Search results for: in vitro toxicity
Commenced in January 2007
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Edition: International
Paper Count: 2256

Search results for: in vitro toxicity

636 Screening of Potential Cytotoxic Activities of Some Medicinal Plants of Saudi Arabia

Authors: Syed Farooq Adil, Merajuddinkhan, Mujeeb Khan, Hamad Z. Alkhathlan

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Phytochemicals from plant extracts belong to an important source of natural products which have demonstrated excellent cytotoxic activities. However, plants of different origins exhibit diverse chemical compositions and bioactivities. Therefore, the discovery of plants based new anticancer agents from different parts of the world is always challenging. In this study, methanolic extracts of different parts of 11 plants from Saudi Arabia have been tested in vitro for their anticancer potential on human liver cancer cell line (HepG2). Particularly, for this study, plants from Asteraceae, Resedaceae, and Polygonaceae families were chosen on the basis of locally available ethnobotanical data and their medicinal properties. Among 12 tested extract samples, three samples obtained from Artemisia monosperma stem, Ochradenus baccatus aerial parts, and Pulicaria glutinosa stem have demonstrated interesting cytotoxic activities with a cell viability of 29.3%, 28.4% and 24.2%, respectively. Whereas, four plant extracts including Calendula arvensis aerial parts, Scorzonera musilii whole plant, A. monosperma leaves show moderate anticancer properties bearing a cell viability ranging from 11.9 to 16.7%. The remaining extracts have shown poor cytotoxic activities. Subsequently, GC-MS analysis of methanolic extracts of the four most active plants extracts such as C. comosum, O. baccatus, P. glutinosa and A. monosperma detected the presence of 41 phytomolecules. Among which 3-(4-hydroxyphenyl) propionitrile (1), 8,11-octadecadiynoic acid methyl ester (2), 6,7-dimethoxycoumarin (3), and 1-(2-hydroxyphenyl) ethenone (4) were found to be the lead compounds of C. comosum, O. baccatus P. glutinosa and A. monosperma, respectively.

Keywords: medicinal plants, asteraceae, polygonaceae, hepg2

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635 The Effect of a Weed-Killer Sulfonylurea on Durum Wheat (Triticum Durum Desf)

Authors: L. Meksem Amara, M. Ferfar, N. Meksem, M. R. Djebar

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The wheat is the cereal the most consumed in the world. In Algeria, the production of this cereal covers only 20 in 25 % of the needs for the country, the rest being imported. To improve the efficiency and the productivity of the durum wheat, the farmers turn to the use of pesticides: weed-killers, fungicides and insecticides. However this use often entrains losses of products more at least important contaminating the environment and all the food chain. Weed-killers are substances developed to control or destroy plants considered unwanted. That they are natural or produced by the human being (molecule of synthesis), the absorption and the metabolization of weed-killers by plants cause the death of these plants. In this work, we set as goal the evaluation of the effect of a weed-killer sulfonylurea, the CossackOD with various concentrations (0, 2, 4 and 9 µg) on variety of Triticum durum: Cirta. We evaluated the plant growth by measuring the leaves and root length, compared with the witness as well as the content of proline and analyze the level of one of the antioxydative enzymes: catalase, after 14 days of treatment. Sulfonylurea is foliar and root weed-killers inhibiting the acetolactate synthase: a vegetable enzyme essential to the development of the plant. This inhibition causes the ruling of the growth then the death. The obtained results show a diminution of the average length of leaves and roots this can be explained by the fact that the ALS inhibitors are more active in the young and increasing regions of the plant, what inhibits the cellular division and talks a limitation of the foliar and root’s growth. We also recorded a highly significant increase in the proline levels and a stimulation of the catalase activity. As a response to increasing the herbicide concentrations a particular increases in antioxidative mechanisms in wheat cultivar Cirta suggest that the high sensitivity of Cirta to this sulfonylurea herbicide is related to the enhanced production and oxidative damage of reactive oxygen species.

Keywords: sulfonylurea, triticum durum, oxydative stress, toxicity

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634 Antiprotozoal Activity against Entamoeba histolytica of Flavonoids Isolated from Lippia graveolens Kunth

Authors: Ramiro Quintanilla-Licea, Isvar K. Angeles-Hernandez, Javier Vargas-Villarreal

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Amebiasis caused by Entamoeba histolytica, associated with high morbidity and mortality, is currently a significant public health problem worldwide, especially in developing countries. In the world, around 50 million people develop this illness, and up to 100,000 deaths occur annually. Due to the side-effects and the resistance that pathogenic protozoa show against common antiparasitic drugs (e.g., metronidazole), growing attention has been paid to plants used in traditional medicine around the world to find new antiprotozoal agents. In this study is reported about the isolation and structure elucidation of antiamoebic compounds occurring in Lippia graveolens Kunth (Mexican oregano). The work-up of the methanol extract of L. graveolens afforded the known flavonoids pinocembrin (1), sakuranetin (2), cirsimaritin (3) and naringenin (4) by bioguided isolation using several chromatographic techniques. Structural elucidation of the isolated compounds was based on spectroscopic/spectrometric analyses (IR; 1H- and 13C-NMR; MS) and comparison with literature data. These compounds showed significant antiprotozoal activity against Entamoeba histolytica trophozoites using in vitro tests (positive control metronidazole IC50 0.205 µg/mL). The antiprotozoal activity of pinocembrin and naringenin (IC50 of 29.51 µg/mL and 28.85 µg/mL, respectively) was higher compared with sakuranetin (44.47 µg/mL) and with cirsimaritin (150.00 µg/mL), revealing that a 5,7-dihydroxylated A ring is essential for antiprotozoal activity. These research funds may validate the use of this plant in the traditional Mexican medicine for the treatment of some digestive disorders and can help to integrate the use of extracts of L. graveolens in the conventional and complementary medicine for the treatment of parasitic diseases.

Keywords: amoebiasis, antiprotozoal agents, bioguided isolation, infectious diseases

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633 2D and 3D Breast Cancer Cells Behave Differently to the Applied Free Palbociclib or the Palbociclib-Loaded Nanoparticles

Authors: Maryam Parsian, Pelin Mutlu, Ufuk Gunduz

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Two-dimensional cell culture affords simplicity and low cost, but it has serious limitations; lacking cell-cell and cell-matrix interactions that are present in tissues. Cancer cells grown in 3D culture systems have distinct phenotypes of adhesion, growth, migration, invasion as well as profiles of gene and protein expression. These interactions cause the 3D-cultured cells to acquire morphological and cellular characteristics relevant to in vivo tumors. Palbociclib is a chemotherapeutic agent for the treatment of ER-positive and HER-negative metastatic breast cancer. Poly-amidoamine (PAMAM) dendrimer is a well-defined, special three-dimensional structure and has a multivalent surface and internal cavities that can play an essential role in drug delivery systems. In this study, palbociclib is loaded onto the magnetic PAMAM dendrimer. Hanging droplet method was used in order to form 3D spheroids. The possible toxic effects of both free drug and drug loaded nanoparticles were evaluated in 2D and 3D MCF-7, MD-MB-231 and SKBR-3 breast cancer cell culture models by performing MTT cell viability and Alamar Blue assays. MTT analysis was performed with six different doses from 1000 µg/ml to 25 µg/ml. Drug unloaded PAMAM dendrimer did not demonstrate significant toxicity on all breast cancer cell lines. The results showed that 3D spheroids are clearly less sensitive than 2D cell cultures to free palbociclib. Also, palbociclib loaded PAMAM dendrimers showed more toxic effect than free palbociclib in all cell lines at 2D and 3D cultures. The results suggest that the traditional cell culture method (2D) is insufficient for mimicking the actual tumor tissue. The response of the cancer cells to anticancer drugs is different in the 2D and 3D culture conditions. This study showed that breast cancer cells are more resistant to free palbociclib in 3D cultures than in 2D cultures. However, nanoparticle loaded drugs can be more cytotoxic when compared to free drug.

