Search results for: small cell

Authors: R. Morbidelli, C. Saltalippi, A. Flammini, J. Dari

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This work analyses what effect systematic awareness-raising of the population on domestic water consumption produces. In a period where the availability of water is continually decreasing due to reduced rainfall, it is of paramount importance to raise awareness among the population. We conducted an experiment on a large sample of homes in urban areas of Central Italy. In the first phase, lasting three weeks, normal per capita, water consumption was quantified. Subsequently, instructions were given on how to save water during various uses in the household (showers, cleaning hands, use of water in toilets, watering small green areas, use of water in the kitchen, ...), and small visual messages were posted at water dispensers to remind users to behave properly. Finally, household consumption was assessed again for a further three weeks. This experiment made it possible to quantify the effect of the awareness-raising action on the reduction of water consumption without the use of any structural action (replacement of dispensers, improvement of the water system, ...).

Keywords: water saving, urban areas, awareness-raising, climate change

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Authors: Fardis Teifoori, Mehdi Dehghani, Idoia Postigo, Jorge Martinez

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Introduction: Many skin disorders, such as atopic dermatitis (AD), is characterized by inflammation, infection, and hyperplasia. In this work, keratinocytes from AD patients are used to study the pomegranate seed oil properties for skin care. Material and methods: Isolated keratinocytes from patients with AD were cultured and stimulated by IL-9 (20 ng/ml) and TNF-α (50ng/ml) for 48h to induce vascular endothelial growth factor (VEGF) and Regulated upon activation, normal T cell expressed and secreted (RANTES) production, respectively, in the presence of different concentrations of pomegranate seed oil (20, 50, 100, and 200 µM). Finally, the concentrations of RANTES and VEGF in the cell culture supernatant were quantified according to the standard protocol of commercial ELISA kits. Results: The results indicated that pomegranate seed oil concentrations of 50, 100, and 200 µM could significantly inhibit the production of VEGF and RANTES by stimulating keratinocytes with IL-9 (20 ng/ml) and TNF-α (50ng/ml), respectively. The decrease in VEGF and RANTES concentration in the presence of the pomegranate seed oil concentrations of 20 and 50 uM was not significant. Conclusion: It was concluded that pomegranate seed oil (PSO) counteracts atopic dermatitis conditions dose-dependently: with the highest effect at the concentration of 200 µM. We suggest that the inexpensive and easily available pomegranate seed oil is a good candidate for cosmetics and clinical utilization for skin care.

Keywords: atopic dermatitis, pomegranate, Punica granatum, RANTES, VEGF

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Authors: Thamo Sutharssan, Diogo Montalvao, Wen-Chung Wang, Yong Chen, Claudia Pisac

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A combined heat and power (CHP) system is an efficient and clean way to generate power (electricity). Heat produced by the CHP system can be used for water and space heating. The CHP system which uses hydrogen as fuel produces zero carbon emission. Its’ efficiency can reach more than 80% whereas that of a traditional power station can only reach up to 50% because much of the thermal energy is wasted. The other advantages of CHP systems include that they can decentralize energy generation, improve energy security and sustainability, and significantly reduce the energy cost to the users. This paper presents the economic benefits of using a CHP system in the domestic environment. For this analysis, natural gas is considered as potential fuel as the hydrogen fuel cell based CHP systems are rarely used. UK government incentives for CHP systems are also considered as the added benefit. Results show that CHP requires a significant initial investment in return it can reduce the annual energy bill significantly. Results show that an investment may be paid back in 7 years. After the back period, CHP can run for about 3 years as most of the CHP manufacturers provide 10-year warranty.

Keywords: combined heat and power, clean energy, hydrogen fuel cell, economic analysis of CHP, zero emission

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Authors: Min Sik Kim, Hwa Sung Shin

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From 2013, small eel farms, which are located in Han River Estuary, South Korea suffer damage because of unknown massive perish. In the middle of discussion that the cause of perish could be environmental changes or waste water, a large amount of unknown nemertean was discovered during that time. Some nemerteans are known releasing neurotoxin substance. In this study, we isolated intestinal bacteria using selective media and conducted 16s rDNA microbial identification by gene alignment. As a result, there was a type of bacteria producing TTX, blocks sodium-channel inducing organism’s death. TTX production from the bacteria was confirmed by ELISA and liquid chromatography coupled with mass spectrometer. Additionally, the activated-charcoal which has an ability to absorb small molecules like toxin was applied to fibrous mesh to prevent ingestion of aquatic organisms and increase applicable area. The viability of zebrafish in the water with TTX and charcoal fiber mat were not decreased meaning it could be used for solving the perishing problem in fish farm.

Keywords: nemertean, TTX, fiber mat, activated charcoal, zebrafish

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Authors: S. K. Ling, A. K. H. Kwan

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The production of SCC (self-consolidating concrete) generally requires a fairy high paste volume, ranging from 35% to 40% of the total concrete volume. Such high paste volume would lead to low dimensional stability and high carbon footprint. Direct lowering the paste volume would deteriorate the performance of SCC, especially the passing ability. It is often observed that at narrow gap of congested reinforcements, the paste often flows in the front leaving the coarse aggregate particle behind to block the subsequent flow of concrete. Herein, it is suggested to increase the mortar volume through incorporating ground sand with a mean size of 0.3 mm while keeping the paste volume small. Trial concrete mixes with paste volumes of 30% and 34% and different ground sand contents have been tested to demonstrate how the paste volume can be lowered without sacrificing the passing ability. Overall, the results demonstrated that the addition of ground sand would enable the achievement of high passing ability at a relatively small paste volume.

