Search results for: zoospore inhibition assay

Authors: Forough Ataollahi, Sumit Pramanik, Belinda Pingguan-Murphy, Wan Abu Bakar Bin Wan Abas, Noor Azuan Bin Abu Osman

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Endothelium proliferation is an important process in cardiovascular homeostasis and can be regulated by extracellular environment, as cells can actively sense mechanical environment. In this study, we evaluated endothelial cell proliferation on PDMS/alumina (Al2O3) composites and pure PDMS. The substrates were prepared from pure PDMS and its composites with 5% and 10% Al2O3 at curing temperature 50˚C for 4 h and then characterized by mechanical, structural and morphological analyses. Higher stiffness was found in the composites compared to the pure PDMS substrate. Cell proliferation of the cultured bovine aortic endothelial cells on substrate materials were evaluated via Resazurin assay and 1, 1’-Dioctadecyl-1, 3, 3, 3’, 3’-Tetramethylindocarbocyanine Perchlorate-Acetylated LDL (Dil-Ac-LDL) cell staining, respectively. The results revealed that stiffer substrates promote more endothelial cells proliferation to the less stiff substrates. Therefore, this study firmly hypothesizes that the stiffness elevates endothelial cells proliferation.

Keywords: stiffness, proliferation, bovine aortic endothelial cells, extra cellular matrix, vascular

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Authors: D. Zujur, J. Moret, D. Rodriguez, L. Cruz, J. Lira, L. Gil, E. Dominguez, J. F. Alvarez-Barreto

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In this work, chitosan (CH) has been used to produce a novel coating for Ti6Al4V, the most widely used alloy in orthopedic implants, so as to improve the biological tissue response at the metallic surface. The Ti6Al4V surface was sandblasted with alumina particles and observed by SEM. Chitosan was chemically modified, via crodiimide chemistry, with lactobionic and 4-azidebenzoic acid to make it soluble at physiological pH and photo-crosslinkable, respectively. The reaction was verified by FTIR, NMR, and UV/vis spectroscopy. Ti6Al4V surfaces were coated with solutions of the modified CH and exposed to UV light, causing the polymer crosslinking, and formation of a hydrogel on the surface. The crosslinking reaction was monitored by FTIR at different exposure times. Coating morphology was observed by SEM. The coating´s cytocompatibility was determined in vitro through the culture of rat bone marrow´s mesenchymal stem cells, using an MTT assay. The results show that the developed coating is cytocompatible, easy to apply and could be used for further studies in the encapsulation of bioactive molecules to improve osteogenic potential at the tissue-implant interface.

Keywords: chitosan, photo-crosslinking, Ti6Al4V, bioactive coating, hydrogel

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Authors: Duchel Jeanedvi Kinouani Kinavouidi, Christian Aimé Kayath, Etienne Nguimbi

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Biosurfactants are amphipathic molecules produced by many microorganisms, usually bacteria, fungi, and yeasts. +ey possesses the property of reducing the tension of the membrane interfaces. No studies have been conducted on Shigella species showing the role of biosurfactant-like molecules (BLM) in pathogenicity. +e aim of this study is to assess the ability of Shigella environmental and clinical strains to produce BLM and investigate the involvement of biosurfactants in pathogenicity. Our study has shown that BLM is secreted in the extracellular medium with EI24 ranging from 80% to 100%. +e secretion depends on the type III secretion system (T3SS). Moreover, our results have shown that S. flexneri, S. boydii, and S. sonnei are able to interact with hydrophobic areas with 17.64%, 21.42%, and 22.22% hydrophobicity, respectively. BLM secretion is totally prevented due to the inhibition of T3SS by 100 mM benzoic and 1.5 mg/ml salicylic acids. P. aeruginosa harboring T3SS is able to produce 100% of BLM in the presence or in the absence of both T3SS inhibitors. +e secreted BLM are extractable with an organic solvent such as chloroform, and this could entirely be considered a lipopeptide or polypeptide compound. Secretion of BLM allows some Shigella strains to induce multicellular phenomena like ‘swarming.’

Keywords: shigella flexneri, biosurfactant, T3SS, Lipopeptide

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Authors: Meiling Yu, Nadia Rouatbi, Khuloud T. Al-Jamal

