Search results for: zoospore inhibition assay

Authors: Purnima Rawat, Divya Vohora, Sarika Gupta, Farhan J. Ahmad, Sushama Talegaonkar

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Nanoparticles (NPs) that target bone tissue were developed using PLGA–mPEG (poly(lactic-co-glycolic-acid)–polyethylene glycol) diblock copolymers by using pamidronate as a bone-targeting moieties. These NPs are expected to enable the transport of hydrophilic drugs. The NP was prepared by in situ polymerization method, and their in- vitro characteristics were evaluated using dynamic light scattering, transmission electron microscopy (TEM) and in phosphate-buffered solution. The bone targeting potential of the NP was also evaluated on in-vitro pre-osteoblast MCT3E1 cell line using ALP activity, degree of mineralization and RT-PCR assay. The average particle size of the NP was 101.6 ± 3.7nm, zeta potential values were negative (-25±0.34mV) of the formulations and the entrapment efficiency was 93± 3.1 % obtained. The moiety of the PLGA–mPEG–pamidronate NPs exhibited the best apatite mineral binding ability in-vitro MCT3E1 pre-osteoblast cell line. Our results suggested that the developed nanoparticles may use as a delivery system for Pamidronate in bone repair and regeneration, warranting further evaluation of the treatment of bone disease.

Keywords: nanoparticle, pamidronate, in-situ polymerization, osteoblast

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Authors: Ashish Kumar Jain, Deepak Mishra

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The objective of the present investigation was to prepare and evaluate a vesicular dual drug delivery system for effective management of mucosal ulcer. Inner encapsulating and Double liposomes were prepared by glass bead and reverse phase evaporation method respectively. The formulation consisted of inner liposomes bearing Ranitidine Bismuth Citrate (RBC) and outer liposomes encapsulating Amoxicillin trihydrate (AMOX). The optimized inner liposomes and double liposomes were extensively characterized for vesicle size, morphology, zeta potential, vesicles count, entrapment efficiency and in vitro drug release. In vitro, the double liposomes demonstrated a sustained release of AMOX and RBC viz 91.4±1.8% and 77.2±2.1% respectively at the end of 72 hr. Furthermore binding specificity and targeting propensity toward H. pylori (SKP-56) was confirmed by agglutination and in situ adherence assay. Reduction of the absolute alcohol induced ulcerogenic index from 3.01 ± 0.25 to 0.31 ± 0.09 and 100% H. pylori clearance rate was observed. These results suggested that double liposomes are potential vector for the development of dual drug delivery for effective treatment of H. pylori-associated peptic ulcer.

Keywords: double liposomes, H. pylori targeting, PE liposomes, glass-beads method, peptic ulcers

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Authors: Jyothi Nagraj, Sudeshna Mukherjee, Rajdeep Chowdhury

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Autophagy has recently been linked with cancer cell survival post drug insult contributing to acquisition of resistance. However, the molecular signaling governing autophagic survival response is poorly explored. In our study, in osteosarcoma (OS) cells cisplatin shock was found to activate both MAPK and autophagy signaling. An activation of JNK and autophagy acted as pro-survival strategy, while ERK1/2 triggered apoptotic signals upon cisplatin stress. An increased sensitivity of the cells to cisplatin was obtained with simultaneous inhibition of both autophagy and JNK pathway. Furthermore, we observed that the autophagic stimulation upon drug stress regulates other developmentally active signaling pathways like the Hippo pathway in OS cells. Cisplatin resistant cells were thereafter developed by repetitive drug exposure followed by clonal selection. Basal levels of autophagy were found to be high in resistant cells to. However, the signaling mechanism leading to autophagic up-regulation and its regulatory effect differed in OS cells upon attaining drug resistance. Our results provide valuable clues to regulatory dynamics of autophagy that can be considered for development of improved therapeutic strategy against resistant type cancers.

Keywords: JNK, autophagy, drug resistance, cancer

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Authors: Meziani Samira, Menadi Noreddine, Labga Lahouaria, Chenni Fatima Zohra, Toumi Asma

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A comparative study between two varieties of soya beans was carried out in this work. The method consists of studying and proceeding to prepare a by-product (Flour) from two varieties of soybeans, a Chinese variety imported and marketed in Algeria. The chemical composition of ash, protein and fat was determined in this study. The minerals, namely potassium and sodium, were measured by flame spectrophotometer. In addition, the estimation of the polyphenol content and evaluation of the antioxidant activity Ferric Reducing Antioxidant Power assay (FRAP) f the methanol extracts of the flours were also carried out. The result revealed that soy flour from two cultivars, on average, contained 8% moisture, more than 50% protein, 1.58-1.87g fat, and 0.28-0.30g of ash. A slight difference was found for contents of 489 mg/ml of K + and 20 mg/ml of NA +. In addition, the phenolic content of the methanolic extracts gives a value of almost 37 mg EAG / g for both cultivars of soy flour. The estimated Reductive Antioxidant Iron (FRAP) potency of soy flour might be related to its polyphenol richness, which is similar to the variety of China. The flour Soya varieties tested contained a significant amount of protein and phenolic compounds with good antioxidant properties.