Keywords: 2D and 3D cell culture, breast cancer, palbociclibe, PAMAM magnetic nanoparticles

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632 Single and Combined Effects of Diclofenac and Ibuprofen on Daphnia Magna and Some Phytoplankton Species

Authors: Ramatu I. Sha’aba, Mathias A. Chia, Abdullahi B. Alhassan, Yisa A. Gana, Ibrahim M. Gadzama

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Globally, Diclofenac (DLC) and Ibuprofen (IBU) are the most prescribed drugs due to their antipyretic and analgesic properties. They are, however, highly toxic at elevated doses, with the involvement of an already described oxidative stress pathway. As a result, there is rising concern about the ecological fate of analgesics on non-target organisms such as Daphnia magna and Phytoplankton species. Phytoplankton is a crucial component of the aquatic ecosystem that serves as the primary producer at the base of the food chain. However, the increasing presence and levels of micropollutants such as these analgesics can disrupt their community structure, dynamics, and ecosystem functions. This study presents a comprehensive series of the physiology, antioxidant response, immobilization, and risk assessment of Diclofenac and Ibuprofen’s effects on Daphnia magna and the Phytoplankton community using a laboratory approach. The effect of DLC and IBU at 27.16 µg/L and 20.89 µg/L, respectively, for a single exposure and 22.39 µg/L for combined exposure of DLC and IBU for the experimental setup. The antioxidant response increased with increasing levels of stress. The highest stressor to the organism was 1000 µg/L of DLC and 10,000 µg/L of IBU. Peroxidase and glutathione -S-transferase activity was higher for Diclofenac + Ibuprofen. The study showed 60% and 70% immobilization of the organism at 1000 g L-1 of DLC and IBU. The two drugs and their combinations adversely impacted Phytoplankton biomass with increased exposure time. However, combining the drugs resulted in more significant adverse effects on physiological and pigment content parameters. The risk assessment calculation for the risk quotient and toxic unit of the analgesic reveals from this study was RQ Diclofenac = 8.41, TU Diclofenac = 3.68, and RQ Ibuprofen = 718.05 and TU Ibuprofen = 487.70. Hence, these findings demonstrate that the current exposure concentrations of Diclofenac and Ibuprofen can immobilize D. magna. This study shows the dangers of multiple drugs in the aquatic environment because their combinations could have additive effects on the structure and functions of Phytoplankton and are capable of immobilizing D. magna.

Keywords: algae, analgesic drug, daphnia magna, toxicity

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631 Efficacy of Topical Ectoin Therapy for Acute Radiodermatitis Associated with Breast Cancer Radiotherapy: A Randomized Controlled Study

Authors: Nagwa E. Abd Elazim, Maha S. El-naggar, Rania H. Mohamed, Sara M. Awad

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Background: Radiodermatitis is a common side effect of radiation therapy for breast cancer. However, there is no current consensus about effective standard therapy for the prevention and management of radiation dermatitis. Topical ectoine has demonstrated efficacy in the treatment of atopic dermatitis owing to its anti-inflammatory activity. Objective: To evaluate the efficacy of topical ectoine in comparison to traditional topical dexpanthenol treatment in the management of acute radiodermatitis in breast cancer patients undergoing adjuvant radiotherapy. Methods: Fifty patients were randomized to use either dexpanthenol 0.5% cream (25 patients), or ectoin 7% cream (25 patients), applied twice daily to the irradiated area during the radiation period and continued for 2 weeks after cessation of radiotherapy. Assessment of radiation skin toxicity using Common Terminology Criteria of Adverse Events (CTCAE) v4.0, radiation-associated symptoms, and adverse events were undertaken weekly during radiotherapy and 2 weeks after the end of radiotherapy. Results: Topical ectoine showed some clinical benefit over dexpanthenol, as shown by delayed time to onset (at week 3 versus week 2, respectively) and larger number of patients who reached grade 0 at the end of treatment (64% vs. 48%, respectively). The clinical symptoms of pain (p = 0.003) and itching (p = 0.001) attributable to radiation were less pronounced with ectoine than with dexpanthenol. Burning and hyperpigmentation were the most common side effects with ectoine. However, no significant difference between dexpanthenol and ectoine treatments was found in any of the side effects (p = 0.1). Conclusion: Ectoin was overall more effective in improving radiation dermatitis than topical dexpanthenol in breast cancer patients. Ectoin could be proposed as a preventive or curative treatment for patients undergoing postoperative irradiation for breast cancer. Further clinical studies with a larger number of patients are recommended for the confirmation of these preliminary results.

Keywords: breast cancer, dexapanthenol, ectoin, radiation dermatitis

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630 Optimization, Characterization and Stability of Trachyspermum copticum Essential Oil Loaded in Niosome Nanocarriers

Authors: Mohadese Hashemi, Elham Akhoundi Kharanaghi, Fatemeh Haghiralsadat, Mojgan Yazdani, Omid Javani, Mahboobe Sharafodini, Davood Rajabi

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Niosomes are non-ionic surfactant vesicles in aqueous media resulting in closed bilayer structures that can be used as carriers of hydrophilic and hydrophobic compounds. The use of niosomes for encapsulation of essential oils (EOs) is an attractive new approach to overcome their physicochemical stability concerns include sensibility to oxygen, light, temperature, and volatility, and their reduced bioavailability which is due to low solubility in water. EOs are unstable and fragile volatile compounds which have strong interest in pharmaceutical due to their medicinal properties such as antiviral, anti-inflammatory, antifungal, and antioxidant activities without side effects. Trachyspermum copticum (ajwain) is an annual aromatic plant with important medicinal properties that grows widely around Mediterranean region and south-west Asian countries. The major components of the ajwain oil were reported as thymol, γ-terpinene, p-cymene, and carvacrol which provide antimicrobial and antioxidant activity. The aim of this work was to formulate ajwain essential oil-loaded niosomes to improve water solubility of natural product and evaluate its physico-chemical features and stability. Ajwain oil was obtained through steam distillation using a clevenger-type apparatus and GC/MS was applied to identify the main components of the essential oil. Niosomes were prepared by using thin film hydration method and nanoparticles were characterized for particle size, dispersity index, zeta potential, encapsulation efficiency, in vitro release, and morphology.

