Search results for: interfacial activity

Authors: Vanja Misic, Mohamed El-Mogy, Yousef Haj-Ahmad

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Endonuclease G (EndoG) is a well conserved mitochondrio-nuclear nuclease with dual lethal and vital roles in the cell. The aim of our study was to examine whether EndoG exerts its nuclease activity on exogenous DNA substrates such as plasmid DNA (pDNA), considering their importance in gene therapy applications. The effects of EndoG knockdown on pDNA stability and levels of encoded reporter gene expression were evaluated in the cervical carcinoma HeLa cells. Transfection of pDNA vectors encoding short-hairpin RNAs (shRNAs) reduced levels of EndoG mRNA and nuclease activity in HeLa cells. In physiological circumstances, EndoG knockdown did not have an effect on the stability of pDNA or the levels of encoded transgene expression as measured over a four day time-course. However, when endogenous expression of EndoG was induced by an extrinsic stimulus, targeting of EndoG by shRNA improved the perceived stability and transgene expression of pDNA vectors. Therefore, EndoG is not a mediator of exogenous DNA clearance, but in non-physiological circumstances it may non-specifically cleave intracellular DNA regardless of its origin. These findings make it unlikely that targeting of EndoG is a viable strategy for improving the duration and level of transgene expression from non-viral DNA vectors in gene therapy efforts.

Keywords: EndoG, silencing, exogenous DNA stability, HeLa cells

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Authors: Katja Siefken, Astrid Junge

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Background: Research indicates that physical activity (PA) protects us from developing mental disorders. The knowledge regarding optimal domain, intensity, type, context, and amount of PA promotion for the prevention of mental disorders is sparse and incoherent. The objective of this study is to determine the relationship between PA domains and mental well-being, and whether associations vary by domain, amount, context, intensity, and type of PA. Methods: 310 individuals (age: 25 yrs., SD 7; 73% female) completed a questionnaire on personal patterns of their PA behaviour (IPQA) and their mental health (Centre of Epidemiologic Studies Depression Scale (CES-D), Generalized Anxiety Disorder (GAD-7) scale, the subjective physical well-being (FEW-16)). Linear and multiple regression were used for analysis. Findings: Individuals who met the PA recommendation (N=269) reported higher scores on subjective physical well-being than those who did not meet the PA recommendations (N=41). Whilst vigorous intensity PA predicts subjective well-being (β = .122, p = .028), it also correlates with depression. The more vigorously physically active a person is, the higher the depression score (β = .127, p = .026). The strongest impact of PA on mental well-being can be seen in the transport domain. A positive linear correlation on subjective physical well-being (β =.175, p = .002), and a negative linear correlation for anxiety (β =-.142, p = .011) and depression (β = -.164, p = .004) was found. Multiple regression analysis indicates similar results: Time spent in active transport on the bicycle significantly lowers anxiety and depression scores and enhances subjective physical well-being. The more time a participant spends using the bicycle for transport, the lower the depression (β = -.143, p = .013) and anxiety scores (β = -.111,p = .050). Conclusions: Meeting the PA recommendations enhances subjective physical well-being. Active transport has a substantial impact on mental well-being. Findings have implications for policymakers, employers, public health experts and civil society. A stronger focus on the promotion and protection of health through active transport is recommended. Inter-sectoral exchange, outside the health sector, is required. Health systems must engage other sectors in adopting policies that maximize possible health gains.

Keywords: active transport, mental well-being, health promotion, psychological disorders

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Authors: F. Haddadj, S. Hamdi, M. Khames, A. Kadi, S. Zenia, A. Smai, H. Saadi, B. Doumandji-Mitiche

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The use of a biopesticide based on a microorganism scale requires particular care including safety against the useful auxiliary fauna and mammals among other human beings. Due to its persistence in soil and its apparent human and animal safety, Beauveria bassiana is a cryptogram used for controlling pests organizations, particularly in the locust where its effectiveness has been proven by several highly studies. This fungus is also called for greater respect for biotic communities and the environment. Indeed, biopesticides have several environmental benefits: biodegradability, their activity and selectivity decrease unintended non-target species effects, decreased resistance to some of them. It is in this sense that we contribute by presenting our work on the safety of B. bassiana against mammals. For this we conducted a toxicological study of this fungus strain on Wistar rats Rattus norvegicus, first its effect on weight gain. In a second time were performed histological target organ is the liver. After 20 days of treatment, the results of the toxicological studies have shown that B. bassiana caused no change in the physiological state of rats or weight gain, behavior and diet. On cuts in liver histology revealed no disturbance on the organ.

Keywords: entomopathogenic fungus, B. bassiana, Rattus norvegicus, toxicological study, environment

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Authors: Amandeep Sharma, Digvaijay Verma, Ruplal Choudhary

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Food processing is an activity across the globe that help in better handling of agricultural produce, including dairy, meat, and fish. The operations carried out in the food industry includes raw material quality authenticity; sorting and grading; processing into various products using thermal treatments – heating, freezing, and chilling; packaging; and storage at the appropriate temperature to maximize the shelf life of the products. All this is done to safeguard the food products and to ensure the distribution up to the consumer. The approaches to develop predictive models based on mathematical or statistical tools or empirical models’ development has been reported for various milk processing activities, including plant maintenance and wastage. Recently AI is the key factor for the fourth industrial revolution. AI plays a vital role in the food industry, not only in quality and food security but also in different areas such as manufacturing, packaging, and cleaning. A new conceptual model was developed, which shows that smaller sample size as only spectra would be required to predict the other values hence leads to saving on raw materials and chemicals otherwise used for experimentation during the research and new product development activity. It would be a futuristic approach if these tools can be further clubbed with the mobile phones through some software development for their real time application in the field for quality check and traceability of the product.

