Search results for: Cancer Stem Cells

Authors: Silvia Portarena, Chiara Anselmi, Chiara Baldacchini, Enrico Brugnoli

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Extra virgin olive oil (EVOO) is a complex matrix mainly composed by fatty acid and other minor compounds, among which carotenoids are well known for their antioxidative function that is a key mechanism of protection against cancer, cardiovascular diseases, and macular degeneration in humans. EVOO composition in terms of such constituents is generally the result of a complex combination of genetic, agronomical and environmental factors. To selectively improve the quality of EVOOs, the role of each factor on its biochemical composition need to be investigated. By selecting fruits from four different cultivars similarly grown and harvested, it was demonstrated that Raman spectroscopy, combined with chemometric analysis, is able to discriminate the different cultivars, also as a function of the harvest date, based on the relative content and composition of fatty acid and carotenoids. In particular, a correct classification up to 94.4% of samples, according to the cultivar and the maturation stage, was obtained. Moreover, by using gas chromatography and high-performance liquid chromatography as reference techniques, the Raman spectral features further allowed to build models, based on partial least squares regression, that were able to predict the relative amount of the main fatty acids and the main carotenoids in EVOO, with high coefficients of determination. Besides genetic factors, climatic parameters, such as light exposition, distance from the sea, temperature, and amount of precipitations could have a strong influence on EVOO composition of both major and minor compounds. This suggests that the Raman spectra could act as a specific fingerprint for the geographical discrimination and authentication of EVOO. To understand the influence of environment on EVOO Raman spectra, samples from seven regions along the Italian coasts were selected and analyzed. In particular, it was used a dual approach combining Raman spectroscopy and isotope ratio mass spectrometry (IRMS) with principal component and linear discriminant analysis. A correct classification of 82% EVOO based on their regional geographical origin was obtained. Raman spectra were obtained by Super Labram spectrometer equipped with an Argon laser (514.5 nm wavelenght). Analyses of stable isotope content ratio were performed using an isotope ratio mass spectrometer connected to an elemental analyzer and to a pyrolysis system. These studies demonstrate that RR spectroscopy is a valuable and useful technique for the analysis of EVOO. In combination with statistical analysis, it makes possible the assessment of specific samples’ content and allows for classifying oils according to their geographical and varietal origin.

Keywords: authentication, chemometrics, olive oil, raman spectroscopy

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Authors: Deniz Erçelen, Özlem Selcuk Bozkurt

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The course of daily human life calls for the need for memories and remembering the time and place for certain events. Recalling memories takes up a substantial amount of time for an individual. Unfortunately, scientists lack the proper technology to fully understand and observe different brain regions that interact to form or retrieve memories. The hippocampus, a complex brain structure located in the temporal lobe, plays a crucial role in memory. The hippocampus forms memories as well as allows the brain to retrieve them by ensuring that neurons fire together. This process is called “neural synchronization.” Sadly, the hippocampus is known to deteriorate often with age. Proteins and hormones, which repair and protect cells in the brain, typically decline as the age of an individual increase. With the deterioration of the hippocampus, an individual becomes more prone to memory loss. Many memory loss starts off as mild but may evolve into serious medical conditions such as dementia and Alzheimer’s disease. In their quest to fully comprehend how memories work, scientists have created many different kinds of technology that are used to examine the brain and neural pathways. For instance, Magnetic Resonance Imaging - or MRI- is used to collect detailed images of an individual's brain anatomy. In order to monitor and analyze brain functions, a different version of this machine called Functional Magnetic Resonance Imaging - or fMRI- is used. The fMRI is a neuroimaging procedure that is conducted when the target brain regions are active. It measures brain activity by detecting changes in blood flow associated with neural activity. Neurons need more oxygen when they are active. The fMRI measures the change in magnetization between blood which is oxygen-rich and oxygen-poor. This way, there is a detectable difference across brain regions, and scientists can monitor them. Electroencephalography - or EEG - is also a significant way to monitor the human brain. The EEG is more versatile and cost-efficient than an fMRI. An EEG measures electrical activity which has been generated by the numerous cortical layers of the brain. EEG allows scientists to be able to record brain processes that occur after external stimuli. EEGs have a very high temporal resolution. This quality makes it possible to measure synchronized neural activity and almost precisely track the contents of short-term memory. Science has come a long way in monitoring memories using these kinds of devices, which have resulted in the inspections of neurons and neural pathways becoming more intense and detailed.

Keywords: brain, EEG, fMRI, hippocampus, memories, neural pathways, neurons

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Authors: Deni̇z Ezgi̇ Gülmez, Mesut Kirca

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Nanoporous (np) metals have attracted considerable attention owing to their cellular morphological features at atomistic scale which yield ultra-high specific surface area awarding a great potential to be employed in diverse applications such as catalytic, electrocatalytic, sensing, mechanical and optical. As one of the carbon based nanostructures, fullerenes are also another type of outstanding nanomaterials that have been extensively investigated due to their remarkable chemical, mechanical and optical properties. In this study, the idea of improving the mechanical behavior of nanoporous metals by inclusion of the fullerenes, which offers a new metal-carbon nanocomposite material, is examined and discussed. With this motivation, tensile mechanical behavior of nanoporous metals reinforced with carbon fullerenes is investigated by classical molecular dynamics (MD) simulations. Atomistic models of the nanoporous metals with ultrathin ligaments are obtained through a stochastic process simply based on the intersection of spherical volumes which has been used previously in literature. According to this technique, the atoms within the ensemble of intersecting spherical volumes is removed from the pristine solid block of the selected metal, which results in porous structures with spherical cells. Following this, fullerene units are added into the cellular voids to obtain final atomistic configurations for the numerical tensile tests. Several numerical specimens are prepared with different number of fullerenes per cell and with varied fullerene sizes. LAMMPS code was used to perform classical MD simulations to conduct uniaxial tension experiments on np models filled by fullerenes. The interactions between the metal atoms are modeled by using embedded atomic method (EAM) while adaptive intermolecular reactive empirical bond order (AIREBO) potential is employed for the interaction of carbon atoms. Furthermore, atomic interactions between the metal and carbon atoms are represented by Lennard-Jones potential with appropriate parameters. In conclusion, the ultimate goal of the study is to present the effects of fullerenes embedded into the cellular structure of np metals on the tensile response of the porous metals. The results are believed to be informative and instructive for the experimentalists to synthesize hybrid nanoporous materials with improved properties and multifunctional characteristics.

Keywords: fullerene, intersecting spheres, molecular dynamic, nanoporous metals

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Authors: Aneta Maćczak, Bożena Bukowska, Jaromir Michałowicz

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Bisphenols are one of the most widely used chemical compounds worldwide. They are used in the manufacturing of polycarbonates, epoxy resins and thermal paper which are applied in plastic containers, bottles, cans, newspapers, receipt and other products. Among these compounds, bisphenol A (BPA) is produced in the highest amounts. There are concerns about endocrine impact of BPA and its other toxic effects including hepatotoxicity, neurotoxicity and carcinogenicity on human organism. Moreover, BPA is supposed to increase the incidence the obesity, diabetes and heart disease. For this reason the use of BPA in the production of plastic infant feeding bottles and some other consumers products has been restricted in the European Union and the United States. Nowadays, BPA analogues like bisphenol F (BPF) and bisphenol S (BPS) have been developed as alternative compounds. The replacement of BPA with other bisphenols contributed to the increase of the exposure of human population to these substances. Toxicological studies have mainly focused on BPA. In opposite, a small number of studies concerning toxic effects of BPA analogues have been realized, which makes impossible to state whether those substituents are safe for human health. Up to now, the mechanism of bisphenols action on the erythrocytes has not been elucidated. That is why, the aim of this study was to assess the effect of BPA and its selected analogues such as BPF and BPS on the activity of antioxidant enzymes, i.e. catalase (EC 1.11.1.6.), glutathione peroxidase (E.C.1.11.1.9) and superoxide dismutase (EC.1.15.1.1) in human erythrocytes. Red blood cells in respect to their function (transport of oxygen) and very well developed enzymatic and non-enzymatic antioxidative system, are useful cellular model to assess changes in redox balance. Erythrocytes were incubated with BPA, BPF and BPS in the concentration ranging from 0.5 to 100 µg/ml for 24 h. The activity of catalase was determined by the method of Aebi (1984). The activity of glutathione peroxidase was measured according to the method described by Rice-Evans et al. (1991), while the activity of superoxide dismutase (EC.1.15.1.1) was determined by the method of Misra and Fridovich (1972). The results showed that BPA and BPF caused changes in the antioxidative enzymes activities. BPA decreased the activity of examined enzymes in the concentration of 100 µg/ml. We also noted that BPF decreased the activity of catalase (5-100 µg/ml), glutathione peroxidase (50-100 µg/ml) and superoxide dismutase (25-100 µg/ml), while BPS did not cause statistically significant changes in investigated parameters. The obtained results suggest that BPA and BPF disrupt redox balance in human erythrocytes but the observed changes may occur in human organism only during occupational or subacute exposure to these substances.

