Search results for: nucleic acid extraction
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 5052

Search results for: nucleic acid extraction

942 The Comparison Study of Human Microbiome in Chronic Rhinosinusitis between Adults and Children

Authors: Il Ho Park, Joong Seob Lee, Sung Hun Kang, Jae-Min Shin, Il Seok Park, Seok Min Hong, Seok Jin Hong

Abstract:

Introduction: The human microbiota is the aggregate of microorganisms, and the bacterial microbiome of the human digestive tract contributes to both health and disease. In health, bacteria are key components in the development of mucosal barrier function and in innate and adaptive immune responses, and they also work to suppress the establishment of pathogens. In human upper airway, the sinonasal microbiota might play an important role in chronic rhinosinusitis (CRS). The purpose of this study is to investigate the human upper airway microbiome in CRS patients and to compare the sinonasal microbiome of adults with children. Materials and methods: A total of 19 samples from 19 patients (Group1; 9 CRS in children, aged 5 to 14 years versus Group 2; 10 CRS in adults aged 21 to 59 years) were examined. Swabs were collected from the middle meatus and/or anterior ethmoid region under general anesthesia during endoscopic sinus surgery or tonsillectomy. After DNA extraction from swab samples, we analysed bacterial microbiome consortia using 16s rRNA gene sequencing approach (the Illumina MiSeq platform). Results: In this study, relatively abundance of the six bacterial phyla and tremendous genus and species found in substantial amounts in the individual sinus swab samples, include Corynebacterium, Hemophilus, Moraxella, and Streptococcus species. Anaerobes like Fusobacterium and Bacteroides were abundantly present in the children group, Bacteroides and Propionibacterium were present in adults group. In genus, Haemophilus was the most common CRS microbiome in children and Corynebacterium was the most common CRS microbiome in adults. Conclusions: Our results show the diversity of human upper airway microbiome, and the findings will suggest that CRS is a polymicrobial infection. The Corynebacterium and Hemophilus may live as commensals on mucosal surfaces of sinus in the upper respiratory tract. The further study will be needed for analysis of microbiome-human interactions in upper airway and CRS.

Keywords: microbiome, upper airway, chronic rhinosinusitis, adult and children

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941 Dietary N-6/N-3 PUFA Ratios Affect the Homeostasis of CD4+ T Cells in Mice with Dextran Sulfate Sodium-Induced Colitis

Authors: Cyoung-Huei Huang, Chiu-Li Yeh, Man-Hui Pai, Sung-Ling Yeh

Abstract:

This study evaluated the effect of different dietary n-6/n-3 polyunsaturated fatty acid (PUFA) ratios on modulating helper T (Th) and regulatory T (Treg) lymphocytes in mice with dextran sulfate sodium (DSS)-induced colitis. There were 3 control and 3 colitis groups in this study. Mice were fed for 24 d with an AIN-93G diet either with soybean oil (S), a mixture of soybean oil and low fish oil content (LF) or high fish oil content (HF). The ratio of n-6/n-3 PUFA in the LF diet was 4:1, and that in the HF diet was 2:1. The control groups drank distilled water while colitis groups provided 2% DSS in drinking water during day 15-19. All mice drank distilled water from day 20-24 for recovery and sacrificed on day 25. The results showed that colitis resulted in higher Th1, Th2, and Th17 and lower Treg percentages in the blood. Also, plasma haptoglobin and proinflammatory chemokines were elevated in colon lavage fluid. Colitic groups with fish oil had lower inflammatory mediators in the plasma and colon lavage fluid. Further, the percentages of Th1, Th2, and Th17 cells in the blood were lower, whereas Treg cell percentages were higher than those in the soybean oil group. The colitis group with n-6/n-3 PUFA ratio 2:1 had more pronounce effects than ratio 4:1. These results suggest that diets with an n-6/n-3 PUFA ratio of 2:1 or 4:1 regulate the Th/Treg balance and attenuate inflammatory mediator production in colitis. Compared to the n-6/n-3 PUFA ratio 4:1, the ratio of 2:1 was more effective in reducing inflammatory reactions in DSS-induced colitis.

Keywords: inflammatory bowel disease, n-3 polyunsaturated fatty acids, helper T lymphocyte, regulatory T lymphocyte

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940 Contribution of NLRP3 Inflammasome to the Protective Effect of 5,14-HEDGE, A 20-HETE Mimetic, against LPS-Induced Septic Shock in Rats

Authors: Bahar Tunctan, Sefika Pinar Kucukkavruk, Meryem Temiz-Resitoglu, Demet Sinem Guden, Ayse Nihal Sari, Seyhan Sahan-Firat, Mahesh P. Paudyal, John R. Falck, Kafait U. Malik

Abstract:

We hypothesized that 20-hydroxyeicosatetraenoic acid (20-HETE) mimetics such as N-(20-hydroxyeicosa-5[Z],14[Z]-dienoyl)glycine (5,14-HEDGE) may be beneficial for preventing mortality due to inflammation induced by lipopolysaccharide (LPS). This study aims to assess the effect of 5,14-HEDGE on the LPS-induced changes in nucleotide binding domain and leucine-rich repeat protein 3 (NLRP3)/apoptosis-associated speck-like protein containing a caspase activation and recruitment domain (ASC)/pro-caspase-1 inflammasome. Rats were injected with saline (4 ml/kg) or LPS (10 mg/kg) at time 0. Blood pressure and heart rate were measured using a tail-cuff device. 5,14-HEDGE (30 mg/kg) was administered to rats 1 h after injection of saline or LPS. The rats were sacrificed 4 h after saline or LPS injection and kidney, heart, thoracic aorta, and superior mesenteric artery were isolated for measurement of caspase-1/11 p20, NLRP3, ASC, and β-actin proteins as well as interleukin-1β (IL-1β) levels. Blood pressure decreased by 33 mmHg and heart rate increased by 63 bpm in the LPS-treated rats. In the LPS-treated rats, tissue protein expression of caspase-1/11 p20, NLRP3, and ASC in addition to IL-1β levels were increased. 5,14-HEDGE prevented the LPS-induced changes. Our findings suggest that inhibition of renal, cardiac, and vascular formation/activity of NLRP3/ASC/pro-caspase-1 inflammasome involved in the protective effect of 5,14-HEDGE on LPS-induced septic shock in rats. This work was financially supported by the Mersin University (2015-AP3-1343) and USPHS NIH (PO1 HL034300).

Keywords: 5, 14-HEDGE, lipopolysaccharide, NLRP3, inflammasome, septic shock

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939 Ultrasonic Treatment of Baker’s Yeast Effluent

Authors: Emine Yılmaz, Serap Fındık

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Baker’s yeast industry uses molasses as a raw material. Molasses is end product of sugar industry. Wastewater from molasses processing presents large amount of coloured substances that give dark brown color and high organic load to the effluents. The main coloured compounds are known as melanoidins. Melanoidins are product of Maillard reaction between amino acid and carbonyl groups in molasses. Dark colour prevents sunlight penetration and reduces photosynthetic activity and dissolved oxygen level of surface waters. Various methods like biological processes (aerobic and anaerobic), ozonation, wet air oxidation, coagulation/flocculation are used to treatment of baker’s yeast effluent. Before effluent is discharged adequate treatment is imperative. In addition to this, increasingly stringent environmental regulations are forcing distilleries to improve existing treatment and also to find alternative methods of effluent management or combination of treatment methods. Sonochemical oxidation is one of the alternative methods. Sonochemical oxidation employs ultrasound resulting in cavitation phenomena. In this study, decolorization of baker’s yeast effluent was investigated by using ultrasound. Baker’s yeast effluent was supplied from a factory which is located in the north of Turkey. An ultrasonic homogenizator used for this study. Its operating frequency is 20 kHz. TiO2-ZnO catalyst has been used as sonocatalyst. The effects of molar proportion of TiO2-ZnO, calcination temperature and time, catalyst amount were investigated on the decolorization of baker’s yeast effluent. The results showed that prepared composite TiO2-ZnO with 4:1 molar proportion treated at 700°C for 90 min provides better result. Initial decolorization rate at 15 min is 3% without catalyst, 14,5% with catalyst treated at 700°C for 90 min respectively.