Keywords: predictive modlleing, ann, ai, food

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Authors: Chiung-Hui Chen

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The Internet of Things (IoT) was designed for widespread convenience. With the smart tag and the sensing network, a large quantity of dynamic information is immediately presented in the IoT. Through the internal communication and interaction, meaningful objects provide real-time services for users. Therefore, the service with appropriate decision-making has become an essential issue. Based on the science of human behavior, this study employed the environment model to record the time sequences and locations of different behaviors and adopted the probability module of the hierarchical Hidden Markov Model for the inference. The statistical analysis was conducted to achieve the following objectives: First, define user behaviors and predict the user behavior routes with the environment model to analyze user purposes. Second, construct the hierarchical Hidden Markov Model according to the logic framework, and establish the sequential intensity among behaviors to get acquainted with the use and activity fabric of the intelligent environment. Third, establish the intensity of the relation between the probability of objects’ being used and the objects. The indicator can describe the possible limitations of the mechanism. As the process is recorded in the information of the system created in this study, these data can be reused to adjust the procedure of intelligent design services.

Keywords: behavior, big data, hierarchical hidden Markov model, intelligent object

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Authors: Mary Ghobrial, Sahar Wefky

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Marine macroalgae have proven to be rich source of bioactive compounds with biomedical potential, not only for human but also for veterinary medicine. Emergence of microbial disease in aquaculture industries implies serious loses. Usage of commercial antibiotics for fish disease treatment produces undesirable side effects. Marine organisms are a rich source of structurally novel biologically active metabolites. Competition for space and nutrients led to the evolution of antimicrobial defense strategies in the aquatic environment. The interest in marine organisms as a potential and promising source of pharmaceutical agents has increased in the last years. Many bioactive and pharmacologically active substances have been isolated from microalgae. Compounds with antibacterial, antifungal and antiviral activities have been also detected in green, brown and red algae. Selected species of marine benthic algae belonging to the Phaeophyta and Rhodophyta, collected from different coastal areas of Alexandria (Egypt), were investigated for their antibacterial and antifungal, activities. Macroalgae samples were collected during low tide from the Alexandria Mediterranean coast. Samples were air dried under shade at room temperature. The dry algae were ground, using electric mixer grinder. They were soaked in 10 ml of each of the solvents acetone, ethanol, methanol and hexane. Antimicrobial activity was evaluated using well-cut diffusion technique In vitro screening of organic solvent extracts from the marine macroalgae Laurencia pinnatifida, Pterocladia capillaceae, Stepopodium zonale, Halopteris scoparia and Sargassum hystrix, showed specific activity in inhibiting the growth of five virulent strains of bacteria pathogenic to fish Pseudomonas fluorescens, Aeromonas hydrophila, Vibrio anguillarum, V. tandara, Escherichia coli and two fungi Aspergillus flavus and A. niger. Results showed that, acetone and ethanol extracts of all test macroalgae exhibited antibacterial activity, while acetone extract of the brown Sargassum hystrix displayed the highest antifungal activity. The extracts of seaweeds inhibited bacteria more strongly than fungi and species of the Rhodophyta showed the greatest activity against the bacteria rather than fungi tested. The gas liquid chromatography coupled with mass spectrometry detection technique allows good qualitative and quantitative analysis of the fractionated extracts with high sensitivity to the smaller amounts of components. Results indicated that, the main common component in the acetone extracts of L. pinnatifida and P. capillacea is 4-hydroxy-4-methyl2-pentanone representing 64.38 and 58.60%. Thus, the extracts derived from the red macroalgae were more efficient than those obtained from the brown macroalgae in combating bacterial pathogens rather than pathogenic fungi. The most preferred species over all was the red Laurencia pinnatifida. In conclusion, the present study provides the potential of red and brown macroalgae extracts for development of anti-pathogenic agents for use in fish aquaculture.

Keywords: bacteria, fungi, extracts, solvents

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Authors: Dmitry Yu. Korulkin, Raissa A. Muzychkina, Evgenii N. Kojaev

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In review the generalized data about biological activity of anthraquinone-containing plants and specimens on their basis is presented. Data of traditional medicine, results of bioscreening and clinical researches of specimens are analyzed.

Keywords: anthraquinones, physiologically active substances, phytopreparation, Ramon

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Authors: S. Lekhavat, U. Srimongkoluk, P. Ratanachamnong, G. Laungsopapun

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Quercetin was encapsulated with whey protein and high methoxyl pectin by spray drying technique. Feed solution, consisting of 0.1875 0.125 and 0.0625 % w/w quercetin, respectively, was prepared and then sprays at outlet temperature of 70, 80 and 90 °C. Quercetin contents either in feed solution or in spray dried powder were determined by HPLC technique. Physicochemical properties such as viscosity and total soluble solid of feed solution as well as moisture content and water activity of spray dried powder were examined. Particle morphology was imaged using scanning electron microscope. The results showed that feed solution has total soluble solid and viscosity in range of 1.73-5.60 ºBrix and 2.58-8.15 cP, in that order. After spray drying, the moisture content and water activity value of powder are in range of 0.58-2.72 % and 0.18-0.31, respectively. Quercetin content in dried sample increased along with outlet drying temperature but decreased when total soluble solid increased. It was shown that particles are likely to shrivel when spray drying at high temperature. The suggested conditions for encapsulation of quercetin are feed solution with 0.0625 % (w/w) quercetin and spray drying at drying outlet temperature of 90°C.