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Background: Treatment of neurological disorders with modern medical and surgical approaches remains difficult. Gene therapy, allowing the delivery of genetic materials that encodes potential therapeutic molecules, represents an attractive option. The treatment of brain diseases with gene therapy requires the gene-editing tool to be delivered efficiently to the central nervous system. In this study, we explored the efficiency of different delivery routes, namely intravenous (i.v.), intra-cranial (i.c.), and intra-nasal (i.n.), to deliver stable nucleic acid-lipid particles (SNALPs) containing gene-editing tools namely Cas9 mRNA and sgRNA encoding for GFP as a reporter protein. We hypothesise that SNALPs can reach the brain and perform gene-editing to different extents depending on the administration route. Intranasal administration (i.n.) offers an attractive and non-invasive way to access the brain circumventing the blood–brain barrier. Successful delivery of gene-editing tools to the brain offers a great opportunity for therapeutic target validation and nucleic acids therapeutics delivery to improve treatment options for a range of neurodegenerative diseases. In this study, we utilised Rosa26-Cas9 knock-in mice, expressing GFP, to study brain distribution and gene-editing efficiency of SNALPs after i.v.; i.c. and i.n. routes of administration. Methods: Single guide RNA (sgRNA) against GFP has been designed and validated by in vitro nuclease assay. SNALPs were formulated and characterised using dynamic light scattering. The encapsulation efficiency of nucleic acids (NA) was measured by RiboGreen™ assay. SNALPs were incubated in serum to assess their ability to protect NA from degradation. Rosa26-Cas9 knock-in mice were i.v., i.n., or i.c. administered with SNALPs to test in vivo gene-editing (GFP knockout) efficiency. SNALPs were given as three doses of 0.64 mg/kg sgGFP following i.v. and i.n. or a single dose of 0.25 mg/kg sgGFP following i.c.. knockout efficiency was assessed after seven days using Sanger Sequencing and Inference of CRISPR Edits (ICE) analysis. In vivo, the biodistribution of DiR labelled SNALPs (SNALPs-DiR) was assessed at 24h post-administration using IVIS Lumina Series III. Results: Serum-stable SNALPs produced were 130-140 nm in diameter with ~90% nucleic acid loading efficiency. SNALPs could reach and stay in the brain for up to 24h following i.v.; i.n. and i.c. administration. Decreasing GFP expression (around 50% after i.v. and i.c. and 20% following i.n.) was confirmed by optical imaging. Despite the small number of mice used, ICE analysis confirmed GFP knockout in mice brains. Additional studies are currently taking place to increase mice numbers. Conclusion: Results confirmed efficient gene knockout achieved by SNALPs in Rosa26-Cas9 knock-in mice expressing GFP following different routes of administrations in the following order i.v.= i.c.> i.n. Each of the administration routes has its pros and cons. The next stages of the project involve assessing gene-editing efficiency in wild-type mice and replacing GFP as a model target with therapeutic target genes implicated in Motor Neuron Disease pathology.

Keywords: CRISPR, nanoparticles, brain diseases, administration routes

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Authors: Piyaporn Buaklang, Narisa Kengtrong Bordeerat

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The use of nanoparticles is increasing worldwide and there are many nanotech-based daily products available in the market. The toxicity of nanoparticles results from their extremely small size which can be transported easily into the blood stream and other organs. We aimed to study the genotoxicity of two nanoparticles, Titanium dioxide (TiO2-NPs) and Zinc oxide (ZnO-NPs), in TK6 cells by micronucleus assay. The cells were tested at 8, 24, and 48 hours after exposed to 0.10, 0.25, 0.50 and 1.00 µg/mL of TiO2-NPs particles size < 25 nm and < 100 nm and to ZnO-NPs at 1, 10, 50, and 100 µg/mL, particles size < 50 nm and < 100 nm. At 24 hours of incubation transmission electron microscope (TEM) revealed that the nanoparticles TiO2-NPs at 1.00 µg/mL and ZnO-NPs at 10 µg/mL were able to be taken into the cells and induced the production of increasing amount of micronucleus in dose-dependent manner. The effect of the two nanoparticles on chromosome aberration indicated that TiO2-NPs and ZnO-NPs are genotoxic. In addition, the toxicity of TiO2-NPs was found to be 10 times more toxic than ZnO-NPs after 24 hours exposure. Analysis showed that the TiO2-NPs induced formation of micronucleus was both time and dose dependent, whereas the genotoxicity of ZnO-NPs was only dose dependent. In conclusion, TiO2-NPs and ZnO-NPs were able to transport through the cells membrane and directly genotoxic to TK6 cells in dose-dependent manner.

Keywords: nanoparticles, genotoxicity, human lymphoblast cells (TK6), micronucleus

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Authors: Rekia. Cherbi, Mokhtar. Saidi, Mohamed. Yousfi, Zhor. Rahmani

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Lawsonia alba (Henna) is widely used in folkloric medicinal for a treatment of various skin diseases such as Eczema (atopic dermatitis), boils and sores. The aim of the present study is to determine the antioxidant activity, total phenolics, flavonoids, and condensed tannins content of extracts from the seeds of Lawsonia. alba grown in Algeria and selected from three different regions (Adrar, Biskra, and Ouargla). Total phenolics content ranged from 68,42 ± 0,54 to 88,31 ± 0,78mg gallic acid equivalents (GAE)/g dry weight, the flavonoids content varied from 1,13 ± 0,0035 to 1,367 ± 0,002mg quercetin equivalents (Q)/ g dry weight and condensed tannins (14,47 ± 0,138 to 25,50 ± 0,076 mg catechin equivalents (CE)/g dry weight). The antioxidant activities of the extracts were evaluated by DPPH assay. The results showed that all extracts from the seeds of Lawsonia. alba seem to be good trappers of radicals, the IC50 values of the extracts ranged between 0,00826 and 0,01 g/l.

Keywords: antioxidant activity, Lawsonia. alba, phenolic compounds, seeds

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Authors: Lyudmila V. Ignatova, Yelena V. Brazhnikova, Togzhan D. Mukasheva, Ramza Zh. Berzhanova, Anel A. Omirbekova

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Endophytic microorganisms are presented in plants of different families growing in the foothills and piedmont plains of Trans-Ili Alatau. It was found that the maximum number of endophytic micromycetes is typical to the Fabaceae family. The number of microscopic fungi in the roots reached (145.9±5.9)×103 CFU/g of plant tissue; yeasts - (79.8±3.5)×102 CFU/g of plant tissue. Basically, endophytic microscopic fungi are typical for underground parts of plants. In contrast, yeasts more infected aboveground parts of plants. Small amount of micromycetes is typical to inflorescence and fruits. Antagonistic activity of selected micromycetes against Fusarium graminearum, Cladosporium sp., Phytophtora infestans and Botrytis cinerea phytopathogens was detected. Strains with a broad, narrow and limited range of action were identified. For further investigations Rh2 and T7 strains were selected, they are characterized by a broad spectrum of fungicidal activity and they formed the large inhibition zones against phytopathogens. Active antagonists are attributed to the Rhodotorula mucilaginosa and Beauveria bassiana species.

