Search results for: assay parameters

Authors: A. A. Okafor, C. H. Achebe, J. L. Chukwuneke, C. G. Ozoegwu

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The performance of an engine whose basic design parameters are known can be predicted with the assistance of simulation programs into the less time, cost and near value of actual. This paper presents a comprehensive mathematical model of the performance parameters of four stroke spark ignition engine. The essence of this research work is to develop a mathematical model for the analysis of engine performance parameters of four stroke spark ignition engine before embarking on full scale construction, this will ensure that only optimal parameters are in the design and development of an engine and also allow to check and develop the design of the engine and it’s operation alternatives in an inexpensive way and less time, instead of using experimental method which requires costly research test beds. To achieve this, equations were derived which describe the performance parameters (sfc, thermal efficiency, mep and A/F). The equations were used to simulate and optimize the engine performance of the model for various engine speeds. The optimal values obtained for the developed bivariate mathematical models are: sfc is 0.2833kg/kwh, efficiency is 28.77% and a/f is 20.75.

Keywords: bivariate models, engine performance, injector engine, optimization, performance parameters, simulation, spark ignition

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Authors: Carlos Caro, Ernest Blade, Nestor Rojas

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This study determined the relationship between basic geo-technical parameters and parameters of the hydro logical model Thomas for water balance of rural watersheds, as a methodological calibration application, applicable in distributed models as IBER model, which represents a distributed system simulation models for unsteady flow numerical free surface. There was an exploration in 25 points (on 15 sub) basin of Rio Piedras (Boy.) obtaining soil samples, to which geo-technical characterization was performed by laboratory tests. Thomas model has a physical characterization of the input area by only four parameters (a, b, c, d). Achieve measurable relationship between geo technical parameters and 4 values of hydro logical parameters helps to determine subsurface, underground and surface flow more agile manner. It is intended in this way to reach some solutions regarding limits initial model parameters on the basis of Thomas geo-technical characterization. In hydro geological models of rural watersheds, calibration is an important process in the characterization of the study area. This step can require a significant computational cost and time, especially if the initial values or parameters before calibration are outside of the geo-technical reality. A better approach in these initial values means optimization of these process through a geo-technical materials area, where is obtained an important approach to the study as in the starting range of variation for the calibration parameters.

Keywords: distributed hydrology, hydrological and geotechnical characterization, Iber model

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Authors: Mahnoor Khan, Bashir Ahmad, Khizar Hayat, Saad Ahmad Khan, Laiba Ahmad, Shumaila Bashir, Abid Ali Khan

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The objective of this study was to synthesize the smallest possible size of ZnO NPs using a modified wet chemical synthesis method and to prepare core shell using polyethylene glycol (PEG) as shell material. Advanced and sophisticated techniques were used to confirm the synthesis, size, and shape of these NPs. Rounded, clustered NPs of size 5.343 nm were formed. Both the plain and core shell NPs were tested against MDR bacteria (E. cloacae, E. amnigenus, Shigella, S. odorifacae, Citrobacter, and E. coli). Both of the NPs showed excellent antibacterial properties, whereas E. cloacae showed maximum zone of inhibition of 16 mm, 27 mm, and 32 mm for 500 μg/ml, 1000 μg/ml, and 1500 μg/ml, respectively for plain ZnO NPs and 18 mm, 28 mm and 35 mm for 500 μg/ml, 1000 μg/ml and 1500 μg/ml for core shell NPs. These NPs were also biocompatible on human red blood cells showing little hemolysis of only 4% for 70 μg/ml for plain NPs and 1.5% for 70 μg/ml for core shell NPs. Core shell NPs were highly biocompatible because of the PEG. Their therapeutic effect as photosensitizers in photodynamic therapy (PDT) for cancer treatment was also monitored. The cytotoxicity of ZnO and PEG-ZnO was evaluated using MTT assay. Our results demonstrated that these NPs could generate ROS inside tumor cells after irradiation which in turn initiates an apoptotic pathway leading to cell death hence proving to be an effective candidate for PDT.

Keywords: ZnO, hemolysis, cytotoxiciy assay, photodynamic therapy, antibacterial

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Authors: Mohamed Noureldin, Jinkoo Kim

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The sensitivity of the seismic response parameters to the uncertain modeling variables of pile-founded fixed steel jacket platforms are investigated using tornado diagram, first-order second-moment, and static pushover analysis techniques. The effects of both aleatory and epistemic uncertainty on seismic response parameters have been investigated for an existing offshore platform. The sources of uncertainty considered in the present study are categorized into three different categories: the uncertainties associated with the soil-pile modeling parameters in clay soil, the platform jacket structure modeling parameters, and the uncertainties related to ground motion excitations. It has been found that the variability in parameters such as yield strength or pile bearing capacity has almost no effect on the seismic response parameters considered, whereas the global structural response is highly affected by the ground motion uncertainty. Also, some uncertainty in soil-pile property such as soil-pile friction capacity has a significant impact on the response parameters and should be carefully modeled. Based on the results, it is highlighted that which uncertain parameters should be considered carefully and which can be assumed with reasonable engineering judgment during the early structural design stage of fixed steel jacket platforms.

