Search results for: cell cultivation

Authors: Paul Hernandez-Herrera, Fernando Montoya, Juan Manuel Rendon, Alberto Darszon, Gabriel Corkidi

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Intracellular calcium ([Ca2+]i) regulates sperm motility. The analysis of [Ca2+]i has been traditionally achieved in two dimensions while the real movement of the cell takes place in three spatial dimensions. Due to optical limitations (high speed cell movement and low light emission) important data concerning the three dimensional movement of these flagellated cells had been neglected. Visualizing [Ca2+]i in 3D is not a simple matter since it requires complex fluorescence microscopy techniques where the resulting images have very low intensity and consequently low SNR (Signal to Noise Ratio). In 4D sequences, this problem is magnified since the flagellum oscillates (for human sperm) at least at an average frequency of 15 Hz. In this paper, a novel approach to extract the flagellum’s center-line in 4D stacks is presented. For this purpose, an iterative algorithm based on the fast-marching method is proposed to extract the flagellum’s center-line. Quantitative and qualitative results are presented in a 4D stack to demonstrate the ability of the proposed algorithm to trace the flagellum’s center-line. The method reached a precision and recall of 0.96 as compared with a semi-manual method.

Keywords: flagellum, minimal path, segmentation, sperm

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Authors: Ciprian Briciu-Burghina, Brendan Heery, Dermot Brabazon, Fiona Regan

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Good quality bathing water is a highly desirable natural resource which can provide major economic, social, and environmental benefits. Both in Ireland and Europe, such water bodies are managed under the European Directive for the management of bathing water quality (BWD). The BWD aims mainly: (i) to improve health protection for bathers by introducing stricter standards for faecal pollution assessment (E. coli, enterococci), (ii) to establish a more pro-active approach to the assessment of possible pollution risks and the management of bathing waters, and (iii) to increase public involvement and dissemination of information to the general public. Standard methods for E. coli and enterococci quantification rely on cultivation of the target organism which requires long incubation periods (from 18h to a few days). This is not ideal when immediate action is required for risk mitigation. Municipalities that oversee the bathing water quality and deploy appropriate signage have to wait for laboratory results. During this time, bathers can be exposed to pollution events and health risks. Although forecasting tools exist, they are site specific and as consequence extensive historical data is required to be effective. Another approach for early detection of faecal pollution is the use of marker enzymes. β-glucuronidase (GUS) is a widely accepted biomarker for E. coli detection in microbiological water quality control. GUS assay is particularly attractive as they are rapid, less than 4 h, easy to perform and they do not require specialised training. A method for on-site detection of GUS from environmental samples in less than 75 min was previously demonstrated. In this study, the capability of ColiSense as an early warning system for faecal pollution in freshwater is assessed. The system successfully detected GUS activity in all of the 45 freshwater samples tested. GUS activity was found to correlate linearly with E. coli (r2=0.53, N=45, p < 0.001) and enterococci (r2=0.66, N=45, p < 0.001) Although GUS is a marker for E. coli, a better correlation was obtained for enterococci. For this study water samples were collected from 5 rivers in the Dublin area over 1 month. This suggests a high diversity of pollution sources (agricultural, industrial, etc) as well as point and diffuse pollution sources were captured in the sample size. Such variety in the source of E. coli can account for different GUS activities/culturable cell and different ratios of viable but not culturable to viable culturable bacteria. A previously developed protocol for the recovery and detection of E. coli was coupled with a miniaturised fluorometer (ColiSense) and the system was assessed for the rapid detection FIB in freshwater samples. Further work will be carried out to evaluate the system’s performance on seawater samples.

Keywords: faecal pollution, β-glucuronidase (GUS), bathing water, E. coli

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Authors: Christopher R. Triggle, Hong Ding, Khaled Machaca, Gnanapragasam Arunachalam

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The antidiabetic drug, metformin, has been in clinical use for over 50 years and remains the first choice drug for the treatment of type two diabetes. In addition to its effectiveness as an oral anti-hyperglycaemic drug metformin also possesses vasculoprotective effects that are assumed to be secondary to its ability to reduce insulin resistance and control glycated hemoglobin levels; however, recent data from our laboratory indicate that metformin also has direct vasoprotective effects that are mediated, at least in part, via the anti-ageing gene, SIRT1. Diabetes is a major risk factor for the development of cardiovascular disease (CVD) and it is also well established that tobacco use further enhances the risk of CVD; however, it is not known whether treatment with metformin can offset the negative effects of diabetes and tobacco use on cardiac function. The current study was therefore designed to investigate 1: the effects of hyperglycaemia (HG) either alone or in the presence of elevated fatty acids (palmitate) and nicotine on the protein expression levels of the deacetylase sirtuin 1 (the protein product of SIRT1), anti-apoptotic Bcl-2, pro-apoptotic BIM and the pro-apoptotic, tumour suppressor protein, acetylated p53 in cardiomyocytes. 2: the ability of metformin to prevent the detrimental effects of HG, palmitate and nicotine on cardiomyocyte survival. Cell culture protocols were designed using a rat cardiomyocyte cell line, H9c2, either under normal glycaemic (NG) conditions of 5.5mM glucose, or hyperglycaemic conditions (HG) of 25mM glucose with, or without, added palmitate (250μM) or nicotine (1.0mM) for 24h. Immuno-blotting was used to detect the expression of sirtuin 1, Bcl-2, BIM, acetylated (Ac)-p53, p53 with β-actin used as the reference protein. Exposure to HG, palmitate, or nicotine alone significantly reduced expression of sirtuin1, Bcl-2 and raised the expression levels of acetylated p53 and BIM; however, the combination of HG, palmitate and nicotine had a synergistic effect to significantly suppress the expression levels of sirtuin 1 and Bcl-2, but further enhanced the expression of Ac-p53, and BIM. The inclusion of 1000μM, but not 50μM, metformin in the H9c2 cell culture protocol prevented the effects of HG, palmitate and nicotine on the pro-apoptotic pathways. Collectively these data indicate that metformin, in addition to its anti-hyperglycaemic and vasculoprotective properties, also has direct cardioprotective actions that offset the negative effects of hyerglycaemia, elevated free fatty acids and nicotine on cardiac cell survival. These data are of particular significance for the treatment of patients with diabetes who are also smokers as the inclusion of metformin in their therapeutic treatment plan should help reduce cardiac-related morbidity and mortality.