Keywords: antioxidant enzymes, bisphenol A, bisphenol a analogues, human erythrocytes

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Authors: Samantha A. Cintron, Janet Pierce, Mihaela E. Sardiu, Diane Mahoney, Jill Peltzer, Bhanu Gupta, Qiuhua Shen

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High output heart failure (HOHF) is characterized a high output state resulting from an underlying disease process and is commonly caused by obesity. As obesity levels increase, more individuals will be at risk for obesity-related HOHF. However, the underlying pathophysiologic mechanisms of obesity-related HOHF are not well understood and need further research. The aim of the study was to describe the differences in leukocyte transcriptomes of morbidly obese patients with HOHF and those with non-HOHF. In this cross-sectional study, the study team collected blood samples, demographics, and clinical data of six patients with morbid obesity and HOHF and six patients with morbid obesity and non-HOHF. The study team isolated the peripheral blood leukocyte RNA and applied stranded total RNA sequencing. Differential gene expression was calculated, and Ingenuity Pathway Analysis software was used to interpret the canonical pathways, functional changes, upstream regulators, and mechanistic and causal networks that were associated with the significantly different leukocyte transcriptomes. The study team identified 116 differentially expressed genes; 114 were upregulated, and 2 were downregulated in the HOHF group (Benjamini-Hochberg adjusted p-value ≤ 0.05 and log2(fold-change) of ±1). The differentially expressed genes were involved with cell proliferation, mitochondrial function, erythropoiesis, erythrocyte stability, and apoptosis. The top upregulated canonical pathways associated with differentially expressed genes were autophagy, adenosine monophosphate-activated protein kinase signaling, and senescence pathways. Upstream regulator GATA Binding Protein 1 (GATA1) and a network associated with nuclear factor kappa-light chain-enhancer of activated B cells (NF-kB) were also identified based on the different leukocyte transcriptomes of morbidly obese patients with HOHF and non-HOHF. To the author’s best knowledge, this is the first study that reported the differential gene expression in patients with obesity-related HOHF and demonstrated the unique pathophysiologic mechanisms underlying the disease. Further research is needed to determine the role of cellular function and maintenance, inflammation, and iron homeostasis in obesity-related HOHF.

Keywords: cardiac output, heart failure, obesity, transcriptomics

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Authors: Saeed Rezaeian, M. Rezaee Boroon

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Nitrogen accumulates in soils during the process of fertilizer addition to promote the plant growth. When the organic matter decomposes, the form of available nitrogen produced is in the form of nitrate, which is highly mobile. The most significant health effect of nitrate ingestion is methemoglobinemia in infants under six months of age (blue baby syndrome). The mobile nutrients, like nitrate nitrogen, are not stored in the soil as the available forms for the long periods and in large amounts. It depends on the needs for the crops such as vegetables. On the other hand, the vegetables will compete actively for nitrate nitrogen as a mobile nutrient and water. The mobile nutrients must be shared. The fewer the plants, the larger this share is for each plant. Also, this nitrate nitrogen is poisonous for the people who use these vegetables. Nitrate is converted to nitrite by the existing bacteria in the stomach and the Gastro-Intestinal (GI) tract. When nitrite is entered into the blood cells, it converts the hemoglobin to methemoglobin, which causes the anoxemia and cyanosis. The increasing use of pesticides and chemical fertilizers, especially the fertilizers with nitrates compounds, which have been common for the increased production of agricultural crops, has caused the nitrate pollution in the (soil, water, and environment). They have caused a lot of damage to humans and animals. In this research, the nitrate accumulation in different kind of vegetables such as; green pepper, tomatoes, egg plants, watermelon, cucumber, and red pepper were observed in the suburbs of Mashhad, Neisabour, and Sabzevar cities. In some of these cities, the information forms of agronomical practices collected were such as; different vegetable crops fertilizer recommendations, varieties, pesticides, irrigation schedules, etc., which were filled out by some of our colleagues in the research areas mentioned above. Analysis of the samples was sent to the soil and water laboratory in our department in Mashhad. The final results from the chemical analysis of samples showed that the mean levels of nitrates from the samples of the fruit crops in the mentioned cities above were all lower than the critical levels. These fruit crop samples were in the order of: 35.91, 8.47, 24.81, 6.03, 46.43, 2.06 mg/kg dry matter, for the following crops such as; tomato, cucumber, eggplant, watermelon, green pepper, and red pepper. Even though, this study was conducted with limited samples and by considering the mean levels, the use of these crops from the nutritional point of view will not cause the poisoning of humans.

Keywords: environmental pollution, human health, nitrate accumulations, nitrate fertilizers

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Authors: Malahat Rezaee, Mahiran Basri, Abu Bakar Salleh, Raja Noor Zaliha Raja Abdul Rahman

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Sodium diclofenac is one of the most commonly used drugs of nonsteroidal anti-inflammatory drugs (NSAIDs). It is especially effective in the controlling the severe conditions of inflammation and pain, musculoskeletal disorders, arthritis, and dysmenorrhea. Formulation as nanoemulsions is one of the nanoscience approaches that has been progressively considered in pharmaceutical science for transdermal delivery of the drug. Nanoemulsions are a type of emulsion with particle sizes ranging from 20 nm to 200 nm. An emulsion is formed by the dispersion of one liquid, usually the oil phase in another immiscible liquid, water phase that is stabilized using the surfactant. Palm kernel oil esters (PKOEs), in comparison to other oils, contain higher amounts of shorter chain esters, which suitable to be applied in micro and nanoemulsion systems as a carrier for actives, with excellent wetting behavior without the oily feeling. This research aimed to study the effect of terpene type and concentration on sodium diclofenac permeation from palm kernel oil esters nanoemulsions and physicochemical properties of the nanoemulsions systems. The effect of various terpenes of geraniol, menthone, menthol, cineol and nerolidol at different concentrations of 0.5, 1.0, 2.0, and 4.0% on permeation of sodium diclofenac were evaluated using Franz diffusion cells and rat skin as permeation membrane. The results of this part demonstrated that all terpenes showed promoting effect on sodium diclofenac penetration. However, menthol and menthone at all concentrations showed significant effects (<0.05) on drug permeation. The most outstanding terpene was menthol with the most significant effect for skin permeability of sodium diclofenac. The effect of terpenes on physicochemical properties of nanoemulsion systems was investigated on the parameters of particle size, zeta potential, pH, viscosity and electrical conductivity. The result showed that all terpenes had the significant effect on particle size and non-significant effects on the zeta potential of the nanoemulsion systems. The effect of terpenes was significant on pH, excluding the menthone at concentrations of 0.5 and 1.0%, and cineol and nerolidol at the concentration of 2.0%. Terpenes also had significant effect on viscosity of nanoemulsions exception of menthone and cineol at the concentration of 0.5%. The result of conductivity measurements showed that all terpenes at all concentration except cineol at the concentration of 0.5% represented significant effect on electrical conductivity.

Keywords: nanoemulsions, palm kernel oil esters, sodium diclofenac, terpenes, skin permeation

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Authors: Ieva Bruzauskaite, Jovile Raudoniute, Karina Poliakovaite, Danguole Zabulyte, Daiva Bironaite, Ruta Aldonyte

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Introduction: Air pollution is a growing global problem which has been shown to be responsible for various adverse health outcomes. Immunotoxicity, such as dysregulated inflammation, has been proposed as one of the main mechanisms in air pollution-associated diseases. Chronic obstructive pulmonary disease (COPD) is among major morbidity and mortality causes worldwide and is characterized by persistent airflow limitation caused by the small airways disease (obstructive bronchiolitis) and irreversible parenchymal destruction (emphysema). Exact pathways explaining the air pollution induced and mediated disease states are still not clear. However, modern societies understand dangers of polluted air, seek to mitigate such effects and are in need for reliable biomarkers of air pollution. We hypothesise that post-translational modifications of structural proteins, e.g. citrullination, might be a good candidate biomarker. Thus, we have designed this study, where mice were exposed to diesel exhaust and the ongoing protein modifications and inflammation in lungs and other tissues were assessed. Materials And Methods: To assess the effects of diesel exhaust a in vivo study was designed. Mice (n=10) were subjected to everyday 2-hour exposure to diesel exhaust for 14 days. Control mice were treated the same way without diesel exhaust. The effects within lung and other tissues were assessed by immunohistochemistry of formalin-fixed and paraffin-embedded tissues. Levels of inflammation and citrullination related markers were investigated. Levels of parenchymal damage were also measured. Results: In vivo study corroborates our own data from in vitro and reveals diesel exhaust initiated inflammatory shift and modulation of lung peptidyl arginine deiminase 4 (PAD4), citrullination associated enzyme, levels. In addition, high levels of citrulline were observed in exposed lung tissue sections co-localising with increased parenchymal destruction. Conclusions: Subacute exposure to diesel exhaust renders mice lungs inflammatory and modifies certain structural proteins. Such structural changes of proteins may pave a pathways to lost/gain function of affected molecules and also propagate autoimmune processes within the lung and systemically.

