Search results for: fraction ethyl acetate

Authors: Adebisi Adunola Demehin, Wanlaya Thamnarak, Jaruwan Chatwichien, Chatchakorn Eurtivong, Kiattawee Choowongkomon, Somsak Ruchirawat, Nopporn Thasana

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The epidermal growth factor receptor (EGFR) is a transmembrane glycoprotein involved in cellular signalling processes and, its aberrant activity is crucial in the development of many cancers such as lung cancer. Selaginellaceae are fern allies that have long been used in Chinese traditional medicine to treat various cancer types, especially lung cancer. Biflavonoids, the major secondary metabolites in Selaginellaceae, have numerous pharmacological activities, including anti-cancer and anti-inflammatory. For instance, amentoflavone induces a cytotoxic effect in the human NSCLC cell line via the inhibition of PARP-1. However, to the best of our knowledge, there are no studies on biflavonoids as EGFR inhibitors. Thus, this study aims to investigate the EGFR inhibitory activities of biflavonoids isolated from Selaginella siamensis and Selaginella bryopteris. Amentoflavone, tetrahydroamentoflavone, sciadopitysin, robustaflavone, robustaflavone-4-methylether, delicaflavone, and chrysocauloflavone were isolated from the ethyl-acetate extract of the whole plants. The structures were determined using NMR spectroscopy and mass spectrometry. In vitro study was conducted to evaluate their cytotoxicity against A549, HEPG2, and T47D human cancer cell lines using the MTT assay. In addition, a target-based assay was performed to investigate their EGFR inhibitory activity using the kinase inhibition assay. Finally, a molecular docking study was conducted to predict the binding modes of the compounds. Robustaflavone-4-methylether and delicaflavone showed the best cytotoxic activity on all the cell lines with IC50 (µM) values of 18.9 ± 2.1 and 22.7 ± 3.3 on A549, respectively. Of these biflavonoids, delicaflavone showed the most potent EGFR inhibitory activity with an 84% relative inhibition at 0.02 nM using erlotinib as a positive control. Robustaflavone-4-methylether showed a 78% inhibition at 0.15 nM. The docking scores obtained from the molecular docking study correlated with the kinase inhibition assay. Robustaflavone-4-methylether and delicaflavone had a docking score of 72.0 and 86.5, respectively. The inhibitory activity of delicaflavone seemed to be linked with the C2”=C3” and 3-O-4”’ linkage pattern. Thus, this study suggests that the structural features of these compounds could serve as a basis for developing new EGFR-TK inhibitors.

Keywords: anticancer, biflavonoids, EGFR, molecular docking, Selaginellaceae

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Authors: Yohannes Kelifa, Gomathi Periasamy, Aman Karim

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Introduction: Diabetes mellitus has become a major public health and economical problem across the globe. Modern antidiabetic drugs have a number of limitations, and scientific investigation of traditional herbal remedies used for diabetes may provide novel leads for the development of new antidiabetic drugs that can be used as alternative or complementary to available antidiabetic allopathic medications. Though Myrica salicifolia Hochst. ex A. Rich. is used for the management of diabetes in Ethiopian traditional medicine, there was no previous scientific evidence about its antidiabetic effect to the authors’ knowledge. This study was undertaken to evaluate the antidiabetic activity the root extracts of Myrica salicifolia in streptozotocin (STZ)-induced diabetic mice. Methods: Experimental diabetes was induced by intraperitoneal administration of STZ (150 mg/kg) in male mice. Diabetic mice were treated with oral doses of M. salicifolia root extracts at 200, 400 and 600 mg/kg, and its fractions (chloroform, ethyl acetate, n-butanol and aqueous) at a dose of 400 mg/kg daily for 15 days. Fasting blood glucose level (BGL) was measured at 0, 5th,10th, and 15th day. The free radical scavenging activity of the crude extract was determined using in vitro by DPPH assay. The statistical significance was assessed by one-way ANOVA, followed by Tukey’s multiple comparison tests. Results were considered significant when p < 0.05. Results: Daily administration of the M. salicifolia 80% methanol root extracts (at three different doses (200, 400 and 600 mg/kg) significantly (p < 0.05, p < 0.01 and p < 0.001) reduced fasting BGL compared with diabetic control. The aqueous and butanol fractions at a dose of 400 mg/kg resulted in maximum reduction of fasting BGL by 42.39%, and 52.13%, respectively at the 15th day in STZ-induced diabetic mice. Free radical scavenging activity of the 80% methanol extract of M. salicifolia was comparable to ascorbic acid. The IC50 values of the crude extract and ascorbic acid (a reference compound) were found to be 4.54 μg/ml and 4.39 μg/ml, respectively. Conclusion: These findings demonstrated that the methanolic extracts of M. salicifolia root and its fractions (n-butanol and aqueous) exhibit a significant antihyperglycemic activity in STZ-induced diabetic mice. Furthermore, the result of the present study indicates that M. salicifolia root extract is a potential source of natural antioxidants.

Keywords: antidiabetic, diabetes mellitus, DPPH, mice, Myrica salicifolia, streptozotocin

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Authors: Leelavinothan Pari , Ayyasamy Rathinam

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Diabetes mellitus (DM) is a major and growing public health problem throughout the world. Pterocarpus marsupium (Roxb.) (Family: Fabaceae) is widely used as a traditional medicine to treat various diseases including diabetes. However, the molecular mechanism of Pterocarpus marsupium has not been investigated so far. Two fractions (2.5% and 5%) of extract from the medicinal plant, Pterocarpus marsupium (PME) were conducted in a dose dependent manner in streptozotocin (45 mg/kg b.w.) induced type 2 diabetic rats. Each fraction of PME was administered to diabetic rats intragastrically at a dose of 50, 100 and 200 mg/kg b.w for 45 days. The effective dose 200 mg/kg b.w of 5% fraction was more pronounced in reducing the levels of blood glucose (95.65 mg/dL) and glycosylated hemoglobin (HbA1c) (0.41 mg/g Hb), and increasing the plasma insulin (16.20 µU/mL) level. Moreover, PME (200 mg/kg b.w) significantly ameliorated lipid peroxidation products (thiobarbituric reactive substances, lipid hydroperoxides) enzymatic (superoxide dismutase, catalase and glutathione peroxidase) and non-enzymatic antioxidants (Vitamin C, Vitamin E and reduced glutathione) levels. The altered activities of the key enzymes of lipid metabolism along with the lipid profile in diabetic rats were significantly reverted to near normal levels by the administration of PME 5% 200 mg/kg b.w fraction. PME (200 mg/kg b.w) has the ability to reduce the inflammatory cytokines, such as TNF-α, IL-6 mRNA, as well as protein expression and apoptotic marker, such as caspase-3 enzyme in diabetic hepatic tissue. The above biochemical findings were also supported by histological studies such as improvement in pancreas and liver. Pterocarpus marsupium could effectively reduce the hyperglycemia, oxidative-stress, inflammation and hyperlipedimea in diabetic rats; hence it could be a useful drug in the management of diabetes without any side effects.