Keywords: soye beans, soya flour, protein, total polyphenols

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Authors: Abdelkrim Cheriti, Mebarka Belboukhari, Nasser Belboukhari

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Launaea Cass. is a small genus of the family Asteraceae (tribe Lactuceae, subtribe Sonchinae), consisting of 54 species, of which 9 are presented in the flora of Algeria and is mainly distributed in the South Mediterranean, Africa and SW Asia. Plants in the Launaea genus have been used ethnobotanically as bitter stomachic, for treating diarrhea, gastrointestinal tracts, as anti-inflammatory, for skin diseases, treatment of infected wounds, hepatic pains, children fever, as soporific, lactagogue, diuretic and as insecticidal. Antioxidants are vital substances, which possess the ability to protect the body from damages caused by free radical induced oxidative stress. A variety of free radical scavenging antioxidants is found in a number of dietary sources. The main objective of this study focused on the screening of antioxidant activity of Launaea nudicaulis (Asteraceae) extracts. The in vitro antioxidant activity was investigated with DPPH radical scavenging assay. The quantitative evaluation of DPPH scavenging activity showed that n-BuOH and EtOAc extracts are the most active extracts with a percentage of antiradical activity of 89,62% and 71,57% respectively.

Keywords: Launaea, phytochemical, South Algeria, Sahara, endemic specie

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Authors: Pakakrong Thongdeeying, Srisopa Ruangnoo, Arunporn Itharat

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The rhizomes of Smilax corbularia Kunth have long been used as common ingredients in anticancer preparations. Thus, the objective of this study is to investigate cytotoxicity of S. corbularia and its ingredients against cholangiocarcinoma cell line (KKU-M156) by SRB assay. Ethanolic and water extracts of S. corbularia rhizomes were obtained using the procedures followed by Thai traditional doctors. Bioassay guided isolation was used to isolate cytotoxic compounds. The results revealed that the ethanolic extract of S. corbularia exhibited activity against KKU-M156 cell line with an IC50 value of 84.53±1.62 µg/ml, but the water extract showed no cytotoxic activity. Three flavonoid compounds [astilbin (1), engeletin (2), and quercetin (3)] were isolated from the ethanolic extract. Compound 3 exhibited the strongest activity against KKU-M156 cell line (IC50 = 8.14 ± 1.15 µg/ml), but 1 and 2 showed no cytotoxic activity (IC50 > 100 µg/ml). In conclusion, quercetin showed the highest efficacy against cholangiocarcinoma. These results support the traditional use of this plant by Thai traditional doctors for cancer treatment.

Keywords: cholangiocarcinoma, cytotoxicity, flavonoid, Smilax corbularia

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Authors: Bensaid Fatiha

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The objective of our present work is the description of the antagonistic capacities of one strain of Pseudomonas fluorescens and the nonpathogenic fungic isolate Fusarium oxysporum against phytopathogenic agent Fusarium oxysporum F. Sp. lycopersici. This work has been achieved in two main parts: the first is interested on the in vitro antagonistic activities; the second was interested to study the soil receptiveness of fusarium wilt tomato. The use of strain of fluorescent Pseudomonas and a non-pathogenic strain of F. oxysporum in the different antagonism tests, has allowed assuring a certain bio-protection from the plants of tomatoes opposite to F. oxysporum F. Sp. lycopersici, agent of a wilt of tomato. These antagonistic have shown a substantial in vitro antagonistic activity on the three mediums (KB, PDA, KB+PDA) against F. oxysporum F. Sp. lycopersici, by inhibiting its growth mycelium with rate of inhibition going until 80 % with non-pathogen of Fusarium oxysporum and 60 % with strain of fluorescens Pseudomonas. Soil microbial balance, between the antagonistic population and that of pathogenic, can be modulated through microbiological variations or abiotic additives influencing directly or indirectly the metabolic behavior microbial. In this experiment, addition of glucose or EDTA, could increase or decrease the resistance of soil by activation of pathogenic or antagonists, as a result of modification and modulation in their metabolic activities.

Keywords: fluorescents, nonpathogenic, fusarium oxysporum, fusarium wilt, antagonism, biological control, soil receptivity

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Authors: Meksem Nabila, Bordjiba Ouahiba, Meraghni Messaouda, Meksem Amara Leila, Djebar Mohhamed Reda

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The problems of protection of the cultures are being more and more important that they interest great number of farmers and scientists because of the excessive use of the organic phytosanitary products of synthesis that causes fatal damages on the environment. To reduce the inconveniences produced by these pesticides, the use of "biopesticides" originated from plants could be an alternative. The aim of this work is the valuation of a botanical species: Eucalyptus camaldulensis from Northeastern Algeria which extracts are supposed to have an antimicrobial activity, similar to pesticides. The extraction of secondary metabolites from the leaves of E. camaldulensis was realized using methanol and water, and measurements of total polyphenols were made by spectrometric method. Determination of the antimicrobial activity of the extracts at issue was realized in vitro on phyto-pathogenic fungal and bacterial stumps. Tests of comparison were included in the essays by using the chemical pesticidal products of synthesis. The obtained results show that the plant contains polyphenols with an efficiency mattering of the order of 22 %. These polyphenols have a strong fungicidal and bactericidal pesticidal activity against various microbial stumps and the values of the zones of inhibition are more important compared with that obtained in the presence of the chemicals of synthesis (fungicide).