Keywords: endophytic micromycetes, fungicidal activity, prevalence, plants

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Authors: Pallavi Moudgil, J. S. Bedi, R. S. Aulakh, J. P. S. Gill

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Mycotoxins in milk are of major public health concern. The present study was envisaged with an aim to monitor the occurrence of aflatoxin M1 and ochratoxin A in raw milk samples collected from individual animals from dairy farms located in Punjab (India). A total of 168 raw milk samples were collected and analysed using competitive ELISA kits. Out of these, 9 (5.4%) samples were found positive for aflatoxin M1 with the mean concentration of 0.006-0.13 ng/ml and 2 (1.2%) samples exceeded the established maximum residue limit of 0.05 ng/ml established by the European Union. For ochratoxin A, 2 (0.1%) samples were found positive with the mean concentration of 0.61-0.83 ng/ml with both the samples below the established maximum residue limit of 2 ng/ml. The results showed that the milk of dairy cattle is safe with respect to ochratoxin A contamination but occurrence of aflatoxin M1 above maximum residue limit suggested that feed contaminated with mycotoxins might have been offered to dairy cattle that can pose serious health risks to consumers.

Keywords: Aflatoxin M1, health risks, maximum residue limit, milk, Ochratoxin A

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Authors: Li Yuchen, Wu Qingxin, Jin Yuxing, Yang Qian

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Interleukin-11 (IL-11), a well-known anti-inflammatory factor, helps to protect against intestinal epithelium damage caused by physical or chemical factors. However, little is known about the role of IL-11 during viral infection. Herein, high mRNA and protein levels of IL-11 were found in epithelial cells and jejunum of piglets during porcine epidemic diarrhea virus (PEDV) infection, and IL-11 expression was positively correlated with the level of viral infection. Pretreatment with recombinant porcine IL-11 (pIL-11) suppressed PEDV replication in Vero E6 cells, while IL-11 knockdown promoted viral infection. Furthermore, pIL-11 inhibited viral infection by preventing PEDV-mediated apoptosis of cells through activating the IL-11/STAT3 signal pathway. Conversely, application of a STAT3 phosphorylation inhibitor significantly antagonized the anti-apoptosis function of pIL-11 and counteracted its inhibition of PEDV. Our data suggested that that IL-11 is a novel PEDV-inducible cytokine, and its production enhances the anti-apoptosis ability of epithelial cells against PEDV infection. The potential uses of IL-11 as a novel therapeutic against devastating viral diarrhea in piglets deserves more attention and study.

Keywords: Interleukin-11, Porcine epidemic diarrhea virus, STAT3, anti-apoptosis

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Authors: Abubakar Umar Birnin-yauri

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Plastic wastes arising from Polyethylene (PE)-based materials are increasingly becoming environmental problem, this is owed to the fact that these PE waste materials will only decompose over hundreds, or even thousands of years, during which they cause serious environmental problems. In this research, Polymer blends prepared from PE, Cashewnut flour (CNF) and Gum Arabic (GA) were studied in order to assay their biodegradation potentials via composting method. Different sample formulations were made i.e., X1= (70% PE, 25% CNF and 5% GA, X2= (70% PE, 20% CNF and 10% GA), X3= (70% PE, 15% CNF and 15% GA), X4 = (70% PE, 10% CNF and 20% GA) and X5 = (70% PE, 5% CNF and 25% GA) respectively. The results obtained showed that X1 recorded weight loss of 9.89% of its original weight after the first 20 days and 37.45% after 100 day, and X2 lost 12.67 % after the first 20 days and 42.56% after 100day, sample X5 experienced the greatest weight lost in the two methods adopted which are 52.9% and 57.89%. Instrumental analysis such as Fourier Transform Infrared Spectroscopy, Thermogravimetric analysis and Scanning electron microscopy were performed on the polymer blends before and after biodegradation. The study revealed that the biodegradation of the polymer blends is influenced by the contents of both the CNF and GA added into the blends.

Keywords: polyethylene, cashewnut, gum Arabic, biodegradation, blend, environment

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Authors: A. Jamsheera, F. Arjmand, D. K. Mohapatra

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Small molecules binding to specific sites along DNA molecule are considered as potential chemotherapeutic agents. Their role as mediators of key biological functions and their unique intrinsic properties make them particularly attractive therapeutic agents. Keeping in view, novel dipeptide complexes Cu(II)-Val-Pro (1), Zn(II)-Val-Pro (2), Cu(II)-Ala-Pro (3) and Zn(II)-Ala-Pro (4) were synthesized and thoroughly characterized using different spectroscopic techniques including elemental analyses, IR, NMR, ESI–MS and molar conductance measurements. The solution stability study carried out by UV–vis absorption titration over a broad range of pH proved the stability of the complexes in solution. In vitro DNA binding studies of complexes 1–4 carried out employing absorption, fluorescence, circular dichroism and viscometric studies revealed the binding of complexes to DNA via groove binding. UV–vis titrations of 1–4 with mononucleotides of interest viz., 5´-GMP and 5´-TMP were also carried out. The DNA cleavage activity of the complexes 1 and 2 were ascertained by gel electrophoresis assay which revealed that the complexes are good DNA cleavage agents and the cleavage mechanism involved a hydrolytic pathway. Furthermore, in vitro antitumor activity of complex 1 was screened against human cancer cell lines of different histological origin.