Keywords: diabetes mellitus, streptozotocin, Pterocarpus marsupium, lipid peroxidation, Antioxidants, inflammatory cytokines

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Authors: Marcin Michalak

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Due to the increasing efforts on saving our natural environment a change in the structure of energy resources can be observed - an increasing fraction of a renewable energy sources. In many countries traditional underground coal mining loses its significance but there are still countries, like Poland or Germany, in which the coal based technologies have the greatest fraction in a total energy production. This necessitates to make an effort to limit the costs and negative effects of underground coal mining. The longwall complex is as essential part of the underground coal mining. The safety and the effectiveness of the work is strongly dependent of the diagnostic state of powered roof supports. The building of a useful and reliable diagnostic system requires a lot of data. As the acquisition of a data of any possible operating conditions it is important to have a possibility to generate a demanded artificial working characteristics. In this paper a new approach of modelling a leg pressure in the single unit of powered roof support. The model is a result of the analysis of a typical working cycles.

Keywords: machine modelling, underground mining, coal mining, structure

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Authors: Nageeb A. H. Haroun, Sabyasachi Mondal, Precious Sibanda

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The effects of thermal radiation, Soret and Dufour parameters on mixed convection and nanofluid flow over a stretching sheet in the presence of a magnetic field are investigated. The flow is subject to temperature dependent viscosity and a chemical reaction parameter. It is assumed that the nanoparticle volume fraction at the wall may be actively controlled. The physical problem is modelled using systems of nonlinear differential equations which have been solved numerically using a spectral relaxation method. In addition to the discussion on heat and mass transfer processes, the velocity, nanoparticles volume fraction profiles as well as the skin friction coefficient are determined for different important physical parameters. A comparison of current findings with previously published results for some special cases of the problem shows an excellent agreement.

Keywords: non-isothermal wedge, thermal radiation, nanofluid, magnetic field, soret and dufour effects

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Authors: Subeena Begum, Navaraj Perumalsamy

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The freshwater catfish Mystus keletius was injected with the methanolic extract (water soluble fraction) of Solanum trilobatum and Ocimum sanctum alone and in combination in 3mg, 30mg.300mg/Kg body weight. Serum was collected every 7 days interval. Fishes were fed with normal diet for the entire period of the experiment. The nonspecific immune response such as Total WBC count, phagocytic activity and serum antiprotease activity were observed. They were enhanced (p<0.05) in fish injected with methanolic extract (water soluble fraction) S.trilobatum and O. sanctum alone and in combination than control group. Highest level of WBC count, phagocytic activity and serum antiprotease activity were confirmed when the fish injected with the mixed extract of 1:1 ratio of S.trilobatum and O.sanctum in 30mg/kg (b.w).The effect of mixture of methanolic extract of medicinal plant in M. keletius is highlighted.

Keywords: Immunology, Fish, Stimulatory effect, Medicinal plants

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Authors: Willian L. G. Silva, Fabio R. M. Batista, Matthieu Tubino

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Microalgae can be considered a potential source of oil for biodiesel synthesis since this microorganism can grow rapidly in either fresh or salty water, not competing with food production. There are several favorable conditions in Brazil for this type of culture due to the country’s great amount of water. Another very positive aspect of this type of culture is its ability to fix atmospheric CO2, contributing to the reduction of greenhouse gases and their effects on global warming. Despite this biodiesel environmental advantages it degrades resulting in changes in its physical and chemical properties. In this work, the methyl and ethyl microalgae biodiesel oxidative stability was studied in the absence and presence of a synthetic antioxidant. The synthetic antioxidants used were propyl gallate (PG) and tert-butylhydroquinone (TBHQ), at a 0,12% (w/w) concentration. The biodiesel mixture was kept in a sealed glass flask, sheltered from light, and at room temperature (about 25 ºC) for 180 days. During this period, aliquots from this biodiesel were subjected to induced degradation by the Rancimat method, which determines an important quality parameter, provided in the current methods, and is used to monitor the degradation processes that occur in the biodiesel over time. The induction period (IP) expresses the biodiesel oxidative stability. It was stablished that the minimum accepted IP value for biodiesel is 8 hours. The results show that ethylic biodiesel increased its IP value from 7,6 hours to 31 hours when using PG, and to 67 hours when using TBHQ, exceeding the minimum accepted IP value. When the antioxidants were added to the methylic biodiesel samples, the IP was raised to 28 hours when using PG, and to 62 hours when using TBHQ. These values were maintained throughout the entire period of study (180 days). On the other hand, the biodiesel samples without additives maintained an IP above the allowed value for only 30 days. Therefore, in order to preserve microalgae biodiesel for longer periods of time, it is necessary to add antioxidants to both derivatives, i.e., the ethylic and methylic.

Keywords: biodiesel, microalgae, oxidative stability, storage, synthetic antioxidants

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Authors: Marinela Miclau, Melinda Vajda, Nicolae Miclau, Daniel Ursu

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Characterized by a simple structure, easy and low cost fabrication, the dye-sensitized solar cell (DSSC) attracted the interest of the scientific community as an attractive alternative of conventional Si-based solar cells and thin-film solar cells. Over the past 20 years, the main efforts have attempted to enhance the efficiency of n-type DSSCs, the highest efficiency record of 14.30% was achieved using the co-sensitization of two metal-free organic dyes and Co (II/III) tris(phenanthroline)-based redox electrolyte. In the last years, the development of the efficient p-type DSSC has become a research focus owing to the fact that the concept of tandem solar cell was proposed as the solution to increase the power conversion efficiency. A promising alternative for the photocathodes of p-type DSSC, cuprous (Cu2O) and cupric (CuO) oxides have been investigated because of its nontoxic nature, low cost, high natural abundance, a good absorption coefficient for visible light and a higher dielectric constant than NiO. In case of p-type DSSC based on copper oxides with I3-/I- as redox mediator, the highest conversion efficiency of 0.42% (Cu2O) and 0.03% (CuO) has achieved. Towards the increase in the performance, we have fabricated and analyzed the performance of p-type DSSC prepared with the binder-free porous Cu2O photocathodes. Porous thin film could be an attractive alternative for DSSC because of their large surface areas which enable the efficient absorption of the dyes and light. We propose a simple and one-step hydrothermal method for the preparation of porous Cu2O thin film using copper substrate, cupric acetate and ethyl cellulose. The cubic structure of Cu2O has been determined by X-ray diffraction (XRD) and porous morphology of thin film was emphasized by Scanning Electron Microscope Inspect S (SEM). Optical and Mott-Schottky measurements attest of the high quality of the Cu2O thin film. The binder-free porous Cu2O photocathode has confirmed the excellent photovoltaic properties, the best value reported for p-type DSSC (1%) in similar conditions being reached.