Keywords: drying temperature, particle morphology, spray drying, quercetin

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Authors: Medhanie Gebremedhin Gebru, Alex Schechter

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Dimethyl ether (DME) possesses several advantages over other small organic molecules such as methanol, ethanol, and ammonia in terms of providing higher energy density, being less toxic, and having lower Nafion membrane crossover. However, the absence of an active and stable catalyst has been the bottleneck that hindered the commercialization of direct DME fuel cells. A Vulcan XC72 carbon-supported ternary metal catalyst, Pt₃Pd₃Sn₂/C is reported to have yielded the highest specific power density (90 mW mg-¹PGM) as compared to other catalysts tested fordirect DME fuel cell (DDMEFC). However, the micropores and sulfur groups present in Vulcan XC72 hinder the fuel utilization by causing Pt agglomeration and sulfur poisoning. Vulcan XC72 having a high carbon sp³ hybridization content, is also prone to corrosion. Therefore, carbon supports such as multi-walled carbon nanotube (MWCNT), black pearl 2000 (BP2000), and their cold N2 plasma-treated counterpartswere tested to further enhance the activity of the catalyst, and the outputs with these carbons were compared with the originally used support. Detailed characterization of the pristine and carbon supports was conducted. Electrochemical measurements in three-electrode cells and laboratory prototype fuel cells were conducted.Pt₃Pd₃Sn₂/BP2000 exhibited excellent performance in terms of electrochemical active surface area (ECSA), peak current density (jp), and DME oxidation charge (Qoxi). The effect of the plasma activation on the activity improvement was observed only in the case of MWCNT while having little or no effect on the other carbons. A Pt₃Pd₃Sn₂ supported on the optimized mixture of carbons containing 75% plasma-activated MWCNT and 25% BP2000 (Pt₃Pd₃Sn₂/75M25B) provided the highest reported power density of 117 mW mg-1PGM using an anode loading of1.55 mgPGMcm⁻².

Keywords: DME, DDMEFC, ternary metal catalyst, carbon support, plasma activation

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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: Indranil Chakraborty, Kalyan Mandal

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Three new α-hydroxy carboxylate group functionalized MnFe₂O₄ nanoparticles (NPs) have been developed to explore the microscopic origin of ligand modified fluorescence and magnetic properties of nearly monodispersed MnFe₂O₄ NPs. The surface functionalization has been carried out with three small organic ligands (tartrate, malate, and citrate) having different number of α-hydroxy carboxylate functional group along with steric effect. Detailed study unveils that α-hydroxy carboxylate moiety of the ligands plays key role to generate intrinsic fluorescence in functionalized MnFe₂O₄ NPs through the activation of ligand to metal charge transfer transitions, associated with ligand-Mn²⁺/Fe³⁺ interactions along with d-d transition corresponding to d-orbital energy level splitting of Fe³⁺ ions on NP surface. Further, MnFe₂O₄ NPs show a maximum 140.88% increase in coercivity and 97.95% decrease in magnetization compared to its bare one upon functionalization. The ligands that induce smallest crystal field splitting of d-orbital energy level of transition metal ions are found to result in strongest ferromagnetic activation of the NPs. Finally, our developed tartrate functionalized MnFe₂O₄ (T-MnFe₂O₄) NPs have been utilized for studying DNA binding interaction and nuclease activity for stimulating their beneficial activities toward diverse biomedical applications. The spectroscopic measurements indicate that T-MnFe₂O₄ NPs bind calf thymus DNA by intercalative mode. The ability of T-MnFe₂O₄ NPs to induce DNA cleavage was studied by gel electrophoresis technique where the complex is found to promote the cleavage of pBR322 plasmid DNA from the super coiled form I to linear coiled form II and nicked coiled form III with good efficiency. This may be taken into account for designing new biomolecular detection agents and anti-cancer drug which can open up a new door toward diverse non-invasive biomedical applications.

Keywords: MnFe₂O₄ nanoparticle, α-hydroxy carboxylic acid, comparative fluorescence, magnetism study, DNA interaction, nuclease activity

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Authors: Khalid Ahmed Elrabie Abdelrasoul

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The present study was carried out on Sahiwal cattle bulls maintained at the Artificial Breeding Complex, NDRI, Karnal, Hayana, India, to assess the effect of cooling using mist cooling and fanning on Sahiwal bulls in the dry hot summer season. Fourteen Sahiwal bulls were divided into two groups of seven each. Sexual behavior and semen quality traits considered were: Reaction time (RT), Dismounting time (DMT), Total time taken in mounts (TTTM), Flehmen response (FR), Erection Score (ES), Protrusion Score (PS), Intensity of thrust (ITS), Temperament Score (TS), Libido Score (LS), Semen volume, Physical appearance, Mass activity, Initial progressive motility, Non-eosinophilic spermatozoa count (NESC) and post thaw motility percent. Data were analyzed by least squares technique. Group-1 was the control, whereas group-2 (treatment group) bulls were exposed to mist cooling and fanning (thrice a day 15 min each) in the dry hot summer season. Group-2 showed significantly (p < 0.01) higher value in DMT (sec), ES, PS, ITS, LS, semen volume (ml), semen color density, mass activity, initial motility, progressive motility and live sperm.

Keywords: mist cooling, Sahiwal bulls, semen quality, sexual behavior

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Authors: Maria P. Geneva, Ira V. Stancheva, Marieta G. Hristozkova, Roumiana D. Vasilevska-Ivanova, Mariana T. Sichanova, Janet R. Mincheva