Keywords: baker’s yeast effluent, decolorization, sonocatalyst, ultrasound

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938 Chrysin-Loaded PLGA-PEG Nanoparticles Designed for Enhanced Inhibitory Effect on the Breast Cancer Cell Line

Authors: Faraz Zarghami, Elham Anari, Nosratollah Zarghami, Yones Pilehvar-Soltanahmadi, Abolfazl Akbarzadeh, Sepideh Jalilzadeh-Tabrizi

Abstract:

The development of nanotherapy has presented a new method of drug delivery targeted directly to the neoplasmic tissues, to maximize the action with fewer dose requirements. In the past two decades, poly(lactic-co-glycolic acid) (PLGA) has frequently been investigated by many researchers and is a popular polymeric candidate, due to its biocompatibility and biodegradability, exhibition of a wide range of erosion times, tunable mechanical properties, and most notably, because it is a FDA-approved polymer. Chrysin is a natural flavonoid which has been reported to have some significant biological effects on the processes of chemical defense, nitrogen fixation, inflammation, and oxidation. However, the low solubility in water decreases its bioavailability and consequently disrupts the biomedical benefits. Being loaded with PLGA-PEG increases chrysin solubility and drug tolerance, and decreases the discordant effects of the drug. The well-structured chrysin efficiently accumulates in the breast cancer cell line (T47D). In the present study, the structure and chrysin loading were delineated using proton nuclear magnetic resonance (HNMR), Fourier-transform infrared spectroscopy (FT-IR), and scanning electron microscopy (SEM), and the in vitro cytotoxicity of pure and nanochrysin was studied by the MTT assay. Next, the RNA was exploited and the cytotoxic effects of chrysin were studied by real-time PCR. In conclusion, the nanochrysin therapy developed is a novel method that could increase cytotoxicity to cancer cells without damaging the normal cells, and would be promising in breast cancer therapy.

Keywords: MTT assay, chrysin, flavonoids, nanotherapy

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937 Spontaneous Eruption of Impacted Teeth While Awaiting Surgical Intervention

Authors: Alison Ryan, Himani Chhabra, Mohammed Dungarwalla, Judith Jones

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Background: Impacted and ectopic teeth present in 1-2% of orthodontic patients and often require joint surgical and orthodontic management. The authors present two patients undergoing orthodontic treatment, where the impacted teeth, in a hopeless position, spontaneously erupted during the period of cessation of general anaesthetic lists during the COVID-19 pandemic. Patient information: A healthy 11-year-old boy was referred to the Department of Oral and Maxillofacial Surgery for the management of a mesioangular impacted LR7. The patient was seen by the joint oral surgery/orthodontic team, who planned for the removal of the LR7 under general anaesthetic. A healthy 13-year-old boy was referred to the same Department and team for surgical extraction of unerupted and buccally impacted UL3 and UR3 under general anaesthetic. Management and outcome: The majority of elective dental-alveolar work ceased as a result of the global pandemic. On resumption of activity, the first patient was reviewed in July 2021. The LR7 had spontaneously erupted in a favourable position, and following MDT review, a decision was made to forgo any further surgical intervention. The second patient was reviewed in July 2021. The UL3 had clinically erupted, and there was radiographic evidence of favourable movement of UR3. Due to the nature of the patient’s malocclusion, the decision was made to proceed with the extractions as previously planned. Key Learning Points: Severely impacted teeth do have a prospect of spontaneous eruption or alignment without clinical intervention, and current literature states the initial location, axial inclination, degree of root formation, and relation of the impacted tooth to adjacent teeth roots may influence spontaneous eruption. There is potential to introduce a period of observation to account for this possibility in the developing dentition, with the aim of reducing the unnecessary need for surgical intervention. This could help prevent episodes of general anaesthetic and allocate theatre space more appropriately.

Keywords: spontaneous eruption, impaction, observation, hopeless position, surgical, orthodontic, change in treatment plan

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936 Cerebrum Maturity Damage Induced by Fluoride in Suckling Mice

Authors: Hanen Bouaziz, Françoise Croute, Najiba Zeghal

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In order to investigate the toxic effects of fluoride on cerebrum maturity of suckling mice, we treated adult female mice of Swiss Albinos strain by 500 ppm NaF in their drinking water from the 15th day of pregnancy until the day 14 after delivery. All mice were sacrificed on day 14 after parturition. During treatment, levels of thiobarbituric acid reactive substances, the marker of lipid peroxidation extend, increased, while the activities of the antioxidant enzymes such as glutathione peroxidase, superoxide dismutase and catalase and the level of glutathione decreased significantly in cerebellum compared with those of the control group. These results suggested that fluoride enhanced oxidative stress, thereby disturbing the antioxidant defense of nursing pups. In addition, acetylcholinesterase activity in cerebellum was inhibited after treatment with fluoride. In cerebellum of mice, migration of neurons from the external granular layer to the internal granular layer occurred postnatally. Key guidance signals to these migrating neurons were provided by laminin, an extracellular matrix protein fixed to the surface of astrocytes. In the present study, we examined the expression and distribution of laminin in cerebellum of 14-day-old mice. Immunoreactive laminin was disappeared by postnatal day 14 in cerebellum parenchyma of control pups and was restricted to vasculature despite the continued presence of granular cells in the external granular layer. In contrast, in cerebellum of NaF treated pups, laminin was deposited in organised punctuate clusters in the molecular layer. These data indicated that the disruption of laminin distribution might play a major role in the profound derangement of neuronal migration observed in cerebellum of NaF treated pups.

Keywords: acetylcholinesterase activity, cerebellum, laminin, oxidative stress, suckling mice

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935 Biochemical and Pomological Variability among 14 Moroccan and Foreign Cultivars of Prunus dulcis

Authors: H. Hanine, H. H'ssaini, M. Ibno Alaoui, A. Nablousi, H. Zahir, S. Ennahli, H. Latrache, H. Zine Abidine

Abstract:

Biochemical and pomological variability among 14 cultivars of Prunus dulcis planted in a germoplasm collection site in Morocco were evaluated. Almond samples from six local and eight foreign cultivars (France, Italy, Spain, and USA) were characterized. Biochemical and pomological data revealed significant genetic variability among the 14 cultivars; local cultivars exhibited higher total polyphenol content. Oil content ranged from 35 to 57% among cultivars; both Texas and Toundout genotypes recorded the highest oil content. Total protein concentration from select cultivars ranged from 50 mg/g in Ferraduel to 105 mg/g in Rizlane1 cultivars. Antioxidant activity of almond samples was examined by a DPPH (1,1-diphenyl-2-picrylhydrazyl) radical-scavenging assay; the antioxidant activity varied significantly within the cultivars, with IC50 (the half-maximal inhibitory concentration) values ranging from 2.25 to 20 mg/ml. Autochthonous cultivars originated from the Oujda region exhibited higher tegument total polyphenol and amino acid content compared to others. The genotype Rizlane2 recorded the highest flavonoid content. Pomological traits revealed a large variability within the almond germplasms. The hierarchical clustering analysis of all the data regarding pomological traits distinguished two groups with some particular genotypes as distinct cultivars, and groups of cultivars as polyclone varieties. These results strongly exhibit a potential use of Moroccan-originated almonds as potential clones for future selection due to their nutritional values and pomological traits compared to well-established cultivars.