Keywords: eucalyptus camaldulensis, biopesticide, polyphenols, antimicrobial activity

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Authors: Alemu Tadesse Feroche

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In this study, some 2, 3 distributed quinazoline -4 (3H) - one derivative were synthesized using a three-step synthetic route. They were obtained in a good yield (59.5-85%) by applying different chemical reactions like cyclization and condensation reactions. The chemical structure of the final compounds was also verified by spectroscopic methods (IR, ¹HNMR) and elemental microanalysis. The in vivo antimalarial activity of these compounds on P. berghei infected mice was found to be moderate to high at an oral dose of 0.04846 mmol/kg /day. This is equal to 25 mg/kg of chloroquine phosphate, which causes 100% inhibition of the parasite. It is worth mentioning that most active compounds (E) -3 Phenyl -2- [2- (pyridine -4- yl) vinyl] -4 (3H) -quinazolinone IVa (64.02%, (E)-2-[2-(4 - Hydroxy-3 - methoxystyryl) - vinyl) -3 - phenyl -4 (3H ) - quinazolinone IVc (77.25%) and (E)-2 –[2 –(Pyridin -4-yl) –vinyl] -3 phenenylamine -4(3H) quinazolinone IVe (73.54%) showed a dose-dependent increase in present suppression in antimalarial activities. Furthermore, the synthesized compounds were screened for their in vitro antileishmanial activity against L. aethiopica isolate (CL/039/09). All tested compounds (IVa (0.03766 ug/ml), IVb (0.00538 ug/ml, IVc (0.00412 ug/ml, IVd (0.00110 ug/ml), IVe (0.03017 ug/ml) and IVf (0.03894 ug/ml)) showed excellent potency that is much better than amphotericin B (IC50 = 0,04359 ug/ml). The results of acute toxicity indicated that all test compounds (IVa –IVf) proved to be nontoxic and well tolerated by the experimental animals up to 300 mg/kg in oral and 140 mg/kg in parental studies.

Keywords: 4(3H)-quinazolinone, in vivo antimalarial activity, in vitro antileishmanial activity, acute toxicity

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Authors: Ririn Lestari Sri Rahayu, Mustofa Ahda

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The use of synthetic antioxidants often causes a negative effect on health and increases the incidence of carcinogenesis. Development of the natural antioxidants should be investigated. However, natural antioxidants have a low toxicity and are safe for human consumption. Ethanol extract of mangosteen rind (Garcinia mangostana) contains natural antioxidant compounds that have various pharmacological activities. Antioxidants from the ethanol extract of mangosteen rind have free radicals scavenging activities. The scavenging activity of ethanol extract of mangosteen rind was determined by DPPH method. The phenolic compound from the ethanol extract of mangosteen rind is determined with Folin-Ciocalteu method. The results showed that the absolute ethanol extract of mangosteen rind has IC₅₀ of 40.072 ug/mL. The correlation of total phenols content with free radical scavenging activity has an equation y: 5.207x + 205.51 and determination value (R²) of 0.9329. Total phenols content from the ethanol extract of mangosteen rind has a good correlation with free radicals scavenging activity of DPPH.

Keywords: Antioxidant, Garcinia mangostana, Inhibition concentration 50%, Phenolic.

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Authors: Hyun Joon Chang, Jae In Kim, Sungsoo Na

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Kinesin-1 (hereafter called kinesin) is a molecular motor protein that moves cargos toward the end of microtubules using the energy of adenosine triphosphate (ATP) hydrolysis. When kinesin is inactive, its tail autoinhibits the motor chain in order to prevent from reacting with the ATP by cross-linking of the tail domain to the motor domains at two positions. However, the morphological study of kinesin during autoinhibition is yet remained obscured. In this study, we report the effect of the binding site of the tail domain using the normal mode analysis of the elastic network model on kinesin in the tail-free form and tail-bind form. Considering the relationship between the connectivity of conventional network model with respect to the cutoff length and the functionality of the binding site of the tail, we revaluated the network model to observe the key role of the tail domain in its structural aspect. Contingent on the existence of the tail domain, the results suggest the morphological stability of the motor domain. Furthermore, employing the results from normal mode analysis, we have determined the strain energy of the neck linker, an essential portion of the motor domain for ATP hydrolysis. The results of the neck linker also converge to the same indication, i.e. the morphological analysis of the motor domain.

