Search results for: In vitro

Authors: A. N. Gornostaeva, P. I. Bobyleva, E. R. Andreeva, L. B. Buravkova

Abstract:

Multipotent mesenchymal stromal cells (MSCs) possess immunomodulatory properties. The effect of MSCs on the crucial cellular immunity compartment – T-cells is of a special interest. It is known that MSC tissue niche and expected milieu of their interaction with T- cells are characterized by low oxygen concentration, whereas the in vitro experiments usually are carried out at a much higher ambient oxygen (20%). We firstly evaluated immunomodulatory effects of MSCs on T-cells at tissue-related oxygen (5%) after interaction implied cell-to-cell contacts and paracrine factors only. It turned out that MSCs under reduced oxygen can effectively suppress the activation and proliferation of PHAstimulated T-cells and can provoke decrease in the production of proinflammatory and increase in anti-inflammatory cytokines. In hypoxia some effects were amplified (inhibition of proliferation, antiinflammatory cytokine profile shift). This impact was more evident after direct cell-to-cell interaction; lack of intercellular contacts could revoke the potentiating effect of hypoxia.

Keywords: Cell-to-cell interaction, low oxygen, MSC immunosuppression, T-cells.

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Authors: C.K. Hindumathy

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Alcohol and water extracts of Cymbopogon citratus was investigated for anti-bacterial properties and phytochemical constituents. The extract was screened against four gram-negative bacteria Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Proteus vulgaris) and two grampositive bacteria Bacillus subtilis and Staphylococcus aureus at four different concentrations (1:1, 1:5, 1:10 and 1:20) using disc diffusion method. The antibacterial examination was by disc diffusion techniques, while the photochemical constituents were investigated using standard chemical methods. Results showed that the extracts inhibited the growth of standard and local strains of the organisms used. The treatments were significantly different (P = 0.05). The minimum inhibitory concentration of the extracts against the tested microorganisms ranged between 150mg/ml and 50mg/ml. The alcohol extracts were found to be generally more effective than the water extract. The photochemical analysis revealed the presence of alkaloids and phenol but absence of cardiac and cyanogenic glycosides. The presence of alkaloid and phenols were inferred as being responsible for the anti-bacterial properties of the extracts.

Keywords: Cymbopogon citratus; gram negative and grampositive

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Authors: Chun-Yen Sung, Chung-Yao Yang, Tzu-Chun Liao, Wen-Shiang Chen, Chao-Min Cheng, J. Andrew Yeh

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The study describes chitosan membrane platform modified with nanostructure pattern which using nanotechnology to fabricate. The cell-substrate interaction between neuro-2a neuroblasts cell lines and chitosan membrane (flat, nanostructure and nanostructure pattern types) was investigated. The adhered morphology of neuro-2a cells depends on the topography of chitosan surface. We have found that neuro-2a showed different morphogenesis when cells adhered on flat and nanostructure chitosan membrane. The cell projected area of neuro-2a on flat chitosan membrane is larger than on nanostructure chitosan membrane. In addition, neuro-2a cells preferred to adhere on flat chitosan surface region than on nanostructure chitosan membrane to immobilize and differentiation. The experiment suggests surface topography can be used as a critical mechanism to isolate group of neuro-2a to a particular rectangle area on chitosan membrane. Our finding will provide a platform to take patch clamp to record electrophysiological behavior about neurons in vitro in the future.

Keywords: Chitosan membrane, neuro-2a, wet chemical etching, solvent casting.

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Authors: Masaki Yamaguchi, Akira Ito, Noriaki Okamoto, Yoshinori Kawabe, Masamichi Kamihira

Abstract:

Heat-inducible gene expression vectors are useful for hyperthermia-induced cancer gene therapy, because the combination of hyperthermia and gene therapy can considerably improve the therapeutic effects. In the present study, we developed an enhanced heat-inducible transgene expression system in which a heat-shock protein (HSP) promoter and tetracycline-responsive transactivator were combined. When the transactivator plasmid containing the tetracycline-responsive transactivator gene was co-transfected with the reporter gene expression plasmid, a high level of heat-induced gene expression was observed compared with that using the HSP promoter without the transactivator. In vitro evaluation of the therapeutic effect using HeLa cells showed that heat-induced therapeutic gene expression caused cell death in a high percentage of these cells, indicating that this strategy is promising for cancer gene therapy.

Keywords: Inducible gene expression, Gene therapy, Hyperthermia, Heat shock protein, Tetracycline transactivator.

