Search results for: Anatolian wild goat
201 Chemical Analysis and Cytotoxic Evaluation of Asphodelus Aestivus Brot. Flowers
Authors: Mai M. Farid, Mona El-Shabrawy, Sameh R. Hussein, Ahmed Elkhateeb, El-Said S. Abdel-Hameed, Mona M. Marzouk
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Asphodelus aestivus Brot. Is a wild plant distributed in Egypt and is considered one of the five Asphodelus spp. from the family Asphodelaceae; it grows in dry grasslands and on rocky or sandy soil. The chemical components of A. aestivus flowers extract were analyzed using different chromatographic and spectral techniques and led to the isolation of two anthraquinones identified as emodin and emodin-O-glucoside. In addition to, five flavonoid compounds;kaempferol,Kaempferol-3-O-glucoside,Apigenin-6-C-glucoside-7-O-glucoside (Saponarine), luteolin 7-O-β-glucopyranoside, Isoorientin-O-malic acid which is a new compound in nature. The LC-ESI-MS/MS analysis of the flower extract of A. aestivus led to the identification of twenty- two compounds characterized by the presence of flavones, flavonols, and flavone C-glycosides. While GC/MS analysis led to the identification of 24 compounds comprising 98.32% of the oil, the major components of the oil were 9, 12, 15-Octadecatrieoic acid methyl ester 28.72%, and 9, 12-Octadecadieroic acid (Z, Z)-methyl ester 19.96%. In vitro cytotoxic activity of the aqueous methanol extract of A. aestivus flowers against HEPG2, HCT-116, MCF-7, and A549 culture was examined and showed moderate inhibition (62.3±1.1)% on HEPG2 cell line followed by (36.8±0.2)% inhibition on HCT-116 and a weak inhibition (5.7± 0.0.2) on MCF-7 cell line followed by (4.5± 0.4) % inhibition on A549 cell line and this is considered the first cytotoxic report of A. aestivus flowers.Keywords: Anthraquinones, Asphodelus aestivus, Cytotoxic activity, Flavonoids, LC-ESI-MS/MS
Procedia PDF Downloads 223200 Impact of El-Matrouha Landfill on Oued El-Kebir (North East of Algeria)
Authors: Mohamed Djalil Zaafour, Samir Chekchaki, Mohamed Benslama
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The Landfill of El Matrouha is located in El-Tarf town (extreme north east of Algeria), the Landfill is present as a gigantic wild dump. This waste dump occupies an area of over four hectares, tons of rubbish that is sent daily are scattered over kilometers, reaching farmland located west of the town, the landfill is close to a temporary Oued, which supply Oued Guergour the last tributary Oued El Kebir. The landfills are causing serious environmental damage, following the infiltration of leachates, which contribute to the degradation of water quality, in the context of this problem, the purpose of the work is focused on assessing the impact of this landfill on Oued El-Kebir, for this a series of sampling and analysis of the soil and water of this Oued was performed; The results show that the soil collected reveal the sandy texture facilitating infiltration and percolation of leachate from the landfill; the physicochemical analysis of the quality of the river water reveals high levels of sulfates in fact this element is one of the essential constituents of the mineral fraction of the waste presenting a risk of pollution by this element, The recorded values for nutrients are sub-standard, for trace elements analysis shows very low metal load on the river except for lead, which is present at high concentrations exceeding all standard.Keywords: Algeria, landfill, leachates, Oued El-kebir
Procedia PDF Downloads 372199 The Role of Inflammasomes for aβ Microglia Phagocytosis in Alzheimer Disease
Authors: Francesca La Rosa , Marina Saresella, Mario Clerici, Michael Heneka
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Neuroinflammation plays a key role in the modulation of the pathogenesis of neurodegenerative disorder such as Alzheimer's Disease (AD). Microglia, the main immune effector of the brain, are able to migrate to sites of Amyloid-beta (Aβ) deposition to eliminate Aβ phagocytosis upon activation by multiple receptors: Toll like receptors and scavenger receptors. The issue of whether microglia are able to eliminate pathological lesions such as neurofibrillary tangles or senile plaques from AD brain still remains the matter of controversy. Recent data suggest that the Nod Like Receptor 3 (NLRP3), multiprotein inflammasome complexes, plays a role in AD, as its activation in the microglia by Aβ triggers. IL-1β is produced as a biologically inactive pro-form and requires caspase-1 for activation and secretion. Caspase-1 activity is controlled by inflammasomes. We investigate about the importance of inflammasomes complex in the Aβ phagocytosis and its degradation. The preliminary results of phagocytosis assay and immunofluorescent experiment on primary Microglia cells to lipopolysaccharide (LPS) an Aβ exposure show that a previous treatment with LPS reduce Aβ phagocytosis. Different results were obtained in Primary Microglia wild type, NLRP3 and ASC Knockout suggesting a real inflammasomes involvement in Alzheimer's pathology. Inflammasomes inactivation reduces the production of inflammatory cytokines prolonging the protective activity of microglia and Aβ clearance, featuring a typical microglia phenotype of the early stage of AD disease.Keywords: Alzheimer disease, innate immunity, neuroinflammation, NLRP3
Procedia PDF Downloads 457198 Protective Efficacy of Curcuma Aromatica Leaf Extract on Liver of Arsenic Intoxicated Albino Rats
Authors: Priya Bajaj, Baby Tabassum
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Arsenic is a poisonous metalloid, naturally occurring in soil, air, rocks and ground water. This dreadful metalloid commonly exists as inorganic compound, arsenic trioxide. WHO permitted maximum limit for arsenic in water is 0.01 mg/L, but some affected areas show ground water level of arsenic up to 3 mg/L even. Ground water arsenic pollution has created a number of health problems, viz. keratosis, melanosis, lesions and even skin cancers. The key objective of our nested study was to characterize arsenic induced hepatotoxicity and to find out some herbal protection against it. For the purpose, we selected albino rat (Rattus norvegicus) as model for arsenic induced liver injury and wild turmeric (Curcuma aromatica) leaf extract as remedy for it. The study was performed at acute (1 day) and subacute (7, 14 & 21 days) levels. The LD50 estimated for arsenic trioxide was 14.98 mg/kg body weight. In our investigation, we observed a significant restoration of altered hepatic lipid, cholesterol, protein and glycogen contents as well as liver weight, body-weight and hepato-somatic index by Curcuma aromatica leaf extract before arsenic intoxication. The results reveal excellent protective efficacy of Curcuma aromatica leaf extract that further can be exploited in remediation programme in heavy metal affected areas.Keywords: arsenic, Curcuma aromatica, glycogen, lipids
Procedia PDF Downloads 255197 ParkedGuard: An Efficient and Accurate Parked Domain Detection System Using Graphical Locality Analysis and Coarse-To-Fine Strategy
Authors: Chia-Min Lai, Wan-Ching Lin, Hahn-Ming Lee, Ching-Hao Mao