Keywords: elastic network model, Kinesin-1, autoinhibition

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Authors: Manal Kamel, Sara Maher

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Development of sensitive non-invasive tests for detection of SARS-CoV-2 antigens is imperative to manage the extent of infection throughout the population, yet, it is still challenging. Here, we designed and optimized a sandwich enzyme-linked immunosorbent assay (ELISA) for SARS-CoV-2 S1 antigen detection in saliva. Both saliva samples and nasopharyngeal swapswere collected from 170 PCR-confirmed positive and negative cases. Gold nanoparticles (AuNPs) were conjugated with S1protein receptor binding domain (RBD) nanobodies. Recombinant S1 monoclonal antibodies (S1mAb) as primery antibody and gold conjugated nanobodies as secondary antibody were employed in sandwich ELISA. Our developed system were optimized to achieve 87.5 % sensitivity and 100% specificity for saliva samples compared to 89 % and 100% for nasopharyngeal swaps, respectively. This means that saliva could be a suitable replacement for nasopharyngeal swaps No cross reaction was detected with other corona virus antigens. These results revealed that our developed ELISAcould be establishedas a new, reliable, sensitive, and non-invasive test for diagnosis of SARS-CoV-2 infection, using the easily collected saliva samples.

Keywords: COVID 19, diagnosis, ELISA, nanobodies

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Authors: Byeal-I Han, Michael Lee

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Ageing is associated with an increased risk of diseases such as cancer, and neurodegenerative disorders. Increased autophagy delays ageing and extends longevity. In this study, we investigated the role of autophagy in longevity using human foreskin fibroblast HS68 cells, in which a senescence-like growth arrest can be induced. In particular, cellular senescence is manifested by the irreversible cell cycle arrest, and may contribute to the ageing of organisms. The senescence state was measured with staining for senescence-associated β-galactosidase (SA-β-gal) activity that represents a sensitive and reliable marker to quantify senescent cells. We detected a significantly increased percentage (%) of SA-β-gal positive cells in HS68 cultures at passage 40 (63%) when compared with younger ones at passage 15 (0.5%). As expected, HS68 cells at passage 40 exhibited much lower proliferation rate than cells at passage 15. The basal levels of LC3 were measured by immunoblotting showing a comparison of LC3-I and LC3-II levels at 3 age-points in serially passaged HS68 cells. LC3-II/LC3-I ratio at different passage levels relative to β-actin levels of each band confirmed that cells at passage 34 showed lower conversion of non-autophagic LC3-I to autophagic LC3-II than the cells at passage 16. Furthermore, Cyto-ID autophagy assay also revealed that late passage cells showed lower autophagy than the early passage cells. Together, our findings suggest that senescence induction might be associated with decreased autophagy.

Keywords: ageing, autophagy, senescence, HS68

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Authors: M. Dezyani, R. Ezzati belvirdi, M. Shakerian, H. Mirzaei

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The effect of Microfiltration (MF) on proteolysis, hardness, and flavor of Feta cheese during 6 mo of aging was determined. Raw skim milk was microfiltered two-fold in two cheese making trials. In trial 1, four vats of cheese were made in 1 d using unconcentrated milk (1X), 1.26X, 1.51X, and 1.82X Concentration Factors (CF). Casein-(CN)-to-fat ratio was constant among treatments. Proteolysis during cheese aging decreased with increasing CF due to either limitation of substrate availability for chymosin due to low moisture in the nonfat substance (MNFS), inhibition of chymosin activity by high molecular weight milk serum proteins, such as α2-macroglobulin, retained in the cheese or low residual chymosin in the cheese. Hardness of fresh cheese increased, and cheese flavor intensity decreased with increasing CF. In trial 2, the 1X and 1.8X CF were compared directly. Changes made in the cheese making procedure for the 1.8X CF (more chymosin and less cooking) increased the MNFS and made proteolysis during aging more comparable for the 1X and 1.8X cheeses. The significant difference in cheese hardness due to CF in trial 1 was eliminated in trial 2. In a triangle test, panelists could not differentiate between the 1X and 1.8X cheeses. Therefore, increasing chymosin and making the composition of the two cheeses more similar allowed production of aged Fetta cheese from milk concentrated up to 1.8X by MF that was not perceived as different from aged feta cheese produced without MF.

Keywords: feta cheese, microfiltration, concentration factor, proteolysis

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Authors: Abdelkader Basli, Jean-Claude Delaunay, Eric Pedrot, Jean-Michel Mérillon, Jean-Pierre Monti, Khodir Madani, Mohamed Chibane, Tristan Richard

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The antioxidant and antimicrobial activities of Origanum glandulosum collected in Algeria have been studied. Extract was prepared from aerial part of endemic Algerian oregano. The produced extract has been characterized in terms of total phenols (using Folin method), total flavonoid, antioxidant activities (using the DPPH radical scavenging method and ORAC assay) and microbial activity against four bacteria: Streptococcus aureus, Bacillus subtilis, Escherichia coli, Klebsiella pneumoniae one yeast: Candida albicans and one fungi: Aspergillus niger. The results pointed the antioxidant activities of the extract of O. glandulosum and antimicrobial activities against all bacteria and C. Candida, but no effect on A. niger. High performance liquid chromatography combined with mass spectrometry (LC-MS) and nuclear magnetic resonance (LC-NMR) were used to separate and identify the major compounds present in the oregano extract. Rosmarinic acid, globoidnan A and B, lithospermic acid B and three flavonoids were identified.