Keywords: ground sand, mortar volume, paste volume, self-consolidating concrete

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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: Danhua Xu, Zhenlan Qian, Zhu Wang, Jiayan Fu, Ling Wang

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A new business mode called rural complex is proposed by the China’s government to promote the development the economy in the rural area. However, for the sake of current national conditions including the great number of labor farmers owning the small scale farmlands and the uncertain enthusiasm from the enterprises, it is challenging to develop the big scale rural complex. To react to the dilemmas, this paper puts forward the micro-rural complex to boost the small scale farms by co-participation from a bottom-up mode. By analyzing the potential opportunities to find the suitable mode, exploring the interdisciplinary and interdepartmental co-participation way beyond architecture design and spatial planning between different actors, the paper tries to find a complete process towards the sustainable micro-rural complex and conducts an ongoing practice to optimize it, to bring new insights and reference to the rural development. According to the transformation of the economy, the micro-rural complex will develop into two phases, both of which can be discussed in three parts, the economic mode, the spatial support, and the Cooperating mechanism. The first stage is the agriculture co-participation based on the rise of Community supported agriculture (CSA) in which the consumers buy the products planted in an organic way from the farmers directly with a higher price to support the small-scale agriculture and overcome the food safety issues. The following stage sets up the agritourism catering the citizens with the restaurants, inns and other tourist service facilities to be planned and designed. In the whole process, the interdisciplinary co-participation will play an important role to provide the guidelines and consultation from the agronomists, architects and rural planners to the farmers. This mode has been applied to an on-going farm project, from which to explore the mode in a more practical way. In conclusion, the micro-rural complex aims at creating a balanced urban-rural relationship by co-participation taking advantage of the different actors. The spatial development is considered from the economic mode and social organization. The integration of the mode based on the small-scale agriculture will contribute to a sustainable growth and realize the long run development in the rural area.

Keywords: micro-rural complex, co-participation, sustainable development, China

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Authors: Wan-Ting Kuo, Yi-Wen Liu, Hsiao-Sheng Liu

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Annual incidence of bladder cancer increases in the world and occurs frequently in the male. Most common type is transitional cell carcinoma (TCC) which is treated by transurethral resection followed by intravesical administration of agents. In clinical treatment of bladder cancer, chemotherapeutic drugs-induced apoptosis is always used in patients. However, cancers usually develop resistance to chemotherapeutic drugs and often lead to aggressive tumors with worse clinical outcomes. Approximate 70% TCC recurs and 30% recurrent tumors progress to high-grade invasive tumors, indicating that new therapeutic agents are urgently needed to improve the successful rate of overall treatment. Nonapoptotic program cell death may assist to overcome worse clinical outcomes. Autophagy which is one of the nonapoptotic pathways provides another option for bladder cancer patients. Autophagy is reported as a potent anticancer therapy in some cancers. First of all, we established a mouse orthotopic bladder tumor formation model in order to create a similar tumor microenvironment. IVIS system and micro-ultrasound were utilized to noninvasively monitor tumor formation. In addition, we carried out intravesical treatment in our animal model to be consistent with human clinical treatment. In our study, we carried out intravesical instillation of the autophagy inducer in mouse orthotopic bladder tumor to observe tumor formation by noninvasive IVIS system and micro-ultrasound. Our results showed that bladder tumor formation is suppressed by the autophagy inducer, and there are no significant side effects in the physiology of mice. Furthermore, the autophagy inducer upregulated autophagy in bladder tissues of the treated mice was confirmed by Western blot, immunohistochemistry, and immunofluorescence. In conclusion, we reveal that a novel autophagy inducer with low side effects suppresses bladder tumor formation in our mouse orthotopic bladder tumor model, and it provides another therapeutic approach in bladder cancer patients.

Keywords: bladder cancer, transitional cell carcinoma, orthotopic bladder tumor formation model, autophagy

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Authors: Kyoung Lee

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Numerous aerobic bacteria have the ability to form multicellular communities on the surface layer of the air-liquid (A-L) interface as a biofilm called a pellicle. Pellicles occupied at the A-L interface will benefit from the utilization of oxygen from air and nutrient from liquid. Buoyancy of cells can be obtained by high surface tension at the A-L interface. Thus, formation of pellicles is an adaptive advantage in utilization of excess nutrients in the standing culture where oxygen depletion is easily set up due to rapid cell growth. In natural environments, pellicles are commonly observed on the surface of lake or pond contaminated with pollutants. Previously, we have shown that when cultured in standing LB media an alkylphenol-degrading bacteria Pseudomonas alkylphenolia KL28 forms pellicles in a diameter of 0.3-0.5 mm with a thickness of ca 40 µm. The pellicles have unique features for possessing flatness and unusual rigidity. In this study, the biogenesis of the circular pellicles has been investigated by observing the cell organization at early stages of pellicle formation and cell arrangements in pellicle, providing a clue for highly organized cellular arrangement to be adapted to the air-liquid niche. Here, we first monitored developmental patterns of pellicle from monolayer to multicellular organization. Pellicles were shaped by controlled growth of constituent cells which accumulate extracellular polymeric substance. The initial two-dimensional growth was transited to multilayers by a constraint force of accumulated self-produced extracellular polymeric substance. Experiments showed that pellicles are formed by clonal growth and even with knock-out of genes for flagella and pilus formation. In contrast, the mutants in the epm gene cluster for alginate-like polymer biosynthesis were incompetent in cell alignment for initial two-dimensional growth of pellicles. Electron microscopic and confocal laser scanning microscopic studies showed that the fully matured structures are highly packed by matrix-encased cells which have special arrangements. The cells on the surface of the pellicle lie relatively flat and inside longitudinally cross packed. HPLC analysis of the extrapolysaccharide (EPS) hydrolysate from the colonies from LB agar showed a composition with L-fucose, L-rhamnose, D-galactosamine, D-glucosamine, D-galactose, D-glucose, D-mannose. However, that from pellicles showed similar neutral and amino sugar profile but missing galactose. Furthermore, uronic acid analysis of EPS hydrolysates by HPLC showed that mannuronic acid was detected from pellicles not from colonies, indicating the epm-derived polymer is critical for pellicle formation as proved by the epm mutants. This study verified that for the circular pellicle architecture P. alkylphenolica KL28 cells utilized EPS building blocks different from that used for colony construction. These results indicate that P. alkylphenolica KL28 is a clever architect that dictates unique cell arrangements with selected EPS matrix material to construct sophisticated building, circular biofilm pellicles.