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As world wild internet has non-stop developments, making profit by lending registered domain names emerges as a new business in recent years. Unfortunately, the larger the market scale of domain lending service becomes, the riskier that there exist malicious behaviors or malwares hiding behind parked domains will be. Also, previous work for differentiating parked domain suffers two main defects: 1) too much data-collecting effort and CPU latency needed for features engineering and 2) ineffectiveness when detecting parked domains containing external links that are usually abused by hackers, e.g., drive-by download attack. Aiming for alleviating above defects without sacrificing practical usability, this paper proposes ParkedGuard as an efficient and accurate parked domain detector. Several scripting behavioral features were analyzed, while those with special statistical significance are adopted in ParkedGuard to make feature engineering much more cost-efficient. On the other hand, finding memberships between external links and parked domains was modeled as a graph mining problem, and a coarse-to-fine strategy was elaborately designed by leverage the graphical locality such that ParkedGuard outperforms the state-of-the-art in terms of both recall and precision rates.Keywords: coarse-to-fine strategy, domain parking service, graphical locality analysis, parked domain
Procedia PDF Downloads 409196 Visualizing Matrix Metalloproteinase-2 Activity Using Extracellular Matrix-Immobilized Fluorescence Resonance Energy Transfer Bioprobe in Cancer Cells
Authors: Hawon Lee, Young-Pil Kim
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Visualizing matrix metalloproteinases (MMPs) activity is necessary for understanding cancer metastasis because they are implicated in cell migration and invasion by degrading the extracellular matrix (ECM). While much effort has been made to sense the MMP activity, but extracellularly long-term monitoring of MMP activity still remains challenging. Here, we report a collagen-bound fluorescent bioprobe for the detection of MMP-2 activity in the extracellular environment. This bioprobe consists of ECM-immobilized part (including collagen-bound protein) and MMP-sensing part (including peptide substrate linked with fluorescence resonance energy transfer (FRET) coupler between donor green fluorescent protein (GFP) and acceptor TAMRA dye), which was constructed through intein-mediated self-splicing conjugation. Upon being immobilized on the collagen-coated surface, this bioprobe enabled efficient long-lasting observation of MMP-2 activity in the cultured cells without affecting cell growth and viability. As a result, the FRET ratio (acceptor/donor) decreased as the MMP2 activity increased in cultured cancer cells. Furthermore, unlike wild-type MMP-2, mutated MMP-2 expression (Y580A in the hemopexin region) gave rise to lowering the secretion of MMP-2 in HeLa. Conclusively, our method is anticipated to find applications for tracing and visualizing enzyme activity.Keywords: collagen, ECM, FRET, MMP
Procedia PDF Downloads 203195 Characterization of Fatty Acid Glucose Esters as Os9BGlu31 Transglucosidase Substrates in Rice
Authors: Juthamath Komvongsa, Bancha Mahong, Kannika Phasai, Sukanya Luang, Jong-Seong Jeon, James Ketudat-Cairns
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Os9BGlu31 is a rice transglucosidase that transfers glucosyl moieties to various acceptors such as carboxylic acids and alcohols, including phenolic acids and flavonoids, in vitro. The role of Os9BGlu31 transglucosidase in rice plant metabolism has not been reported to date. Methanolic extracts of rice bran and flag leaves were found to contain substrates to which Os9BGlu31 could transfer glucose from 4-nitrophenyl β -D-glucopyranoside donor. The semi-purified substrate from rice bran was found to contain oleic acid and linoleic acid and the pure fatty acids were found to act as acceptor substrates for Os9BGlu31 transglucosidase to form 1-O-acyl glucose esters. Os9BGlu31 showed higher activity with oleic acid (18:1) and linoleic acid (18:2) than stearic acid (18:0), and had both higher kcat and higher Km for linoleic than oleic acid in the presence of 8 mM 4NPGlc donor. This transglucosidase reaction is reversible, Os9bglu31 knockout rice lines of flag leaves were found to have higher amounts of fatty acid glucose esters than wild type control lines, these data conclude that fatty acid glucose esters act as glucosyl donor substrates for Os9BGlu31 transglucosidase in rice.Keywords: fatty acid, fatty acid glucose ester, transglucosidase, rice flag leaf, homologous knockout lines, tandam mass spectrometry
Procedia PDF Downloads 368194 Seasonal Variation of the Essential Oils of Foeniculum vulgare Miller and Carum carvi L. Cultivated in Algerian Sahara
Authors: K. Fyad, A. Cheriti, Y. Bourmita, N. Belboukhari
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Many industries are involved by using essential oils such as food, flavour, and beverage, pharmaceutical, cosmetic and fragrance. Apiaceae species are usually herbs, rarely schrubs characterized particularly by its inflorescence typical umbel. Many species of this family have been widely used in folk medicine throughout the world. The most characteristic natural compounds in this family are the essential oils secreted in schizogenous canals in all organs with remarkable variability chemical composition. As a part of our investigation into medicinal plants growing in Algerian Sahara. In this study, we investigate the chemical composition of the essential oils extracted from two Apiaceae species: Foeniculum vulgare Miller and Carum carvi L cultivated in the Sahara. The plants were selected on the basis of their use by local people to treat infectious diseases as determined in our previous ethnopharmacological study. Wild samples of Foeniculum vulgare Miller and Carum carvi L cultivated in an experimental field at the university. The harvest was made during the year 2011 according to the growth cycle stage of the plants. The essential oils of different fresh aerial parts, obtained by hydrodistillation were analysed by GC. The results showed that the essential oils yields are not uniform among the different cycle stage. The percentage of components is significantly affected by the harvesting period of the plant material.Keywords: essential oils, Apiaceae, growth cycle, Sahara, GC
Procedia PDF Downloads 432193 Experimental Investigation of Cutting Forces and Temperature in Bone Drilling
Authors: Vishwanath Mali, Hemant Warhatkar, Raju Pawade
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Drilling of bone has been always challenging for surgeons due to the adverse effect it may impart to bone tissues. Force has to be applied manually by the surgeon while performing conventional bone drilling which may lead to permanent death of bone tissues and nerves. During bone drilling the temperature of the bone tissues increases to higher values above 47 ⁰C that causes thermal osteonecrosis resulting into screw loosening and subsequent implant failures. An attempt has been made here to study the input drilling parameters and surgical drill bit geometry affecting bone health during bone drilling. A One Factor At a Time (OFAT) method is used to plan the experiments. Input drilling parameters studied include spindle speed and feed rate. The drill bit geometry parameter studied include point angle and helix angle. The output variables are drilling thrust force and bone temperature. The experiments were conducted on goat femur bone at room temperature 30 ⁰C. For measurement of thrust forces KISTLER cutting force dynamometer Type 9257BA was used. For continuous data acquisition of temperature NI LabVIEW software was used. Fixture was made on RPT machine for holding the bone specimen while performing drilling operation. Bone specimen