Keywords: trachyspermum copticum, ajwain, niosome, essential oil, encapsulation

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629 Exploring the Design of Prospective Human Immunodeficiency Virus Type 1 Reverse Transcriptase Inhibitors through a Comprehensive Approach of Quantitative Structure Activity Relationship Study, Molecular Docking, and Molecular Dynamics Simulations

Authors: Mouna Baassi, Mohamed Moussaoui, Sanchaita Rajkhowa, Hatim Soufi, Said Belaaouad

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The objective of this paper is to address the challenging task of targeting Human Immunodeficiency Virus type 1 Reverse Transcriptase (HIV-1 RT) in the treatment of AIDS. Reverse Transcriptase inhibitors (RTIs) have limitations due to the development of Reverse Transcriptase mutations that lead to treatment resistance. In this study, a combination of statistical analysis and bioinformatics tools was adopted to develop a mathematical model that relates the structure of compounds to their inhibitory activities against HIV-1 Reverse Transcriptase. Our approach was based on a series of compounds recognized for their HIV-1 RT enzymatic inhibitory activities. These compounds were designed via software, with their descriptors computed using multiple tools. The most statistically promising model was chosen, and its domain of application was ascertained. Furthermore, compounds exhibiting comparable biological activity to existing drugs were identified as potential inhibitors of HIV-1 RT. The compounds underwent evaluation based on their chemical absorption, distribution, metabolism, excretion, toxicity properties, and adherence to Lipinski's rule. Molecular docking techniques were employed to examine the interaction between the Reverse Transcriptase (Wild Type and Mutant Type) and the ligands, including a known drug available in the market. Molecular dynamics simulations were also conducted to assess the stability of the RT-ligand complexes. Our results reveal some of the new compounds as promising candidates for effectively inhibiting HIV-1 Reverse Transcriptase, matching the potency of the established drug. This necessitates further experimental validation. This study, beyond its immediate results, provides a methodological foundation for future endeavors aiming to discover and design new inhibitors targeting HIV-1 Reverse Transcriptase.

Keywords: QSAR, ADMET properties, molecular docking, molecular dynamics simulation, reverse transcriptase inhibitors, HIV type 1

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628 Development of Site-Specific Colonic Drug Delivery System (Nanoparticles) of Chitosan Coated with pH Sensitive Polymer for the Management of Colonic Inflammation

Authors: Pooja Mongia Raj, Rakesh Raj, Alpana Ram

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Background: The use of multiparticulate drug delivery systems in preference to single unit dosage forms for colon targeting purposes dates back to 1985 when Hardy and co-workers showed that multiparticulate systems enabled the drug to reach the colon quickly and were retained in the ascending colon for a relatively long period of time. Methods: Site-specific colonic drug delivery system (nanoparticles) of 5-ASA were prepared and coated with pH sensitive polymer. Chitosan nanoparticles (CTNP) bearing 5-Amino salicylic acid (5-ASA) were prepared, by ionotropic gelation method. Nanoparticulate dosage form consisting of a hydrophobic core enteric coated with pH-dependent polymer Eudragit S-100 by solvent evaporation method, for the effective delivery of drug to the colon for treatment of ulcerative colitis. Results: The mean diameter of CTNP and ECTNP formulations were 159 and 661 nm, respectively. Also optimum value of polydispersity index was found to be 0.249 [count rate (kcps) was 251.2] and 0.170 [count rate (kcps) was 173.9] was obtained for both the formulations respectively. Conclusion: CTNP and Eudragit chitosan nanoparticles (ECTNP) was characterized for shape and surface morphology by scanning electron microscopy (SEM) appeared to be spherical in shape. The in vitro drug release was investigated using USP dissolution test apparatus in different simulated GIT fluids showed promising release. In vivo experiments are in further proceeding for fruitful results.

Keywords: colon targeting, nanoparticles, polymer, 5-amino salicylic acid, edragit

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627 Mechanisms and Regulation of the Bi-directional Motility of Mitotic Kinesin Nano-motors

Authors: Larisa Gheber

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Mitosis is an essential process by which duplicated genetic information is transmitted from mother to daughter cells. Incorrect chromosome segregation during mitosis can lead to genetic diseases, chromosome instability and cancer. This process is mediated by a dynamic microtubule-based intracellular structure, the mitotic spindle. One of the major factors that govern the mitotic spindle dynamics are the kinesin-5 biological nano motors that were believed to move unidirectionally on the microtubule filaments, using ATP hydrolysis, thus performing essential functions in mitotic spindle dynamics. Surprisingly, several reports from our and other laboratories have demonstrated that some kinesin-5 motors are bi-directional: they move in minus-end direction on the microtubules as single-molecules and can switch directionality under a number of conditions. These findings broke a twenty-five-years old dogma regarding kinesin directionality (1, 2). The mechanism of this bi-directional motility and its physiological significance remain unclear. To address this unresolved problem, we apply an interdisciplinary approach combining live cell imaging, biophysical single molecule, and structural experiments to examine the activity of these motors and their mutated variants in vivo and in vitro. Our data shows that factors such as protein phosphorylation (3, 4), motor clustering on the microtubules (5, 6) and structural elements (7, 8) regulate the bi-directional motility of kinesin motors. We also show, using Cryo-EM, that bi-directional kinesin motors obtain non-canonical microtubule binding, which is essential to their special motile properties and intracellular functions. We will discuss the implication of these findings to mechanism bi-directional motility and physiological roles in mitosis.

Keywords: mitosis, cancer, kinesin, microtubules, biochemistry, biophysics

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626 Biological Treatment of Corn Stover with Pleurotus ostreatus, Pleurotus eryngii and Lentinula edudes to Improve Digestibility

Authors: Aydan Atalar, Nurcan Cetinkaya

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Corn stover is leftover of the leaves, stalk, husks and tassels in the field after harvesting the grain combined. Corn stover is a low-quality roughage but has mostly been used as roughage source for feeding ruminant animals in developing countries including Turkey; however, it can also be used to make biofuels as in developed countries. The objectives of the present study were to improve the digestibility of corn stover by the treatment of white rod fungus mainly Pleurotus osteritus (PO), Pleurotus eryingii (PE) and Lantinula edudes (LE) at different incubation times and also to determine the most effective fungus and incubation time to prepare fermeted corn stover for ruminant nutrition. The choped corn stover was treated with PO, PE and LE and incubated for 10, 20, 30 and 40 days in incubator at 26 0C. After each incubation time dry matter(DM), organic matter(OM), crude protein (CP), neutral detergent fiber (NDF), acid detergent fiber (ADF), neutral detergent lignin (ADL), in-vitro true dry matter digestibility (IVTDMD) and organic matter digestibility (IVTOMD) were determined. The mean IVTDMD and IVTOMD levels were increased by PO, PE and LE treatments in increasing order of incubation times. The obtained IVTDM values were 59.45, 60.51, 60.82 and 60.18 %; 59.45, 70.55, 67.18 and 66.96 %; 59.45, 70.55, 67.18 and 66,96 %; 59.45, 74.90, 69.18 % ; 59.45, 76.50, 71.24 and 73.04 for control, PO, PE and LE treatments at 0, 10, 20, 30 and 40 days incubation times respectively. The obtained IVTOMD values were 56.45,60.26,60.82and 60.18 %; 56.45, 68.70, 67.18 and 66.96 %; 56.45, 71.26, 69.18 and 69.28 %; 56.45, 73.23, 71.24 and 73.04 % for control, PO, PE and LE treatments at 0, 10, 20, 30 and 40 days incubation times respectively. The most effective fungus was PO and the incubation time was 30 days. In conclusion, PO treatment of corn stover with 30 days incubation may be used to prepare fermented corn stover for ruminant nutrition.

