Search results for: isolated bridges
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1904

Search results for: isolated bridges

434 Ring FingerPortein 2 (RNF2) Targeting by miRNAs in Breast Cancer Cell Lines

Authors: Ceyda Okudu, Secil Eroglu, Khandakar A. S. M. Saadat, Sibel O. Balci

Abstract:

Ring Finger Protein 2 (RNF2) is a member of polycomb repressive complex 1 (PRC1), which is one of the epigenetic regulators in the genome. When RNF2 combines with other PRC1 members, it mediates the mono-ubiquitination of Histon2A (H2A). In breast cancer, RNF2 is commonly overexpressed, and also it promotes metastasis and invasion in other aggressive tumors like melanoma, prostate, and hepatocarcinoma. The role of RNF2 in the metastasis and invasion of breast cancer has not yet been elucidated. Our aim is to observe the role of RNF2 in metastasis and invasion in this study by miRNA mediated RNF2 gene silencing in breast cancer cell lines. We selected miRNAs, targeting to RNF2 by searching online databases. miR-17-5p, miR20a-5p, and miR-106b-5p were transfected to breast cancer cell lines (MCF-7, MDA-MB-231, SK-BR-3, and ZR-75-1), and also we used normal breast epithelial cell line (hTERT-HME1) to compare RNF2 gene expression level. After 48-72 hours post-transfection, mRNAs were isolated from the cells, and gene expressions were measured by RT-qPCR after from cDNA syntheses. We observed that RNF2 was highly expressed in SK-BR-3 and MDA-MB-231 cell lines opposite to MCF-7 and ZR-75-1 cell lines. RNF2 was downregulated 5, 5 and 7 fold by miR17-5p, miR20a-5p and miR106b-5p respectively in MCF-7. However, in SK-BR-3 and ZR-75-1 cell lines, miRNAs did not affect significantly RNF2 gene expression level. miR20a-5p decreased RNF2 3 fold and miR17-5p and miR106b-5p did not affect MDA-MB-231. After gene expression analysis, we performed metastasis and invasion assay in MCF-7 cells. For metastasis, we used both wound healing assay and Transwell Cell Migration Assay, and we used Transwell Cell Invasion Assay for invasion. The data of this assay showed that miR17-5p and miR20a-5p decreased both invasion and metastasis level, but miR106b-5p has no effect. We would like to conclude that RNF2 can be targeted by miR17-5p, miR20a-5p and miR106b-5p in MCF-7 cells and also RNF2, which is one of the upregulated genes in aggressive tumor, can be decreased by using these miRNAs. In future, we would like to confirm these results at the protein level and also whether these miRNAs are direct target of RNF2 or not.

Keywords: breast cancer, epigenetic, microRNAs, RNF2

Procedia PDF Downloads 156
433 The Effect of Increased Tip Area of Suction Caissons on the Penetration Resistance Coefficients

Authors: Ghaem Zamani, Farveh Aghaye Nezhad, Amin Barari

Abstract:

The installation process of caissons has usually been a challenging step in the design phase, especially in the case of suction-assisted installation. The engineering practice for estimating the caisson penetration resistance is primarily controlled by the resistance governed by inner and outer skirt friction and the tip resistance. Different methods have been proposed in the literature to evaluate the above components, while the CPT-based methodology has attained notable popularity among others. In this method, two empirical coefficients are suggested, k𝒻 and kp, which relate the frictional resistance and tip resistance to the cone penetration resistance (q𝒸), respectively. A series of jacking installation and uninstallation experiments for different soil densities were carried out in the offshore geotechnical laboratory of Aalborg University, Denmark. The main goal of these tests was to find appropriate values for empirical coefficients of the CPT-based method for the buckets with large embedment ratio (i.e., d/D=1, where d is the skirt length and D is the diameter) and increased tip area penetrated into dense sand deposits. The friction resistance effects were isolated during the pullout experiments; hence, the k𝒻 was back-measured from the tests in the absence of tip resistance. The actuator force during jacking installation equals the sum of frictional resistance and tip resistance. Therefore, the tip resistance of the bucket is calculated by subtracting the back-measured frictional resistance from penetration resistance; hence the relevant coefficient kp would be achieved. The cone penetration test was operated at different points before and after each installation attempt to measure the cone penetration resistance (q𝒸), and the average value of q𝒸 is used for calculations. The experimental results of the jacking installation tests indicated that a larger friction area considerably increased the penetration resistance; however, this effect was completely diminished when foundation suction-assisted penetration was used. Finally, the values measured for the empirical coefficient of the CPT-based method are compared with the highest expected and most probable values suggested by DNV(1992) for uniform thickness buckets.

Keywords: suction caisson, offshore geotechnics, cone penetration test, wind turbine foundation

Procedia PDF Downloads 63
432 Pioglitazone Ameliorates Methotrexate-Induced Renal Endothelial Dysfunction via Amending Detrimental Changes in Antioxidant Profile, Systemic Cytokines and Apoptotic Factors

Authors: Sahar M. El-Gowilly, Mai M. Helmy, Hanan M. El-Gowelli

Abstract:

Methotrexate (MTX) is widely used in treatment of cancers and autoimmune diseases. However, nephrotoxicity is one of the most important side effects of MTX. The peroxisome proliferator-activated receptor gamma agonist, pioglitazone (PIO), is known to exert anti-inflammatory and reno-protective effects in various kidney injuries. The purpose of this study was to investigate the potential involvement of endothelial damage in MTX-induced renal injury and to elaborate the possible protective effect of PIO against MTX-induced nephropathy. Compared with saline-treated rats, treatment with MTX (7 mg/kg for 3 day) caused significant elevations in serum levels of urea and creatinine, increased renal nitrate/nitrite level and impaired renovascular responsiveness of isolated perfused kidney to endothelium-dependent vasodilations induced by acetylcholine (0.01-2.43 nmol) and isoprenaline (1µmol). These effects were abolished by concurrent treatment with PIO (2.5 mg/kg, for 5 days starting two days before MTX). Alternatively, MTX treatment did not affect endothelium-independent renovascular relaxation induced by sodium nitroprusside (1-30 μmole). The possibility that alterations in renal antioxidants, circulating cytokine and apoptotic factor (Fas) levels contributed to MTX-PIO interaction was assessed. PIO treatment abrogated renal oxidative stress (decreased reduced glutathione and catalase activity and increased malondialdehyde), elevated serum cytokine (interleukin-6, interleukin-10, tumor necrosis factor-alpha and transforming growth factor-beta1) and Fas induced by MTX. Histologically, MTX caused defused tubular cells swelling and vacuolization associated with endothelial damage in renal arterioles. These effects disappeared upon co-treated with PIO. Collectively, PIO abolished MTX-induced endothelium dysfunction and nephrotoxicity via ameliorating oxidative stress and rectifying cytokines and Fas abnormalities caused by MTX.