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Authors: M Tayari, SZ Moosavi-nejad, A Shabani, M Rezaei Tavirani

Abstract:

Free Hemoglobin promotes the accumulation of hydroxyl radicals by the heme iron, which can react with endogenous hydrogen peroxide to produce free radicals which may cause severe oxidative cell damage. Haptoglobin binds to Hemoglobin strongly and Haptoglobin-Hemoglobin binding is irreversible. Peroxidase activity of Haptoglobin(2-2)-Hemoglobin complex was assayed by following increase of absorption of produced tetraguaiacol as the second substrate of Haptoglobin-Hemoglobin complex at 470 nm and 42°C by UV-Vis spectrophotometer. The results have shown that peroxidase activity of Haptoglobin(2-2)-Hemoglobin complex is modulated via homotropic effect of hydrogen peroxide as allostric substrate. On the other hand antioxidant property of Haptoglobin(2- 2)-Hemoglobin was increased via heterotropic effect of the two drugs (especially ampicillin) on peroxidase activity of the complex. Both drugs also have mild effect on quality of homotropic property of peroxidase activity of Haptoglobin(2-2)-Hemoglobin complex. Therefore, in vitro studies show that the two drugs may help Hp-Hb complex to remove hydrogen peroxide from serum at pathologic temperature ature (42 C).

Keywords: Haptoglobin, Hemoglobin, Antioxidant, Antibiotics.

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Authors: Irina Spaka, Rita Birkenfelde, Svetlana Kozireva, Jevgenija Osmjana, Madara Upmane, ElenaKashuba, Irina Kholodnyuk Holodnuka

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Epstein-Barr virus (EBV) is implicated in the pathogenesis of the endemic Burkitt-s lymphoma (BL). The EBVpositive BL-derived cell lines initially maintain the original tumor phenotype of EBV infection (latency I, LatI), but most of them drift toward a lymphoblast phenotype of EBV latency III (LatIII) during in vitro culturing. The aim of the present work was to characterize the B-cell subsets in EBV-positive BL cell lines and to verify whether a particular cell subset correlates with the type of EBV infection. The phenotype analysis of two EBV-negative and eleven EBV-positive (three of LatI and eight of LatIII) BL cell lines was performed by polychromatic flow cytomery, based on expression pattern of CD19, CD10, CD38, CD27, and CD5 markers. Two cell subsets, CD19+CD10+ and CD19+CD10-, were defined in LatIII BL cell lines. In both subsets, the CD27 and CD5 cell surface expression was detected in a proportion of the cells.

Keywords: B-cell subsets, Burkitt's lymphoma cell lines, EBV latency, phenotype profiles.

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Authors: Marine A. Balayan, Susanna S. Mirzabekyan, Marine Isajanyan, Zaven S. Pepoyan, Аrmen H. Trchounian, Аstghik Z. Pepoyan, Helena Bujdakova

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It has been shown that pH 7,3 and 37 0C are the optimal condition for the growth of E. coli “ASAP". The cells grow well on Glucose, Lactose, D-Mannitol, D-Sorbitol, (+)-Xylose, L- (+)-Arabinose and Dulcitol. No growth has been observed on Sucrose, Inositol, Phenylalanine, and Tryptophan. The strain is sensitive to a range of antibiotics. The present study has demonstrated that E. coli “ASAP" inhibit the growth of S. enterica ATCC #700931 in vitro. The studies on conjugating activity has revealed no conjugant of E. coli “ASAP" with plasmid strains E. coli G35#59 and S. enterica ATCC #700931. On the other hand, the conjugants with low frequencies were obtained from E. coli “ASAP" with E. coli G35#61, and E. coli “ASAP" with randomly chosen isolate from healthy human gut microflora: E. coli E6. The results of present study have demonstrated improvements in gut microflora condition of patients with different diseases after the administration of “ASAP"

Keywords: E. coli, ASAP, Probiotic formula, gut microflora.

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Authors: Librado A. Santiago, Anna Beatriz R. Mayor

Abstract:

Alongside with antioxidant, pro-oxidant activity is also observed in phytochemical compounds. In the study, Ficus odorata, an endemic medicinal plant in the Philippines, was screened for the potential medical application of its pro-oxidant activity.

Phytochemical screening revealed the presence of terpenes, glycosides and phenolic acids. The crude extract was found to contain low gallic acid and quercetin equivalence. The TLC chromatogram of the crude extract showed that none of the 11 spots obtained has antioxidant activity nor correspond to gallic acid and quercetin standards. Experiments showed that the crude extract has stimulatory activity towards DPPH radicals, hydrogen peroxide, hydroxyl radicals, superoxide anions and nitric oxide. Moreover, the extract exhibited a low ferric reducing power.

The prooxidant activity was evident in the crude ethanolic leaf extract of F. odorata, which may provide a better understanding of the plant’s pharmacological importance in the prevention of diseases.

Keywords: Ficus odorata Blanco, Free Radicals, Oxidative Stress, Prooxidant, Antioxidant.