Keywords: dipeptide Cu(II) and Zn(II) complexes, DNA binding profile, pBR322 DNA cleavage, in vitro anticancer activity

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Authors: E. Carraro, Sa. Bonetta, Si. Bonetta, E. Ceretti, G. C. V. Viola, C. Pignata, S. Levorato, T. Salvatori, S. Vannini, V. Romanazzi, A. Carducci, G. Donzelli, T. Schilirò, A. De Donno, T. Grassi, S. Bonizzoni, A. Bonetti, G. Gilli, U. Gelatti

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In 2013, air pollution and particulate matter were classified as carcinogenic to human by the IARC. At present, PM is Europe's most problematic pollutant in terms of harm to health, as reported by European Environmental Agency (EEA) in the EEA Technical Report on Air quality in Europe, 2015. A percentage between 17-30 of the EU urban population lives in areas where the EU air quality 24-hour limit value for PM10 is exceeded. Many studies have found a consistent association between exposure to PM and the incidence and mortality for some chronic diseases (i.e. lung cancer, cardiovascular diseases). Among the mechanisms responsible for these adverse effects, genotoxic damage is of particular concern. Children are a high-risk group in terms of the health effects of air pollution and early exposure during childhood can increase the risk of developing chronic diseases in adulthood. The MAPEC_LIFE (Monitoring Air Pollution Effects on Children for supporting public health policy) is a project founded by EU Life+ Programme (LIFE12 ENV/IT/000614) which intends to evaluate the associations between air pollution and early biological effects in children and to propose a model for estimating the global risk of early biological effects due to air pollutants and other factors in children. This work is focused on the micronuclei frequency in child buccal cells in association with airborne PM levels taking into account the influence of other factors associated with the lifestyle of children. The micronucleus test was performed in exfoliated buccal cells of 6–8 years old children from 5 Italian towns with different air pollution levels. Data on air quality during the study period were obtained from the Regional Agency for Environmental Protection. A questionnaire administered to children’s parents was used to obtain details on family socio-economic status, children health condition, exposures to other indoor and outdoor pollutants (i.e. passive smoke) and life-style, with particular reference to eating habits. During the first sampling campaign (winter 2014-15) 1315 children were recruited and sampled for Micronuclei test in buccal cells. In the sampling period the levels of the main pollutants and PM10 were, as expected, higher in the North of Italy (PM10 mean values 62 μg/m3 in Torino and 40 μg/m3 in Brescia) than in the other towns (Pisa, Perugia, Lecce). A higher Micronucleus frequency in buccal cells of children was found in Brescia (0.6/1000 cells) than in the other towns (range 0.3-0.5/1000 cells). The statistical analysis underlines a relation of the micronuclei frequency with PM concentrations, traffic level near child residence, and level of education of parents. The results suggest that, in addition to air pollution exposure, some other factors, related to lifestyle or further exposures, may influence micronucleus frequency and cellular response to air pollutants.

Keywords: air pollution, buccal cells, children, micronucleus cytome assay

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Authors: Laila H. Abdel-Rahman, Mohamed Shaker S. Adam, Ahmed M. Abu-Dief, Hanan El-Sayed Ahmed

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In this investigation, Cu(II), Pd(II) and Ag(I) complexes with the tetra-dentate DSPH Schiff base ligand were synthesized. The DSPH Schiff base and its complexes were characterized by using different physicochemical and spectral analysis. The results revealed that the metal ions coordinated with DSPH ligand through azomethine nitrogen and phenolic oxygen. Cu(II), Pd(II) and Ag(I) complexes are present in a 1:1 molar ratio. Pd(II) and Ag(I) complexes have square planar geometries while, Cu(II) has a distorted octahedral (Oh) geometry. All investigated complexes are nonelectrolytes. The investigated compounds were tested against different strains of bacteria and fungi. Both prepared compounds showed good results of inhibition against the selected pathogenic microorganism. Moreover, the interaction of investigated complexes with CT-DNA was studied via various techniques and the binding modes are mainly intercalative and grooving modes. Operating Environment MOE package was used to do docking studies for the investigated complexes to explore the potential binding mode and energy. Furthermore, the growth inhibitory effect of the investigated compounds was examined on some cancer cells lines.

Keywords: tetradentate, antimicrobial, CT-DNA interaction, docking, anticancer

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Authors: Maomao Cao

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Objective: To test the performance of ctDNA methylation for the detection of liver cancer. Methods: A total of 1233 individuals have been recruited in 2017. 15 male and 15 female samples (including 10 cases of liver cancer) were randomly selected in the present study. CfDNA was extracted by MagPure Circulating DNA Maxi Kit. The concentration of cfDNA was obtained by Qubit™ dsDNA HS Assay Kit. A pre-constructed predictive model was used to analyze methylation data and to give a predictive score for each cfDNA sample. Individuals with a predictive score greater than or equal to 80 were classified as having liver cancer. CT tests were considered the gold standard. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) for the diagnosis of liver cancer were calculated. Results: 9 patients were diagnosed with liver cancer according to the prediction model (with high sensitivity and threshold of 80 points), with scores of 99.2, 91.9, 96.6, 92.4, 91.3, 92.5, 96.8, 91.1, and 92.2, respectively. The sensitivity, specificity, positive predictive value, and negative predictive value of ctDNA methylation for the diagnosis of liver cancer were 0.70, 0.90, 0.78, and 0.86, respectively. Conclusions: ctDNA methylation could be an acceptable diagnostic modality for the detection of liver cancer.