were preserved in deep freezer (LABTOP make) under -40 ⁰C. In case of drilling parameters, it is observed that at constant feed rate when spindle speed increases, thrust force as well as temperature decreases and at constant spindle speed when feed rate increases thrust force as well as temperature increases. The effect of drill bit geometry shows that at constant helix angle when point angle increases thrust force as well as temperature increases and at constant point angle when helix angle increase thrust force as well as temperature decreases. Hence it is concluded that as the thrust force increases temperature increases. In case of drilling parameter, the lowest thrust force and temperature i.e. 35.55 N and 36.04 ⁰C respectively were recorded at spindle speed 2000 rpm and feed rate 0.04 mm/rev. In case of drill bit geometry parameter, the lowest thrust force and temperature i.e. 40.81 N and 34 ⁰C respectively were recorded at point angle 70⁰ and helix angle 25⁰ Hence to avoid thermal necrosis of bone it is recommended to use higher spindle speed, lower feed rate, low point angle and high helix angle. The hard nature of cortical bone contributes to a greater rise in temperature whereas a considerable drop in temperature is observed during cancellous bone drilling.Keywords: bone drilling, helix angle, point angle, thrust force, temperature, thermal necrosis
Procedia PDF Downloads 310192 Molecular Detection of Viruses Causing Hemorrhagic Fevers in Rodents in the South-West of Korea
Authors: Sehrish Jalal, Choon-Mee Kim, Dong-Min Kim
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Background: Many pathogens causing hemorrhagic fevers of medical and veterinary importance have been identified and isolated from rodents in the Republic of Korea (ROK). Objective: We investigated the prevalence of emerging viruses causing hemorrhagic fevers, such as hemorrhagic fever with renal syndrome (HFRS), severe fever with thrombocytopenia syndrome (SFTS) and flaviviruses, from wild rodents. Methods: Striped field mice, Apodemus agrarius, (n=39) were captured during 2014-2015 in the south-west of ROK. Using molecular methods, lung samples were evaluated for SFTS virus, HFRS virus and flavivirus, and seropositivity was evaluated in the blood. Results: A high positive rate of Hantavirus (46.2%) was detected in A.agrarius lungs by reverse transcription-nested polymerase chain reaction (RT-N-PCR). The monthly prevalence of HFRS virus was 16.7% in October, 86.7% in November and 25% in August of the following year (p < 0.001). Moreover, 17.9% of blood samples were serologically positive for Hantavirus antibodies. The most prevalent strain in A. agrarius was Hantaan virus. All samples were positive for neither SFTS nor flavivirus. Conclusion: Hantan virus was detected in 86.7% of A. agrarius in November (autumn), and thus, virus shedding from A. agrarius can increase the risk of humans contracting HFRS. These findings may help to predict and prevent disease outbreaks in ROK.Keywords: hemorrhagic fever virus, molecular diagnostic technique, rodents, Korea
Procedia PDF Downloads 161191 Effects of Fire on Vegetation of the Prairies and Black Oak Sand Savannas of Kankakee, Illinois
Authors: Megan Alkazoff, Charles Ruffner
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Tallgrass prairies and sand savannas, once covering northern to central Illinois, are ecosystems in need of restoration and conservation in the Midwestern United States. The Nature Conservancy manages five sites containing fragments of remaining tallgrass prairies and sand savannas within the Kankakee Sands using techniques such as prescribed burning and invasive species removal. The objective of this study was to conduct a ten-year resampling of transects established on these five sites during previous studies to assess whether the management tools applied there are helping maintain the tallgrass prairie and sand savannas. During the summer of 2020, permanent transect lines were sampled using a quadrat to determine the % Cover Class of each species rooted in the quadrat. Data gathered was analyzed using linear regression to illustrate the relationship between fire occurrence and species composition on the landscape. The fire frequency had a highly significant effect (P= 0.0025) on the species richness of all sites. The frequency of fire had a non-significant effect (P>0.05) on the Floristic Quality Index, percent C value 4-10, and bare-ground percentage of a site. These results suggest that fire on the landscape, both wild and prescribed, have increased biodiversity on all five sites but has not affected the Floristic Quality Index, percent C value 4-10, and the percentage of bare-ground on the sites.Keywords: fire, floristic quality assessment, sand savanna, species richness, tallgrass prairie
Procedia PDF Downloads 179190 The Application of Animal Welfare Certification System for Farm Animal in South Korea
Authors: Ahlyum Mun, Ji-Young Moon, Moon-Seok Yoon, Dong-Jin Baek, Doo-Seok Seo, Oun-Kyong Moon
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There is a growing public concern over the standards of farm animal welfare, with higher standards of food safety. In addition, the recent low incidence of Avian Influenza in laying hens among certificated farms is receiving attention. In this study, we introduce animal welfare systems covering the rearing, transport and slaughter of farm animals in South Korea. The concepts of animal welfare farm certification are based on ensuring the five freedoms of animal. The animal welfare is also achieved by observing the condition of environment including shelter and resting area, feeding and water and the care for the animal health. The certification of farm animal welfare is handled by the Animal Protection & Welfare Division of Animal and Plant Quarantine Agency (APQA). Following the full amendment of Animal Protection Law in 2011, animal welfare farm certification program has been implemented since 2012. The certification system has expanded to cover laying hen, swine, broiler, beef cattle and dairy cow, goat and duck farms. Livestock farmers who want to be certified must apply for certification at the APQA. Upon receipt of the application, the APQA notifies the applicant of the detailed schedule of the on-site examination after reviewing the document and conducts the on-site inspection according to the evaluation criteria of the welfare standard. If the on-site audit results meet the certification criteria, APQA issues a certificate. The production process of certified farms is inspected at least once a year for follow-up management. As of 2017, a total of 145 farms have been certified (95 laying hen farms, 12 swine farms, 30 broiler farms and 8 dairy cow farms). In addition, animal welfare transportation vehicles and slaughterhouses have been designated since 2013 and currently 6 slaughterhouses have been certified. Animal Protection Law has been amended so that animal welfare certification marks can be affixed only to livestock products produced by animal welfare farms, transported through animal welfare vehicles and slaughtered at animal welfare slaughterhouses. The whole process including rearing–transportation- slaughtering completes the farm animal welfare system. APQA established its second 5-year animal welfare plan (2014-2019) that includes setting a minimum standard of animal welfare applicable to all livestock farms, transportation vehicles and slaughterhouses. In accordance with this plan, we will promote the farm animal welfare policy in order to truly advance the Korean livestock industry.Keywords: animal welfare, farm animal, certification system, South Korea