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Authors: Majid Mirzaei, Saeid Samiezadeh , Abbas Khodadadi, Mohammad R. Ghazavi

Abstract:

This paper presents a novel method for prediction of the mechanical behavior of proximal femur using the general framework of the quantitative computed tomography (QCT)-based finite element Analysis (FEA). A systematic imaging and modeling procedure was developed for reliable correspondence between the QCT-based FEA and the in-vitro mechanical testing. A speciallydesigned holding frame was used to define and maintain a unique geometrical reference system during the analysis and testing. The QCT images were directly converted into voxel-based 3D finite element models for linear and nonlinear analyses. The equivalent plastic strain and the strain energy density measures were used to identify the critical elements and predict the failure patterns. The samples were destructively tested using a specially-designed gripping fixture (with five degrees of freedom) mounted within a universal mechanical testing machine. Very good agreements were found between the experimental and the predicted failure patterns and the associated load levels.

Keywords: Bone, Osteoporosis, Noninvasive methods, Failure Analysis

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Authors: H. N. Mehmanparast, J.M. Mac-Thiong., Y. Petit

Abstract:

Transpedicular screw fixation in spinal fractures, degenerative changes, or deformities is a well-established procedure. However, important rate of fixation failure due to screw bending, loosening, or pullout are still reported particularly in weak bone stock in osteoporosis. To overcome the problem, mechanism of failure has to be fully investigated in vitro. Post-mortem human subjects are less accessible and animal cadavers comprise limitations due to different geometry and mechanical properties. Therefore, the development of a synthetic model mimicking the realistic human vertebra is highly demanded. A bone surrogate, composed of Polyurethane (PU) foam analogous to cancellous bone porous structure, was tested for 3 different densities in this study. The mechanical properties were investigated under uniaxial compression test by minimizing the end artifacts on specimens. The results indicated that PU foam of 0.32 g.cm-3 density has comparable mechanical properties to human cancellous bone in terms of young-s modulus and yield strength. Therefore, the obtained information can be considered as primary step for developing a realistic cancellous bone of human vertebral body. Further evaluations are also recommended for other density groups.

Keywords: Cancellous bone, Pedicle screw, Polyurethane foam, Synthetic bone

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Authors: Y.K. Kim, Y.S. Jang, H.H. Park, J.H. Ji, I.S. Park, T.S. Bae, M.H. Lee

Abstract:

Magnesium is used implant material potentially for non-toxicity to the human body. Due to the excellent bio-compatibility, Mg alloys is applied to implants avoiding removal second surgery. However, it is found commercial magnesium alloys including aluminum has low corrosion resistance, resulting subcutaneous gas bubbles and consequently the approach as permanent bio-materials. Generally, Aluminum is known to pollution substance, and it raises toxicity to nervous system. Therefore especially Mg-35Zn-3Ca alloy is prepared for new biodegradable materials in this study. And the pulsed power is used in constant-current mode of DC power kinds of anodization. Based on the aforementioned study, it examines corrosion resistance and biocompatibility by effect of current and frequency variation. The surface properties and thickness were compared using scanning electronic microscopy. Corrosion resistance was assessed via potentiodynamic polarization and the effect of oxide layer on the body was assessed cell viability. Anodized Mg-35Zn-3Ca alloy has good biocompatibility in vitro by current and frequency variation.

Keywords: Biodegradable material, Mg, anodization, osteoblast cell, pulse power.

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Authors: May-Ling Tan, Yi Su, Chi-Wan Lim, Liang Zhong, Ru-San Tan

Abstract:

This paper describes an automatic algorithm to restore the shape of three-dimensional (3D) left ventricle (LV) models created from magnetic resonance imaging (MRI) data using a geometry-driven optimization approach. Our basic premise is to restore the LV shape such that the LV epicardial surface is smooth after the restoration. A geometrical measure known as the Minimum Principle Curvature (κ2) is used to assess the smoothness of the LV. This measure is used to construct the objective function of a two-step optimization process. The objective of the optimization is to achieve a smooth epicardial shape by iterative in-plane translation of the MRI slices. Quantitatively, this yields a minimum sum in terms of the magnitude of κ 2, when κ2 is negative. A limited memory quasi-Newton algorithm, L-BFGS-B, is used to solve the optimization problem. We tested our algorithm on an in vitro theoretical LV model and 10 in vivo patient-specific models which contain significant motion artifacts. The results show that our method is able to automatically restore the shape of LV models back to smoothness without altering the general shape of the model. The magnitudes of in-plane translations are also consistent with existing registration techniques and experimental findings.