Keywords: biological treatment, corn stover, digestibility, Lantinula edudes, Pleurotus eryingii, Pleurotus osteritus

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625 Pollution Associated with Combustion in Stove to Firewood (Eucalyptus) and Pellet (Radiate Pine): Effect of UVA Irradiation

Authors: Y. Vásquez, F. Reyes, P. Oyola, M. Rubio, J. Muñoz, E. Lissi

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In several cities in Chile, there is significant urban pollution, particularly in Santiago and in cities in the south where biomass is used as fuel in heating and cooking in a large proportion of homes. This has generated interest in knowing what factors can be modulated to control the level of pollution. In this project was conditioned and set up a photochemical chamber (14m3) equipped with gas monitors e.g. CO, NOX, O3, others and PM monitors e.g. dustrack, DMPS, Harvard impactors, etc. This volume could be exposed to UVA lamps, producing a spectrum similar to that generated by the sun. In this chamber, PM and gas emissions associated with biomass burning were studied in the presence and absence of radiation. From the comparative analysis of wood stove (eucalyptus globulus) and pellet (radiata pine), it can be concluded that, in the first approximation, 9-nitroanthracene, 4-nitropyrene, levoglucosan, water soluble potassium and CO present characteristics of the tracers. However, some of them show properties that interfere with this possibility. For example, levoglucosan is decomposed by radiation. The 9-nitroanthracene, 4-nitropyrene are emitted and formed under radiation. The 9-nitroanthracene has a vapor pressure that involves a partition involving the gas phase and particulate matter. From this analysis, it can be concluded that K+ is compound that meets the properties known to be tracer. The PM2.5 emission measured in the automatic pellet stove that was used in this thesis project was two orders of magnitude smaller than that registered by the manual wood stove. This has led to encouraging the use of pellet stoves in indoor heating, particularly in south-central Chile. However, it should be considered, while the use of pellet is not without problems, due to pellet stove generate high concentrations of Nitro-HAP's (secondary organic contaminants). In particular, 4-nitropyrene, compound of high toxicity, also primary and secondary particulate matter, associated with pellet burning produce a decrease in the size distribution of the PM, which leads to a depth penetration of the particles and their toxic components in the respiratory system.

Keywords: biomass burning, photochemical chamber, particulate matter, tracers

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624 Anticancer Effect of Resveratrol-Loaded Gelatin Nanoparticles in NCI-H460 Non-Small Cell Lung Carcinoma Cell Lines

Authors: N. Rajendra Prasad

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Resveratrol (RSV), a grape phytochemical, has drawn greater attention because of its beneficial ef-fects against cancer. However, RSV has some draw-backs such as unstabilization, poor water solubility and short biological half time, which limit the utili-zation of RSV in medicine, food and pharmaceutical industries. In this study, we have encapsulated RSV in gelatin nanoparticles (GNPs) and studied its anti-cancer efficacy in NCI-H460 lung cancer cells. SEM and DLS studies have revealed that the prepared RSV-GNPs possess spherical shape with a mean diameter of 294 nm. The successful encapsulation of RSV in GNPs has been achieved by the cross-linker glutaraldehyde probably through Schiff base reaction and hydrogen bond interaction. Spectrophotometric analysis revealed that the max-imum of 93.6% of RSV has been entrapped in GNPs. In vitro drug release kinetics indicated that there was an initial burst release followed by a slow and sustained release of RSV from GNPs. The prepared RSV-GNPs exhibited very rapid and more efficient cellular uptake than free RSV. Further, RSV-GNPs treatment showed greater antiproliferative efficacy than free RSV treatment in NCI-H460 cells. It has been found that greater ROS generation, DNA damage and apoptotic incidence in RSV-GNPs treated cells than free RSV treatment. Erythrocyte aggregation assay showed that the prepared RSV-GNPs formulation elicit no toxic response. HPLC analysis revealed that RSV-GNPs was more bioavailable and had a longer half-life than free RSV. Hence, GNPs carrier system might be a promising mode for controlled delivery and for improved therapeutic index of poorly water soluble RSV.

Keywords: resveratrol, coacervation, anticancer gelatin nanoparticles, lung cancer, controlled release

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623 The Protective Role of Decoy Receptor 3 Analogue on Rat Steatotic Liver against Ischemia-Reperfusion Injury by Blocking M1/Th1 Polarization and Multiple Upstream Pathogenic Cascades

Authors: Tzu-Hao Li, Shie-Liang Hsieh, Han-Chieh Lin, Ying-Ying Yang

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TNF superfamily-stimulated pathogenic cascades and macrophage (M1)/kupffer cells (KC) polarization are important in the pathogenesis of ischemia-reperfusion (IR) liver injury in animals with hepatic steatosis (HS). Decoy receptor 3 (DcR3) is a common upstream inhibitor of the above-mentioned pathogenic cascades. The study evaluated whether modulation of these DcR3-related cascades was able to protect steatotic liver from IR injury. Serum and hepatic DcR3 levels were lower in patients and animals with HS. Accordingly, the effects of pharmacologic and genetic DcR3 replacement on the IR-related pathogenic changes were measured. Significantly, DcR3 replacement protected IR-Zucker(HS) rats and IR-DcR3-Tg(HS) mice from IR liver injury. The beneficial effects of DcR3 replacement were accompanied by decreased serum/hepatic TNF, soluble TNF-like cytokine 1A (TL1A), Fas ligand (Fas-L) and LIGHT, T-helper-cell-1 cytokine (INF) levels, neutrophil infiltration, M1 polarization, neutrophil-macrophage/KC-T-cell interaction, hepatocyte apoptosis and improved hepatic microcirculatory failure among animals with IR-injured steatotic livers. Additionally, TL1A, Fas-L, LIGHT and TLR4/NFB signals were found to mediate the DcR3-related protective effects of steatotic livers from IR injury. Using multimodal in vivo and in vitro approaches, we found that DcR3 was a potential agent to protect steatotic livers from IR injury by simultaneous blocking the multiple IR injury-related pathogenic changes.

Keywords: Decoy 3 receptor, ischemia-reperfusion injury, M1 polarization, TNF superfamily

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622 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

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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

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621 System for Mechanical Stimulation of the Mesenchymal Stem Cells Supporting Differentiation into Osteogenic Cells

Authors: Jana Stepanovska, Roman Matejka, Jozef Rosina, Marta Vandrovcova, Lucie Bacakova

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The aim of this study was to develop a system for mechanical and also electrical stimulation controlling in vitro osteogenesis under conditions more similar to the in vivo bone microenvironment than traditional static cultivation, which would achieve good adhesion, growth and other specific behaviors of osteogenic cells in cultures. An engineered culture system for mechanical stimulation of the mesenchymal stem cells on the charged surface was designed. The bioreactor allows efficient mechanical loading inducing an electrical response and perfusion of the culture chamber with seeded cells. The mesenchymal stem cells were seeded to specific charged materials, like polarized hydroxyapatite (Hap) or other materials with piezoelectric and ferroelectric features, to create electrical potentials for stimulating of the cells. The material of the matrix was TiNb alloy designed for these purposes, and it was covered by BaTiO3 film, like a kind of piezoelectric material. The process of mechanical stimulation inducing electrical response is controlled by measuring electrical potential in the chamber. It was performed a series of experiments, where the cells were seeded, perfused and stimulated up to 48 hours under different conditions, especially pressure and perfusion. The analysis of the proteins expression was done, which demonstrated the effective mechanical and electrical stimulation. The experiments demonstrated effective stimulation of the cells in comparison with the static culture. This work was supported by the Ministry of Health, grant No. 15-29153A and the Grant Agency of the Czech Republic grant No. GA15-01558S.