Keywords: air pollution, citrullination, in vivo, lungs

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Authors: Sree Sanyal

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Research in ageing often highlights the age-related health dimension more than the psycho-social characteristics of the elderly, which also influences and challenges the health outcomes. Multimorbidity is defined as the person having more than one chronic non-communicable diseases and their prevalence increases with ageing. The study aims to evaluate the influence of social ties on self-reported prevalence of multimorbidity (selected chronic non-communicable diseases) among the selected states of elderly population in India. The data is accessed from Building Knowledge Base on Population Ageing in India (BKPAI), collected in 2011 covering the self-reported chronic non-communicable diseases like arthritis, heart disease, diabetes, lung disease with asthma, hypertension, cataract, depression, dementia, Alzheimer’s disease, and cancer. The data of the above diseases were taken together and categorized as: ‘no disease’, ‘one disease’ and ‘multimorbidity’. The predicted variables were demographic, socio-economic, residential types, and the variable of social ties includes social support, social engagement, perceived support, connectedness, and importance of the elderly. Predicted probability for multiple logistic regression was used to determine the background characteristics of the old in association with chronic morbidities showing multimorbidity. The finding suggests that 24.35% of the elderly are suffering from multimorbidity. Research shows that with reference to ‘no disease’, according to the socio-economic characteristics of the old, the female oldest old (80+) from others in caste and religion, widowed, never had any formal education, ever worked in their life, coming from the second wealth quintile standard, from rural Maharashtra are more prone with ‘one disease’. From the social ties background, the elderly who perceives they are important to the family, after getting older their decision-making status has been changed, prefer to stay with son and spouse only, satisfied with the communication from their children are more likely to have less single morbidity and the results are significant. Again, with respect to ‘no disease’, the female oldest old (80+), who are others in caste, Christian in religion, widowed, having less than 5 years of education completed, ever worked, from highest wealth quintile, residing in urban Kerala are more associated with multimorbidity. The elderly population who are more socially connected through family visits, public gatherings, gets support in decision making, who prefers to spend their later years with son and spouse only but stays alone shows lesser prevalence of multimorbidity. In conclusion, received and perceived social integration and support from associated neighborhood in the older days, knowing about their own needs in life facilitates better health and wellbeing of the elderly population in selected states of India.

Keywords: morbidity, multi-morbidity, prevalence, social ties

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Authors: Arash Dalili, Hamed Tahmouressi, Mina Hoorfar

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Advances in lab-on-a-chip (LOC) devices have led to significant advances in the manipulation, separation, and isolation of particles and cells. Among the different active and passive particle manipulation methods, dielectrophoresis (DEP) has been proven to be a versatile mechanism as it is label-free, cost-effective, simple to operate, and has high manipulation efficiency. DEP has been applied for a wide range of biological and environmental applications. A popular form of DEP devices is the continuous manipulation of particles by using co-planar slanted electrodes, which utilizes a sheath flow to focus the particles into one side of the microchannel. When particles enter the DEP manipulation zone, the negative DEP (nDEP) force generated by the slanted electrodes deflects the particles laterally towards the opposite side of the microchannel. The lateral displacement of the particles is dependent on multiple parameters including the geometry of the electrodes, the width, length and height of the microchannel, the size of the particles and the throughput. In this study, COMSOL Multiphysics® modeling along with experimental studies are used to investigate the effect of the aforementioned parameters. The electric field between the electrodes and the induced DEP force on the particles are modelled by COMSOL Multiphysics®. The simulation model is used to show the effect of the DEP force on the particles, and how the geometry of the electrodes (width of the electrodes and the gap between them) plays a role in the manipulation of polystyrene microparticles. The simulation results show that increasing the electrode width to a certain limit, which depends on the height of the channel, increases the induced DEP force. Also, decreasing the gap between the electrodes leads to a stronger DEP force. Based on these results, criteria for the fabrication of the electrodes were found, and soft lithography was used to fabricate interdigitated slanted electrodes and microchannels. Experimental studies were run to find the effect of the flow rate, geometrical parameters of the microchannel such as length, width, and height as well as the electrodes’ angle on the displacement of 5 um, 10 um and 15 um polystyrene particles. An empirical equation is developed to predict the displacement of the particles under different conditions. It is shown that the displacement of the particles is more for longer and lower height channels, lower flow rates, and bigger particles. On the other hand, the effect of the angle of the electrodes on the displacement of the particles was negligible. Based on the results, we have developed an optimum design (in terms of efficiency and throughput) for three size separation of particles.

Keywords: COMSOL Multiphysics, Dielectrophoresis, Microfluidics, Particle separation

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Authors: Debananda Mohapatra, Subramanya Badrayyana, Smrutiranjan Parida

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Carbon nano-onions (CNOs) are the spherical graphitic nanostructures composed of concentric shells of graphitic carbon can be hypothesized as the intermediate state between fullerenes and graphite. These are very important members in fullerene family also known as the multi-shelled fullerenes can be envisioned as promising supercapacitor electrode with high energy & power density as they provide easy access to ions at electrode-electrolyte interface due to their curvature. There is still very sparse report concerning on CNOs as electrode despite having an excellent electrodechemical performance record due to their unavailability and lack of convenient methods for their high yield preparation and purification. Keeping all these current pressing issues in mind, we present a facile scalable and straightforward flame synthesis method of pure and highly dispersible CNOs without contaminated by any other forms of carbon; hence, a post processing purification procedure is not necessary. To the best of our knowledge, this is the very first time; we developed an extremely simple, light weight, novel inexpensive, flexible free standing pristine CNOs electrode without using any binder element. Locally available daily used cotton wipe has been used for fabrication of such an ideal electrode by ‘dipping and drying’ process providing outstanding stretchability and mechanical flexibility with strong adhesion between CNOs and porous wipe. The specific capacitance 102 F/g, energy density 3.5 Wh/kg and power density 1224 W/kg at 20 mV/s scan rate are the highest values that ever recorded and reported so far in symmetrical two electrode cell configuration with 1M Na2SO4 electrolyte; indicating a very good synthesis conditions employed with optimum pore size in agreement with electrolyte ion size. This free standing CNOs electrode also showed an excellent cyclic performance and stability retaining 95% original capacity after 5000 charge –discharge cycles. Furthermore, this unique method not only affords binder free - freestanding electrode but also provide a general way of fabricating such multifunctional promising CNOs based nanocomposites for their potential device applications in flexible solar cells and lithium-ion batteries.

Keywords: binder-free, flame synthesis, flexible, carbon nano onion

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Authors: Babarina Maria, Andropova Margarita

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Introduction: Hypoparathyroidism (HypoPT) is a rare endocrine disorder with an estimated prevalence of 0.25 per 1000 individuals. The most common cause of HypoPT is the loss of active parathyroid tissue following thyroid or parathyroid surgery. Sometimes permanent postoperative HypoPT occures, manifested by hypocalcemia in combination with low levels of PTH during 6 months or more after surgery. Cognitive impairments in patients with hypocalcemia due to chronic HypoPT are observed, and this can lead to problems and challenges in everyday living: memory loss and impaired concentration, that may be the cause of poor compliance. Clinical case: Patient K., 66 years old, underwent thyroidectomy in 2013 (at the age of 55) because of papillary thyroid cancer T1NxMx, histopathology findings confirmed the diagnosis. 5 years after the surgery, she was followed up on an outpatient basis, TSH levelsonly were monitored, and the dose of levothyroxine was adjusted. In 2018 due to, increasing complaints include tingling and cramps in the arms and legs, memory loss, sleep disorder, fatigue, anxiety, hair loss, muscle pain, tachycardia, positive Chvostek, and Trousseau signs were diagnosed during examination, also in blood analyses: total Ca 1.86 mmol/l (2.15-2.55), Ca++ 0.96 mmol/l (1.12-1.3), P 1.55 mmol/l (0.74-1.52), Mg 0.79 mmol/l (0.66-1.07) - chronic postoperative HypoPT was diagnosed. Therapy was initiated: alfacalcidol 0.5 mcg per day, calcium carbonate 2000 mg per day, cholecalciferol 1000 IU per day, magnesium orotate 3000 mg per day. During the case follow-up, hypocalcemia, hyperphosphatemia persisted, hypercalciuria15.7 mmol/day (2.5-6.5) was diagnosed. Dietary recommendations were given because of the high content of phosphorus rich foods, and therapy was adjusted: the dose of alfacalcidol was increased to 2.5 mcg per day, and the dose of calcium carbonate was reduced to 1500 mg per day. As part of the screening for complications of hypoPT, data for cataracts, Fahr syndrome, nephrocalcinosis, and kidney stone disease were not obtained. However, HypoPT compensation was not achieved, and therefore hydrochlorothiazide 25 mg was initiated, the dose of alfacalcidol was increased to 3 mcg per day, calcium carbonate to 3000 mg per day, magnesium orotate and cholecalciferol were continued at the same doses. Therapeutic goals were achieved: calcium phosphate product <4.4 mmol2/l2, there were no episodes of hypercalcemia, twenty-four-hour urinary calcium excretion was significantly reduced. Conclusion: Timely prescription, careful explanation of drugs usage rules, and monitoring and maintaining blood and urine parameters within the target contribute to the prevention of HypoPT complications development and life-threatening events.

