others, from investigated biological tissue samples. Single cell transcriptomics helps conduct a direct assessment of each building unit of tissues (the cell) during diagnosis and molecular gene expressional studies. A typical technique to achieve this is through the use of a single-cell RNA sequencer (scRNAseq), which helps in conducting high throughput genomic expressional studies. However, this technique generates expressional gene data for several cells which lack presentations on the cells’ positional coordinates within the tissue. As science is developmental, the use of complimentary pre-established tissue reference maps using molecular and bioinformatics techniques has innovatively sprung-forth and is now used to resolve this set back to produce both levels of data in one shot of scRNAseq analysis. This is an emerging conceptual approach in methodology for integrative and progressively dependable transcriptomics analysis. This can support in-situ fashioned analysis for better understanding of tissue functional organization, unveil new biomarkers for early-stage detection of diseases, biomarkers for therapeutic targets in drug development, and exposit nature of cell-to-cell interactions. Also, these are vital genomic signatures and characterizations of clinical applications. Over the past decades, RNAseq has generated a wide array of information that is igniting bespoke breakthroughs and innovations in Biomedicine. On the other side, spatial transcriptomics is tissue level based and utilized to study biological specimens having heterogeneous features. It exposits the gross identity of investigated mammalian tissues, which can then be used to study cell differentiation, track cell line trajectory patterns and behavior, and regulatory homeostasis in disease states. Also, it requires referenced positional analysis to make up of genomic signatures that will be sassed from the single cells in the tissue sample. Given these two presented approaches to RNA transcriptomics study in varying quantities of cell lines, with avenues for appropriate resolutions, both approaches have made the study of gene expression from mRNA molecules interesting, progressive, developmental, and helping to tackle health challenges head-on.

Keywords: transcriptomics, RNA sequencing, single cell, spatial, gene expression.

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Authors: Marzieh Amiri, Zahra Iranpour Mobarakeh, Fatemeh Mehrbakhsh, Mehran Amiri

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Introduction: Early diagnosis and intervention of congenital hearing loss is necessary to minimize the adverse effects of hearing loss. The aim of the present study was to report the results of newborn hearing screening in a centerin the south part of Iran, Fasa. Material and methods: In this study, the data related to 6,144 newbornsduring September 2018 up to September2021, was analyzed. Hearing screening was performed using transient evoked otoacoustic emissions (TEOAEs) and automated auditory brainstem response (AABR) tests. Results: From all 6144 newborns,3752 and 2392referred to the center from urban and rural part of Fasa, respectively. There were 2958 female and 3186 male in this study. Of 6144 newborns, 6098 ones passed the screening tests, and 46 neonates were referred to a diagnostic audiology clinic. Finally, nine neonates were diagnosed with congenital hearing loss (seven with sensorineural hearing loss and two with conductive hearing loss). The severity of all the hearing impaired neonates was moderate and above. The most important risk factors were family history of hearing loss, low gestational age, NICU hospitalization, and hyperbilirubinemia. Conclusion: Our results showed that the prevalence of hearing loss was 1.46 per 1000 infants. Boosting public knowledge by providing families with proper education appears to be helpful in preventing the negative effects of delayed implementation of health screening programs.

Keywords: newborn hearing screening, hearing loss, risk factor, prevalence

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Authors: Mohammad Alipour, Ekin Esen, Riza Kizilel

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Lithium-ion batteries are a commonly used type of rechargeable batteries because of their high specific energy and specific power. With the growing popularity of electric vehicles and hybrid electric vehicles, increasing attentions have been paid to rechargeable Lithium-ion batteries. However, safety problems, high cost and poor performance in low ambient temperatures and high current rates, are big obstacles for commercial utilization of these batteries. By proper thermal management, most of the mentioned limitations could be eliminated. Temperature profile of the Li-ion cells has a significant role in the performance, safety, and cycle life of the battery. That is why little temperature gradient can lead to great loss in the performances of the battery packs. In recent years, numerous researchers are working on new techniques to imply a better thermal management on Li-ion batteries. Keeping the battery cells within an optimum range is the main objective of battery thermal management. Commercial Li-ion cells are composed of several electrochemical layers each consisting negative-current collector, negative electrode, separator, positive electrode, and positive current collector. However, many researchers have adopted a single-layer cell to save in computing time. Their hypothesis is that thermal conductivity of the layer elements is so high and heat transfer rate is so fast. Therefore, instead of several thin layers, they model the cell as one thick layer unit. In previous work, we showed that single-layer model is insufficient to simulate the thermal behavior and temperature nonuniformity of the high-capacity Li-ion cells. We also studied the effects of the number of layers on thermal behavior of the Li-ion batteries. In this work, first thermal and electrochemical behavior of the LiFePO₄ battery is modeled with 3D multilayer cell. The model is validated with the experimental measurements at different current rates and ambient temperatures. Real time heat generation rate is also studied at different discharge rates. Results showed non-uniform temperature distribution along the cell which requires thermal management system. Therefore, aluminum plates with mini-channel system were designed to control the temperature uniformity. Design parameters such as channel number and widths, inlet flow rate, and cooling fluids are optimized. As cooling fluids, water and air are compared. Pressure drop and velocity profiles inside the channels are illustrated. Both surface and internal temperature profiles of single cell and battery packs are investigated with and without cooling systems. Our results show that using optimized Mini-channel cooling plates effectively controls the temperature rise and uniformity of the single cells and battery packs. With increasing the inlet flow rate, cooling efficiency could be reached up to 60%.