Keywords: charged surface, dynamic cultivation, electrical stimulation, ferroelectric layers, mechanical stimulation, piezoelectric layers

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620 Carbon-Encapsulated Iron Nanoparticles for Hydrogen Sulfide Removal

Authors: Meriem Abid, Erika Oliveria-Jardim, Andres Fullana, Joaquin Silvestre-Albero

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The rapid industrial development associated with the increase of volatile organic compounds (VOCs) has seriously impacted the environment. Among VOCs, hydrogen sulfide (H₂S) is known as a highly toxic, malodorous, flammable, and corrosive gas, which is emitted from diverse chemical processes, including industrial waste-gas streams, natural gas processing, and biogas purification. The high toxicity, corrosively, and very characteristic odor threshold of H2S call for urgent development of efficient desulfurization processes from the viewpoint of environmental protection and resource regeneration. In order to reduce H₂S emissions, effective technologies for have been performed. The general method of H₂S removal included amine aqueous solution, adsorption process, biological methods, and fixed-bed solid catalytic oxidation processes. Ecologically and economically, low-temperature direct oxidation of H₂S to elemental sulfur using catalytic oxidation is the preferred approach for removing H₂S-containing gas streams. A large number of catalysts made from carbon, metal oxides, clay, and others, have been studied extensively for this application. In this sense, activated carbon (AC) is an attractive catalyst for H₂S removal because it features a high specific surface area, diverse functional groups, low cost, durability, and high efficiency. It is interesting to stand out that AC is modified using metal oxides to promote the efficiency of H₂S removal and to enhance the catalytic performance. Based on these premises, the main goal of the present study is the evaluation of the H₂S adsorption performance in carbon-encapsulated iron nanoparticles obtained from an olive mill, thermally treated at 600, 800 and 1000 ºC temperatures under anaerobic conditions. These results anticipate that carbon-encapsulated iron nanoparticles exhibit a promising performance for the H₂S removal up to 360 mg/g.

Keywords: H₂S removal, catalytic oxidation, carbon encapsulated iron, olive mill wastewater

Procedia PDF Downloads 88
619 Wound Healing and Antioxidant Properties of 80% Methanol Leaf Extract of Verbascum sinaiticum (Scrophulariaceae), an Ethiopian Medicinal Plant

Authors: Solomon Assefa Huluka

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Wounds account for severe morbidity, socioeconomic distress, and mortality around the globe.For several years, various herbal products are used to expediteand augment the innate wound healing process. In Ethiopian folkloricmedicine, Verbascum sinaiticum L. (V. sinaiticum) is commonlyapplied as a wound-healing agent. The present study investigated the potential wound healing and antioxidant properties of hydroalcoholic leaf extract of V. sinaiticum. The 80% methanol extract, formulated as 5% (w/w) and 10% (w/w) ointments, was evaluated in excision and incision wound models using nitrofurazone and simple ointment as positive and negative controls, respectively. Parameters such as wound contraction, period of epithelialization, and tensile strength were determined. Moreover, its in vitro antioxidant property was evaluated using a DPPH assay. In the excision model, both doses (5% and 10% w/w) of the extract showed a significant (p<0.001) wound healing efficacy compared to the negative control, as evidenced by enhanced wound contraction rate and shorter epithelialization time records. In the incision model, the lower dose (5% w/w) ointment formulation of the extract exhibited the maximum increment in tensile strength (85.6%) that was significant (p<0.001)compared to negative and untreated controls. Animals treated with 5% w/wointment, furthermore, showed a significantly (p < 0.05) higher percentage of tensile strength than nitrofurazone treated ones. Moreover, the hydroalcoholic extract of the plant showed a noticeable free radical scavenging property. The result of the present study upholds the folkloric use of V. sinaiticum in the treatment of wounds.

Keywords: wound healing, antioxidant, excision wound model, incision wound model, verbascum sinaiticum

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618 CP-96345 Rregulates Hydrogen Sulphide Induced TLR4 Signaling Pathway Adhesion Molecules in Caerulein Treated Pancreatic Acinar Cells

Authors: Ramasamy Tamizhselvi, Leema George, Madhav Bhatia

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We have earlier shown that mouse pancreatic acinar cells produce hydrogen sulfide (H2S) and play a role in the pathogenesis of acute pancreatitis. This study is to determine the effect of H2S on TLR4 mediated innate immune signaling in acute pancreatitis via substance P (SP). Male Swiss mice were treated with hourly intraperitoneal injection of caerulein (50μg/kg) for 10 hour. DL-propargylglycine (PAG) (100 mg/kg i.p.), an inhibitor of H2S formation was administered 1h after the induction of acute pancreatitis. Pancreatic acinar cells from male Swiss mice were incubated with or without caerulein (10–7 M for 60 min) and CP-96345 (NK1R inhibitor). To better understand the effect of H2S in inflammation, acinar cells were stimulated with caerulein after addition of H2S donor, NaHS. In addition, caerulein treated pancreatic acinar cells were pretreated with PAG (30 µM), for 1h. H2S inhibitor, PAG, eliminated TLR4, IRAK4, TRAF6 and NF-kB levels in an in vitro and in vivo model of caerulein-induced acute pancreatitis. PPTA gene deletion reduced TLR4, MyD88, IRAK4, TRAF6, adhesion molecules and NF-kB in caerulein treated pancreatic acinar cells whereas administration of NaHS resulted in further rise in TLR4 and NF-kB levels in caerulein treated pancreatic acinar cells. In addition, acini isolated from mice and treated with PPTA gene receptor NK1R antagonist CP96345 did not exhibit further increase in TLR4, IRAK4, TRAF6, adhesion molecules and NF-kB levels after NaHS pretreatment. The present findings show for the first time that in acute pancreatitis, H2S up-regulates TLR4 pathway and NF-kB via substance P.