Keywords: apoptosis, cardiac muscle, diabetes, metformin, nicotine

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Authors: Saman Khan, Imrana Naseem

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Copper is an essential trace element required for pro-angiogenic co-factors including vascular endothelial growth factor (VEGF). Elevated levels of copper are found in various types of cancer including prostrate, colon, breast, lung and liver for angiogensis and metastasis. Therefore, targeting copper via copper-specific chelators in cancer cells can be developed as effective anticancer treatment strategy. In continuation of our pursuit to design and synthesize copper chelators, herein we opted for a reaction to incorporate di-(2-picolyl) amine in 3-(bromoacetyl) coumarin (parent backbone) for the synthesis of complex 1. We evaluated lipid peroxidation, protein carbonylation, ROS generation, DNA damage and consequent apoptosis by complex 1 in exogenously added Cu(II) in human peripheral lymphocytes (simulate malignancy condition). Results showed that Cu(II)-complex 1 interaction leads to cell proliferation inhibition, apoptosis, ROS generation and DNA damage in human lymphocytes, and these effects were abrogated by cuprous chelator neocuproine and ROS scavengers (thiourea, catalase, SOD). This indicates that complex 1 cytotoxicity is due to redox cycling of copper to generate ROS which leads to pro-oxidant cell death in cancer cells. To further confirm our hypothesis, using the rat model of diethylnitrosamine (DEN) induced hepatocellular carcinoma; we showed that complex 1 mediates DNA breakage and cell death in isolated carcinoma cells. Membrane permeant copper chelator, neocuproine, and ROS scavengers inhibited the complex 1-mediated cellular DNA degradation and apoptosis. In summary, complex 1 anticancer activity is due to its copper chelation capability. These results will provide copper chelation as an effective targeted cancer treatment strategy for selective cytotoxic action against malignant cells without affecting normal cells.

Keywords: cancer treatment, copper chelation, ROS generation, DNA damage, redox cycling, apoptosis

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Authors: Fahimeh Palizban, Farshad Noravesh, Amir Hossein Saeidian, Mahya Mehrmohamadi

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In the recent decade, the emergence of liquid biopsy has significantly improved cancer monitoring and detection. Dying cells, including those originating from tumors, shed their DNA into the blood and contribute to a pool of circulating fragments called cell-free DNA. Accordingly, identifying the tissue origin of these DNA fragments from the plasma can result in more accurate and fast disease diagnosis and precise treatment protocols. Open chromatin regions are important epigenetic features of DNA that reflect cell types of origin. Profiling these features by DNase-seq, ATAC-seq, and histone ChIP-seq provides insights into tissue-specific and disease-specific regulatory mechanisms. There have been several studies in the area of cancer liquid biopsy that integrate distinct genomic and epigenomic features for early cancer detection along with tissue of origin detection. However, multimodal analysis requires several types of experiments to cover the genomic and epigenomic aspects of a single sample, which will lead to a huge amount of cost and time. To overcome these limitations, the idea of predicting OCRs from WGS is of particular importance. In this regard, we proposed a computational approach to target the prediction of open chromatin regions as an important epigenetic feature from cell-free DNA whole genome sequence data. To fulfill this objective, local sequencing depth will be fed to our proposed algorithm and the prediction of the most probable open chromatin regions from whole genome sequencing data can be carried out. Our method integrates the signal processing method with sequencing depth data and includes count normalization, Discrete Fourie Transform conversion, graph construction, graph cut optimization by linear programming, and clustering. To validate the proposed method, we compared the output of the clustering (open chromatin region+, open chromatin region-) with previously validated open chromatin regions related to human blood samples of the ATAC-DB database. The percentage of overlap between predicted open chromatin regions and the experimentally validated regions obtained by ATAC-seq in ATAC-DB is greater than 67%, which indicates meaningful prediction. As it is evident, OCRs are mostly located in the transcription start sites (TSS) of the genes. In this regard, we compared the concordance between the predicted OCRs and the human genes TSS regions obtained from refTSS and it showed proper accordance around 52.04% and ~78% with all and the housekeeping genes, respectively. Accurately detecting open chromatin regions from plasma cell-free DNA-seq data is a very challenging computational problem due to the existence of several confounding factors, such as technical and biological variations. Although this approach is in its infancy, there has already been an attempt to apply it, which leads to a tool named OCRDetector with some restrictions like the need for highly depth cfDNA WGS data, prior information about OCRs distribution, and considering multiple features. However, we implemented a graph signal clustering based on a single depth feature in an unsupervised learning manner that resulted in faster performance and decent accuracy. Overall, we tried to investigate the epigenomic pattern of a cell-free DNA sample from a new computational perspective that can be used along with other tools to investigate genetic and epigenetic aspects of a single whole genome sequencing data for efficient liquid biopsy-related analysis.

Keywords: open chromatin regions, cancer, cell-free DNA, epigenomics, graph signal processing, correlation clustering

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Authors: Sang-Myung Jung, Jeong Hyun Ju, Gwang Heum Yoon, Hwa Sung Shin

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Central nervous system (CNS) consists of neuronal cell and supporting cells. Astrocytes are the most common supporting cells and play roles in metabolism between neurons and blood vessel. For this function, engineered astrocytes have been studied as a therapeutic source for CNS injury. In neural tissue engineering, nanofiber has been suggested as an effective scaffold for providing structure and mechanical properties influencing physiology. Cellulose acetate (CA) has been investigated for material to fabricate scaffold because of its biocompatibility, biodegradability and fine thermal stability. In this research, CA nanofiber was modified via heat treatment and its effect on astrocyte activity was evaluated. Adhesion and viability of astrocyte were increased in proportion to stiffness. Additionally, expression of GFAP, a marker of astrocyte activation, was increased via stiffness of scaffold. This research suggests a simple modification method to change stiffness of CA nanofiber and shows cellular behavior affecting stiffness of three-dimensional scaffold independently. For the results, we highlight that the stiffness is a factor to regulate astrocyte activity.

