Search results for: exponential expression profile

Authors: P. Narrima, C. Y. Looi, M. A. Mohd, H. M. Ali

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Persea declinata (Bl.) Kosterm is a member of the Lauraceae family, widely distributed in Southeast Asia. It is from the same genus with avocado (Persea americana Mill), which is widely consumed as food and for medicinal purposes. In the present study, we examined the anticancer properties of Persea declinata (Bl.) Kosterm bark methanolic crude extract (PDM). PDM exhibited a potent antiproliferative effect in MCF-7 human breast cancer cells, with an IC50 value of 16.68 µg/mL after 48h of treatment. We observed that PDM caused cell cycle arrest and subsequent apoptosis in MCF-7 cells, as exhibited by increased population at G0/G1 phase, higher lactate dehydrogenase (LDH) release, and DNA fragmentation. Mechanistic studies showed that PDM caused significant elevation in ROS production, leading to perturbation of mitochondrial membrane potential, cell permeability, and activation of caspases-3/7. On the other hand, real-time PCR and Western blot analysis showed that PDM treatment increased the expression of the proapoptotic molecule, Bax, but decreased the expression of prosurvival proteins, Bcl-2 and Bcl-xL, in a dose-dependent manner. These findings imply that PDM could inhibit proliferation in MCF-7 cells via cell cycle arrest and apoptosis induction, indicating its potential as a therapeutic agent worthy of further development.

Keywords: antiproliferative, apoptosis, MCF-7 human breast cancer, Persea declinata

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Authors: J. P. Lavande, A. V.Chandewar

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Mouth dissolving tablet is the speedily growing and highly accepted drug delivery system. This study was aimed at development of Ketorolac Tromethamine mouth dissolving tablet (MDTs), which can disintegrate or dissolve rapidly once placed in the mouth. Conventional Ketorolac tromethamine tablet requires water to swallow it and has limitation like low disintegration rate, low solubility etc. Ketorolac Tromethamine mouth dissolving tablets (formulation) consist of super-disintegrate like Heat Modified Karaya Gum, Co-treated Heat Modified Agar & Filler microcrystalline cellulose (MCC). The tablets were evaluated for weight variation, friability, hardness, in vitro disintegration time, wetting time, in vitro drug release profile, content uniformity. The obtained results showed that low weight variation, good hardness, acceptable friability, fast wetting time. Tablets in all batches disintegrated within 15-50 sec. The formulation containing superdisintegrants namely heat modified karaya gum and heat modified agar showed better performance in disintegration and drug release profile.

Keywords: mouth dissolving tablet, Ketorolac tromethamine, disintegration time, heat modified karaya gum, co-treated heat modified agar

Procedia PDF Downloads 287

Authors: Chanchai Supaartagorn

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E-retailing is the sale of goods online that takes place over the Internet. The Internet has shrunk the entire World. The world e-retailing is growing at an exponential rate in the Americas, Europe, and Asia. However, e-retailing costs require expensive investment, such as hardware, software, and security systems. Cloud computing technology is internet-based computing for the management and delivery of applications and services. Cloud-based e-retailing application models allow enterprises to lower their costs with their effective implementation of e-retailing activities. In this paper, we describe the concept of cloud computing and present the architecture of cloud computing, combining the features of e-retailing. In addition, we propose a strategy for implementing cloud computing with e-retailing. Finally, we explain the benefits from the architecture.

Keywords: architecture, cloud computing, e-retailing, internet-based

Procedia PDF Downloads 399

Authors: Adriana-Gabriela Plaiasu, Catalin Marian Ducu

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The interest in the unique properties associated with materials having structures on a nanometer scale has been increasing at an exponential rate in last decade. Among the functional mineral compounds such as perovskite (CaTiO3), rutile (TiO2), CaF2, spinel (MgAl2O4), wurtzite (ZnS), zincite (ZnO) and the cupric oxide (CuO) has been used in numerous applications such as catalysis, semiconductors, batteries, gas sensors, biosensors, field transistors and medicine. The Solar Physical Vapor Deposition (SPVD) presented in the paper as elaboration method is an original process to prepare nanopowders working under concentrated sunlight in 2kW solar furnaces. The influence of the synthesis parameters on the chemical and microstructural characteristics of zinc and manganese oxides synthesized nanophases has been systematically studied using XRD, TEM and SEM.

Keywords: characterization, morphological, nano-oxides, structural

Procedia PDF Downloads 284

Authors: O. J. Osunkeye, P. O. Fakolade, B. E. Olorede

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Broilers productions depend on the provision of adequate and goo quality feed containing all the nutrients, including proteins, carbohydrate, fats, vitamins, minerals and water. All these nutrients have to be provided at a required amount to support maximum productivity and normal physiological functions and demands. Among these nutrients proteins are particularly important, since they are essential for meat and muscle production, optimum growth and health status. Poultry production industry in the developing countries is been threatened because of the over dependency on Soybean meal as one of the key/major conventional protein stuff for feeding livestock. Even the competition between man and livestock for Soybean and other protein sources made the price of this feed stuff to be on the increase. Hence the needs to seek for an alternative feed stuff which is cheap and less competitive. This study showed the blood profile, organ and carcass characteristics and performance of broilers fed with Cowpea Testa Meal (CTM) based diets. Four diets were formulated with Cowpea Testa replacing Soybean at 0%, 15%, 30%, and 50% graded levels. One hundred and twenty day-old unsexed broiler birds were allotted to these four treatments with 3 replicates of 10 birds per replicate. The results showed no significant differences in all the haematological parameters measured (P>0.05), the serum metabolites analysis revealed significant different in Cholesterol (99.8 mg/dl, 112.84 mg/dl, 131.07 mg/dl and 97.66 mg/dl respectively) (P<0.05) among others. There were significant differences within the diets for average daily weight gain, average feed intake and feed to gain ratio. The birds on control (0%) and CTM gained more weight than those fed with 30% and 50% CTM diets. The organs and carcass primal cuts of the broilers expressed significant different for the spleen (0.12 g, 0.09 g, 0.11 g and 0.14 g respectively), lungs (0.97 g, 0.72 g, 0.77 g and 1.01g respectively) and proventriculus (0.96 g, 0.99 g, 0.81 g and 0.85 g respectively) (P<0.05). For the carcass, there were no significant differences (P<0.05) in the breast, thigh, drumstick, wing and neck except for the Back (21.27 g, 21.04 g, 17.71 g, and 17.89 g respectively). In conclusion, CTM inclusion in broiler’s diet could be used as an alternative feed stuff in replacement of Soybean meal up to 15% without any adverse effects as revealed by the blood profile and to increase the growth performance of the birds.