Keywords: Magnetic Resonance Imaging, Left Ventricle, ShapeRestoration, Principle Curvature, Optimization

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Authors: Münüre Tanur Erkoyuncu, Mustafa Yorgancılar

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Crop improvement through genetic engineering depends on effective and reproducible plant regeneration systems. Immature embryos are the most widely used explant source for in vitro regeneration in barley (Hordeum vulgare L.). However, immature embryos require the continuous growth of donor plants and the suitable stage for their culture is also certainly limited. On the other hand, mature embryos can be procured and stored easily; they can be studied throughout the year. In this study, an effective callus induction and plant regeneration were aimed to develop from mature embryos of different barley genotypes. The effect of medium (MS₁ and MS₂), auxin type (2,4-D, dicamba, picloram and 2,4,5-T) and concentrations (2, 4, 6 mg/l) on callus formation and effect of cytokinin type (TDZ, BAP) and concentrations (0.2, 0.5, 1.0 mg/l) on green plant regeneration were evaluated in mature embryo culture of barley. Callus and shoot formation was successful for all genotypes. By depending on genotype, MS₁ is the best medium, 4 mg/l dicamba is the best growth regulator in the callus induction and MS₁ is the best medium, 1 mg/l BAP is the best growth regulator in the shoot formation were determined.

Keywords: Barley, callus, embryo culture, mature embryo.

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Authors: YuriyTasinkevych, Ihor Trots, AndrzejNowicki, Marcin Lewandowski

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The paper presents the optimization problem for the multi-element synthetic transmit aperture method (MSTA) in ultrasound imaging applications. The optimal choice of the transmit aperture size is performed as a trade-off between the lateral resolution, penetration depth and the frame rate. Results of the analysis obtained by a developed optimization algorithm are presented. Maximum penetration depth and the best lateral resolution at given depths are chosen as the optimization criteria. The optimization algorithm was tested using synthetic aperture data of point reflectors simulated by Filed II program for Matlab® for the case of 5MHz 128-element linear transducer array with 0.48 mm pitch are presented. The visualization of experimentally obtained synthetic aperture data of a tissue mimicking phantom and in vitro measurements of the beef liver are also shown. The data were obtained using the SonixTOUCH Research systemequipped with a linear 4MHz 128 element transducerwith 0.3 mm element pitch, 0.28 mm element width and 70% fractional bandwidth was excited by one sine cycle pulse burst of transducer's center frequency.

Keywords: synthetic aperture method, ultrasound imaging, beamforming.

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Authors: Brou K., Guéhi T., Konan A. G., Gbakayoro J. B., Gnakri D.

Abstract:

Two types of crushing were applied to grains of red sorghum: manual crushing using a mortar and pestle of kitchen and mechanical crushing using a hammer mill. The flours obtained at the end of these various crushing were filtered and subdivided in different fractions according to the diameters of the mesh of the sieves (0.16mm; 0.25mm; 0.315mm; 0.4mm, and 0.63mm…). Some physical, chemical and nutritional traits of these flours were evaluated using Association of Official Analytical Chemists (AOAC). In vitro digestibility of these flours was also studied with freezing of flour 1% like substrate and α-amylase from B. licheniformis (E.C.3.2.1.1; Megazyme, Wicklow, Ireland). The results revealed that the batches of flours which have the finest diameters as 0.16mm; 0.25mm are the richest one in nutrients and are also the most digestible. Also mechanical crushing is the best mean to obtain significant amount of flours. In conclusion, the type of crushing and the size of the particles have an impact on the final concentration of some nutrients of the flours obtained. Indeed, the finest particles (0.16mm – 0.25mm 0.315mm) obtained after sifting of the flours are more nutritive and have a better digestibility than others size. So the finest particles could be advised for management of cereals namely the sorghum for the production of the infantile foods.

Keywords: Nutrients, digestibility, crush, flour, milling, granulometry.

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Authors: Andreas Heindl, Alexander K. Seewald, Martin Schepelmann, Radu Rogojanu, Giovanna Bises, Theresia Thalhammer, Isabella Ellinger

Abstract:

Bone remodeling occurs by the balanced action of bone resorbing osteoclasts (OC) and bone-building osteoblasts. Increased bone resorption by excessive OC activity contributes to malignant and non-malignant diseases including osteoporosis. To study OC differentiation and function, OC formed in in vitro cultures are currently counted manually, a tedious procedure which is prone to inter-observer differences. Aiming for an automated OC-quantification system, classification of OC and precursor cells was done on fluorescence microscope images based on the distinct appearance of fluorescent nuclei. Following ellipse fitting to nuclei, a combination of eight features enabled clustering of OC and precursor cell nuclei. After evaluating different machine-learning techniques, LOGREG achieved 74% correctly classified OC and precursor cell nuclei, outperforming human experts (best expert: 55%). In combination with the automated detection of total cell areas, this system allows to measure various cell parameters and most importantly to quantify proteins involved in osteoclastogenesis.

Keywords: osteoclasts, machine learning, ellipse fitting.