Keywords: cuprous oxide, dye-sensitized solar cell, hydrothermal method, porous photocathode

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Authors: Pamita Awasthi, Kirna, Shilpa Dogra, Manu Vatsal, Ritu Barthwal

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Doxorubicin, also known as adriamycin, is an anthracycline class of drug used in cancer chemotherapy. It is used in the treatment of non-Hodgkin’s lymphoma, multiple myeloma, acute leukemias, breast cancer, lung cancer, endometrium cancer and ovary cancers. It functions via intercalating DNA and ultimately killing cancer cells. The major side effects of doxorubicin are hair loss, myelosuppression, nausea & vomiting, oesophagitis, diarrhoea, heart damage and liver dysfunction. The minor modifications in the structure of compound exhibit large variation in the biological activity, has prompted us to carry out the synthesis of sulfonamide derivatives. Sulfonamide is an important feature with broad spectrum of biological activity such as antiviral, antifungal, diuretics, anti-inflammatory, antibacterial and anticancer activities. Structure of the synthesized compound N-(1-methyl-2-oxo-2-N-methyl anilino-ethyl)benzene sulfonamide confirmed by proton nuclear magnetic resonance (1H NMR),13C NMR, Mass and FTIR spectroscopic tools to assure the position of all protons and hence stereochemistry of the molecule. Further we have reported the binding potential of synthesized sulfonamide analogues in comparison to doxorubicin drug using Auto Dock 4.2 software. Computational binding energy (B.E.) and inhibitory constant (Ki) has been evaluated for the synthesized compound in comparison of doxorubicin against Poly (dA-dT).Poly (dA-dT) and Poly (dG-dC).Poly (dG-dC) sequences. The in vitro cytotoxic study against human breast cancer cell lines confirms the better anticancer activity of the synthesized compound over currently in use anticancer drug doxorubicin. The IC50 value of the synthesized compound is 7.12 µM where as for doxorubicin is 7.2 µ.

Keywords: Doxorubicin, auto dock, in silco, in vitro

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Authors: A. Lakshumu Naidu, K. Krishna Kishor

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This paper examines the results of an experimental study based on the engineering properties of banana peel reinforced epoxy composites. Experiments are carried out to study the effect of weight fraction on mechanical behavior of epoxy based polymer composites. The composites were made by varying the weight fraction of banana peel from 0 to 30% and banana peel were made using hand layup method. The fabricated composite samples were cut according to the ASTM standards for different experiments. Hardness test and density test were carried out at the samples. The maximum hardness, density, tensile strength, flexural strength and ILSS are getting for the material prepared with the 20 % reinforced banana peel epoxy composite. The detailed test results and observations are discussed sequentially in the paper.

Keywords: engineering properties, polymer, composite, mechanical behavior of banana peel

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Authors: Mohamed Hassan, Rahul Gajbhiye

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The high mobility and gravity override of CO₂ gas can be minimized by generating the CO₂ foam with the aid of surfactant. However, CO₂ is unable to generate the foam/stable foam above its supercritical point (1100 psi, 31°C). These difficulties with CO₂ foam is overcome by adding N₂ in small fraction to enhance the foam generation of CO₂ at supercritical conditions. This study shows how the addition of small quantity of N₂ helps in generating the CO₂ foam and performance of the CO₂/N₂ mixture foam in enhanced oil recovery. To investigate the performance of CO₂/N₂ foam, core-flooding experiments were conducted at elevated pressure and temperature condition (higher than supercritical CO₂ - 50°C and 1500 psi) in sandstone cores. Fluorosurfactant (FS-51) was used as a foaming agent, and n-decane was used as model oil in all the experiments. The selection of foam quality and N₂ fraction was optimized based on foam generation and stability tests. Every gas or foam flooding was preceded by seawater injection to simulate the behavior in the reservoir. The results from the core-flood experiments showed that the CO₂ and CO₂/N₂ foam flooding recovered an additional 34-40% of Original Initial Oil in Place (OIIP) indicating that foam flooding succeeded in producing more oil than pure CO₂ gas injection processes. Additionally, the performance CO₂/N₂ foam injection was better than CO₂ foam injection.

Keywords: CO₂/N₂ foam, enhanced oil recovery (EOR), supercritical CO₂, sweep efficiency

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Authors: E. M. Condori-Peñaloza, S. S. Costa

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Eleusine indica L, known as goose-grass, is considered a troublesome weed that can cause important economic losses in the agriculture worldwide. However, this grass is used as a medicinal plant in some regions of Brazil to treat influenza and pneumonia. In Africa and Asia, it is used to treat malaria and as diuretic, anti-helminthic, among other uses. Despite its therapeutic potential, little is known about the chemical composition and bioactive compounds of E. indica. Hitherto, two major flavonoids, schaftoside and vitexin, were isolated from aerial part of the species and showed inhibitory activity on lung neutrophil influxes in mice, suggesting a beneficial effect on airway inflammation. Therefore, the aim of this study was to analyze the chemical profile of aqueous extracts from aerial parts of Eleusine indica specimens using high performance liquid chromatography (HPLC-DAD) and 1H nuclear magnetic resonance spectroscopy (NMR), with emphasis on phenolic compounds. Specimens of E. indica were collected in Minas Gerais state, Brazil. Aerial parts of fresh plants were extracted by decoction (10% p/v). After spontaneous precipitation of the aqueous extract at 10-12°C for 24 hours, the supernatant obtained was frozen and lyophilized. After that, 1 g of the extract was dissolved into 25 mL of water and fractionated on a reverse phase chromatography column (RP-2), eluted with a gradient of H2O/EtOH. Five fractions were obtained. The extract and fractions had their chemical profile analyzed by using HPLC-DAD (C-18 column: 20 μL, 256 -365 nm; gradient water 0.01% phosphoric acid/ acetonitrile. The extract was also analyzed by NMR (1H, 500 MHz, D2O) in order to access its global chemical composition. HPLC-DAD analyses of crude extract allowed the identification of ten phenolic compounds. Fraction 1, eluted with 100% water, was poor in phenolic compounds and no major peak was detected. In fraction 2, eluted with 100% water, it was possible to observe one major peak at retention time (RT) of 23.75 minutes compatible with flavonoid; fraction 3, also eluted with 100% water, showed four peaks at RT= 21.47, 23.52, 24.33 and 25.84 minutes, all of them compatible with flavonoid. In fraction 4, eluted with 50%/ethanol/50% water, it was possible to observe 3 peaks compatible with flavonoids at RT=24.65, 26.81, 27.49 minutes, and one peak (28.83 min) compatible with a phenolic acid derivative. Finally, in fraction 5, eluted with 100% ethanol, no phenolic substance was detected. The UV spectra of all flavonoids detected were compatible with the flavone subclass (λ= 320-345 nm). The 1H NMR spectra of aerial parts extract showed signals in three regions: δ 0.8-3.0 ppm (aliphatic compounds), δ 3.0-5.5 ppm corresponding to carbohydrates (signals most abundant and overlapped), and δ 6.0-8.5 ppm (aromatic compounds). Signals compatible with flavonoids (rings A and B) could also be detected in the crude extract spectra. These results suggest the presence of several flavonoids in E. indica, which reinforces their therapeutic potential. The pharmacological activities of Eleusine indica extracts and fractions will be further evaluated.