Keywords: physiological functions, cholesterol, blood profiles, CTM and carcass analysis

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Authors: Surajit Bhattacharya, Daniel Veltri, Atit A. Patel, Daniel N. Cox

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miRNAs have emerged as key post-transcriptional regulators of gene expression, however identification of biologically-relevant target genes for this epigenetic regulatory mechanism remains a significant challenge. To address this knowledge gap, we have developed a novel tool in R, Intra-miR-ExploreR, that facilitates integrated discovery of miRNA targets by incorporating target databases and novel target prediction algorithms, using statistical methods including Pearson and Distance Correlation on microarray data, to arrive at high confidence intragenic miRNA target predictions. We have explored the efficacy of this tool using Drosophila melanogaster as a model organism for bioinformatics analyses and functional validation. A number of putative targets were obtained which were also validated using qRT-PCR analysis. Additional features of the tool include downloadable text files containing GO analysis from DAVID and Pubmed links of literature related to gene sets. Moreover, we are constructing interaction maps of intragenic miRNAs, using both micro array and RNA-seq data, focusing on neural tissues to uncover regulatory codes via which these molecules regulate gene expression to direct cellular development.

Keywords: miRNA, miRNA:mRNA target prediction, statistical methods, miRNA:mRNA interaction network

Procedia PDF Downloads 515

Authors: Odinah Cuartero-Enteria, Aive Pecasales, Jhadly Philip Buniel, Christian Joy Vega, Shiela Estubo

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In the Philippines, Mindanao Island has the cheapest electricity because of the hydroelectric plants. Due to the rapid increase of the electricity consumption which the sources of electricity cannot support, it causes rotating brownout during summer season. This study investigated the household energy usage and practices in the rural areas of northern part of the Mindanao Island, Philippines. The questionnaire that includes the respondents’ profile and their common practices in energy consumptions was used as a tool in gathering the data. Several households were subjected to the survey. Results show energy consumption is not dependent on the profile of the respondents. It was observed that most of the families prefer to use energy saving methods of reducing electricity consumption. The main energy saving methods are unplugging unused home appliances, using of compact fluorescent bulb and energy-efficient gadgets, and using high electricity consumption appliances by schedule. Based on the results, the households in the rural areas know the practices of reducing electricity consumption. However, it is highly recommended that concern agencies should initiate information dissemination and strict implementation of well-formulated energy conservation practices all over the areas in Mindanao.

Keywords: Philippines, Mindanao island, rural areas, households, energy usages, practices

Procedia PDF Downloads 384

Authors: Samantha Shallop, Hakinya Karra, Tytus Bernas, Gladys Shaw, Gretchen Neigh, Jeffrey Dupree, Mathula Thangarajh

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Abnormal neuronal homeostasis is considered a structural correlate of cognitive deficits in Duchenne Muscular Dystrophy. Neurons are highly polarized cells with multiple dendrites but a single axon. Trafficking of cellular organelles are highly regulated, with the cargo in the somatodendritic region of the neuron not permitted to enter the axonal compartment. We investigated the molecular mechanisms that regular organelle trafficking in neurons using a multimodal approach, including high-resolution structural illumination, proteomics, immunohistochemistry, and computational modeling. We investigated the expression of ankyrin-G, the master regulator controlling neuronal polarity. The expression of ankyrin G and the morphology of the axon initial segment was profoundly abnormal in the CA1 hippocampal neurons in the mdx52 animal model of DMD. Ankyrin-G colocalized with kinesin KIF5a, the anterograde protein transporter, with higher levels in older mdx52 mice than younger mdx52 mice. These results suggest that the functional trafficking from the somatodendritic compartment is abnormal. Our data suggests that dystrophin deficiency compromised neuronal homeostasis via ankyrin-G-based mechanisms.

Keywords: neurons, axonal transport, duchenne muscular dystrophy, organelle transport

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Authors: Lenon M. Pereira, Helen J. Khoury, Marcos E. A. Andrade, Dean L. Cutajar, Vinicius S. M. Barros, Anatoly B. Rozenfeld

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With the increase of beam widths and the advent of multiple-slice and helical scanners, concerns related to the current dose measurement protocols and instrumentation in computed tomography (CT) have arisen. The current methodology of dose evaluation, which is based on the measurement of the integral of a single slice dose profile using a 100 mm long cylinder ionization chamber (Ca,100 and CPPMA, 100), has been shown to be inadequate for wide beams as it does not collect enough of the scatter-tails to make an accurate measurement. In addition, a long ionization chamber does not offer a good representation of the dose profile when tube current modulation is used. An alternative approach has been suggested by translating smaller detectors through the beam plane and assessing the accumulated dose trough the integral of the dose profile, which can be done for any arbitrary length in phantoms or in the air. For this purpose, a MOSFET dosimeter of small dosimetric volume was used. One of its recently designed versions is known as the MOSkin, which is developed by the Centre for Medical Radiation Physics at the University of Wollongong, and measures the radiation dose at a water equivalent depth of 0.07 mm, allowing the evaluation of skin dose when placed at the surface, or internal point doses when placed within a phantom. Thus, the aim of this research was to characterize the response of the MOSkin dosimeter for X-ray CT beams and to evaluate its application for the accumulated dose assessment. Initially, tests using an industrial x-ray unit were carried out at the Laboratory of Ionization Radiation Metrology (LMRI) of Federal University of Pernambuco, in order to investigate the sensitivity, energy dependence, angular dependence, and reproducibility of the dose response for the device for the standard radiation qualities RQT 8, RQT 9 and RQT 10. Finally, the MOSkin was used for the accumulated dose evaluation of scans using a Philips Brilliance 6 CT unit, with comparisons made between the CPPMA,100 value assessed with a pencil ionization chamber (PTW Freiburg TW 30009). Both dosimeters were placed in the center of a PMMA head phantom (diameter of 16 cm) and exposed in the axial mode with collimation of 9 mm, 250 mAs and 120 kV. The results have shown that the MOSkin response was linear with doses in the CT range and reproducible (98.52%). The sensitivity for a single MOSkin in mV/cGy was as follows: 9.208, 7.691 and 6.723 for the RQT 8, RQT 9 and RQT 10 beams qualities respectively. The energy dependence varied up to a factor of ±1.19 among those energies and angular dependence was not greater than 7.78% within the angle range from 0 to 90 degrees. The accumulated dose and the CPMMA, 100 value were 3,97 and 3,79 cGy respectively, which were statistically equivalent within the 95% confidence level. The MOSkin was shown to be a good alternative for CT dose profile measurements and more than adequate to provide accumulated dose assessments for CT procedures.