Keywords: biofilm, matrix, pellicle, pseudomonas

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Authors: Tanushree Jaitly, Shailendra Gupta, Leila Taher, Gerold Schuler, Julio Vera

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Genomics-based personalized medicine is a promising approach to fight aggressive tumors based on patient's specific tumor mutation and expression profiles. A remarkable case is, dendritic cell-based immunotherapy, in which tumor epitopes targeting patient's specific mutations are used to design a vaccine that helps in stimulating cytotoxic T cell mediated anticancer immunity. Here we present a computational pipeline for epitope-based personalized cancer vaccines using patient-specific haplotype and cancer mutation profiles. In the workflow proposed, we analyze Whole Exome Sequencing and RNA Sequencing patient data to detect patient-specific mutations and their expression level. Epitopes including the tumor mutations are computationally predicted using patient's haplotype and filtered based on their expression level, binding affinity, and immunogenicity. We calculate binding energy for each filtered major histocompatibility complex (MHC)-peptide complex using docking studies, and use this feature to select good epitope candidates further.

Keywords: cancer immunotherapy, epitope prediction, NGS data, personalized medicine

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Authors: Marek Halenár, Eva Tvrdá, Simona Baldovská, Ľubomír Ondruška, Peter Massányi, Adriana Kolesárová

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This study aimed to assess the in vitro effects of different concentrations of the Viscum album extract on the motility, viability, and reactive oxygen species (ROS) production by rabbit spermatozoa during different time periods (0, 2, and 8h). Spermatozoa motility was assessed by using the CASA (Computer aided sperm analysis) system. Cell viability was evaluated by using the metabolic activity MTT assay, and the luminol-based luminometry was applied to quantify the ROS formation. The CASA analysis revealed that low Viscum concentrations were able to prevent a rapid decline of spermatozoa motility, especially in the case of concentrations ranging between 1 and 5 µg/mL (P<0.05 with respect to time 8h). At the same time, concentrations ranging between 1 and 100 µg/mL of the extract led to a significant preservation of the cell viability (P<0.05 in case of 5, 50 and 100 µg/mL; P<0.01 with respect to 1 and 10 µg/mL, time 8h). 1 and 5 µg/mL of the extract exhibited antioxidant characteristics, translated into a significant reduction of the ROS production, particularly notable at time 8h (P<0.01). The results indicate that the Viscum extract is capable of delaying the damage inflicted to the spermatozoon by the in vitro environment.

Keywords: CASA, mistletoe, mitochondrial activity, motility, reactive oxygen species, rabbits, spermatozoa, Viscum album

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Authors: Guo-Hui Lu, Jing-Yi Cai, Ke-Yan Tan, Xiao-Cai Yin, Yu Zheng, Peng-Wei Shao, Yong-Liang Yang

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Soil erosion around abandoned mines is one of the important geological agents for pollutant diffuses to the lower reaches of the local river basin system. River loading of pollutants is an important parameter for remediation of abandoned mines. In order to obtain information on pollutant transport and diffusion downstream in mining area, the small tributary system of the Xiaxi River in Wanshan District of Guizhou Province was selected as the research area. Sediment and suspended matter samples were collected and determined for Pb, As, Hg, Zn, Co, Cd, Cu, Ni, Cr, and Mn by inductively coupled plasma mass spectrometry (ICP-MS) and atomic fluorescence spectrometry (AFS) with the pretreatment of wet digestion. Discussions are made for pollution status and spatial distribution characteristics. The total Hg content in the sediments ranged from 0.45 to 16.0 g/g (dry weight) with an average of 5.79 g/g, which was ten times higher than the limit of Class II soil for mercury by the National Soil Environmental Quality Standard. The maximum occurred at the intersection of the Jin River and the Xiaxi River. The potential ecological hazard index (RI) was used to evaluate the ecological risk of heavy metals in the sediments. The average RI value for the whole study area suggests the high potential ecological risk level. High Cd potential ecological risk was found at individual sites.

Keywords: heavy metal, risk assessment, sediment, suspended matter, Wanshan mercury mine, small tributary system

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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: Arunas Stirke, Neringa Bakute, Gatis Mozolevskis

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A technology of microfluidics is an emerging tool in the field of biology, medicine and chemistry. Microfluidic device is also known as ‘lab-on-a-chip’ technology [1]. In moving from macro- to microscale, there is unprecedented control over spatial and temporal gradients and patterns that cannot be captured in conventional Petri dishes and well plates [2]. However, there is not a single standard microfluidic chip designated for all purposes – every different field of studies needs a specific microchip with certain geometries, inlet/outlet, channel depth and other parameters to precisely regulate the required function. Since our group is studying an effect of pulsed electric field (PEF) to the cells, we have manufactured a microfluidic chip designated for high-throughput electroporation of cells. In our microchip, a cell culture chamber is divided into two parallel channels by a membrane, meanwhile electrodes for electroporation are attached to the wall of the channels. Both microchannels have their own inlet and outlet, enabling injection of transfection material separately. Our perspective is to perform electroporation of mammalian cells in two different ways: (1) plasmid and cells are injected in the same microchannel and (2) injected into separate microchannels. Moreover, oxygen and pH sensors are integrated on order to analyse cell viability parameters after PEF treatment.

Keywords: microfluidics, chip, fabrication, electroporation

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Authors: S. K. Thind, Aparjot Kaur

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Heat stress is one of the major environmental factors drastically reducing wheat production. Crop heat tolerance can be enhanced by preconditioning of plants by exogenous application of osmoprotectants. Presently, the effect of trehalose pretreatment (at 1 mM, and 1.5 nM) under heat stress of 35±2˚C (moderate) and 40±2˚ (severe) for four and eight hour was conducted in wheat (Tricticum aestivum L.) genotypes viz. HD2967, PBW 175, PBW 343, PBW 621, and PBW 590. Heat stress affects wide spectrum of physiological processes within plants that are irreversibly damaged by stress. Membrane thermal stability (MTS) and cell viability was significantly decreased under heat stress for eight hours. Pretreatment with trehalose improved MTS and cell viability under stress and this effect was more promotory with higher concentration. Thermal stability of photosynthetic apparatus differed markedly between genotypes and Hill reaction activity was recorded more in PBW621 followed by C306 as compared with others. In all genotypes photolysis of water showed decline with increase in temperature stress. Trehalose pretreatment helped in sustaining Hill reaction activity probably by stabilizing the photosynthetic apparatus against heat-induced photo inhibition. Both plant growth and development were affected by temperature in both shoot and root under heat stress. The reduction was compensated partially by trehalose (1.5 mM) application. Adaption to heat stress is associated with the metabolic adjustment which led to accumulation of soluble sugars including non-reducing and reducing for their role in adaptive mechanism. Higher acid invertase activity in shoot of tolerant genotypes appeared to be a characteristic for stress tolerance. As sucrose synthase play central role in sink strength and in studied wheat genotype was positively related to dry matter accumulation. The duration of heat stress for eight hours had more severe effect on these parameters and trehalose application at 1.5 mM ameliorated it to certain extent.