Keywords: preprotachykinin-A gene, H2S, TLR4, acute pancreatitis

Procedia PDF Downloads 278
617 The Activity of Polish Propolis and Cannabidiol Oil Extracts on Glioblastoma Cell Lines

Authors: Sylwia K. Naliwajko, Renata Markiewicz-Zukowska, Justyna Moskwa, Krystyna Gromkowska-Kepka, Konrad Mielcarek, Patryk Nowakowski, Katarzyna Socha, Anna Puscion-Jakubik, Maria H. Borawska

Abstract:

Glioblastoma (grade IV WHO) is a rapidly progressive brain tumor with very high morbidity and mortality. The vast malignant gliomas are not curable despite the therapy (surgical, radiotherapy, chemotherapy) and patients seek alternative or complementary treatments. Patients often use cannabidiol (CBD) oil as an alternative therapy of glioblastoma. CBD is one of the cannabinoids, an active component of Cannabis sativa. THC (Δ9-tetrahydrocannabinol) can be addictive, and in many countries CBD oil without THC ( < 0,2%) is available. Propolis produced by bees from the resin collected from trees has antiglioma properties in vitro and can be used as a supplement in complementary therapy of gliomas. The aim of this study was to examine the influence of extract from CBD oil in combination with propolis extract on two glioblastoma cell lines. The MTT (Thiazolyl Blue Tetrazolium Bromide) test was used to determine the influence of CBD oil extract and polish propolis extract (PPE) on the viability of glioblastoma cell lines – U87MG and LN18. The cells were incubated (24, 48 and 72 h) with CBD oil extract and PPE. CBD extract was used in concentration 1, 1.5 and 3 µM and PPE in 30 µg/mL. The data were presented compared to the control. The statistical analysis was performed using Statistica v. 13.0 software. CBD oil extract in concentrations 1, 1.5 and 3 µM did not inhibit the viability of U87MG and LN18 cells (viability more than 90% cells compared to the control). There was no dose-response viability, and IC50 value was not recognized. PPE in the concentration of 30 µg/mL time-dependently inhibited the viability of U87MG and LN18 cell line (after 48 h the viability as a percent of the control was 59,7±6% and 57,8±7%, respectively). In a combination of CBD with PPE, the viability of the treated cells was similar to PPE used alone (58,2±7% and 56,5±9%, respectively). CBD oil extract did not show anti-glioma activity and in combination with PPE did not change the activity of PPE.

Keywords: anticancer, cannabidiol, cell line, glioblastoma

Procedia PDF Downloads 246
616 Reduced Glycaemic Impact by Kiwifruit-Based Carbohydrate Exchanges Depends on Both Available Carbohydrate and Non-Digestible Fruit Residue

Authors: S. Mishra, J. Monro, H. Edwards, J. Podd

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When a fruit such as kiwifruit is consumed its tissues are released from the physical /anatomical constraints existing in the fruit. During digestion they may expand several-fold to achieve a hydrated solids volume far greater than the original fruit, and occupy the available space in the gut, where they surround and interact with other food components. Within the cell wall dispersion, in vitro digestion of co-consumed carbohydrate, diffusion of digestion products, and mixing responsible for mass transfer of nutrients to the gut wall for absorption, were all retarded. All of the foregoing processes may be involved in the glycaemic response to carbohydrate foods consumed with kiwifruit, such as breakfast cereal. To examine their combined role in reducing the glycaemic response to wheat cereal consumed with kiwifruit we formulated diets containing equal amounts of breakfast cereal, with the addition of either kiwifruit, or sugars of the same composition and quantity as in kiwifruit. Therefore, the only difference between the diets was the presence of non-digestible fruit residues. The diet containing the entire disperse kiwifruit significantly reduced the glycaemic response amplitude and the area under the 0-120 min incremental blood glucose response curve (IAUC), compared with the equicarbohydrate diet containing the added kiwifruit sugars. It also slightly but significantly increased the 120-180 min IAUC by preventing a postprandial overcompensation, indicating improved homeostatic blood glucose control. In a subsequent study in which we used kiwifruit in a carbohydrate exchange format, in which the kiwifruit carbohydrate partially replaced breakfast cereal in equal carbohydrate meals, the blood glucose was further reduced without a loss of satiety, and with a reduction in insulin demand. The results show that kiwifruit may be a valuable component in low glycaemic impact diets.

Keywords: carbohydrate, digestion, glycaemic response, kiwifruit

Procedia PDF Downloads 496
615 Covalent Binding of Cysteine to a Sol-Gel Material for Cadmium Biosorption from Aqueous Solutions

Authors: Claudiu Marcu, Cristina Paul, Adelina Andelescu, Corneliu Mircea Davidescu, Francisc Péter

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Heavy metal pollution has become a more serious environmental problem in the last several decades as a result of its toxicity and insusceptibility to the environment. Methods for removing metal ions from aqueous solution mainly consist of physical, chemical and biochemical procedures. Biosorption is defined as the removal of metal or metalloid species, compounds and particulates from solution by a biological material. Biosorption represents a very attractive method for the removal of toxic metal ions from aqueous effluents because it uses the ability of various biomass to bind the metal ions without the risk of releasing other toxic chemical compounds into the environment. The problem with using biomass or living cells as biosorbents is that their regeneration/reuse is often either impossible or very laborious. One of the most common chelating group found in biosorbents is the thiol group in cysteine. Therefore, we immobilized cysteine using covalent binding using glutaraldehyde as a linker on a synthetic sol-gel support obtained using 3-amino-propyl-trimetoxysilane and trimetoxysilane as precursors. The obtained adsorbents were used for removal of cadmium from aqueous solutions and the removal capacity was investigated in relation to the composition of the sol-gel hybrid composite, the loading of the biomolecule and the physical parameters of the biosorption process. In the same conditions, the bare sol-gel support without cysteine had no Cd removal effect, while the adsorbent with cysteine had an adsorption capacity up to 25.8 mg Cd/g adsorbent at pH 2.0 and 119 mg Cd/g adsorbent at pH 6.6, depending on cadmium concentration and adsorption conditions. We used atomic adsorption spectrometry to assess the cadmium concentration in the samples after the biosorbtion process. The parameters for the Freundlich and Langmuir adsorption isotherms where calculated from plotting the results of the adsorption experiments. The results for cysteine immobilization show a good loading capacity of the sol-gel support which indicates it could be used to immobilize metal binding proteins and by doing so boosting the heavy metal adsorption capacity of the biosorbent.

Keywords: biosorbtion, cadmium, cysteine covalent binding, sol-gel

Procedia PDF Downloads 294
614 Screening for Enterotoxigenic Staphylococcus spp. Strains Isolated From Raw Milk and Dairy Products in R. N. Macedonia

Authors: Marija Ratkova Manovska, Mirko Prodanov, Dean Jankuloski, Katerina Blagoevska

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Staphylococci, which are widely found in the environment, animals, humans, and food products, include Staphylococcus aureus (S. aureus), the most significant pathogenic species in this genus. The virulence and toxicity of S. aureus are primarily attributed to the presence of specific genes responsible for producing toxins, biofilms, invasive components, and antibiotic resistance. Staphylococcal food poisoning, caused by the production of staphylococcal enterotoxins (SEs) by these strains in food, is a common occurrence. Globally, S. aureus food intoxications are typically ranked as the third or fourth most prevalent foodborne intoxications. For this study, a total of 333 milk samples and 1160 dairy product samples were analyzed between 2016 and 2020. The strains were isolated and confirmed using the ISO 6888-1:1999 "Horizontal method for enumeration of coagulase-positive staphylococci." Molecular analysis of the isolates, conducted using conventional PCR, involved detecting the 23s gene of S. aureus, the nuc gene, the mecA gene, and 11 genes responsible for producing enterotoxins (sea, seb, sec, sed, see, seg, seh, sei, ser, sej, and sep). The 23s gene was found in 93 (75.6%) out of 123 isolates of Staphylococcus spp. obtained from milk. Among the 76 isolates from dairy products, either S. aureus or the 23s gene was detected in 49 (64.5%) of them. The mecA gene was identified in three isolates from raw milk and five isolates from cheese samples. The nuc gene was present in 98.9% of S. aureus strains from milk and 97.9% from dairy products. Other Staphylococcus strains carried the nuc gene in 26.7% of milk strains and 14.8% of dairy product strains. Genes associated with SEs production were detected in 85 (69.1%) strains from milk and 38 (50%) strains from dairy products. In this study, 10 out of the 11 SEs genes were found, with no isolates carrying the see gene. The most prevalent genes detected were seg and sei, with some isolates containing up to five different SEs genes. These findings indicate the presence of enterotoxigenic staphylococci strains in the tested samples, emphasizing the importance of implementing proper sanitation and hygienic practices, utilizing safe raw materials, and ensuring adequate handling of finished products. Continued monitoring for the presence of SEs is necessary to ensure food safety and prevent intoxication.