Keywords: computed tomography dosimetry, MOSFET, MOSkin, semiconductor dosimetry

Procedia PDF Downloads 315

Authors: Junichi Hosoi

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Epidermal Langerhans cells reside in upper layer of epidermis and play a role in immune surveillance. The finding of the close association of nerve endings to Langerhans cells triggered the research on systemic regulation of Langerhans cells. They disappear from epidermis after exposure to environmental and internal stimuli and reappear about a week later. Myeloid progenitor cells are assumed to be one of the sources of Langerhans cells. We examined the effects of cortisol on the reappearance of Langerhans cells in vitro. Cord-blood derived CD34-positive cells were cultured in the medium supplemented with stem cell factor/Flt3 ligand/granulocyte macrophage-colony stimulating factor/tumor necrosis factor alpha/bone morphologic protein 7/transforming growth factor beta in the presence or absence of cortisol. Cells were analyzed by flow cytometry for CD1a (cluster differentiation 1a), a marker of Langerhans cells and dermal dendritic cells, and CD39 (cluster differentiation factor 39), extracellular adenosine triphosphatase. Both CD1a-positive cells and CD39-positive cells were decreased by treatment with cortisol (suppression by 35% and 22% compared to no stress hormone, respectively). Differentiated Langerhans cells are attracted to epidermis by chemokines that are secreted from keratinocytes. Epidermal keratinocytes were cultured in the presence or absence of cortisol and analyzed for the expression of CCL2 (C-C motif chemokine ligand 2) and CCL20 (C-C motif chemokine ligand 20), which are typical attractants of Langerhans cells, by quantitative reverse transcriptase polymerase chain reaction. The expression of both chemokines, CCL2 and CCL20, were suppressed by treatment with cortisol (suppression by 38% and 48% compared to no stress hormone, respectively). We examined the possible regulation of the suppression by cortisol with plant extracts. The extracts of Ganoderma lucidum and Iris protected the suppression of the differentiation to CD39-positive cells and also the suppression of the gene expression of LC-chemoattractants. These results suggest that cortisol, which is either systemic or locally produced, blocks the supply of epidermal Langerhans cells at 2 steps, differentiation from the precursor and attraction to epidermis. The suppression is possibly blocked by some plant extracts.

Keywords: Langerhans cell, stress, CD39, chemokine

Procedia PDF Downloads 189

Authors: Tahir Iqbal, Mumtaz Ali, Syed Wajahat Kareem, Muhammad Harris

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Digital information is expanding in exponential order in our life. Information that is residing online and offline are stored in huge repositories relating to every aspect of our lives. Getting the required information is a task of retrieval systems. Content based image retrieval (CBIR) is a retrieval system that retrieves the required information from repositories on the basis of the contents of the image. Time is a critical factor in retrieval system and using indexed views with CBIR system improves the time efficiency of retrieved results.

Keywords: content based image retrieval (CBIR), indexed view, color, image retrieval, cross correlation

Procedia PDF Downloads 474

Authors: Mustafa Yıldız, Ali Ulvi Uzer, Murat Olgun

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In this study, geotechnical and mineralogical properties of gypsum containing clay basis which form the ground of Second Organized Industrial Zone in Konya province have been researched through comprehensive field and laboratory experiments. Although sufficient geotechnical research has not been performed yet, an intensive structuring in the region continues at present. The study area consists of mid-lake sediments formed by gypsum containing soft silt-clay basis which evolves to a large area. To determine the soil profile and geotechnical specifications; 18 drilling holes were opened and disturbed / undisturbed soil samples have been taken through shelby tubes within 1.5m intervals. Tests have been performed on these samples to designate the index and strength properties of soil. Besides, at all drilling holes Standart Penetration Tests have been done within 1.5m intervals. For the purpose of determining the mineralogical characteristics of the soil; all rock and X-RD analysis have been carried out on 6 samples which were taken from various depths through the soil profile. Strength and compressibility characteristics of the soil were defined with correlations using laboratory and field test results. Unconfined compressive strength, undrained cohesion, compression index varies between 16 kN/m2 and 405.4 kN/m2, 6.5 kN/m2 and 72 kN/m2, 0.066 and 0.864, respectively.

Keywords: Konya second organized industrial region, strength, compressibility, soft clay

Procedia PDF Downloads 311

Authors: Pranitha Janapatla, Venkata Suman Gontla

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The present paper investigates the effects of MHD, double dispersion and variable properties on mixed convection flow from a vertical surface in a power-law fluid saturated non-Darcy porous medium. The governing non-linear partial differential equations are reduced to a system of ordinary differential equations by using a special form of Lie group transformations viz. scaling group of transformations. These ordinary differential equations are solved numerically by using Shooting technique. The influence of relevant parameters on the non-dimensional velocity, temperature, concentration for pseudo-plastic fluid, Newtonian and dilatant fluid are discussed and displayed graphically. The behavior of heat and mass transfer coefficients are shown in tabular form. Comparisons with the published works are performed and are found to be in very good agreement. From this analysis, it is observed that an increase in variable viscosity causes to decrease in velocity profile and increase the temperature and concentration distributions. It is also concluded that increase in the solutal dispersion decreases the velocity and concentration but raises the temperature profile.

Keywords: power-law fluid, thermal conductivity, thermal dispersion, solutal dispersion, variable viscosity

Procedia PDF Downloads 239

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

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

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

Procedia PDF Downloads 161

Authors: Nelson Andres, Meeko C. Masangcap, John Denver D. Catapang

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Balanga City as, the heart of Bataan, is a growing City and is now at its fast pace of development. The growth of commerce in the city results to an increase in commuters who travel back and forth through the city, leading to congestions. Consequently, queuing of vehicles along national roads and even in the highways of the city have become a regular occurrence. This common scenario of commuters flocking the city, private and public vehicles going bumper to bumper, especially during the rush hours, greatly affect the flow of traffic vehicles and is now a burden not only to the commuters but also to the government who is trying to address this dilemma. Seeing these terrible events, the implementation of an elevated Automated Guide-way transit is seen as a possible solution to help in the decongestion of the affected parts of Balanga City.In response to the problem, the researchers identify if it is feasible to have an elevated guide-way transit in the vicinity of Sapphire Street in Balanga City, Bataan. Specifically, the study aims to determine who will be the riders based on the demographic profile, where the trip can be generated and distributed, the time when volume of people usually peaks and the estimated volume of passengers. Statistical analysis is applied to the data gathered to find out if there is an important relationship between the demographic profile of the respondents and their preference of having an elevated railway transit in the City of Balanga.