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Hyssopus officinalis L., Lamiaceae, commonly called hyssop, is an aromatic, semi-evergreen, woody-based, shrubby perennial plant. Hyssop is a good expectorant and antiviral herb commonly used to treat respiratory conditions such as influenza, sinus infections, colds, and bronchitis. Most of its medicinal properties are attributed to the essential oil of hyssop. The study was conducted to evaluate the influence of inoculation with arbuscular mycorrhizal fungi of in vitro propagated hyssop plants on the: activities of antioxidant enzymes superoxide dismutase, catalase, guaiacol peroxidase and ascorbate peroxidase; accumulation of non-enzymatic antioxidants total phenols and flavonoid, water-soluble soluble antioxidant metabolites expressed as ascorbic acid; the antioxidant potential of hyssop methanol extracts assessed by two common methods: free radical scavenging activity using free stable radical (2,2-diphenyl-1-picrylhydrazyl, DPPH• and ferric reducing antioxidant power FRAP in flowers and leaves. The successfully adapted to field conditions in vitro plants (survival rate 95%) were inoculated with arbuscular mycorrhizal fungi (Claroideoglomus claroideum, ref. EEZ 54). It was established that the activities of enzymes with antioxidant capacity (superoxide dismutase, catalase, guaiacol peroxidase and ascorbate peroxidase) were significantly higher in leaves than in flowers in both control and mycorrhized plants. In flowers and leaves of inoculated plants, the antioxidant enzymes activity were lower than in non-inoculated plants, only in SOD activity, there was no difference. The content of low molecular metabolites with antioxidant capacity as total phenols, total flavonoids, and water soluble antioxidants was higher in inoculated plants. There were no significant differences between control and inoculated plants both for FRAP and DPPH antioxidant activity. According to plant essential oil content, there was no difference between non-inoculated and inoculated plants. Based on our results we could suggest that antioxidant capacity of in vitro propagated hyssop plant under conditions of cultivation is determined by the phenolic compounds-total phenols and flavonoids as well as by the levels of water-soluble metabolites with antioxidant potential. Acknowledgments: This study was conducted with financial support from National Science Fund at the Bulgarian Ministry of Education and Science, Project DN06/7 17.12.16.

Keywords: antioxidant enzymes, antioxidant metabolites, arbuscular mycorrhizal fungi, Hyssopus officinalis L.

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Authors: Velvizhi Dharmalingam, Rajalaksmi Ramalingam, Rekha Prabhu, Ilavarasan Raju

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Background: The use of herbal products for various therapeutic regimens has increased tremendously in the developing countries. Elaeocarpus ganitrus(Rudraksha) is a broad-leaved tree, belonging to the family Elaeocarpaceae found in tropical and subtropical areas. It is popular in an indigenous system of medicine like Ayurveda, Siddha, and Unani. According to Ayurvedic medicine, Rudraksha is used in the managing of blood pressure, asthma, mental disorders, diabetes, gynaecological disorders, neurological disorders such as epilepsy and liver diseases. Objectives: The present study aimed to study the physicochemical parameters of Elaeocarpus ganitrus(fruits) and identify the phenolic compounds (gallic acid, ellagic acid, and chebulinic acid). To estimate the microbial load and the antibacterial activity of extract of Elaeocarpus ganitrus for selective pathogens. Methodology: The dried powdered fruit of Elaeocarpus ganitrus was performed the physicochemical parameters (such as Loss on drying, Alcohol soluble extractive, Water soluble extractive, Total ash and Acid insoluble ash) and pH was measured. The dried coarse powdered fruit of Elaeocarpus ganitrus was extracted successively with hexane, chloroform, ethylacetate and aqueous alcohol by cold percolation method. Identification of phenolic compounds (gallic acid, ellagic acid, chebulinic acid) was done by HPTLC method and confirmed by co-TLC using different solvent system.The successive extracts of Elaeocarpus ganitrus and standards (like gallic acid, ellagic acid, and chebulinic acid) was approximately weighed and made up with alcohol. HPTLC (CAMAG) analysis was performed on a TLC over silica gel 60F254 precoated aluminium plate, layer thickness 0.2 mm (E.Merck, Germany) by using ATS4, Visualizer and Scanner with wavelength at 254 nm, 366 nm and derivatized with different reagents. The microbial load such as total bacterial count, total fungal count, Enterobacteria, Escherichia coli, Salmonella species, Staphylococcus aureus and Pseudomonas aeruginosa by serial dilution method and antibacterial activity of was measured by Kirby bauer method for selective pathogens. Results: The physicochemical parameter of Elaeocarpus ganitrus was studied for standardization of crude drug. Among all the successive extracts were identified with phenolic compounds and Elaeocarpus ganitrus extract having potent antibacterial activity against gram-positive and gram-negative bacteria.

Keywords: antimicrobial activity, Elaeocarpus ganitrus, HPTLC, phenolic compounds

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Authors: A. M. Gadanya, M. S. Sule

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Effect of oral administration of Gadagi tea on some antioxidant enzymes was assessed in healthy male albino rats. The rats were grouped and administered with standard doses of the 3 types of Gadagi tea i.e. Sak, Sada and Magani for a period of four weeks. Animals that were not administered with the tea constituted the control group. At the end of fourth week, the animals were sacrificed and their serum superoxide dismutase (SOD), glutathione reductase (GR) and catalase (CAT) activities were determined. The activities of the enzymes were also determined in the brain, liver, kidney and intestine homogenates of the rats. Mean SOD activity in brain of rats orally administered with “sada” was found to be significantly higher (P < 0.05) than that of the control group. Mean CAT activity in the intestine of rats orally administered with “magani” was found to be significantly higher (P < 0.05) than that of the control group and the experimental groups of Sak and Sada at standard dose level. Thus, all the “Gadagi” tea preparations studied at standard dose level could stimulate antioxidant enzymes, especially SOD in brain and CAT in intestine (by Sada) and CAT in intestine (by Magani).

Keywords: “Gadagi” tea, superoxide dismutase, catalase, glutathione reductase

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Authors: Rafat M. Mohareb

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

Keywords: estrone, heterocyclization, cytotoxicity, biomedicine

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Authors: Valentina Obradović, Josip Mesić, Maja Ergović Ravančić, Kamila Mijowska, Brankica Svitlica

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French oak and American oak barrels are most famous all over the world, but barrels of different origin can also be used for obtaining high quality wines. The aim of this research was to compare the influence of different Slavonian (Croatian) and French oak barrels on the quality of Chardonnay wine. Grapes were grown in Croatian wine growing region of Kutjevo in 2015. Chardonnay wines were tested for basic oenological parameters (alcohol, extract, reducing sugar, SO2, acidity), total polyphenols content (Folin-Ciocalteu method), antioxidant activity (ABTS and DPPH method) and color density. Sensory evaluation was performed by students of viticulture/oenology. Samples produced by classical fermentation and ageing in French oak barrels, had better results for polyphenols and sensory evaluation (especially low toasting level) than samples in Slavonian barrels. All tested samples were scored as a “quality” or “premium quality” wines. Sur lie method of fermentation and ageing in Slavonian oak barrel had very good extraction of polyphenols and high antioxidant activity with the usage of authentic yeasts, while commercial yeast strain resulted in worse chemical and sensory parameters.