Keywords: heat stress, Triticum aestivum, trehalose, membrane thermal stability, triphenyl tetrazolium chloride, reduction test, growth, sugar metabolism

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Authors: Eitan Yefenof

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Glucocorticoid (GC) hormones, e.g. Dexamethasone and Prednisone, are widely used in the therapy of leukemia and lymphoma owing to their apoptogenic effect on lymphoid cells. However, the emergence of GC resistant cells during therapy is a major cause for treatment failure, urging the need for novel strategies that maintain leukemia sensitivity to the pro-apoptotic activity of GCs. GCs act by binding to the GC receptor (GR), which, in its inactive state, is sequestered in the cytosol by a multi-subunit complex of heat shock proteins. Upon ligand binding, the complex dissociates, allowing GR activation and translocation to the nucleus, where it regulates transcription of multiple genes. We demonstrated that in addition to gene expression, GR also regulates microRNA (miR) expression. Deep-sequencing analysis revealed 14 miRs that are regulated in GC-sensitive but resistant leukemias upon treatment with GC. GC up-regulates miR-103, miR-15~16 and miR-30e/d, while down-regulates miR-17, mir-18a, miR-19a, miR-19b, miR-20a and miR-92a (members of the miR-17∼92a multi-cistron). Upon transfection, miR-103 confers GC apoptotic sensitivity to otherwise GC-resistant cell. Furthermore, knocking down miR-103 expression reduces the GC apoptotic response of sensitive cells. miR-103 abrogates c-Myc expression, an oncogenic transcription factor which is deregulated in many cancers. In addition, miR-103 up-regulates Bim, a pro-apoptotic protein crucial for GC-induced death. Activated glycogen synthase kinase 3 (GSK3) is also crucial for GC-induced apoptosis. GSK3 is active in GC-sensitive but not in GC-resistant cells. We found that GSK3 associates with the GR multi-subunit complex. Upon GC exposure, it dissociates from the GR and interacts with Bim to enable activation of the mitochondrial apoptosis pathway. miR-103 mediated c-Myc ablation is followed by down-regulation of the multi-cistron miR-17~92a, in particular miR-18a and miR-20a. miR-18a targets GR for degradation whereas miR-20a targets Bim degradation. Hence, miR-103 acts, in concert with Bim and GR, as a "tumor suppressor" that leads to reduced proliferation, cell-cycle arrest and cell death. We suggest that miR-103 can provide a diagnostic tool that predicts the sensitivity of leukemia to GC based therapy. Furthermore, exosomal delivery of miR-103 or up-regulation of the endogenous miR-103 could confer apoptotic sensitivity to resistant cells at the outset, thus becoming a useful therapeutic tool combined with GCs.

Keywords: apoptosis, leukemia, micro-RNA, steroids

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Authors: Aoxue Yang, Weili Tian, Yonghe Wu, Haikun Liu

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Brain tumor stem cells (BTSCs) are resistant to therapy and give rise to recurrent tumors. These rare and elusive cells are likely to disseminate during cancer progression, and some may enter dormancy, remaining viable but not increasing. The identification of dormant BTSCs is thus necessary to design effective therapies for glioblastoma (GBM) patients. Little progress has been made in therapeutic treatment of glioblastoma in the last decade despite rapid progress in molecular understanding of brain tumors1. Here we show that the stress hormone glucocorticoid is essential for the maintenance of brain tumor stem cells (BTSCs), which are resistant to conventional therapy. The glucocorticoid receptor (GR) regulates metabolic plasticity and chemoresistance of the dormant BTSC via controlling expression of GPD1 (glycerol-3-phosphate dehydrogenase 1), which is an essential regulator of lipid metabolism in BTSCs. Genomic, lipidomic and cellular analysis confirm that GR/GPD1 regulation is essential for BTSCs metabolic plasticity and survival. We further demonstrate that the GR agonist dexamethasone (DEXA), which is commonly used to control edema in glioblastoma, abolishes the effect of chemotherapy drug temozolomide (TMZ) by upregulating GPD1 and thus promoting tumor cell dormancy in vivo, this provides a mechanistic explanation and thus settle the long-standing debate of usage of steroid in brain tumor patient edema control. Pharmacological inhibition of GR/GPD1 pathway disrupts metabolic plasticity of BTSCs and prolong animal survival, which is superior to standard chemotherapy. Patient case study shows that GR antagonist mifepristone blocks tumor progression and leads to symptomatic improvement. This study identifies an important mechanism regulating cancer stem cell dormancy and provides a new opportunity for glioblastoma treatment.

Keywords: cancer stem cell, dormancy, glioblastoma, glycerol-3-phosphate dehydrogenase 1, glucocorticoid receptor, dexamethasone, RNA-sequencing, phosphoglycerides.

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Authors: Eva Tvrdá, Hana Greifová, Peter Ivanič, Norbert Lukáč

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The aim of this study was to evaluate the dose- and time-dependent in vitro effects of berberine (BER), a natural alkaloid with numerous biological properties on bovine spermatozoa during three time periods (0 h, 2 h, 24 h). Bovine semen samples were diluted and cultivated in physiological saline solution containing 0.5% DMSO together with 200, 100, 50, 10, 5, and 1 μmol/L BER. Spermatozoa motility was assessed using the computer assisted semen analyzer. The viability of spermatozoa was assessed by the metabolic (MTT) assay, production of superoxide radicals was quantified using the nitroblue tetrazolium (NBT) test, and chemiluminescence was used to evaluate the generation of reactive oxygen species (ROS). Cell lysates were prepared and the extent of lipid peroxidation (LPO) was evaluated using the TBARS assay. The results of the movement activity showed a significant increase in the motility during long term cultivation in case of concentrations ranging between 1 and 10 μmol/L BER (P < 0.01; P < 0.001; 24 h). At the same time, supplementation of 1, 5 and 10 μmol/L BER led to a significant preservation of the cell viability (P < 0.001; 24 h). BER addition at a range of 1-50 μmol/L also provided a significantly higher protection against superoxide (P < 0.05) and ROS (P < 0.001; P < 0.01) overgeneration as well as LPO (P < 0.01; P<0.05) after a 24 h cultivation. We may suggest that supplementation of BER to bovine spermatozoa, particularly at concentrations ranging between 1 and 50 μmol/L, may offer protection to the motility, viability and oxidative status of the spermatozoa, particularly notable at 24 h.