Keywords: liver cancer, ctDNA methylation, detection, diagnostic performance

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Authors: Abdolreza Esmaeilzadeh, Mehri Mirzaei

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Objective: Rheumatic disease is a chronic disease that causes pain, stiffness, swelling and limited motion and function of many joints. RA is the most common form of autoimmune arthritis, affecting more than 1.3 million Americans. Of these, about 75% are women. Materials and Methods: This study was formed due to the misconception about ANA test, which is frequently performed with methods based upon solid phase as ELISA. This experiment was conducted on 430 patients, with clinical symptoms that are likely affected with rheumatic diseases, simultaneously by means of ANA and FANA. Results: 36 cases (8.37%) of patients, despite positive ANA, have demonstrated negative results via Indirect Immunofluorescence Assay (IIFA), (false positive). 116 cases (27%) have demonstrated negative ANA results, by means of the ELISA technique, although they had positive IIFA results. Conclusion: Other advantages of IIFA are antibody titration and specific pattern detection that have the capability of distinguishing positive dsDNA results. According to the restrictions and false negative cases, in patients, IIFA test is highly recommended for these disease's diagnosis.

Keywords: autoimmune disease, IIFA, EIA, rheumatic disease

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Authors: Sunil Kamboj, Vipin Saini, Suman Bala, Gaurav Sharma

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The main aim of the present research was to encapsulate mefenamic acid in niosomes and incorporate the prepared niosomes in the carbopol gel base for sustained therapeutic action. Mefenamic acid loaded niosomes were prepared by thin film hydration technique and evaluated for entrapment efficiency, vesicular size and zeta potential. The entrapment efficiency of the prepared niosomes was found to increase with decreasing the HLB values of surfactants and vesicle size was found to increase with increasing the cholesterol concentration. Niosomal vesicles with good entrapment efficiencies were incorporated in carbopol gel base to form the niosomal gel. The prepared niosomal gel was evaluated for pH, viscosity, spreadability, extrudability and skin permeation study across the rat skin.The results of permeation study revealed that the gel formulated with span 60 niosomes sustained the drug release for 12 h. Further the in vivo study showed the good inhibition of inflammation by the gel prepared with span 60 niosomes.

Keywords: mefenamic acid, niosomal gel, nonionic surfactants, sustained release

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Authors: Zhongyang Sun, Shu Zhang

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In osteoblasts, L-type voltage sensitive calcium channels (LTCCs), especially the Cav1.2 LTCCs, play fundamental roles in cellular responses to external stimuli including both mechanical forces and hormonal signals. Several lines of evidence have revealed that the density of bone is increased and the resorption of bone is decreased when these calcium channels in osteoblasts are activated. And numerous studies have shown that mechanical loading promotes bone formation in the modeling skeleton, whereas removal of this stimulus in microgravity results in a reduction in bone mass. However, the effect of microgravity on LTCCs in osteoblasts is still unknown. The aim of this study was to determine whether microgravity exerts influence on LTCCs in osteoblasts and the possible mechanisms underlying. In this study, we demonstrate that simulated microgravity substantially inhibits LTCCs in osteoblast by suppressing the expression of Cav1.2. Then we show that the up-regulation of miR-103 is involved in the down-regulation of Cav1.2 expression and inhibition of LTCCs by simulated microgravity in osteoblasts. Our study provides a novel mechanism of simulated microgravity-induced adverse effects on osteoblasts, offering a new avenue to further investigate the bone loss caused by microgravity.

Keywords: L-type voltage sensitive calcium channels, Cav1.2, osteoblasts, microgravity

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Authors: N. Mezıou-Cheboutı, A. Merabet, N. Behidj, F. Z. Bissaad

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The Algeria by its location, offers a rich and diverse vegetation. A large number of aromatic and medicinal plants grow spontaneously. The interest in these plants has continued to grow in recent years. Their particular properties due to the essential oil fraction can be utilized to treat microbial infections. To this end, and in the context of the valuation of the Algerian flora, we became interested in the species of the family Lamiaceae which is one of the most used as a global source of spices and extracts strong families antimicrobial potency. The plant on which we have based our choice is a species of sage "Salvia officinalis" from the Isser localized region within the province of Boumerdes. This work focuses on the study of the antimicrobial activity of essential oil extracted from the leaves of salvia officinalis. The extraction is carried out by HE hydrodistillation and reveals a yield of 1.06℅. The study of the antimicrobial activity of the essential oil by the method of at aromatogramme shown that Gram positive bacteria are most susceptible (Staphylococcus aureus and Bacillus subtilis) with a strong inhibition of growth. The yeast Candida albicans fungus Aspergillus niger and have shown moderately sensitive.