Procedia PDF Downloads 401189 Effect of Manure Treatment on Furrow Erosion: A Case Study of Sagawika Irrigation Scheme in Kasungu, Malawi
Authors: Abel Mahowe
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Furrow erosion is the major problem menacing sustainability of irrigation in Malawi and polluting water bodies resulting in death of many aquatic animals. Many rivers in Malawi are drying due to some poor practices that are being practiced around these water bodies, furrow erosion is one of the cause of sedimentation in these rivers although it has gradual effect on deteriorating of these rivers hence neglected, but has got long term disastrous effect on water bodies. Many aquatic animals also suffer when these sediments are taken into these water bodies. An assessment of effect of manure treatment on furrow erosion was carried out in Sagawika irrigation scheme located in Kasungu District north part of Malawi. The soil on the field was clay loam and had just been tilled. The average furrow slope of 0.2% and was divided into two blocks, A and B. Each block had 20V-shaped furrow having a length of 10 m. Three different manure were used to construct these furrows by mixing it with soil which was moderately moist and 5 furrows from each block were constructed without manure. In each block 5furrow were made using a specific type of manure, and one set of five furrows in each block was made without manure treatment. The types of manure that were used were goat manure, pig manure, and manure from crop residuals. The manure application late was 5 kg/m. The furrow was constructed at a spacing of 0.6 m. Tomato was planted in the two blocks at spacing of 0.15 m between rows and 0.15 m between planting stations. Irrigation water was led from feeder canal into the irrigation furrows using siphons. The siphons discharge into each furrow was set at 1.86 L/S. The ¾ rule was used to determine the cut-off time for the irrigation cycles in order to reduce the run-off at the tail end. During each irrigation cycle, samples of the runoff water were collected at one-minute intervals and analyzed for total sediment concentration for use in estimating the total soil sediment loss. The results of the study have shown that a significant amount of soil is lost in soils without many organic matters, there was a low level of erosion in furrows that were constructed using manure treatment within the blocks. In addition, the results have shown that manure also differs in their ability to control erosion since pig manure proved to have greater abilities in binding the soil together than other manure since they were reduction in the amount of sediments at the tail end of furrows constructed by this type of manure. The results prove that manure contains organic matters which helps soil particles to bind together hence resisting the erosive force of water. The use of manure when constructing furrows in soil with less organic matter can highly reduce erosion hence reducing also pollution of water bodies and improve the conditions of aquatic animals.Keywords: aquatic, erosion, furrow, soil
Procedia PDF Downloads 287188 The Role Collagen VI Plays in Heart Failure: A Tale Untold
Authors: Summer Hassan, David Crossman
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Myocardial fibrosis (MF) has been loosely defined as the process occurring in the pathological remodeling of the myocardium due to excessive production and deposition of extracellular matrix (ECM) proteins, including collagen. This reduces tissue compliance and accelerates progression to heart failure, as well as affecting the electrical properties of the myocytes resulting in arrhythmias. Microscopic interrogation of MF is key to understanding the molecular orchestrators of disease. It is well-established that recruitment and stimulation of myofibroblasts result in Collagen deposition and the resulting expansion in the ECM. Many types of Collagens have been identified and implicated in scarring of tissue. In a series of experiments conducted at our lab, we aim to elucidate the role collagen VI plays in the development of myocardial fibrosis and its direct impact on myocardial function. This was investigated through an animal experiment in Rats with Collagen VI knockout diseased and healthy animals as well as Collagen VI wild diseased and healthy rats. Echocardiogram assessments of these rats ensued at four-time points, followed by microscopic interrogation of the myocardium aiming to correlate the role collagen VI plays in myocardial function. Our results demonstrate a deterioration in cardiac function as represented by the ejection fraction in the knockout healthy and diseased rats. This elucidates a potential protective role that collagen-VI plays following a myocardial insult. Current work is dedicated to the microscopic characterisation of the fibrotic process in all rat groups, with the results to follow.Keywords: heart failure, myocardial fibrosis, collagen, echocardiogram, confocal microscopy
Procedia PDF Downloads 83187 Microglia Activation in Animal Model of Schizophrenia
Authors: Esshili Awatef, Manitz Marie-Pierre, Eßlinger Manuela, Gerhardt Alexandra, Plümper Jennifer, Wachholz Simone, Friebe Astrid, Juckel Georg
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Maternal immune activation (MIA) resulting from maternal viral infection during pregnancy is a known risk factor for schizophrenia. The neural mechanisms by which maternal infections increase the risk for schizophrenia remain unknown, although the prevailing hypothesis argues that an activation of the maternal immune system induces changes in the maternal-fetal environment that might interact with fetal brain development. It may lead to an activation of fetal microglia inducing long-lasting functional changes of these cells. Based on post-mortem analysis showing an increased number of activated microglial cells in patients with schizophrenia, it can be hypothesized that these cells contribute to disease pathogenesis and may actively be involved in gray matter loss observed in such patients. In the present study, we hypothesize that prenatal treatment with the inflammatory agent Poly(I:C) during embryogenesis at contributes to microglial activation in the offspring, which may, therefore, represent a contributing factor to the pathogenesis of schizophrenia and underlines the need for new pharmacological treatment options. Pregnant rats were treated with intraperitoneal injections a single dose of Poly(I:C) or saline on gestation day 17. Brains of control and Poly(I:C) offspring, were removed and into 20-μm-thick coronal sections were cut by using a Cryostat. Brain slices were fixed and immunostained with ba1 antibody. Subsequently, Iba1-immunoreactivity was detected using a secondary antibody, goat anti-rabbit. The sections were viewed and photographed under microscope. The immunohistochemical analysis revealed increases in microglia cell number in the prefrontal cortex, in offspring of poly(I:C) treated-rats as compared to the controls injected with NaCl. However, no significant differences were observed in microglia activation in the cerebellum among the groups. Prenatal immune challenge with Poly(I:C) was able to induce long-lasting changes in the offspring brains. This lead to a higher activation of microglia cells in the prefrontal cortex, a brain region critical for many higher brain functions, including working memory and cognitive flexibility. which might be implicated in possible changes in cortical neuropil architecture in schizophrenia. Further studies will be needed to clarify the association between microglial cells activation and schizophrenia-related behavioral alterations.Keywords: Microglia, neuroinflammation, PolyI:C, schizophrenia