Keywords: berberine, bulls, motility, oxidative profile, spermatozoa, viability

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Authors: Jayarama Reddy, Anand S., H., Sundaram, Jeldi Hemachandran

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Forty two strains of Trichoderma sp. were isolated from cultivated lands around Bangalore and analyzed for their antagonistic potential against Macrophomina phaseolina. The potential of biocontrol agents ultimately lies in their capacity to control pathogens in vivo. Bioefficacy studies were hence conducted using chickpea (Cicer arientum c.v. Annigeri) as an experimental plant by the roll paper towel method. Overall the isolates T6, T35, T30, and T25 showed better antagonistic potential in addition to enhancing plant growth. The production of chitinases to break down the mycelial cell walls of fungal plant pathogens has been implicated as a major cause of biocontrol activity. In order to study the mechanism of biocontrol against Macrophomina phaseolina, ten better performing strains were plated on media, amended with colloidal chitin and Sclerotium rolfsii cell wall extract. All the isolates showed chitinolytic activity on day three as well as day five. Production of endochitinase and exochitinase were assayed in liquid media using colloidal chitin amended broth. Strains T35 and T6 displayed maximum endochitinase and exochitinase activity. Although all strains exhibited cellulase activity, the quantum of enzyme produced was higher in T35 and T6. The results also indicate a positive correlation between enzyme production and bioefficacy.

Keywords: biocontrol, bioefficacy, cellulase, chitinase

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Authors: Xinyong Li

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A new coupling system was constructed by combining photo-electrochemical cell with eletro-fenton cell (PEC-EF). The electrode material in this system was derived from MnyFe₁₋yCo Prussian-Blue-Analog (PBA). Mn₀.₄Fe₀.₆Co₀.₆₇-N@C spin-coated on carbon paper behaved as the gas diffusion cathode and Mn₀.₄Fe₀.₆Co₀.₆₇O₂.₂ spin-coated on fluorine-tin oxide glass (FTO) as anode. The two separated cells could degrade Sulfamethoxazole (SMX) simultaneously and some coupling mechanisms by PEC and EF enhancing the degradation efficiency were investigated. The continuous on-site generation of H₂O₂ at cathode through an oxygen reduction reaction (ORR) was realized over rotating ring-disk electrode (RRDE). The electron transfer number (n) of the ORR with Mn₀.₄Fe₀.₆Co₀.₆₇-N@C was 2.5 in the selected potential and pH range. The photo-electrochemical properties of Mn₀.₄Fe₀.₆Co₀.₆₇O₂.₂ were systematically studied, which displayed good response towards visible light. The photo-induced electrons at anode can transfer to cathode for further use. Efficient photo-electro-catalytic performance was observed in degrading SMX. Almost 100% SMX removal was achieved in 120 min. This work not only provided a highly effective technique for antibiotic treatment but also revealed the synergic effect between PEC and EF.

Keywords: Electro-Fenton, photo-electrochemical, synergic effect, sulfamethoxazole

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Authors: Vilas Warudkar, Dinkar Janghel, Siraj Ahmed

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Rapid development experienced by India requires huge amount of energy. Actual supply capacity additions have been consistently lower than the targets set by the government. According to World Bank 40% of residences are without electricity. In 12th five year plan 30 GW grid interactive renewable capacity is planned in which 17 GW is Wind, 10 GW is from solar and 2.1 GW from small hydro project, and rest is compensated by bio gas. Renewable energy (RE) and energy efficiency (EE) meet not only the environmental and energy security objectives, but also can play a crucial role in reducing chronic power shortages. In remote areas or areas with a weak grid, wind energy can be used for charging batteries or can be combined with a diesel engine to save fuel whenever wind is available. India according to IEC 61400-1 belongs to class IV Wind Condition; it is not possible to set up wind turbine in large scale at every place. So, the best choice is to go for small scale wind turbine at lower height which will have good annual energy production (AEP). Based on the wind characteristic available at MANIT Bhopal, rotor for small scale wind turbine is designed. Various Aero foil data is reviewed for selection of airfoil in the Blade Profile. Airfoil suited of Low wind conditions i.e. at low Reynold’s number is selected based on Coefficient of Lift, Drag and angle of attack. For designing of the rotor blade, standard Blade Element Momentum (BEM) Theory is implanted. Performance of the Blade is estimated using BEM theory in which axial induction factor and angular induction factor is optimized using iterative technique. Rotor performance is estimated for particular designed blade specifically for low wind Conditions. Power production of rotor is determined at different wind speeds for particular pitch angle of the blade. At pitch 15o and velocity 5 m/sec gives good cut in speed of 2 m/sec and power produced is around 350 Watts. Tip speed of the Blade is considered as 6.5 for which Coefficient of Performance of the rotor is calculated 0.35, which is good acceptable value for Small scale Wind turbine. Simple Load Model (SLM, IEC 61400-2) is also discussed to improve the structural strength of the rotor. In SLM, Edge wise Moment and Flap Wise moment is considered which cause bending stress at the root of the blade. Various Load case mentioned in the IEC 61400-2 is calculated and checked for the partial safety factor of the wind turbine blade.