Keywords: fixed jacket offshore platform, pile-soil structure interaction, sensitivity analysis

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Authors: Hanan Khojah, RuAngelie Edrada-Ebel

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Neurodegenerative disease including Alzheimer’s disease is a major cause of long-term disability. Oxidative stress is frequently implicated as one of the key contributing factors to neurodegenerative diseases. Protection against neuronal damage remains a great challenge for researchers. Ficus carica (commonly known as fig) is a species of great antioxidant nutritional value comprising a protective mechanism against innumerable health disorders related to oxidative stress as well as Alzheimer’s disease. The purpose of this work was to characterize the non-polar active metabolites in Ficus carica endocarp, mesocarp, and exocarp. Crude extracts were prepared using several extraction solvents, which included 1:1 water: ethylacetate, acetone and methanol. The dried extracts were then solvent partitioned between equivalent amounts of water and ethylacetate. Purification and fractionation were accomplished by high-throughput chromatography. The isolated metabolites were tested on their effect on human neuroblastoma cell line by cell viability test and cell cytotoxicity assay with acrolein. Molecular weights of the active metabolites were determined via LC–HRESIMS and GC-EIMS. Metabolomic profiling was performed to identify the active metabolites by using differential expression analysis software (Mzmine) and SIMCA for multivariate analysis. Structural elucidation and identification of the interested active metabolites were studied by 1-D and 2-D NMR. Significant differences in bioactivity against a concentration-dependent assay on acrolein radicals were observed between the three fruit parts. However, metabolites obtained from mesocarp and the endocarp demonstrated bioactivity to scavenge ROS radical. NMR profiling demonstrated that aliphatic compounds such as γ-sitosterol tend to induce neuronal bioactivity and exhibited bioactivity on the cell viability assay. γ-Sitosterol was found in higher concentrations in the mesocarp and was considered as one of the major phytosterol in Ficus carica.

Keywords: alzheimer, Ficus carica, γ-Sitosterol, metabolomics

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Authors: Donna M. Morrison, Lambert Chester, Coretta A. N. Samuels, David R. Ledoux

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A survey was conducted in the five rice-growing regions in Guyana to determine the presence of aflatoxins in multiple fractions of rice in June/October 2015 growing season. The fractions were paddy, steamed paddy, cargo rice, white rice and parboiled rice. Samples were analyzed by High Performance Liquid Chromatography. A subset of the samples was further analyzed by enzyme-linked immunosorbent assay (ELISA) for concurrence. All analyses were conducted at the University of Missouri, USA. Of the 186 samples tested, 16 had aflatoxin concentrations greater than 20 ppb the recommended limit for aflatoxins in food according to the United States Food and Drug Administration. An additional three samples had aflatoxin B₁ concentrations greater than the European Union Commission maximum levels for aflatoxin B₁ in rice at 5 µg/kg and total aflatoxins (B₁, B₂, G₁ and G₂) at 10 µg/kg. The survey indicates that there is no widespread aflatoxin problem in rice in Guyana. The incidence of aflatoxins appears to be localized.

Keywords: aflatoxin, enzyme-linked immunosorbent assay (ELISA), high-performance liquid chromatography (HPLC), rice fractions

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Authors: Melanie Ostermann, Eivind Birkeland, Ying Xue, Alexander Sauter, Mihaela R. Cimpan

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Background: New reliable and robust techniques to assess biological effects of nanomaterials (NMs) in vitro are needed to speed up safety analysis and to identify key physicochemical parameters of NMs, which are responsible for their acute cytotoxicity. The central aim of this study was to validate and evaluate the applicability and reliability of an impedance-based flow cytometry (IFC) technique for the high-throughput screening of NMs. Methods: Eight inorganic NMs from the European Commission Joint Research Centre Repository were used: NM-302 and NM-300k (Ag: 200 nm rods and 16.7 nm spheres, respectively), NM-200 and NM- 203 (SiO₂: 18.3 nm and 24.7 nm amorphous, respectively), NM-100 and NM-101 (TiO₂: 100 nm and 6 nm anatase, respectively), and NM-110 and NM-111 (ZnO: 147 nm and 141 nm, respectively). The aim was to assess the biological effects of these materials on human monoblastoid (U937) cells. Dispersions of NMs were prepared as described in the NANOGENOTOX dispersion protocol and cells were exposed to NMs at relevant concentrations (2, 10, 20, 50, and 100 µg/mL) for 24 hrs. The change in electrical impedance was measured at 0.5, 2, 6, and 12 MHz using the IFC AmphaZ30 (Amphasys AG, Switzerland). A traditional toxicity assay, Trypan Blue Dye Exclusion assay, and dark-field microscopy were used to validate the IFC method. Results: Spherical Ag particles (NM-300K) showed the highest toxic effect on U937 cells followed by ZnO (NM-111 ≥ NM-110) particles. Silica particles were moderate to non-toxic at all used concentrations under these conditions. A higher toxic effect was seen with smaller sized TiO2 particles (NM-101) compared to their larger analogues (NM-100). No interferences between the IFC and the used NMs were seen. Uptake and internalization of NMs were observed after 24 hours exposure, confirming actual NM-cell interactions. Conclusion: Results collected with the IFC demonstrate the applicability of this method for rapid nanotoxicity assessment, which proved to be less prone to nano-related interference issues compared to some traditional toxicity assays. Furthermore, this label-free and novel technique shows good potential for up-scaling in directions of an automated high-throughput screening and for future NM toxicity assessment. This work was supported by the EC FP7 NANoREG (Grant Agreement NMP4-LA-2013-310584), the Research Council of Norway, project NorNANoREG (239199/O70), the EuroNanoMed II 'GEMN' project (246672), and the UH-Nett Vest project.