Keywords: methotrexate, pioglitazone, endothelium, kidney

Procedia PDF Downloads 296
431 Characterisation of Chitooligomers Prepared with the Aid of Cellulase, Xylanase and Chitosanase

Authors: Anna Zimoch-Korzycka, Dominika Kulig, Andrzej Jarmoluk

Abstract:

The aim of this study was to obtain chitooligosaccharides from chitosan with better functional properties using three different enzyme preparations and compare the products of enzymatic hydrolysis. Commercially available cellulase (CL), xylanase (X) and chitosanase (CS) preparations were used to investigate hydrolytic activity on chitosan (CH) with low molecular weight and DD of 75-85%. It has been reported that CL and X have side activities of other enzymes, such as β-glucanase or β-glucosidase. CS enzyme has a foreign activity of chitinase. Each preparation was used in 1000 U of activity and in the same reaction conditions. The degree of deacetylation and molecular weight of chitosan were specified using titration and viscometric methods, respectively. The hydrolytic activity of enzymes preparations on chitosan was monitored by dynamic viscosity measurement. After 4 h reaction with stirring, solutions were filtered and chitosan oligomers were isolated by methanol solution into two fractions: precipitate (A) and supernatant (B). A Fourier-transform infrared spectroscopy was used to characterize the structural changes of chitosan oligomers fractions and initial chitosan. Furthermore, the solubility of lyophilized hydrolytic mixture (C) and two chitooligomers fractions (A, B) of each enzyme hydrolysis was assayed. The antioxidant activity of chitosan oligomers was evaluated as DPPH free radical scavenging activity. The dynamic viscosity measured after addition of enzymes preparation to the chitosan solution decreased dramatically over time in the sample with X in comparison to solution without the enzyme. For mixtures with CL and CS, lower viscosities were also recorded but not as low as the ones with X. A and B fractions were characterized by the most similar viscosity obtained by the xylanase hydrolysis and were 15 mPas and 9 mPas, respectively. Structural changes of chitosan oligomers A, B, C and their differences related with various enzyme preparations used were confirmed. Water solubility of A fractions was not possible to filter and the result was not recorded. Solubility of supernatants was approximately 95% and was higher than hydrolytic mixture. It was observed that the DPPH radical scavenging effect of A, B, C samples is the highest for X products and was approximately 13, 17, 19% respectively. In summary, a mixture of chitooligomers may be useful for the design of edible protective coatings due to the improved biophysical properties.

Keywords: cellulase, xylanase, chitosanase, chitosan, chitooligosaccharides

Procedia PDF Downloads 309
430 Bioconversion of Kitchen Waste to Bio-Ethanol for Energy Security and Solid Waste Management

Authors: Sanjiv Kumar Soni, Chetna Janveja

Abstract:

The approach of utilizing zero cost kitchen waste residues for growing suitable strains of fungi for the induction of a cocktail of hydrolytic enzymes and ethanol generation has been validated in the present study with the objective of developing an indigenous biorefinery for low cost bioethanol production with the generation of zero waste. Solid state fermentation has been carried out to evaluate the potential of various steam pretreated kitchen waste residues as substrates for the co-production of multiple carbohydrases including cellulases, hemicellulases, pectinase and amylases by a locally isolated strain of Aspergillus niger C-5. Of all the residues, potato peels induced the maximum yields of all the enzyme components corresponding to 64.0±1.92 IU of CMCase, 17.0±0.54 IU of FPase , 42.8±1.28 IU of β-glucosidase, 990.0±28.90 IU of xylanase, 53.2±2.12 IU of mannanase, 126.0±3.72 IU of pectinase, 31500.0±375.78 IU of α-amylase and 488.8±9.82 IU of glucoamylase/g dry substrate respectively. Saccharification of various kitchen refuse residues using inhouse produced crude enzyme cocktail resulted in the release of 610±10.56, 570±8.89, 435±6.54, 475±4.56, 445±4.27, 385±4.49, 370±6.89, 490±10.45 mg of total reducing sugars/g of dried potato peels, orange peels, pineapple peels, mausami peels, onion peels, banana stalks, pea pods and composite mixture respectively revealing carbohydrate conversion efficiencies in the range of 97.0-99.4%. After fermentation of released hexoses by Saccharomyces cerevisae, ethanol yields ranging from 80-262 mL/ kg of dry residues were obtained. The study has successfully evaluated the valorization of kitchen garbage, a highly biodegradable component in Municipal Solid Waste by using it as a substrate for the in-house co-production of multiple carbohydrases and employing the steam treated residues as a feed stock for bioethanol production. Such valorization of kitchen garbage may reduce the level of Municipal Solid Waste going into land-fills thus lowering the emissions of greenhouse gases. Moreover, the solid residue left after the bioconversion may be used as a biofertilizer for improving the fertility of the soils.

Keywords: kitchen waste, bioethanol, solid waste, bioconversion, waste management

Procedia PDF Downloads 379
429 In vitro Antifungal Activity of Methanolic Extracts of Eight Various Cultivar of Persian Punica granatum L. against Candida Species

Authors: Shahindokht Bassiri-Jahromi, Mohammad Reza Pourshafie, Farzad Katiraee, Mannan Hajimahmoodi, Ehsan Mostafavi, Malihe Talebi

Abstract:

Objective: Resistance of Candida species to antifungal agents has potentially serious implications for management of infections. Candida species are now fourth common organisms isolated from hospitalized patients. It is important to increase effective therapy. In the past decade, numerous reports of treatment failures were reported. Prevention and control of these infections will require new antimicrobial agents. Plant-derived antifungal have always been a source of novel therapeutics. The aim of this study was to investigate the antifungal effect of methanolic extract of pomegranate peel and pulp against Candida species. Material and Methods: Eight cultivars of Punica granatum L. were collected from Saveh Agricultural Investigation Center in Iran. Both pomegranate pulp and peel were dried and powdered separately. The dried powders were extracted by using a soxhlet extractor. The antifungal effect of methanolic extract of pomegranate peel and pulp were determined in vitro by minimum inhibitory concentration (MIC) against five standard species of (ATCC 10231), C. parapsilosis (ATCC 22019), C. tropicalis (ATCC 750), C. glabrata (PTCC 5297), and C. kroseii (PTCC 5295). Results: Maximum inhibitions of antifungal effect were attributed to peel extract pomegranate cultivar and Candida species. The most potential antifungal inhibition among 8 different cultivars observed by sour malas, sour white peel, and sour summer extracts respectively, against five Candida strains. The antifungal activity of pulp extracts against Candida species was approximately negative. Conclusion: The use of Punica granatum peel extract has been shown to possess antifungal activities. The phytochemistry and pharmacological actions of Punica granatum peel components suggest a wide range of clinical applications for the treatment and prevention of candidiasis.

Keywords: antifungal activity, Candida species, Punica granatum L., pharmacognosy

Procedia PDF Downloads 458
428 The Anti-Bladder Cancer Effects Exerted by Hyaluronan Nanoparticles Encapsulated Heteronemin Isolated from Hippospongia Sp.

Authors: Kuan Yin Hsiao, Shyh Ming Kuo, Yi Jhen Wu, Chin Wen Chuang, Chuen-Fu Lin, Wei-qing Yang, Han Hsiang Huang

Abstract:

Anti-tumor effects of natural products, like compounds from marine sponges and soft corals, have been investigated for decades. Polymeric nanoparticles prepared from biodegradable and biocompatible molecules, such as Hyaluronan (HA), Chitosan (CHI) and gelatin have been widely studied. Encapsulation of anti-cancer therapies by the biopolymeric nanoparticles in drug delivery system is potentially capable of improving the therapeutic effects and attenuating their toxicity. In the current study, the anti-bladder cancer effects of heteronemin extracted from the sponge Hippospongia sp. with or without HA and CHI nanoparticle encapsulation were assessed and evaluated in vitro. Results showed that IC50 (half maximal inhibitory concentration) of heteronemin toward T24 human bladder cancer cell viability is approximately 0.18 µg/mL. Both plain and HA nanoparticles-encapsulated heteronemin at 0.2 and 0.4 µg/mL significantly reduced T24 cell viability (P<0.001) while HA nanoparticles-encapsulated heteronemin showed weaker viability-inhibitory effects on L929 fibroblasts compared with plain heteronemin at the identical concentrations. HA and CHI nanoparticles-encapsulated heteronemin exhibited significantly stronger inhibitory effects against migration of T24 human bladder cancer cell than those exerted by plain heteronemin at the same concentrations (P<0.001). The flow cytometric results showed that 0.2 µg/mL HA and CHI nanoparticles-encapsulated heteronemin induced higher early apoptosis rate than that induced by plain heteronemin at the same concentration. These results show that HA and CHI nanoparticle encapsulation is able to elevate anti-migratory and apoptosis-inducing effects exerted by heteronemin against bladder cancer cells in vitro. The in vivo anti-bladder cancer effects of the compound with or without HA/CHI nanoparticle encapsulation will be further investigated and examined using murine tumor models. The data obtained from this study will extensively evaluate of the anti-bladder cancer effects of heteronemin as well as HA/CHI-encapsulated heteronemin and pave a way to develop potential bladder cancer treatment.