Keywords: astrocyte, cellulose acetate, cell therapy, stiffness of scaffold

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Authors: A. Alvaredo , R. Guzman De Villoria, P. Castell, Juan P. Fernandez-Blazquez

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Since graphene was discovered in 2004, has been considered as superb material, due to its outstanding mechanical, electrical and thermal properties. Graphene has been incorporated as reinforcement in several high performance polymers in order to obtain a good balance of properties and to get new properties as thermal or electric conductivity. As well known, the properties of semicrystalline polymer and its composites depends heavily on degree of crystallinity. In this context, our research group has studied the crystallization behavior from amorphous state of PEEK/GNP composites. The monitoring of cold crystallization processes studied by time-resolved simultaneous wide-angle X-ray scattering (WAXS) and small-angle X-ray scattering (SAXS). These techniques allowed to get an extremely relevant information about the evolution of the morphology of the PEEK/GNP composites. In addition, the thermal evolution of cold crystallization was followed by differential scanning calorimetry (DSC) as well. The experimental results showed changes in crystallization kinetics and c parameter unit cell when adding graphene. The main aim of this work is to produce PEEK/GNP composites and characterize their morphology, unit cell parameters and crystallization kinetic.

Keywords: PEEK, graphene, synchrotron, cold crystallization

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Authors: Adel M. Mahmoud

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This Investigation was conducted to determine the productivity and water use efficiency for new sunflower genotypes. Ten sunflower genotypes were evaluated under drip irrigation using two treatments of. Results indicate that decreasing the amount of irrigation water from 1500 to 1130 mm/hectar significantly reduced all studied traits. Mutation (M1-63) surpassed all the other one genotypes in seed yield and WUE. Lines which gave the highest yield of the seed have water use efficiency under drought conditions higher than water use efficiency under normal irrigation. The lowest depression in seed yield due to drought conditions has been registered for Line 20, Line M1-63 and Sakha 53 genotypes (11 , 18 and 16 %, respectively). Genotypes (Line 20 , Line M1-63 and Sakha 53) are more tolerant to drought than others and we can used its in breeding program to develop sunflower hybrids suitable for cultivation under drought condition.

Keywords: sunflower genotypes, water use efficiency, mutation, inbred lines

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Authors: M. Yadegari, M. Nourbakhsh, N. Arbabzadeh

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The skeletal system injuries are of major importance. In addition, it is recommended to use materials for hard tissue repair in open or closed fractures. It is important to use complex minerals with a beneficial effect on hard tissue repair, stimulating cell growth in the bone. Materials that could help avoid bone fracture inflammatory reaction and speed up bone fracture repair are of utmost importance in the treatment of bone fractures. Similar to minerals, the inner eggshell membrane consists of carbohydrates, lipids, proteins with the high pH, high calcium absorptive capacity and with faster bone fracture repair ability. In the present radiographic survey, eggshell-derived bone graft substitutes were used for bone defect repair in 8 dog tibia, measuring bone density on the day of implant placement and 30 and 60 days after placement. In fact, the result of this study shows the difference in bone growth and misshapen bones between treatment and control sites. Cell growth was adequate in treatment sites and misshapen bones were less frequent here than in control sites.

Keywords: bone repair, eggshell powder, implant, radiography

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Authors: Edgar Ochoa, G. Torres-Villasenor

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Superplastic deformation is shown by materials with a fine grain size, usually less than 10 μm, when they are deformed within the strain rate range 10-5 10-1 s-1 at temperatures greater than 0.5Tm, where Tm is the melting point in Kelvin. According to the constitutive equation for superplastic flow, refinement of the grain size would be expected to increase the optimum strain rate and decrease the temperature required for superplastic flow. Ribbons of eutectic Ag-Cu and Bi-Cd alloys were manufactured by using a single roller melt-spinning technique to obtain a fine grain structure for later test in superplastic regime. The eutectics ribbons were examined by scanning electron microscopy and X-Ray diffraction, and the grain size was determined using the image analysis software ImageJ. The average grain size was less than 1 μm. Tensile tests were carried out from 10-4 to 10-1 s-1, at room temperature, to evaluate the superplastic behavior. The largest deformation was shown by the Bi-Cd eutectic ribbons, Ɛ=140 %, despite that these ribbons have a hexagonal unit cell. On the other hand, Ag-Cu eutectic ribbons have a minor grain size and cube unit cell, however they showed a lower deformation in tensile test under the same conditions than Bi-Cd ribbons. This is because the Ag-Cu grew in a strong cube-cube orientation relationship.

Keywords: eutectic ribbon, fine grain, superplastic deformation, cube-cube orientation

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Authors: Yi Sun, George Ed Rainger, Steve P. Watson

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Background: Beyond the classical roles in haemostasis and thrombosis, platelets are important in the initiation and development of various thrombo-inflammatory diseases. In atherosclerosis and deep vein thrombosis, for example, platelets bridge monocytes with endothelium and form heterotypic aggregates with monocytes in the circulation. This can alter monocyte phenotype by inducing their activation, stimulating adhesion and migration. These interactions involve cell surface receptor-ligand pairs on both cells. This list is likely incomplete as new interactions of importance to platelet biology are continuing to be discovered as illustrated by our discovery of PEAR-1 binding to FcεR1α. Results: We have developed a highly sensitive avidity-based assay to identify novel extracellular interactions among 126 recombinantly-expressed platelet cell surface and secreted proteins involved in platelet aggregation. In this study, we will use this method to identify novel platelet-monocyte interactions. We aim to identify ligands for orphan receptors and novel partners of well-known proteins. Identified interactions will be studied in preliminary functional assays to demonstrate relevance to the inflammatory processes supporting atherogenesis. Conclusions: Platelet-monocyte interactions are essential for the development of thromboinflammatory disease. Up until relatively recently, technologies only allow us to limit our studies on each individual protein interaction at a single time. These studies propose for the first time to study the cell surface platelet-monocyte interactions in a systematic large-scale approach using a reliable screening method we have developed. If successful, this will likely to identify previously unknown ligands for important receptors that will be investigated in details and also provide a list of novel interactions for the field. This should stimulate studies on developing alternative therapeutic strategies to treat vascular inflammatory disorders such as atherosclerosis, DVT and sepsis and other clinically important inflammatory conditions.