Keywords: Salvia officinalis, steam distillation, essential oil, aromatogram, anti-microbial activity

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Authors: Josselyn Mata Calidonio, Arianna I. Maddox, Kimberly Hamad-Schifferli

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Rapid diagnostics are critical infectious disease tools that are designed to detect a known biomarker using antibodies specific to that biomarker. However, a way to detect unknown viruses has not yet been achieved in a paper test format. We describe here a route to make an adaptable paper immunoassay that can detect an unknown biomarker, demonstrating it on SARS-CoV-2 variants. The immunoassay repurposes cross-reactive antibodies raised against the alpha variant. Gold nanoparticles of two different colors conjugated to two different antibodies create a colorimetric signal, and machine learning of the resulting colorimetric pattern is used to train the assay to discriminate between variants of alpha and Omicron BA.5. By using principal component analysis, the colorimetric test patterns can pick up and discriminate an unknown that it has not encountered before, Omicron BA.1. The test has an accuracy of 100% and a potential calculated discriminatory power of 900. We show that it can be used adaptively and that it can be used to pick up emerging variants without the need to raise new antibodies.

Keywords: adaptive immunoassay, detecting unknown viruses, gold nanoparticles, paper immunoassay, repurposing antibodies

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Authors: Abdelsabour G. A. Khaled, Ahmed W. A. Abdalla And Sabry Y. M. Mahmoud

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Banana plants showing typical mosaic and yellow stripes on leaves as symptoms were collected from Assiut Governorate in Egypt. The causal agent was identified as Cucumber mosaic virus (CMV) on the basis of symptoms, transmission, serology, transmission electron microscopy and reverse transcription polymerase chain reaction (RT-PCR). Coat protein (CP) gene was amplified using gene specific primers for coat protein (CP), followed by cloning into desired cloning vector for sequencing. In this study the CMV was transmitted into propagation host either by aphid or mechanically. The transmission was confirmed through Direct Antigen Coating Enzyme Linked Immuno Sorbent Assay (DAC-ELISA). Analysis of the 120 deduced amino acid sequence of the coat protein gene revealed that the EG-A strain of CMV shared from 97.50 to 98.33% with those strains belonging to subgroup IA. The cluster analysis grouped the Egyptian isolate with strains Fny and Ri8 belonging sub-group IA. It appears that there occurs a high incidence of CMV infecting banana belonging to IA subgroup in most parts of Egypt.

Keywords: banana, CMV, transmission, CP gene, RT-PCR

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Authors: Chijindu Okpalaoka, Charles Chinedu Onuselogu

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COVID-19, humanity's coronavirus disease caused by SARS-CoV-2, was first detected in late 2019 in Wuhan, China. COVID-19 lacked an established conventional pharmaceutical therapy, and as a result, the outbreak quickly became an epidemic affecting the entire World. Only a qPCR assay is reliable for diagnosing COVID-19. Clustered, regularly interspaced short palindromic repeats (CRISPR) technology is being researched for speedy and specific identification of COVID-19, among other therapeutic techniques. Apart from its therapeutic capabilities, the CRISPR technique is being evaluated to develop antiviral therapies; nevertheless, no CRISPR-based medication has been approved for human use to date. Prophylactic antiviral CRISPR in living being cells, a Cas 13-based approach against coronavirus, has been developed. While this method can be evolved into a treatment approach, it may face substantial obstacles in human clinical trials for licensure. This study discussed the potential applications of CRISPR-based techniques for developing a speedy and accurate feasible treatment alternative for the COVID-19 virus.

Keywords: COVID-19, CRISPR technique, Cas13, SARS-CoV-2, prophylactic antiviral

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Authors: Abolfazl Kamkar

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Vegetable oils and fats are recognized as important components of our diet. They provide essential fatty acids, which are precursors of important hormones and control many physiological factors such as blood pressure, cholesterol level, and the reproductive system.Vegetable oils with higher contents of unsaturated fatty acids, especially polyunsaturated fatty acids (PUFAs) are more susceptible to oxidation.Protective effects of Sature jahortensis(SE) extracts in stabilizing soybean oil at different concentrations (200 and 400 ppm) were tested. Results showed that plant extracts could significantly (P< 0.05) lower the peroxide value and thiobarbituric acid value of oil during storage at 60 oC. The IC50 values for methanol and ethanol extracts were 31.5 ± 0.7 and 37.00 ± 0 µg/ml, respectively. In the β- carotene/linoleic acid system, methanol and ethanol extracts exhibited 87.5 ± 1.41% and 74.0 ±2.25 % inhibition against linoleic acid oxidation. The total phenolic and flavonoid contents of methanol and ethanol extracts were (101.58 ± 0. 26m g/ g) and (96.00 ± 0.027 mg/ g), (44.91 ± 0.14 m g/ g) and (14.30 ± 0.12 mg/ g) expressed in Gallic acid and Quercetin equivalents, respectively.These findings suggest that Satureja extracts may have potential application as natural antioxidants in the edible oil and food industry.

Keywords: satureja hortensis, antioxidant activity, oxidative stability, vegetable oil, extract

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Authors: Reza Sadeghi

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In this study, microwave heating was employed for controlling Oryzaephilus mercator. adults infesting stored Iranian dried fig. For this purpose, the dried fig samples were artificially infested with O. mercator and then heated in a microwave oven (2450 MHz) at the power outputs of 450, 720, and 900 W for 10, 20, 30, and 40 s, respectively. Subsequently, changes in the colors of the product samples under the effects of the varied microwave applications were investigated in terms of lightness (ΔL*), redness (Δa*), and yellowness (Δb*) using an image processing technique. The results revealed that both parameters of microwave power and exposure time had significant impacts on the pest mortality rates (p<0.01). In fact, a direct positive relationship was obtained between the mortality rate and microwave irradiation power. Complete mortality was achieved for the pest at the power of 900 W and exposure time of 40 s. The dried fig samples experienced fewer changes in their color parameters. Considering the successful pest control and acceptable changes in the product quality, microwave irradiation can be introduced as an appropriate alternative to chemical fumigants.