Keywords: heteronemin, nanoparticles, hyaluronan, chitosan, bladder cancer

Procedia PDF Downloads 445
427 Exploring Marine Bacteria in the Arabian Gulf Region for Antimicrobial Metabolites

Authors: Julie Connelly, Tanvi Toprani, Xin Xie, Dhinoth Kumar Bangarusamy, Kris C. Gunsalus

Abstract:

The overuse of antibiotics worldwide has contributed to the development of multi-drug resistant (MDR) pathogenic bacterial strains. There is an increasing urgency to discover antibiotics to combat MDR pathogens. The microbiome of the Arabian Gulf is a largely unexplored and potentially rich source of novel bioactive compounds. Microbes that inhabit the Abu Dhabi coastal regions adapt to extreme environments with high salinity, hot temperatures, large temperature fluctuations, and acute exposure to solar energy. The microbes native to this region may produce unique metabolites with therapeutic potential as antibiotics and antifungals. We have isolated 200 pure bacterial strains from mangrove sediments, cyanobacterial mats, and coral reefs of the Abu Dhabi region. In this project, we aim to screen the marine bacterial strains to identify antibiotics, in particular undocumented compounds that show activity against existing antibiotic-resistant strains. We have acquired the ESKAPE pathogen panel, which consists of six antibiotic-resistant gram-positive and gram-negative bacterial pathogens that collectively cause most clinical infections. Our initial efforts of the primary screen using colony-picking co-culture assay have identified several candidate marine strains producing potential antibiotic compounds. We will next apply different assays, including disk-diffusion and broth turbidity growth assay, to confirm the results. This will be followed by bioactivity-guided purification and characterization of target compounds from the scaled-up volume of candidate strains, including SPE fraction, HPLC fraction, LC-MS, and NMR. For antimicrobial compounds with unknown structures, our final goal is to investigate their mode of action by identifying the molecular target.

Keywords: marine bacteria, natural products, drug discovery, ESKAPE panel

Procedia PDF Downloads 57
426 Pioglitazone Ameliorates Methotrexate-Induced Renal Endothelial Dysfunction via Amending Detrimental Changes in Antioxidant Profile, Systemic Cytokines and Fas Production

Authors: Sahar M. El-Gowilly, Mai M. Helmy, Hanan M. El-Gowelli

Abstract:

Methotrexate (MTX) is widely used in treatment of cancers and autoimmune diseases. However, nephrotoxicity is one of its most important side effects. The peroxisome proliferator-activated receptor gamma agonist, pioglitazone, is known to exert antiinflammatory and reno-protective effects in various kidney injuries. The purpose of this study was to investigate the potential involvement of endothelial damage in MTX-induced renal injury and to elaborate the possible protective effect of pioglitazone against MTX-induced endothelial impairment. Compared with saline-treated rats, treatment with MTX (7 mg/kg for 3 day) caused significant elevations in serum levels of urea and creatinine, increased renal nitrate/nitrite level and impaired renovascular responsiveness of isolated perfused kidney to endothelium-dependent vasodilations induced by acetylcholine (0.01-2.43 nmol) and isoprenaline (1µmol). These effects were abolished by concurrent treatment with pioglitazone (2.5 mg/kg, for 5 days starting two days before MTX). Alternatively, MTX treatment did not affect endothelium-independent renovascular relaxation induced by sodium nitroprusside (0.001-10 μmole). The possibility that alterations in renal antioxidants, circulating cytokine and apoptotic factor (Fas) levels contributed to MTX-pioglitazone interaction was assessed. Pioglitazone treatment abrogated renal oxidative stress (decreased reduced glutathione and catalase activity and increased malondialdehyde), elevated serum cytokine (interleukin-6, interleukin-10, tumor necrosis factor-alpha and transforming growth factor-beta1) and Fas induced by MTX. Histologically, MTX caused defused tubular cells swelling and vacuolization associated with endothelial damage in renal arterioles. These effects disappeared upon co-treated with pioglitazone. Collectively, pioglitazone abolished MTX-induced endothelium dysfunction and nephrotoxicity via ameliorating oxidative stress and rectifying cytokines and Fas abnormalities caused by MTX.

Keywords: methotrexate, pioglitazone, endothelium, kidney

Procedia PDF Downloads 484
425 Arthroscopic Fixation of Posterior Cruciate Ligament Avulsion Fracture through Posterior Trans Septal Portal Using Button Fixation Device: Mini Tight Rope

Authors: Ratnakar Rao, Subair Khan, Hari Haran

Abstract:

Posterior cruciate ligament (PCL) avulsion fractures is a rare condition and commonly mismanaged.Surgical reattachment has been shown to produce better result compared with conservative management.Only few techniques are reported in arthroscopic fixation of PCL Avulsion Fracture and they are complex.We describe a new technique in fixation of the PCL Avulsion fracture through a posterior trans septal portal using button fixation device (Mini Tight Rope). Eighteen patients with an isolated posterior cruciate ligament avulsion fracture were operated under arthroscopy. Standard Antero Medial Portal and Antero Lateral portals made and additional Postero Medial and Postero Lateral portals made and trans Septal portal established. Avulsion fracture identified, elevated, prepared. Reduction achieved using PCL Tibial guide (Arthrex) and fixation was achieved using Mini Tight Rope,Arthrex (2 buttons with a suture). Reduction confirmed using probe and Image intensifier. Postoperative assessment made clinically and radiologically. 15 patients had good to excellent results with no posterior sag or instability. The range of motion was normal. No complications were recorded per operatively. 2 patients had communition of the fragment while drilling, for one patient it was managed by suturing technique and the second patient PCL Reconstruction was done. One patient had persistent instability with poor outcome. Establishing trans septal portal helps in better visualization of the posterior compartment of the knee. Assessment of the bony fragment, preparation 0f the bone bed andit protects from injury to posterior neurovascular structures. Fixation using the button with suture (Mini Tight Rope) is stable and easily reproducible for PCL Avulsion fracture with single large fragment.

Keywords: PCL avulsion, arthroscopy, transeptal, minitight rope technique

Procedia PDF Downloads 240
424 Copper Chelation by 3-(Bromoacetyl) Coumarin Derivative Induced Apoptosis in Cancer Cells: Influence of Copper Chelation Strategy in Cancer Treatment

Authors: Saman Khan, Imrana Naseem

Abstract:

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

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

Procedia PDF Downloads 272
423 One Health Approach: The Importance of Improving the Identification of Waterborne Bacteria in Austrian Water

Authors: Aurora Gitto, Philipp Proksch

Abstract:

The presence of various microorganisms (bacteria, fungi) in surface water and groundwater represents an important issue for human health worldwide. The matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS) has emerged as a promising and reliable tool for bacteria identification in clinical diagnostic microbiology and environmental strains thanks to an ionization technique that uses a laser energy absorbing matrix to create ions from large molecules with minimal fragmentation. The study aims first to conceptualise and set up library information and create a comprehensive database of MALDI-TOF-MS spectra from environmental water samples. The samples were analysed over a year (2021-2022) using membrane filtration methodology (0.45 μm and 0.22 μm) and then isolated on R2A agar for a period of 5 days and Yeast extract agar growing at 22 °C up to 4 days and 37 °C for 48 hours. The undetected organisms by MALDI-TOF-MS were analysed by PCR and then sequenced. The information obtained by the sequencing was further implemented in the MALDI-TOF-MS library. Among the culturable bacteria, the results show how the incubator temperature affects the growth of some genera instead of others, as demonstrated by Pseudomonas sp., which grows at 22 °C, compared to Bacillus sp., which is abundant at 37 °C. The bacteria community shows a variation in composition also between the media used, as demonstrated with R2A agar which has been defined by a higher presence of organisms not detected compared to YEA. Interesting is the variability of the Genus over one year of sampling and how the seasonality impacts the bacteria community; in fact, in some sampling locations, we observed how the composition changed, moving from winter to spring and summer. In conclusion, the bacteria community in groundwater and river bank filtration represents important information that needs to be added to the library to simplify future water quality analysis but mainly to prevent potential risks to human health.