Keywords: antioxidant activity, DDPH, Moroccan almonds, Prunus dulcis

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934 Unsupervised Part-of-Speech Tagging for Amharic Using K-Means Clustering

Authors: Zelalem Fantahun

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Part-of-speech tagging is the process of assigning a part-of-speech or other lexical class marker to each word into naturally occurring text. Part-of-speech tagging is the most fundamental and basic task almost in all natural language processing. In natural language processing, the problem of providing large amount of manually annotated data is a knowledge acquisition bottleneck. Since, Amharic is one of under-resourced language, the availability of tagged corpus is the bottleneck problem for natural language processing especially for POS tagging. A promising direction to tackle this problem is to provide a system that does not require manually tagged data. In unsupervised learning, the learner is not provided with classifications. Unsupervised algorithms seek out similarity between pieces of data in order to determine whether they can be characterized as forming a group. This paper explicates the development of unsupervised part-of-speech tagger using K-Means clustering for Amharic language since large amount of data is produced in day-to-day activities. In the development of the tagger, the following procedures are followed. First, the unlabeled data (raw text) is divided into 10 folds and tokenization phase takes place; at this level, the raw text is chunked at sentence level and then into words. The second phase is feature extraction which includes word frequency, syntactic and morphological features of a word. The third phase is clustering. Among different clustering algorithms, K-means is selected and implemented in this study that brings group of similar words together. The fourth phase is mapping, which deals with looking at each cluster carefully and the most common tag is assigned to a group. This study finds out two features that are capable of distinguishing one part-of-speech from others these are morphological feature and positional information and show that it is possible to use unsupervised learning for Amharic POS tagging. In order to increase performance of the unsupervised part-of-speech tagger, there is a need to incorporate other features that are not included in this study, such as semantic related information. Finally, based on experimental result, the performance of the system achieves a maximum of 81% accuracy.

Keywords: POS tagging, Amharic, unsupervised learning, k-means

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933 Stabilization of Pb, Cr, Cd, Cu and Zn in Solid Waste and Sludge Pyrolysis by Modified Vermiculite

Authors: Yuxuan Yang, Zhaoping Zhong

Abstract:

Municipal solid waste and sludge are important sources of waste energy and their proper disposal is of great importance. Pyrolysis can fully decompose solid wastes and sludge, and the pyrolysis products (charcoal, oil and gas) have important recovery values. Due to the complex composition of solid wastes and sludge, the pyrolysis process at high temperatures is prone to heavy metal emissions, which are harmful to humans and the environment and reduce the safety of pyrolysis products. In this paper, heavy metal emissions during pyrolysis of municipal sewage sludge, paper mill sludge, municipal domestic waste, and aged refuse at 450-650°C were investigated and the emissions and hazards of heavy metals (Pb, Cr, Cd, Cu and Zn) were effectively reduced by adding modified vermiculite as an additive. The vermiculite was modified by intercalation with cetyltrimethylammonium bromide, which resulted in more than twice the original layer spacing of the vermiculite. Afterward, the interpolated vermiculite was made into vermiculite flakes by exfoliation modification. After that, the expansion rate of vermiculite flakes was increased by Mg2+ modification and thermal activation. The expanded vermiculite flakes were acidified to improve the textural characteristics of the vermiculite. The modified vermiculite was analysed by XRD, FT-IR, BET and SEM to clarify the modification effect. The incorporation of modified vermiculite resulted in more than 80% retention of all heavy metals at 450°C. Cr, Cu and Zn were better retained than Pb and Cd. The incorporation of modified vermiculite effectively reduced the risk of heavy metals, and all risks were low for Pb, Cr, Cu and Zn. The toxicity of all heavy metals was greatly reduced by the incorporation of modified vermiculite and the morphology of heavy metals was transformed from Exchangeable and acid-soluble (F1) and Reducible (F2) to Oxidizable (F3) and Residual (F4). In addition, the increase in temperature favored the stabilization of heavy metal forms. This study provides a new insight into the cleaner use of energy and the safe management of solid waste.

Keywords: heavy metal, pyrolysis, vermiculite, solid waste

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932 Development of New Localized Surface Plasmon Resonance Interfaces Based on ITO Au NPs/ Polymer for Nickel Detection

Authors: F. Z. Tighilt, N. Belhaneche-Bensemra, S. Belhousse, S. Sam, K. Lasmi, N. Gabouze

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Recently, the gold nanoparticles (Au NPs) became an active multidisciplinary research topic. First, Au thin films fabricated by alkylthiol-functionalized Au NPs were found to have vapor sensitive conductivities, they were hence widely investigated as electrical chemiresistors for sensing different vapor analytes and even organic molecules in aqueous solutions. Second, Au thin films were demonstrated to have speciallocalized surface plasmon resonances (LSPR), so that highly ordered 2D Au superlattices showed strong collective LSPR bands due to the near-field coupling of adjacent nanoparticles and were employed to detect biomolecular binding. Particularly when alkylthiol ligands were replaced by thiol-terminated polymers, the resulting polymer-modified Au NPs could be readily assembled into 2D nanostructures on solid substrates. Monolayers of polystyrene-coated Au NPs showed typical dipolar near-field interparticle plasmon coupling of LSPR. Such polymer-modified Au nanoparticle films have an advantage that the polymer thickness can be feasibly controlled by changing the polymer molecular weight. In this article, the effect of tin-doped indium oxide (ITO) coatings on the plasmonic properties of ITO interfaces modified with gold nanostructures (Au NSs) is investigated. The interest in developing ITO overlayers is multiple. The presence of a con-ducting ITO overlayer creates a LSPR-active interface, which can serve simultaneously as a working electrode in an electro-chemical setup. The surface of ITO/ Au NPs contains hydroxyl groups that can be used to link functional groups to the interface. Here the covalent linking of nickel /Au NSs/ITO hybrid LSPR platforms will be presented.

