Comparative Analysis of Chemical Composition and Biological Activities of Ajuga genevensis L. in in vitro Culture and Intact Plants

One of the tasks in contemporary biotechnology, pharmacology and other fields of human activities is to obtain biologically active substances from plants. They are very essential in the treatment of many diseases due to their actually high therapeutic value without visible side effects. However, sometimes the possibility of obtaining the metabolites is limited due to the reduction of wild-growing plants. That is why the plant cell cultures are of great interest as alternative sources of biologically active substances. Besides, during the monitored cultivation, it is possible to obtain substances that are not synthesized by plants in nature. Isolated culture of Ajuga genevensis with high growth activity and ability of regeneration was obtained using MS nutrient medium. The agar-diffusion method showed that aqueous extracts of callus culture revealed high antimicrobial activity towards various gram-positive (Bacillus subtilis A1WT; B. mesentericus WDCM 1873; Staphylococcus aureus WDCM 5233; Staph. citreus WT) and gram-negative (Escherichia coli WKPM M-17; Salmonella typhimurium TA 100) microorganisms. The broth dilution method revealed that the minimal and half maximal inhibitory concentration values against E. coli corresponded to the 70 μg/mL and 140 μg/mL concentration of the extract respectively. According to the photochemiluminescent analysis, callus tissue extracts of leaf and root origin showed higher antioxidant activity than the same quantity of A. genevensis intact plant extract. A. genevensis intact plant and callus culture extracts showed no cytotoxic effect on K-562 suspension cell line of human chronic myeloid leukemia. The GC-MS analysis showed deep differences between the qualitative and quantitative composition of callus culture and intact plant extracts. Hexacosane (11.17%); n-hexadecanoic acid (9.33%); and 2-methoxy-4-vinylphenol (4.28%) were the main components of intact plant extracts. 10-Methylnonadecane (57.0%); methoxyacetic acid, 2-tetradecyl ester (17.75%) and 1-Bromopentadecane (14.55%) were the main components of A. genevensis callus culture extracts. Obtained data indicate that callus culture of A. genevensis can be used as an alternative source of biologically active substances.

Anti-Inflammatory Activity of Topical Anthocyanins by Complexation and Niosomal Encapsulation

Anthocyanins are natural pigments with effective UV protection but their topical use could be limited due to their physicochemical characteristics. An attempt to overcome such limitations by complexation of 2 major anthocyanin-rich sources, C. ternatea and Z. mays, has potentiated its use as topical antiinflammatory. Cell studies indicate no cytotoxicity of the anthocyanin complex (AC) up to 1 mg/ml tested in HaCaT and human fore head fibroblasts by MTT. Croton oil-induced ear edema in Wistar rats suggests an effective dose of 5 mg/cm2 of AC as a topical anti-inflammatory in comparison to 0.5 mg/cm2 of fluocinolone acetonide. Niosomal encapsulation of the AC significantly prolonged the anti-inflammatory activity particularly at 8 h after topical application (p = 0.0001). The AC was not cytotoxic and its anti-inflammatory and activity was dose-dependent and prolonged by niosomal encapsulation. It has also shown to promote collagen type 1 production in cell culture. Thus, AC could be a potential candidate for topical anti-inflammatory agent from natural resources.

Viscoelastic Modeling of Brain MRE Data Using FE Method

Dynamic shear test on simulated phantom can be used to validate magnetic resonance elastography (MRE) measurements. Phantom gel has been usually utilized for the cell culture of cartilage and soft tissue and also been used for mechanical property characterization using imaging systems. The viscoelastic property of the phantom would be important for dynamic experiments and analyses. In this study, An axisymmetric FE model is presented for determining the dynamic shear behaviour of brain simulated phantom using ABAQUS. The main objective of this study was to investigate the effect of excitation frequencies and boundary conditions on shear modulus and shear viscosity in viscoelastic media.