Keywords: origanum glandulosum, antioxidant, microbial activity, polyphenol, LC-MS, LC-NMR

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Authors: Sahira Nsayef Muslim, Israa M. S. Al-Kadmy, Nadheema Hammood Hussein, Alaa Naseer Mohammed Ali, Buthainah Mohammed Taha, Rayim Sabah Abbood, Sarah Naji Aziz

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A novel strain of Bacillus licheniformis isolated from spoilt cucumber and pepper samples have the ability to produce the chitosanase enzyme when grown on chitosan substrate. Chitosanase was purified to homogeneity with a recovery yield of 35.71% and 5.5 fold of purification by using ammonium sulfate at 45% saturation followed by ion exchange chromatography on DEAE-cellulose column and gel filtration chromatography on Sephadex G-100 column. The purified chitosanase inhibited the biofilm formation ability for all Gram-negative and Gram-positive biofilm-forming bacteria (biofilm producers) after using Congo Red agar and Microtiter plates methods. Highly antibiofilm of chitosanase recorded against Pseudomonas aeruginosa followed by Klebsiella pneumoniae with reduction of biofilm formation ratio to 22 and 29%, respectively compared with (100)% of control. Thus, chitosanase has promising benefit as antibiofilm agent against biofilm forming pathogenic bacteria and has promising application as alternative antibiofilm agents to combat the growing number of multidrug-resistant pathogen-associated infections, especially in situation where biofilms are involved.

Keywords: chitosanase, Bacillus licheniformis, vegetables, biofilm

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Authors: G. V. Novikov, V. S. Sivozhelezov, S. S. Kolesnikov, K. V. Shaitan

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The study reports about the influence of binding of orthosteric ligands as well as point mutations on the conformational dynamics of β-2-adrenoreceptor. Using molecular dynamics simulation we found that there was a little fraction of active states of the receptor in its apo (ligand free) ensemble corresponded to its constitutive activity. Analysis of MD trajectories indicated that such spontaneous activation of the receptor is accompanied by the motion in intracellular part of its alpha-helices. Thus receptor’s constitutive activity directly results from its conformational dynamics. On the other hand the binding of a full agonist resulted in a significant shift of the initial equilibrium towards its active state. Finally, the binding of the inverse agonist stabilized the receptor in its inactive state. It is likely that the binding of inverse agonists might be a universal way of constitutive activity inhibition in vivo. Our results indicate that ligand binding redistribute pre-existing conformational degrees of freedom (in accordance to the Monod-Wyman-Changeux-Model) of the receptor rather than cause induced fit in it. Therefore, the ensemble of biologically relevant receptor conformations is encoded in its spatial structure, and individual conformations from that ensemble might be used by the cell in conformity with the physiological behaviour.

Keywords: seven-transmembrane receptors, constitutive activity, activation, x-ray crystallography, principal component analysis, molecular dynamics simulation

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Authors: Nootchanat Mairuae, Walaiporn Tongjaroenbuangam, Chalisa Louicharoen Cheepsunthorn, Poonlarp Cheepsunthorn

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The aim of this study was to investigate the anti-oxidative effect, the anti-inflammatory effects, and the molecular mechanisms of okra (Abelmoschus esculentus Linn.) on lipopolysaccharide (LPS)-stimulated BV2 microglial cells. The BV2 cells were treated with LPS in the presence or absence of okra. Reactive oxygen species (ROS) and nitric oxide (NO) production were measured using the ROS detection reagent DCF-DA and the Griess reaction, respectively. The phosphorylation levels of nuclear factor-kappa B (NF-kB) p65 was detected by Western blot assay. Treatment of BV2 microglia cells with okra was found to significantly suppress the LPS-induced inflammatory mediator NO as well as ROS compared to untreated cells. The levels of LPS-induced NF-kB p65 phosphorylation were significantly decreased following okra treatment too. These results show that okra exerts anti-oxidative and anti-inflammatory effects in LPS-stimulated BV2 microglial cells by suppressing the NF-κB pathway. This suggests okra might be a valuable agent for treatment of anti-neuroinflammatory diseases mediated by microglial cells.

Keywords: Abelmoschus esculentus Linn, microglia, neuroinflammation, reactive oxygen spicy

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Authors: Rafat M. Mohareb

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Several D-ring alkylated estrone analogues display exceptionally high affinity for estrogen receptors. In particular, compounds in which an E-ring is formed are known to be involved in the inhibition of steroidogenic enzymes. Such compounds also have an effect on steroid dehydrogenase activity and the ability to inhibit the detrimental action of the steroid sulfatase enzyme. Generally, E-ring extended steroids have been accessed by modification of the C17-ketone in the D-ring by either arylimine or oximino formation, addition of a carbon nucleophile or hydrazone formation. Other approaches have included ketone reduction, silyl enol ether formation or ring-closing metathesis (giving five- or six-membered E-rings). Chemical modification of the steroid D-ring provides a way to alter the functional groups, sizes and stereochemistry of the D-ring, and numerous structure-activity relationships have been established by such synthetic alterations. Steroids bearing heterocycles fused to the D-ring of the steroid nucleus have been of pharmaceutical interest. In the present paper, we report on the efficient synthesis of estrone possessing pyran, pyrimidine and thiazole ring systems. This study focused on the synthesis and biochemical evaluation of newly synthesized heterocyclic compounds which were then subjected through inhibitory evaluations towards human cancer and normal cell lines.