Procedia PDF Downloads 417186 Sterols Regulate the Activity of Phospholipid Scramblase by Interacting through Putative Cholesterol Binding Motif
Authors: Muhasin Koyiloth, Sathyanarayana N. Gummadi
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Biological membranes are ordered association of lipids, proteins, and carbohydrates. Lipids except sterols possess asymmetric distribution across the bilayer. Eukaryotic membranes possess a group of lipid translocators called scramblases that disrupt phospholipid asymmetry. Their action is implicated in cell activation during wound healing and phagocytic clearance of apoptotic cells. Cholesterol is one of the major membrane lipids distributed evenly on both the leaflet and can directly influence the membrane fluidity through the ordering effect. The fluidity has an impact on the activity of several membrane proteins. The palmitoylated phospholipid scramblases localized to the lipid raft which is characterized by a higher number of sterols. Here we propose that cholesterol can interact with scramblases through putative CRAC motif and can modulate their activity. To prove this, we reconstituted phospholipid scramblase 1 of C. elegans (SCRM-1) in proteoliposomes containing different amounts of cholesterol (Liquid ordered/Lo). We noted that the presence of cholesterol reduced the scramblase activity of wild-type SCRM-1. The interaction between SCRM-1 and cholesterol was confirmed by fluorescence spectroscopy using NBD-Chol. Also, we observed loss of such interaction when one of I273 in the CRAC motif mutated to Asp. Interestingly, the point mutant has partially retained scramblase activity in Lo vesicles. The current study elucidated the important interaction between cholesterol and SCRM-1 to fine-tune its activity in artificial membranes.Keywords: artificial membranes, CRAC motif, plasma membrane, PL scramblase
Procedia PDF Downloads 175185 Growing Evaluation Process in Chamaedorea Linearis with Humus from Biosolids of the Wastewater Treatment Plant, Nueva Granada Military University Cajica
Authors: J. Gonzalez, P. Jimenez, C. Isaza
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Palms have different characteristics that make them vulnerable; that is the case of the Chamaedorea linearis, with the presence of solitary stems of small diameter and medium leaves, culturally harvested, and in religious festivities used. Additionally, they present a weak apical meristem as the only emergency point, slow development and growth, and an affectation due to the high rate of deforestation in Colombia. Propagation of this species can improve the pressure on wild populations and help their survival in the environment. In this study was used in 177 plants biosolids humus from the Wastewater Treatment Plant (WWTP), located at the UMNG Campus Cajica (Cundinamarca, Colombia). The experiment used a control and two treatments with 10% and 20% of humus. During the process, the variables evaluated were number of leaves, percentage of chlorophyll, stem length, and estimated leaf area. The data set were taking during 14 weeks before the reproductive maturity, evidencing that the most representative development of the palms was in the treatment of 20%, plants in this treatment presented major number of leaves, larger stems, a high quantity of chlorophyll, and was a first treatment that present pinnate leaves them represent an important point in maturity process. The research gives an opportunity to improve times of growth in another species of palms and plants (Product result from INV ING 2986 UMNG).Keywords: biosolids, humus, growth, palms, wastewater treatment plant, WWTP
Procedia PDF Downloads 126184 Atomic Town: History and Vernacular Heritage at the Mary Kathleen Uranium Mine in Australia
Authors: Erik Eklund
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Mary Kathleen was a purpose-built company town located in northwest Queensland in Australia. It was created to work on a rich uranium deposit discovered in the area in July 1954. The town was complete by 1958, possessing curved streets, modern materials, and a progressive urban planning scheme. Formed in the minds of corporate executives and architects and made manifest in arid zone country between Cloncurry and Mount Isa, Mary Kathleen was a modern marvel in the outback, a town that tamed the wild country of northwest Queensland, or so it seemed. The town was also a product of the Cold War. In the context of a nuclear arms race between the Soviet Union and her allies, and the United States of America (USA) and her Allies, a rapid rush to locate, mine, and process uranium after 1944 led to the creation of uranium towns in Czechoslovakia, Canada, the Soviet Union, USA and Australia of which Mary Kathleen was one such example. Mary Kathleen closed in 1981, and most of the town’s infrastructure was removed. Since then, the town’s ghostly remains have attracted travellers and tourists. Never an officially-sanctioned tourist site, the area has nevertheless become a regular stop for campers and day trippers who have engaged with the site often without formal interpretation. This paper explores the status of this vernacular heritage and asks why it has not gained any official status and what visitors might see in the place despite its uncertain status.Keywords: uranium mining, planned communities, official heritage, vernacular heritage, Australian history
Procedia PDF Downloads 91183 Characterization of Some Bread Wheat Genotypes for Drought Tolerance Using Molecular Markers
Authors: Begüm Terzi, Özlem Ateş Sönmezoğlu, Ahmet Yildirim
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Drought is the most important factor that limiting the production and productivity of wheat in the world. The yield of wheat, which is one of the most important crop in the world, reduced depend on drought. Researches to minimize effects of drought are one of the most important about breeding of drought resistant varieties. In recent years, benefiting from the drought resistance wild species and rapid advances in molecular biology studies, researches about drought have been accelerated and number of studies were made on molecular plant breeding which included the molecular mechanisms related to drought resistance. The aim of the present study was characterization of some bread wheat lines for drought tolerance which commonly cultivated in different location of Turkey. In this study, registered 9 bread wheat varieties which on the physiological tests about drought tolerance and 10 bread wheat line has been developed by Transitional Zone Agricultural Research Institute were used. SSR, STS, RAPD and SNP markers that associated with drought tolerance were used. The polymorphisms of the markers were determined by screening of two control varieties. For these purpose 40 molecular markers were used and 12 markers of them were polymorphic among the drought tolerance and the drought sensitive varieties. Control varieties were screened using polymorphic markers. All the DNAs on the genotypes will be searched for the presence of QTLs mapped to different chromosomes. Result of the research, the studied genotypes will be grouped according to drought tolerance and will be detected drought tolerance varieties by molecular markers. In addition, the results will be compared also with physiological tests. The drought tolerant wheat genotypes may be used in breeding studies related to drought stress.Keywords: bread wheat, drought, molecular marker, Triticum aestivum