Keywords: chardonnay, French oak, Slavonian oak, sur lie

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Authors: Sahriye Sonmez, Sedat Citak

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

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

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Authors: Samar Saadeldin Abdelmotalab Omer, Ikram Mohamed Eltayeb Elsiddig, Saad Mohammed Hussein Ayoub

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A variety of herbal medicinal plants are known to confer beneficial effects in regards to modification of cardiovascular ri’=sk factors. The anti-hypercholesterolaemic and antioxidant activities of the crude ethanolic extracts of Citrus reticulate fruit peel, Zingiber officinale rhizome and Sesamum indicum seed extracts have been demonstrated. These plants are assumed to possess biologically active principles, which impart their pharmacologic activities. GC-MS analysis of the ethanolic extracts was carried out to identify the active principles and their percentages of occurrence in the analytes. Analysis of the extracts was carried out using (GS-MS QP) type Schimadzu 2010 equipped with a capillary column RTX-50 (restec), (length 30mm, diameter 0.25mm, and thickness 0.25mm). Helium was used as a carrier gas, the temperature was programmed at 200°C for 5 minutes at a rate of 15ml/minute, and the extracts were injected using split injection mode. The identification of different components was achieved from their Mass Spectra and Retention time, compared with those in the NIST library. The results revealed the presence of 80 compounds in Sudanese locally grown C. reticulata fruit peel extract, most of which were monoterpenoid compounds including Limonene (3.03%), Alpha & Gamma - terpinenes (2.61%), Linalool (1.38%), Citral (1.72%) which are known to have profound antioxidant effects. The Sesquiterpenoids Humulene (0.26%) and Caryophyllene (1.97%) were also identified, the latter known to have profound anti-anxiety and anti-depressant activity in addition to the beneficiary effects in lipid regulation. The analysis of the locally grown S. indicum oily and water soluble portions of seed extract revealed the presence of a total of 64 compounds with considerably high percentage of the mono-unsaturated fatty acid ester methyl oleate (66.99%) in addition to methyl stearate (9.35%) and palmitate (15.71%) of oil portion, whereas, plant sterols including Gamma-sitosterol (13.5%), fucosterol (2.11%) and stigmasterol (1.95%) in addition to gamma-tocopherol (1.16%) were detected in extract water-soluble portion. The latter indicate various principles known to have valuable pharmacological benefits including antioxidant activities and beneficiary effects on intestinal cholesterol absorption and regulation of serum cholesterol levels. Z. officinale rhizome extract analysis revealed the presence of 93 compounds, the most abundant were alpha-zingeberine (16.5%), gingerol (9.25%), alpha-sesquiphellandrene (8.3%), zingerone (6.78%), beta-bisabolene (4.19%), alpha-farnesene (3.56%), ar-curcumene (3.29%), gamma-elemene (1.25%) and a variety of other compounds. The presence of these active principles reflected on the activity of the extract. Activity could be assigned to a single or a combination of two or more extract components. GC-MS analysis concluded the occurrence of compounds known to possess antioxidant activity and lipid profile effects.

Keywords: gas chromatography, indicum, officinale, reticulata

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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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Authors: Ghavamaldin Asadian, Aptin Rahnavard, Mariamalsadat Taghavi

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Achillea millefolium L. is one of the most important medicinal plants with antioxidant compounds. The use of compounds derived from plants reduces the incidence of many chronic diseases. The purpose of this investigation is study of total phenolic content and antioxidant activity some of ecotypes yarrow grown in natural habitats in Iran. This experimental study was conducted in 2013 at the Islamic Azad University, Tonekabon Branch. After identifying the natural sites, we have attempted to harvest of aerial part and after drying in lab temperature, essential oil was extracted by steam distillation. In this research for evaluate the antioxidant properties was used of three method, DPPH, Antioxidant capacity ferro revival and phosphomolybdenum, that all mechanism is based on the electron donating. All ecotypes had antioxidant activity and ecotypes grown in Kandovan region were measured with the most total phenolic (89.5 mg GA/g dew) and flavonoid (20.4 µg/g dew) and the lowest in Saveh (71.3 mg GA/g dew, 17.4 µg/g dew). Variation of the antioxidant properties were significant (P≤0.01) in areas and were accounted Kandovan with highest value and the lowest in Save. As a result, yarrow essential oil grown in Kandovan in terms of amount of total phenolic, flavonoid and antioxidant property, it was determined the best natural habitat.

Keywords: achillea millefolium L., antioxidant compounds, DPPH, total phenolic, flavonoid natural habitats

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Authors: N. W. I. A. Jayawardana, W. A. T. A. Jayalath, W. M. T. Madhujith, U. Ralapanawa, R. S. Jayasekera, S. A. S. B. Alagiyawanna, A. M. K. R. Bandara, N. S. Kalupahana