Keywords: estrone, heterocyclization, cytotoxicity, biomedicine

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Authors: Sahriye Sonmez, Sedat Citak

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The objective of this study was to evaluate the effects of different organic manures on antioxidant activity, vitamin C and nitrate concentrations of broccoli (Brassica oleracea L. var italica) plants. For this purpose, broccoli plants were grown on open field conditions in 2 successive years (2011-2013) including 4 different seasons [(Spring 1 (March-June, 2011), Autumn 1 (September 2011-January 2012), Spring 2 (March-June, 2012), Autumn 2 (September 2012-January 2013)]. Organic manures (Farm manure (FM), vermicompost (VC) and leonardite (L) and its mixture (50 % FM+50% L, 50 % VC+50% FM, 50% L+50% VC and 33% FM+33% VC+33% L), one chemical fertilizer and one control, collectively 9 applications was investigated. The results indicated that the vitamin C concentrations of broccoli plants ranged from 31.4-55.8 mg/100 g, 43-631 mg/kg in nitrate concentrations and 11.0-56.7 mg/ml as IC50 inhibition values in antioxidant activities of broccoli plants. Also, it was determined that the effective applications were at the 50 % VC+50% FM for vitamin C concentrations, at the chemical fertilizer for nitrate concentrations and at the 100 % FM for antioxidant activities.

Keywords: broccoli, chemical fertilizer, farm manure, leonardite, vermicompost

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Authors: Lourena M. Veríssimo, Lucas A. Machado, Renata Rutckeviski, Francisco H. Xavier Júnior, Éverton N. Alencar, Andreza R. V. Morais, Teresa R. F. Dantas, Christian M. Oliveira, Arnóbio A. Silva Júnior, Eryvaldo S. T. Egito

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This study aimed to obtain emulsion systems based on bullfrog oil (BO). The BO was extracted at 80ºC and analyzed by Gas Chromatography-Mass Spectrometry (GC/MS). The critical Hydrophilic-Lipophilic Balance (HLBc) Assay of the BO was performed through BO, Tween® 20, Span® 80 and deionized water mixtures using an Ultra-Turrax® and determined using dynamic light scattering, pH, electrical conductivity and creaming rate. Then, a pseudoternary phase diagram (PPD) was constructed by water titration. The GC/MS analysis of BO suggested Methyl Oleate (9.26%) as major compound. The HLBc was 12.1, wherein the correspondent emulsion showed a pH of 4.83±1.29, electrical conductivity of 103.65 µS, creaming rate of 2.51±0.54%, droplet size of 207.07±8.31 nm and polydispersity index of 0.212±0.005. The PPD showed different formulations characterized as O/W emulsions. Thus, the PPD proved to be a useful tool to produce BO emulsions, in which their constituents may vary within the range of the desired system.

Keywords: bullfrog (Rana catesbeiana Shaw) oil, emulsion production, hydrophilic-lipophilic balance, gas chromatography/mass spectrometry analysis

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Authors: Jau-Shya Lee, Rossita Shapawi, Vin Cent Pua

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Tilapia is a popular cultured fresh-water fish in Malaysia. The highly perishable nature of the fish and increasing demand for high-quality ready-to-cook fish has intensified the search for better fish preservation method. Chitosan edible coating has been evident to extend the shelf life of fish fillet. This work was attempted to explore the potential of ascorbic acid in enhancing the shelf life extension ability of chitosan coated Tilapia fillet under refrigeration condition (4 ± 1oC). A 3 2 Factorial Design which comprising of three concentrations of chitosan (1, 1.5 and 2%) and two concentrations of ascorbic acids (2.5 and 5%) was used. The fish fillets were analyzed for total viable count, thiobarbituric acid (TBA) value, pH, aw and colour changes at 3-day interval over 15-day storage. The shelf life of chitosan coated (1.5% and 2%) fillet was increased to 15 days as compared to uncoated fish fillet which can only last for nine days. The inhibition of microbial growth of fish fillet was enhanced with the addition of 5% of ascorbic acids in 2% of chitosan. The TBA value, pH and aw for chitosan coated samples were found lower than that of uncoated sample (p<0.05). The colour stability of the fish fillet was also improved by the composite coating. Overall, 2% of chitosan and 5% of ascorbic acid formed the most effective coating to enhance the quality and to lengthen the shelf life of refrigerated Tilapia fillet.