Keywords: hypoparathyroidism, hypocalcemia, hyperphosphatemia, hypercalciuria

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Authors: Irati Barandiaran, Xabier Velasco-Iza, Galder Kortaberria

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Hybrid inorganic/organic nanostructured materials based on block copolymers are of considerable interest in the field of Nanotechnology, taking into account that these nanocomposites combine the properties of polymer matrix and the unique properties of the added nanoparticles. The use of block copolymers as templates offers the opportunity to control the size and the distribution of inorganic nanoparticles. This research is focused on the surface modification of inorganic nanoparticles to reach a good interface between nanoparticles and polymer matrices which hinders the nanoparticle aggregation. The aim of this work is to obtain a good and selective dispersion of Fe3O4 magnetic nanoparticles into different types of block copolymers such us, poly(styrene-b-methyl methacrylate) (PS-b-PMMA), poly(styrene-b-ε-caprolactone) (PS-b-PCL) poly(isoprene-b-methyl methacrylate) (PI-b-PMMA) or poly(styrene-b-butadiene-b-methyl methacrylate) (SBM) by using different grafting strategies. Fe3O4 magnetic nanoparticles have been surface-modified with polymer or block copolymer brushes following different grafting methods (grafting to, grafting from and grafting through) to achieve a selective location of nanoparticles into desired domains of the block copolymers. Morphology of fabricated hybrid nanocomposites was studied by means of atomic force microscopy (AFM) and with the aim to reach well-ordered nanostructured composites different annealing methods were used. Additionally, nanoparticle amount has been also varied in order to investigate the effect of the nanoparticle content in the morphology of the block copolymer. Nowadays different characterization methods were using in order to investigate magnetic properties of nanometer-scale electronic devices. Particularly, two different techniques have been used with the aim of characterizing synthesized nanocomposites. First, magnetic force microscopy (MFM) was used to investigate qualitatively the magnetic properties taking into account that this technique allows distinguishing magnetic domains on the sample surface. On the other hand, magnetic characterization by vibrating sample magnetometer and superconducting quantum interference device. This technique demonstrated that magnetic properties of nanoparticles have been transferred to the nanocomposites, exhibiting superparamagnetic behavior similar to that of the maghemite nanoparticles at room temperature. Obtained advanced nanostructured materials could found possible applications in the field of dye-sensitized solar cells and electronic nanodevices.

Keywords: atomic force microscopy, block copolymers, grafting techniques, iron oxide nanoparticles

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Authors: Ahlam Ali, Katrina Lappin, Jaine Blayney, Ken Mills

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Leukaemia is the most frequently (30%) occurring type of paediatric cancer. Of these, approximately 80% are acute lymphoblastic leukaemia (ALL) with acute myeloid leukaemia (AML) cases making up the remaining 20% alongside other leukaemias. Unfortunately, children with AML do not have promising prognosis with only 60% surviving 5 years or longer. It has been highlighted recently the need for age-specific therapies for AML patients, with paediatric AML cases having a different mutational landscape compared with AML diagnosed in adult patients. Drug Repurposing is a recognized strategy in drug discovery and development where an already approved drug is used for diseases other than originally indicated. We aim to identify novel combination therapies with the promise of providing alternative more effective and less toxic induction therapy options. Our in-silico analysis highlighted ‘cell death and survival’ as an aberrant, potentially targetable pathway in paediatric AML patients. On this basis, 83 apoptotic inducing compounds were screened. A preliminary single agent screen was also performed to eliminate potentially toxic chemicals, then drugs were constructed into a pooled library with 10 drugs per well over 160 wells, with 45 possible pairs and 120 triples in each well. Seven cell lines were used during this study to represent the clonality of AML in paediatric patients (Kasumi-1, CMK, CMS, MV11-14, PL21, THP1, MOLM-13). Cytotoxicity was assessed up to 72 hours using CellTox™ Green reagent. Fluorescence readings were normalized to a DMSO control. Z-Score was assigned to each well based on the mean and standard deviation of all the data. Combinations with a Z-Score <2 were eliminated and the remaining wells were taken forward for further analysis. A well was considered ‘successful’ if each drug individually demonstrated a Z-Score <2, while the combination exhibited a Z-Score >2. Each of the ten compounds in one well (155) had minimal or no effect as single agents on cell viability however, a combination of two or more of the compounds resulted in a substantial increase in cell death, therefore the ten compounds were de-convoluted to identify a possible synergistic pair/triple combinations. The screen identified two possible ‘novel’ drug pairing, with BCL2 inhibitor ABT-737, combined with either a CDK inhibitor Purvalanol A, or AKT/ PI3K inhibitor LY294002. (ABT-737- 100 nM+ Purvalanol A- 1 µM) (ABT-737- 100 nM+ LY294002- 2 µM). Three possible triple combinations were identified (LY2409881+Akti-1/2+Purvalanol A, SU9516+Akti-1/2+Purvalanol A, and ABT-737+LY2409881+Purvalanol A), which will be taken forward for examining their efficacy at varying concentrations and dosing schedules, across multiple paediatric AML cell lines for optimisation of maximum synergy. We believe that our combination screening approach has potential for future use with a larger cohort of drugs including FDA approved compounds and patient material.

Keywords: AML, drug repurposing, ABT-737, apoptosis

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Authors: Ibtisam A. Abbas Al-Darkazly

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In this study, the mechanical properties of alginate hydrogel material for self-standing 3D scaffold architecture with proper shape fidelity are investigated. In-lab built 3D bio-printer extrusion-based technology is utilized to fabricate 3D alginate scaffold constructs. The pressure, needle speed and stage speed are varied using a computer-controlled system. The experimental result indicates that the concentration of alginate solution, calcium chloride (CaCl₂) cross-linking concentration and cross-linking ratios lead to the formation of alginate hydrogel with various gelation states. Besides, the gelling conditions, such as cross-linking reaction time and temperature also have a significant effect on the mechanical properties of alginate hydrogel. Various experimental tests such as the material gelation, the material spreading and the printability test for filament collapse as well as the swelling test were conducted to evaluate the fabricated 3D scaffold constructs. The result indicates that the fabricated 3D scaffold from composition of 3.5% wt alginate solution, that is prepared in DI water and 1% wt CaCl₂ solution with cross-linking ratios of 7:3 show good printability and sustain good shape fidelity for more than 20 days, compared to alginate hydrogel that is prepared in a phosphate buffered saline (PBS). The fabricated self-standing 3D scaffold constructs measured 30 mm × 30 mm and consisted of 4 layers (n = 4) show good pore geometry and clear grid structure after printing. In addition, the percentage change of swelling degree exhibits high swelling capability with respect to time. The swelling test shows that the geometry of 3D alginate-scaffold construct and of the macro-pore are rarely changed, which indicates the capability of holding the shape fidelity during the incubation period. This study demonstrated that the mechanical and physical properties of alginate hydrogel could be tuned for a 3D bio-printing extrusion-based system to fabricate self-standing 3D scaffold soft structures. This 3D bioengineered scaffold provides a natural microenvironment present in the extracellular matrix of the tissue, which could be seeded with the biological cells to generate the desired 3D live tissue model for in vitro and in vivo tissue engineering applications.