Keywords: flavonoids, HPLC, NMR, phenolic compounds

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Authors: David Taillis, Oussama Becissa, Julien Gabaston, Jean-Michel Merillon, Tristan Richard, Stephanie Cluzet

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Nowadays, there is a strong pressure to reduce the phytosanitary inputs of synthetic chemistry in vineyards. It is, therefore, necessary to find viable alternatives in order to protect the vine against its major diseases. For this purpose, we suggest the use of a plant extract enriched in antimicrobial compounds. Being produced from vine trunks and roots, which are co-products of wine production, the extract produced is part of a circular economy. The antimicrobial molecules present in this plant material are polyphenols and, more particularly, stilbenes, which are derived from a common base, the resveratrol unit, and that are well known vine phytoalexins. The stilbenoids were extracted from trunks and roots (30/70, w/w) by a double extraction with ethyl acetate followed by enrichment by liquid-liquid extraction. The produced extract was characterized by UHPLC-MS, then its antimicrobial activities were tested on Plasmopara viticola and Botrytis cinerea in the laboratory and/or in greenhouse and in vineyard. The major compounds were purified, and their antimicrobial activity was evaluated on B. cinerea. Moreover, after its spraying, the effect of the stilbene extract on the plant defence status was evaluated by analysis of defence gene expression. UHPLC-MS analysis revealed that the extract contains 50% stilbenes with resveratrol, ε-viniferin and r-viniferin as major compounds. The extract showed antimicrobial activities on P. viticola with IC₅₀ and IC₁₀₀ respectively of 90 and 300 mg/L in the laboratory. In addition, it inhibited 40% of downy mildew development in greenhouse. However, probably because of the sensitivity of stilbenes to the environment, such as UV degradation, no activity has been observed in vineyard towards P. viticola development. For B. cinerea, the extract IC50 was 123 mg/L, with resveratrol and ε-viniferin being the most active stilbenes (IC₅₀ of 88 and 142 mg/L, respectively). The analysis of the expression of defence genes revealed that the extract can induce the expression of some defence genes 24, 48, and 72 hours after treatment, meaning that the extract has a defence-stimulating effect at least for the first three days after treatment. In conclusion, we produced a plant extract enriched in stilbenes with antimicrobial properties against two major grapevine pathogenic agents P. viticola and B. cinerea. In addition, we showed that this extract displayed eliciting activity of plant defences. This extract can therefore represent, after formulation development, a viable eco-friendly alternative for vineyard protection. Subsequently, the effect of the stilbenoid extract on primary metabolism will be evaluated by quantitative NMR.

Keywords: antimicrobial, bioprotection, grapevine, Plasmopara viticola, stilbene

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Authors: Jung-En Kuan, Whei-Fen Wu

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In this study, a symbiotic bacteria from yellow mealworm's (Tenebrio molitor) mid-gut was isolated with characteristics of growth on minimal-tributyrin medium. After a PCR-amplification of its 16s rDNA, the resultant nucleotide sequences were then analyzed by schemes of the phylogeny trees. Accordingly, it was designated as Pseudomonas nitroreducens D-01. Next, by searching the lipolytic enzymes in its protein data bank, one of those potential lipolytic α/β hydrolases was identified, again using PCR-amplification and nucleotide-sequencing methods. To construct an expression of this lipolytic gene in plasmids, the target-gene primers were then designed, carrying the C-terminal his-tag sequences. Using the vector pET21a, a recombinant lipolytic hydrolase D gene with his-tag nucleotides was successfully cloned into it, of which the lipolytic D gene is under a control of the T7 promoter. After transformation of the resultant plasmids into Eescherichia coli BL21 (DE3), an IPTG inducer was used for the induction of the recombinant proteins. The protein products were then purified by metal-ion affinity column, and the purified proteins were found capable of forming a clear zone on tributyrin agar plate. Shortly, its enzyme activities were determined by degradation of p-nitrophenyl ester(s), and the substantial yellow end-product, p-nitrophenol, was measured at O.D.405 nm. Specifically, this lipolytic enzyme efficiently targets p-nitrophenyl butyrate. As well, it shows the most reactive activities at 40°C, pH 8 in potassium phosphate buffer. In thermal stability assays, the activities of this enzyme dramatically drop when the temperature is above 50°C. In metal ion assays, MgCl₂ and NH₄Cl induce the enzyme activities while MnSO₄, NiSO₄, CaCl₂, ZnSO₄, CoCl₂, CuSO₄, FeSO₄, and FeCl₃ reduce its activities. Besides, NaCl has no effects on its enzyme activities. Most organic solvents decrease the activities of this enzyme, such as hexane, methanol, ethanol, acetone, isopropanol, chloroform, and ethyl acetate. However, its enzyme activities increase when DMSO exists. All the surfactants like Triton X-100, Tween 80, Tween 20, and Brij35 decrease its lipolytic activities. Using Lineweaver-Burk double reciprocal methods, the function of the enzyme kinetics were determined such as Km = 0.488 (mM), Vmax = 0.0644 (mM/min), and kcat = 3.01x10³ (s⁻¹), as well the total efficiency of kcat/Km is 6.17 x10³ (mM⁻¹/s⁻¹). Afterwards, based on the phylogenetic analyses, this lipolytic protein is classified to type IV lipase by its homologous conserved region in this lipase family.

Keywords: enzyme, esterase, lipotic hydrolase, type IV

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Authors: Salima Tabti, Chaima Maouche, Tinhinene Louaileche, Amel Djedouani, Ismail Warad

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Novel Schiff base ligand HL has been synthesized by condensation of aromatic amine and DHA. It was characterized by UV-Vis, FT-IR, SM, NMR (1H, 13C) and also by single-crystal X-ray diffraction. The crystal structure shows that compound crystallized in a triclinic system in P-1 space group and with a two unit per cell (Z = 2).The asymmetric unit, contains one independent molecules, the conformation is determined by an intermolecular N-H…O hydrogen bond with an S(6) ring motif. The molecule have an (E) conformation about the C=N bond. The dihedral angles between the phenyl and pyran ring planes is 89.37 (1), the two plans are approximately perpendicular. The catecholase activity of is situ copper complexes of this ligand has been investigated against catechol. The progress of the oxidation reactions was closely monitored over time following the strong peak of catechol using UV-Vis. Oxidation rates were determined from the initial slope of absorbance. time plots, then analyzed by Michaelis-Menten equations. Catechol oxidation reactions were realized using different concentrations of copper acetate and ligand (L/Cu: 1/1, 1/2, 2/1). The results show that all complexes were able to catalyze the oxidation of catechol. Acetate complexes have the highest activity. Catalysis is a branch of chemical kinetics that, more generally, studies the influence of all physical or chemical factors determining reaction rates. It solves a lot of problems in the chemistry reaction process, especially for a green, economic and less polluting chemistry. For this reason, the search for new catalysts for known organic reactions, occupies a very advanced place in the themes proposed by the chemists.