Keywords: ridership, AGT, railway, elevated track

Procedia PDF Downloads 85

Authors: A. Ramachandra Reddy, V. Murugan, Prema Kumari

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Residual solvents in Pharmaceutical samples are monitored using gas chromatography with headspace (GC-HS). Based on current regulatory and compendial requirements, measuring the residual solvents are mandatory for all release testing of active pharmaceutical ingredients (API). Generally, isopropyl alcohol is used as the residual solvent in proline and tryptophan; methanol in cysteine monohydrate hydrochloride, glycine, methionine and serine; ethanol in glycine and lysine monohydrate; acetic acid in methionine. In order to have a single method for determining these residual solvents (isopropyl alcohol, ethanol, methanol and acetic acid) in all these 7 amino acids a sensitive and simple method was developed by using gas chromatography headspace technique with flame ionization detection. During development, no reproducibility, retention time variation and bad peak shape of acetic acid peaks were identified due to the reaction of acetic acid with the stationary phase (cyanopropyl dimethyl polysiloxane phase) of column and dissociation of acetic acid with water (if diluent) while applying temperature gradient. Therefore, dimethyl sulfoxide was used as diluent to avoid these issues. But most the methods published for acetic acid quantification by GC-HS uses derivatisation technique to protect acetic acid. As per compendia, risk-based approach was selected as appropriate to determine the degree and extent of the validation process to assure the fitness of the procedure. Therefore, Total error concept was selected to validate the analytical procedure. An accuracy profile of ±40% was selected for lower level (quantitation limit level) and for other levels ±30% with 95% confidence interval (risk profile 5%). The method was developed using DB-Waxetr column manufactured by Agilent contains 530 µm internal diameter, thickness: 2.0 µm, and length: 30 m. A constant flow of 6.0 mL/min. with constant make up mode of Helium gas was selected as a carrier gas. The present method is simple, rapid, and accurate, which is suitable for rapid analysis of isopropyl alcohol, ethanol, methanol and acetic acid in amino acids. The range of the method for isopropyl alcohol is 50ppm to 200ppm, ethanol is 50ppm to 3000ppm, methanol is 50ppm to 400ppm and acetic acid 100ppm to 400ppm, which covers the specification limits provided in European pharmacopeia. The accuracy profile and risk profile generated as part of validation were found to be satisfactory. Therefore, this method can be used for testing of residual solvents in amino acids drug substances.

Keywords: amino acid, head space, gas chromatography, total error

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Authors: Dan Geneau, Mary Claire Geneau, Seth Lenetsky, Ming -Chang Tsai, Marc Klimstra

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Introduction— Horizontal force-velocity profiling (HFVP) involves modeling an athletes linear sprint kinematics to estimate valuable maximum force and velocity metrics. This approach to performance modeling has been used in field-based team sports and has recently been introduced to ice-hockey as a forward skating performance assessment. While preliminary data has been collected on ice, distance constraints of the on-ice test restrict the ability of the athletes to reach their maximal velocity which result in limits of the model to effectively estimate athlete performance. This is especially true of more elite athletes. This report explores whether athletes on-ice are able to reach a velocity plateau similar to what has been seen in overground trials. Fourteen male Major Junior ice-hockey players (BW= 83.87 +/- 7.30 kg, height = 188 ± 3.4cm cm, age = 18 ± 1.2 years n = 14) were recruited. For on-ice sprints, participants completed a standardized warm-up consisting of skating and dynamic stretching and a progression of three skating efforts from 50% to 95%. Following the warm-up, participants completed three on ice 45m sprints, with three minutes of rest in between each trial. For overground sprints, participants completed a similar dynamic warm-up to that of on-ice trials. Following the warm-up participants completed three 40m overground sprint trials. For each trial (on-ice and overground), radar was used to collect instantaneous velocity (Stalker ATS II, Texas, USA) aimed at the participant’s waist. Sprint velocities were modelled using custom Python (version 3.2) script using a mono-exponential function, similar to previous work. To determine if on-ice tirals were achieving a maximum velocity (plateau), minimum acceleration values of the modeled data at the end of the sprint were compared (using paired t-test) between on-ice and overground trials. Significant differences (P<0.001) between overground and on-ice minimum accelerations were observed. It was found that on-ice trials consistently reported higher final acceleration values, indicating a maximum maintained velocity (plateau) had not been reached. Based on these preliminary findings, it is suggested that reliable HFVP metrics cannot yet be collected from all ice-hockey populations using current methods. Elite male populations were not able to achieve a velocity plateau similar to what has been seen in overground trials, indicating the absence of a maximum velocity measure. With current velocity and acceleration modeling techniques, including a dependency of a velocity plateau, these results indicate the potential for error in on-ice HFVP measures. Therefore, these findings suggest that a greater on-ice sprint distance may be required or the need for other velocity modeling techniques, where maximal velocity is not required for a complete profile.   

Keywords: ice-hockey, sprint, skating, power

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Authors: Elena Maria Scalisi