Keywords: annual energy production, Blade Element Momentum Theory, low wind Conditions, selection of airfoil

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Authors: Peerawat Khongkliang, Prawit Kongjan, Tsuyoshi Imai, Poonsuk Prasertsan, Sompong O-Thong

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A two-stage thermophilic fermentation and electrohydrogenesis process was used to convert cassava starch processing wastewater into hydrogen gas. Maximum hydrogen yield from fermentation stage by Thermoanaerobacterium thermosaccharolyticum PSU-2 was 248 mL H2/g-COD at optimal pH of 6.5. Optimum hydrogen production rate of 820 mL/L/d and yield of 200 mL/g COD was obtained at HRT of 2 days in fermentation stage. Cassava starch processing wastewater fermentation effluent consisted of acetic acid, butyric acid and propionic acid. The effluent from fermentation stage was used as feedstock to generate hydrogen production by microbial electrolysis cell (MECs) at an applied voltage of 0.6 V in second stage with additional 657 mL H2/g-COD was produced. Energy efficiencies based on electricity needed for the MEC were 330 % with COD removals of 95 %. The overall hydrogen yield was 800-900 mL H2/g-COD. Microbial community analysis of electrohydrogenesis by DGGE shows that exoelectrogens belong to Acidiphilium sp., Geobacter sulfurreducens and Thermincola sp. were dominated at anode. These results show two-stage thermophilic fermentation, and electrohydrogenesis process improved hydrogen production performance with high hydrogen yields, high gas production rates and high COD removal efficiency.

Keywords: cassava starch processing wastewater, biohydrogen, thermophilic fermentation, microbial electrolysis cell

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Authors: Dunwen Zuo, Yongfang Deng, Bo Song

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An attempt is made here to measure the forces of three directions, under conditions of different feed speeds, different tilt angles of tool and without or with the pin on the tool, by using octagonal ring dynamometer in the AA2024 aluminum FSJ (Friction Stir Joining) process, and investigate how four main factors influence forces in the FSJ process. It is found that, high feed speed lead to small feed force and small lateral force, but high feed speed leads to large feed force in the stable joining stage of process. As the rotational speed increasing, the time of axial force drop from the maximum to the minimum required increased in the push-up process. In the stable joining stage, the rotational speed has little effect on the feed force; large rotational speed leads to small lateral force and axial force. The maximum axial force increases as the tilt angle of tool increases at the downward movement stage. At the moment of start feeding, as tilt angle of tool increases, the amplitudes of the axial force increasing become large. In the stable joining stage, with the increase of tilt angle of tool, the axial force is increased, the lateral force is decreased, and the feed force almost unchanged. The tool with pin will decrease axial force in the downward movement stage. The feed force and lateral force will increase, but the axial force will reduced in the stable joining stage by using the tool with pin compare to by using the tool without pin.

Keywords: FSJ, force factor, AA2024 aluminum, friction stir joining

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Authors: Stéphanie Looser, Walter Wehrmeyer

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Given the limited research on Small and Mediumsized Enterprises’ (SMEs) contribution to Corporate Social Responsibility (CSR) and even scarcer research on Swiss SMEs, this paper helps to fill these gaps by enabling the identification of supranational SME parameters and to make a contribution to the evolving field of these topics. Thus, the paper investigates the current state of SME practices in Switzerland and across 15 other countries. Combining the degree to which SMEs demonstrate an explicit (or business case) approach or see CSR as an implicit moral activity with the assessment of their attributes for “variety of capitalism” defines the framework of this comparative analysis. According to previous studies, liberal market economies, e.g. in the United States (US) or United Kingdom (UK), are aligned with extrinsic CSR, while coordinated market systems (in Central European or Asian countries) evolve implicit CSR agendas. To outline Swiss small business CSR patterns in particular, 40 SME owner-managers were interviewed. The transcribed interviews were coded utilising MAXQDA for qualitative content analysis. A secondary data analysis of results from different countries (i.e., Australia, Austria, Chile, Cameroon, Catalonia (notably a part of Spain that seeks autonomy), China, Finland, Germany, Hong Kong (a special administrative region of China), Italy, Netherlands, Singapore, Spain, Taiwan, UK, US) lays groundwork for this comparative study on small business CSR. Applying the same coding categories (in MAXQDA) for the interview analysis as well as for the secondary data research while following grounded theory rules to refine and keep track of ideas generated testable hypotheses and comparative power on implicit (and the lower likelihood of explicit) CSR in SMEs retrospectively. The paper identifies Swiss small business CSR as deep, profound, “soul”, and an implicit part of the day-to-day business. Similar to most Central European, Mediterranean, Nordic, and Asian countries, explicit CSR is still very rare in Swiss SMEs. Astonishingly, also UK and US SMEs follow this pattern in spite of their strong and distinct liberal market economies. Though other findings show that nationality matters this research concludes that SME culture and its informal CSR agenda are strongly formative and superseding even forces of market economies, nationally cultural patterns, and language. In a world of “big business”, explicit “business case” CSR, and the mantra that “CSR must pay”, this study points to a distinctly implicit small business CSR model built on trust, physical closeness, and virtues that is largely detached from the bottom line. This pattern holds for different cultural contexts and it is concluded that SME culture is stronger than nationality leading to a supra-national, monolithic SME CSR approach. Hence, classifications of countries by their market system or capitalism, as found in the comparative capitalism literature, do not match the CSR practices in SMEs as they do not mirror the peculiarities of their business. This raises questions on the universality and generalisability of management concepts.

Keywords: CSR, comparative study, cultures of capitalism, small, medium-sized enterprises

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Authors: Sanat K. Dash, Rama S. Verma, Sarit K. Das

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A logarithmic microfluidic shear device was designed and fabricated for cellular mechanotransduction studies. The device contains four cell culture chambers in which flow was modulated to achieve a logarithmic increment. Resistance values were optimized to make the device compact. The network of resistances was developed according to a unique combination of series and parallel resistances as found via optimization. Simulation results done in Ansys 16.1 matched the analytical calculations and showed the shear stress distribution at different inlet flow rates. Fabrication of the device was carried out using conventional photolithography and PDMS soft lithography. Flow profile was validated taking DI water as working fluid and measuring the volume collected at all four outlets. Volumes collected at the outlets were in accordance with the simulation results at inlet flow rates ranging from 1 ml/min to 0.1 ml/min. The device can exert fluid shear stresses ranging four orders of magnitude on the culture chamber walls which will cover shear stress values from interstitial flow to blood flow. This will allow studying cell behavior in the long physiological range of shear stress in a single run reducing number of experiments.