Keywords: membrane proteins, large-scale screening, platelets, recombinant expression

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Authors: C. Angel, S. Asha

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Despite the advances made in various new technologies, application of these technologies for agriculture still remains a formidable task, as it involves integration of diverse domains for monitoring the different process involved in agricultural management. Advances in ambient intelligence technology represents one of the most powerful technology for increasing the yield of agricultural crops and to mitigate the impact of water scarcity, climatic change and methods for managing pests, weeds, and diseases. This paper proposes a GPS-assisted, machine to machine solutions that combine information collected by multiple sensors for the automated management of paddy crops. To maintain the economic viability of paddy cultivation, the various techniques used in agriculture are discussed and a novel system which uses ambient intelligence technique is proposed in this paper. The ambient intelligence based agricultural system gives a great scope.

Keywords: ambient intelligence, agricultural technology, smart agriculture, precise farming

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Authors: M. Caramihai, Irina Severin

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This paper deals with the design of an intelligent control structure for a bioprocess of Hansenula polymorpha yeast cultivation. The objective of the process control is to produce biomass in a desired physiological state. The work demonstrates that the designed Hybrid Control Techniques (HCT) are able to recognize specific evolution bioprocess trajectories using neural networks trained specifically for this purpose, in order to estimate the model parameters and to adjust the overall bioprocess evolution through an expert system and a fuzzy structure. The design of the control algorithm as well as its tuning through realistic simulations is presented. Taking into consideration the synergism of different paradigms like fuzzy logic, neural network, and symbolic artificial intelligence (AI), in this paper we present a real and fulfilled intelligent control architecture with application in bioprocess control.

Keywords: bioprocess, intelligent control, neural nets, fuzzy structure, hybrid techniques

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Authors: Ali Bayram, Remzi Yiğiter

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Objective: Alzheimer's disease (AD), the most common cause of dementia, is a progressive neurodegenerative disease. At present, diagnosis of AD is rather late in the disease. Therefore, we attempted to find peripheral biomarkers for the early diagnosis of AD. Herein, we conducted a study to investigate the unc-51 like autophagy activating kinase-1 (ULK1) mRNA expression levels in human peripheral blood mononuclear cells from patients with Alzheimer's disease. Method: To determine whether ULK1 gene expression are altered in AD patients, we measured their gene expression in human peripheral blood cell in 50 patients with AD and 50 age and gender matched healthy controls by quantitative real-time PCR technique. Results: We found that both ULK1 gene expression in peripheral blood cell were significantly decreased in patients with AD as compared with controls (p <0.05). Lower levels of ULK1 gene expression were significantly associated with the increased risk for AD. Conclusions: Serine/threonine-protein kinase involved in autophagy in response to starvation. Acts upstream of phosphatidylinositol 3-kinase PIK3C3 to regulate the formation of autophagophores, the precursors of autophagosomes. Part of regulatory feedback loops in autophagy: acts both as a downstream effector and negative regulator of mammalian target of rapamycin complex 1 (mTORC1) via interaction with RPTOR. Activated via phosphorylation by AMPK and also acts as a regulator of AMPK by mediating phosphorylation of AMPK subunits PRKAA1, PRKAB2, and PRKAG1, leading to negatively regulate AMPK activity. May phosphorylate ATG13/KIAA0652 and RPTOR; however such data need additional evidences. Plays a role early in neuronal differentiation and is required for granule cell axon formation. Alzheimer is the most common neurodegenerative disease. Our results provide useful information that the ULK1 gene expression is decreased in the neurodegeneration and AD patients with, indicating their possible systemic involvement in AD.

Keywords: Alzheimer’s sisease, ULK1, mRNA expression, RT-PCR

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Authors: Hyunjung Lim, Jeonghun Nam, Hyuk Choi

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High-throughput separation is an essential technique for cancer research and diagnosis. Here, we propose a viscoelastic microfluidic device for sheathless, high-throughput isolation of circulating tumor cells (CTCs) from white blood cells. Here, we demonstrate a viscoelastic method for separation and concentration of CTCs using closed-loop microfluidics. Our device is a rectangular straight channel with a low aspect ratio. Also, to achieve high-efficiency, high-throughput processing, we used a polymer solution with low viscosity. At the inlet, CTCs and white blood cells (WBCs) were randomly injected into the microchannel. Due to the viscoelasticity-induced lateral migration to the equilibrium positions, large CTCs could be collected from the side outlet while small WBCs were removed at the center outlet. By recirculating the collected CTCs from the side outlet back to the sample reservoir, continuous separation and concentration of CTCs could be achieved with high separation efficiency (~ 99%). We believe that our device has the potential to be applied in resource-limited clinical settings.