Keywords: biomaterial, calcium chloride, 3D bio-printing, extrusion, scaffold, sodium alginate, tissue engineering

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Authors: Basil Nkosilathi Dube, Wilson R. Nyemba, Panashe Mandevu

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In highway driving, over 50 percent of the power produced by the engine is used to overcome aerodynamic drag, which is a force that opposes a body’s motion through the air. Aerodynamic drag and thus fuel consumption increase rapidly at speeds above 90kph. It is desirable to minimize fuel consumption. Aerodynamic drag reduction in highway driving is the best approach to minimize fuel consumption and to reduce the negative impacts of greenhouse gas emissions on the natural environment. Fuel economy is the ultimate concern of automotive development. This study aims to design and analyze drag-reducing devices for a Mazda T3500 truck, namely, the cab roof and rear (trailer tail) fairings. The aerodynamic effects of adding these append devices were subsequently investigated. To accomplish this, two 3D CAD models of the Mazda truck were designed using the Design Modeler. One, with these, append devices and the other without. The models were exported to ANSYS Fluent for computational fluid dynamics analysis, no wind tunnel tests were performed. A fine mesh with more than 10 million cells was applied in the discretization of the models. The realizable k-ε turbulence model with enhanced wall treatment was used to solve the Reynold’s Averaged Navier-Stokes (RANS) equation. In order to simulate the highway driving conditions, the tests were simulated with a speed of 100 km/h. The effects of these devices were also investigated for low-speed driving. The drag coefficients for both models were obtained from the numerical calculations. By adding the cab roof and rear (trailer tail) fairings, the simulations show a significant reduction in aerodynamic drag at a higher speed. The results show that the greatest drag reduction is obtained when both devices are used. Visuals from post-processing show that the rear fairing minimized the low-pressure region at the rear of the trailer when moving at highway speed. The rear fairing achieved this by streamlining the turbulent airflow, thereby delaying airflow separation. For lower speeds, there were no significant differences in drag coefficients for both models (original and modified). The results show that these devices can be adopted for improving the aerodynamic efficiency of the Mazda T3500 truck at highway speeds.

Keywords: aerodynamic drag, computation fluid dynamics, fluent, fuel consumption

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Authors: Alexandra Emmanuelle Membangbi, Jacky Njiki Bikoï, Esther Del-florence Moni Ndedi, Marie Joseph Nkodo Mindimi, Donatien Serge Mbaga, Elsa Nguiffo Makue, André Chris Mikangue Mbongue, Martha Mesembe, George Ikomey Mondinde, Eric Walter Perfura-yone, Sara Honorine Riwom Essama

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Background: Tuberculosis (TB) is one of the top lethal infectious diseases worldwide. In recent years, interferon-γ (INF-γ) release assays (IGRAs) have been established as routine tests for diagnosing TB infection. However, produced INF-γ assessment failed to distinguish active TB (ATB) from latent TB infection (LTBI), especially in TB epidemic areas. In addition to IFN-γ, interleukin-2 (IL-2), another cytokine secreted by activated T cells, is also involved in immune response against Mycobacterium tuberculosis. The aim of the study was to assess the capacity of IFN-γ and IL2 to evaluate the therapeutic response of patients on anti-tuberculosis treatment. Material and Methods: We conducted a cross-sectional study in the Pneumonology Departments of the Jamot Hospital in Yaoundé between May and August 2021. After signed the informed consent, the sociodemographic data, as well as 5 mL of blood, were collected in the crook of the elbow of each participant. Sixty-one subjects were selected (n= 61) and divided into 4 groups as followed: group 1: resistant tuberculosis (n=13), group 2: active tuberculosis (n=19), group 3 cured tuberculosis (n=16), and group 4: presumed healthy persons (n=13). The cytokines of interest were determined using an indirect Enzyme-linked Immuno-Sorbent Assay (ELISA) according to the manufacturer's recommendations. P-values < 0.05 were interpreted as statistically significant. All statistical calculations were performed using SPSS version 22.0 Results: The results showed that men were more 14/61 infected (31,8%) with a high presence in active and resistant TB groups. The mean age was 41.3±13.1 years with a 95% CI = [38.2-44.7], the age group with the highest infection rate was ranged between 31 and 40 years. The IL-2 and INF-γ means were respectively 327.6±160.6 pg/mL and 26.6±13.0 pg/mL in active tuberculosis patients, 251.1±30.9 pg/mL and 21.4±9.2 pg/mL in patients with resistant tuberculosis, while it was 149.3±93.3 pg/mL and 17.9±9.4 pg/mL in cured patients, 15.1±8.4 pg/mL and 5.3±2.6 pg/mL in participants presumed healthy (p <0.0001). Significant differences in IFN-γ and IL-2 rates were observed between the different groups. Conclusion: Monitoring the serum levels of INF-γ and IL-2 would be useful to evaluate the therapeutic response of anti-tuberculosis patients, particularly in the both cytokines association case, that could improve the accuracy of routine examinations.

Keywords: antibiotic therapy, interferon gamma, interleukin 2, tuberculosis

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Authors: Christoph Hacker, Zeynep Karahaliloglu, Gunnar Seide, Emir Baki Denkbas, Thomas Gries

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A wound dressing material is designed to facilitate wound healing and minimize scarring. An ideal wound dressing material should protect the wound from any contaminations of exogeneous microorganism. In addition, the dressing material should provide a moist environment through extraction of body fluid from the wound area. Recently, wound dressing electrospun nanofibrous membranes are produced by electrospinning from a polymer solution or a polymer melt. These materials have a great potential as dressing materials for wound healing because of superior properties such as high surface-to-volume ratio, high porosity with excellent pore interconnectivity. Melt electrospinning is an attractive tissue engineering scaffold manufacturing process which eliminated the health risk posed by organic solvents used in electrospinning process and reduced the production costs. In this study, antibacterial wound dressing materials were prepared from TPU (Elastollan 1185A) by a melt-electrospinning technique. The electrospinning parameters for an efficient melt-electrospinning process of TPU were optimized. The surface of the fibers was modified with poly(ethylene glycol) (PEG) by radio-frequency glow discharge plasma deposition method and with silver nanoparticles (nAg) to improve their wettability and antimicrobial properties. TPU melt-electrospun mats were characterized using SEM, DSC, TGA and XPS. The cell viability and proliferation on modified melt-electrospun TPU mats were evaluated using a mouse fibroblast cell line (L929). Antibacterial effects of theirs against both Staphylococcus aureus strain and Escherichia coli were investigated by disk-diffusion method. TPU was successfully processed into a porous, fibrous network of beadless fibers in the micrometer range (4.896±0.94 µm) with a voltage of 50 kV, a working distance of 6 cm, a temperature of the thermocouple and hot coil of 225–230ºC, and a flow rate of 0.1 mL/h. The antibacterial test indicated that PEG-modified nAg-loaded TPU melt-electrospun structure had excellent antibacterial effects and cell study results demonstrated that nAg-loaded TPU mats had no cytotoxic effect on the fibroblast cells. In this work, the surface of a melt-electrospun TPU mats was modified via PEG monomer and then nAg. Results showed melt-electrospun TPU mats modified with PEG and nAg have a great potential for use as an antibacterial wound dressing material and thus, requires further investigation.

Keywords: melt electrospinning, nanofiber, silver nanoparticles, wound dressing

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Authors: Sharat Chandra, Zilong Wang, Ru-Rong Ji, Andrey Bortsov

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Chronic pain (CP) therapeutic approaches have limited efficacy. As a result, doctors are prescribing opioids for chronic pain, leading to opioid overuse, abuse, and addiction epidemic. Therefore, the development of effective and safe CP drugs remains an unmet medical need. Voltage-gated sodium (Nav) channels act as cardiovascular and neurological disorder’s molecular targets. Nav channels selective inhibitors are hard to design because there are nine closely-related isoforms (Nav1.1-1.9) that share the protein sequence segments. We are targeting the Nav1.7 found in the peripheral nervous system and engaged in the perception of pain. The objective of this project was to screen a 1.5 million compound library for identification of inhibitors for Nav1.7 with analgesic effect. In this study, we designed a protocol for identification of isoform-selective inhibitors of Nav1.7, by utilizing the prior information on isoform-selective antagonists. First, a similarity search was performed; then the identified hits were docked into a binding site on the fourth voltage-sensor domain (VSD4) of Nav1.7. We used the FTrees tool for similarity searching and library generation; the generated library was docked in the VSD4 domain binding site using FlexX and compounds were shortlisted using a FlexX score and SeeSAR hyde scoring. Finally, the top 25 compounds were tested with molecular dynamics simulation (MDS). We reduced our list to 9 compounds based on the MDS root mean square deviation plot and obtained them from a vendor for in vitro and in vivo validation. Whole-cell patch-clamp recordings in HEK-293 cells and dorsal root ganglion neurons were conducted. We used patch pipettes to record transient Na⁺ currents. One of the compounds reduced the peak sodium currents in Nav1.7-HEK-293 stable cell line in a dose-dependent manner, with IC50 values at 0.74 µM. In summary, our computer-aided analgesic discovery approach allowed us to develop pre-clinical analgesic candidate with significant reduction of time and cost.