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Currently, photodegradation by nanoparticles (NPs) is a common solution for wastewater treatment. Nanoparticles are efficient for removing organic and inorganic pollutants, heavy metals from wastewater and killing microorganisms through environmentally friendly. In this context, the major representative of photocatalytic technology for industrial wastewater treatment are TiO₂ nanoparticles (TiO₂-NPs). TiO₂-NPs have a strong catalytic activity that depends to their physicochemical properties. Thanks to their small size (between 1-100 nm), nanoparticles occupy less volume, then their surface area increases. The increase in the surface-to-volume ratio results in the increase of the particle surface energy, which improve their reactivity potential. However, these unique properties represent risks to the ecosystems and organisms when unintentionally TiO₂-NPs are release into the environment and absorbed by living organisms. Several studies confirm that there is a high level of interest concerning the safety of TiO₂-NPs in the aquatic environment, furthermore, ecotoxicological tools are useful to correctly evaluate their toxicity. In the current study, we aimed to characterize potential toxic effects of TiO₂-NP suspension to zebrafish during embryo-larval stages to evaluate parameters such as survival rates, malformation, hatching, the overall length of the larvae heartbeat, and biochemical biomarkers that reflect the acute toxicity and sublethal effects of TiO₂-NPs. Zebrafish embryos were exposed to titanium dioxide nanoparticles (TiO₂-NPs at 1mg/L, 2mg/L, and 4mg/L) from fertilization to the free swimming stage (144hpf). Every day, we recorded the toxicological endpoints, moreover, immunohistochemical analysis has been performed at the end of the exposure. In particular, we have evaluate the expression of the following biomarkers: Heat Shock Protein 70 (HSP70), Poly ADP-Ribose Polymerase-1 (PARP-1), Metallothioneins (MTs). Our results have shown that hatch ability, survival, and malformation rate were not affected by TiO₂ NPs at these exposure levels. However, TiO₂-NPs caused an increase of heartbeat and reduction of body length; at the same time, TiO₂-NPs have inducted the production of ROS and the expression of oxidative stress biomarkers HSP70 and PARP-1. Hight positivity for PARP-1 at all concentration tested was observed. As regards MT, positivity was found in the expression of this biomarker in the whole body of the embryo, with the exception of the end of the tail. Metallothioneins (MT) are biomarkers widely used in environmental monitoring programs for aquatic creatures. At the light of our results i.e. no death until the end of the experiment (144hpf), no malformation and expression of the biomarkers mentioned, it is evident that zebrafish larvae with their natural detoxification pathways are able to resist the presence of toxic substances and then they can tolerate the presence of metal concentrations. However, an excessive oxidative state can compromise cell function, therefore the uncontrolled release of nanoparticles into the environment is severe and must be constantly monitored.

Keywords: nanoparticles, embryo zebrafish, HSP70, PARP-1

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Authors: Rachel Koshy, Nazrila Hairizan Bt. Nasir, Samsiah Bt. Awang, Kamaliah Bt. Mohamad Noh

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Malaysia collaborated as a ‘trailblazer’ country with PHCPI (Primary Health Care Performance Initiative) to populate the Primary Health Care (PHC) Vital Signs Profile (VSP) for the country. The PHC VSP provides an innovative snapshot of the primary health care system's performance. Four domains were assessed: system financing, system capacity, system performance, and system equity, and completed in 2019. There were two phases using a mixed method study design. The first phase involved a quantitative study, utilising existing secondary data from national and international sources. In the case of unavailability of data for any indicators, comparable alternative indicators were used. The second phase was a mixed quantitative-qualitative approach to measure the functional capacity based on governance and leadership, population health needs, inputs, population health management, and facility organisation and management. PHC spending constituted 35% of overall health spending in Malaysia, with a per capita PHC spending of $152. The capacity domain was strong in the three subdomains of governance and leadership, information system, and funds management. The two subdomains of drugs & supplies and facility organisation & management had low scores, but the lowest score was in empanelment of the population under the population health management. The PHC system performed with an access index of 98%, quality index of 84%, and service coverage of 62%. In the equity domain, there was little fluctuation in the coverage of reproductive, maternal, newborn, and child health services by mother’s level of education and under-five child mortality between urban and rural areas. The public sector was stronger in the capacity domain as compared to the private sector. This is due to the different financing, organisational structures, and service delivery mechanism. The VSP has identified areas for improvement in the effort to provide high-quality PHC for the population. The gaps in PHC can be addressed through the system approach and the positioning of public and private primary health care delivery systems.

Keywords: primary health care, health system, system domains, vital signs profile

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Authors: Abhishek Kumar Sah, Preeti K. Suresh

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Effective drug delivery to the eye is a massive challenge, due to complicated physiological ocular barriers, rapid washout by tear and nasolachrymal drainage. Thus, most of the conventional ophthalmic formulations face the problem of low ocular bioavailability. Ophthalmic drug therapy can be improved by enhancing the precorneal drug retention along with improved drug penetration. The aim of the present investigation was to develop and evaluate a biodegradable polymer poly (D, L-lactide-co-glycolide) (PLGA) coated nanoparticulate carrier of loteprednol etabonate. PLGA nanoparticles were prepared by modified emulsification/solvent diffusion method using high-speed homogenizer followed by sonication. The nanoparticles were characterized for various parameters such as particle size, zeta potential, polydispersity index, X-ray powder diffraction (XRD), Transmission electron microscopy (TEM), in vitro drug release profile and stability. The prepared nanocarriers displayed mean particle size in the range of 271.7 to 424.4 nm, with zeta potential less than –10 mV. In vitro release in simulated tear fluid (STF) nanocarrier showed an extended release profile of loteprednol etabonate. TEM confirmed the spherical morphology and smooth surface of the particles. All the prepared formulations were found to be stable at varying temperatures.

Keywords: drug delivery, ocular delivery, polymeric nanoparticles, loteprednol etabonate

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Authors: W. S. Thulasitha, N. Umasuthan, G. I. Godahewa, Jehee Lee

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In fish, innate immune defense is the first immune response against microbial pathogens which consists of several antimicrobial components. Galectins are one of the carbohydrate binding lectins that have the ability to identify pathogen by recognition of pathogen associated molecular patterns. Galectins play a vital role in the regulation of innate and adaptive immune responses. Rock bream Oplegnathus fasciatus is one of the most important cultured species in Korea and Japan. Considering the losses due to microbial pathogens, present study was carried out to understand the molecular and functional characteristics of a galectin in normal and pathogenic conditions, which could help to establish an understanding about immunological components of rock bream. Complete cDNA of rock bream galectin like protein B (rbGal like B) was identified from the cDNA library, and the in silico analysis was carried out using bioinformatic tools. Genomic structure was derived from the BAC library by sequencing a specific clone and using Spidey. Full length of rbGal like B (contig14775) cDNA containing 517 nucleotides was identified from the cDNA library which comprised of 435 bp in the open reading frame encoding a deduced protein composed of 145 amino acids. The molecular mass of putative protein was predicted as 16.14 kDa with an isoelectric point of 8.55. A characteristic conserved galactose binding domain was located from 12 to 145 amino acids. Genomic structure of rbGal like B consisted of 4 exons and 3 introns. Moreover, pairwise alignment showed that rock bream rbGal like B shares highest similarity (95.9 %) and identity (91 %) with Takifugu rubripes galectin related protein B like and lowest similarity (55.5 %) and identity (32.4 %) with Homo sapiens. Multiple sequence alignment demonstrated that the galectin related protein B was conserved among vertebrates. A phylogenetic analysis revealed that rbGal like B protein clustered together with other fish homologs in fish clade. It showed closer evolutionary link with Takifugu rubripes. Tissue distribution and expression patterns of rbGal like B upon immune challenges were performed using qRT-PCR assays. Among all tested tissues, level of rbGal like B expression was significantly high in gill tissue followed by kidney, intestine, heart and spleen. Upon immune challenges, it showed an up-regulated pattern of expression with Edwardsiella tarda, rock bream irido virus and poly I:C up to 6 h post injection and up to 24 h with LPS. However, In the presence of Streptococcus iniae rbGal like B showed an up and down pattern of expression with the peak at 6 - 12 h. Results from the present study revealed the phylogenetic position and role of rbGal like B in response to microbial infection in rock bream.