Keywords: circulating tumor cell, closed-loop microfluidics, concentration, separation, viscoelastic fluid

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Authors: Dimitriya A. Mihaylova, Zlatka V. Valkova-Jarvis, Georgi L. Iliev

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One possible approach for maintaining the security of communication systems relies on Physical Layer Security mechanisms. However, in wireless time division duplex systems, where uplink and downlink channels are reciprocal, the channel estimate procedure is exposed to attacks known as pilot contamination, with the aim of having an enhanced data signal sent to the malicious user. The Shifted 2-N-PSK method involves two random legitimate pilots in the training phase, each of which belongs to a constellation, shifted from the original N-PSK symbols by certain degrees. In this paper, legitimate pilots’ offset values and their influence on the detection capabilities of the Shifted 2-N-PSK method are investigated. As the implementation of the technique depends on the relation between the shift angles rather than their specific values, the optimal interconnection between the two legitimate constellations is investigated. The results show that no regularity exists in the relation between the pilot contamination attacks (PCA) detection probability and the choice of offset values. Therefore, an adversary who aims to obtain the exact offset values can only employ a brute-force attack but the large number of possible combinations for the shifted constellations makes such a type of attack difficult to successfully mount. For this reason, the number of optimal shift value pairs is also studied for both 100% and 98% probabilities of detecting pilot contamination attacks. Although the Shifted 2-N-PSK method has been broadly studied in different signal-to-noise ratio scenarios, in multi-cell systems the interference from the signals in other cells should be also taken into account. Therefore, the inter-cell interference impact on the performance of the method is investigated by means of a large number of simulations. The results show that the detection probability of the Shifted 2-N-PSK decreases inversely to the signal-to-interference-plus-noise ratio.

Keywords: channel estimation, inter-cell interference, pilot contamination attacks, wireless communications

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Authors: K. J. Keerthi, Vasundhara Kamineni, A. Ravi Shanker, T. Rammurthy, A. Vijaya Lakshmi, Q. Hasan

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Pancreatic β-cells are the predominant insulin-producing cell types within the Islets of Langerhans and insulin is the primary hormone which regulates carbohydrate and fat metabolism. Apoptosis of β-cells or insufficient insulin production leads to Diabetes Mellitus (DM). Current therapy for diabetes includes either medical management or insulin replacement and regular monitoring. Replacement of β- cells is an attractive treatment option for both Type-1 and Type-2 DM in view of the recent paper which indicates that β-cells apoptosis is the common underlying cause for both the Types of DM. With the development of Edmonton protocol, pancreatic β-cells allo-transplantation became possible, but this is still not considered as standard of care due to subsequent requirement of lifelong immunosuppression and the scarcity of suitable healthy organs to retrieve pancreatic β-cell. Fetal pancreatic cells from abortuses were developed as a possible therapeutic option for Diabetes, however, this posed several ethical issues. Hence, in the present study Mesenchymal stem cells (MSCs) were differentiated into insulin producing cells which were isolated from Human Umbilical cord (HUC) tissue. MSCs have already made their mark in the growing field of regenerative medicine, and their therapeutic worth has already been validated for a number of conditions. HUC samples were collected with prior informed consent as approved by the Institutional ethical committee. HUC (n=26) were processed using a combination of both mechanical and enzymatic (collagenase-II, 100 U/ml, Gibco ) methods to obtain MSCs which were cultured in-vitro in L-DMEM (Low glucose Dulbecco's Modified Eagle's Medium, Sigma, 4.5 mM glucose/L), 10% FBS in 5% CO2 incubator at 37°C. After reaching 80-90% confluency, MSCs were characterized with Flowcytometry and Immunocytochemistry for specific cell surface antigens. Cells expressed CD90+, CD73+, CD105+, CD34-, CD45-, HLA-DR-/Low and Vimentin+. These cells were differentiated to β-cells by using H-DMEM (High glucose Dulbecco's Modified Eagle's Medium,25 mM glucose/L, Gibco), β-Mercaptoethanol (0.1mM, Hi-Media), basic Fibroblast growth factor (10 µg /L,Gibco), and Nicotinamide (10 mmol/L, Hi-Media). Pancreatic β-cells were confirmed by positive Dithizone staining and were found to be functionally active as they released 8 IU/ml insulin on glucose stimulation. Isolating MSCs from usually discarded, abundantly available HUC tissue, expanding and differentiating to β-cells may be the most feasible cell therapy option for the millions of people suffering from DM globally.

Keywords: diabetes mellitus, human umbilical cord, mesenchymal stem cells, differentiation

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Authors: Mohanad Alhabo, Li Zhang, Naveed Nawaz

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In this paper, a non-cooperative game method is formulated where all players compete to transmit at higher power. Every base station represents a player in the game. The game is solved by obtaining the Nash equilibrium (NE) where the game converges to optimality. The proposed method, named Power Efficient Handover Game Theoretic (PEHO-GT) approach, aims to control the handover in dense small cell networks. Players optimize their payoff by adjusting the transmission power to improve the performance in terms of throughput, handover, power consumption and load balancing. To select the desired transmission power for a player, the payoff function considers the gain of increasing the transmission power. Then, the cell selection takes place by deploying Technique for Order Preference by Similarity to an Ideal Solution (TOPSIS). A game theoretical method is implemented for heterogeneous networks to validate the improvement obtained. Results reveal that the proposed method gives a throughput improvement while reducing the power consumption and minimizing the frequent handover.

Keywords: energy efficiency, game theory, handover, HetNets, small cells

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Authors: Ahmed M. Hjazi, Bader M. Hjazi

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Protein 4.1 is a crucial component of complex interactions between the cytoskeleton and other junctional complex proteins. When the gene encoding this protein is altered, resulting in reduced expression, or when the protein is absent, the red cell undergoes a significant structural change. This research aims to achieve a deeper comprehension of the biochemical effects of red cell protein deficiency. A Tandem Mass Spectrometry Analysis (TMT-MS/MS) of patient cells lacking protein 4.1 compared to three healthy controls was achieved by the Proteomics Institute of the University of Bristol. The SDS-PAGE and Western blotting were utilized on the original patient sample and controls to partially confirm TMT MS/MS data analysis of the protein-4.1-deficient cells. Compared to healthy controls, protein levels in samples lacking protein 4.1 had a significantly higher concentration of proteins that probably originated from reticulocytes. This could occur if the patient has an elevated reticulocyte count. The increase in chaperone and reticulocyte-associated proteins was most notable in this study. This may result from elevated quantities of reticulocytes in patients with hereditary elliptocytosis.

Keywords: hereditary elliptocytosis, protein 4.1, red cells, tandem mass spectrometry data.