Keywords: lithium ion battery, 3D multilayer model, mini-channel cooling plates, thermal management

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Authors: Wooseok Byon, Eunyoung Kim, Junseong Kwon, Byung Joo Song, Chan Heun Park

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Background/Purpose: Previous preclinical and clinical data suggest that increased lymphocytic infiltration would be associated with good prognosis and benefit from immunogenic chemotherapy especially in triple-negative breast cancer (TNBC). We investigated a single-center experience of TNBC and relationship with lymphocytic infiltration. Methods: From January 2004 to December 2012, at the Department of Surgery, Kangbuk Samsung Hospital, Sungkyunkwan University, School of Medicine, we retrospectively reviewed 897 breast cancer patients-clinical outcomes, clinicopathological characteristics, breast cancer subtypes. And we reviewed lymphocytic infiltration of TNBC specimens by two pathologists. Statistical analysis of risk factors associated with recurrence was performed. Results: A total of 897 patients, 76 were TNBC (8.47%). Mean age of TNBC patients were 50.95 (SD10.42) years, mean follow-up periods was 40.06 months. We reviewed 49 slides, and there were 8 recurrent breast cancer patients (16.32%), and 4 patients were expired (8.16%). There were 9 lymphocytic predominant breast cancers (LPBC)-carcinomas with either intratumoral lymphocytes in >60% of tumor cell nests. 1 patient of LPBC was recurred and 8 were not. In multivariate logistic regression, the odds ratio of lymphocytic infiltration was 0.59 (p=0.643). Conclusion: In a single-center experience of TNBC, the lymphocytic infiltration in tumor cell nest might be a good trend on the prognosis but there was not statistically significant.

Keywords: tumor-infiltrating lymphocytes, triple negative breast cancer, medical and health sciences

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Authors: Chaiwat Boonkaewwan, Anutian Suklek, Jatuporn Rattanasrisomporn, Autchara Kayan

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Toll-like receptors (TLR) are conserved microbial sensing receptors located on cell surface that are able to detect different pathogens. The aim of the present study is to examine the expression of TLR gene in peripheral blood mononuclear cell of Betong (KU line) chicken. Blood samples were collected from healthy 12 Betong (KU line) chicken. PBMCs were isolated and maintained in RPMI1640 with 10% FBS, penicillin and streptomycin. Cell viability was determined by trypan blue dye exclusion test. The expression of TLRs gene was investigated by polymerase chain reaction (PCR) technique. Results showed that PBMCs viability from Betong (KU line) chicken was 95.38 ± 1.06%. From the study of TLRs gene expression, results indicated that there are expressions of TLR1.1 TLR1.2 TLR2.1 TLR2.2 TLR3 TLR4 TLR5 TLR 7 TLR15 and TLR21 in PBMCs of Betong (KU line) chicken. In conclusion, PBMCs isolated from blood of Betong (KU line) chicken had a high cell viability ( > 95%). The expression of TLRs in chicken was all found in PBMCs, which indicated that PBMC isolated from the blood of Betong (KU line) chicken can be used as an in vitro immune responses study.

Keywords: toll-like receptor, Betong (KU line) chicken, peripheral blood mononuclear cells

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Authors: Amir Peyman Soleymani, Jasna Jankovic

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The augmented demand of the clean and renewable energy has necessitated the fuel cell and battery industries to produce more efficient devices at the lower prices, which can be achieved through the improvement of the electrode. Microstructural characterization, as one of the main materials development tools, plays a pivotal role in the production of better clean energy devices. In this study, methods for characterization and studying of the defects and components distribution were performed on the polymer electrolyte membrane fuel cell (PEMFC) and Li-ion battery (LIB) electrodes in 2D and 3D. The particles distribution, porosity, mechanical defects, and component distribution were studied by Scanning Electron Microscope (SEM), SEM-Focused Ion Beam (SEM-FIB), and Scanning Transmission Electron Microscope equipped with Energy Dispersive Spectroscopy (STEM-EDS). The 3D results obtained from X-ray Computed Tomography (XCT) revealed the pathways for electron and ion conductivity and defects progression maps. Computer-aided methods (Avizo) were employed to simulate the properties and performance of the microstructure in the electrodes. The suggestions were provided to improve the performance of PEMFCs and LIBs by adjusting the microstructure and the distribution of the components in the electrodes.