Procedia PDF Downloads 386182 Genetic Diversity and Variation of Nigerian Pigeon (Columba livia domestica) Populations Based on the Mitochondrial Coi Gene
Authors: Foluke E. Sola-Ojo, Ibraheem A. Abubakar, Semiu F. Bello, Isiaka H. Fatima, Sule Bisola, Adesina M. Olusegun, Adeniyi C. Adeola
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The domesticated pigeon, Columba livia domestica, has many valuable characteristics, including high nutritional value and fast growth rate. There is a lack of information on its genetic diversity in Nigeria; thus, the genetic variability in mitochondrial cytochrome oxidase subunit I (COI) sequences of 150 domestic pigeons from four different locations was examined. Three haplotypes (HT) were identified in Nigerian populations; the most common haplotype, HT1, was shared with wild and domestic pigeons from Europe, America, and Asia, while HT2 and HT3 were unique to Nigeria. The overall haplotype diversity was 0.052± 0.025, and nucleotide diversity was 0.026± 0.068 across the four investigated populations. The phylogenetic tree showed significant clustering and genetic relationship of Nigerian domestic pigeons with other global pigeons. The median-joining network showed a star-like pattern suggesting population expansion. AMOVA results indicated that genetic variations in Nigerian pigeons mainly occurred within populations (99.93%), while the Neutrality tests results suggested that the Nigerian domestic pigeons’ population experienced recent expansion. This study showed a low genetic diversity and population differentiation among Nigerian domestic pigeons consistent with a relatively conservative COI sequence with few polymorphic sites. Furthermore, the COI gene could serve as a candidate molecular marker to investigate the genetic diversity and origin of pigeon species. The current data is insufficient for further conclusions; therefore, more research evidence from multiple molecular markers is required.Keywords: Nigeria pigeon, COI, genetic diversity, genetic variation, conservation
Procedia PDF Downloads 201181 Phytochemistry and Biological Activity of Extracts of the Red Raspberry Rubus rosifolius
Authors: Theresa Campbell, Camille Bowen-Forbes, William Aalbersberg
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Differences in the sensory properties of two subtly distinct varieties of Rubus rosifolius lead to the examination of their anthocyanin, essential oil and polyphenol profiles. In both cases, notable differences were identified. Pelargonidin-3-rhutinoside (17.2 mg/100 g FW) and Cyanidin-3-glucoside (66.2 mg/100g FW) proved to be the dominant anthocyanins in the red and wine red varieties respectively. Linalool and terpineol were the major constituents of the essential oil from the red variety; however, those of the wine red variety are unidentified. In regard to phenolic compounds, caffeic acid and quercetin were in a higher concentration in the red variety (1.85 and 0.73 mg/100g FW respectively, compared to 1.22 and 0.34 mg/100g FW respectively in the wine red fruits); while ellagic acid and ferulic acid were of a higher concentration in the wine red variety (0.92 and 0.84mg/100g FW respectively, compared to 0.15 and 0.48 mg/100g FW respectively in the red variety). The methanol extract of both fruit varieties showed great antioxidant activity. Analysis of the antimicrobial activity of the fruit extracts against the growth of drug resistant pathogens revealed that they are active against methicillin resistant S. aureus (MRSA), rifampicin resistant S. aureus (RRSA), wild-type S. aureus (WTSA) and vancomycin-resistant Enterococcus faecium (VREF). Activity was also reported against several food-borne pathogens including two strains of E. coli, L. monocytogenes and Enterobacter aerogenes. The cytotoxicity of the various extracts was assessed and the essential oil extracts exhibited superior activity. The phenolic composition and biological activity of the fruits indicate that their consumption is beneficial to health and also that their incorporation into functional foods and nutraceuticals should be considered.Keywords: phytochemicals, antimicrobial, cytotoxic, Rubus rosifolius
Procedia PDF Downloads 397180 DLtrace: Toward Understanding and Testing Deep Learning Information Flow in Deep Learning-Based Android Apps
Authors: Jie Zhang, Qianyu Guo, Tieyi Zhang, Zhiyong Feng, Xiaohong Li
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With the widespread popularity of mobile devices and the development of artificial intelligence (AI), deep learning (DL) has been extensively applied in Android apps. Compared with traditional Android apps (traditional apps), deep learning based Android apps (DL-based apps) need to use more third-party application programming interfaces (APIs) to complete complex DL inference tasks. However, existing methods (e.g., FlowDroid) for detecting sensitive information leakage in Android apps cannot be directly used to detect DL-based apps as they are difficult to detect third-party APIs. To solve this problem, we design DLtrace; a new static information flow analysis tool that can effectively recognize third-party APIs. With our proposed trace and detection algorithms, DLtrace can also efficiently detect privacy leaks caused by sensitive APIs in DL-based apps. Moreover, using DLtrace, we summarize the non-sequential characteristics of DL inference tasks in DL-based apps and the specific functionalities provided by DL models for such apps. We propose two formal definitions to deal with the common polymorphism and anonymous inner-class problems in the Android static analyzer. We conducted an empirical assessment with DLtrace on 208 popular DL-based apps in the wild and found that 26.0% of the apps suffered from sensitive information leakage. Furthermore, DLtrace has a more robust performance than FlowDroid in detecting and identifying third-party APIs. The experimental results demonstrate that DLtrace expands FlowDroid in understanding DL-based apps and detecting security issues therein.Keywords: mobile computing, deep learning apps, sensitive information, static analysis
Procedia PDF Downloads 180179 Chaupadi Practice: A Cruel Discrimination against Women a Case Study of Achham District of Nepal
Authors: Santosh Thapa, Sankar Gurung
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Chaupadi is a tradition widely practiced in the far and mid-western region of Nepal. It is a practice where girls and women are not allowed to inter the house and touch the food, water and milk during their menstruation period of 4-7 days. They have to spend all the nights during the period in a specific hut (Chhaupadi Goth) which is a bit far from their residence where they faces various kinds of risk and violence like bullying, snakes and insect bite, wild animal attack etc. Sometimes the girls even do not go to school during their menstruation periods. After childbirth, the woman must stay in a cow shed for 11 days in such Chhaupadi practiced areas. This study limits the Achham district of the far western region which is the most vulnerable Chhaupadi practicing district. Several governmental and non-governmental organizations have been involving and spending huge amount of money for capacity building and awareness raising campaign for last 2 decades but still 9 out of 75 Village Development Committees (VDCs) have been partially practicing Chaupadi in the district. This study shows that the school attendance rate of the girls during the period have visibly increased which helps to increase the number of the girl graduation as well. Similarly, the practice of Chhaupadi is one of the reasons for increasing the number of cases of uterus prolapsus and poor reproductive health of women and girls. Triggering tools are the one of the best ways to accelerate the awareness campaign in the VDCs. This study recommends that the local bodies should coordinate and lead the overall awareness campaign program to sustain the Chaupadi free VDCs.Keywords: awareness campaign, chaupadi practice, gender discrimination, violence