Keywords: conducting polymer, metal nanoparticles (NPs), LSPR, poly (3-(pyrrolyl)–carboxylic acid), polypyrrole

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931 Numerical Study of Natural Convection in Isothermal Open Cavities

Authors: Gaurav Prabhudesai, Gaetan Brill

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The sun's energy source comes from a hydrogen-to-helium thermonuclear reaction, generating a temperature of about 5760 K on its outer layer. On account of this high temperature, energy is radiated by the sun, a part of which reaches the earth. This sunlight, even after losing part of its energy en-route to scattering and absorption, provides a time and space averaged solar flux of 174.7 W/m^2 striking the earth’s surface. According to one study, the solar energy striking earth’s surface in one and a half hour is more than the energy consumption that was recorded in the year 2001 from all sources combined. Thus, technology for extraction of solar energy holds much promise for solving energy crisis. Of the many technologies developed in this regard, Concentrating Solar Power (CSP) plants with central solar tower and receiver system are very impressive because of their capability to provide a renewable energy that can be stored in the form of heat. One design of central receiver towers is an open cavity where sunlight is concentrated into by using mirrors (also called heliostats). This concentrated solar flux produces high temperature inside the cavity which can be utilized in an energy conversion process. The amount of energy captured is reduced by losses occurring at the cavity through all three modes viz., radiation to the atmosphere, conduction to the adjoining structure and convection. This study investigates the natural convection losses to the environment from the receiver. Computational fluid dynamics were used to simulate the fluid flow and heat transfer of the receiver; since no analytical solution can be obtained and no empirical correlations exist for the given geometry. The results provide guide lines for predicting natural convection losses for hexagonal and circular shaped open cavities. Additionally, correlations are given for various inclination angles and aspect ratios. These results provide methods to minimize natural convection through careful design of receiver geometry and modification of the inclination angle, and aspect ratio of the cavity.

Keywords: concentrated solar power (CSP), central receivers, natural convection, CFD, open cavities

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930 Hydraulic Characteristics of Mine Tailings by Metaheuristics Approach

Authors: Akhila Vasudev, Himanshu Kaushik, Tadikonda Venkata Bharat

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A large number of mine tailings are produced every year as part of the extraction process of phosphates, gold, copper, and other materials. Mine tailings are high in water content and have very slow dewatering behavior. The efficient design of tailings dam and economical disposal of these slurries requires the knowledge of tailings consolidation behavior. The large-strain consolidation theory closely predicts the self-weight consolidation of these slurries as the theory considers the conservation of mass and momentum conservation and considers the hydraulic conductivity as a function of void ratio. Classical laboratory techniques, such as settling column test, seepage consolidation test, etc., are expensive and time-consuming for the estimation of hydraulic conductivity variation with void ratio. Inverse estimation of the constitutive relationships from the measured settlement versus time curves is explored. In this work, inverse analysis based on metaheuristics techniques will be explored for predicting the hydraulic conductivity parameters for mine tailings from the base excess pore water pressure dissipation curve and the initial conditions of the mine tailings. The proposed inverse model uses particle swarm optimization (PSO) algorithm, which is based on the social behavior of animals searching for food sources. The finite-difference numerical solution of the forward analytical model is integrated with the PSO algorithm to solve the inverse problem. The method is tested on synthetic data of base excess pore pressure dissipation curves generated using the finite difference method. The effectiveness of the method is verified using base excess pore pressure dissipation curve obtained from a settling column experiment and further ensured through comparison with available predicted hydraulic conductivity parameters.

Keywords: base excess pore pressure, hydraulic conductivity, large strain consolidation, mine tailings

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929 Determination of Heavy Metal Levels in Carissa spinarum and Toddalia asiatica Used as Herbal Medicines in Kisii and Nyamira Counties Region, Kenya

Authors: Moses A. Guto Maobe, Leonard Gitu, Erastus Gatebe

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The plants Carissa spinarum and Toddalia asiatica have historically been used as herbal medicines in Kisii and Nyamira Counties region, Kenya. But, there is limited study about heavy metal contents in their different plant parts. Such information is necessary for proper use of the two plant species as herbal medicines. So, precise determination of heavy metal contents in different part of these herbs is required for quality, efficacy and safety use in the treatment of ailments. The main aim of this study was to standardize the two herbs of interest. The objective of this study was to evaluate the levels of heavy metal contents in the root of Carissa spinarum and Toddalia asiatica. A wet digestion method with concentrated nitric-hydrochloric acid was used for the dissolution of each herb part prior to elemental analysis. Standard solutions of various concentrations of each pure metal of analytical grade arsenic (As), cadmium (Cd) and mercury (Hg) were prepared and used. The analysis of As, Cd and Hg in each of two herbs was conducted by atomic absorption spectroscopy (AAS) Shimadzu model No. 6200. Data obtained from root of Carissa spinarum indicated concentration (mgkg⁻¹) of Arsenic (As), Cadmium (Cd) and Mercury (Hg) were 0.87 x 10⁻³, 7.02 x 10⁻⁶ and 0.66 x 10⁻³ respectively. Results obtained from root of Toddalia asiatica showed concentration (mgkg⁻¹) of Arsenic (As), Cadmium (Cd) and Mercury (Hg) were 1.33 x 10⁻³, 7.32 x 10⁻⁶ and 1.13 x 10⁻³, respectively. The permissible limits set by WHO for As, Cd and Hg in herbs are (mgkg⁻¹) < 1 - 5, < 0.3 – 1 and < 0.1- 0.5 respectively. The concentrations of As, Cd, and Hg determined were relatively higher in the root of Toddalia asiatica than the root of Carissa spinarum. It was concluded that levels of heavy metal contents of As, Cd, and Hg in the root of Carissa spinarum and Toddalia asiatica were within permissible limits set by WHO/FAO.

Keywords: heavy metals, Carissa spinarum, Toddalia asiatica, wet digestion, pollutants, AAS

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928 Modeling of Drug Distribution in the Human Vitreous

Authors: Judith Stein, Elfriede Friedmann

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The injection of a drug into the vitreous body for the treatment of retinal diseases like wet aged-related macular degeneration (AMD) is the most common medical intervention worldwide. We develop mathematical models for drug transport in the vitreous body of a human eye to analyse the impact of different rheological models of the vitreous on drug distribution. In addition to the convection diffusion equation characterizing the drug spreading, we use porous media modeling for the healthy vitreous with a dense collagen network and include the steady permeating flow of the aqueous humor described by Darcy's law driven by a pressure drop. Additionally, the vitreous body in a healthy human eye behaves like a viscoelastic gel through the collagen fibers suspended in the network of hyaluronic acid and acts as a drug depot for the treatment of retinal diseases. In a completely liquefied vitreous, we couple the drug diffusion with the classical Navier-Stokes flow equations. We prove the global existence and uniqueness of the weak solution of the developed initial-boundary value problem describing the drug distribution in the healthy vitreous considering the permeating aqueous humor flow in the realistic three-dimensional setting. In particular, for the drug diffusion equation, results from the literature are extended from homogeneous Dirichlet boundary conditions to our mixed boundary conditions that describe the eye with the Galerkin's method using Cauchy-Schwarz inequality and trace theorem. Because there is only a small effective drug concentration range and higher concentrations may be toxic, the ability to model the drug transport could improve the therapy by considering patient individual differences and give a better understanding of the physiological and pathological processes in the vitreous.