Keywords: galectin like protein B, immune response, Oplegnathus fasciatus, molecular characterization

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Authors: Bushra Nabi, Saleha Rehman, Sanjula Baboota, Javed Ali

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Aim: The objective of research undertaken is analytical method validation (HPLC method) of an anti-HIV drug Elvitegravir (EVG). Additionally carrying out the forced degradation studies of the drug under different stress conditions to determine its stability. It is envisaged in order to determine the suitable technique for drug estimation, which would be employed in further research. Furthermore, comparative pharmacokinetic profile of the drug from lipid architectonics and drug suspension would be obtained post oral administration. Method: Lipid Architectonics (LA) of EVR was formulated using probe sonication technique and optimized using QbD (Box-Behnken design). For the estimation of drug during further analysis HPLC method has been validation on the parameters (Linearity, Precision, Accuracy, Robustness) and Limit of Detection (LOD) and Limit of Quantification (LOQ) has been determined. Furthermore, HPLC quantification of forced degradation studies was carried out under different stress conditions (acid induced, base induced, oxidative, photolytic and thermal). For pharmacokinetic (PK) study, Albino Wistar rats were used weighing between 200-250g. Different formulations were given per oral route, and blood was collected at designated time intervals. A plasma concentration profile over time was plotted from which the following parameters were determined:

Keywords: AIDS, Elvitegravir, HPLC, nanostructured lipid carriers, pharmacokinetics

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Authors: Xiang Jinhua, Mclinden James, Chang Qing, Fosdick Micaela, Ploss Alexander, Bhattarai Nirjal, Houtman Jon, Stapleton Jack

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Background: Clinical outcomes differ among dengue virus (DENV) serotypes though few serotype-specific differences are identified. We previously found that ZIKV and DENV-2 envelope proteins do not inhibit T cell receptor (TCR) signaling. Here, we investigated the effect of DENV-1-4 infection and env expression on T cell functions. Methods: DENV-1 though 4 were added to PBMCs or Jurkat T cells prior to TCR stimulation. Signaling was measured by IL-2 release. The effect of DENV env (1-4) expression in primary and Jurkat T cells on TCR was measured. DENV env regions required for TCR inhibition were mapped by chimera, deletions, and point mutagenesis. Results were confirmed by reverse genetics using replication competent DENV generated by CPER. Results: DENV-1-4 caused abortive infection in T cells, yet DENV-1 and -4 inhibited TCR signaling measured by IL-2 release in primary and transformed T cells while DENV-2 and -3 did not. This was not due to differences in binding, entry or RNA production. DENV-1 and -4 env expression in Jurkat or exposure in primary T cells recapitulated TCR inhibition. The Env sequences in volved were mapped and mutation of Env a.a V55 to the T present in DENV-2 and -3 abolished TCR inhibition in replicating viruses. Substituting the DENV-1 55V into DENV 2, 3 led to partial TCR inhibition; however, addition T66S into the V55T DENV-2 and -3 rescued the TCR inhibition found in DENV-1 and -4. Preliminary data suggest that the envelope substitutions may reduce replication kinetics in different mammalian or insect cells, and this is under further study. Conclusions: Epidemiological data suggest that DENV 2 and 3 are more often associated with severe dengue including hemorrhagic fever and shock. Since DENV-1 and -4 interfere with TCR, it is possible that this TCR effect blunts host immunologic responses during infection, mitigating immune-mediated pathogenic effects of DENV. Recombinant viruses demonstrate that DENV1 V55T substitution is sufficient to remove the TCR inhibitory phenotype, and that changing DENV-2, -3 aa 55 and 66 to that seen in DENV-1 mimics TCR inhibition observed in DENV-1 and -4. These findings may provide an approach to safer live-attenuated DENV vaccines.

Keywords: dengue Viruses, TCR signaling, CPER, dengue viral envelope protein

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Authors: Syeda Kiran Shahzadi, Nasir Mahmood, Muhammad Abdul Qadir

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In the current research, three recombinant human interferon alpha-2b proteins (two modified and one normal form) were produced and Pegylated with an aim to produce more effective drugs against viral infections and cancers. The modified recombinant human interferon alpha-2b proteins were produced by site-directed modifications of interferon alpha 2b gene, targeting the amino acids at positions ‘R23’ and ‘H34’. The resulting chemically modified and unmodified forms of human interferon alpha 2b were conjugated with methoxy-polyethylene glycol propanealdehyde (400 KDa) and methoxy-polyethylene glycol succinimidyl succinate (400 KDa). Pegylation of normal and modified forms of Interferon alpha-2b prolong their release time and enhance their efficacy. The conjugation of PEG with modified and unmodified human interferon alpha 2b protein drugs was also characterized with 1H-NMR, HPLC, and SDS-PAGE. Antiproliferative assays of modified and unmodified forms of drugs were performed in cell based bioassays using MDBK cell lines. The results indicated that experimentally produced recombinant human interferon alpha-2b proteins were biologically active and resulted in significant inhibition of cell growth.