Keywords: water quality, MALDI-TOF-MS, sequencing, library

Procedia PDF Downloads 64
422 GC-MS Analysis of Bioactive Compounds in the Ethanolic Extract of Nest Material of Mud Wasp, Sceliphron caementarium

Authors: P. Susheela, Mary Rosaline, R. Radha

Abstract:

This research was designed to determine the bioactive compounds present in the nest samples of the mud dauber wasp, Sceliophron caementarium. Insects and insect-based products have been used for the treatment of various ailments from a very long time. It has been found that all over the world including the western societies and the indigenous populations, the usage of insect-based medicine plays an important role in various healing practices and magic rituals. Studies on the therapeutic usage of insects are negligible when compared to plants, the. In the present scenario, it is important to explore bioactive compounds from natural sources rather than depending on synthetic drugs that have adverse effects on human body. Keeping this in view, an attempt was made to analyze and identify bioactive components from the nest sample of the mud dauber wasp, Sceliophron caementarium. The nests of the mud dauber wasp, Sceliophron caementarium were collected from Coimbatore, Tamil Nadu, India. The nest sample was extracted with ethanol for 6-8 hours using Soxhlet apparatus. The final residue was obtained by filtering the extract through Whatman filter paper No.41. The GCMS analysis of the nest sample was performed using Perkin Elmer Elite - 5 capillary column. The resultant compounds were compared with the database of National Institute Standard and Technology (NIST), WILEY8, FAME. The GC-MS analysis of the concentrated ethanol extract revealed the presence of eight constituents like Methylene chloride, Eicosanoic acid, 1, 1’:3’, 1’’-Terphenyl, 5'-Phenyl, Di-N-Decylsulfone, 1, 2-Bis (Trimethylsilyl) Benzene, Androstane-11, 17-Dione, 3-[(Trimethylsilyl) Oxy]-, 17-[O-(Phenylmethyl) O. Most of the identified compounds were reported as having biological activities viz. anti-inflammatory, antibacterial and antifungal properties that can be of pharmaceutical importance and further study of these isolated compounds may prove their medicinal importance in future.

Keywords: Sceliophron caementarium, Gas chromatography-mass spectrometry, ethanol extract, bioactive compounds

Procedia PDF Downloads 270
421 Operational Characteristics of the Road Surface Improvement

Authors: Iuri Salukvadze

Abstract:

Construction takes importance role in the history of mankind, there is not a single thing-product in our lives in which the builder’s work was not to be materialized, because to create all of it requires setting up factories, roads, and bridges, etc. The function of the Republic of Georgia, as part of the connecting Europe-Asia transport corridor, is significantly increased. In the context of transit function a large part of the cargo traffic belongs to motor transport, hence the improvement of motor roads transport infrastructure is rather important and rise the new, increased operational demands for existing as well as new motor roads. Construction of the durable road surface is related to rather large values, but because of high transport-operational properties, such as high-speed, less fuel consumption, less depreciation of tires, etc. If the traffic intensity is high, therefore the reimbursement of expenses occurs rapidly and accordingly is increasing income. If the traffic intensity is relatively small, it is recommended to use lightened structures of road carpet in order to pay for capital investments amounted to no more than normative one. The road carpet is divided into the following basic types: asphaltic concrete and cement concrete. Asphaltic concrete is the most perfect type of road carpet. It is arranged in two or three layers on rigid foundation and will be compacted. Asphaltic concrete is artificial building material, which due stratum will be selected and measured from stone skeleton and sand, interconnected by bitumen and a mixture of mineral powder. Less strictly selected similar material is called as bitumen-mineral mixture. Asphaltic concrete is non-rigid building material and well durable on vertical loadings; it is less resistant to the impact of horizontal forces. The cement concrete is monolithic and durable material, it is well durable the horizontal loads and is less resistant related to vertical loads. The cement concrete consists from strictly selected, measured stone material and sand, the binder is cement. The cement concrete road carpet represents separate slabs of sizes from 3 ÷ 5 op to 6 ÷ 8 meters. The slabs are reinforced by a rather complex system. Between the slabs are arranged seams that are designed for avoiding of additional stresses due temperature fluctuations on the length of slabs. For the joint behavior of separate slabs, they are connected by metal rods. Rods provide the changes in the length of slabs and distribute to the slab vertical forces and bending moments. The foundation layers will be extremely durable, for that is required high-quality stone material, cement, and metal. The qualification work aims to: in order for improvement of traffic conditions on motor roads to prolong operational conditions and improving their characteristics. The work consists from three chapters, 80 pages, 5 tables and 5 figures. In the work are stated general concepts as well as carried out by various companies using modern methods tests and their results. In the chapter III are stated carried by us tests related to this issue and specific examples to improving the operational characteristics.

Keywords: asphalt, cement, cylindrikal sample of asphalt, building

Procedia PDF Downloads 203
420 Nutritional Impact in Patients Who Underwent Sleeve-Type Bariatric Surgery

Authors: Melissa Mattos, Camila Lima, Ibraim Castro, Augusto Carioca, Saulo Magalhães, Paula Freitas, Keciany Oliveira

Abstract:

Obesity is a chronic, multifactorial, relapsing disease that has increased dramatically over the years. Its control is considered a public health issue, and more and more treatments and interventions are being studied to reduce its prevalence. When interventions in lifestyle and the use of drugs do not generate lasting results, bariatric procedures emerge as a resource for obesity control. The main guidelines for the treatment of obesity emphasize the need for pre-procedure and post-procedure nutritional monitoring to avoid nutritional deficiencies that may occur. The individual who undergoes bariatric surgery needs to understand the changes that will be necessary for life in view of the intense anatomical and metabolic changes that result from surgical techniques. To assess the nutritional profile of patients who undergo bariatric surgery, we analyzed data from the medical records of all people who underwent sleeve-type bariatric surgery from January to June 2022 at a clinic in the City of Fortaleza. 38 patients were analyzed, 32 women and 6 men in the pre-surgical period, 6 and 12 months after surgery. The data showed an average weight loss of 24.45% at 6 months and 30.85% at 12 months, with a reduction of 21.32% and 30.41%, respectively, in the fat percentage, also indicating that 13.15% used drugs for weight loss during this period, leading to reflection on the isolated long-term efficacy of bariatric surgery, requiring multidisciplinary follow-up for a change in lifestyle. Only 12 individuals, corresponding to 31.57%, reached eutrophic BMI 12 months after surgery, 20 individuals remained overweight, corresponding to 52.63% of the sample, and 6 individuals (15.78%) remained in the BMI obese class I. As for body composition, there was a 52.39% reduction in fat mass and a 12.82% reduction in muscle mass, and 21% of individuals underwent cholecystectomy. Sleeve-type bariatric surgery promoted significant weight loss after 1 year of the procedure, with a reduction in body fat percentage and fat mass. Most patients were still overweight and had a significant reduction in muscle mass.

Keywords: bariatric surgery, sleeve gastrectomy, obesity, sleeve

Procedia PDF Downloads 49
419 Chemical Composition and Antifungal Activity of Selected Essential Oils against Toxigenic Fungi Associated with Maize (Zea mays L.)