Keywords: coupled PDE systems, drug diffusion, mixed boundary conditions, vitreous body

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927 Development of Cost-Effective Protocol for Preparation of Dehydrated Paneer (Indian Cottage Cheese) Using Freeze Drying

Authors: Sadhana Sharma, P. K. Nema, Siddhartha Singha

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Paneer or Indian cottage cheese is an acid and heat coagulated milk product, highly perishable because of high moisture (58-60 %). Typically paneer is marble to light creamy white in appearance. A good paneer should have cohesive body with slight sponginess or springiness. The texture must be smooth and velvety with close-knit compactness. It should have pleasing mild acidic, slightly sweet and nutty flavour. Consumers today demand simple to prepare, convenient, healthy and natural foods. Dehydrated paneer finds numerous ways to be used. It can be used in curry preparation similar to paneer-in-curry, a delicacy in Indian cuisine. It may be added to granola/ trail mix yielding a high energy snack. If grounded to a powder, it may be used as a cheesy spice mix or used as popcorn seasoning. Dried paneer powder may be added to pizza dough or to a white sauce to turn it into a paneer sauce. Drying of such food hydrogels by conventional methods is associated with several undesirable characteristics including case hardening, longer drying time, poor rehydration ability and fat loss during drying. The present study focuses on developing cost-effective protocol for freeze-drying of paneer. The dehydrated product would be shelf-stable and can be rehydrated to its original state having flavor and texture comparable to the fresh form. Moreover, the final product after rehydration would be more fresh and softer than its frozen counterparts. The developed product would be shelf-stable at room temperature without any addition of preservatives.

Keywords: color, freeze-drying, paneer, texture

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926 Melatonin Rescue Fungicide Induced Behavioral and Reproductive Abnormalities through Changes of Dopaminergic Activity in the Brain of Catfish, Mystus cavasisu

Authors: Muhammad Badruzzaman, Alif Hasan, Md. Shahjahan

Abstract:

Propiconazole is a triazole fungicide extensively used in agriculture which can harm to non-target organisms in aquatic environment through runoff. Chronic exposure to environmental pesticides turn to behavioral impairment in vertebrates including teleosts. However, the potential effect of this fungicide on neurobehavioral impairment and release from it in vertebrates has not been fully explored. In this work, we examined the role of melatonin to rescue fungicide induced neurobehavioral and reproductive alternation and its connection with changes in dopaminergic activity in the brain of Mystus cavasius. After fish were exposed to water containing propiconazole at 0, 0.1, 5, and 250 µg/L for 3 days, significant increases of DA, 3,4-dihydroxyphenylacetic acid (DOPAC; a DA metabolite), and their ratio (DOPAC/DA) were observed in whole brain at 250 µg/L concentration. When fish were treated with propiconazole at 250 µg/L for 3 days, there was a significant elevation of DA, DOPAC and DOPAC/DA in diencephalon and pituitary, and only DA in the telencephalon, compared with control fish. Besides, it induced a reduction in extracellular serotonin and had an anxiolytic-like effect, supported by a decrease in cortisol production. Increased locomotor activity, anxiety and aggressiveness, decreased gonadosomatic index with few vitellogenic oocytes in ovaries after propiconazole treatment. When fish were treated with melatonin, D1 (SCH-23390) or D2 (Haloperidol) dopamine receptor antagonists and combined of melatonin and D1/D2 receptor antagonist and was observed melatonin + D2 receptor antagonist rescued fungicide induced all behavioral changes in fish. These results indicate that propiconazole increases locomotor activity, anxiety and aggressiveness and decreases reproductive activity, which was rescued by combined treatment of melatonin and dopamine receptor antagonist.

Keywords: behavior, catfish, dopamine, fungicide, melatonin

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925 Impact of Length of Straw by the Use of a Straw Mill on the Selective Feeding of Young Cattle and Their Effects for the Cattle

Authors: Heiko Scholz

Abstract:

When feeding high qualitysilagetoheifersfromthe age of two, there is a riskofenergyoversupply. Depending on the feeding valueorscarceavailability ofsilageorcorn silage diets withhighproportionsof straw is often incorporated. Foran energetically standardized young cattle supply of strawproportion can be more than 20% of dry matter. It was investigated whether the grinding of straw with the strawmillselective feeding significantly limits. The investigation has been carried out with young cattle in the second year. 78 animals were kept and fed under similar conditions in two groups. The experimental group (EG) consisted of cattle 12 to 15 months, and in the control group (CG), the cattle were 15 to 20 months old. The experimental feeding took place in five days of feed distribution, and residual feed were weighed. The ration of EG contained ground with the straw mill straw, and CG was further fed rotor-cut pressed straw. To determine the selective seizure samples of feed distributionandtheremainingfood with the particle separator boxandthecrude protein-and energy-content have been determined. The grinding of the straw increased the daily feed intake.IntheEGan increase infeed intakewas observedby grinding of the straw. Feed intakedirectlyon the day for changing the dietoflongonground straw increased by more than 2.0 kgofDMper animal. In the following days, the feed intakewasincreasedby 0.9kg DMper animal and day on average (7.4 vs. 8.3 kg DM per day). The results of the screen distribution of residual feed point to a differentiated feeding behavior between the groups. In the EG, the particle length of the residual feed to a large extent with the template matches. The acid-base-balance (NSBA)valuesofEGarewithin normal limits. Ifstrawsharesof25% and more are federations to young cattle (heifers), the theparticlelengthof straw has significant impact ontheselectivefeeding behavior. Aparticlelength of 1.5cmcompared to7.5 cmlongpreventedstrawcertainly discarding of the straw on the feeding barn. The feed intake increases whenshortstrawis mixed into theTMR.

Keywords: straw mill, heifer, feed selection, dry matter intake

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924 Effect of Oxytocin on Cytosolic Calcium Concentration of Alpha and Beta Cells in Pancreas

Authors: Rauza Sukma Rita, Katsuya Dezaki, Yuko Maejima, Toshihiko Yada

Abstract:

Oxytocin is a nine-amino acid peptide synthesized in the paraventricular nucleus (PVN) and supraoptic nucleus (SON) of the hypothalamus. Oxytocin promotes contraction of the uterus during birth and milk ejection during breast feeding. Although oxytocin receptors are found predominantly in the breasts and uterus of females, many tissues and organs express oxytocin receptors, including the pituitary, heart, kidney, thymus, vascular endothelium, adipocytes, osteoblasts, adrenal gland, pancreatic islets, and many cell lines. On the other hand, in pancreatic islets, oxytocin receptors are expressed in both α-cells and β-cells with stronger expression in α- cells. However, to our knowledge there are no reports yet about the effect of oxytocin on cytosolic calcium reaction on α and β-cell. This study aims to investigate the effect of oxytocin on α-cells and β-cells and its oscillation pattern. Islet of Langerhans from wild type mice were isolated by collagenase digestion. Isolated and dissociated single cells either α-cells or β-cells on coverslips were mounted in an open chamber and superfused in HKRB. Cytosolic concentration ([Ca2+]i) in single cells were measured by fura-2 microfluorimetry. After measurement of [Ca2+]i, α-cells were identified by subsequent immunocytochemical staining using an anti-glucagon antiserum. In β-cells, the [Ca2+]i increase in response to oxytocin was observed only under 8.3 mM glucose condition, whereas in α-cells, [Ca2+]i an increase induced by oxytocin was observed in both 2.8 mM and 8.3 mM glucose. The oscillation incidence was induced more frequently in β-cells compared to α-cells. In conclusion, the present study demonstrated that oxytocin directly interacts with both α-cells and β-cells and induces increase of [Ca2+]i and its specific patterns.