Keywords: chronic pain, voltage-gated sodium channel, isoform-selective antagonist, similarity search, virtual screening, analgesics development

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Authors: Norazlan Mohmad Misnan, Maizatul Hasyima Omar, Mohd Isa Wasiman

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Papaya (Carica papaya L., Caricaceae) is a tree which mainly cultivated for its fruits in many tropical regions including Australia, Brazil, China, Hawaii, and Malaysia. Beside of fruits, its leaves, seeds, and latex have also been traditionally used for treating diseases, which also reported to possess anti-cancer and anti- malaria properties. Its leaves have been reported to consist of various chemical compounds such as alkaloids, flavonoids and phenolics. Clitorin and manghaslin are among major flavonoids presence. Thus, the aim of this study is to quantify the purity of these isolated compounds (clitorin and manghsalin) by using quantitative Nuclear Magnetic Resonance (qNMR) analysis. Only fresh C. papaya leaves were used for juice extraction procedure and subsequently was freeze-dried to obtain a dark green powdered form of the extract prior to Centrifugal Partition Chromatography (CPC) separation. The CPC experiments were performed using a two-phase solvent system comprising ethyl acetate/butanol/water (1:4:5, v/v/v/v) solvent. The upper organic phase was used as the stationary phase, and the lower aqueous phase was employed as the mobile phase. Ten fractions were obtained after an hour runtime analysis. Fraction 6 and fraction 8 has been identified as clitorin (m/z 739.21 [M-H]-) and manghaslin (m/z 755.21 [M-H]-), respectively, based on LCMS data and full analysis of NMR (1H NMR, 13C NMR, HMBC, and HSQC). The 1H-qNMR measurements were carried out using a 400 MHz NMR spectrometer (JEOL ECS 400MHz, Japan) and deuterated methanol was used as a solvent. Quantification was performed using the AQARI method (Accurate Quantitative NMR) with deuterated 1,4-Bis(trimethylsilyl)benzene (BTMSB) as an internal reference substances. This AQARI protocol includes not only NMR measurement but also sample preparation that provide highest precision and accuracy than other qNMR methods. The 90° pulse length and the T1 relaxation times for compounds and BTMSB were determined prior to the quantification to give the best signal-to-noise ratio. Regions containing the two downfield signals from aromatic part (6.00–6.89 ppm), and the singlet signal, (18H) arising from BTMSB (0.63-1.05ppm) were selected for integration. The purity of clitorin and manghaslin were calculated to be 52.22% and 43.36%, respectively. Further purification is needed in order to increase its purity. This finding has demonstrated the use of qNMR for quality control and standardization of various plant extracts and which can be applied for NMR fingerprinting of other plant-based products with good reproducibility and in the case where commercial standards is not readily available.

Keywords: Carica papaya, clitorin, manghaslin, quantitative Nuclear Magnetic Resonance, Centrifugal Partition Chromatography

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Authors: Amanda Sheard, Garry Lee, Katherine Stockham

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Introduction: Health benefits associated with wine consumption have been well documented; these include anticancer, anti-inflammatory, and cardiovascular protection. The majority of these health benefits have been linked to polyphenols found within wine and grapes. Once consumed polyphenols exhibit free radical quenching capabilities. Environmental factors such as rainfall, temperature, CO2 levels and sunlight exposure have been shown to affect the polyphenol content of grapes. The objective of this work was to evaluate the effect of growing conditions on the antioxidant capacity of grapes obtained from a single plot vineyard in Perth. This was achieved through the analysis of samples using; oxygen radical antioxidant capacity (ORAC), cellular antioxidant activity (CAA) in human red blood cells, ICP-MS and ICP-OES, total polyphenols (PP’s), and total flavonoid’s (FLa). The data obtained was compared to observed climate data. The 14 Selected Vitis Vinefera L. cultivars included Cabernet franc, Cabernet Sauvignon, Carnelian, Chardonnay, Grenache, Melbec, Merlot, Orange muscat, Rousanne, Sauvignon Blanc, Shiraz, Tempernillo, Verdelho, and Voignier. Results: Notable variation’s between cultivars included results ranging from 125 mg/100 g-350 mg/100 g for PP’s, 93 mg/100 g–300 mg/100 g for FLa, 13 mM T.E/kg–33 mM T.E/kg for ORAC and 0.3 mM Q.E/kg–27 mM Q.E/kg CAA were found between red and white grape cultivars. No correlation was found between CAA and the ORAC obtained in this study; except that white cultivars were consistently lower than red. ICP analysis showed that seeds contained the highest concentration of copper followed by skins and flesh of the grape. A positive correlation between copper and ORAC was found. The ORAC, PP’s, and FLa in red grapes were consistently higher than white grape cultivars; these findings were supported by literature values. Significance: The cellular antioxidant activities of white and red wine cultivars were used to compare the bioactivity of these grapes against the chemical ORAC measurement. The common method of antioxidant activity measurement is the chemical value from ORAC analysis; however this may not reflect the activity within the human body. Hence, the measurements were also carried out using the cellular antioxidant activity to perform a comparison. Additionally, the study explored the influence of weather systems such as El Niño and La Niña on the polyphenol content of Australian wine cultivars grown in Perth.

Keywords: oxygen radical antioxidant activity, cellular antioxidant activity, total polyphenols, total flavonoids, wine grapes, climate

Procedia PDF Downloads 287

Authors: Hamed Ahari, Behrouz Akbari Adreghani, Vadood Razavilar, Amirali Anvar, Sima Moradi, Hourieh Shalchi

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Considering the ever increasing population and industrialization of the developmental trend of humankind's life, we are no longer able to detect the toxins produced in food products using the traditional techniques. This is due to the fact that the isolation time for food products is not cost-effective and even in most of the cases, the precision in the practical techniques like the bacterial cultivation and other techniques suffer from operator errors or the errors of the mixtures used. Hence with the advent of nanotechnology, the design of selective and smart sensors is one of the greatest industrial revelations of the quality control of food products that in few minutes time, and with a very high precision can identify the volume and toxicity of the bacteria. Methods and Materials: In this technique, based on the bacterial antibody connection to nanoparticle, a sensor was used. In this part of the research, as the basis for absorption for the recognition of bacterial toxin, medium sized silica nanoparticles of 10 nanometer in form of solid powder were utilized with Notrino brand. Then the suspension produced from agent-linked nanosilica which was connected to bacterial antibody was positioned near the samples of distilled water, which were contaminated with Staphylococcus aureus bacterial toxin with the density of 10-3, so that in case any toxin exists in the sample, a connection between toxin antigen and antibody would be formed. Finally, the light absorption related to the connection of antigen to the particle attached antibody was measured using spectrophotometry. The gene of 23S rRNA that is conserved in all Staphylococcus spp., also used as control. The accuracy of the test was monitored by using serial dilution (l0-6) of overnight cell culture of Staphylococcus spp., bacteria (OD600: 0.02 = 107 cell). It showed that the sensitivity of PCR is 10 bacteria per ml of cells within few hours. Result: The results indicate that the sensor detects up to 10-4 density. Additionally, the sensitivity of the sensors was examined after 60 days, the sensor by the 56 days had confirmatory results and started to decrease after those time periods. Conclusions: Comparing practical nano biosensory to conventional methods like that culture and biotechnology methods(such as polymerase chain reaction) is accuracy, sensitiveness and being unique. In the other way, they reduce the time from the hours to the 30 minutes.