Keywords: dehydroacetic acid, catechol, copper, catecholase activity, x-ray

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Authors: Selvam M., Vadthya Poornachandar, Surender Singh

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These Reclaimed Asphalt Pavement (RAP) aggregates are generally dumped in the open area after the demolition of Asphalt Pavements. The utilization of RAP aggregates in cement concrete pavements may provide several socio-economic-environmental benefits and could embrace the circular economy. The cross recycling of RAP aggregates in the concrete pavement could reduce the consumption of virgin aggregates and saves the fertile land. However, the structural, as well as functional properties of RAP-concrete could be significantly lower than the conventional Pavement Quality Control (PQC) pavements. This warrants judicious selection of RAP fraction (coarse and fine aggregates) along with the accurate proportion of the same for PQC highways. Also, the selection of the RAP fraction and its proportion shall not be solely based on the mechanical properties of RAP-concrete specimens but also governed by the structural and functional behavior of the pavement system. In this study, an effort has been made to predict the optimum RAP fraction and its corresponding proportion for cement concrete pavements by considering the low-volume and high-volume roads. Initially, the effect of inclusions of RAP on the fresh and mechanical properties of concrete pavement mixes is mapped through an extensive literature survey. Almost all the studies available to date are considered for this study. Generally, Indian Roads Congress (IRC) methods are the most widely used design method in India for the analysis of concrete pavements, and the same has been considered for this study. Subsequently, fatigue damage analysis is performed to evaluate the required safe thickness of pavement slab for different fractions of RAP (coarse RAP). Consequently, the performance of RAP-concrete is predicted by employing the AASHTO-1993 model for the following distresses conditions: faulting, cracking, and smoothness. The performance prediction and total cost analysis of RAP aggregates depict that the optimum proportions of coarse RAP aggregates in the PQC mix are 35% and 50% for high volume and low volume roads, respectively.

Keywords: concrete pavement, RAP aggregate, performance prediction, pavement design

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Authors: S. Serhal, Y. Melinge, D. Rangeard, F. Hage Chehadeh

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Filling of estuaries is an international problem that can cause economic and environmental damage. This work aims the study of the rheological structuring mechanisms of natural sedimentary liquid-solid mixture in estuaries in order to better understand their filling. The estuary of the Rance river, located in Brittany, France is particularly targeted by the study. The aim is to provide answers on the rheological behavior of natural sediments by detecting structural factors influencing the rheological parameters. So we can better understand the fillings estuarine areas and especially consider sustainable solutions of ‘cleansing’ of these areas. The sediments were collected from the trap of Lyvet in Rance estuary. This trap was created by the association COEUR (Comité Opérationnel des Elus et Usagers de la Rance) in 1996 in order to facilitate the cleansing of the estuary. It creates a privileged area for the deposition of sediments and consequently makes the cleansing of the estuary easier. We began our work with a preliminary study to establish the trend of the rheological behavior of the suspensions and to specify the dormant phase which precedes the beginning of the biochemical reactivity of the suspensions. Then we highlight the visco-plastic character at younger age using the Kinexus rheometer, plate-plate geometry. This rheological behavior of suspensions is represented by the Bingham model using dynamic yield stress and viscosity which can be a function of volume fraction, granular extent, and chemical reactivity. The evolution of the viscosity as a function of the solid volume fraction is modeled by the Krieger-Dougherty model. On the other hand, the analysis of the dynamic yield stress showed a fairly functional link with the solid volume fraction.

Keywords: estuaries, rheological behavior, sediments, Kinexus rheometer, Bingham model, viscosity, yield stress

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Authors: Oscar Corredor, Alexander Guzman, Adan Leon

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Density is a bulk physical property that indicates the quality of a petroleum fraction. It is also a useful property to estimate various physicochemical properties of fraction and petroleum fluids; however, the determination of density of extra heavy residual (EHR) fractions by standard methodologies, (ASTM D70) shows limitations for samples with higher densities than 1.0879 g/cm3. For this reason, a dilution methodology was developed in order to determinate density for those particular fractions, 87 (EHR) fractions were obtained as products of the fractionation of Colombian typical Vacuum Distillation Residual Fractions using molecular distillation (MD) and extraction with Solvent N-hexane in Supercritical Conditions (SFEF) pilot plants. The proposed methodology showed reliable results that can be demonstrated with the standard deviation of repeatability and reproducibility values of 0.0031 and 0.0061 g/ml respectively. In the same way, it was possible to determine densities in fractions EHR up to 1.1647g/cm3 and °API values obtained were ten times less than the water reference value.

Keywords: API, density, vacuum residual, molecular distillation, supercritical fluid extraction

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Authors: Morteza Taherinezhad, Ahmad Reza Rabbani, Morteza Asemani, Rudy Swennen

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Oil-oil correlation is one of the most important issues in geochemical studies that enables to classify oils genetically. Oil-oil correlation is generally estimated based on non-polar fractions of crude oil (e.g., saturate and aromatic compounds). Despite several advantages, the drawback of using these compounds is their susceptibility of being affected by secondary processes. The polar fraction of crude oil (e.g., asphaltenes) has similar characteristics to kerogen, and this structural similarity is preserved during migration, thermal maturation, biodegradation, and water washing. Therefore, these structural characteristics can be considered as a useful correlation parameter, and it can be concluded that asphaltenes from different reservoirs with the same genetic signatures have a similar origin. Hence in this contribution, an integrated study by using both non-polar and polar fractions of oil was performed to use the merits of both fractions. Therefore, five oil samples from oil fields in the Persian Gulf were studied. Structural characteristics of extracted asphaltenes were investigated by Fourier transform infrared (FTIR) spectroscopy. Graphs based on aliphatic and aromatic compounds (predominant compounds in asphaltenes structure) and sulphoxide and carbonyl functional groups (which are representatives of sulphur and oxygen abundance in asphaltenes) were used for comparison of asphaltenes structures in different samples. Non-polar fractions were analyzed by GC-MS. The study of asphaltenes showed the studied oil samples comprise two oil families with distinct genetic characteristics. The first oil family consists of Salman and Reshadat oil samples, and the second oil family consists of Resalat, Siri E, and Siri D oil samples. To validate our results, biomarker parameters were employed, and this approach completely confirmed previous results. Based on biomarker analyses, both oil families have a marine source rock, whereby marl and carbonate source rocks are the source rock for the first and the second oil family, respectively.