Keywords: cytotoxicity, high-throughput, impedance, nanomaterials

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Authors: Subodh Kumar, Sanjeev Kumar Sharma, Gaurav Kaushik, Pramod Avti, Phulen Sarma, Bikash Medhi, Krishan Lal Khanduja

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Background: It is well known that cigarette smoke is one of the causative factors in various lung diseases especially cancer. Carcinogens and oxidant molecules present in cigarette smoke not only damage the cellular constituents (lipids, proteins, DNA) but may also regulate the molecular pathways involved in inflammation and cancer. Continuous oxidative stress caused by the constituents of cigarette smoke leads to higher PhospholipaseA₂ (PLA₂) activity, resulting in elevated levels of secondary metabolites whose role is well defined in cancer. To reduce the burden of chronic inflammation as well as oxidative stress, and higher levels of secondary metabolites, we checked the curative potential of PLA₂ inhibitor Bromoenol Lactone (BEL) during continuous exposure of cigarette smoke condensate (CSC). Aim: To check the therapeutic potential of Bromoenol Lactone (BEL), an inhibitor of PhospholipaseA₂s, in pathways of CSC-induced changes in type I and type II alveolar epithelial cells. Methods: Effect of BEL on CSC-induced PLA2 activity were checked using colorimetric assay, cellular toxicity using cell viability assay, membrane integrity using fluorescein di-acetate (FDA) uptake assay, reactive oxygen species (ROS) levels and apoptosis markers through flow cytometry, and cellular regulation using MAPKinases levels, in lung epithelium. Results: BEL significantly mimicked CSC-induced PLA₂ activity, ROS levels, apoptosis, and kinases level whereas improved cellular viability and membrane integrity. Conclusions: Current observations revealed that BEL may be a potential therapeutic agent during Cigarette smoke-induced anomalies in lung epithelium.

Keywords: cigarette smoke condensate, phospholipase A₂, oxidative stress, alveolar epithelium, bromoenol lactone

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Authors: Mustafa Albayrak, Mete Onur Kaman, Ilyas Bozkurt

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In this study, the necessary material parameters were determined to be able to conduct progressive damage analysis of layered composites under low velocity impact by using the MAT162 material module in the LS-DYNA program. The material module MAT162 based on Hashin failure criterion requires 34 parameters in total. Some of these parameters were obtained directly as a result of dynamic and quasi-static mechanical tests, and the remaining part was calibrated and determined by comparing numerical and experimental results. Woven glass/epoxy was used as the composite material and it was produced by vacuum infusion method. In the numerical model, composites are modeled as three-dimensional and layered. As a result, the acquisition of MAT162 material module parameters, which will enable progressive damage analysis, is given in detail and step by step, and the selection methods of the parameters are explained. Numerical data consistent with the experimental results are given in graphics.

Keywords: Composite Impact, Finite Element Simulation, Progressive Damage Analyze, LS-DYNA, MAT162

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Authors: Xiran Wang, Johanna Trinkl, Thomas Hoffmann, Wilfried Schwab

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Malonyltransferases (MATs) are enzymes that play a key role in the biosynthesis of secondary metabolites in plants, such as flavonoids and anthocyanins. As a kind of flavonoid-rich fruit, strawberries are an ideal model to study MATs. From Goodberry metabolome data, in the hybrid generation of 2 strawberries various, Fragaria × ananassa cv. 'Senga Sengana' and 'Candonga', we found the malonylated flavonoid concentration is significantly higher in 'Senga Sengana' compared with 'Candonga'. Therefore, we aimed to identify and characterize the malonyltransferases responsible for the different malonylated flavonoid concentrations in two different strawberry cultivars. In this study, we have found 6 MATs via genome mapping, metabolome analysis, gene cloning, and enzyme assay from strawberries, which catalyzed the malonylation of flavonoid substrates: quercetin-3-glucoside, kaempferol-3-glucoside, pelargonidin-3-glucoside, and cyanidin-3-glucoside. All four compounds reacted with FaMATs to varying degrees. These MATs have important implication into strawberries’ flavonoid biosynthesis, and also provide insights into insights into flavonoid biosynthesis, potential applications in agriculture, plant science, and pharmacy, and information on the regulation of secondary metabolism in plants.

Keywords: malonyltransferase, strawberry, flavonoid biosynthesis, enzyme assay

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Authors: Mehmet Bozca

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Singular value decomposition based optimisation of geometric design parameters of a 5-speed gearbox is studied. During the optimisation, a four-degree-of freedom torsional vibration model of the pinion gear-wheel gear system is obtained and the minimum singular value of the transfer matrix is considered as the objective functions. The computational cost of the associated singular value problems is quite low for the objective function, because it is only necessary to compute the largest and smallest singular values (µmax and µmin) that can be achieved by using selective eigenvalue solvers; the other singular values are not needed. The design parameters are optimised under several constraints that include bending stress, contact stress and constant distance between gear centres. Thus, by optimising the geometric parameters of the gearbox such as, the module, number of teeth and face width it is possible to obtain a light-weight-gearbox structure. It is concluded that the all optimised geometric design parameters also satisfy all constraints.

Keywords: Singular value, optimisation, gearbox, torsional vibration

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Authors: G. M. Hamada, A. A. Al-Gathe, A. M. Al-Khudafi

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Determination of water saturation in sandstone is a vital question to determine the initial oil or gas in place in reservoir rocks. Water saturation determination using electrical measurements is mainly on Archie’s formula. Consequently accuracy of Archie’s formula parameters affects water saturation values rigorously. Determination of Archie’s parameters a, m, and n is proceeded by three conventional techniques, Core Archie-Parameter Estimation (CAPE) and 3-D. This work introduces the hybrid system of parallel self-organizing neural network (PSONN) targeting accepted values of Archie’s parameters and, consequently, reliable water saturation values. This work focuses on Archie’s parameters determination techniques; conventional technique, CAPE technique, and 3-D technique, and then the calculation of water saturation using current. Using the same data, a hybrid parallel self-organizing neural network (PSONN) algorithm is used to estimate Archie’s parameters and predict water saturation. Results have shown that estimated Arche’s parameters m, a, and n are highly accepted with statistical analysis, indicating that the PSONN model has a lower statistical error and higher correlation coefficient. This study was conducted using a high number of measurement points for 144 core plugs from a sandstone reservoir. PSONN algorithm can provide reliable water saturation values, and it can supplement or even replace the conventional techniques to determine Archie’s parameters and thereby calculate water saturation profiles.