Authors: Birhane Atnafu, Chemeda Abedeta Garbaba, Fikre Lemessa, Abdi Mohammed, Alemayehu Chala

Abstract:

Essential oil is a bio-pesticide plant product used as an alternative to pesticides in managing plant pests, including fungal pathogens. Thus, the current study aims to investigate the chemical composition and antifungal activities of essential oils (EO) extracted from three aromatic plants i.e., Thymus vulgaris, Coriandrum sativum, and Cymbopogon martini. The leaf parts of those selected plants were collected from the Jimma area and their essential oil was extracted by hydro-distillation method in a Clevenger apparatus. The chemical composition of selected plant essential oil was analyzed by using Gas chromatography-mass spectrometry (GC/MS) and their inhibitory effects were tested in vitro on toxigenic fungi isolated from maize kernel. Chemical analysis results revealed the presence of 32 compounds in C. sativum with Hexanedioic acid, bis (2-ethylhexyl) ester (46. 9%), 2-Decenal, (E)- (12.6), and linalool (8.3%) being the dominant ones. T. vulgaris essential oils constituted 25 compounds, of which thymol (34.4%), o-cymene (17.5%), and Gamma-Terpinene (16.8%) were the major components. Twenty-five compounds were detected in C. martinii of which geraniol (51.4%), Geranyl acetate (14.5%), and Trans – ß-Ocimene (11.7%) were dominant. The EOs of the tested plants had very high antifungal activity (up to 100% efficacy) against Aspergillus flavus, Aspergillus niger, Fusarium graminearum and Fusarium verticillioides in vitro and on maize grains. The antifungal activities of these essential oils were dependent on the major components such as thymol, hexanedioic acid, bis (2-ethylhexyl) ester, and geraniol. The study affirmed the potential of these essential oils controlling as bio-fungicides to manage the effects of potentially toxigenic fungi associated with maize under post-harvest stages. This can reduce the consequences of the health impacts of the mold and toxigenic compounds produced in maize.

Keywords: bio-activity, bio-pesticides, maize, mycotoxin

Procedia PDF Downloads 49
418 A Review of Farmer Participation in Information and Communication Technology through Mobile Banking and Mobile Marketing in Rural Agricultural Systems

Authors: J. Cadby, K. Miyazawa

Abstract:

Information and Communication Technology (ICT) has been widely adopted into the agricultural landscape with advancements of mobile connectivity and data accessibility. In developed nations, mobile-technology is well integrated into marketing transactions, and also plays a crucial role in making data-driven decisions on-farm. In developing nations, mobile banking and access to agricultural extension services allow for informed decision-making and smoother transactions. In addition, the availability of updated and readily available market and climate data provides a negotiation platform, reducing economic risks for farmers worldwide. The total usage of mobile technology has risen over the past 20 years, and almost three-quarters of the world’s population subscribes to mobile technology. This study reviewed mobile technology integration into agricultural systems in developing and developed nations. Data from secondary sources were collected and investigated. The objectives of the study include a review of the success of mobile banking transactions in developing nations, and a review of application and SMS based services for direct marketing in both developed and developing nations. Rural farmers in developing countries with access to diverse m-banking options experienced increased access to farm investment resources with the use of mobile banking technology. Rural farmers involved in perishable crop production were also more likely to benefit from mobile platform sales participation. ICT programs reached through mobile application and SMS increased access to agricultural extension materials and marketing tools for demographics that faced literacy-challenges and isolated markets. As mobile technology becomes more ubiquitous in the global agricultural system, training and market opportunities to facilitate mobile usage in developing agricultural systems are necessary. Digital skills training programs are necessary in order to improve equal global adoption of ICT in agriculture.

Keywords: market participation, mobile banking, mobile technology, rural farming

Procedia PDF Downloads 229
417 Political Communication in Twitter Interactions between Government, News Media and Citizens in Mexico

Authors: Jorge Cortés, Alejandra Martínez, Carlos Pérez, Anaid Simón

Abstract:

The presence of government, news media, and general citizenry in social media allows considering interactions between them as a form of political communication (i.e. the public exchange of contradictory discourses about politics). Twitter’s asymmetrical following model (users can follow, mention or reply to other users that do not follow them) could foster alternative democratic practices and have an impact on Mexican political culture, which has been marked by a lack of direct communication channels between these actors. The research aim is to assess Twitter’s role in political communication practices through the analysis of interaction dynamics between government, news media, and citizens by extracting and visualizing data from Twitter’s API to observe general behavior patterns. The hypothesis is that regardless the fact that Twitter’s features enable direct and horizontal interactions between actors, users repeat traditional dynamics of interaction, without taking full advantage of the possibilities of this medium. Through an interdisciplinary team including Communication Strategies, Information Design, and Interaction Systems, the activity on Twitter generated by the controversy over the presence of Uber in Mexico City was analysed; an issue of public interest, involving aspects such as public opinion, economic interests and a legal dimension. This research includes techniques from social network analysis (SNA), a methodological approach focused on the comprehension of the relationships between actors through the visual representation and measurement of network characteristics. The analysis of the Uber event comprised data extraction, data categorization, corpus construction, corpus visualization and analysis. On the recovery stage TAGS, a Google Sheet template, was used to extract tweets that included the hashtags #UberSeQueda and #UberSeVa, posts containing the string Uber and tweets directed to @uber_mx. Using scripts written in Python, the data was filtered, discarding tweets with no interaction (replies, retweets or mentions) and locations outside of México. Considerations regarding bots and the omission of anecdotal posts were also taken into account. The utility of graphs to observe interactions of political communication in general was confirmed by the analysis of visualizations generated with programs such as Gephi and NodeXL. However, some aspects require improvements to obtain more useful visual representations for this type of research. For example, link¬crossings complicates following the direction of an interaction forcing users to manipulate the graph to see it clearly. It was concluded that some practices prevalent in political communication in Mexico are replicated in Twitter. Media actors tend to group together instead of interact with others. The political system tends to tweet as an advertising strategy rather than to generate dialogue. However, some actors were identified as bridges establishing communication between the three spheres, generating a more democratic exercise and taking advantage of Twitter’s possibilities. Although interactions in Twitter could become an alternative to political communication, this potential depends on the intentions of the participants and to what extent they are aiming for collaborative and direct communications. Further research is needed to get a deeper understanding on the political behavior of Twitter users and the possibilities of SNA for its analysis.

Keywords: interaction, political communication, social network analysis, Twitter

Procedia PDF Downloads 203
416 Potential Application of Thyme (Thymus vulgaris L.) Essential Oil as Antibacterial Drug in Aromatherapy

Authors: Ferhat Mohamed Amine, Boukhatem Mohamed Nadjib, Chemat Farid

Abstract:

The Lamiaceae family is widely spread in Algeria. Due to the application of Thymus species growing wild in Algeria as a culinary herb and in folk medicine, the purpose of the present work was to evaluate antimicrobial activities of their essential oils and relate them with their chemical composition, for further application in food and pharmaceutical industries as natural valuable products. The extraction of the Thymus vulgaris L. essential oil (TVEO) was obtained by steam distillation. Chemical composition of the TVEO was determined by Gas Chromatography. A total of thirteen compounds were identified. Carvacrol (83.8%) was the major component, followed by cymene (8.15%) and terpinene (4.96%). Antibacterial action of the TVEO against 23 clinically isolated bacterial strains was determined by using agar disc diffusion and vapour diffusion methods at different doses. By disc diffusion method, TVEO showed potent antimicrobial activity against gram-positive bacteria more than gram-negative strains and antibiotic discs. The Diameter of Inhibition Zone (DIZ) varied from 25 to 60 mm for S. aureus, B. subtilisand E. coli. However, the results obtained by both agar diffusion and vapour diffusion methods were different. Significantly higher antibacterial effect was observed in the vapour phase at lower doses. S. aureus and B. subtilis were the most susceptible strains to the oil vapour. Therefore, smaller doses of EO in the vapour phase can be inhibitory to pathogenic bacteria. There is growing evidence that TVEO in vapour phase are effective antiseptic systems and appears worthy to be considered for practical uses in the treatment of human infections oras air decontaminants in hospital. TVEO has considerable antibacterial activity deserving further investigation for clinical applications. Also whilst the mode of action remains mainly undetermined, this experimental approach will need to continue.