Keywords: biomarker, non-polar fraction, oil-oil correlation, petroleum geochemistry, polar fraction

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Authors: Edison E. Haro, Akindele G. Odeshi, Jerzy A. Szpunar

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Hybrid bio-composites are developed for use in protective armor through positive hybridization offered by reinforcement of high-density polyethylene (HDPE) with Kevlar short fibers and palm wood micro-fillers. The manufacturing process involved a combination of extrusion and compression molding techniques. The mechanical behavior of Kevlar fiber reinforced HDPE with and without palm wood filler additions are compared. The effect of the weight fraction of the added palm wood micro-fillers is also determined. The Young modulus was found to increase as the weight fraction of organic micro-particles increased. However, the flexural strength decreased with increasing weight fraction of added micro-fillers. The interfacial interactions between the components were investigated using scanning electron microscopy. The influence of the size, random alignment and distribution of the natural micro-particles was evaluated. Ballistic impact and dynamic shock loading tests were performed to determine the optimum proportion of Kevlar short fibers and organic micro-fillers needed to improve impact strength of the HDPE. These results indicate a positive hybridization by deposition of organic micro-fillers on the surface of short Kevlar fibers used in reinforcing the thermoplastic matrix leading to enhancement of the mechanical strength and dynamic impact behavior of these materials. Therefore, these hybrid bio-composites can be promising materials for different applications against high velocity impacts.

Keywords: hybrid bio-composites, organic nano-fillers, dynamic shocking loading, ballistic impacts, energy absorption

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Authors: Srinivasa Reddy Mallampati, Min Hee Park, Soo Mim Cho, Sung Hyeon Yoon

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One of the issues of End of Life Vehicles (ELVs) recycling promotion is technology for the appropriate treatment of automotive shredder residue (ASR). Owing to its high heterogeneity and variable composition (plastic (23–41%), rubber/elastomers (9–21%), metals (6–13%), glass (10–20%) and dust (soil/sand) etc.), ASR can be classified as ‘hazardous waste’, on the basis of the presence of heavy metals (HMs), PCBs, BFRs, mineral oils, etc. Considering their relevant concentrations, these metals and plastics should be properly recovered for recycling purposes before ASR residues are disposed of. Brominated flame retardant additives in ABS/HIPS and PVC may generate dioxins and furans at elevated temperatures. Moreover, these BFRs additives present in plastic materials may leach into the environment during landfilling operations. ASR thermal process removes some of the organic material but concentrates, the heavy metals and POPs present in the ASR residues. In the present study, Fe/Ca/CaO nanoparticle assisted ozone treatment has been found to selectively hydrophilize the surface of ABS/HIPS and PVC plastics, enhancing its wettability and thereby promoting its separation from ASR plastics by means of froth flotation. The water contact angles, of ABS/HIPS and PVC decreased, about 18.7°, 18.3°, and 17.9° in ASR respectively. Under froth flotation conditions at 50 rpm, about 99.5% and 99.5% of HIPS in ASR samples sank, resulting in a purity of 98% and 99%. Furthermore, at 150 rpm a 100% PVC separation in the settled fraction, with 98% of purity in ASR, respectively. Total recovery of non-ABS/HIPS and PVC plastics reached nearly 100% in the floating fraction. This process improved the quality of recycled ASR plastics by removing surface contaminants or impurities. Further, a hybrid ball-milling and with Fe/Ca/CaO nanoparticle froth flotation process was established for the recovery of HMs from ASR. After ball-milling with Fe/Ca/CaO nanoparticle additives, the flotation efficiency increased to about 55 wt% and the HMs recovery were also increased about 90% for the 0.25 mm size fractions of ASR. Coating with Fe/Ca/CaO nanoparticles associated with subsequent microbubble froth flotation allowed the air bubbles to attach firmly on the HMs. SEM–EDS maps showed that the amounts of HMs were significant on the surface of the floating ASR fraction. This result, along with the low HM concentration in the settled fraction, was confirmed by elemental spectra and semi-quantitative SEM–EDS analysis. Developed hybrid preferential hazardous plastics and metals separation process from ASR is a simple, highly efficient, and sustainable procedure.

Keywords: end of life vehicles shredder residue, hazardous plastics, nanoparticle froth flotation, separation process

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Authors: Ami Okabe, Daisuke Gondo, Akira Ogawa, Yasuhisa Hasegawa, Koichi Sato, Sadao Araki, Hideki Yamamoto

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Membrane separation draws attention as the energy-saving technology. Pervaporation (PV) uses hydrophobic ceramic membranes to separate organic compounds from industrial wastewaters. PV makes it possible to separate organic compounds from azeotropic mixtures and from aqueous solutions. For the PV separation of low concentrations of organics from aqueous solutions, hydrophobic ceramic membranes are expected to have high separation performance compared with that of conventional hydrophilic membranes. Membrane separation performance is evaluated based on the pervaporation separation index (PSI), which depends on both the separation factor and the permeate flux. Ingenuity is required to increase the PSI such that the permeate flux increases without reducing the separation factor or to increase the separation factor without reducing the flux. A thin separation layer without defects and pinholes is required. In addition, it is known that the flux can be increased without reducing the separation factor by reducing the diffusion resistance of the membrane support. In a previous study, we prepared hydrophobic silica membranes by a molecular templating sol−gel method using cetyltrimethylammonium bromide (CTAB) to form pores suitable for permitting the passage of organic compounds through the membrane. We separated low-concentration organics from aqueous solutions by PV using these membranes. In the present study, hydrophobic silica membranes were prepared on a porous alumina hollow fiber support that is thinner than the previously used alumina support. Ethyl acetate (EA) is used in large industrial quantities, so it was selected as the organic substance to be separated. Hydrophobic silica membranes were prepared by dip-coating porous alumina supports with a -alumina interlayer into a silica sol containing CTAB and vinyltrimethoxysilane (VTMS) as the silica precursor. Membrane thickness increases with the lifting speed of the sol in the dip-coating process. Different thicknesses of the γ-alumina layer were prepared by dip-coating the support into a boehmite sol at different lifting speeds (0.5, 1, 3, and 5 mm s-1). Silica layers were subsequently formed by dip-coating using an immersion time of 60 s and lifting speed of 1 mm s-1. PV measurements of the EA (5 wt.%)/water system were carried out using VTMS hydrophobic silica membranes prepared on -alumina layers of different thicknesses. Water and EA flux showed substantially constant value despite of the change of the lifting speed to form the γ-alumina interlayer. All prepared hydrophobic silica membranes showed the higher PSI compared with the hydrophobic membranes using the previous alumina support of hollow fiber.