Keywords: microfluidics, mechanotransduction, fluid shear stress, physiological shear

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Authors: Grigoria Athanasaki, Veerarajan Vimala, A. M. Kannan, Louis Cindrella

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Energy usage has been increased throughout the years, leading to severe environmental impacts. Since the majority of the energy is currently produced from fossil fuels, there is a global need for clean energy solutions. Proton Exchange Membrane Fuel Cells (PEMFCs) offer a very promising solution for transportation applications because of their solid configuration and low temperature operations, which allows them to start quickly. One of the main components of PEMFCs is the Gas Diffusion Layer (GDL), which manages water and gas transport and shows direct influence on the fuel cell performance. In this work, a novel dual-layer GDL with gradient porosity was prepared, using polyethylene glycol (PEG) as pore former, to improve the gas diffusion and water management in the system. The microporous layer (MPL) of the fabricated GDL consists of carbon powder PUREBLACK, sodium dodecyl sulfate as a surfactant, 34% wt. PTFE and the gradient porosity was created by applying one layer using 30% wt. PEG on the carbon substrate, followed by a second layer without using any pore former. The total carbon loading of the microporous layer is ~ 3 mg.cm-2. For the assembly of the catalyst layer, Nafion membrane (Ion Power, Nafion Membrane NR211) and Pt/C electrocatalyst (46.1% wt.) were used. The catalyst ink was deposited on the membrane via microspraying technique. The Pt loading is ~ 0.4 mg.cm-2, and the active area is 5 cm2. The sample was ex-situ characterized via wetting angle measurement, Scanning Electron Microscopy (SEM), and Pore Size Distribution (PSD) to evaluate its characteristics. Furthermore, for the performance evaluation in-situ characterization via Fuel Cell Testing using H2/O2 and H2/air as reactants, under 50, 60, 80, and 100% relative humidity (RH), took place. The results were compared to a single layer GDL, fabricated with the same carbon powder and loading as the dual layer GDL, and a commercially available GDL with MPL (AvCarb2120). The findings reveal high hydrophobic properties of the microporous layer of the GDL for both PUREBLACK based samples, while the commercial GDL demonstrates hydrophilic behavior. The dual layer GDL shows high and stable fuel cell performance under all the RH conditions, whereas the single layer manifests a drop in performance at high RH in both oxygen and air, caused by catalyst flooding. The commercial GDL shows very low and unstable performance, possibly because of its hydrophilic character and thinner microporous layer. In conclusion, the dual layer GDL with PEG appears to have improved gas diffusion and water management in the fuel cell system. Due to its increasing porosity from the catalyst layer to the carbon substrate, it allows easier access of the reactant gases from the flow channels to the catalyst layer, and more efficient water removal from the catalyst layer, leading to higher performance and stability.

Keywords: gas diffusion layer, microporous layer, proton exchange membrane fuel cells, relative humidity

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Authors: Shabnam Abdi, Behzad Toosi

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Background: Melanoma is the most lethal type of malignant skin cancer in humans and dogs since it spreads rapidly throughout the body. Despite significant advances in treatment, cancer at an advanced stage has a poor prognosis. Hence, more effective treatments are needed to enhance outcomes with fewer side effects. Erythropoietin-producing hepatocellular receptors are the largest family of receptor tyrosine kinases and are divided into two subfamilies, EphA and EphB, both of which play a significant role in disease, especially cancer. Due to their association with proliferation and invasion in many aggressive types of cancer, Eph receptor tyrosine kinases (Eph RTKs) are promising cancer therapy molecules. Because these receptors have not been studied in canine melanoma, we investigated how EphA2 influences survival and tumorigenicity of melanoma cells. Methods: Expression of EphA2 protein in canine melanoma cell lines and human melanoma cell line was evaluated by Western blot. Melanoma cells were transduced with lentiviral particles encoding Eph-targeting shRNAs or non-silencing shRNAs (control) for silencing the expression of EphA2 receptor, and silencing was confirmed by Western blotting and immunofluorescence. The effect of siRNA treatment on cellular proliferation, colony formation, tumorsphere assay, invasion was analyzed by Resazurin assay Matrigel invasion assay, respectively. Results: Expression of EphA2 was detected in canine and human melanoma cell lines. Moreover, stably silencing EphA2 by specific shRNAs significantly and consistently decreased the expression of EphA2 protein in both human and canine melanoma cells. Proliferation, colony formation, tumorsphere and invasion of melanoma cells were significantly decreased in EphA2 siRNA-treated cells compared to control. Conclusion: Our data provide the first functional evidence that the EphA2 receptor plays a critical role in the malignant cellular behavior of melanoma in both human and dogs.

Keywords: ephA2, targeting, melanoma, human, canine

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Authors: James Banda, Jupiter Simbeye, Essau Chisale, Geoffrey Kanyerere, Kings Kamtambe

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Improved solar tent dryers for processing small fish species were designed to reduce post-harvest fish losses and improve supply of quality fish products in the southern part of Lake Malawi under CultiAF project. A comparative analysis of the quality of Diplotaxodon (Ndunduma) from Lake Malawi processed in solar tent dryer and open sun drying was conducted using proximate analysis, microbial analysis and sensory evaluation. Proximates for solar tent dried fish and open sun dried fish in terms of proteins, fats, moisture and ash were 63.3±0.15% and 63.3±0.34%, 19.6±0.09% and 19.9±0.25%, 8.3±0.12% and 17.0±0.01%, and 15.6±0.61% and 21.9±0.91% respectively. Crude protein and crude fat showed non-significant differences (p = 0.05), while moisture and ash content were significantly different (p = 001). Open sun dried fish had significantly higher numbers of viable bacteria counts (5.2×10⁶ CFU) than solar tent dried fish (3.9×10² CFU). Most isolated bacteria from solar tent dried and open sun dried fish were 1.0×10¹ and 7.2×10³ for Total coliform, 0 and 4.5 × 10³ for Escherishia coli, 0 and 7.5 × 10³ for Salmonella, 0 and 5.7×10² for shigella, 4.0×10¹ and 6.1×10³ for Staphylococcus, 1.0×10¹ and 7.0×10² for vibrio. Qualitative evaluation of sensory properties showed higher acceptability of 3.8 for solar tent dried fish than 1.7 for open sun dried fish. It is concluded that promotion of solar tent drying in processing small fish species in Malawi would support small-scale fish processors to produce quality fish in terms of nutritive value, reduced microbial contamination, sensory acceptability and reduced moisture content.