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There is mounting evidence to the effect that dietary and lifestyle changes affect the incidence of non-communicable diseases (NCDs). This study was conducted to investigate the association of diet, physical activity, smoking, alcohol consumption and duration of sleep with overweight, obesity, hypertension and diabetes in a cohort of males from the Central Province of Sri Lanka. A total of 2694 individuals aged between 17 – 68 years (Mean = 31) were included in the study. Body Mass Index cutoff values for Asians were used to categorize the participants as normal, overweight and obese. The dietary data were collected using a food frequency questionnaire [FFQ] and data on the level of physical activity, smoking, alcohol consumption and sleeping hours were obtained using a self-administered validated questionnaire. Systolic and diastolic blood pressure, random blood glucose levels were measured to determine the incidence of hypertension and diabetes. Among the individuals, the prevalence of overweight and obesity were 34% and 16.4% respectively. Approximately 37% of the participants suffered from hypertension. Overweight and obesity were associated with older age men (P<0.0001), frequency of smoking (P=0.0434), alcohol consumption level (P=0.0287) and the quantity of lipid intake (P=0.0081). Consumption of fish (P=0.6983) and salty snacks (P=0.8327), sleeping hours (P=0.6847) and the level of physical activity were not significantly (P=0.3301) associated with the incidence of overweight and obesity. Based on the fitted model, only age was significantly associated with hypertension (P < 0.001). Further, age (P < 0.0001), sleeping hours (P=0.0953) and consumption of fatty foods (P=0.0930) were significantly associated with diabetes. Age was associated with higher odds of pre diabetes (OR:1.089;95% CI:1.053,1.127) and diabetes (OR:1.077;95% CI:1.055,1.1) whereas 7-8 hrs. of sleep per day was associated with lesser odds of diabetes (OR:0.403;95% CI:0.184,0.884). High prevalence of overweight, obesity and hypertension in working-age males is a threatening sign for this area. As this population ages in the future and urbanization continues, the prevalence of above risk factors will likely to escalate.

Keywords: age, males, non-communicable diseases, obesity

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Authors: Ksenija Milošević, Davor Lončarević, Tihana Mudrinić, Jasmina Dostanić

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Heterogeneous photocatalytic processes have gained growing interest as an efficient method to generate hydrogen by using clean energy sources and degrading various organic pollutants. The main obstacles that restrict efficient photoactivity are narrow light-response range and high rates of charge carrier recombination. The formation of heterojunction by combining a semiconductor with low VB and a semiconductor with high CB and a suitable band gap was found to be an efficient method to prepare more sensible materials with improved charge separation, appropriate oxidation and reduction ability, and enhanced visible-light harvesting. In our research, various binary heterojunction systems based on the wide-band gap (TiO₂) and narrow bandgap (g-C₃N₄, CuO, and Co₂O₃) photocatalyst were studied. The morphology, optical, and electrochemical properties of the photocatalysts were analyzed by X-ray diffraction (XRD), scanning electron microscopy (FE-SEM), N₂ physisorption, diffuse reflectance measurements (DRS), and Mott-Schottky analysis. The photocatalytic performance of the synthesized catalysts was tested in single and simultaneous systems. The synthesized photocatalysts displayed good adsorption capacity and enhanced visible-light photocatalytic performance. The mutual interactions of pollutants on their adsorption and degradation efficiency were investigated. The interfacial connection between photocatalyst constituents and the mechanism of the transport pathway of photogenerated charge species was discussed. A radical scavenger study revealed the interaction mechanisms of the photocatalyst constituents in single and multiple pollutant systems under solar and visible light irradiation, indicating the type of heterojunction system (Z scheme or type II).

Keywords: bandgap alignment, heterojunction, photocatalysis, reaction mechanism

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Authors: Shabnam Sarwar, Franck Lacoeuille, Nadia Withofs, Roland Hustinx

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After the development of a potential surrogate of MAA, and its successful application for the diagnosis of pulmonary embolism in artificially embolized rats’ lungs, this microparticulate system were radiolabelled with gallium-68 to synthesize 68Ga-SBMP with high radiochemical purity >99%. As a prerequisite step of clinical trials, 68Ga- labelled starch based microparticles (SBMP) were analysed for their in-vivo behavior in small animals. The purpose of the presented work includes the ex-vivo biodistribution studies of 68Ga-SBMP in order to assess the activity uptake in target organs with respect to time, excretion pathways of the radiopharmaceutical, %ID/g in major organs, T/NT ratios, in-vivo stability of the radiotracer and subsequently the microparticles in the target organs. Radiolabelling of starch based microparticles was performed by incubating it with 68Ga generator eluate (430±26 MBq) at room temperature and pressure without using any harsh reaction condition. For Ex-vivo biodistribution studies healthy White Wistar rats weighing between 345-460 g were injected intravenously 68Ga-SBMP 20±8 MBq, containing about 2,00,000-6,00,000 SBMP particles in a volume of 700µL. The rats were euthanized at predefined time intervals (5min, 30min, 60min and 120min) and their organ parts were cut, washed, and put in the pre-weighed tubes and measured for radioactivity counts through automatic Gamma counter. The 68Ga-SBMP produced >99% RCP just after 10-20 min incubation through a simple and robust procedure. Biodistribution of 68Ga-SBMP showed that initially just after 5 min post injection major uptake was observed in the lungs following by blood, heart, liver, kidneys, bladder, urine, spleen, stomach, small intestine, colon, skin and skeleton, thymus and at last the smallest activity was found in brain. Radioactivity counts stayed stable in lungs with gradual decrease with the passage of time, and after 2h post injection, almost half of the activity were seen in lungs. This is a sufficient time to perform PET/CT lungs scanning in humans while activity in the liver, spleen, gut and urinary system decreased with time. The results showed that urinary system is the excretion pathways instead of hepatobiliary excretion. There was a high value of T/NT ratios which suggest fine tune images for PET/CT lung perfusion studies henceforth further pre-clinical studies and then clinical trials should be planned in order to utilize this potential lung perfusion agent.

