Search results for: in vitro propagation

Authors: Salma H. Abu Hafsa, A. Mendonca, B. Brehm-Stecher, A. A. Hassan, S. A. Ibrahim

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Enterococci are important inhabitants of the animal intestine and are widely used in probiotic products. A probiotic strain is expected to possess several desirable properties in order to exert beneficial effects. Therefore, the objective of this study was to isolate, characterize and identify Enterococcus sp. from chicken cecal and fecal samples to determine potential probiotic properties. Enterococci were isolated from chicken ceca and feces of thirty three clinically healthy chickens from a local farm. In vitro studies were performed to assess antibacterial activity of the isolated LAB (using agar well diffusion and cell free supernatant broth technique against Salmonella enterica serotype Enteritidis), survival in acidic conditions, resistance to bile salts, and their survival during simulated gastric juice conditions at pH 2.5. Isolates were identified by biochemical carbohydrate fermentation patterns using an API 50 CHL kit and API ZYM kits and by sequenced 16S rDNA. An isolate belonging to E. faecium species exhibited inhibitory effect against S. enteritidis. This isolate producing a clear zone as large as 10.30 mm or greater and was able to grow in the coculture medium and at the same time, inhibited the growth S. enteritidis. In addition, E. faecium exhibited significant resistance under highly acidic conditions at pH 2.5 for 8 h and survived well in bile salt at 0.2% for 24 h and showing ability to survive in the presence of simulated gastric juice at pH 2.5. Based on these results, E. faecium isolate fulfills some of the criteria to be considered as a probiotic strain and therefore, could be used as a feed additive with good potential for controlling S. Enteritidis in chickens. However, in vivo studies are needed to determine the safety of the strain.

Keywords: Acid tolerance, antimicrobial activity, Enterococcus faecium, probiotic.

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Authors: Addin Osman, Anwar Ali Yahya, Mohammed Basit Kamal

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Datasets or collections are becoming important assets by themselves and now they can be accepted as a primary intellectual output of a research. The quality and usage of the datasets depend mainly on the context under which they have been collected, processed, analyzed, validated, and interpreted. This paper aims to present a collection of program educational objectives mapped to student’s outcomes collected from self-study reports prepared by 32 engineering programs accredited by ABET. The manual mapping (classification) of this data is a notoriously tedious, time consuming process. In addition, it requires experts in the area, which are mostly not available. It has been shown the operational settings under which the collection has been produced. The collection has been cleansed, preprocessed, some features have been selected and preliminary exploratory data analysis has been performed so as to illustrate the properties and usefulness of the collection. At the end, the collection has been benchmarked using nine of the most widely used supervised multiclass classification techniques (Binary Relevance, Label Powerset, Classifier Chains, Pruned Sets, Random k-label sets, Ensemble of Classifier Chains, Ensemble of Pruned Sets, Multi-Label k-Nearest Neighbors and Back-Propagation Multi-Label Learning). The techniques have been compared to each other using five well-known measurements (Accuracy, Hamming Loss, Micro-F, Macro-F, and Macro-F). The Ensemble of Classifier Chains and Ensemble of Pruned Sets have achieved encouraging performance compared to other experimented multi-label classification methods. The Classifier Chains method has shown the worst performance. To recap, the benchmark has achieved promising results by utilizing preliminary exploratory data analysis performed on the collection, proposing new trends for research and providing a baseline for future studies.

Keywords: Benchmark collection, program educational objectives, student outcomes, ABET, Accreditation, machine learning, supervised multiclass classification, text mining.

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Authors: D. S. Zhuravkova, A. G. Temnikov, O. S. Belova, L. L. Chernensky, T. K. Gerastenok, I. Y. Kalugina, N. Y. Lysov, A.V. Orlov

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Investigation of the discharges from the artificial charged water aerosol clouds in presence of the arrays of the model hydrometeors could help to receive the new data about the peculiarities of the return stroke formation between the thundercloud and the ground when the large volumes of the hail particles participate in the lightning discharge initiation and propagation stimulation. Artificial charged water aerosol clouds of the negative or positive polarity with the potential up to one million volts have been used. Hail has been simulated by the group of the conductive model hydrometeors of the different form. Parameters of the impulse current of the main stage of the discharge between the artificial positively and negatively charged water aerosol clouds and the ground in presence of the model hydrometeors array and of its corresponding electromagnetic radiation have been determined. It was established that the parameters of the array of the model hydrometeors influence on the parameters of the main stage of the discharge between the artificial thundercloud cell and the ground. The maximal values of the main stage current impulse parameters and the electromagnetic radiation registered by the plate antennas have been found for the array of the model hydrometeors of the cylinder revolution form for the negatively charged aerosol cloud and for the array of the hydrometeors of the plate rhombus form for the positively charged aerosol cloud, correspondingly. It was found that parameters of the main stage of the discharge between the artificial charged water aerosol cloud and the ground in presence of the model hydrometeor array of the different considered forms depend on the polarity of the artificial charged aerosol cloud. In average, for all forms of the investigated model hydrometeors arrays, the values of the amplitude and the current rise of the main stage impulse current and the amplitude of the corresponding electromagnetic radiation for the artificial charged aerosol cloud of the positive polarity were in 1.1-1.9 times higher than for the charged aerosol cloud of the negative polarity. Thus, the received results could indicate to the possible more important role of the big volumes of the large hail arrays in the thundercloud on the parameters of the return stroke for the positive lightning.

Keywords: Main stage of discharge, hydrometeor form, lightning parameters, negative and positive artificial charged aerosol cloud.