Keywords: dairy products, milk, Staphylococci, enterotoxins, SE genes

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613 Smart Coating for Enhanced Corneal Healing via Delivering Progranulin

Authors: Dan Yan, Yunuo Zhang, Yuhan Huang, Weijie Ouyang

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The cornea serves as a vital protective barrier for the eye; however, it is prone to injury and damage that can disrupt corneal epithelium and nerves, triggering inflammation. Therefore, understanding the biological effects and molecular mechanisms involved in corneal wound healing and identifying drugs targeting these pathways is crucial for researchers in this field. This study aimed to investigate the therapeutic potential of progranulin (PGRN) in treating corneal injuries. Our findings demonstrated that PGRN significantly enhanced corneal wound repair by accelerating corneal re-epithelialization and re-innervation. In vitro experiments with cultured epithelial cells and trigeminal ganglion cells further revealed that PGRN stimulated corneal epithelial cell proliferation and promoted axon growth in trigeminal ganglion cells. Through RNA-sequencing (RNA-seq) analysis and other experimental techniques, we discovered that PGRN exerted its healing effects by modulating the Wnt signaling pathway, which played a critical role in repairing epithelial cells and promoting axon regeneration in trigeminal neurons. Importantly, our study highlighted the anti-inflammatory properties of PGRN by inhibiting the NF-κB signaling pathway, leading to decreased infiltration of macrophages. In conclusion, our findings underscored the potential of PGRN in facilitating corneal wound healing by promoting corneal epithelial cell proliferation, trigeminal ganglion cell axon regeneration, and suppressing ocular inflammation. These results suggest that PGRN could potentially expedite the healing process and improve visual outcomes in patients with corneal injuries.

Keywords: cornea, wound healing, progranulin, corneal epithelial cells, trigeminal ganglion cells

Procedia PDF Downloads 58
612 Transcriptomic Response of Calmodulin Encoding Gene (CaM) in Pesticide Utilizing Talaromyces Fungal Strains

Authors: M. D. Asemoloye, S. G. Jonathan, A. Rafiq, O. J. Olawuyi, D. O. Adejoye

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Calmodulin is one of the intracellular calcium proteins that regulates large spectrum of enzymes and cellular functions including metabolism of cyclic nucleotides and glycogen. The potentials of calmodulin gene in fungi necessitates their genetic response and their strong cassette of enzyme secretions for pesticide degradation. Therefore, this study was carried out to investigate the ‘Transcriptomic’ response of calmodulin encoding genes in Talaromyces fungi in response to 2, 2-dichlorovinyl dimethyl phosphate (DDVP or Dichlorvos) an organophosphate pesticide and γ-Hexachlorocyclohexane (Lindane) an organochlorine pesticide. Fungi strains isolated from rhizosphere from grasses rhizosphere in pesticide polluted sites were subjected to percentage incidence test. Two most frequent fungi were further characterized using ITS gene amplification (ITS1 and ITS4 combinations), they were thereafter subjected to In-vitro DDVP and lindane tolerance tests at different concentrations. They were also screened for presence and expression of calmodulin gene (caM) using RT-PCR technique. The two Talaromyces strains had the highest incidence of 50-72% in pesticide polluted site, they were both identified as Talaromyces astroroseus asemoG and Talaromyces purpurogenum asemoN submitted in NCBI gene-bank with accession numbers KY488464 and KY488468 respectively. T. astroroseus KY488464 tolerated DDVP (1.23±0.023 cm) and lindane (1.11±0.018 cm) at 25 % concentration while T. purpurogenum KY488468 tolerated DDVP (1.33±0.061 cm) and lindane (1.54±0.077 cm) at this concentration. Calmodulin gene was detected in both strains, but RT-PCR expression of caM gene revealed at 900-1000 bp showed an under-expression of caM in T. astrorosues KY488464 but overexpressed in T. purpurogenum KY488464. Thus, the calmodulin gene response of these fungal strains to both pesticides could be considered in monitoring the potentials of fungal strains to pesticide tolerance and bioremediation of pesticide in polluted soil.

Keywords: Calmodulin gene, pesticide, RT-PCR, talaromyces, tolerance

Procedia PDF Downloads 225
611 Chemical Modifications of Carotol and Their Antioxidant Activity

Authors: Dalvir Kataria, Khushminder Kaur Chahal, Amit Kumar

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The carrot seed essential oil was obtained by hydrodistillation. Hexane, dichloromethane, and methanol solvents were used for extraction of carrot seed by Soxhlet extraction methods. The major and minor compounds identified in carrot seed essential oil were carotol (52.73), daucol (5.10), daucene (5.68), (E)-β-farnesene (5.40), β-cubebene (3.19), longifolenaldehyde (3.23), β-elimene (3.23), (E)-caryophyllene (1.22), β-bisabolene (2.95) etc. The chemical composition of hexane, dichloromethane, and methanol extracts was different. Carotol was the common compound present. Major compounds isolated were from the carrot seed essential oil by column chromatography. Chemical transformations of carotol (2) with mercuric acetate/sodium borohydride, dry hydrochloric acid gas, acetonitrile/sulfuric acid, selenium dioxide/t-butyl hydrogen peroxide, N-bromosuccinimide, hydrogen iodide, and phenol were carried out. The derivatives of carotol were designed to explore the significance of some structural modifications in relation to antioxidant activities. The structures of major compounds and derivatives were confirmed on the basis of FT-IR, 1HNMR and 13CNMR spectroscopy. Antioxidant activity of carrot seed essential oil, various extracts and isolated compounds were tested by in vitro models involving 2, 2-diphenyl-1-picrylhydrazyl (DPPH•), hydroxyl (OH•), nitric oxide (NO•), superoxide radical scavenging methods and ferric reducing antioxidant power assay (FRAP). Chemical transformations of major isolated compound carotol were carried out, and antioxidant activity of all compounds was undertaken. The major sesquiterpenoidcarotol isolated from carrot seed essential oil showed the highest antioxidant activity in all the methods. The methanol extract showed higher antioxidant potential as compared to carrot seed essential oil, hexane, and dichloromethane extracts.