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Authors: M. E. Abalaka, S. Y. Daniyan, S. O. Adeyemo, D. Damisa

Abstract:

Gold nanoparticles (AuNPs) have gained increasing interest in recent times. This is greatly due to their special features, which include unusual optical and electronic properties, high stability and biological compatibility, controllable morphology and size dispersion, and easy surface functionalization. In typical synthesis, AuNPs were produced by reduction of gold salt AuCl4 in an appropriate solvent. A stabilizing agent was added to prevent the particles from aggregating. The antibacterial activity of different sizes of gold nanoparticles was investigated against Staphylococcus aureus, Salmonella typhi and Pseudomonas pneumonia using the disk diffusion method in a Müeller–Hinton Agar. The Au-NPs were effective against all bacteria tested. That the Au-NPs were successfully synthesized in suspension and were used to study the antibacterial activity of the two medicinal plants against some bacterial pathogens suggests that Au-NPs can be employed as an effective bacteria inhibitor and may be an effective tool in medical field. The study clearly showed that the Au-NPs exhibiting inhibition towards the tested pathogenic bacteria in vitro could have the same effects in vivo and thus may be useful in the medical field if well researched into.

Keywords: Gold Nanoparticles, Gomphrena celesioides, Prunus amygdalus, Pathogens.

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Authors: Nagasamy Venkatesh Dhandapani, Amged Awad El-Gied

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Cefixime, a BCS class II drug, is insoluble in water but freely soluble in acetone and in alcohol. The aqueous solubility of cefixime in water is poor and exhibits exceptionally slow and intrinsic dissolution rate. In the present study, cefixime and β-Cyclodextrin (β-CD) solid dispersions were prepared with a view to study the effect and influence of β-CD on the solubility and dissolution rate of this poorly aqueous soluble drug. Phase solubility profile revealed that the solubility of cefixime was increased in the presence of β-CD and was classified as A_L-type. Effect of variable, such as drug:carrier ratio, was studied. Physical characterization of the solid dispersion was characterized by Fourier transform infrared spectroscopy (FT-IR) and Differential scanning calorimetry (DSC). These studies revealed that a distinct loss of drug crystallinity in the solid molecular dispersions is ostensibly accounting for enhancement of dissolution rate in distilled water. The drug release from the prepared solid dispersion exhibited a first order kinetics. Solid dispersions of cefixime showed a 6.77 times fold increase in dissolution rate over the pure drug.

Keywords: Cefixime, β-Cyclodextrin, solid dispersions, kneading method, dissolution, release kinetics.

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Authors: B. Dikici, I. Ozdemir, M. Topuz

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The corrosion behaviour of 316L stainless steel coatings obtained by cold spray method was investigated in this study. 316L powders were deposited onto Al5052 aluminum substrates. The coatings were produced using nitrogen (N₂) process gas. In order to further improve the corrosion and mechanical properties of the coatings, heat treatment was applied at 250 and 750 °C. The corrosion performances of the coatings were compared using the potentiodynamic scanning (PDS) technique under in-vitro conditions (in Ringer’s solution at 37 °C). In addition, the hardness and porosity tests were carried out on the coatings. Microstructural characterization of the coatings was carried out by using scanning electron microscopy attached with energy dispersive spectrometer (SEM-EDS) and X-ray diffraction (XRD) technique. It was found that clean surfaces and a good adhesion were achieved for particle/substrate bonding. The heat treatment process provided both elimination of the anisotropy in the coating and resulting in healing-up of the incomplete interfaces between the deposited particles. It was found that the corrosion potential of the annealed coatings at 750 °C was higher than that of commercially 316 L stainless steel. Moreover, the microstructural investigations after the corrosion tests revealed that corrosion preferentially starts at inter-splat boundaries.

Keywords: 316L, biomaterials, cold spray, heat treatment.

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Authors: Dehpour A. A., Eslami B., Rezaie S., Hashemian S. F., Shafie F., Kiaie M.

Abstract:

The aims of study were investigation on chemical composition essential oil and the effect of extract of Coronilla varia on antimicrobial and cytotoxicity activity. The essential oils of Coronilla varia is obtained by hydrodistillation and analyzed by (GC/MS) for determining their chemical composition and identification of their components. Antibacterial activity of plant extract was determined by disc diffusion method and anticancer activity measured by MTT assay. The major components in essential oil were Caryophyllene Oxide (60.19%), Alphacadinol (4.13%) and Homoadantaneca Robexylic Acid (3.31%). The extracts from Coronilla varia had interesting activity against Proteus mirabilis in the concentration of 700 μg/disc and did not show any activity against Staphylococus aureus, Bacillus subtillis, Klebsiella pneumonia and Entrobacter cloacae. The positive control, Ampicillin, Chloramphenicol and Cenphalothin had shown zone of inhibition resistant all bacteria. The ethanol extract of Corohilla varia inhibited on MCF7 cell lines. IC50 0.6(mg/ml) was the optimum concentration of extract from Coronilla varia inhibition of cell line growth. The MCF7 cancer cell line and Proteus mirabilis were more sensitive to Coronilla varia ethanol extract.