Keywords: antimicrobial drugs, carvacrol, disc diffusion, Thymus vulgaris, vapour diffusion

Procedia PDF Downloads 353
415 Comparison Study of 70% Ethanol Effect on Direct and Retrival Culture of Contaminated Umblical Cord Tissue for Expansion of Mesenchymal Stem Cells

Authors: Ganeshkumar, Ashika, Valavan, Ramesh, Thangam, Chirayu

Abstract:

MSCs are found in much higher concentration in the Wharton’s jelly compared to the umbilical cord blood, which is a rich source of hematopoietic stem cells. Umbilical cord tissue is collected at the time of birth; it is processed and stored in liquid nitrogen for future therapeutical purpose. The source of contamination might be either from vaginal tract of mother or from hospital environment or from personal handling during cord tissue sample collection. If the sample were contaminated, decontamination procedure will be done with 70% ethanol (1 minute) in order to avoid sample rejection. Ethanol is effective against a wide range of bacteria, protozoa and fungi and has low toxicity to humans. Among the 1954 samples taken for the study, 24 samples were found to be contaminated with microorganism. The organisms isolated from the positive samples were found to be E. coli, Stenotrophomonas maltophilia, Pseudomonas aueroginosa, Enterococcus fecalis, Acinetobacter bowmani, Staphylococcus epidermidis, Enterobacter cloacae, and Proteus mirabilis. Among these organisms 70% ethanol successfully eliminated E. coli, Enterococcus fecalis, Acinetobacter bowmani, Staphylococcus epidermidis, and Proteus mirabilis. 70% ethanol was unsuccessful in eliminating Stenotrophomonas maltophilia, Pseudomonas aueroginosa, and Enterobacter cloacae. Stenotrophomonas maltophilia and Pseudomonas aueroginosa have the ability to form biofilm that make them resistant to alcohol. Biofilm act as protective layer for bacteria and which protects them from host defense and antibiotic wash. Finally it was found 70% ethanol wash saved 58.3% cord tissue samples from rejection and it is ineffective against 41% of the samples. The contamination rate can be reduced by maintaining proper aseptic techniques during sample collection and processing.

Keywords: umblical cord tissue, decontamination, 70% ethanol effectiveness, contamination

Procedia PDF Downloads 335
414 Localisation of Fluorescently Labelled Drug-Free Phospholipid Vesicles to the Cartilage Surface of Rat Synovial Joints

Authors: Sam Yurdakul, Nick Baverstock, Jim Mills

Abstract:

TDT 064 (FLEXISEQ®) is a drug-free gel used to treat osteoarthritis (OA)-associated pain and joint stiffness. It contains ultra-deformable phospholipid Sequessome™ vesicles, which can pass through the skin barrier intact. In six randomized OA studies, topical TDT 064 was well tolerated and improved joint pain, physical function and stiffness. In the largest study, these TDT 064-mediated effects were statistically significantly greater than oral placebo and equivalent to celecoxib. To understand the therapeutic effects of TDT 064, we investigated the localisation of the drug-free vesicles within rat synovial joints. TDT 064 containing DiO-labelled Sequessome™ vesicles was applied to the knees of four 6-week-old CD® hairless rats (10 mg/kg/ joint), 2–3 times/day, for 3 days (representing the recommended clinical dose). Eighteen hours later, the animals and one untreated control were sacrificed, and the knee joints isolated, flash frozen and embedded in Acrytol Mounting Media™. Approximately 15 sections (10 µm) from each joint were analysed by fluorescence microscopy. To investigate whether the localisation of DiO fluorescence was associated with intact vesicles, an anti-PEG monoclonal antibody (mAb) was used to detect Tween, a constituent of Sequessome™ vesicles. Sections were visualized at 484 nm (DiO) and 647 nm (anti-PEG mAb) and analysed using inForm 1.4 (Perkin Elmer, Inc.). Significant fluorescence was observed at 484 nm in sections from TDT 064-treated animals. No non-specific fluorescence was observed in control sections. Fluorescence was detected as discrete vesicles on the cartilage surfaces, inside the cartilaginous matrix and within the synovial space. The number of DiO-labelled vesicles in multiple fields of view was consistent and >100 in sections from four different treated knees. DiO and anti-PEG mAb co-localised within the collagenous tissues in four different joint sections. Under higher magnification (40x), vesicles were seen in the intercellular spaces of the synovial joint tissue, but no fluorescence was seen inside cells. These data suggest that the phospholipid vesicles in TDT 064 localize at the surface of the joint cartilage; these vesicles may therefore be supplementing the phospholipid deficiency reported in OA and acting as a biolubricant within the synovial joint.

Keywords: joint pain, osteoarthritis, phospholipid vesicles, TDT 064

Procedia PDF Downloads 426
413 To Smile or Not to Smile: How Engendered Facial Cues affect Hiring Decisions

Authors: Sabrina S. W. Chan, Emily Schwartzman, Nicholas O. Rule

Abstract:

Past literature showed mixed findings on how smiling affects a person’s chance of getting hired. On one hand, smiling suggests enthusiasm, cooperativeness, and enthusiasm, which can elicit positive impressions. On the other hand, smiling can suggest weaker professionalism or a filler to hide nervousness, which can lower a candidate’s perceived competence. Emotion expressions can also be perceived differently depending on the person’s gender and can activate certain gender stereotypes. Women especially face a double bind with respect to hiring decisions and smiling. Because women are socially expected to smile more, those who do not smile will be considered stereotype incongruent. This becomes a noisy signal to employers and may lower their chance of being hired. However, women’s smiling as a formality may also be an obstacle. They are more likely to put on fake smiles; but if they do, they are also likely to be perceived as inauthentic and over-expressive. This paper sought to investigate how smiling affects hiring decisions, and whether this relationship is moderated by gender. In Study 1, participants were shown a series of smiling and emotionally neutral face images, incorporated into fabricated LinkedIn profiles. Participants were asked to rate how hireable they thought that candidate was. Results showed that participants rated smiling candidates as more hireable than nonsmiling candidates, and that there was no difference in gender. Moreover, individuals who did not study business were more biased in their perceptions than those who did. Since results showed a trending favoritism over female targets, in suspect of desirability bias, a second study was conducted to collect implicit measures behind the decision-making process. In Study 2, a mouse-tracking design was adopted to explore whether participants’ implicit attitudes were different from their explicit responses on hiring. Participants asked to respond whether they would offer an interview to a candidate. Findings from Study 1 was replicated in that smiling candidates received more offers than neutral-faced candidates. Results also showed that female candidates received significantly more offers than male candidates but was associated with higher attractiveness ratings. There were no significant findings in reaction time or change of decisions. However, stronger hesitation was detected for responses made towards neutral targets when participants perceived the given position as masculine, implying a conscious attempt of making situational judgments (e.g., considering candidate’s personality and job fit) to override automatic processing (evaluations based on attractiveness). Future studies would look at how these findings differ for positions which are stereotypically masculine (e.g., surgeons) and stereotypically feminine (e.g., kindergarten teachers). Current findings have strong implications for developing bias-free hiring policies in workplace, especially for organizations who maintain online/hybrid working arrangements in the post-pandemic era. This also bridges the literature gap between face perception and gender discrimination, highlighting how engendered facial cues can affect individual’s career development and organization’s success in diversity and inclusion.