Keywords: starch based microparticles, gallium-68, biodistribution, target organs, excretion pathways

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Authors: Mirjana Petronijević, Sanja Panić, Aleksandra Cvetanović, Branko Kordić, Nenad Grba

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Enzyme peroxidases are biological catalysts and play a major role in phenolic wastewater treatments and other environmental applications. The most studied species from the peroxidases family is horseradish peroxidase (HRP). In environmental processes, HRP could be used in its free or immobilized form. Enzyme immobilization onto solid support is performed to improve the enzyme properties, prolong its lifespan and operational stability and allow its reuse in industrial applications. One of the enzyme supports of a newer generation is magnetic particles (MPs). Fe₃O₄ MPs are the most widely pursued immobilization of enzymes owing to their remarkable advantages of biocompatibility and non-toxicity. Also, MPs can be easily separated and recovered from the water by applying an external magnetic field. On the other hand, metals and metal oxides are not suitable for the covalent binding of enzymes, so it is necessary to perform their surface modification. Fe₃O₄ MPs functionalization could be performed during the process of their synthesis if it takes place in the presence of plant extracts. Extracts of plant material, such as wild plants, herbs, even waste materials of the food and agricultural industry (bark, shell, leaves, peel), are rich in various bioactive components such as polyphenols, flavonoids, sugars, etc. When the synthesis of magnetite is performed in the presence of plant extracts, bioactive components are incorporated into the surface of the magnetite, thereby affecting its functionalization. In this paper, the suitability of bio-magnetite as solid support for covalent immobilization of HRP across glutaraldehyde was examined. The activity of immobilized HRP at different pH values (4-9) and temperatures (20-80°C) and reusability were examined. Bio-MP was synthesized by co-precipitation method from Fe(II) and Fe(III) sulfate salts in the presence of water extract of the Allium cepa peel. The water extract showed 81% of antiradical potential (according to DPPH assay), which is connected with the high content of polyphenols. According to the FTIR analysis, the bio-magnetite contains oxygen functional groups (-OH, -COOH, C=O) suitable for binding to glutaraldehyde, after which the enzyme is covalently immobilized. The immobilized enzyme showed high activity at ambient temperature and pH 7 (30 U/g) and retained ≥ 80% of its activity at a wide range of pH (5-8) and temperature (20-50°C). The HRP immobilized onto bio-MPs showed remarkable stability towards temperature and pH variations compared to the free enzyme form. On the other hand, immobilized HRP showed low reusability after the first washing cycle enzyme retains 50% of its activity, while after the third washing cycle retains only 22%.

Keywords: bio-magnetite, enzyme immobilization, water extracts, environmental protection

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Authors: Arosha Rashmi De Silva, Ananda Chandrasekara

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This study seeks to comprehensively examine the influence of fast-food consumption and physical activity levels on the body weight of young employees within the private sector of Sri Lanka. The escalating popularity of fast food has raised concerns about its nutritional content and associated health ramifications. To investigate this phenomenon, a cohort of 100 individuals aged between 25 and 45, employed in Sri Lanka's private sector, participated in this research. These participants provided socio-demographic data through a standardized questionnaire, enabling the characterization of their backgrounds. Additionally, participants disclosed their frequency of fast-food consumption and engagement in physical activities, utilizing validated assessment tools. The collected data was meticulously compiled into an Excel spreadsheet and subjected to rigorous statistical analysis. Descriptive statistics, such as percentages and proportions, were employed to delineate the body weight status of the participants. Employing chi-square tests, our study identified significant associations between fast-food consumption, levels of physical activity, and body weight categories. Furthermore, through binary logistic regression analysis, potential risk factors contributing to overweight and obesity within the young employee cohort were elucidated. Our findings revealed a disconcerting trend, with 6% of participants classified as underweight, 32% within the normal weight range, and a substantial 62% categorized as overweight or obese. These outcomes underscore the alarming prevalence of overweight and obesity among young private-sector employees, particularly within the bustling urban landscape of Colombo, Sri Lanka. The data strongly imply a robust correlation between fast-food consumption, sedentary behaviors, and higher body weight categories, reflective of the evolving lifestyle patterns associated with the nation's economic growth. This study emphasizes the urgent need for effective interventions to counter the detrimental effects of fast-food consumption. The implementation of awareness campaigns elucidating the adverse health consequences of fast food, coupled with comprehensive nutritional education, can empower individuals to make informed dietary choices. Workplace interventions, including the provision of healthier meal alternatives and the facilitation of physical activity opportunities, are essential in fostering a healthier workforce and mitigating the escalating burden of overweight and obesity in Sri Lanka

Keywords: fast food consumption, obese, overweight, physical activity level

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Authors: Babak Sadeghi, Parisa Ghayomipour

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ZnO/Ag nanocomposites coated with polyvinyl chloride (PVC) were prepared by chemical reduction method, for anti-infection biomaterial application. There is a growing interest in attempts in using biomolecular as the templates to grow inorganic nanocomposites in controlled morphology and structure. By optimizing the experiment conditions, we successfully fabricated high yield of ZnO/Ag nanocomposite with full coverage of high-density polyvinyl chloride (PVC) coating. More importantly, ZnO/Ag nanocomposites were shown to significantly inhibit the growth of S. aureus in solution. It was further shown that ZnO/Ag nanocomposites induced thiol depletion that caused death of S. aureus. The coatings were fully characterized using techniques such as scanning electron microscopy (SEM), transmission electron microscopy (TEM) and X-ray diffraction (XRD). Most importantly, compared to uncoated metals, the coatings on PVC promoted healthy antibacterial activity. Importantly, compared to ZnO-Ag -uncoated PVC, the ZnO/Ag nanocomposites coated was approximately triplet more effective in preventing bacteria attachment. The result of Thermal Gravimetric Analysis (TGA) indicates that, the ZnO/Ag nanocomposites are chemically stable in the temperature range from 50 to 900 ºC. This result, for the first time, demonstrates the potential of using ZnO/Ag nanocomposites as a coating material for numerous anti-bacterial applications.