Keywords: Coronilla varia, Essential oil, Antibacterial, Anticancer, HeLa cell line.

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Authors: Kittiwann Nimkerdphol, Masahiro Nakagawa

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Biological reactions of individuals of a testing animal to toxic substance are unique and can be used as an indication of the existing of toxic substance. However, to distinguish such phenomenon need a very complicate system and even more complicate to analyze data in 3 dimensional. In this paper, a system to evaluate in vitro biological activities to acute toxicity of stochastic self-affine non-stationary signal of 3D goldfish swimming by using fractal analysis is introduced. Regular digital camcorders are utilized by proposed algorithm 3DCCPC to effectively capture and construct 3D movements of the fish. A Critical Exponent Method (CEM) has been adopted as a fractal estimator. The hypothesis was that the swimming of goldfish to acute toxic would show the fractal property which related to the toxic concentration. The experimental results supported the hypothesis by showing that the swimming of goldfish under the different toxic concentration has fractal properties. It also shows that the fractal dimension of the swimming related to the pH value of FD Ôëê 0.26pH + 0.05. With the proposed system, the fish is allowed to swim freely in all direction to react to the toxic. In addition, the trajectories are precisely evaluated by fractal analysis with critical exponent method and hence the results exhibit with much higher degree of confidence.

Keywords: 3D locomotion, bioassay, critical exponent method, CEM, fractal analysis, goldfish.

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Authors: Funmilola O. Omoya

Abstract:

Muscid flies are known to be vectors of disease agents and species that annoy humans and domesticated animals. An example of these flies is Musca domestica (house fly) whose adult and immature stages occur in a variety of filthy organic substances including household garbage and animal manures. They contribute to microbial contamination of foods. It is therefore imperative to control these flies as a result of their role in Public health. The second and third instars of Musca domestica (Linn) were infected with varying cell loads of Bacillus subtilis in vitro for a period of 48 hours to evaluate its larvicidal activities. Mortality of the larvae increased with incubation period after treatment with the varying cell loads. Investigation revealed that the second instars larvae were more susceptible to treatment than the third instars treatments. Values obtained from the third instar group were significantly different (P<0.05) from those obtained from the second instars group in all the treatments. Lethal concentration (LC₅₀) at 24 hours for 2^nd instars was 2.35 while LC₅₀ at 48 hours was 4.31.This study revealed that Bacillus subtilis possess good larvicidal potential for use in the control of Musca domestica in poultry farms.

Keywords: Bacillus subtilis, larvicidal activities, Musca domestica, poultry farms.

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Authors: Seyed Mohammad Hosseini, Amirhossein Moghanian

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The main purpose of this research was to design a biomimetic system by freeze-drying method for evaluating the effect of adding 5 and 10 mol. % of zinc (Zn) in 58S bioactive glass and gelatin (5ZnBG/G and 10ZnBG/G) in terms of structural and biological changes. The structural analyses of samples were performed by X-Ray Diffraction (XRD), scanning electron microscopy (SEM) and Fourier-transform infrared (FTIR) spectroscopy. Also, 3-(4,5dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and alkaline phosphatase (ALP) activity tests were carried out for investigation of MC3T3-E1 cell behaviors. The SEM results demonstrated the spherical shape of the formed hydroxyapatite (HA) phases and also HA characteristic peaks were detected by XRD spectroscopy after 3 days of immersion in the simulated body fluid (SBF) solution. Meanwhile, FTIR spectra proved that the intensity of P–O peaks for 5ZnBG/G was more than 10ZnBG/G and control samples. Moreover, the results of ALP activity test illustrated that the optimal amount of Zn (5ZnBG/G) caused a considerable enhancement in bone cell growth. Taken together, the scaffold with 5 mol.% Zn was introduced as an optimal sample because of its higher biocompatibility, in vitro bioactivity and growth of MC3T3-E1 cells in comparison with other samples in bone tissue engineering.

Keywords: Scaffold, gelatin, modified bioactive glass, ALP, bone tissue engineering.