Keywords: engendered facial cues, face perception, gender stereotypes, hiring decisions, smiling, workplace discrimination

Procedia PDF Downloads 107
412 Revealing the Genome Based Biosynthetic Potential of a Streptomyces sp. Isolate BR123 Presenting Broad Spectrum Antimicrobial Activities

Authors: Neelma Ashraf

Abstract:

Actinomycetes, particularly genus Streptomyces is of great importance due to their role in the discovery of new natural products, particularly antimicrobial secondary metabolites in the medicinal science and biotechnology industry. Different Streptomyces strains were isolated from Helianthus annuus plants and tested for antibacterial and antifungal activities. The most promising five strains were chosen for further investigation, and growth conditions for antibiotic synthesis were optimised. The supernatants were extracted in different solvents, and the extracted products were analyzed using liquid chromatography-mass spectrometry (LC-MS) and biological testing. From one of the potent strains Streptomyces globusus sp. BR123, a compound lavendamycin was identified using these analytical techniques. In addition, this potent strain also produces a strong antifungal polyene compound with a quasimolecular ion of 2072. Streptomyces sp. BR123 was genome sequenced because of its promising antimicrobial potential in order to identify the gene cluster responsible for analyzed compound “lavendamycin”. The genome analysis yielded candidate genes responsible for the production of this potent compound. The genome sequence of 8.15 Mb of Streptomyces sp. isolate BR123 with a GC content of 72.63% and 8103 protein coding genes was attained. Many antimicrobial, antiparasitic, and anticancerous compounds were detected through multiple biosynthetic gene clusters predicted by in-Silico analysis. Though, the novelty of metabolites was determined through the insignificant resemblance with known biosynthetic gene clusters. The current study gives insight into the bioactive potential of Streptomyces sp. isolate BR123 with respect to the synthesis of bioactive secondary metabolites through genomic and spectrometric analysis. Moreover, the comparative genome study revealed the connection of isolate BR123 with other Streptomyces strains, which could expand the knowledge of this genus and the mechanism involved in the discovery of new antimicrobial metabolites.

Keywords: streptomyces, secondary metabolites, genome, biosynthetic gene clusters, high performance liquid chromatography, mass spectrometry

Procedia PDF Downloads 55
411 High Level Expression of Fluorinase in Escherichia Coli and Pichia Pastoris

Authors: Lee A. Browne, K. Rumbold

Abstract:

The first fluorinating enzyme, 5'-fluoro-5'-deoxyadenosine synthase (fluorinase) was isolated from the soil bacterium Streptomyces cattleya. Such an enzyme, with the ability to catalyze a C-F bond, presents great potential as a biocatalyst. Naturally fluorinated compounds are extremely rare in nature. As a result, the number of fluorinases identified remains relatively few. The field of fluorination is almost completely synthetic. However, with the increasing demand for fluorinated organic compounds of commercial value in the agrochemical, pharmaceutical and materials industries, it has become necessary to utilize biologically based methods such as biocatalysts. A key step in this crucial process is the large-scale production of the fluorinase enzyme in considerable quantities for industrial applications. Thus, this study aimed to optimize expression of the fluorinase enzyme in both prokaryotic and eukaryotic expression systems in order to obtain high protein yields. The fluorinase gene was cloned into the pET 41b(+) and pPinkα-HC vectors and used to transform the expression hosts, E.coli BL21(DE3) and Pichia pastoris (PichiaPink™ strains) respectively. Expression trials were conducted to select optimal conditions for expression in both expression systems. Fluorinase catalyses a reaction between S-adenosyl-L-Methionine (SAM) and fluoride ion to produce 5'-fluorodeoxyadenosine (5'FDA) and L-Methionine. The activity of the enzyme was determined using HPLC by measuring the product of the reaction 5'FDA. A gradient mobile phase of 95:5 v/v 50mM potassium phosphate buffer to a final mobile phase containing 80:20 v/v 50mM potassium phosphate buffer and acetonitrile were used. This resulted in the complete separation of SAM and 5’-FDA which eluted at 1.3 minutes and 3.4 minutes respectively. This proved that the fluorinase enzyme was active. Optimising expression of the fluorinase enzyme was successful in both E.coli and PichiaPink™ where high expression levels in both expression systems were achieved. Protein production will be scaled up in PichiaPink™ using fermentation to achieve large-scale protein production. High level expression of protein is essential in biocatalysis for the availability of enzymes for industrial applications.

Keywords: biocatalyst, expression, fluorinase, PichiaPink™

Procedia PDF Downloads 534
410 A Systematic Review with Meta-Analyses Investigating the Association between Binge Eating and Poor Weight Loss Outcomes in People with Obesity

Authors: Isabella Lobo Sasaoka, Felipe Q. da Luz, Zubeyir Salis, Phillipa Hay, Tamiris Gaeta, Paula Costa Teixeira, Táki Cordás, Amanda Sainsbury

Abstract:

Background: A significant number of people with obesity that seek weight loss treatments experience binge eating episodes. Nonetheless, it is unknown whether binge eating episodes can hinder weight loss outcomes. Objective: To compare weight change in people with or without binge eating submitted to bariatric surgery, pharmacotherapy, nutritional orientation, and/or psychological therapies. Method: We conducted a systematic review with meta-analyses by searching studies in PubMed, American Psychological Association (APA), and Embase. Results: Thirty-four studies were included in our systematic review, and 17 studies were included in the meta-analyses. Overall, we found no significant difference in weight loss between people with or without binge eating submitted to any type of weight loss treatment. Additionally, we found no statistically significant differences in body weight between people with or without binge eating at short and long follow-up assessments following any type of weight loss treatment. We also examined changes in body weight in people with or without binge eating in three additional meta-analyses categorized by the type of weight loss treatment (i.e., behavioural and/or nutritional interventions; bariatric surgery; pharmacotherapy isolated or combined with behavior interventions) and found no difference in weight loss. Eleven out of the 17 studies that were assessed qualitatively (i.e., not included in meta-analyses) did not show differences in weight loss in people with or without binge eating submitted to any type of weight loss treatment. Conclusion: This systematic review with meta-analyses showed no difference in weight loss in people with or without binge eating submitted to a variety of weight loss treatments. Nonetheless, specialized therapies can be required to address eating disorder psychopathology and recurrent binge eating in people with obesity that seek weight loss.

Keywords: obesity, binge eating, weight loss, systematic review, meta-analysis

Procedia PDF Downloads 131
409 Comparative Parametric Analysis on the Dynamic Response of Fibre Composite Beams with Debonding

Authors: Indunil Jayatilake, Warna Karunasena

Abstract:

Fiber Reinforced Polymer (FRP) composites enjoy an array of applications ranging from aerospace, marine and military to automobile, recreational and civil industry due to their outstanding properties. A structural glass fiber reinforced polymer (GFRP) composite sandwich panel made from E-glass fiber skin and a modified phenolic core has been manufactured in Australia for civil engineering applications. One of the major mechanisms of damage in FRP composites is skin-core debonding. The presence of debonding is of great concern not only because it severely affects the strength but also it modifies the dynamic characteristics of the structure, including natural frequency and vibration modes. This paper deals with the investigation of the dynamic characteristics of a GFRP beam with single and multiple debonding by finite element based numerical simulations and analyses using the STRAND7 finite element (FE) software package. Three-dimensional computer models have been developed and numerical simulations were done to assess the dynamic behavior. The FE model developed has been validated with published experimental, analytical and numerical results for fully bonded as well as debonded beams. A comparative analysis is carried out based on a comprehensive parametric investigation. It is observed that the reduction in natural frequency is more affected by single debonding than the equally sized multiple debonding regions located symmetrically to the single debonding position. Thus it is revealed that a large single debonding area leads to more damage in terms of natural frequency reduction than isolated small debonding zones of equivalent area, appearing in the GFRP beam. Furthermore, the extents of natural frequency shifts seem mode-dependent and do not seem to have a monotonous trend of increasing with the mode numbers.