Keywords: ascorbic acid, chitosan, edible coating, fish fillet

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Authors: Chih-Hung Liang, Jung-Jung Chen, Shen-Shih Chiang

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Obesity is defined as abnormal or excessive fat accumulation that presents a risk to health, which are major risk factors for a number of chronic diseases, including diabetes, cardiovascular diseases and cancer. Cordyceps militaris (CM) is a well-known traditional medicine in Asian countries and a rich source of biologically active components. Black rice (Oryza sativa L.) is a special cultivar of rice that contains rich anthocyanins and regarded as a health-promoting food in China and other Eastern. The aim of this study was to investigate the anti-obesity effect of Cordyceps militaris fermented black rice (CB) on HFD-induced BALB/c mice model. The results indicated that administration of low and high dosage of CB powder significantly reduced the body weights (7.38% and 7.78%), body fat ratio (2.37% and 2.78%), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels compared to the HF group (p<0.05). Histopathological analysis showed that the score of fatty liver in HF group (5.0) was significantly higher than CB groups (2.1 and 3.6) (p<0.05). In conclusion, Cordyceps militaris fermented black rice can reduce the body weight via inhibition of the fat accumulation in liver and body and possess the anti-obesity potency.

Keywords: Cordyceps militaris, black rice, obesity, HFD-induced mice

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Authors: Salahaedin Waiezi, Nik Ahmad Nizam Nik Malek, Hassan Abdelmagid Elzamzami, Shahrulnizahana Mohammad Din

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A green and safe approach to the synthesis of silver nanoparticles (AgNP) can be performed using plant leaf extract as the reducing agent. Hence, this paper reports the biosynthesis of AgNP using Clinacanthus nutans plant extract. C. nutans is known as belalai gajah in Malaysia and is widely used as a medicinal herb locally. The biosynthesized AgNP, using C. nutans aqueous extract at pH 10, with the reaction temperature of 70°C and 48 h reaction time, was characterized by UV-Vis spectroscopy, X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM), energy dispersive X-ray (EDX), and transmission electron microscope (TEM). A peak appeared in the UV-Vis spectra at around 400 nm, while XRD confirmed the crystal structure of AgNP, with the average size between 20 to 30 nm, as shown in FESEM and TEM. The antibacterial activity of the biosynthesized AgNP, which was performed using the disc diffusion technique (DDT) indicated effective inhibition against Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus aureus. In contrast, minimal antibacterial activity was detected against Enterococcus faecalis and methicillin-resistant Staphylococcus aureus (MRSA). In general, AgNP produced using C. nutans leaf extract possesses potential antibacterial activity.

Keywords: silver nanoparticles, Clinacanthus nutans, antibacterial agent, biosynthesis

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Authors: Chao Wu

Abstract:

Thioredoxin reductase 2 is a mitochondrial enzyme that belongs to the cellular defense against oxidative stress. We deleted mitochondrial Txnrd2 in a KrasG12D-driven pancreatic tumor model. Despite an initial increase in precursor lesions, tumor incidence decreased significantly. We isolated cancer cell lines from these genetically engineered mice and observed an impaired proliferation and colony formation. Reactive Oxygen Species, as determined by DCF fluorescence, were increased. We detected a higher mitochondrial copy number in Txnrd2-deficient cells (KTP). However, measurement of mitochondrial bioenergetics showed no impairment of mitochondrial function and comparable O₂-consumption and extracellular acidification rates. In addition, the mitochondrial complex composition was affected in Txnrd2 deleted cell lines. To gain better insight into the role of Txnrd2, we deleted Txnrd2 in clones from parental KrasG12D cell lines using Crispr/Cas9 technology. The deletion was confirmed by western blot and activity assay. Interestingly, and in line with previous RNA expression analysis, we saw changes in EMT markers in Txnrd2 deleted cell lines and control cell lines. This might help us explain the reduced tumor incidence in KrasG12D; Txnrd2∆panc mice.

Keywords: PDAC, TXNRD2, epithelial-to-mesenchymal-transition, ROS

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Authors: V. Yurina, H. Sujuti, E. Rahmani, A. R. Nopitasari

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Breast cancer has the highest prevalence cancer in women. Moringa leaves (M. oleifera) contain quercetin, kaempferol, and benzyl isothiocyanate which can enhance induction of apoptosis. This research aimed to study the role of the leaf extract of Moringa to increase apoptosis in breast cancer cell line, MCF-7 cells. This research used in vitro experimental, post-test only, control group design on breast cancer cells MCF-7 in vitro. Moringa leaves were extracted by maceration method with ethanol 70%. Cells were treated with drumstick leaves extract on 1100, 2200, and 4400 μg/ml for Hsp27 and caspase-9 expression (immunocytochemistry) and apoptosis (TUNEL assay) test. The results of this study found that the IC50 2200 µg/ml. Moringa leaves extract can significantly increase the expression of caspase-9 (p<0.05) and decreased Hsp 27 expression (p<0.05). Moreover it can increase apoptosis (p<0.05) significantly in MCF-7 cells. The conclusion of this study is Moringa leaves extract is able to increase the expression of caspase-9, decrease Hsp27 expression and increase apoptosis in breast cancer cell-line MCF-7.