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Authors: Ghada Salem, Nihal Kabel

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Objective: This in vitro study was carried out to assess the microleakage properties of nano-filled glass ionomer in comparison to resin-reinforced glass ionomers. Material and Methods: 40 deciduous molar teeth were included in this study. Class-II cavity was prepared in a standard form for all the specimens. The teeth were randomly distributed into two groups (20 per group) according to the restorative material used either nano-glass ionomer or Photac Fill glass ionomer restoration. All specimens were thermocycled for 1000 cycles between 5 and 55 °C. After that, the teeth were immersed in 2% methylene blue dye then sectioned and evaluated under a stereomicroscope. Microleakage was assessed using linear dye penetration and on a scale from zero to five. Results: Two way ANOVA test revealed a statistically significant lower degree of microleakage in both occlusal and gingival restorations (0.4±0.2), (0.9±0.1) for nano-filled glass ionomer group in comparison to resin modified glass ionomer (2.3±0.7), (2.4±0.5). No statistical difference was found between gingival and occlusal leakage regarding the effect of the measured site. Conclusion: Nano-filled glass ionomer shows superior sealing ability which enables this type of restoration to be used in minimum invasive treatment.

Keywords: Microleakage, nano-ionomer, resin-reinforced glass ionomer, proximal cavity preparation.

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Authors: Iman A. Elkiweri, Ph.D, Martha C. Tissot van Patot, Ph.D., Yan Ling Zhang, Ph.D., Uwe Christians, Ph.D., Thomas K. Henthorn, M.D.,

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Verapamil has been shown to inhibit fentanyl uptake in vitro and is a potent P-glycoprotein inhibitor. Tissue partitioning of loperamide, a commercially available opioid, is closely controlled by the P-gp efflux transporter. The following studies were designed to evaluate the effect of opioids on verapamil partitioning in the lung and brain, in vivo. Opioid (fentanyl or loperamide) was administered by intravenous infusion to Sprague Dawley rats alone or in combination with verapamil and plasma, with lung and brain tissues were collected at 1, 5, 6, 8, 10 and 60 minutes. Drug dispositions were modeled by recirculatory pharmacokinetic models. Fentanyl slightly increased the verapamil lung (PL) partition coefficient yet decreased the brain (PB) partition coefficient. Furthermore, loperamide significantly increased PLand PB. Fentanyl reduced the verapamil volume of distribution (V1) and verapamil elimination clearance (ClE). Fentanyl decreased verapamil brain partitioning, yet increased verapamil lung partitioning. Also, loperamide increased lung and brain partitioning in vivo. These results suggest that verapamil and fentanyl may be substrates of an unidentified inward transporter in brain tissue and confirm that verapamil and loperamide are substrates of the efflux transporter P-gp.

Keywords: Efflux transporter, elimination clearance, partition coefficient, verapamil

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Authors: S. Techaoei, K. Jarmkom, P. Eakwaropas, W. Khobjai

Abstract:

The objective of this research was focused on investigating in vitro antimicrobial activity of Phellinus linteus fruiting body extracts on Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus. Phellinus linteus fruiting body was extracted with ethanol and ethyl acetate and was vaporized. The disc diffusion assay was used to assess antimicrobial activity against tested bacterial strains. Primary screening of chemical profile of crude extract was determined by using thin layer chromatography. The positive control and the negative control were used as erythromycin and dimethyl sulfoxide, respectively. Initial screening of Phellinus linteus crude extract with the disc diffusion assay demonstrated that only ethanol had greater antimicrobial activity against Pseudomonas aeruginosa, Escherichia coli, Staphylococcus aureus and Methicillin-resistant Staphylococcus aureus. The MIC assay showed that the lower MIC was observed with 0.5 mg/ml of Pseudomonas aeruginosa and Methicillin-resistant Staphylococcus aureus and 0.25 mg/ml. of Escherichia coli and Staphylococcus aureus, respectively. TLC chemical profile of extract was represented at R_f ≈ 0.71-0.76.

Keywords: Staphylococcus aureus, Phellinus linteus, methicillin-resistant Staphylococcus aureus, antimicrobial activity, Escherichia coli.

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Authors: P. Thongmuang, Y. Sudjaroen

Abstract:

The methanolic extracts from seeds of tamarind (Tamarindus indica) was prepared by Soxhlet apparatus extraction and evaluated for total phenolic content by Folin-Ciocalteu method. Then, methanolic extract was screened biological activities (In vitro) for anti-melanogenic activity by tyrosinase inhibition test, antiinflammation activity by cyclooxygenase 1 (COX-1) and cyclooxygenase 2 (COX-2) inhibition test, and cytotoxic screening test with Vero cells. The results showed that total phenolic content, which contained in extract, was contained 27.72 mg of gallic acid equivalent per g of dry weight. The ability to inhibit tyrosinase enzyme, which exerted by Tamarind seed extracts (1 mg/ml) was 52.13 ± 0.42 %. The extract was not possessed inhibitory effect to COX-1 and COX-2 enzymes and cytotoxic effect to Vero cells. The finding is concludes that tested seed extract was possessed antimelanogenic activity with non-toxic effects. However, there was not exhibited anti-inflammatory activity. Further studies include the use of advance biological models to confirm this biological activity, as well as, the isolation and characterization of the purified compounds that it was contained.