Keywords: water saturation, Archie’s parameters, artificial intelligence, PSONN, sandstone reservoir

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Authors: Mmbulaheni Ramulondi, Sandy Van Vuuren, Helene De Wet

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The use of plant combinations to treat various medical conditions is not a new concept, and it is known that traditional people do not only rely on a single plant extract for efficacy but often combine various plant species for treatment. The knowledge of plant combinations is transferred from one generation to the other in the belief that combination therapy may enhance efficacy, reduce toxicity, decreases adverse effects, increase bioavailability and result in lower dosages. However, combination therapy may also be harmful when the interaction is antagonistic, since it may result in increasing toxicity. Although a fair amount of research has been done on the toxicity of medicinal plants, there is very little done on the toxicity of medicinal plants in combination. The aim of the study was to assess the toxicity potential of 19 plant combinations which have been documented as treatments of hypertension in northern KwaZulu-Natal by lay people. The aqueous extracts were assessed using two assays; the Brine shrimp assay (Artemia franciscana) and the Ames test (Mutagenicity). Only one plant combination (Aloe marlothii with Hypoxis hemerocallidea) in the current study has been previously assessed for toxicity. With the Brine shrimp assay, the plant combinations were tested in two concentrations (2 and 4 mg/ml), while for mutagenicity tests, they were tested at 5 mg/ml. The results showed that in the Brine shrimp assay, six combinations were toxic at 4 mg/ml. The combinations were Albertisia delagoensis with Senecio serratuloides (57%), Aloe marlothii with Catharanthus roseus (98%), Catharanthus roseus with Hypoxis hemerocallidea (66%), Catharanthus roseus with Musa acuminata (89%), Catharanthus roseus with Momordica balsamina (99%) and Aloe marlothii with Trichilia emetica and Hyphaene coriacea (50%). However when the concentration was reduced to 2 mg/ml, only three combinations were toxic which were Aloe marlothii with Catharanthus roseus (76%), Catharanthus roseus with Musa acuminata (66%) and Catharanthus roseus with Momordica balsamina (73%). For the mutagenicity assay, only the combinations between Catharanthus roseus with Hypoxis hemerocallidea and Catharanthus roseus with Momordica balsamina were mutagenic towards the Salmonella typhimurium strains TA98 and TA100. Most of the combinations which were toxic involve C. roseus which was also toxic when tested singularly. It is worth noting that C. roseus was one of the most frequently used plant species both to treat hypertension singularly and in combination and some of the individuals have been using this for the last 20 years. The mortality percentage of the Brine shrimp showed a significant correlation between dosage and toxicity thus toxicity was dosage dependant. A combination which is worth noting is the combination between A. delagoensis and S. serratuloides. Singularly these plants were non-toxic towards Brine shrimp, however their combination resulted in antagonism with the mortality rate of 57% at the total concentration of 4 mg/ml. Low toxicity was mostly observed, giving some validity to combined use, however the few combinations showing increased toxicity demonstrate the importance of analysing plant combinations.

Keywords: dosage, hypertension, plant combinations, toxicity

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Authors: Mondal Indranil, Bhakat Debjyoti, Mukhopadayay Asish K., Chatterjee Nabendu S.

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CS6 is the prevalent CF in our region and deciphering its molecular regulators would play a pivotal role in reducing the burden of ETEC pathogenesis. In prokaryotes, most of the genes are under the control of one operon and the promoter present upstream of the gene regulates the transcription of that gene. Here the promoter of CS6 was characterized by computational method and further analyzed by β-galactosidase assay and sequencing. Promoter constructs and deletions were prepared as required to analyze promoter activity. The effect of different additives on the CS6 promoter was analysed by the β-galactosidase assay. Bioinformatics analysis done by Softberry/BPROM predicted fur, lrp, and crp boxes, -10 and -35 region upstream of the CS6 gene. The promoter construction in no promoter plasmid pTL61T showed that region -573 to +1 is actually the promoter region as predicted. Sequential deletion of the region upstream of CS6 revealed that promoter activity remains the same when -573bp to -350bp is deleted. But after the deletion of the upstream region -350 bp to -255bp, promoter expression decreases drastically to 26%. Further deletion also decreases promoter activity up to a little range. So the region -355bp to -255bp holds the promoter sequence for the CS6 gene. Additives like iron, NaCl, etc., modulate promoter activity in a dose-dependent manner. From the promoter analysis, it can be said that the minimum region lies between -254 and +1. Important region(s) lies between -350 bp to -255 bp upstream in the promoter, which might have important elements needed to control CS6 gene expression.

Keywords: microbiology, promoter, colonization factor, ETEC

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Authors: I. Toumi, Y. Abed, A. Bouafia

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This paper presents a methodology for identifying the cohesive soil parameters that takes into account different constitutive equations. The procedure, applied to identify the parameters of generalized Prager model associated to the Drucker & Prager failure criterion from a pressuremeter expansion curve, is based on an inverse analysis approach, which consists of minimizing the function representing the difference between the experimental curve and the simulated curve using a simplex algorithm. The model response on pressuremeter path and its identification from experimental data lead to the determination of the friction angle, the cohesion and the Young modulus. Some parameters effects on the simulated curves and stresses path around pressuremeter probe are presented. Comparisons between the parameters determined with the proposed method and those obtained by other means are also presented.