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Authors: Yung-Chung Chuang

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Current situation and accumulated development of archaeological sites have very high impacts on the preservation value of the site. This research set 3 archaeological sites in Taiwan as study areas. Assessment of the degree of destruction of cultural layers due to land use change and geomorphological change were conducted with aerial photographs (1976-1978; 2016-2017) and digital aerial survey technology on 2D and 3D geographic information system platforms. The results showed that the archaeological sites were all seriously influenced due to the high land use intensity between 1976-2017. Geomorphological changes caused by human cultivation and engineering construction were main causes of site destruction, especially in private lands. Therefore, urban planning methods for land acquisition or land regulation are necessary.

Keywords: archaeological sites, accumulated development, destruction of cultural layers, geomorphological changes

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Authors: Sam Weber, Deepak Mishra, Susan Wilde, Elizabeth Kramer

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The frequency and severity of cyanobacteria harmful blooms (CyanoHABs) have been increasing over time, with point and non-point source eutrophication and shifting climate paradigms being blamed as the primary culprits. Excessive nutrients, warm temperatures, quiescent water, and heavy and less regular rainfall create more conducive environments for CyanoHABs. CyanoHABs have the potential to produce a spectrum of toxins that cause gastrointestinal stress, organ failure, and even death in humans and animals. To promote enhanced, proactive CyanoHAB management, risk modeling using geospatial tools can act as predictive mechanisms to supplement current CyanoHAB monitoring, management and mitigation efforts. The risk maps would empower water managers to focus their efforts on high risk water bodies in an attempt to prevent CyanoHABs before they occur, and/or more diligently observe those waterbodies. For this research, exploratory spatial data analysis techniques were used to identify the strongest predicators for CyanoHAB blooms based on remote sensing-derived cyanobacteria cell density values for 771 waterbodies in the Georgia Piedmont and landscape characteristics of their watersheds. In-situ datasets for cyanobacteria cell density, nutrients, temperature, and rainfall patterns are not widely available, so free gridded geospatial datasets were used as proxy variables for assessing CyanoHAB risk. For example, the percent of a watershed that is agriculture was used as a proxy for nutrient loading, and the summer precipitation within a watershed was used as a proxy for water quiescence. Cyanobacteria cell density values were calculated using atmospherically corrected images from the European Space Agency’s Sentinel-2A satellite and multispectral instrument sensor at a 10-meter ground resolution. Seventeen explanatory variables were calculated for each watershed utilizing the multi-petabyte geospatial catalogs available within the Google Earth Engine cloud computing interface. The seventeen variables were then used in a multiple linear regression model, and the strongest predictors of cyanobacteria cell density were selected for the final regression model. The seventeen explanatory variables included land cover composition, winter and summer temperature and precipitation data, topographic derivatives, vegetation index anomalies, and soil characteristics. Watershed maximum summer temperature, percent agriculture, percent forest, percent impervious, and waterbody area emerged as the strongest predictors of cyanobacteria cell density with an adjusted R-squared value of 0.31 and a p-value ~ 0. The final regression equation was used to make a normalized cyanobacteria cell density index, and a Jenks Natural Break classification was used to assign waterbodies designations of low, medium, or high risk. Of the 771 waterbodies, 24.38% were low risk, 37.35% were medium risk, and 38.26% were high risk. This study showed that there are significant relationships between free geospatial datasets representing summer maximum temperatures, nutrient loading associated with land use and land cover, and the area of a waterbody with cyanobacteria cell density. This data analytics approach to CyanoHAB risk assessment corroborated the literature-established environmental triggers for CyanoHABs, and presents a novel approach for CyanoHAB risk mapping in waterbodies across the greater southeastern United States.

Keywords: cyanobacteria, land use/land cover, remote sensing, risk mapping

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Authors: Mauricio Quintero Angel, Andrés A. Duque Nivia, Carlos H. Fajardo Toro

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This paper analyzes two approaches of sustainability, the weak and strong, considering a case of study of oil palm production for an industry of biodegradable detergent. In this case, a company demand the oil palm as the active element for washing and through its trademark aims to supply 10% of the Colombian market of washing powders. Under each approach the economic and ecological implications of the palm oil production and especially the implications for crop management are described. The crop production under the weak sustainability implies plantations, intensive use of agrochemicals and the inclusion of new areas of cultivation as the market grows. Under the strong sustainability the production system is limited by the productive vocation of the ecosystem, so that new approaches and creativity for making viable the nature conservancy and the business development are require.

Keywords: agriculture, environmental impacts, oil palm, strong sustainability, weak sustainability

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Authors: Humayun Kabir, Amirul Hasan, Yu Miyaoka, Makiko Yamaguchi, Chisaki Kadota, Kazuaki Takehara

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From field isolates of Newcastle disease virus (NDV) in Japan, one avirulent strain, APMV/northern pintail/Japan/Aomori/2003 (dk-Aomori/03, NDV 261), was selected for its excellent thermostability, and the strain was heat-treated at 56℃ temperatures for 30 min with each passage into Vero cells to maintain thermostability and to adapt Vero cells. After serial 20 passages in Vero cells, it was named NDV Vero20. When growth curves were tested in Vero cells, NDV Vero20 grew well to compare the original NDV261. The HN gene was sequenced, and found motifs that show thermostability. The intracerebral pathogenicity index (ICPI) test score was 0. The thermostability of the virus was confirmed by storing it at different temperatures, including at 37°C. When susceptible chicks were inoculated with NDV Vero20 through eye drops, induced adequate levels of antibody were measured using a serum neutralization test. The results showed that NDV Vero20, a vaccine candidate strain is thermostable, Vero cell adapted, and has immunogenic potential, which would make as an alternative to the traditional embryonated chicken eggs-based vaccine.