Keywords: Tamarindus indica, anti-melanogenic, antiinflammatotion, cytotoxicity, seed, phenolic compounds.

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Authors: N. Cetinkaya, Y. S. Kuleyin

Abstract:

The aim of this study was to estimate the digestibility of the fruit internal skin of different varieties of hazelnuts to propose hazelnut fruit skin as an alternative feed source as roughage in ruminant nutrition. In 2015, the fruit internal skins of three different varieties of round hazelnuts (RH), pointed hazelnuts (PH) and almond hazelnuts (AH) were obtained from hazelnut processing factory then their crude nutrients analysis were carried out. Organic matter digestibility (OMD) and metabolisable energy (ME) values of hazelnut fruit skins were estimated from gas measured by in vitro gas production method. Their antioxidant activities were determined by spectrophotometric method. Crude nutrient values of three different varieties were; organic matter (OM): 87.83, 87.81 and 87.78%), crude protein (CP): 5.97, 5.93 and 5.89%, neutral detergent fiber (NDF): 30.30, 30.29 and 30.29%, acid detergent fiber (ADF): 48.68, 48.67 and 48.66% and acid detergent lignin (ADL): 25.43, 25.43 and 25.39% respectively. OMD from 24 h incubation time of RH, PH and AH were 22.04, 22.46 and 22.74%; ME_GP values were 3.69, 3.75 and 3.79 MJ/kg DM; and antioxidant activity values were 94.60, 94.54 and 94.52 IC 50 mg/mL respectively. The fruit internal skin of different varieties of hazelnuts may be considered as an alternative roughage for ruminant nutrition regarding to their crude and digestible nutritive values. Moreover, hazelnut fruit skin has a rich antioxidant content so it may be used as a feed additive for both ruminant and non-ruminant animals.

Keywords: Antioxidant activity, hazelnut fruit skin, metabolizable energy, organic matter digestibility.

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Authors: Chung-Yao Yang, Lin-Ya Huang, Tang-Long Shen, J. Andrew Yeh

Abstract:

The importance for manipulating an incorporated scaffold and directing cell behaviors is well appreciated for tissue engineering. Here, we developed newly nano-topographic oxidized silicon nanosponges capable of being various chemical modifications to provide much insight into the fundamental biology of how cells interact with their surrounding environment in vitro. A wet etching technique is exerted to allow us fabricated the silicon nanosponges in a high-throughput manner. Furthermore, various organo-silane chemicals enabled self-assembled on the surfaces by vapor deposition. We have found that Chinese hamster ovary (CHO) cells displayed certain distinguishable morphogenesis, adherent responses, and biochemical properties while cultured on these chemical modified nano-topographic structures in compared with the planar oxidized silicon counterparts, indicating that cell behaviors can be influenced by certain physical characteristic derived from nano-topography in addition to the hydrophobicity of contact surfaces crucial for cell adhesion and spreading. Of particular, there were predominant nano-actin punches and slender protrusions formed while cells were cultured on the nano-topographic structures. This study shed potential applications of these nano-topographic biomaterials for controlling cell development in tissue engineering or basic cell biology research.

Keywords: Nanosponge, Cell adhesion, Cell morphology

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Authors: Mithilesh Singh, Rakhi Chaturvedi

Abstract:

Neem is a highly heterozygous and commercially important perennial plant. Conventionally, it is propagated by seeds which loose viability within two weeks. Strictly cross pollinating nature of the plant causes serious barrier to the genetic improvement by conventional methods. Alternative methods of tree improvement such as somatic hybridization, mutagenesis and genetic transformation require an efficient in vitro plant regeneration system. In this regard, somatic embryogenesis particularly secondary somatic embryogenesis may offer an effective system for large scale plant propagation without affecting the clonal fidelity of the regenerants. It can be used for synthetic seed production, which further bolsters conservation of this tree species which is otherwise very difficult The present report describes the culture conditions necessary to induce and maintain repetitive somatic embryogenesis, for the first time, in neem. Out of various treatments tested, the somatic embryos were induced directly from immature zygotic embryos of neem on MS + TDZ (0.1 μM) + ABA (4 μM), in more than 76 % cultures. Direct secondary somatic embryogenesis occurred from primary somatic embryos on MS + IAA (5 μM) + GA3 (5 μM) in 12.5 % cultures. Embryogenic competence of the explant as well as of the primary embryos was maintained for a long period by repeated subcultures at frequent intervals. A maximum of 10 % of these somatic embryos were converted into plantlets.

Keywords: Azadirachta indica A. Juss., Cytokinin, Somatic embryogenesis, zygotic embryo culture.

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