Keywords: colorimetric assay, microwave heating, Oryzaephilus mercator, mortality

Procedia PDF Downloads 89

Authors: E. Kuley, M. Durmuş, E. Balikci, G. Ozyurt, Y. Uçar, F. Kuley, F. Ozogul, Y. Ozogul

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Ammonia and biogenic amine production of fish spoilage bacteria in sardine infusion decarboxylase broth and antimicrobial effect of olive by products (olive leaf extract:OL, olive cake: OC and black water:BW) was monitored using HPLC method. Fish spoilage bacteria produced all biogenic amine tested, mainly histamine and serotonin. Ammonia was accumulated more than 13.60 mg/L. Histamine production was in range 37.50 mg/L by Ser. liquefaciens and 86.71 mg/L by Ent. cloacae. The highest putrescine and cadaverine production was observed by Ent. cloacae (17.80 vs. 17.69 mg/L). The presence of OL, OC and BW in the broth significantly affected biogenic amine accumulation by bacteria. The antibacterial effect of olive by products depended on bacterial strains. OL and OC resulted in significant inhibition effect on HIS accumulation by bacteria apart from Ser. liquefaciens and Prot. mirabilis. The study result revealed that usefulness of OL and OC to prevent the accumulation of this amine which may affect human health.

Keywords: Antimicrobials, biogenic amine, fish spoilage bacteria, olive-by products

Procedia PDF Downloads 503

Authors: Antonella Di Sotto, Cinzia Ingallina, Caterina Fraschetti, Simone Circi, Marcello Locatelli, Simone Carradori, Gabriela Mazzanti, Luisa Mannina, Silvia Di Giacomo

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Cannabis sativa L. is one of the oldest cultivated plant species known not only for its voluptuous use but also for the wide application in food, textile, and therapeutic industries. Recently, the progress of biotechnologies applied to medicinal plants has allowed to produce different hemp varieties with low content of psychotropic phytoconstituents (tetrahydrocannabinol < 0.2% w/v), thus leading to a renewed industrial and therapeutic interest for this plant. In this context, in order to discover new potential remedies of pharmaceutical and/or nutraceutical interest, the chemopreventive properties of different organic and hydroalcoholic extracts, obtained from the inflorescences of C. sativa L. var. USO31, collected in June and September harvesting, were assessed. Particularly, the antimutagenic activity towards the oxidative DNA-damage induced by tert-butyl hydroperoxide (t-BOOH) was evaluated, and the DPPH (2,2-diphenyl-1-picrylhydrazyl) and ABTS (2,2'-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid) radical scavenging power of the samples were assessed as possible mechanisms of antimutagenicity. Furthermore, the ability of the extracts to inhibit the glucose-6-phosphate dehydrogenase (G6PD), whose overexpression has been found to play a critical role in neoplastic transformation and tumor progression, has been studied as a possible chemopreventive strategy. A careful phytochemical characterization of the extracts for phenolic and terpenoid composition has been obtained by high performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC-MS) methods. Under our experimental condition, all the extracts were found able to interfere with the tBOOH-induced mutagenicity in WP2uvrAR strain, although with different potency and effectiveness. The organic extracts from both the harvesting periods were found to be the main effective antimutagenic samples, reaching about a 55% inhibition of the tBOOH-mutagenicity at the highest concentration tested (250 μg/ml). All the extracts exhibited radical scavenger activity against DPPH and ABTS radicals, with a higher potency of the hydroalcoholic samples. The organic extracts were also able to inhibit the G6PD enzyme, being the samples from September harvesting the highly potent (about 50% inhibition respect to the vehicle). At the phytochemical analysis, all the extracts resulted to contain both polar and apolar phenolic compounds. The HPLC analysis revealed the presence of catechin and rutin as the major constituents of the hydroalcoholic extracts, with lower levels of quercetin and ferulic acid. The monoterpene carvacrol was found to be an ubiquitarian constituent. At GC-MS analysis, different terpenoids, among which caryophyllene sesquiterpenes, were identified. This evidence suggests a possible role of both polyphenols and terpenoids in the chemopreventive properties of the extracts from the inflorescences of C. sativa var. USO31. According to the literature, carvacrol and caryophyllene sesquiterpenes can contribute to the strong antimutagenicity although the role of all the hemp phytocomplex cannot be excluded. In conclusion, present results highlight a possible interest for the inflorescences of C. sativa var. USO31 as source of bioactive molecules and stimulate further studies in order to characterize its possible application for nutraceutical and pharmaceutical purposes.

Keywords: antimutagenicity, glucose-6-phosphate dehydrogenase, hemp inflorescences, nutraceuticals, sesquiterpenes

Procedia PDF Downloads 158

Authors: Sajjad Negahban, Ruihe Wang, Baojiang Sun

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Gas-lift dual gradient drilling is a solution for deepwater drilling challenges. As well, Continuous development of drilling technology leads to increase employment of mineral oil based drilling fluids and synthetic-based drilling fluids, which have adequate characteristics such as: high rate of penetration, lubricity, shale inhibition and low toxicity. The paper discusses utilization of nitrified mineral oil base drilling for deepwater drilling and for more accurate prediction of pressure in DGD at marine riser, solubility and bubble pressure were considered in steady state hydraulic model. The Standing bubble pressure and solubility correlations, and two models which were acquired from experimental determination were applied in hydraulic model. The effect of the black oil correlations, and new solubility and bubble pressure models was evaluated on the PVT parameters such as oil formation volume factor, density, viscosity, volumetric flow rate. Eventually, the consequent simulated pressure profile due to these models was presented.