Keywords: Newcastle disease virus, thermostability, vaccine, Vero cell adaptability

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Authors: Caleb B. Sagwa, Tobias O. Okeno, Alexander K. Kahi

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Dairy cattle production plays an important role in the Kenyan economy. However, high incidences of mastitis is a major setback to the productivity in this industry. The current dairy cattle breeding objective in Kenya does not include mastitis resistance, mainly because the economic value of mastitis resistance has not been determined. Therefore this study aimed at estimating the economic value of mastitis resistance in dairy cattle in Kenya. Initial input parameters were obtained from literature on dairy cattle production systems in the tropics. Selection index methodology was used to derive the economic value of mastitis resistance. Somatic cell count (SCC) was used an indicator trait for mastitis resistance. The economic value was estimated relative to milk yield (MY). Economic values were assigned to SCC in a selection index such that the overall gain in the breeding goal trait was maximized. The option of estimating the economic value for SCC by equating the response in the trait of interest to its index response was considered. The economic value of mastitis resistance was US $23.64 while maximum response to selection for MY was US $66.01. The findings of this study provide vital information that is a pre-requisite for the inclusion of mastitis resistance in the current dairy cattle breeding goal in Kenya.

Keywords: somatic cell count, milk quality, payment system, breeding goal

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Authors: Ashagrie Getnet Flattie

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Line-of-sight (LOS) information, data rates, good quality, and flexible network service are limited by the fact that, for the duration of any given connection, they experience severe variation in signal strength due to fading and path loss. Wireless system faces major challenges in achieving wide coverage and capacity without affecting the system performance and to access data everywhere, all the time. In this paper, the cell coverage and edge rate of different Multiple-input multiple-output (MIMO) schemes in 20 MHz Long Term Evolution (LTE) system under Unmanned Air Vehicles (UAV) platform are investigated. After some background on the enormous potential of UAV, MIMO, and LTE in wireless links, the paper highlights the presented system model which attempts to realize the various benefits of MIMO being incorporated into UAV platform. The performances of the three MIMO LTE schemes are compared with the performance of 4x4 MIMO LTE in UAV scheme carried out to evaluate the improvement in cell radius, BER, and data throughput of the system in different morphology. The results show that significant performance gains such as bit error rate (BER), data rate, and coverage can be achieved by using the presented scenario.

Keywords: LTE, MIMO, path loss, UAV

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Authors: F. L. Benu, I. W. Mudita, R. L. Natonis

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East Nusa Tenggara, together with part of East Java, West Nusa Tenggara, and Maluku, has been included as part of global drylands defined according to the ratio of annual precipitation (P) and annual potential evaporation (PET) and major vegetation types of grassland and savannah ecosystems. These tropical drylands are unique because, whereas drylands in other countries are mostly continental, here they are archipelagic. These archipelagic drylands are also unique in terms of being included because of more on their major vegetation types than of their P/PET ratio. Slash-and-burn cultivation and free roaming animal husbandry are two major livelihoods being widely practiced, along with alternative seasonal livelihood such as traditional fishing. Such livelihoods are vulnerable in various respects, especially because of drought, which becomes more unpredictable in the face of climate changes. To cope with such vulnerability, semi-intensive farming using drip irrigation is implemented as an appropriate technology with the goal of promoting a more sustainable alternative to the existing livelihoods. The implementation was started in 2016 with a pilot system at the university field laboratory in Kupang in which various designs of installation were tested. The modified system consisting of an uplifted water reservoir and solar-powered pump was tested in Papela, the District of Rote-Ndao, in 2017 to convince fishermen who had been involved in illegal fishing in Australia-Indonesia transboundary waters, to adopt small-scale farming as a more sustainable alternative to their existing livelihoods. The system was again tested in a larger coverage in Oesena, the District of Kupang, in 2018 to convince slash-and-burn cultivators to adopt an environmentally friendlier cultivation system. From the implementation of the modified system in both sites, the participating fishermen in Papela were able to manage the system under tight water supply to grow chili pepper, tomatoes, and watermelon and the slash-and-burn cultivators in Oesena to grow chili pepper in a more efficient water use than water use in a conventional irrigation system. The gross margin obtained from growing chili pepper, tomatoes, and watermelon in Papela and from growing chili pepper in Oesena showed that small-scale farming using drip irrigation system was a promising alternative to local people in generating cash income to support their livelihoods. However, before promoting this appropriate technology as a more sustainable alternative to the existing livelihoods elsewhere in the region, better understanding on social-related contexts of the implementation is needed.

Keywords: archipelagic drylands, drip irrigation system, East Nusa Tenggara, sustainable livelihoods

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Authors: Md. Maksudur Rahman Khan, Chee Wai Woon, Huei Ruey Ong, Vignes Rasiah, Chin Kui Cheng, Kar Min Chan, E. Baranitharan

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The present work represents a preparation of non-precious iron-based electrocatalyst (FeN) for ORR in air-cathode microbial fuel cell by pyrolysis treatment. Iron oxalate which recovered from the industrial wastewater and Phenanthroline (Phen) were used as the iron and nitrogen precursors, respectively in preparing FeN catalyst. The performance of as prepared catalyst (FeN) was investigated in a single chambered air cathode MFC in which anaerobic sludge was used as inoculum and palm oil mill effluent as substrate. The maximum open circuit potential (OCV) and the highest power density recorded were 0.543 V and 4.9 mW/m2, respectively. Physical characterization of FeN was elucidated by using Brunauner Emmett Teller (BET), X-Ray Diffraction (XRD) analysis and Field Emission Scanning Electron Microscopy (FESEM) while the electrochemical properties were characterized by cyclic voltammetry (CV) analysis. The presence of biofilm on anode surface was examined using FESEM and confirmed using Infrared Spectroscopy and Thermogravimetric Analysis. The findings of this study demonstrated that FeN is electrochemically active and further modification is needed to increase the ORR catalytic activity.