Keywords: apoptosis, breast cancer, caspase-9, Hsp27, Moringa oleifera

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Authors: B. Loukil, L. Mallem, M. S. Boulakoud

Abstract:

Inorganic fertilizers consist mainly of salts of ammonium nitrate, phosphate and potassium, the combination of primary nutrients NPK including secondary and micro nutrients are essential for plant growth, used for intensive agriculture, ranching, and horticultural crops, to increase soil fertility and ensure sustainable crop production. The manufacture of fertilizers is generally at a high temperature and high pressure, in the presence of several highly hazardous chemicals, dust and gases. These products are absorbed high in the airway, increasing the airway resistance thereby adversely affecting the pulmonary functions of workers. A study was conducted on 34 employees, especially exposed to nitrate derivatives. A questionnaire was prepared and distributed to all employees in the unit. The workers were divided into two groups according to age. Several hormonal parameters Assay were measured. The results of the questionnaire have detected a fertility problem, Concerning the hormones a significant reduction in the concentration of testosterone in both groups and LH in the group aged 30 to 40 year were noted compared to the control. However, an increase in the concentration of prolactin in both groups compared to the control. There was a significant decrease in FSH in the group aged 30 to 40 always in compared with the control group.

Keywords: fertilizers, healthy worker, risk, fertility

Procedia PDF Downloads 392

Authors: Isil Gazioglu, Abdulselam Ertas

Abstract:

Aflatoxins (AFs) are secondary toxic metabolites of Aspergillus flavus and A. parasiticus. AFs can be absorbed through the skin. Potent carcinogens like AFs should be completely absent from cosmetics, this can be achieved by careful quality control of the raw plant materials. Regulatory limits for aflatoxins have been established in many countries, and reliable testing methodology is needed to implement and enforce the regulatory limits. In this study, ten medicinal plant samples (Bundelia tournefortti, Capsella bursa-pastoris, Carduus tenuiflorus, Cardaria draba, Malva neglecta, Malvella sharardiana, Melissa officinalis, Sideritis libanotica, Stakys thirkei, Thymus nummularius) were investigated for aflatoxin (AF) contaminations by employing an HPLC assay for the determination of AFB1, B2, G1 and G2. The samples were extracted with 70% (v/v) methanol in water before further cleaned up with an immunoaffinity column and followed by the detection of AFs by using an electrochemically post-column derivatization with Kobra-Cell and fluorescence detector. The extraction procedure was optimized in order to obtain the best recovery. The method was successfully carried out with all medicinal plant samples. The results revealed that five (50%) of samples were contaminated with AFs. The association between particular samples and the AF contaminated could not be determined due to the low frequency of positive samples.

Keywords: aflatoxin B1, HPLC-FLD, KOBRA-Cell, mycotoxin

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Authors: Harukuni Tokuda, Xu FengHao, Nobutaka Suzuki

Abstract:

As part of an ongoing our projects to investigate the anti-tumor promoring properties (chemopreventive potency) of Gromwell seed, dry powder materials and its active compounds were carried out through useful test systems. Gromwell seed (Coix lachryma-jobi seed) (GS) is a grass crop that has long been used and played a role in traditional medicine as a nourishing food, and for the treatment of various aliments, paticularly cancer. The application of a new screening procedure which utilizes the synergistic effect of short-chain fatty acids and phorbol esters in enable rapid and easy detection of naturally occurring substances(anti-tumor promoters chemo-preventive agents) with inhibition of Epstein-Barr virus(EBV) activation, using human lymphblastoid cells. In addition, we have now extended these investigations to a new tumorigenesis model in which we initiated the tumors with DMBA intiation and promoted with 1.7 nmol of TPA in two-stage mouse skin test and other models. these results provide a basis for further development of these botanical supplements for human cancer chemoprevention and observations seem that this materials more extensively as one of the trials for the purpose of complementary and alternative medicine.

Keywords: chemoprevention, medicinal plant, mouse, carcinogenesis systems

Procedia PDF Downloads 476

Authors: Duygu Yüksel Deniz, Memet Vezir Kahraman, Serap Erdem Kuruca, Mediha Süleymanoğlu

Abstract:

Our first aim was to synthesize Hydroxy Apatite (HAP) and then modify its surface by adding 4-Vinylbenzene boronic acid (4-VBBA). The characterization was done by FT-IR. By adding Polyvinyl alcohol (PVA) to 4- VBBA-HAP, we obtained a suitable electrospinning solution. PVA solution which was also modified by using alkoxy silanes, in order to prevent the scaffolds from being damaged by aqueous cell medium, was added. Keratin was dissolved and then added into the electrospinning solution. Keratin containing 4-VBBA- HAP/PVA composite was used to fabricate nanofiber scaffolds with the simultaneous UV-reactive electrospinning technique. The structural characterization was done by FT-IR. Thermal gravimetric analysis was also performed by using TGA. The morphological characterization was determined by SEM analyses. Our second aim was to create a scaffold where cells could grow. With this purpose, suitable nanofibers were choosen according to their SEM analysis. Keratin containing nanofibers were seeded with 3T3, ECV and SAOS cells and their cytotoxicity and cell proliferation were investigated by using MTT assay. After cell culturing process morphological characterization was determined by SEM analyses. These scaffolds were designed to be nontoxic biomaterials. Here, a comparision was made between keratin containing 3T3, ECV and SAOS seeded nanofiber scaffolds and the results were presented and discussed.

Keywords: cell culture, keratin, nanofibers, UV-reactive electrospinning

Procedia PDF Downloads 445