Keywords: cohesive soils, cavity expansion, pressuremeter test, finite element method, optimization procedure, simplex algorithm

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Authors: Aakansha Gupta, Neha Vadnere, Tapasvi Soni, M. Anbarsi

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Protein secondary structure prediction is one of the most important goals pursued by bioinformatics and theoretical chemistry; it is highly important in medicine and biotechnology. Some aspects of protein functions and genome analysis can be predicted by secondary structure prediction. This is used to help annotate sequences, classify proteins, identify domains, and recognize functional motifs. In this paper, we represent protein secondary structure as a mathematical model. To extract and predict the protein secondary structure from the primary structure, we require a set of parameters. Any constants appearing in the model are specified by these parameters, which also provide a mechanism for efficient and accurate use of data. To estimate these model parameters there are many algorithms out of which the most popular one is the EM algorithm or called the Expectation Maximization Algorithm. These model parameters are estimated with the use of protein datasets like RS126 by using the Bayesian Probabilistic method (data set being categorical). This paper can then be extended into comparing the efficiency of EM algorithm to the other algorithms for estimating the model parameters, which will in turn lead to an efficient component for the Protein Secondary Structure Prediction. Further this paper provides a scope to use these parameters for predicting secondary structure of proteins using machine learning techniques like neural networks and fuzzy logic. The ultimate objective will be to obtain greater accuracy better than the previously achieved.

Keywords: model parameters, expectation maximization algorithm, protein secondary structure prediction, bioinformatics

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Authors: Eva Tvrdá, Hana Greifová, Peter Ivanič, Norbert Lukáč

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The aim of this study was to evaluate the dose- and time-dependent in vitro effects of berberine (BER), a natural alkaloid with numerous biological properties on bovine spermatozoa during three time periods (0 h, 2 h, 24 h). Bovine semen samples were diluted and cultivated in physiological saline solution containing 0.5% DMSO together with 200, 100, 50, 10, 5, and 1 μmol/L BER. Spermatozoa motility was assessed using the computer assisted semen analyzer. The viability of spermatozoa was assessed by the metabolic (MTT) assay, production of superoxide radicals was quantified using the nitroblue tetrazolium (NBT) test, and chemiluminescence was used to evaluate the generation of reactive oxygen species (ROS). Cell lysates were prepared and the extent of lipid peroxidation (LPO) was evaluated using the TBARS assay. The results of the movement activity showed a significant increase in the motility during long term cultivation in case of concentrations ranging between 1 and 10 μmol/L BER (P < 0.01; P < 0.001; 24 h). At the same time, supplementation of 1, 5 and 10 μmol/L BER led to a significant preservation of the cell viability (P < 0.001; 24 h). BER addition at a range of 1-50 μmol/L also provided a significantly higher protection against superoxide (P < 0.05) and ROS (P < 0.001; P < 0.01) overgeneration as well as LPO (P < 0.01; P<0.05) after a 24 h cultivation. We may suggest that supplementation of BER to bovine spermatozoa, particularly at concentrations ranging between 1 and 50 μmol/L, may offer protection to the motility, viability and oxidative status of the spermatozoa, particularly notable at 24 h.

Keywords: berberine, bulls, motility, oxidative profile, spermatozoa, viability

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Authors: Muhammad Farooq Baig

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Objective: The purpose of this study was to compare results obtained using a single fecal specimen for O&P examination, direct immunofluorescence assay (DFA), and two conventional staining methods. Design: Hundred and fifty children fecal specimens were collected and examined by each method. The O&P and the DFA were used as the reference method. Setting: The study was performed at the laboratory in the Basic Medical Science Institute JPMC Karachi. Patients or Other Participants: The fecal specimens were collected from children with a suspected Giardia lamblia infection. Main Outcome Measures: The amount of agreement and disagreement between methods.1) Presence of giardiasis in our population. 2) The sensitivity and specificity of each method. Results: There was 45(30%) positive 105 (70%) negative on DFA, 41 (27.4%) positive 109 (72.6%) negative on iodine and 34 (22.6%) positive 116(77.4%) on saline method. The sensitivity and specificity of DFA in comparision to iodine were 92.2%, 92.7% respectively. The sensitivity and specificity of DFA in comparisoin to saline method were 91.2%, 87.9% respectively. The sensitivity of iodine method and saline method in compariosn to DFA were 82.2%, 68.8% respectively. There is mark diffrence in sensitivity of DFA to conventional method. Conclusion: The study supported findings of other investigators who concluded that DFA method have the greater sensitivity. The immunologic methods were more efficient and quicker than the conventional O&P method.

Keywords: direct immunofluorescence assay (DFA), ova and parasite (O&P), Giardia lamblia, children, medical science

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Authors: Giancarlo La Marca, Engy Shokry, Fabio Villanelli

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Epilepsy is one of the most common neurological disorders, with an estimated prevalence of 50 million people worldwide. Twenty five percent of the epilepsy population is represented in children under the age of 15 years. For antiepileptic drugs (AED), there is a poor correlation between plasma concentration and dose especially in children. This was attributed to greater pharmacokinetic variability than adults. Hence, therapeutic drug monitoring (TDM) is recommended in controlling toxicity while drug exposure is maintained. Carbamazepine (CBZ) is a first-line AED and the drug of first choice in trigeminal neuralgia. CBZ is metabolised in the liver into carbamazepine-10,11-epoxide (CBZE), its major metabolite which is equipotent. This develops the need for an assay able to monitor the levels of both CBZ and CBZE. The aim of the present study was to develop and validate a LC-MS/MS method for simultaneous quantification of CBZ and CBZE in dried blood spots (DBS). DBS technique overcomes many logistical problems, ethical issues and technical challenges faced by classical plasma sampling. LC-MS/MS has been regarded as superior technique over immunoassays and HPLC/UV methods owing to its better specificity and sensitivity, lack of interference or matrix effects. Our method combines advantages of DBS technique and LC-MS/MS in clinical practice. The extraction process was done using methanol-water-formic acid (80:20:0.1, v/v/v). The chromatographic elution was achieved by using a linear gradient with a mobile phase consisting of acetonitrile-water-0.1% formic acid at a flow rate of 0.50 mL/min. The method was linear over the range 1-40 mg/L and 0.25-20 mg/L for CBZ and CBZE respectively. The limit of quantification was 1.00 mg/L and 0.25 mg/L for CBZ and CBZE, respectively. Intra-day and inter-day assay precisions were found to be less than 6.5% and 11.8%. An evaluation of DBS technique was performed, including effect of extraction solvent, spot homogeneity and stability in DBS. Results from a comparison with the plasma assay are also presented. The novelty of the present work lies in being the first to quantify CBZ and its metabolite from only one 3.2 mm DBS disc finger-prick sample (3.3-3.4 µl blood) by LC-MS/MS in a 10 min. chromatographic run.