Keywords: iron based catalyst, microbial fuel cells, oxygen reduction reaction, palm oil mill effluent

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Authors: Nonita Sarin, K.J.Singh, Anuj Kumar, Davinder Singh

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Polymeric 3D hydrogels have pivotal role in bone tissue regeneration applications. Hydrogels behave similar to the living tissues because they have large water imbibing capacity in swollen state and adjust their shape according to the tissues during tissue formation after implantation. On the other hand, hydrogels are very soft, fragile and lack mechanical strength. Incorporation of bioceramics can improve mechanical strength. Furthermore, bioceramics synthesized by sol gel technique may enhance the apatite formation and degradation rates which can lead to the increase in faster rates for new bone and tissue regeneration. Simulated body fluid (SBF) induces the poly-condensation of silanol groups which leads to formation of silica matrix and provide active sites for the precipitation of Ca2+ and PO43- ions to form apatite layer which is similar to mineral form of bone. Therefore, authors have synthesized bioceramic incorporated Polyacrylamide-carboxymethylcellulose hydrogels by free radical polymerization and bioceramic compositions of xSrO-(36-x)CaO-45SiO2-ySeO3-(12-y)P2O5-7MgO (where x=0,4 and y=0,2 mol%) were synthesized by sol gel technique. Bioceramics incorporated in polymer matrix induces quicker apatite formation during immersion in SBF by raising the pH with the release of alkaline ions during ion exchange process and the apatite formation takes place in alkaline medium. The behavior of samples PABC-0 (without bioceramics) and PABC-20 (with 20 wt% bioceramics) were evaluated by X-Ray Diffraction and FTIR. In term of bioactivity, it was observed that PABC-20 has shown hydroxyapatite (HA) formation on 1st day of immersion whereas, PABC-0 was shown apatite formation on 7th day of immersion in SBF. The rapid rate of HA growth on 1st day of immersion in SBF signifies easy regeneration of damaged bone tissues. Degradation studies have been undertaken in Phosphate Buffer Saline and PABC-20 exhibited slower degradation rate up to 9%as compared to PABC-0 up to 18%. Slower degradation rate is suitable for new tissue regeneration and cell attachment. Also, Zeta potential studies have been employed to check the surface charge and it has been observed that samples carry negative charge when immersed in SBF. In addition, the swelling test of the samples have been performed and relative swelling ratio % observed for PABC-0 is 607% and PABC-20 is 305%. This indicates that the incorporation of bioceramics leads to the filling up of the voids in between the polymer matrix which in result reduces porosity and increase the mechanical strength by filling the voids. The porosity of PABC-0 is 84% and PABC-20 is 72%. PABC-20 sample demonstrates that bioceramics incorporation reduce the porosity and improves mechanical strength. Also, maximum in vitro cell viability up to 98% with MG63 cell line has been observed which indicate that the bioceramic incorporated hydrogel(PABC-20) provide the alkaline medium which is suitable environment for cell growth.

Keywords: hydrogels, hydroxyapatite, MG63 cell line, zeta potential

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Authors: G. Eom, H. Kim, A. Hwang, T. Kang, B. Kim

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Telomerase activity is overexpressed in over 85% of human cancers while suppressed in normal somatic cells. Telomerase has been attracted as a universal cancer biomarker. Therefore, the development of effective telomerase activity detection methods is urgently demanded in cancer diagnosis and therapy. Herein, we report a nanogap-rich Au nanowire (NW) surface-enhanced Raman scattering (SERS) sensor for detection of human telomerase activity. The nanogap-rich Au NW SERS sensors were prepared simply by uniformly depositing nanoparticles (NPs) on single-crystalline Au NWs. We measured SERS spectra of methylene blue (MB) from 60 different nanogap-rich Au NWs and obtained the relative standard deviation (RSD) of 4.80%, confirming the superb reproducibility of nanogap-rich Au NW SERS sensors. The nanogap-rich Au NW SERS sensors enable us to detect telomerase activity in 0.2 cancer cells/mL. Furthermore, telomerase activity is detectable in 7 different cancer cell lines whereas undetectable in normal cell lines, which suggest the potential applicability of nanogap-rich Au NW SERS sensor in cancer diagnosis. We expect that the present nanogap-rich Au NW SERS sensor can be useful in biomedical applications including a diverse biomarker sensing.

Keywords: cancer biomarker, nanowires, surface-enhanced Raman scattering, telomerase

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Authors: Ji-Young Choi, Jungjin Kim, Sang-Sun Yun, Sangmee Ahn Jo

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Intracellular adhesion molecule1 (ICAM1) is a mediator of inflammation and involved in adhesion and transmigration of leukocytes to endothelial cells, resulting in enhancement of brain inflammation. We hypothesized that increase of ICAM1 expression in endothelial cells is an early step in the pathogenesis of brain diseases such as Alzheimer’s disease. Here, we report that ICAM1 expression is regulated by pro-inflammatory cytokine TNFa in human microvascular endothelial cell (HBMVEC). TNFa significantly increased ICAM1 mRNA and protein levels at the concentrations showing no cell toxicity. This increase was also shown in micro vessels of mouse brain 24 hours after treatment with TNFa (8 mg/kg, i.v). We then investigated the epigenetic mechanism involved in the induction of ICAM1 expression. Chromatin immunoprecipitation assay revealed that TNFa reduced methylation of histone3K9 (H3K9-2me) and histone3K27 (H3K27-3me), well-known modification as gene suppression, with in the ICAM1 promoter region. However, acetylation of H3K9 and H3K14, well-known modification as gene activation, was not changed by TNFa. Treatment of BIX01294, a specific inhibitor of histone methyltransferase G9a responsible for H3K9-2me, dramatically increased in ICAM1 mRNA and protein levels and overexpression of G9a gene suppressed TNFa-induced ICAM1 expression. In contrast, GSK126, an inhibitor of histone methyltransferase EZH2 responsible for H3K27-3me and valproic acid, an inhibitor of histone deacetylase (HDAC) did not affect ICAM1 expression. These results suggested that histone3 methylation is involved in ICAM1 repression. Moreover, TNFa or BIX01294-induced ICAM induction resulted in both enhancements in adhesion and transmigration of leukocyte on endothelial cell. This study demonstrates that TNFa upregulates ICAM1 expression through H3K9-2me and H3K27-3me within the ICAM1 promoter region, in which G9a is likely to play a pivotal role in ICAM1 transcription. Our study provides a novel mechanism for ICAM1 transcription regulation in HBMVEC.

Keywords: ICAM1, TNFa, HBMVEC, H3K9-2me

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