Keywords: membrane separation, pervaporation, hydrophobic, silica

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Authors: Nurul Hamirah Kamsani, Mohd Hamim Rajikin, Nor Ashikin Mohamed Noor Khan, Sharaniza Abdul Rahim

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Cytoskeletal structures, in particular actin and tubulin, provide a fundamental framework in all cells, including embryos. Under influence of nicotine, the cytoskeletal organization may be subjected to oxidative stress (OS) insult and cause alteration. Tocotrienol-rich fraction (TRF) is proven to enhance fertility better than the other sub-group of Vitamin E, tocopherols (TCPs). The objective of this study was to evaluate the effects of TRF on 1) actin and tubulin of 2- and 8-cell murine embryos and 2) the regulation of reactive oxygen species (ROS)-scavenging enzymes; induced by nicotine. Twenty four female Balb/C were subjected to either subcutaneous (sc) injection of 0.9% NaCl; sc injection of 3.0 mg/kg bw/day nicotine; sc injection of 3.0 mg/kg bw/day nicotine + oral gavage (OG) of 60 mg/kg bw/day TRF; or OG of 60 mg/kg bw/day TRF for 7 consecutive days. After superovulation and mating, animals were euthanized. 2-cell developing embryos were retrieved. 50% of the retrieved embryos were visualized under confocal laser staining microscopy (CLSM) for alterations of actin and tubulin. The remaining amount of embryos was cultured in vitro until 8-cell stage followed by CLSM visualization. Blood plasma was subjected to OS assays. Plasma malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx) were determined and analysed accordingly. At both 2- and 8-cell developing stages, actin intensities were significantly reduced in the nicotine group (p<0.001). After the intervention, actin intensity was significantly increased compared to that of the nicotine group (p<0.001). The same trend was seen in tubulin at both cell stages. TRF has minimized the deleterious effects of nicotine in actin and tubulin of both 2- and 8-cell developmental stages during pre-implantation embryonic development in mice in vitro. Levels of endogenous anti-oxidative enzymes were sustained close to control accompanied by decreased levels of OS biomarker.

Keywords: actin, nicotine, pre-implantation embryos, tocotrienol rich fraction, tubulin

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Authors: Allulu Yousef Alturki, Raghad Abdullah Alshafi, Sara Abdulaziz Alghashem, Sahar Saleh Alghamdi, Rasha Saad Suliman, Zeyad Alehaideb, Rizwan Ali

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Cancer is recognized as a worldwide public health concern. Therefore, there is a continuous quest to discover new effective medications with less side-effects. In recent years, researchers have shown an increased interest in medicinal plants as several plant species have shown promising biological activities. Thus, we seek to investigate three medicinal herbs that are commonly-found in the Middle Easternregion and yet have not been explored in depth, including plants belonging to the Rhamnaceae, Polygonaceae, and Apocynaceaeplant families. Initially, we investigated using three types of cancer cell lines for breast, colorectal, and liver cancers. We performed high Content Imaging (HCI)-Apoptosis Assay and ApoTox-Glo™ Triplex Assay on KAIMRC2 and HCT8 cell lines. The highest activity of HCI-Apoptosis Assay was with Calligonumcomosum and Ziziphusnummularia in ethanol, followed by Calotropis procera and Ziziphusnummularia in ethyl acetate. The IC50values for the families of Rhamnaceae, Polygonaceae, and Apocynaceae in HepG2 and HCT8 cell lines ranged from 0.089 to 9.84mg/mL and 0.080to 15.08mg/mL, respectively. Further screening was conducted on an additional two cell lines, namely the MDA-MB-231 and KAIMRC2, for selected seven extracts with the highest activity having IC50values ranged from 0.058 to0.51mg/mL and 0.029 to0.19mg/mL, respectively. Continuous scientific investigations to isolate and characterize the potent bioactive phytochemical(s) are warranted. Funding: The authors acknowledge financial support from King Abdullah International Medical Research Center (KAIMRC), Ministry of National Guard Health Affairs, Riyadh, Kingdom of Saudi Arabia. Institutional Review Board Statement: The study was approved by the Institutional Review Board of the Institutional Review Board of King Abdullah International Medical Research Center (SP21R/463/12, 24 January 2022). Acknowledgments: The authors want to express their gratitude to the College of Pharmacy (COP) at King Saud bin Abdulaziz University for Health Sciences (KSAU-HS) and King Abdullah International Medical Research Center (KAIMRC) for their continued support.

Keywords: rhamnaceae, polygonaceae, apocynaceae, natural products

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Authors: Monika Kallubai, Shreya Dubey, Rajagopal Subramanyam

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Our study is all about the biological interactions of synthesized 5β-dihydrocortisol (Dhc) and 5β-dihydrocortisol acetate (DhcA) molecules with carrier protein Human Serum Albumin (HSA). The cytotoxic study was performed on breast cancer cell line (MCF-7) normal human embryonic kidney cell line (HEK293), the IC50 values for MCF-7 cells were 28 and 25 µM, respectively, whereas no toxicity in terms of cell viability was observed with HEK293 cell line. The further experiment proved that Dhc and DhcA induced 35.6% and 37.7% early apoptotic cells and 2.5%, 2.9% late apoptotic cells respectively. Morphological observation of cell death through TUNEL assay revealed that Dhc and DhcA induced apoptosis in MCF-7 cells. The complexes of HSA–Dhc and HSA–DhcA were observed as static quenching, and the binding constants (K) was 4.7±0.03×104 M-1 and 3.9±0.05×104 M-1, and their binding free energies were found to be -6.4 and -6.16 kcal/mol, respectively. The displacement studies confirmed that lidocaine 1.4±0.05×104 M-1 replaced Dhc, and phenylbutazone 1.5±0.05×104 M-1 replaced by DhcA, which explains domain I and domain II are the binding sites for Dhc and DhcA. Further, CD results revealed that the secondary structure of HSA was altered in the presence of Dhc and DhcA. Furthermore, the atomic force microscopy and transmission electron microscopy showed that the dimensions like height and molecular sizes of the HSA–Dhc and HSA–DhcA complex were larger compared to HSA alone. Detailed analysis through molecular dynamics simulations also supported the greater stability of HSA–Dhc and HSA–DhcA complexes, and root-mean-square-fluctuation interpreted the binding site of Dhc as domain IB and domain IIA for DhcA. This information is valuable for the further development of steroid derivatives with improved pharmacological significance as novel anti-cancer drugs.

Keywords: apoptosis, dihydrocortisol, fluorescence quenching, protein conformations

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Authors: Abbul B. Khan, Neeraj Dohare, Rajan Patel

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The mixed micellization of adiphenine hydrochloride (ADP) with 1-decyl-3-methylimidazolium chloride (C10mim.Cl), was investigated at different mole fractions and temperatures by surface tension measurements. The synergistic behavior (i.e., non-ideal behavior) for binary mixtures was explained by the deviation of critical micelle concentration (cmc) from ideal critical micelle concentration (cmc*), micellar mole fraction (Xim) from ideal micellar mole fraction (Xiideal), the values of interaction parameter (β) and activity coefficients (fi) (for both mixed micelles and mixed monolayer). The excess free energy (∆Gex) for the ADP- C10mim.Cl binary mixtures explain the stability of mixed micelles in comparison to micelles of pure ADP and C10mim.Cl. Interfacial parameters, i.e., Gibbs surface excess (Гmax), minimum head group area at air/ water interface (Amin), and free energy of micellization (ΔG0m) were also evaluated for the systems.