Keywords: protein refolding, antiproliferative activities, biomedical applications, human interferon alpha-2b, pegylation, mPEG-propionaldehyde, site directed mutagenesis, E. coli expression

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Authors: Michael R. Shurin, Galina Shurin, Vladimir A. Kirichenko

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Background. Visibility of hepatic malignancies is poor on non-contrast imaging for daily verification of liver malignancies prior to radiation therapy on MRI-guided Linear Accelerators (MR-Linac). Ferumoxytol® (Feraheme, AMAG Pharmaceuticals, Waltham, MA) is a SPION agent that is increasingly utilized off-label as hepatic MRI contrast. This agent has the advantage of providing a functional assessment of the liver based upon its uptake by hepatic Kupffer cells proportionate to vascular perfusion, resulting in strong T1, T2 and T2* relaxation effects and enhanced contrast of malignant tumors, which lack Kupffer cells. The latter characteristic has been recently utilized for MRI-guided radiotherapy planning with precision targeting of liver malignancies. However potential radiotoxicity of SPION has never been addressed for its safe use as an MRI-contrast agent during liver radiotherapy on MRI-Linac. This study defines the radiomodulating properties of SPIONs in vitro on human monocyte and macrophage cell lines exposed to 60Go gamma-rays within clinical radiotherapy dose range. Methods. Human monocyte and macrophages cell line in cultures were loaded with a clinically relevant concentration of Ferumoxytol (30µg/ml) for 2 and 24 h and irradiated to 3Gy, 5Gy and 10Gy. Cells were washed and cultured for additional 24 and 48 h prior to assessing their phenotypic activation by flow cytometry and function, including viability (Annexin V/PI assay), proliferation (MTT assay) and cytokine expression (Luminex assay). Results. Our results reveled that SPION affected both human monocytes and macrophages in vitro. Specifically, iron oxide nanoparticles decreased radiation-induced apoptosis and prevented radiation-induced inhibition of human monocyte proliferative activity. Furthermore, Ferumoxytol protected monocytes from radiation-induced modulation of phenotype. For instance, while irradiation decreased polarization of monocytes to CD11b+CD14+ and CD11bnegCD14neg phenotype, Ferumoxytol prevented these effects. In macrophages, Ferumoxytol counteracted the ability of radiation to up-regulate cell polarization to CD11b+CD14+ phenotype and prevented radiation-induced down-regulation of expression of HLA-DR and CD86 molecules. Finally, Ferumoxytol uptake by human monocytes down-regulated expression of pro-inflammatory chemokines MIP-1α (Macrophage inflammatory protein 1α), MIP-1β (CCL4) and RANTES (CCL5). In macrophages, Ferumoxytol reversed the expression of IL-1RA, IL-8, IP-10 (CXCL10) and TNF-α, and up-regulates expression of MCP-1 (CCL2) and MIP-1α in irradiated macrophages. Conclusion. SPION agent Ferumoxytol increases resistance of human monocytes to radiation-induced cell death in vitro and supports anti-inflammatory phenotype of human macrophages under radiation. The effect is radiation dose-dependent and depends on the duration of Feraheme uptake. This study also finds strong evidence that SPIONs reversed the effect of radiation on the expression of pro-inflammatory cytokines involved in initiation and development of radiation-induced liver damage. Correlative translational work at our institution will directly assess the cyto-protective effects of Ferumoxytol on human Kupfer cells in vitro and ex vivo analysis of explanted liver specimens in a subset of patients receiving Feraheme-enhanced MRI-guided radiotherapy to the primary liver tumors as a bridge to liver transplant.

Keywords: superparamagnetic iron oxide nanoparticles, radioprotection, magnetic resonance imaging, liver

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Authors: Liu Xiaohan

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Epstein–Barr virus (EBV) is a human herpesvirus and is closely related to many malignancies of lymphocyte and epithelial origins, such as gastric cancer, Burkitt’s lymphoma, and nasopharyngeal carcinoma (NPC). NPC is a malignant epithelial tumor which is 100% associated with EBV latent infection. Most of the NPC cases are densely populated in southern China, especially in Guangdong and Hong Kong. To our knowledge, overexpression of pro-inflammatory cytokines may result in a loss of balance of the immune system and cause damage to human bodies. Interleukin-6 (IL6) is a pro-inflammatory cytokine which plays an important role in tumor progression. In addition, gene expression is regulated by both transcriptional and post-transcriptional pathways, while post-transcriptional regulation is an important mechanism to modulate the mature mRNA level in mammalian cells. AU-rich element binding factor 1 (AUF1)/heterogeneous nuclear RNP D (hnRNP D) is known for its function in destabilizing mRNAs, including cytokines and cell cycle regulators. Previous studies have found that overexpression of hnRNP D would lead to tumorigenesis. In this project, our aim is to determine the role played by hnRNP D in EBV-infected cells and how our anti-EBV agents can affect the function of hnRNP D. The results of this study will provide a new insight into how the pro-inflammatory cytokine expression can be regulated by EBV.

Keywords: interleukin 6 (IL6), epstein-barr virus (EBV), nasopharyngeal carcinoma (NPC, epstein-barr nuclear antigen-1 (EBNA1)

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Authors: Sarwan Dhir, Suma Basak, Dipika Parajulee

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Alfalfa is renowned for its nutritional and biopharmaceutical value as a perennial forage legume. However, establishing a rapid plant regeneration protocol using somatic embryogenesis and efficient transformation frequency are the crucial prerequisites for gene editing in alfalfa. This study was undertaken to establish and improve the protocol for somatic embryogenesis and subsequent plant regeneration. The experiments were conducted in response to natural sensitivity using various antibiotics such as cefotaxime, carbenicillin, gentamycin, hygromycin, and kanamycin. Using 3-week-old leaf tissue, somatic embryogenesis was initiated on Gamborg’s B5 basal (B5H) medium supplemented with 3% maltose, 0.9µM Kinetin, and 4.5µM 2,4-D. Embryogenic callus (EC) obtained from the B5H medium exhibited a high rate of somatic embryo formation (97.9%) after 3 weeks when the cultures were placed in the dark. Different developmental stages of somatic embryos and cotyledonary stages were then transferred to Murashige and Skoog’s (MS) basal medium under light, resulting in a 94% regeneration rate of plantlets. Our results indicate that leaf segments can grow (tolerate) up to 450 mg/L of cefotaxime and 400 mg/L of carbenicillin in the culture medium. However, the survival threshold for hygromycin at 12.5 mg/L, kanamycin at 250 mg/L, gentamycin at 50 mg/L, and timentin (300 mg/L). The experiment to improve the protocol for achieving efficient transient gene expression in alfalfa through genetic transformation with the Agrobacterium tumefaciens pCAMBIA1304 vector was also conducted. The vector contains two reporter genes such as β-glucuronidase (GUS) and green fluorescent protein (GFP), along with a selectable hygromycin B phosphotransferase gene (HPT), all driven under the CaMV 35s promoter. Various transformation parameters were optimized using 3-week-old in vitro-grown plantlets. The different parameters such as types of explant, leaf ages, preculture days, segment sizes, wounding types, bacterial concentrations, infection periods, co-cultivation periods, different concentrations of acetosyringone, silver nitrate, and calcium chloride were optimized for transient gene expression. The transient gene expression was confirmed via histochemical GUS and GFP visualization under fluorescent microscopy. The data were analyzed based on the semi-quantitative observation of the percentage and number of blue GUS spots on different days of agro-infection. The highest percentage of GUS positivity (76.2%) was observed in 3-week-old leaf segments wounded using a scalpel blade of 11 size- after 3 days of post-incubation at a bacterial concentration of 0.6, with 2 days of preculture, 30 min of bacterial-leaf segment co-cultivation, with the addition of 150 µM acetosyringone, 4 mM calcium chloride, and 75 µM silver nitrate. Our results suggest that various factors influence T-DNA delivery in the Agrobacterium-mediated transformation of alfalfa. The stable gene expression in the putative transgenic tissue was confirmed using PCR amplification of both marker genes, indicating that gene expression in explants was not solely due to Agrobacterium, but also from transformed cells. The improved protocol could be used for generating transgenic alfalfa plants using genome editing techniques such as CRISPR/Cas9.