Keywords: nanocomposites, antibacterial activity, scanning electron microscopy (SEM), x-ray diffraction (XRD)

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Authors: Belal Rahhal, Isra Taha, Insaf Najajreh, Waleed Basha, Hamzeh Alzabadeh, Ahed Zyoud

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Phytochemical Investigation and Diuretic Activity of the Palestinian Crataegus aronia in Mice using an Aqueous Extract Division of Physiology, Pharmacology and Toxicology Faculty of Medicine and Health Sciences An- Najah National University Nablus- Palestine Belal Rahhal, Isra Taha, Insaf Najajreh, Waleed Basha, Hamzeh Alzabadeh and Ahed Zyoud Purpose: Throughout history, various natural materials were used as remedies for treatment of various diseases, and recently a vastly growing and renewed interest in herbal medicine is witnessed globally. In Palestinian folk medicine, Crataegus aronia is used as a diuretic and for treatment of hypertension. This study aimed to assess the preliminary phytochemical properties and the diuretic effect of the aqueous extracts of this plant in mice after its intraperitonial administration. Methods: It is an experimental trial applied on mice (n=8, Male, CD-1, weight range: [25-30 gram]), which are divided into two groups (4 in each). The first group administered with the plant extract (500 mg/kg) , and the second with normal saline as negative control group. Then urine output and electrolyte contents were quantified up to 6 hours for the three groups and then compared to the control one. Results: Preliminary phytochemical screening reveals the presence of tannins, alkaloids and flavoniods as major phytoconstituents in aqueous extract. Significant diuresis was noted in those received the aqueous extract of Crataegus aronia (p < 0.05) compared to controls. Moreover, aqueous extract had an acidic pH and a mild increase in the electrolyte excretion (Na, K). Conclusions: Our results revealed that Crataegus aronia aqueous extract has a potential diuretic effect. Further studies are needed to evaluate this diuretic effect in the relief of diseases characterized by volume overload. Keywords: C. aronia, furosemide, diuresis, mice, medicinal plants.

Keywords: medicinal plants, diuretic activity, mice, C. aronia, , furosemide, , Phytochemical Investigation

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Authors: Vera Lukasova, Eva Filova, Jana Dankova, Vera Sovkova, Matej Daniel, Michala Rampichova

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Proper bone implants should promote fast adhesion of cells, stimulate cell differentiation and support the formation of bone tissue. Nowadays titanium is used as a biocompatible material capable of bone tissue integration. This study was focused on comparison of bioactive properties of two titanium alloys - beta titanium alloy Ti36Nb6Ta and standard medical titanium alloy Ti6A14V. The advantage of beta titanium alloy Ti36Nb6Ta is mainly that this material does not contain adverse elements like vanadium or aluminium. Titanium alloys were sterilized in ethanol, placed into 48 well plates and seeded with porcine mesenchymal stem cells. Cells were cultivated for 14 days in standard growth cultivation media with osteogenic supplements. Cell metabolic activity was quantified using MTS assay (Promega). Cell adhesion on day 1 and cell proliferation on further days were verified immunohistochemically using beta-actin monoclonal antibody and secondary antibody conjugated with AlexaFluor®488. Differentiation of cells was evaluated using alkaline phosphatase assay. Additionally, gene expression of collagen I was measured by qRT-PCR. Porcine mesenchymal stem cells adhered and spread well on beta titanium alloy Ti36Nb6Ta on day 1. During the 14 days’ time period the cells were spread confluently on the surface of the beta titanium alloy Ti36Nb6Ta. The metabolic activity of cells increased during the whole cultivation period. In comparison to standard medical titanium alloy Ti6A14V, we did not observe any differences. Moreover, the expression of collagen I gene revealed no statistical differences between both titanium alloys. Therefore, a beta titanium alloy Ti36Nb6Ta promotes cell adhesion, metabolic activity, proliferation and collagen I expression equally to standard medical titanium alloy Ti6A14V. Thus, beta titanium is a suitable material that provides sufficient biocompatible properties. This project was supported by the Czech Science Foundation: grant No. 16-14758S.

Keywords: beta titanium alloy, biocompatibility, differentiation, mesenchymal stem cells

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Authors: Pintusorn Hansakul, Ruchilak Rattarom, Arunporn Itharat

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Background: Benjakul, a Thai traditional herbal formulation, comprises of five plants: Piper chaba, Piper sarmentosum, Piper interruptum, Plumbago indica, and Zingiber officinale. It has been widely used to treat cancer patients in the context of folk medicine in Thailand. This study aimed to investigate the cytotoxic effect of the ethanol extract of Benjakul against three non-small cell lung cancer (NSCLC) cell lines (NCI-H226, A549, COR-L23), small cell lung cancer (SCLC) cell line NCI-H1688 and normal lung fibroblast cell line MRC-5. The study further examined the molecular mechanisms underlying its cytotoxicity via induction of apoptosis in NCI-H226 cells. Methods: The cytotoxic effect of Benjakul was determined by SRB assay. The effect of Benjakul on cell cycle distribution was assessed by flow cytometric analysis. The apoptotic effects of Benjakul were determined by sub-G1 quantitation and Annexin V-FITC/PI flow cytometric analyses as well as by changes in caspase-3 activity. Results: Benjakul exerted potent cytotoxicity on NCI-H226 and A549 cells but lower cytotoxicity on COR-L23 and NCI-H1688 cells without any cytotoxic effect on normal cells. Molecular studies showed that Benjakul extract induced G2/M phase arrest in human NCI-H226 cells in a dose-dependent manner. The highest concentration of Benjakul (150 μg/ml) led to the highest increase in the G2/M population at 12 h, followed by the highest increase in the sub-G1 population (apoptotic cells) at 60 h. Benjakul extract also induced early apoptosis (AnnexinV +/PI−) in NCI-H226 cells in a dose- and time- dependent manner. Moreover, treatment with 150 μg/ml Benjakul extract for 36 h markedly increased caspase-3 activity by 3.5-fold, and pretreatment with the general caspase inhibitor z-VAD-fmk completely abolished such activity. Conclusions: This study reveals for the first time the regulation of apoptosis in human lung cancer NCI-H226 cells through caspase-dependent mechanism by Benjakul extract.

Keywords: apoptosis, Benjakul, caspase activation, cytotoxicity

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