Keywords: antioxidant, carotol, carrot, DPPH

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610 Potential Application of Selected Halotolerant PSB Isolated from Rhizospheric Soil of Chenopodium quinoa in Plant Growth Promotion

Authors: Ismail Mahdi, Nidal Fahsi, Mohamed Hafidi, Abdelmounaim Allaoui, Latefa Biskri

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To meet the worldwide demand for food, smart management of arable lands is needed. This could be achieved through sustainable approaches such as the use of plant growth-promoting microorganisms including bacteria. Phosphate (P) solubilization is one of the major mechanisms of plant growth promotion by associated bacteria. In the present study, we isolated and screened 14 strains from the rhizosphere of Chenopodium quinoa wild grown in the experimental farm of UM6P and assessed their plant growth promoting properties. Next, they were identified by using 16S rRNA and Cpn60 genes sequencing as Bacillus, Pseudomonas and Enterobacter. These strains showed dispersed capacities to solubilize P (up to 346 mg L−1) following five days of incubation in NBRIP broth. We also assessed their abilities for indole acetic acid (IAA) production (up to 795,3 µg ml−1) and in vitro salt tolerance. Three Bacillus strains QA1, QA2, and S8 tolerated high salt stress induced by NaCl with a maximum tolerable concentration of 8%. Three performant isolates, QA1, S6 and QF11, were further selected for seed germination assay because of their pronounced abilities in terms of P solubilization, IAA production and salt tolerance. The early plant growth potential of tested strains showed that inoculated quinoa seeds displayed greater germination rate and higher seedlings growth under bacterial treatments. The positive effect on seed germination traits strongly suggests that the tested strains are growth promoting, halotolerant and P solubilizing bacteria which could be exploited as biofertilizers.

Keywords: phosphate solubilizing bacteria, IAA, Seed germination, salt tolerance, quinoa

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609 Prevalence of Mycoplasma hominis and Ureaplasma urealyticum as Causative Agents of Non-Gonococcal Urethritis in Men and Determination of Anti-Bacterial Resistance Rates

Authors: Recep Keşli, Cengiz Demir, Onur Türkyılmaz

Abstract:

Objective: The aim of this study was to determine the prevalence of Mycoplasma hominis and Ureaplasma urealyticum as the causative agents in men with non-gonococcal urethtritis, and anti-bacterial resistance rates. Methods: The Study was carried out in the two Medical Microbiology Laboratories belonging to: Konya Education and Research Hospital and ANS Practice and Research Hospital, Afyon Kocatepe University, between January 2012 and December 2015. Urethral samples were obtained from patients by using a swab. Mycoplasma hominis and Ureaplasma urealyticum were detected by using Mycoplasma IST-2 kit (bio-Mérieux, Marcy l'Étoile, France). Neisseria gonorrhoea was excluded by Gram staining and culture methods. Results: Of all the one hundred and eighty-eight male patients with urethritis, forty M. hominis and forty two U. urealyticum were detected. Resistance rates of M. hominis strains against to doxycycline, ofloxacin, erythromycin, tetracycline, ciprofloxacin, azithromycin, clarithromycin, and pristinamycin were found as 5 %, 65 %, 25 %, 5 %, 80 %, 20 %, 20 %, 20 %, 5 %, respectively. Resistance rates of U. urealyticum strains against to doxycycline, ofloxacin, erythromycin, tetracycline, ciprofloxacin, azithromycin, clarithromycin, and pristinamycin were found as 4.7 %, 66.6 %, 23.8 %, 4.75 %, 81 %, 19 %, 19 %, 4.7 % respectively. No resistance was detected against to josamycin, for both the strains. Conclusions: It was concluded that; ciprofloxacin and ofloxacin had the weakest; josamycin, doxycycline, and tetracycline had the strongest in vitro anti-bacterial activity, for treatment of the NGU. So josamycin, doxycycline, and tetracycline should be preferred as the first choice of anti-bacterial agents, for treatment of the patients with non-gonococcal male urethritis.

Keywords: antimicrobial resistance, Mycoplasma hominis, non-gonococcal urethritis, Ureaplasma urealyticum

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608 Development of 3D Printed, Conductive, Biodegradable Nerve Conduits for Neural Regeneration

Authors: Wei-Chia Huang, Jane Wang

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Damage to nerves is considered one of the most irreversible injuries. The regeneration of nerves has always been an important topic in regenerative medicine. In general, damage to human tissue will naturally repair overtime. However, when the nerves are damaged, healed flesh wound cannot guarantee full restoration to its original function, as truncated nerves are often irreversible. Therefore, the development of treatment methods to successfully guide and accelerate the regeneration of nerves has been highly sought after. In order to induce nerve tissue growth, nerve conduits are commonly used to help reconnect broken nerve bundles to provide protection to the location of the fracture while guiding the growth of the nerve bundles. To prevent the protected tissue from becoming necrotic and to ensure the growth rate, the conduits used are often modified with microstructures or blended with neuron growth factors that may facilitate nerve regeneration. Electrical stimulation is another attempted treatment for medical rehabilitation. With appropriate range of voltages and stimulation frequencies, it has been demonstrated to promote cell proliferation and migration. Biodegradability are critical for medical devices like nerve conduits, while conductive polymers pose great potential toward the differentiation and growth of nerve cells. In this work, biodegradability and conductivity were combined into a novel biodegradable, photocurable, conductive polymer composite materials by embedding conductive nanoparticles in poly(glycerol sebacate) acrylate (PGSA) and 3D-printed into nerve conduits. Rat pheochromocytoma cells and rat neuronal Schwann cells were chosen for the in vitro tests of the conduits and had demonstrate selective growth upon culture in the conductive conduits with built-in microchannels and electrical stimulation.

Keywords: biodegradable polymer, 3d printing, neural regeneration, electrical stimulation

Procedia PDF Downloads 104
607 Chromosomal Damage in Human Lymphocytes by Ultraviolet Radiation

Authors: Felipe Osorio Ospina, Maria Adelaida Mejia Arango, Esteban Onésimo Vallejo Agudelo, Victoria Lucía Dávila Osorio, Natalia Vargas Grisales, Lina María Martínez Sanchez, Camilo Andrés Agudelo Vélez, Ángela Maria Londoño García, Isabel Cristina Ortiz Trujillo

Abstract:

Excessive exposure to ultraviolet radiation, has shown to be a risk factor for photodamage, alteration of the immune mechanisms to recognize malignant cells and cutaneous pro-inflamatorios States and skin cancers. Objective: Identify the time of exposure to ultraviolet radiation for the production of chromosomal damage in human lymphocytes. Methodology: We conducted an in vitro study serial, in which samples were taken from heparinized blood of healthy people, who do not submit exposure to agents that could induce chromosomal alterations. The samples were cultured in RPMI-1640 medium containing 10% fetal bovine serum, penicillin and streptomycin antibiotic. Subsequently, they were grouped and exposed to ultraviolet light for 1 to 20 seconds. At the end of the treatments, cytology samples were prepared, and it was colored with Giemsa (5%). Reading was carried out in an optical microscope and 100 metaphases analysed by treatment for posting chromosomal alterations. Each treatment was conducted at three separate times and each became two replicas. Results: We only presented chromosomal alterations in lymphocytes exposed to UV for a groups 1 to 3 seconds (p<0.05). Conclusions: Exposure to ultraviolet radiation generates visible damage in chromosomes from human lymphocytes observed in light microscopy, the highest rates of injury was observed between two and three seconds, and above this value, the reduction in the number of mitotic cells was evident.

Keywords: ultraviolet rays, lymphocytes, chromosome breakpoints, photodamage

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