Keywords: carbamazepine, carbamazepine-10, 11-epoxide, dried blood spots, LC-MS/MS, therapeutic drug monitoring

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Authors: Cruz, G. P., Nagamatsu, R. N., Scacchetti, F. A. P., Merlin, F. K.

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This study aims to assess a set of technical parameters that provide quality products to the companies that produce caps, APL Apucarana / PR, the city that produces most Brazilian caps, in order to verify the potential of Brazilian caps to compete with international brands, recognized by the standard of excellence when it comes to quality of its products. The determination of the technical parameters was arbitrated from textile ABNT, a total of six technical parameters, providing eight tests for cotton caps. For the evaluation, we used as reference a leading brand recognized worldwide (based on their sales volume in $) for comparison with 3 companies of the APL Apucarana. The results showed that, of the 8 tests, of 8 tests, the companies Apucarana did not obtain better performance than the competitor. They obtained the same results in three tests and lower performance in 5. Given these values, it is concluded that local caps are not far from reaching the quality of leading brand. It is recommended that the APL companies use the parameters to evaluate their products, using this information to support decision-making that seek to improve both the product design and its production process, enabling the feasibility for faster international recognition . Thus, they may have an edge over its main competitor.

Keywords: technical parameters, making caps, quality, evaluation

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Authors: P. H. Mfengwana, S. S. Mashele, L. Verschaeve, R. Anthonissen, I. T. Manduna

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Gunnera perpensa is traditionally used mostly by women for the treatment of different gynaecological related conditions due to its proven uterine contractility effects. The uses of this plant include menstrual pain relief, treatment of infertility and promotion of easy labour. However, even though this plant species has been reported to possess numerous medicinal properties, to author’s best knowledge, its safety has not been investigated. Thus, this study was aimed at investigating the genotoxicity and antigenotoxicity of Gunnera perpensa aqueous, methanol and dichloromethane extracts. The in vitro toxicity of the plant extracts was assessed with the neutral red uptake (NRU) test. Genotoxic and antigenotoxic properties of Gunnera perpensa were investigated using high-throughput assays: bacterial Vitotox test and the alkaline comet assay with and without S9 activation on human C3A cells. Ethyl Methanesulfonate (EMS) and 4-nitroquinoline-oxide (4-NQO) were used as positive controls, respectively. All extracts showed toxicity in a dose-dependent manner; however, that does not mean they were all genotoxic. Methanol extract did show genotoxicity with S9 (metabolism) only at the highest concentration of 500 µg/ml due to increased DNA damage observed, however, no genotoxicity was observed from other concentrations. Therefore, the results show that Gunnera perpensa extracts are genotoxic and not safe for human use.

Keywords: antigenotoxicity, comet test, genotoxicity, Gunnera perpensa, vitotox assay

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Authors: Shazia Naheed Akhter, Rekha Kumari

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An estimated 70 million population are exposed to arsenic poisoning in India in recent times. Arsenic contamination in the groundwater has caused serious health hazards among the exposed population. In Bihar, the first district was Bhojpur, where arsenic causing health issues were reported in 2002. Presently, there are 18 districts that are reported arsenic poisoning in the groundwater. The exposed population is firstly diseased with various symptoms such as skin manifestations, loss of appetite, constipation, hormonal disorders, etc. The long duration exposure has led to cause infertility in the male subjects. The present study thus aims to develop the antidote against arsenic-induced male reproductive toxicity in animal models. The study was carried out on Charles Foster Rats after the approval from Institutional Animal Ethics Committee. A total of n=18 rats (12 weeks old) of an average weight of 160 ± 20 g were used for the study. The study group included n=6 control and n= 12 treated with sodium arsenite orally at the dose of 8mg/Kg b.w daily for 40 days. The n= 6 animals were dissected and the rest n=6 was administered orally with Curcuma longa rhizome ethanolic extract at the dose of 600mg/Kg b.w per day for 40 days. At the end of the entire experiment, all the animals were dissected out and their reproductive organs were taken out, especially epididymis for sperm counts, sperm motility, sperm mortality, sperm morphology. The blood samples were collected for the hormonal assay (testosterone and luteinizing hormone), as well as for hematological and biochemical analysis. The study showed a high magnitude of degeneration in the reproductive organs of the rats in the arsenic-treated group. There were degenerative fluctuations in the sperm counts, sperm motility, sperm mortality, sperm morphology and in the hormonal parameters, as well as in the hematological and biochemical parameters in the arsenic-treated rats. But, after the administration of Curcuma longa, there was significant amelioration in all these parameters. Therefore, the present study shows that Curcuma longa plays a vital role to combat arsenic-induced male reproductive toxicity.