Keywords: adiphenine hydrochloride, critical micelle concentration, interaction parameter, activity coefficient

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Authors: Eyuphan Manay

Abstract:

In this study, it was aimed to determine the convective heat transfer characteristics of water-based silicon dioxide nanofluids (SiO₂) with particle volume fractions of 0.2 and 0.4% vol. Nanofluids were tested in a plate type heat exchanger with six plates. Plate type heat exchanger was manufactured from stainless steel. Water was driven in the hot flow side, and nanofluids were driven in the cold flow side. The thermal energy of the hot water was taken by nanofluids. Effect of the inlet temperature of the hot water was investigated on heat transfer performance of the nanofluids while the inlet temperature of the nanofluids was fixed. In addition, the effects of the particle volume fraction and the cold flow rate on the performance of the system were tested. Results showed that increasing inlet temperature of the hot flow caused heat transfer to enhance. The suspended solid particles into the carrier fluid also remarkably enhanced heat transfer, and, an increase in the particle volume fraction resulted in an increase in heat transfer.

Keywords: heat transfer enhancement, SiO₂-water, nanofluid, plate heat exchanger

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Authors: Jaber Masoud Alyami, Abdelsalam H. Alsrkhi

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Characterizing and modeling multi-phase flow is a complicated scientific and technical phenomenon represented by a variety of interrelated elements. Yet, the introduction of dimensionless numbers used to grasp gas-liquid flow is a significant step in controlling and improving the multi-phase flow area. SL (Slippage number), for instance is a strong dimensionless number defined as a the ratio of the difference in gravitational forces between slip and no-slip conditions to the inertial force of the gas. The fact that plotting SL versus Frm provides a single acceptable curve for all of the data provided proves that SL may be used to realize the behavior of gas-liquid flow. This paper creates a numerical link between SL and Froud mixing number using vertical gas-liquid flow and then utilizes that relationship to validate its reliability in practice. An improved correlation in drift flux model generated from the experimental data and its rationality has been verified. The method in this paper is to approach for predicting the void fraction in bubbly flow through SL/Frm relation and the limitations of this method, as well as areas for development, are stated.

Keywords: multiphase flow, gas-liquid flow, slippage, void farction

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Authors: M. Umar Dahot, G. S. Mangrio

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In this study Agro-industrial waste was used as a carbon source, which is a low cost substrate. Along with this, various sugars and molasses of 2.5% and 5% were investigated as substrate/carbon source for the growth of A.niger and Pectinase production. Different nitrogen sources were also used. An overview of results obtained show that 5% sucrose, 5% molasses and 0.4% (NH4)2SO4 were found the best carbon and nitrogen sources for the production of pectinase by A. niger. The maximum production of pectinase (26.87units/ml) was observed at pH 6.0 after 72 hrs incubation. The optimum temperature for the maximum production of pectinase was achieved at 35ºC when maximum production of pectinase was obtained as 28.25Units/ml.Pectinase enzyme was purified with ammonium sulphate precipitation and dialyzed sample was finally applied on gel filtration chromatography (Sephadex G-100) and Ion Exchange DEAE A-50. The enzyme was purified 2.5 fold by gel chromatography on Sephadex G-100 and Four fractions were obtained, Fraction 1, 2, 4 showed single band while Fraction -3 showed multiple bands on SDS Page electrophoresis. Fraction -3 was pooled, dialyzed and separated on Sephdex A-50 and two fractions 3a and 3b showed single band. The molecular weights of the purified fractions were detected in the range of 33000 ± 2000 and 38000± 2000 Daltons. The purified enzyme was specifically most active with pure pectin, while pectin, Lemon pectin and orange peel given lower activity as compared to (control). The optimum pH and temperature for pectinase activity was found between pH 5.0 and 6.0 and 40°- 50°C, respectively. The enzyme was stable over the pH range 3.0-8.0. The thermostability of was determined and it was observed that the pectinase activity is heat stable and retains activity more than 40% when incubated at 90°C for 10 minutes. The pectinase activity of F3a and F3b was increased with different metal ions. The Pectinase activity was stimulated in the presence of CaCl2 up to 10-30%. ZnSO4, MnSO4 and Mg SO4 showed higher activity in fractions F3a and F3b, which indicates that the pectinase belongs to metalo-enzymes. It is concluded that A. niger is capable to produce pH stable and thermostable pectinase, which can be used for industrial purposes.

Keywords: pectinase, a. niger, production, purification, characterization

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Authors: M. A. Munteanu, A. M. Lungu, A. I. Chivescu, V. Teodorescu, E. Tufanoiu, C. Nicolae, T. I. Nanea

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Introduction: Sodium-glucose co-transporter 2 inhibitors (SGLT2i), which were first developed as antidiabetic medications, have demonstrated numerous positive benefits on the cardiovascular system, especially in the prevention of heart failure (HF). HF is a challenging, multifaceted disease that needs all-encompassing therapy. It should not be viewed as a limited form of heart illness but rather as a systemic disease that leads to multiple organ failure and death. SGLT2i is an extremely effective tool for treating HF by using its pleiotropic effects. In addition to its use in patients with diabetes mellitus who are at high cardiovascular risk or who have already experienced a cardiovascular event, SGLT2i administration has been shown to have positive effects on a variety of HF manifestations and stages, regardless of the patient's presence of diabetes mellitus. Material and Methods: According to the guide, 110 patients (83 males and 27 females) with heart failure with mildly reduced ejection fraction (HFmrEF), with T2D and neoplasia, were enrolled in the prospective study. The structural and functional state of the left ventricle myocardium and ejection fraction was assessed through echocardiography. Patients were randomized to receive once-daily dapagliflozin 10 mg. Results: Patients with HFmrEF were divided into 3 subgroups according to age. 7% (8) patients aged < 45 years, 35% (28) patients aged between 46-59 years, and 58% (74) patients aged> 60 years. The most prevalent comorbidities were hypertension (43.1%), coronary heart disease (40%), and obesity (33.2%). Study drug discontinuation and serious adverse events were not frequent in the subgroups, in either men or women, until now. Conclusions: SGLT-2 inhibitors are a novel class of antidiabetic agents that have demonstrated positive efficacy and safety outcomes in the setting of HFmrEF. Until now, in our study, dapagliflozin was safe and well-tolerated irrespective of sex.

Keywords: diabetes mellitus type 2, Sodium-glucose co-transporters-2 inhibitors, heart failure, neoplasia

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