Keywords: solubility, bubble pressure, gas-lift dual gradient drilling, steady state hydraulic model

Procedia PDF Downloads 278

Authors: S. Ghalem, M. Mesmoudi, I. Daoudand, H. Allali

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The nitrogen-containing bisphosphonates (N-BPs) are well established as the treatments of choice for disorders of excessive bone resorption, myeloma and bone metastases, and osteoporosis. They inhibit farnesyl pyrophosphate synthase (FFPS), a key enzyme in the mevalonate pathway, resulting in inhibition of the prenylation of small GTP-binding proteins in osteoclasts and disruption of their cytoskeleton, adhesion/spreading, and invasion of cancer cells. A very few examples for synthesis of α-amino bisphosphonates based on several amino acids are known from the literature. In the present work, esters of aminoacid react with ketophsophonate (or their analog acid or acyl) to afford the desired products, α-iminophosphonates. The reaction of imine with dimethyl phosphate in the presence of catalytic amount of I2 give ester of α-aminobisphosphonate as sole product in good yield. Finally, we used computational docking methods to predict how several α-aminobisphosphonates bind to FPPS and how R and X influence. Pamidronate, β-aminobisphosphonate already marketed, was used as reference. These results are of interest since they represent a new and simple way to sythesize α-aminobisphosphonates with a free COOH group increased by R2 functionalisable and opening up the possibility of using the molecular docking to facilitate the design of other, novel FFPS inhibitors.

Keywords: drug research, cancer, α-amino bisphosphonates, molecular docking

Procedia PDF Downloads 271

Authors: Kwanchai Ratanamanee, Pattra Ahmadi Pirshahid, Yaowaluk Khamphan, Sirinan Thubthimthad

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Obesity is a main global health problem. The obesity rated has continued to be higher and higher. It causes to serious systems, diabetes, coronary artery disease, stroke, and some types of cancer. Oristat is one of the best drugs worldwide used as a pancreatic lipase inhibitor. To develop the new therapeutic drugs from medicinal plant always explored. In this study, 24 medicinal plants were investigated for their pancreatic lipase and cholesterol esterase inhibitory effects with Fluorometer assay and oristat as a positive control. It showed that the ethanolic extract of pods of Acacia concinna (Willd.) D.C., possess pancreatic lipase and cholesterol esterase inhibitory activities of IC50 at 2.73 and 3.77 mg/ml respectively as well as oral acute toxicity of the extract (LD50) was 6,300 mg/kg body weight. The extract of A.concinna should be further investigated in animal testing. The results of pancreatic lipase and cholesterol esterase inhibitor of the extracts will lead us to utilize A.concinna for developing as obesity dietary supplement from a medicinal plant.

Keywords: Acacia concinna (Willd.) D. C., cholesterol esterase, obesity, pancreatic lipase

Procedia PDF Downloads 481

Authors: Jamie A. Prowse Turner, Jamie I. D. Campbell, Valerie A. Thompson

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The objective of the current research was to use a dual-process theory framework to explain these age-related differences in reasoning. Seventy-two older (M = 80.0 years) and 72 younger (M = 24.6 years) adults were given a variety of reasoning tests (i.e., a syllogistic task, base rate task, the Cognitive Reflection Test, and a perspective manipulation), as well as independent tests of capacity (working memory, processing speed, and inhibition), thinking styles, and metacognitive ability, to account for these age-related differences. It was revealed that age-related differences were limited to problems that required Type 2 processing and were related to differences in cognitive capacity, individual difference factors, and strategy choice. Furthermore, older adults’ performance can be improved by reasoning from another’s’ perspective and cannot, at this time, be explained by metacognitive differences between young and older adults. All of these findings fit well within a dual-process theory of reasoning, which provides an integrative framework accounting for previous findings and the findings presented in the current manuscript.

Keywords: aging, dual-process theory, performance, reasoning ability

Procedia PDF Downloads 192

Authors: Benarous Khedidja, Yousfi Mohamed

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Infections caused by Candida species manifest in a number of diseases, including candidemia, vulvovaginal candidiasis, endocarditis, and peritonitis. These Candida species have been reported to have lipolytic activity by secretion of lipolytic enzymes such as esterases, lipases and phospholipases. These Extracellular hydrolytic enzymes seem to play an important role in Candida overgrowth. Candidiasis is commonly treated with antimycotics such as clotrimazole and nystatin, which bind to a major component of the fungal cell membrane (ergosterol). This binding forms pores in the membrane that lead to death of the fungus. Due to their secondary effects, scientists have thought of another treatment basing on lipase inhibition but we haven’t found any lipase inhibitors used as candidiasis treatment. In this work, we are interested to lipases inhibitors such as alkaloids as another candidiasis treatment. In the first part, we have proceeded to optimize the alkaloid structures and protein 3D structure using Hyperchem software. Secondly, we have docked inhibitors using Genetic algorithm with GOLD software. The results have shown ten possibilities of binding inhibitor to Candida rugosa lipase (CRL) but only one possibility has been accepted depending on the weakest binding energy.

Keywords: marrubiin, candida rugosa lipase, docking, gold

Procedia PDF Downloads 246