Keywords: α-cells, β-cells, cytosolic calcium concentration, oscillation, oxytocin

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923 Effect of Probiotic Feeding on Weight Gain, Blood Biochemical and Hematological Indices of Crossbred Dairy Goat Kids

Authors: Claire B. Salvedia, Enrico P. Supangco, Francisco B. Eligado, Renato Sa Vega, Antonio A. Rayos

Abstract:

The study was conducted to evaluate the effect of probiotic feeding on weight gain, blood biochemical and hematological indices of crossbred dairy goat kids. Sixteen (16) crossbred Anglo-Nubian x Saanen dairy goat kids, 3 to 4 months old, ranging from 19 to 23kg were randomly assigned into four treatments fed with 5x109 cfu/ml probiotic supplements; Treatment 1 – control; Treatment 2 – lactic acid bacteria (L. plantarum BS and P. acidilactici 3G3); treatment 3 – S. cerevisiae 2030; Treatment 4 – multi-strain probiotics (L. plantarum BS, P. acidilactici 3G3, and S.cerevisiae 2030). Feed ration provided daily for each of the experimental animals were composed of 1kg mixed concentrate feed ((Leucaena leucocephala dried leaves and pollard), and 4 kg fresh Pennisetum purpureum and Gliciridia sepium leaves (50:50). The experimental feeding trial lasted for 9 weeks. Result revealed that treatments fed with probiotics had significantly (P≤0.05) higher weight gain compared to the control. Significant effect on plasma urea nitrogen (PUN) and triglyceride were noted during 30th and 60th day of probiotic feeding. White blood cell counts were significantly affected by probiotic feeding during the 60th day. Concentrations of glucose and cholesterol remained unchanged throughout the experimental period. The findings suggests, under the condition of the experiment, that live probiotic feeding could have a significant role in improving weight gain and metabolism of crossbred dairy goat kids.

Keywords: probiotics, weight gain, blood biochemical indices, crossbred dairy goat kids

Procedia PDF Downloads 492
922 Characterization of Fungal Endophytes in Leaves, Stems and Roots of African Yam Bean (Sphenostylis sternocarpa Hochst ex. A. Rich Harms)

Authors: Iyabode A. Kehinde, Joshua O. Oyekanmi, Jumoke T. Abimbola, Olajumoke E. Ayanda

Abstract:

African yam bean (AYB), (Sphenostylis stenocarpa) is a leguminous crop that provides nutritionally rich seeds, tubers and leaves for human consumption. AYB potentials as an important food security crop is yet to be realized and thus classified as underutilized crop. Underutilization of the crop has been partly associated with scarce information on the incidence and characterization of fungal endophytes infecting vascular parts of AYB. Accurate and robust detection of these endophytic fungi is essential for diagnosis, modeling, surveillance and protection of germplasm (seed) health. This work aimed at isolating and identifying fungal endophytes associated with leaves, stems and roots of AYB in Ogun State, Nigeria. This study investigated both cultural and molecular properties of endophytic fungi in AYB for its characterization and diversity. Fungal endophytes were isolated and culturally identified. DNA extraction, PCR amplification using ITS primers and analyses of nucleotide sequences of ribosomal DNA fragments were conducted on selected isolates. BLAST analysis was conducted on consensus nucleotide sequences of 28 out of 30 isolates and results showed similar homology with genera of Rhizopus, Cunninghamella, Fusarium, Aspergillus, Penicillium, Alternaria, Diaporthe, Nigrospora, Purpureocillium, Corynespora, Magnaporthe, Macrophomina, Curvularia, Acrocalymma, Talaromyces and Simplicillium. Slight similarity was found with endophytes associated with soybean. Phylogenetic analysis by maximum likelihood method showed high diversity among the general. These organisms have high economic importance in crop improvement. For an instance, Purpureocillium lilacinum showed high potential in control of root rot caused by nematodes in tomatoes. Though some can be pathogens, but many of the fungal endophytes have beneficial attributes to plant in host health, uptake of nutrients, disease suppression, and host immunity.

Keywords: molecular characterization, African Yam Bean, fungal endophyte, plant parts

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921 Reducing Antimicrobial Resistance Using Biodegradable Polymer Composites of Mof-5 for Efficient and Sustained Delivery of Cephalexin and Metronidazole

Authors: Anoff Anim, Lila Mahmound, Maria Katsikogianni, Sanjit Nayak

Abstract:

Sustained and controlled delivery of antimicrobial drugs have been largely studied recently using metal organic frameworks (MOFs)and different polymers. However, much attention has not been given to combining both MOFs and biodegradable polymers which would be a good strategy in providing a sustained gradual release of the drugs. Herein, we report a comparative study of the sustained and controlled release of widely used antibacterial drugs, cephalexin and metronidazole, from zinc-based MOF-5 incorporated in biodegradable polycaprolactone (PCL) and poly-lactic glycolic acid (PLGA) membranes. Cephalexin and metronidazole were separately incorporated in MOF-5 post-synthetically, followed by their integration into biodegradable PLGA and PCL membranes. The pristine MOF-5 and the loaded MOFs were thoroughly characterized by FT-IR, SEM, TGA and PXRD. Drug release studies were carried out to assess the release rate of the drugs in PBS and distilled water for up to 48 hours using UV-Vis Spectroscopy. Four bacterial strains from both the Gram-positive and Gram-negative types, Staphylococus aureus, Staphylococuss epidermidis, Escherichia coli, Acinetobacter baumanii, were tested against the pristine MOF, pure drugs, loaded MOFs and the drug-loaded MOF-polymer composites. Metronidazole-loaded MOF-5 composite of PLGA (PLGA-Met@MOF-5) was found to show highest efficiency to inhibit the growth of S. epidermidis compared to the other bacteria strains while maintaining a sustained minimum inhibitory concentration (MIC). This study demonstrates that the combination of biodegradable MOF-polymer composites can provide an efficient platform for sustained and controlled release of antimicrobial drugs, and can be a potential strategy to integrate them in biomedical devices.

Keywords: antimicrobial resistance, biodegradable polymers, cephalexin, drug release metronidazole, MOF-5, PCL, PLGA

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920 Physical and Microbiological Evaluation of Chitosan Films: Effect of Essential Oils and Storage

Authors: N. Valderrama, W. Albarracín, N. Algecira

Abstract:

It was studied the effect of the inclusion of thyme and rosemary essential oils into chitosan films, as well as the microbiological and physical properties when storing chitosan film with and without the mentioned inclusion. The film forming solution was prepared by dissolving chitosan (2%, w/v), polysorbate 80 (4% w/w CH) and glycerol (16% w/w CH) in aqueous lactic acid solutions (control). The thyme (TEO) and rosemary (REO) essential oils (EOs) were included 1:1 w/w (EOs:CH) on their combination 50/50 (TEO:REO). The films were stored at temperatures of 5, 20, 33°C and a relative humidity of 75% during four weeks. The films with essential oil inclusion did not show an antimicrobial activity against strains. This behavior could be explained because the chitosan only inhibits the growth of microorganisms in direct contact with the active sites. However, the inhibition capacity of TEO was higher than the REO and a synergic effect between TEO:REO was found for S. enteritidis strains in the chitosan solution. Some physical properties were modified by the inclusion of essential oils. The addition of essential oils does not affect the mechanical properties (tensile strength, elongation at break, puncture deformation), the water solubility, the swelling index nor the DSC behavior. However, the essential oil inclusion can significantly decrease the thickness, the moisture content, and the L* value of films whereas the b* value increased due to molecular interactions between the polymeric matrix, the loosing of the structure, and the chemical modifications. On the other hand, the temperature and time of storage changed some physical properties on the chitosan films. This could have occurred because of chemical changes, such as swelling in the presence of high humidity air and the reacetylation of amino groups. In the majority of cases, properties such as moisture content, tensile strength, elongation at break, puncture deformation, a*, b*, chrome, ΔE increased whereas water resistance, swelling index, L*, and hue angle decreased.