Keywords: diplotaxodon, Malawi, open sun drying, solar tent drying

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Authors: T. Ferreira, A. Kulkarni, C. Bretscher, K. Richter, M. Ehrlich, A. Marchini

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H-1 protoparvovirus (H-1PV) is a virus with inherent oncolytic and oncosuppressive activities while remaining non-pathogenic in humans. H-1PV was the first oncolytic parvovirus to undergo clinical testing. Results from trials in patients with glioblastoma or pancreatic carcinoma showed an excellent safety profile and first signs of efficacy. H-1PV infection is vastly dependent on cellular factors, from cell attachment and entry to viral replication and egress. Hence, we believe that the characterisation of the parvovirus life cycle would ultimately help further improve H-1PV clinical outcome. In the present study, we explored the entry pathway of H-1PV in cervical HeLa and glioma NCH125 cancer cell lines. Electron and confocal microscopy showed viral particles associated with clathrin-coated pits and vesicles, providing the first evidence that H-1PV cell entry occurs through clathrin-mediated endocytosis. Accordingly, we observed that by blocking clathrin-mediated endocytosis with hypertonic sucrose, chlorpromazine, or pitstop 2, H-1PV transduction was markedly decreased. Accordingly, siRNA-mediated knockdown of AP2M1, which retains a crucial role in clathrin-mediated endocytosis, verified the reliance of H-1PV on this route to enter HeLa and NCH125 cancer cells. By contrast, we found no evidence of viral entry through caveolae-mediated endocytosis. Indeed, pre-treatment of cells with nystatin or methyl-β-cyclodextrin, both inhibitors of caveolae-mediated endocytosis, did not affect viral transduction levels. Unexpectedly, siRNA-mediated knockdown of caveolin-1, the main driver of caveolae-mediated endocytosis, increased H-1PV transduction, suggesting caveolin-1 is a negative modulator of H-1PV infection. We also show that H-1PV entry is dependent on dynamin, a protein responsible for mediating the scission of vesicle neck and promoting further internalisation. Furthermore, since dynamin inhibition almost completely abolished H-1PV infection, makes it unlikely that H-1PV uses macropinocytosis as an alternative pathway to enter cells. After viral internalisation, H-1PV passes through early to late endosomes as observed by confocal microscopy. Inside these endocytic compartments, the acidic environment proved to be crucial for a productive infection. Inhibition of acidification of pH dramatically reduced H-1PV transduction. Besides, a fraction of H-1PV particles was observed inside LAMP1-positive lysosomes, most likely following a non-infectious route. To the author's best knowledge, this is the first study to characterise the cell entry pathways of H-1PV. Along these lines, this work will further contribute to understand H-1PV oncolytic properties as well as to improve its clinical potential in cancer virotherapy.

Keywords: clathrin-mediated endocytosis, H-1 parvovirus, oncolytic virus, virus entry

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Authors: B. Gonzalez-Yebra, H. Barajas, P. Palomares, M. Hernandez, O. Torres, M. Ayala, A. L. González, G. Vazquez-Ortiz, M. L. Guzman

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Leukemia arises by molecular alterations of the normal hematopoietic stem cell (HSC) transforming it into a leukemic stem cell (LSC) with high cell proliferation, self-renewal, and cell differentiation. Chronic myeloid leukemia (CML) originates from an LSC-leading to elevated proliferation of myeloid cells and acute lymphoblastic leukemia (ALL) originates from an LSC development leading to elevated proliferation of lymphoid cells. In both cases, LSC can be identified by multicolor flow cytometry using several antibodies. However, to date, LSC levels in peripheral blood (PB) are not established well enough in ALL and CML patients. On the other hand, the detection of the minimal residue disease (MRD) in leukemia is mainly based on the identification of the mRNA bcr-abl gene in CML patients and some other genes in ALL patients. There is no a properly biomarker to detect MDR in both types of leukemia. The objective of this study was to determine mRNA bcr-abl and the percentage of LSC in peripheral blood of patients with CML and ALL and identify a possible association between the amount of LSC in PB and clinical data. We included in this study 19 patients with Leukemia. A PB sample was collected per patient and leukocytes were obtained by Ficoll gradient. The immunophenotype for LSC CD34+ was done by flow cytometry analysis with CD33, CD2, CD14, CD16, CD64, HLA-DR, CD13, CD15, CD19, CD10, CD20, CD34, CD38, CD71, CD90, CD117, CD123 monoclonal antibodies. In addition, to identify the presence of the mRNA bcr-abl by RT-PCR, the RNA was isolated using TRIZOL reagent. Molecular (presence of mRNA bcr-abl and LSC CD34+) and clinical results were analyzed with descriptive statistics and a multiple regression analysis was performed to determine statistically significant association. In total, 19 patients (8 patients with ALL and 11 patients with CML) were analyzed, 9 patients with de novo leukemia (ALL = 6 and CML = 3) and 10 under treatment (ALL = 5 and CML = 5). The overall frequency of mRNA bcr-abl was 31% (6/19), and it was negative in ALL patients and positive in 80% in CML patients. On the other hand, LSC was determined in 16/19 leukemia patients (%LSC= 0.02-17.3). The Novo patients had higher percentage of LSC (0.26 to 17.3%) than patients under treatment (0 to 5.93%). The amount of LSC was significantly associated with the amount of LSC were: absence of treatment, the absence of splenomegaly, and a lower number of leukocytes, negative association for the clinical variables age, sex, blasts, and mRNA bcr-abl. In conclusion, patients with de novo leukemia had a higher percentage of circulating LSC than patients under treatment, and it was associated with clinical parameters as lack of treatment, absence of splenomegaly and a lower number of leukocytes. The mRNA bcr-abl detection was only possible in the series of patients with CML, and molecular detection of LSC could be identified in the peripheral blood of all leukemia patients, we believe the identification of circulating LSC may be used as biomarker for the detection of the MRD in leukemia patients.

Keywords: stem cells, leukemia, biomarkers, flow cytometry

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