Procedia PDF Downloads 312178 Mayan Culture and Attitudes towards Sustainability
Authors: Sarah Ryu
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Agricultural methods and ecological approaches employed by the pre-colonial Mayans may provide valuable insights into forest management and viable alternatives for resource sustainability in the face of major deforestation across Central and South America.Using a combination of observation data collected from the modern indigenous inhabitants near Mixco in Guatemala and historical data, this study was able to create a holistic picture of how the Maya maintained their ecosystems. Surveys and observations were conducted in the field, over a period of twelve weeks across two years. Geographic and archaeological data for this area was provided by Guatemalan organizations such as the Universidad de San Carlos de Guatemala. Observations of current indigenous populations around Mixco showed that they adhered to traditional Mayan methods of agriculture, such as terrace construction and arboriculture. Rather than planting one cash crop as was done by the Spanish, indigenous peoples practice agroforestry, cultivating forests that would provide trees for construction material, wild plant foods, habitat for game, and medicinal herbs. The emphasis on biodiversity prevented deforestation and created a sustainable balance between human consumption and forest regrowth. Historical data provided by MayaSim showed that the Mayans successfully maintained their ecosystems from about 800BCE to 700CE. When the Mayans practiced natural resource conservation and cultivated a harmonious relationship with the forest around them, they were able to thrive and prosper alongside nature. Having lasted over a thousand years, the Mayan empire provides a valuable lesson in sustainability and human attitudes towards the environment.Keywords: biodiversity, forestry, mayan, sustainability
Procedia PDF Downloads 177177 Prediction of Solanum Lycopersicum Genome Encoded microRNAs Targeting Tomato Spotted Wilt Virus
Authors: Muhammad Shahzad Iqbal, Zobia Sarwar, Salah-ud-Din
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Tomato spotted wilt virus (TSWV) belongs to the genus Tospoviruses (family Bunyaviridae). It is one of the most devastating pathogens of tomato (Solanum Lycopersicum) and heavily damages the crop yield each year around the globe. In this study, we retrieved 329 mature miRNA sequences from two microRNA databases (miRBase and miRSoldb) and checked the putative target sites in the downloaded-genome sequence of TSWV. A consensus of three miRNA target prediction tools (RNA22, miRanda and psRNATarget) was used to screen the false-positive microRNAs targeting sites in the TSWV genome. These tools calculated different target sites by calculating minimum free energy (mfe), site-complementarity, minimum folding energy and other microRNA-mRNA binding factors. R language was used to plot the predicted target-site data. All the genes having possible target sites for different miRNAs were screened by building a consensus table. Out of these 329 mature miRNAs predicted by three algorithms, only eight miRNAs met all the criteria/threshold specifications. MC-Fold and MC-Sym were used to predict three-dimensional structures of miRNAs and further analyzed in USCF chimera to visualize the structural and conformational changes before and after microRNA-mRNA interactions. The results of the current study show that the predicted eight miRNAs could further be evaluated by in vitro experiments to develop TSWV-resistant transgenic tomato plants in the future.Keywords: tomato spotted wild virus (TSWV), Solanum lycopersicum, plant virus, miRNAs, microRNA target prediction, mRNA
Procedia PDF Downloads 155176 An Exploration of the Place of Buddhism in the Tham Luang Cave Rescue and Its Aftermath
Authors: Hamish de Nett
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On 23rd June 2018, twelve young footballers from the Wild Boar Academy and their coach went to explore the Tham Luang cave in the Doi Nang Non mountain range in Chiang Rai Province, Northern Thailand. Whilst they were inside the cave, monsoon rains hit, and the complex became partially flooded. In the following days, Thai Navy SEALs and an international team of expert divers assembled at the cave complex in order to rescue the boys. Although it was only marginally reported in the Western press, Buddhism and ritual activities played a major role in the rescue and its aftermath. This paper utilises numerous news articles and books written by reporters who covered the cave rescue to uncover what the place of Buddhism was in the Tham Luang cave rescue. This paper initially sets out the development of Thai Buddhism and the Thai nation state, paying particular note to the tension in Thai Buddhism between Buddhism as it is popularly practised and normative, state-favoured Buddhism. Secondly, this paper demonstrates that, during the Tham Luang cave rescue, Buddhism helped people cope with the disaster, provided an explanation for its occurrence, and allowed bystanders some efficacy in the process. Thirdly, this paper discusses how Buddhism helped people to give thanks after the rescue, achieve reconciliation, and gain closure. Finally, this paper analyses how the government and the political sphere utilised Buddhism during the rescue. The conclusion reached is that the Buddhism practiced during the Tham Luang cave rescue and its aftermath is representative of the wider tension between popular Buddhism and normative state-favoured Buddhism that is currently present within Thai Buddhism and has been for centuries.Keywords: cave rescue, contemporary Buddhism, lived religion, Thai Buddhism, Tham Luang cave rescue
Procedia PDF Downloads 130175 Antioxidant and Antimicrobial Activities of Matricaria pubscens Extracts: A Wild Space of North African Pharmacopeia
Authors: Abdelouahab Dehimati, Fatiha Bedjou