Keywords: Medicago sativa l. (Alfalfa), agrobacterium tumefaciens, β-glucuronidase, green fluorescent protein, transient gene

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Authors: Chanh Nghia Nguyen, Yu Kurokawa, Hirotsugu Inoue

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The surface roughness is an important parameter for evaluating the quality of material surfaces since it affects functions and performance of industrial components. Although stylus and optical techniques are commonly used for measuring the surface roughness, they are applicable only to accessible surfaces. In practice, surface roughness measurement from the back side is sometimes demanded, for example, in inspection of safety-critical parts such as inner surface of pipes. However, little attention has been paid to the measurement of back surface roughness so far. Since back surface is usually inaccessible by stylus or optical techniques, ultrasonic technique is one of the most effective among others. In this research, an ultrasonic pulse-echo technique is considered for evaluating the pitch and the height of back surface having periodic triangular profile as a very first step. The pitch of the surface profile is measured by applying the diffraction grating theory for oblique incidence; then the height is evaluated by numerical analysis based on the Kirchhoff theory for normal incidence. The validity of the proposed method was verified by both numerical simulation and experiment. It was confirmed that the pitch is accurately measured in most cases. The height was also evaluated with good accuracy when it is smaller than a half of the pitch because of the approximation in the Kirchhoff theory.

Keywords: back side, inaccessible surface, periodic roughness, pulse-echo technique, ultrasonic NDE

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Authors: Varsha Prasad, S. Sandya

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Cosmic Ray effects in microelectronics such as single event effect (SET) and total dose ionization (TID) have been of major concern in space electronics since 1970. Advanced CMOS technologies have demonstrated reduced sensitivity to TID effect. However, charge pump Phase Locked Loop is very much vulnerable to single event transient effect. This paper presents an SET analysis model, where the SET is modeled as a double exponential pulse. The time domain analysis reveals that the settling time of the voltage controlled oscillator (VCO) depends on the SET pulse strength, setting the time constant and the damping factor. The analysis of the proposed SET analysis model is confirmed by the simulation results.

Keywords: charge pump, phase locked loop, SET, VCO

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Authors: Bianca Peterson, Tomasz J. Sańko, Carlos C. Bezuidenhout, Johnnie Van Den Berg

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Busseola fusca (Fuller) (Lepidoptera: Noctuidae), the maize stem borer, is a major pest in sub-Saharan Africa. It causes economic damage to maize and sorghum crops and has evolved non-recessive resistance to genetically modified (GM) maize expressing the Cry1Ab insecticidal toxin. Since B. fusca is a non-model organism, very little genomic information is publicly available, and is limited to some cytochrome c oxidase I, cytochrome b, and microsatellite data. The biology of B. fusca is well-described, but still poorly understood. This, in combination with its larval-specific behavior, may pose problems for limiting the spread of current resistant B. fusca populations or preventing resistance evolution in other susceptible populations. As part of on-going research into resistance evolution, B. fusca larvae were collected from Bt and non-Bt maize in South Africa, followed by RNA isolation (15 specimens) and sequencing on the Illumina HiSeq 2500 platform. Quality of reads was assessed with FastQC, after which Trimmomatic was used to trim adapters and remove low quality, short reads. Trinity was used for the de novo assembly, whereas TransRate was used for assembly quality assessment. Transcript identification employed BLAST (BLASTn, BLASTp, and tBLASTx comparisons), for which two libraries (nucleotide and protein) were created from 3.27 million lepidopteran sequences. Several transcripts that have previously been implicated in Cry toxin resistance was identified for B. fusca. These included aminopeptidase N, cadherin, alkaline phosphatase, ATP-binding cassette transporter proteins, and mitogen-activated protein kinase. MEGA7 was used to align these transcripts to reference sequences from Lepidoptera to detect mutations that might potentially be contributing to Cry toxin resistance in this pest. RSEM and Bioconductor were used to perform differential gene expression analysis on groups of B. fusca larvae challenged and unchallenged with the Cry1Ab toxin. Pairwise expression comparisons of transcripts that were at least 16-fold expressed at a false-discovery corrected statistical significance (p) ≤ 0.001 were extracted and visualized in a hierarchically clustered heatmap using R. A total of 329,194 transcripts with an N50 of 1,019 bp were generated from the over 167.5 million high-quality paired-end reads. Furthermore, 110 transcripts were over 10 kbp long, of which the largest one was 29,395 bp. BLAST comparisons resulted in identification of 157,099 (47.72%) transcripts, among which only 3,718 (2.37%) were identified as Cry toxin receptors from lepidopteran insects. According to transcript expression profiles, transcripts were grouped into three subclusters according to the similarity of their expression patterns. Several immune-related transcripts (pathogen recognition receptors, antimicrobial peptides, and inhibitors) were up-regulated in the larvae feeding on Bt maize, indicating an enhanced immune status in response to toxin exposure. Above all, extremely up-regulated arylphorin genes suggest that enhanced epithelial healing is one of the resistance mechanisms employed by B. fusca larvae against the Cry1Ab toxin. This study is the first to provide a resource base and some insights into a potential mechanism of Cry1Ab toxin resistance in B. fusca. Transcriptomic data generated in this study allows identification of genes that can be targeted by biotechnological improvements of GM crops.

Keywords: epithelial healing, Lepidoptera, resistance, transcriptome

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