Keywords: sodium arsenite, Charles foster rats, ethanolic rhizome extract of curcuma longa, male reproductive toxicity, amelioration

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Authors: Shabnam Abdi, Behzad Toosi

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Background: Melanoma is the most lethal type of malignant skin cancer in humans and dogs since it spreads rapidly throughout the body. Despite significant advances in treatment, cancer at an advanced stage has a poor prognosis. Hence, more effective treatments are needed to enhance outcomes with fewer side effects. Erythropoietin-producing hepatocellular receptors are the largest family of receptor tyrosine kinases and are divided into two subfamilies, EphA and EphB, both of which play a significant role in disease, especially cancer. Due to their association with proliferation and invasion in many aggressive types of cancer, Eph receptor tyrosine kinases (Eph RTKs) are promising cancer therapy molecules. Because these receptors have not been studied in canine melanoma, we investigated how EphA2 influences survival and tumorigenicity of melanoma cells. Methods: Expression of EphA2 protein in canine melanoma cell lines and human melanoma cell line was evaluated by Western blot. Melanoma cells were transduced with lentiviral particles encoding Eph-targeting shRNAs or non-silencing shRNAs (control) for silencing the expression of EphA2 receptor, and silencing was confirmed by Western blotting and immunofluorescence. The effect of siRNA treatment on cellular proliferation, colony formation, tumorsphere assay, invasion was analyzed by Resazurin assay Matrigel invasion assay, respectively. Results: Expression of EphA2 was detected in canine and human melanoma cell lines. Moreover, stably silencing EphA2 by specific shRNAs significantly and consistently decreased the expression of EphA2 protein in both human and canine melanoma cells. Proliferation, colony formation, tumorsphere and invasion of melanoma cells were significantly decreased in EphA2 siRNA-treated cells compared to control. Conclusion: Our data provide the first functional evidence that the EphA2 receptor plays a critical role in the malignant cellular behavior of melanoma in both human and dogs.

Keywords: ephA2, targeting, melanoma, human, canine

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Authors: Rafid Saeed Abdulrazak Alshkaki

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In this paper, zero-one inflated negative binomial distribution is considered, along with some of its structural properties, then its parameters were estimated using the method of moments. It is found that the method of moments to estimate the parameters of the zero-one inflated negative binomial models is not a proper method and may give incorrect conclusions.

Keywords: zero one inflated models, negative binomial distribution, moments estimator, non negative integer sampling

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Authors: Mohita Singh, Sunil Sachdev, Amrita Singh

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Background and Aim: Physical aerobic exercises promote positive changes in one’s mental health, intelligence, and ability to cope with stressful encounters. The present study was designed to explore the correlation between intelligence and stress parameters and to assess the correlation between the same parameters after the practice of one month of moderate-intensity physical exercise. Method: The study was conducted on thirty-five healthy male volunteer students to assess the correlation between stress parameters in subjects with varying level of general intelligence (GI) and emotional intelligence (EI). Correlation studies were again conducted after one month between the same parameters to evaluate the effect of moderate-intensity physical exercise (MIPE). Baseline values were recorded using standard scales. Result: IQ and EQ correlated negatively with both acute and chronic stress parameters and positively with each other. A positive correlation was found between acute and chronic stress. With the practice of one month of moderate-intensity physical exercise, there was significant increment between the parameters under study and hence improved results. Conclusion: MIPE improved correlation between GI, EI, stress parameters, and thus reduced stress and improved intelligence.

Keywords: emotional intelligence, general intelligence, moderate intensity physical exercise, stress response

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Authors: Vasudha Devi, Arul Amuthan, K. Narayanan, Praveen KS, Venkata Rao J

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Background: Siddha Medicine is one of the Indian traditional medical systems, in which the cancer disease is mentioned as 'putrunoi' which literally means the disease of growth like termite mound. There are number of formulations available for the treatment of cancer disease. Neeradi muthu vallathymezugu (NMV) and thamira kattu chendooram (TKC) are two drugs commonly prescribed by Siddha physicians. These drugs have been clinically reported to be safe and effective when given orally. Though these formulations are in practice for centuries, no efforts have been made to standardize them and explore their anti-cancer potential systematically. Objective: To compare the cytotoxic activity of NMV and TKC with doxorubicin using cancer cell lines. Materials and methods: For this study, ethanol extract of NMV was taken, whereas TKC was used as such. In vitro cytotoxic activity was evaluated by sulphorhodamine (SRB) assay against human hepatic cancer cells (HepG2), human breast cancer cells (MCF-7) and human cervical cancer cells [KeLa]. Doxorubicin was used as the standard. The SRB assay is based on the ability of cellular proteins to bind with sulphorhodamine-B. The number of live cells in drug treated cell lines directly affects the color formation in the assay, which is estimated calorimetrically by measuring the absorbance at 540 nm to calculate the cytotoxicity (inhibitory concentration - IC50 value) of the drug. Results: The IC50values of NMV, TKC and doxorubicin against HepG2 were 3.08 µg/ml, 20.21 µg/ml and 1.21µg/ml respectively. In MCF-7, it was 11.75 µg/ml, 17.67 µg/ml and 2.8µg/ml. In HeLa, the values were 24.76 µg/ml, 73.35 µg/ml and 1.12µg/ml. Conclusions: The study proves the possible anti-cancer potential of these two formulations. Compared to TKC, NMV showed good cytotoxic effect even at low dose. Human hepatic cancer cells responded well even at very low dose, when compared to other cancer cells. Though, cytotoxic potential of these compounds was found to be less compared to doxorubicin, the isolated lead compound may have the potential to be used as an anticancer drug clinically.