Keywords: exotoxin, nanobiosensor, recognition, Staphylococcus aureus

Procedia PDF Downloads 385

Authors: Mirijam Vrabec, Marian Janak, Bojan Ambrozic, Angelja K. Surca, Nastja Rogan Smuc, Nina Zupancic, Saso Sturm

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Natural microdiamonds and moissanite (SiC) can form during the orogenic events under ultrahigh-pressure metamorphic conditions (UHP), when parts of Earth’s crust are subducted to extreme depths. So far, such processes were identified only in few places on the Earth, and therefore, represent unique opportunity to study the evolution of the Earth’s deep interior. An important discovery of microdiamonds and moissanite was reported from Pohorje, (Slovenia), where they occurred as single or polyphase inclusions in garnets. Metasedimentary rocks from Pohorje are predominantly gneisses representing parts of the Austroalpine metamorphic units of the Eastern Alps. During Cretaceous orogeny, (ca. 95–92 Ma) continental crustal rocks were deeply subducted to the mantle depths (below 100 km) and metamorphosed at pressures exceeding 3.5 GPa and temperatures between 800–850 °C. Microstructural and phase analysis of the inclusions as well as detailed elemental analysis of host garnets were carried out combining several analytical techniques: optical microscope in plane polarized transmitted light, electron probe microanalysis (EPMA) with wavelength-dispersive x-ray spectrometry (WDS) and field-emission scanning microscope (FEG-SEM) with energy-dispersive x-ray spectroscopy (EDS). Micro-Raman analysis revealed sharp, first order diamond bands sometimes accompanied by graphite bands implying that transformation of diamond back to graphite occurred. To study the chemical and crystallographic relationship between microdiamonds and co-inclusions, advanced techniques of transmission electron microscopy (TEM) were applied, which included high-angle annular dark-field scanning transmission electron microscopy (HAADF-STEM), combined with EDS and electron energy-loss spectroscopy (EELS). To prepare electron transparent TEM lamellae selectively a dual-beam Focused Ion Beam/SEM (FIB/SEM) was employed. Detailed study of TEM lamellae, which was cross-sectioned from the highly faceted inclusion body located within the host garnet crystal matrix, revealed rich and rather complex internal structure. Namely, the negative crystal facets of the main inclusion body were typically decorated with up to 1 μm thick amorphous layer, reflecting the general garnet composition with slight variations in Fe/Ca content. Within these layers, ELNES analysis revealed the presence of a 28–30 nm thick layer of amorphous carbon. The very last section of this layer corresponds to composition of SiO2. Within the inclusion, besides diamond and moissanite alumosilicate mineral with pronounced layered structure, iron sulfides and chlorine were identified under TEM and CO2 and CH4 using Raman. Moissanite is found as single crystal or composed from numerous highly textured nano-crystals with the average size of 10 nm. Moissanite inclusions were found embedded inside the amorphous crust implying that moissanite crystalized well before the deposition of the amorphous layer. From the microstructural, crystallographic and chemical observations so far we can deduce, that polyphase inclusions in diamond bearing garnets from Pohorje most probably crystallized from reduced supercritical fluids. Based on layered interface structure of the host mineral multiphase process of crystallization is possible. The presence of microdiamonds and moissanite in rocks from Pohorje demonstrates that these parts of the Eastern Alps were subducted to extreme depths, and were subsequently exhumed back to the Earth's surface without complete breakdown of UHP mineral phases, allowing a rear and exceptional opportunity to study them in-situ.

Keywords: diamond, fluid inclusions, moissanite, TEM, UHP metamorphism.

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Authors: S. Rodosik, J. P. Poirot-Crouvezier, Y. Bultel

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With the expansion of the hydrogen economy, Proton Exchange Membrane Fuel Cell (PEMFC) systems are often presented as promising energy converters suitable for transport applications. However, reaching a durability of 5000 h recommended by the U.S. Department of Energy and decreasing system cost are still major hurdles to their development. In order to increase the system efficiency and simplify the system without affecting the fuel cell lifetime, an architecture called alternative fuel feeding has been developed. It consists in a fuel cell stack divided into two parts, alternatively fed, implemented on a 5-kW system for real scale testing. The operation strategy can be considered close to Dead End Anode (DEA) with specific modifications to avoid water and nitrogen accumulation in the cells. The two half-stacks are connected in series to enable each stack to be alternatively fed. Water and nitrogen accumulated can be shifted from one half-stack to the other one according to the alternative feeding frequency. Thanks to the homogenization of water vapor along the stack, water management was improved. The operating conditions obtained at system scale are close to recirculation without the need of a pump or an ejector. In a first part, a performance comparison with the DEA strategy has been performed. At high temperature and low pressure (80°C, 1.2 bar), performance of alternative fuel feeding was higher, and the system efficiency increased. In a second part, in order to highlight the benefits of the architecture on the fuel cell lifetime, two durability tests, lasting up to 1000h, have been conducted. A test on the 5-kW system has been compared to a reference test performed on a test bench with a shorter stack, conducted with well-controlled operating parameters and flow-through hydrogen strategy. The durability test is based upon the Fuel Cell Dynamic Load Cycle (FC-DLC) protocol but adapted to the system limitations: without OCV steps and a maximum current density of 0.4 A/cm². In situ local measurements with a segmented S++® plate performed all along the tests, showed a more homogeneous distribution of the current density with alternative fuel feeding than in flow-through strategy. Tests performed in this work enabled the understanding of this architecture advantages and drawbacks. Alternative fuel feeding architecture appeared to be a promising solution to ensure the humidification function at the anode side with a simplified fuel cell system.

Keywords: automotive conditions, durability, fuel cell system, proton exchange membrane fuel cell, stack architecture

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Authors: Haris Setiawan, Phongsakorn Chuammitri, Korawan Sringarm, Montira Intanon, Anucha Sathanawongs

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Capacitation is a prerequisite to achieving sperm competency to penetrate the oocyte naturally occurring in vivo throughout the female reproductive tract and entangling secretory fluid and epithelial cells. One of the crucial compounds in the oviductal fluid which promotes capacitation is albumin, secreted in major concentrations. However, the difficulties in the collection and the inconsistency of the oviductal fluid composition throughout the estrous cycle have replaced its function with serum-based albumins such as bovine serum albumin (BSA). BSA has been primarily involved and evidenced for their stabilizing effect to maintain the acrosome intact during the capacitation process, modulate hyperactivation, and elevate the number of sperm bound to zona pellucida. Contrary to its benefits, the use of blood-derived products in the culture system is not sustainable and increases the risk of disease transmissions, such as Creutzfeldt-Jakob disease (CJD) and bovine spongiform encephalopathy (BSE). Moreover, it has been asserted that this substance is an aeroallergen that produces allergies and respiratory problems. In an effort to identify an alternative sustainable and non-toxic albumin source, the present work evaluated sperm reactions to a capacitation medium containing albumin derived from the flesh of the snakehead fish (Channa striata). Before examining the ability of this non-serum albumin to promote capacitation in bovine sperm, the presence of albumin was detected using bromocresol purple (BCP) at the level of 25% from snakehead fish extract. Following the SDS-PAGE and densitometric analysis, two major bands at 40 kDa and 47 kDa consisting of 57% and 16% of total protein loaded were detected as the potential albumin-related bands. Significant differences were observed in all kinematic parameters upon incubation in the capacitation medium. Moreover, consistently higher values were shown for the kinematic parameters related to hyperactivation, such as amplitude lateral head (ALH), velocity curve linear (VCL), and linearity (LIN) when sperm were treated with 3 mg/mL of snakehead fish albumin among other treatments. Likewise, substantial differences of higher acrosome intact presented in sperm upon incubation with various concentrations of snakehead fish albumin for 90 minutes, indicating that this level of snakehead fish albumin can be used to replace the bovine serum albumin. However, further study is highly required to purify the albumin from snakehead fish extract for more reliable findings.

Keywords: capacitation promoter, snakehead fish, non-serum albumin, bovine sperm

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Authors: Sabyasachi Mukhopadhyay, Joseph Ogutu, Gundula Bartzke, Hans-Peter Piepho

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Rainfall is a critical component of climate governing vegetation growth and production, forage availability and quality for herbivores. However, reliable rainfall measurements are not always available, making it necessary to predict rainfall values for particular locations through time. Predicting rainfall in space and time can be a complex and challenging task, especially where the rain gauge network is sparse and measurements are not recorded consistently for all rain gauges, leading to many missing values. Here, we develop a flexible Bayesian model for predicting rainfall in space and time and apply it to Narok County, situated in southwestern Kenya, using data collected at 23 rain gauges from 1965 to 2015. Narok County encompasses the Maasai Mara ecosystem, the northern-most section of the Mara-Serengeti ecosystem, famous for its diverse and abundant large mammal populations and spectacular migration of enormous herds of wildebeest, zebra and Thomson's gazelle. The model incorporates geographical and meteorological predictor variables, including elevation, distance to Lake Victoria and minimum temperature. We assess the efficiency of the model by comparing it empirically with the established Gaussian process, Kriging, simple linear and Bayesian linear models. We use the model to predict total monthly rainfall and its standard error for all 5 * 5 km grid cells in Narok County. Using the Monte Carlo integration method, we estimate seasonal and annual rainfall and their standard errors for 29 sub-regions in Narok. Finally, we use the predicted rainfall to predict large herbivore biomass in the Maasai Mara ecosystem on a 5 * 5 km grid for both the wet and dry seasons. We show that herbivore biomass increases with rainfall in both seasons. The model can handle data from a sparse network of observations with many missing values and performs at least as well as or better than four established and widely used models, on the Narok data set. The model produces rainfall predictions consistent with expectation and in good agreement with the blended station and satellite rainfall values. The predictions are precise enough for most practical purposes. The model is very general and applicable to other variables besides rainfall.