Keywords: chitosan, food additives, modified films, polymers

Procedia PDF Downloads 366
919 Effect of a Polyherbal Gut Therapy Protocol in Changes of Gut and Behavioral Symptoms of Antibiotic Induced Dysbiosis of Autistic Babies

Authors: Dinesh K. S., D. R. C. V. Jayadevan

Abstract:

Autism is the most prevalent of a subset of the disorders organized under the umbrella of pervasive developmental disorders. After the publication of Andrew Wakefield's paper in lancet, many critiques deny this connection even without looking in to the matter. The British Medical Journal even put an editorial regarding this issue. BMJ 2010; 340:c1807. But ayurveda has ample of evidences to believe this connectivity. Dysbiosis, yeast growth of the gut, nutritional deficiencies, enzyme deficiencies, essential fatty acid deficiencies, Gastro esophageal reflux disease, indigestion, inflammatory bowel, chronic constipation & its cascade are few of them to note. The purpose of this paper is to present the observed changes in the behavioural symptoms of autistic babies after a gut management protocol which is a usual programme of our autism treatment plan especially after dysbiotic changes after antibiotic administration. Is there any correlation between changes (if significant) in gut symptoms and behavioral problems of autistic babies especially after a dysbiosis induced by antibiotics. Retrospective analysis of the case sheets of autistic patients admitted in Vaidyaratnam P.S.Varier Ayurveda College hospital, kottakkal,kerala, india from September 2010 are taken for the data processing. Autistic patients are used to come to this hospital as a part of their usual course of treatment. We investigated 40 cases diagnosed as autistic by clinical psychologists from different institutions who had dysbiosis induced by antibiotics. Significant change in gut symptoms before and after treatment p<0.05 in most of its components Significant change in behavioral symptoms before and after treatments p<0.05 in most of the components Correlation between gut symptoms change and behavioral symptoms changes after treatment is + 0.86. Conclusion : Selected Polyherbal Ayurveda treatment has significant role to play to make changes abnormal behaviors in autistic babies and has a positive correlation with changes in gut symptoms induced by dysbiosis of antibiotic intake.

Keywords: ayurveda, autism, dysbiosis, antibiotic

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918 Isolation, Characterization and Screening of Antimicrobial Producing Actinomycetes from Sediments of Persian Gulf

Authors: H. Alijani, M. Jabari, S. Matroodi, H. Zolqarnein, A. Sharafi, I. Zamani

Abstract:

Actinomycetes, Gram-positive bacteria, are interesting as a main producer of secondary metabolites and are important industrially and pharmaceutically. The marine environment is a potential source for new actinomycetes, which can provide novel bioactive compounds and industrially important enzymes. The aims of this study were to isolate and identify novel actinomycetes from Persian Gulf sediments and screen these isolates for the production of secondary metabolites, especially antibiotics, Using phylogenetic (16S rRNA gene sequence), morphological and biochemical analyses. 15 different actinomycete strains from Persian Gulf sediments at a depth of 5-10 m were identified. DNA extraction was done using Cinnapure DNA Kit. PCR amplification of 16S rDNA gene was performed using F27 and R1492 primers. Phylogenetic tree analysis was performed using the MEGA 6 software. Most of the isolated strains belong to the genus namely Streptomyces (14), followed by Nocardiopsis (1). Antibacterial assay of the isolates supernatant was performed using a standard disc diffusion assay with replication (n=3). The results of disk diffusion assay showed that most active strain against Proteus volgaris and Bacillus cereus was AMJ1 (16.46±0.2mm and 13.78±0.2mm, respectively), against Salmonella sp. AMJ7 was the most effective strain (10.13±0.2mm), and AMJ1 and AHA5 showed more inhibitory activity against Escherichia coli (8.04±0.02 mm and 8.2±0.03 ). The AMJ6 strain showed best antibacterial activity against Klebsiella sp. (8.03±0.02mm). Antifungal activity of AMJ2 showed that it was most active strain against complex (16.05±0.02mm) and against Aspergillus flavus strain AMJ1 was most active strain (16.4±0.2mm) and highest antifungal activity against Trichophyton mentagrophytes, Microsporum gyp serum and Candida albicans, were shown by AHA1 (21.03±0.02mm), AHA3 and AHA7 (18±0.03mm), AMJ6 (21.03±0.2mm) respectively. Our results revealed that the marine actinomycetes of Persian Gulf sediments were potent source of novel antibiotics and bioactive compounds and indicated that the antimicrobial metabolites were extracellular. Most of the secondary metabolites and antibiotics are extracellular in nature and extracellular products of actinomycetes show potent antimicrobial activities.

Keywords: antibacterial activity, antifungal activity, marine actinomycetes, Persian Gulf

Procedia PDF Downloads 297
917 Electrochemical Growth and Properties of Cu2O Nanostructures

Authors: A. Azizi, S. Laidoudi, G. Schmerber, A. Dinia

Abstract:

Cuprous oxide (Cu2O) is a well-known oxide semiconductor with a band gap of 2.1 eV and a natural p-type conductivity, which is an attractive material for device applications because of its abundant availability, non toxicity, and low production cost. It has a higher absorption coefficient in the visible region and the minority carrier diffusion length is also suitable for use as a solar cell absorber layer and it has been explored in junction with n type ZnO for photovoltaic applications. Cu2O nanostructures have been made by a variety of techniques; the electrodeposition method has emerged as one of the most promising processing routes as it is particularly provides advantages such as a low-cost, low temperature and a high level of purity in the products. In this work, Cu2O nanostructures prepared by electrodeposition from aqueous cupric sulfate solution with citric acid at 65°C onto a fluorine doped tin oxide (FTO) coated glass substrates were investigated. The effects of deposition potential on the electrochemical, surface morphology, structural and optical properties of Cu2O thin films were investigated. During cyclic voltammetry experiences, the potential interval where the electrodeposition of Cu2O is carried out was established. The Mott–Schottky (M-S) plot demonstrates that all the films are p-type semiconductors, the flat-band potential and the acceptor density for the Cu2O thin films are determined. AFM images reveal that the applied potential has a very significant influence on the surface morphology and size of the crystallites of thin Cu2O. The XRD measurements indicated that all the obtained films display a Cu2O cubic structure with a strong preferential orientation of the (111) direction. The optical transmission spectra in the UV-Visible domains revealed the highest transmission (75 %), and their calculated gap values increased from 1.93 to 2.24 eV, with increasing potentials.