Keywords: Neeradi muthu vallathymezugu (Hydnocarpus laurifolia), thamira kattu chendooram, cytotoxicity, in-vitro, Siddha Medicine

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Authors: Cheng-Cao Sun, Shu-Jun Li, Cuili Yang, Yongyong Xi, Liang Wang, Feng Zhang, De-Jia Li

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Hsa-miRNA-326 (miR-326) has recently been discovered having anticancer efficacy in different organs. However, the role of miR-326 on non-small cell lung cancer (NSCLC) is still ambiguous. In this study, we investigated the role of miR-326 on the development of NSCLC. The results indicated that miR-326 was significantly down-regulated in primary tumor tissues and very low levels were found in NSCLC cell lines. Ectopic expression of miR-326 in NSCLC cell lines significantly suppressed cell growth as evidenced by cell viability assay, colony formation assay and BrdU staining, through inhibition of cyclin D1, cyclin D2, CDK4, and up-regulation of p57(Kip2) and p21(Waf1/Cip1). In addition, miR-326 induced apoptosis, as indicated by concomitantly with up-regulation of key apoptosis protein cleaved caspase-3, and down-regulation of anti-apoptosis protein Bcl2. Moreover, miR-326 inhibited cellular migration and invasiveness through inhibition of matrix metalloproteinases (MMP)-7 and MMP-9. Further, oncogene CCND1 was revealed to be a putative target of miR-326, which was inversely correlated with miR-326 expression in NSCLC. Taken together, our results demonstrated that miR-326 played a pivotal role on NSCLC through inhibiting cell proliferation, migration, invasion, and promoting apoptosis by targeting oncogenic CCND1.

Keywords: hsa-miRNA-326 (miR-326), cyclin D1, non-small cell lung cancer (NSCLC), proliferation, apoptosis

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Authors: Muhammad Naveed Aslam, Rida Fatima, Anam Moosa, Muhammad Taimoor Shakeel

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Bacillus strains produce antifungal secondary metabolites making them potential candidates for suppressing Fusarium wilt of chickpea disease. In this study, eighteen Bacillus strains were evaluated for their antagonistic effect against Fusarium oxysporum f. sp. ciceris causing Fusarium wilt of chickpea disease. In a direct antifungal assay, thirteen strains showed significant inhibition zones while the remaining five strains did not produce inhibition zones of FOC. Bacillus thuringiensis CHGP12 was the most promising strain exhibiting the highest inhibition of FOC. Antifungal lipopeptides were extracted from CHGP12 strain which showed significant inhibition of the pathogen. Liquid chromatography mass spectrometry (LCMS) analysis revealed that CHGP12 was positive for the presence of iturin, fengycin, surfactin, bacillaene, bacillibactin, plantazolicin, and bacilysin. CHGP12 was tested for biochemical determinants in an in vitro qualitative test where it showed the ability to produce lipase, amylase, cellulase, protease, siderophores, and indole 3-acetic acid (IAA). Furthermore, in a greenhouse experiment CHGP12 also showed a significant decrease in the disease severity in treated plants compared to control. Moreover, CHGP12 also exhibited a significant increase in plant growth parameters viz, root and shoot growth parameters, stomatal conductance, and photosynthesis rate. Conclusively, our findings present the promising potential of Bacillus strain CHGP12 to suppress Fusarium wilt of chickpea and to promote plant growth.

Keywords: liquid chromatography mass spectrometry, growth promotion, antagonism, hydrolytic enzymes, inhibition, lipopeptides.

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Authors: Churamani Pokhrel, C. B. Jha, S. R. Niraula, P. R. Pokharel

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Human identification is one of the most challenging subjects that man has confronted. The determination of adult sex and stature are two of the four key factors (sex, stature, age, and race) in identification of an individual. Craniofacial and odontometric parameters are important tools for forensic anthropologists when it is not possible to apply advanced techniques for identification purposes. The present study provides anthropometric correlation of the parameters with stature and gender and also devises regression formulae for reconstruction of stature. A total of 312 Nepalese students with equal distribution of sex i.e., 156 male and 156 female students of age 18-35 years were taken for the study. Total of 10 parameters were measured (age, sex, stature, head circumference, head length, head breadth, facial height, bi-zygomatic width, mesio-distal canine width and inter-canine distance of both maxilla and mandible). Co-relation and regression analysis was done to find the association between the parameters. All parameters were found to be greater in males than females and each was found to be statistically significant. Out of total 312 samples, the best regressor for the determination of stature was head circumference and mandibular inter-canine width and that for gender was head circumference and right mandibular teeth. The accuracy of prediction was 83%. Regression equations and analysis generated from craniofacial and odontometric parameters can be a supplementary approach for the estimation of stature and gender when extremities are not available.

Keywords: craniofacial, gender, odontometric, stature

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Authors: M. Pourshirazi, M. Esmaelifar, A. Aliahmadi, F. Yazdian, A. S. Hatamian Zarami, S. J. Ashrafi

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Medicinal fungi are the new potential source of drugs to improve the treatment of diseases with association to oxidative agents such as cancers. Monascus purpureus contains functional components potentially effective in improving human health. In the present work, ethanolic extract of Monascus purpureus (EEM) was evaluated for health improving potential mainly focusing on antioxidant and anticancer activities. Ferric ion reducing power (FRAP), scavenging of DPPH radicals and determining viability of breast carcinoma MCF-7 and cervical carcinoma HeLa cells with MTT assay were evaluated. Our data showed a significant antioxidant activity of EEM with 142.45 µg/ml inhibition concentration of 50% DPPH radicals and 2112.33 µg eq.Fe2+/mg extract of FRAP assay. These results might be caused by antioxidant components such as pigments and phenolic compounds. Further, the results demonstrated that EEM caused significant reduction in the viability of MCF-7 with IC50 of 7 µg/ml but not have good effect against viability of HeLa cells. Accordingly, Monascus purpureus is presented as a strong potential of breast cancer treatment. In further study, the mechanistic studies are needed to determine the mechanisms of anticancer activity of EEM.

Keywords: Monascus purpureus, antioxidant, cancer, ethanolic extract

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