Keywords: non-stationary covariance function, gaussian process, ungulate biomass, MCMC, maasai mara ecosystem

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Authors: Sudip Mondal, Biswanath Mondal, Sudit S. Mukhopadhyay, Apurba Dey

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Bioactive materials improve devices for a long lifespan but have mechanical limitations. Mechanical characterization is one of the very important characteristics to evaluate the life span and functionality of the scaffold material. After implantation of scaffold material the primary stage rejection of scaffold occurs due to non biocompatible effect of host body system. The second major problems occur due to the effect of mechanical failure. The mechanical and biocompatibility failure of the scaffold materials can be overcome by the prior evaluation of the scaffold materials. In this study chemically treated Labeo rohita scale is used for synthesizing hydroxyapatite (HAp) biomaterial. Thermo-gravimetric and differential thermal analysis (TG-DTA) is carried out to ensure thermal stability. The chemical composition and bond structures of wet ball-milled calcined HAp powder is characterized by Fourier Transform Infrared spectroscopy (FTIR), X-ray Diffraction (XRD), Field Emission Scanning Electron Microscopy (FE-SEM), Transmission Electron Microscopy (TEM), Energy Dispersive X-ray (EDX) analysis. Fish scale derived apatite materials consists of nano-sized particles with Ca/P ratio of 1.71. The biocompatibility through cytotoxicity evaluation and MTT assay are carried out in MG63 osteoblast cell lines. In the cell attachment study, the cells are tightly attached with HAp scaffolds developed in the laboratory. The result clearly suggests that HAp material synthesized in this study do not have any cytotoxic effect, as well as it has a natural binding affinity for mammalian cell lines. The synthesized HAp powder further successfully used to develop porous scaffold material with suitable mechanical property of ~0.8GPa compressive stress, ~1.10 GPa a hardness and ~ 30-35% porosity which is acceptable for implantation in trauma region for animal model. The histological analysis also supports the bio-affinity of processed HAp biomaterials in Wistar rat model for investigating the contact reaction and stability at the artificial or natural prosthesis interface for biomedical function. This study suggests the natural sourced fish scale-derived HAp material could be used as a suitable alternative biomaterial for tissue engineering application in near future.

Keywords: biomaterials, hydroxyapatite, scaffold, mechanical property, tissue engineering

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Authors: Avelyno D'Costa, S. K. Shyama, M. K. Praveen Kumar

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The coast of Goa, India receives constant anthropogenic stress through its major rivers which carry mining rejects of iron and manganese ores from upstream mining sites and petroleum hydrocarbons from shipping and harbor-related activities which put the aquatic fauna such as bivalves at risk. The present study reports the pollution status of the Goan coast by the above xenobiotics employing genotoxicity studies. This is further supplemented by the quantification of total petroleum hydrocarbons (TPHs) and various trace metals (iron, manganese, copper, cadmium, and lead) in gills of the estuarine clam, Meretrix ovum as well as from the surrounding water and sediment, over a two-year sampling period, from January 2013 to December 2014. Bivalves were collected from a probable unpolluted site at Palolem and a probable polluted site at Vasco, based upon the anthropogenic activities at these sites. Genotoxicity was assessed in the gill cells using the comet assay and micronucleus test. The quantity of TPHs and trace metals present in gill tissue, water and sediments were analyzed using spectrofluorometry and atomic absorption spectrophotometry (AAS), respectively. The statistical significance of data was analyzed employing Student’s t-test. The relationship between DNA damage and pollutant concentrations was evaluated using multiple regression analysis. Significant DNA damage was observed in the bivalves collected from Vasco which is a region of high industrial activity. Concentrations of TPHs and trace metals (iron, manganese, and cadmium) were also found to be significantly high in gills of the bivalves collected from Vasco compared to those collected from Palolem. Further, the concentrations of these pollutants were also found to be significantly high in the water and sediments at Vasco compared to that of Palolem. This may be due to the lack of industrial activity at Palolem. A high positive correlation was observed between the pollutant levels and DNA damage in the bivalves collected from Vasco suggesting the genotoxic nature of these pollutants. Further, M. ovum can be used as a bioindicator species for monitoring the level of pollution of the estuarine/coastal regions by TPHs and trace metals.

Keywords: comet assay, metals, micronucleus test, total petroleum Hydrocarbons

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Authors: Yunjia Yang, Peng Li, Gordon Xu, Timothy Mahony, Bing Zhang, Neena Mitter, Karishma Mody

Abstract:

Sheep flystrike is one of the most economically important diseases affecting the Australian sheep and wool industry (>356M/annually). Currently, control of Lucillia cuprina relies almost exclusively on chemicals controls and the parasite has developed resistance to nearly all control chemicals used in the past. It is therefore critical to develop an alternative solution for the sustainable control and management of flystrike. RNA interference (RNAi) technologies have been successfully explored in multiple animal industries for developing parasites controls. This research project aims to develop a RNAi based biological control for sheep blowfly. Double-stranded RNA (dsRNA) has already proven successful against viruses, fungi and insects. However, the environmental instability of dsRNA is a major bottleneck for successful RNAi. Bentonite polymer (BenPol) technology can overcome this problem, as it can be tuned for the controlled release of dsRNA in the gut challenging pH environment of the blowfly larvae, prolonging its exposure time to and uptake by target cells. To investigate the potential of BenPol technology for dsRNA delivery, four different BenPol carriers were tested for their dsRNA loading capabilities, and three of them were found to be capable of affording dsRNA stability under multiple temperatures (4°C, 22°C, 40°C, 55°C) in sheep serum. Based on stability results, dsRNA from potential targeted genes was loaded onto BenPol carriers and tested in larvae feeding assays, three genes resulting in knockdowns. Meanwhile, a primary blowfly embryo cell line (BFEC) derived from L. cuprina embryos was successfully established, aim for an effective insect cell model for testing RNAi efficacy for preliminary assessments and screening. The results of this study establish that the dsRNA is stable when loaded on BenPol particles, unlike naked dsRNA rapidly degraded in sheep serum. The stable nanoparticle delivery system offered by BenPol technology can protect and increase the inherent stability of dsRNA molecules at higher temperatures in a complex biological fluid like serum, providing promise for its future use in enhancing animal protection.

Keywords: flystrike, RNA interference, bentonite polymer technology, Lucillia cuprina

Procedia PDF Downloads 90

Authors: Vanessa Bennemann, Gabriela Laste, Márcia Inês Goettert

Abstract:

The gestational period is a phase in which the pregnant woman undergoes constant physiological and hormonal changes, which are part of the woman’s biological cycle, the development of the fetus, childbirth, and lactation. These are factors of response to the immunological adaptation of the human reproductive process that is directly related to the pregnancy’s well-being and development. Although most pregnancies occur without complications, about 15% of pregnant women will develop potentially fatal complications, implying maternal and fetal risk. Therefore, requiring specialized care for high-risk pregnant women (HRPW) with obstetric interventions for the survival of the mother and/or fetus. Among the risk factors that characterize HRPW are the women's age, gestational diabetes mellitus (GDM), autoimmune diseases, infectious diseases such as syphilis and HIV, hypertension (SAH), preeclampsia, eclampsia, HELLP syndrome, uterine contraction abnormalities, and premature placental detachment (PPD), thyroid disorders, among others. Thus, pregnancy has an impact on the thyroid gland causing changes in the functioning of the mother's thyroid gland, altering the thyroid hormone (TH) profiles and production as pregnancy progresses. Considering, throughout the gestational period, the interpretation of the results of the tests to evaluate the thyroid functioning depends on the stage in which the pregnancy is. Thyroid disorders are directly related to adverse obstetric outcomes and in child development. Therefore, the adequate release of TH is important for a pregnancy without complications and optimal fetal growth and development. Objective: Investigate the physiological effects caused by thyroid disorders in the gestational period. Methods: A search for articles indexed in PubMed, Scielo, and MDPI databases, was performed using the term “AND”, with the descriptors: Pregnancy, Thyroid. With several combinations that included: Melatonin, Thyroidopathy, Inflammatory processes, Cytokines, Anti-inflammatory, Antioxidant, High-risk pregnancy. Subsequently, the screening was performed through the analysis of titles and/or abstracts. The criteria were: including clinical studies in general, randomized or not, in the period of 10 years prior to the research, in the English literature; excluded: experimental studies, case reports, research in the development phase. Results: In the preliminary results, a total of studies (n=183) were found, (n=57) excluded, such as studies of cancer, diabetes, obesity, and skin diseases. Conclusion: To date, it has been identified that thyroid diseases can impair the fetus’s brain development. Further research is suggested on this matter to identify new substances that may have a potential therapeutic effect to aid the gestational period with thyroid diseases.

Keywords: pregnancy, thyroid, melatonin, high-risk pregnancy

Procedia PDF Downloads 141