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This study focused on the antioxidant and antimicrobial activity of four extracts from the plant Matricaria pubscens (Asteraceae) harvest in the region of Ghardaia, the northern Sahara of Algeria. The different extracts were analyzed for their content of phenolic compounds and their biological activities. The ethanol extract expresses a better extraction yield (44.22%). We have first performed the quantitative colorimetric methods for total polyphenols. Wherein the aqueous extract shows the highest total polyphenol content and total flavonoid (216.66±2.58 mg Eq GA/g and 111.04±0.49 mg Eq Q/g E, respectively) and ethanol extract 50% total tannins content (68.88±2.72 mg Eq AT/g E). The evaluation of the antioxidant activity of extracts of Matricaria pubscens by the arbitrary value IC50. The ethanol 50% extract is expressed strong activity with an IC50 14.19±1.25 mg/m against the DPPH radical and 11.66±0.53 mg/ml against the ABTS radical). In addition, the aqueous extract showed strong reducing power with an IC50 (48.61±1.14 mg/ml). However, the results obtained by the reducing power of phosphomolybdat the test are calculated by the iron maximum absorbance where ethanol extract 50% gives an absorbance of about 1.641 ± 0.01nm. Otherwise, methanol 70% and butanol 80% extracts gave a very large chelating effect of iron with an IC50 (38.38±0.01 μg/ml and 38.58±0.04 μg/ml respectively). By the method of disc Diffuson, the results of the antimicrobial activity are achieved butanolic extract 80% shows high activity towards MRSA (MIC: 3.51mg/ml; BMC>100 mg/ml). Their shares, the extracts were the most active for the antifungal test, the butanol 80% extract was the most active against A. niger (MIC: 12.5 mg/ml; FMC>100 mg/ml). These preliminary results could be used to justify the traditional use of this plant and their phenolic compounds could be exploited for therapeutic purposes, such as antioxidants and antimicrobial effects.Keywords: Matricaria pubscens, phenolic compounds, antioxidant activity, antimicrobial activity, IC50, MIC
Procedia PDF Downloads 276174 Water Treatment Using Eichhornia crassipes and Avifauna Control in The "La Mansión" Pond
Authors: Milda A. Cruz-Huaranga, Natalí Carbo-Bustinza, Javier Linkolk López-Gonzales, K. Depaz, Gina M. Tito T., Soledad Torres-Calderón
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The objective of this study was to improve water quality in the “La Mansión” pond in order to irrigate green spaces on the Peruvian Union University campus (Lima, Peru) using the aquatic species Eichhornia Crassipes. Furthermore, tree trimming and cleaning activities were performed that reduced water pollution caused by organic deposits and feathers from wild birds. The impaired waterbody is located on the campus of the Peruvian Union University, 580 meters above sea level, with a volume of 6,405.336 m3, an area of 3,050.16 m2, 256.81 m perimeter, and 0.12 m3/s input flow. Seven 1.8 m2 floating systems were implemented, with 12 common water hyacinth plants in each system. Before implementing this system, a water quality analysis was performed to analyse the physical-chemical, microbiological, and organoleptic parameters. The pre-analysis revealed the pond’s critical condition, with electrical conductivity: 556 mg/l; phosphate: < 0.5; pH: 7.06; total solids: 412 mg/l; arsenic: <0.01; lead: 0.115; BOD5: 14; COD: 16.94; dissolved oxygen: 13; total coliforms: 24000 MCL/100 ml; and thermo-tolerant coliforms: 11000 MCL/100 ml. After implementing the system, the following results were obtained: EC: 495 mg/l; DO:9.2 mg/l; TS: 235 mg/l; BOD5: 7.7; COD: 8.47; Pb: 0.001 mg/l; TC: 460 MCL/100 ml; FC: 240 MCL/100 ml. Thus, we confirmed that the system is 78.79% efficient regarding the Peruvian ECA (Environmental Quality Standards) established for water according to DS #015-2015-MINAM. Therefore, the water is suitable for plant irrigation. Finally, we concluded that treating wastewater with the species Eichhornia Crassipes is efficient since an improvement was achieved in the impaired waterbody.Keywords: Eichhornia crassipes, plantlets, cleaning, impaired waterbody, pond
Procedia PDF Downloads 141173 Implications of Dehusking and Aqueous Soaking on Anti-nutrients, Phytochemical Screening and Antioxidants Properties of Jack Beans (Canavalia Ensiformis L. DC)
Authors: Oseni Margaret Oladunni, Ogundele Joan Olayinka, Olusanya Olalekan Samuel, Akinniyi Modupe Olakintan
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The world's growing population is pushing humans to look for alternative food sources among underutilised or wild plants. One of these food sources has been identified as Canavalia enisiformis, or jack beans. The only issue with using jack beans is that they contain anti-nutrient chemicals, which must be removed or diminished in order for them to be fit for human consumption. The objective of this study is to determine the nutritional and industrial utility of Canavalia enisiformis by analysing the anti-nutrient, phytochemical, and antioxidant composition of raw whole seed and soaking dehusked seeds using established procedures. Phytate (23.48±0.24, 15.24±0.41 and 14.83±0.00), oxalate (4.32±0.09, 3.96±0.09 and 2.88±0.09), tannins (22.77±0.73, 18.68±0.03 and 17.50±0.46), and lectins (6.67±0.04, 6.20±0.01 and 6.42±0.07) exhibited the highest anti-nutrient values in raw whole seed and, at the very least, in dehusked, soaked seeds. The samples were subjected to phytochemical screening, which detected the presence of cardiac glycosides as well as anthraquinones, alkaloids, tannins, saponins, steroids, flavonoids, terpenoids, phlobatannins, and flavonoids. Due to the reduction in phytochemical contents quantified as a result of dehusking and soaking, phenolbatannins and anthraquinones were not found in the samples. The research findings also demonstrated elevated concentrations of several plausible phytochemical components with potential medical value, with the raw whole seed exhibiting the greatest capacity to scavenge free radicals. Accordingly, the study's findings validate the seed's therapeutic applications and imply that it might be an inexpensive source of antioxidants for humans and animals alike.Keywords: dehusking, soaking, anti-nutrients, antioxidants, jack bean
Procedia PDF Downloads 50172 PPRA Regulates DNA Replication Initiation and Cell Morphology in Escherichia coli
Authors: Ganesh K. Maurya, Reema Chaudhary, Neha Pandey, Hari S. Misra
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PprA, a pleiotropic protein participating in radioresistance, has been reported for its roles in DNA replication initiation, genome segregation, cell division and DNA repair in polyextremophile Deinococcus radiodurans. Interestingly, expression of deinococcal PprA in E. coli suppresses its growth by reducing the number of colony forming units and provides better resistance against γ-radiation than control. We employed different biochemical and cell biology studies using PprA and its DNA binding/polymerization mutants (K133E & W183R) in E. coli. Cells expressing wild type PprA or its K133E mutant showed reduction in the amount of genomic DNA as well as chromosome copy number in comparison to W183R mutant of PprA and control cells, which suggests the role of PprA protein in regulation of DNA replication initiation in E. coli. Further, E. coli cells expressing PprA or its mutants exhibited different impact on cell morphology than control. Expression of PprA or K133E mutant displayed a significant increase in cell length upto 5 folds while W183R mutant showed cell length similar to uninduced control cells. We checked the interaction of deinococcal PprA and its mutants with E. coli DnaA using Bacterial two-hybrid system and co-immunoprecipitation. We observed a functional interaction of EcDnaA with PprA and K133E mutant but not with W183R mutant of PprA. Further, PprA or K133E mutant has suppressed the ATPase activity of EcDnaA but W183R mutant of PprA failed to do so. These observations suggested that PprA protein regulates DNA replication initiation and cell morphology of surrogate E. coli.Keywords: DNA replication, radioresistance, protein-protein interaction, cell morphology, ATPase activity
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