Plaque Formation of Toxoplasma gondii in Vero Cells using Carboxymethylcellulose

Toxoplasma gondii is an intracellular parasite capable of infecting all nucleated cells in a diverse array of species. Toxoplasma plaque assay have been described using Bacto Agar. Because of its experimental advantages carboxymethyl cellulose overlay, medium viscosity was choosing and the aim of this work was to develop alternative method for formation of T. gondii plaques. Tachyzoites were inoculated onto monolayers of Vero cells and cultured at 37° C under 5 % CO2. The cultures were followed up by microscopy inspection. Small plaques were visible by naphtol blue stain 4 days after infection. Larger plaques could be observed by day 10 of culture. The carboxymethyl cellulose is a cheap reagent and the methodology is easier, faster than assays under agar overlay. This is the first description of the carboxymethyl cellulose overlay use for obtaining the formation of T. gondii plaques and may be useful in consequent obtaining tachyzoites for detailed studies.

Multichannel Image Mosaicing of Stem Cells

Image mosaicing techniques are usually employed to offer researchers a wider field of view of microscopic image of biological samples. a mosaic is commonly achieved using automated microscopes and often with one “color" channel, whether it refers to natural or fluorescent analysis. In this work we present a method to achieve three subsequent mosaics of the same part of a stem cell culture analyzed in phase contrast and in fluorescence, with a common non-automated inverted microscope. The mosaics obtained are then merged together to mark, in the original contrast phase images, nuclei and cytoplasm of the cells referring to a mosaic of the culture, rather than to single images. The experiments carried out prove the effectiveness of our approach with cultures of cells stained with calcein (green/cytoplasm and nuclei) and hoechst (blue/nuclei) probes.

On-line Image Mosaicing of Live Stem Cells

Image mosaicing is a technique that permits to enlarge the field of view of a camera. For instance, it is employed to achieve panoramas with common cameras or even in scientific applications, to achieve the image of a whole culture in microscopical imaging. Usually, a mosaic of cell cultures is achieved through using automated microscopes. However, this is often performed in batch, through CPU intensive minimization algorithms. In addition, live stem cells are studied in phase contrast, showing a low contrast that cannot be improved further. We present a method to study the flat field from live stem cells images even in case of 100% confluence, this permitting to build accurate mosaics on-line using high performance algorithms.

Application of an in vitro Alveolus Model in Evaluating the Alveolar Response to Pressure- Induced Injury

In an effort to understand the preliminary effects of aerodynamic stress on alveolar epithelial cells, we developed a multifluidic cell culture platform capable of supporting alveolar cultures at an air-liquid interface under constant air flow and exposure to varying pressure stimuli on the apical side while providing nourishment on the basolateral plane. Our current study involved utilizing the platform to study the effect of basement membrane coating and addition of dexamethasone on cellular response to pressure in A549 and H441 alveolar epithelial cells.

Nonlinear Control of a Continuous Bioreactor Based on Cell Population Model

Saccharomyces cerevisiae (baker-s yeast) can exhibit sustained oscillations during the operation in a continuous bioreactor that adversely affects its stability and productivity. Because of heterogeneous nature of cell populations, the cell population balance models can be used to capture the dynamic behavior of such cultures. In this paper an unstructured, segregated model is used which is based on population balance equation(PBE) and then in order to simulation, the 4th order Rung-Kutta is used for time dimension and three methods, finite difference, orthogonal collocation on finite elements and Galerkin finite element are used for discretization of the cell mass domain. The results indicate that the orthogonal collocation on finite element not only is able to predict the oscillating behavior of the cell culture but also needs much little time for calculations. Therefore this method is preferred in comparison with other methods. In the next step two controllers, a globally linearizing control (GLC) and a conventional proportional-integral (PI) controller are designed for controlling the total cell mass per unit volume, and performances of these controllers are compared through simulation. The results show that although the PI controller has simpler structure, the GLC has better performance.