Keywords: debonding, dynamic response, finite element modelling, novel FRP beams

Procedia PDF Downloads 105
408 Studies on Knockdown Resistance Mutations in Aedes aegypti and Aedes albopictus in India

Authors: Neera Kapoor

Abstract:

Background: Knockdown Resistance (KDR) is one of the mechanisms of insecticide resistance in insects caused by the reduced target site sensitivity i.e. voltage gated sodium channel (VGSC) rendering it less sensitive to the toxic effects of DDT and pyrethroids. In this study, we evaluated insecticide susceptibility and its underlying KDR mechanism in eight Ae. aegypti and five Ae. albopictus field populations. Methodology: Field population was collected from four different geographical regions of India covering 18 districts of ten states. For genotyping of twelve KDR alleles in Ae. aegypti field populations, three PCR based assays were used; with DNA sequencing; ASPCR; PCR-RFLP. Genomic DNA was isolated, and three partial domains (II, III, and IV) of VGSC were amplified and sequenced. Results: Molecular screening for common KDR mutations, revealed the presence of five mutations viz. S989P, V1016G, T1520I, F1534C/L. Two novel mutations were observed, first at T1520 (ACC) residue where a C > T substitution at the second position of codon results in amino acid change to Isoleucine (ATC). Second mutation was an alternative point mutation at F1534 (TTC) residue where a substitution of T > C at the first position of codon results in an amino acid change to Leucine (CTC). ASPCRs were not accurate, so three PCR-RFLP assays were developed for genotyping of five KDR alleles in Ae. aegypti; viz. T1520I, F1534C/L. Representative samples of all genotypes (n=200) were sequenced to validate the newly developed PCR based assays for Ae. aegypti. Genotyping results showed that 989P is linked to 1016G and novel mutation 1520I was always found with 1534C allele. Conclusion: Present study confirmed the presence of DDT and pyrethroid resistance among Ae. aegypti populations in India and for the first time reported KDR mutations in this species from India including two novel mutations. Results of present study lead us to infer that, at least five KDR mutations (S989P, V1016G, T1530I, F1534C, and F1534L) can be seen as a potential marker for DDT/pyrethroid resistance.

Keywords: F1534C, F1534L, S989P, T1530I, V1016G

Procedia PDF Downloads 172
407 Effect of Madecassoside on the Antioxidant Status of Streptozotocin-Nicotinamide Induced Diabetes in Sprague-Dawley Rats

Authors: C. Mayuren, C. K. Paul Wang, K. Purushotham, C. Dinesh Kumar

Abstract:

Diabetes Mellitus (DM) is one of the most common non-communicable diseases globally. Although significant advances have led to better understanding of the condition and the development of effective therapies and preventive strategies, the pathway to cure remains elusive and DM prevails as a serious medical challenge in the 21st century. Oxidative stress has been suggested to contribute to the progression and pathophysiological conditions of diabetes. Madecassoside (MA) a major pentacyclic triterpenoid, has been demonstrated to possess various biological activities. However, no attempt has been made to study the antioxidant activity in diabetic rats. Therefore, the present study is aimed to evaluate the antioxidant effect of MA on streptozotocin-nicotinamide induced type-2 diabetes in Sprague-Dawley rats. The study protocol was approved by the institutional ethical committee prior to the conduct of research. Adult male Sprague-Dawley rats weighing 250-300 g were used in the study. The animals were rendered diabetic with a single intraperitoneal dose of streptozotocin (65 mg/kg) and nicotinamide (110 mg/kg). The diabetic animals after a stabilisation period of 14 days received various treatments (Madecassoside 50 mg/kg; Glimepiride 2.5 mg/kg) suspended in 0.5% carboxymethyl cellulose orally, for a period of 28 days. The animals fasted overnight after the last treatment were sacrificed and the pancreas, liver and kidneys were isolated. The weighted quantity of the samples of various treatments were homogenised in ice-cold condition and were subjected to lipid peroxidation, catalase and superoxide dismutase assay. The data’s obtained were subjected to statistical analysis. Diabetic rats showed significant increase in lipid peroxidation and decrease in enzymatic antioxidant levels. All the treated groups had significantly higher SOD, CAT and reduced LPO activity in the pancreas, liver and kidney. Results suggest madecassoside to have potential antioxidant effect against the diabetic model. However further investigations are necessary to study the mechanism at the cellular level.

Keywords: antioxidant, diabetes, madecassoside, nicotinamide, streptozotocin

Procedia PDF Downloads 364
406 The Antibacterial and Anticancer Activity of Marine Actinomycete Strain HP411 Isolated in the Northern Coast of Vietnam

Authors: Huyen T. Pham, Nhue P. Nguyen, Tien Q. Phi, Phuong T. Dang, Hy G. Le

Abstract:

Since the marine environmental conditions are extremely different from the other ones, so that marine actinomycetes might produce novel bioactive compounds. Therefore, actinomycete strains were screened from marine water and sediment samples collected from the coastal areas of Northern Vietnam. Ninety-nine actinomycete strains were obtained on starch-casein agar media by dilution technique, only seven strains, named HP112, HP12, HP411, HPN11, HP 11, HPT13 and HPX12, showed significant antibacterial activity against both gram-positive and gram-negative bacteria (Bacillus subtilis ATCC 6633, Staphylococcus epidemidis ATCC 12228, Escherichia coli ATCC 11105). Further studies were carried out with the most active HP411strain against Candida albicans ATCC 10231. This strain could grow rapidly on starch casein agar and other media with high salt containing 7-10% NaCl at 28-30oC. Spore-chain of HP411 showed an elongated and circular shape with 10 to 30 spores/chain. Identification of the strain was carried out by employing the taxonomical studies including the 16S rRNA sequence. Based on phylogenetic and phenotypic evidence it is proposed that HP411 to be belongs to species Streptomyces variabilis. The potent of the crude extract of fermentation broth of HP411that are effective against wide range of pathogens: both gram-positive, gram-negative and fungi. Further studies revealed that the crude extract HP411 could obtain the anticancer activity for cancer cell lines: Hep-G2 (liver cancer cell line); RD (cardiac and skeletal muscle letters cell line); FL (membrane of the uterus cancer cell line). However, the actinomycetes from marine ecosystem will be useful for the discovery of new drugs in the furture.

Keywords: marine actinomycetes, antibacterial, anticancer, Streptomyces variabilis

Procedia PDF Downloads 403
405 Stereological Evaluation of Liver of Rabbit Fetuses After Transplantation of Human Wharton’s Jelly-Derived Mesenchymal Stromal/Stem Cells

Authors: Zahra Khodabandeh, Leila Rezaeian, Mohammad Amin Edalatmanesh, Asghar Mogheiseh, Nader Tanideh, Mehdi Dianatpour, Shahrokh Zare, Hossein Bordbar, Neda Baghban, Amin Tamadon

Abstract:

Background: In-utero xenotransplantation of stem cells in abnormal fetuses effectively treats several genetic illnesses. Objective: The current research aimed to evaluate structural and morphological alterations in the liver of rabbit fetuses following xenotransplantation of human Wharton’s jelly-derived mesenchymal stromal cells (hWJ-MSCs) using a stereological technique. Methods: hWJ-MSCs were isolated from the human umbilical cord, and their authenticity was established by flow cytometry and differentiation. At gestational day 14, the rabbits were anesthetized, and hWJ-MSCs were injected into the uteri of 24 fetuses. Twenty-two fetuses were born successfully. Ten rabbit liver specimens were prepared from injected fetuses, including eight rabbits on day three following birth and two rabbits on the 21st post-natal day. The non-injected fetuses were considered positive controls. The livers of the control and hWJ-MSCs-treated rabbits were fixed, processed, stained, and examined through stereological approaches. Results: In the hWJ-MSCs-treated group, the mean liver weight and volume increased by 42% and 78% compared to the control group. The total volume of the hepatocytes increased by 63% and that of sinusoids by threefold in the treated rabbits. The total volume of the central veins increased by 70%. The total number corresponding to hepatocytes in the experimental group increased by 112% compared to the rabbits in the control. The total volume of the hepatocyte nuclei in the experimental group increased by 117% compared to the rabbits in the control. Conclusion: After xenotransplantation of human MSCs, host tissue microenvironments (here, the rabbit liver) were altered, and these included quantitative factors corresponding to the liver tissue and hepatocyte morphometric indices.

Keywords: xenotransplantation, mesenchymal stromal, stem cell, Wharton ‘s jelly, liver

Procedia PDF Downloads 69