Keywords: PEM fuel cells, Li-ion batteries, 2D and 3D imaging, materials characterizations

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Authors: Roxane A. Legaie, Kjiana E. Schwab, Caroline E. Gargett

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Studies looking at the changes in gene expression from RNAseq data often make use of linear models. It is also common practice to focus on a subset of data for a comparison of interest, leaving aside the samples not involved in this particular comparison. This work shows the importance of including all observations in the modeling process to better estimate variance parameters, even when the samples included are not directly used in the comparison under test. The human endometrium is a dynamic tissue, which undergoes cycles of growth and regression with each menstrual cycle. The mesenchymal stem cells (MSCs) present in the endometrium are likely responsible for this remarkable regenerative capacity. However recent studies suggest that MSCs also plays a role in the pathogenesis of endometriosis, one of the most common medical conditions affecting the lower abdomen in women in which the endometrial tissue grows outside the womb. In this study we compared gene expression profiles between MSCs and non-stem cell counterparts (‘non-MSC’) obtained from women with (‘E’) or without (‘noE’) endometriosis from RNAseq. Raw read counts were used for differential expression analysis using a linear model with the limma-voom R package, including either all samples in the study or only the samples belonging to the subset of interest (e.g. for the comparison ‘E vs noE in MSC cells’, including only MSC samples from E and noE patients but not the non-MSC ones). Using the full dataset we identified about 100 differentially expressed (DE) genes between E and noE samples in MSC samples (adj.p-val < 0.05 and |logFC|>1) while only 9 DE genes were identified when using only the subset of data (MSC samples only). Important genes known to be involved in endometriosis such as KLF9 and RND3 were missed in the latter case. When looking at the MSC vs non-MSC cells comparison, the linear model including all samples identified 260 genes for noE samples (including the stem cell marker SUSD2) while the subset analysis did not identify any DE genes. When looking at E samples, 12 genes were identified with the first approach and only 1 with the subset approach. Although the stem cell marker RGS5 was found in both cases, the subset test missed important genes involved in stem cell differentiation such as NOTCH3 and other potentially related genes to be used for further investigation and pathway analysis.

Keywords: differential expression, endometriosis, linear model, RNAseq

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Authors: Priyanka Bhowmik, Subrata Majumdar, Debprasad Chattopadhyay

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Background: Cervical cancer is the third major cause of cancer in women and the second most frequent cause of cancer related deaths causing 300,000 deaths annually worldwide. Evasion of immune response by Human Papilloma Virus (HPV), the key contributing factor behind cancer and pre-cancerous lesions of the uterine cervix, makes immunotherapy a necessity to treat this disease. Objective: A Heat killed fraction of Mycobacterium indicus pranii (MIP), a non-pathogenic Mycobacterium has been shown to exhibit cytotoxic effects on different cancer cells, including human cervical carcinoma cell line HeLa. However, the underlying mechanisms remain unknown. The aim of this study is to decipher the mechanism of MIP induced HeLa cell death. Methods: The cytotoxicity of Mycobacterium indicus pranii against HeLa cells was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Apoptosis was detected by annexin V and Propidium iodide (PI) staining. The assessment of reactive oxygen species (ROS) generation and cell cycle analysis were measured by flow cytometry. The expression of apoptosis associated genes was analyzed by real time PCR. Result: MIP could inhibit the proliferation of HeLa cell in a time and dose dependent manner but caused minor damage to normal cells. The induction of apoptosis was confirmed by the cell surface presentation of phosphatidyl serine, DNA fragmentation, and mitochondrial damage. MIP caused very early (as early as 30 minutes) transcriptional activation of p53, followed by a higher activation (32 fold) at 24 hours suggesting prime importance of p53 in MIP-induced apoptosis in HeLa cell. The up regulation of p53 dependent pro-apoptotic genes Bax, Bak, PUMA, and Noxa followed a lag phase that was required for the transcriptional p53 program. MIP also caused the transcriptional up regulation of Toll like receptor 2 and 4 after 30 minutes of MIP treatment suggesting recognition of MIP by toll like receptors. Moreover, MIP caused the inhibition of expression of HPV anti apoptotic gene E6, which is known to interfere with p53/PUMA/Bax apoptotic cascade. This inhibition might have played a role in transcriptional up regulation of PUMA and subsequently apoptosis. ROS was generated transiently which was concomitant with the highest transcription activation of p53 suggesting a plausible feedback loop network of p53 and ROS in the apoptosis of HeLa cells. Scavenger of ROS, such as N-acetyl-L-cysteine, decreased apoptosis suggesting ROS is an important effector of MIP induced apoptosis. Conclusion: Taken together, MIP possesses full potential to be a novel therapeutic agent in the clinical treatment of cervical cancer.

Keywords: cancer, mycobacterium, immunity, immunotherapy.

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