Keywords: Cu2O, electrodeposition, Mott–Schottky plot, nanostructure, optical properties, XRD

Procedia PDF Downloads 355
916 Structural and Functional Comparison of Untagged and Tagged EmrE Protein

Authors: S. Junaid S. Qazi, Denice C. Bay, Raymond Chew, Raymond J. Turner

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EmrE, a member of the small multidrug resistance protein family in bacteria is considered to be the archetypical member of its family. It confers host resistance to a wide variety of quaternary cation compounds (QCCs) driven by proton motive force. Generally, purification yield is a challenge in all membrane proteins because of the difficulties in their expression, isolation and solubilization. EmrE is extremely hydrophobic which make the purification yield challenging. We have purified EmrE protein using two different approaches: organic solvent membrane extraction and hexahistidine (his6) tagged Ni-affinity chromatographic methods. We have characterized changes present between ligand affinity of untagged and his6-tagged EmrE proteins in similar membrane mimetic environments using biophysical experimental techniques. Purified proteins were solubilized in a buffer containing n-dodecyl-β-D-maltopyranoside (DDM) and the conformations in the proteins were explored in the presence of four QCCs, methyl viologen (MV), ethidium bromide (EB), cetylpyridinium chloride (CTP) and tetraphenyl phosphonium (TPP). SDS-Tricine PAGE and dynamic light scattering (DLS) analysis revealed that the addition of QCCs did not induce higher multimeric forms of either proteins at all QCC:EmrE molar ratios examined under the solubilization conditions applied. QCC binding curves obtained from the Trp fluorescence quenching spectra, gave the values of dissociation constant (Kd) and maximum specific one-site binding (Bmax). Lower Bmax values to QCCs for his6-tagged EmrE shows that the binding sites remained unoccupied. This lower saturation suggests that the his6-tagged versions provide a conformation that prevents saturated binding. Our data demonstrate that tagging an integral membrane protein can significantly influence the protein.

Keywords: small multidrug resistance (SMR) protein, EmrE, integral membrane protein folding, quaternary ammonium compounds (QAC), quaternary cation compounds (QCC), nickel affinity chromatography, hexahistidine (His6) tag

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915 Chemical Composition and Nutritional Value of Leaves and Pods of Leucaena Leucocephala, Prosopis Laevigata and Acacia Farnesiana in a Xerophyllous Shrubland

Authors: Miguel Mellado, Cecilia Zapata

Abstract:

Goats can be exploited in harsh environments due to their capacity to adjust to limited quantity and quality forage sources. In these environments, leguminous trees can be used as supplementary feeds as foliage and fruits of these trees can contribute to maintain or improve production efficiency in ruminants. The objective of this study was to determine the nutritional value of three leguminous trees heavily selected by goats in a xerophyllous shrubland. Chemical composition and in vitro dry matter disappearance (IVDMD) of leaves and pods from leucaena (Leucaena leucocephala), mesquite (Prosopis laevigata) and huisache (Acacia farnesiana) is presented. Crude protein (CP) ranged from 17.3% for leaves of huisache to 21.9% for leucaena. The neutral detergent fiber (NDF) content ranged from 39.0 to 40.3 with no difference among fodder threes. Across tree species, mean IVDMD was 61.6% for pods and 52.2% for leaves. IVDMD for leaves was highest (P < 0.01) for leucaena (54.9%) and lowest for huisache (47.3%). Condensed tannins in an acetonic extract were highest for leaves of huisache (45.3 mg CE/g DM) and lowest for mesquite (25.9 mg CE/g DM). Pods and leaves of huisache presented the highest number of secondary metabolites, mainly related to hydrobenzoic acid and flavonols; leucaena and mesquite presented mainly flavonols and anthocyanins. It was concluded that leaves and pods of leucaena, mesquite and huisache constitute valuable forages for ruminant livestock due to its low fiber, high CP levels, moderate in vitro fermentation characteristics and high mineral content. Keywords: Fodder tree; ruminants; secondary metabolites; minerals; tannins

Keywords: fodder tree, ruminants, secondary metabolites, minerals, tannins

Procedia PDF Downloads 145
914 Nanopharmaceutical: A Comprehensive Appearance of Drug Delivery System

Authors: Mahsa Fathollahzadeh

Abstract:

The various nanoparticles employed in drug delivery applications include micelles, liposomes, solid lipid nanoparticles, polymeric nanoparticles, functionalized nanoparticles, nanocrystals, cyclodextrins, dendrimers, and nanotubes. Micelles, composed of amphiphilic block copolymers, can encapsulate hydrophobic molecules, allowing for targeted delivery. Liposomes, vesicular structures made up of phospholipids, can encapsulate both hydrophobic and hydrophilic molecules, providing a flexible platform for delivering therapeutic agents. Solid lipid nanoparticles (SLNs) and nanostructured lipid carriers (NLCs) are designed to improve the stability and bioavailability of lipophilic drugs. Polymeric nanoparticles, such as poly(lactic-co-glycolic acid) (PLGA), are biodegradable and can be engineered to release drugs in a controlled manner. Functionalized nanoparticles, coated with targeting ligands or antibodies, can specifically target diseased cells or tissues. Nanocrystals, engineered to have specific surface properties, can enhance the solubility and bioavailability of poorly soluble drugs. Cyclodextrins, doughnut-shaped molecules with hydrophobic cavities, can be complex with hydrophobic molecules, allowing for improved solubility and bioavailability. Dendrimers, branched polymers with a central core, can be designed to deliver multiple therapeutic agents simultaneously. Nanotubes and metallic nanoparticles, such as gold nanoparticles, offer real-time tracking capabilities and can be used to detect biomolecular interactions. The use of these nanoparticles has revolutionized the field of drug delivery, enabling targeted and controlled release of therapeutic agents, reduced toxicity, and improved patient outcomes.

Keywords: nanotechnology, nanopharmaceuticals, drug-delivery, proteins, ligands, nanoparticles, chemistry

Procedia PDF Downloads 51
913 Magnetophotonics 3D MEMS/NEMS System for Quantitative Mitochondrial DNA Defect Profiling

Authors: Dar-Bin Shieh, Gwo-Bin Lee, Chen-Ming Chang, Chen Sheng Yeh, Chih-Chia Huang, Tsung-Ju Li

Abstract:

Mitochondrial defects have a significant impact in many human diseases and aging associated phenotypes. The pathogenic mitochondrial DNA (mtDNA) mutations are diverse and usually present as heteroplasmic. mtDNA 4977bps deletion is one of the common mtDNA defects, and the ratio of mutated versus normal copy is significantly associated with clinical symptoms thus their quantitative detection has become an important unmet needs for advanced disease diagnosis and therapeutic guidelines. This study revealed a Micro-electro-mechanical-system (MEMS) enabled automatic microfluidic chip that only required minimal sample. The system integrated multiple laboratory operation steps into a Lab-on-a-Chip for high-sensitive and prompt measurement. The entire process including magnetic nanoparticle based mtDNA extraction in chip, mutation selective photonic DNA cleavage, and nanoparticle accelerated photonic quantitative polymerase chain reaction (qPCR). All subsystems were packed inside a miniature three-dimensional micro structured system and operated in an automatic manner. Integration of magnetic beads with microfluidic transportation could promptly extract and enrich the specific mtDNA. The near infrared responsive magnetic nanoparticles enabled micro-PCR to be operated by pulse-width-modulation controlled laser pulsing to amplify the desired mtDNA while quantified by fluorescence intensity captured by a complementary metal oxide system array detector. The proportions of pathogenic mtDNA in total DNA were thus obtained. Micro capillary electrophoresis module was used to analyze the amplicone products. In conclusion, this study demonstrated a new magnetophotonic based qPCR MEMS system that successfully detects and quantify specific disease related DNA mutations thus provides a promising future for rapid diagnosis of mitochondria diseases.

Keywords: mitochondrial DNA, micro-electro-mechanical-system, magnetophotonics, PCR

Procedia PDF Downloads 218