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Authors: Fares A. Mayia, Mahmoud A. Yamany, Mushabbab A. Asiri

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In recent years, non-invasive Focused Ultrasound (FU) has been utilized for generating bubbles (cavities) to ablate target tissue by mechanical fractionation. Intensities >10 kW/cm² are required to generate the inertial cavities. The generation, rapid growth, and collapse of these inertial cavities cause tissue fractionation and the process is called Histotripsy. The ability to fractionate tissue from outside the body has many clinical applications including the destruction of the tumor mass. The process of tissue fractionation leaves a void at the treated site, where all the affected tissue is liquefied to particles at sub-micron size. The liquefied tissue will eventually be absorbed by the body. Histotripsy is a promising non-invasive treatment modality. This paper presents a technique for generating inertial cavities at lower intensities (< 1 kW/cm²). The technique (patent pending) is based on amplitude modulation (AM), whereby a low frequency signal modulates the amplitude of a higher frequency FU wave. Cavitation threshold is lower at low frequencies; the intensity required to generate cavitation in water at 10 kHz is two orders of magnitude lower than the intensity at 1 MHz. The Amplitude Modulation technique can operate in both continuous wave (CW) and pulse wave (PW) modes, and the percentage modulation (modulation index) can be varied from 0 % (thermal effect) to 100 % (cavitation effect), thus allowing a range of ablating effects from Hyperthermia to Histotripsy. Furthermore, changing the frequency of the modulating signal allows controlling the size of the generated cavities. Results from in vitro work demonstrate the efficacy of the new technique in fractionating soft tissue and solid calcium carbonate (Chalk) material. The technique, when combined with MR or Ultrasound imaging, will present a precise treatment modality for ablating diseased tissue without affecting the surrounding healthy tissue.

Keywords: Focused ultrasound therapy, Histotripsy, generation of inertial cavitation, mechanical tissue ablation.

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Authors: M. Saravanan

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Antimicrobial resistant is becoming a major factor in virtually all hospital acquired infection may soon untreatable is a serious public health problem. These concerns have led to major research effort to discover alternative strategies for the treatment of bacterial infection. Nanobiotehnology is an upcoming and fast developing field with potential application for human welfare. An important area of nanotechnology for development of reliable and environmental friendly process for synthesis of nanoscale particles through biological systems In the present studies are reported on the use of fungal strain Aspergillus species for the extracellular synthesis of bionanoparticles from 1 mM silver nitrate (AgNO3) solution. The report would be focused on the synthesis of metallic bionanoparticles of silver using a reduction of aqueous Ag+ ion with the culture supernatants of Microorganisms. The bio-reduction of the Ag+ ions in the solution would be monitored in the aqueous component and the spectrum of the solution would measure through UV-visible spectrophotometer The bionanoscale particles were further characterized by Atomic Force Microscopy (AFM), Fourier Transform Infrared Spectroscopy (FTIR) and Thin layer chromatography. The synthesized bionanoscale particle showed a maximum absorption at 385 nm in the visible region. Atomic Force Microscopy investigation of silver bionanoparticles identified that they ranged in the size of 250 nm - 680 nm; the work analyzed the antimicrobial efficacy of the silver bionanoparticles against various multi drug resistant clinical isolates. The present Study would be emphasizing on the applicability to synthesize the metallic nanostructures and to understand the biochemical and molecular mechanism of nanoparticles formation by the cell filtrate in order to achieve better control over size and polydispersity of the nanoparticles. This would help to develop nanomedicine against various multi drug resistant human pathogens.

Keywords: Bionanoparticles, UV-visible spectroscopy, AtomicForce Microscopy, Extracellular synthesis, Multi drug resistant, antimicrobial activity, Nanomedicine

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Authors: K. Kalaikumar, E. Baburaj

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One of the problems to be addressed in wireless sensor networks is the issues related to cross layer communication. Cross layer architecture shares the information across the layer, ensuring Quality of Services (QoS). With this shared information, MAC protocol adapts effective functionality maintenance such as route selection on changeable sensor network environment. However, time slot assignment and neighbour route selection time duration for cross layer have not been carried out. The time varying physical layer communication over cross layer causes high traffic load in the sensor network. Though, the traffic load was reduced using cross layer optimization procedure, the computational cost is high. To improve communication efficacy in the sensor network, a self-determined time slot based Cross-Layered Neighbour Route Discovery (C-LNRD) method is presented in this paper. In the presented work, the initial process is to discover the route in the sensor network using Dynamic Source Routing based Medium Access Control (MAC) sub layers. This process considers MAC layer operation with dynamic route neighbour table discovery. Then, the discovered route path for packet communication employs Broad Route Distributed Time Slot Assignment method on Cross-Layered Sensor Network system. Broad Route means time slotting on varying length of the route paths. During packet communication in this sensor network, transmission of packets is adjusted over the different time with varying ranges for controlling the traffic rate. Finally, Rayleigh fading model is developed in C-LNRD to identify the performance of the sensor network communication structure. The main task of Rayleigh Fading is to measure the power level of each communication under MAC sub layer. The minimized power level helps to easily reduce the computational cost of packet communication in the sensor network. Experiments are conducted on factors such as power factor, on packet communication, neighbour route discovery time, and information (i.e., packet) propagation speed.

Keywords: Medium access control, neighbour route discovery, wireless sensor network, Rayleigh fading, distributed time slot assignment

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Authors: Adel Dalilottojari, Bahman Delalat, Frances J. Harding, Michaelia P. Cockshell, Claudine S. Bonder, Nicolas H. Voelcker

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Endothelial Progenitor Cell (EPC) based therapies continue to be of interest to treat ischemic events based on their proven role to promote blood vessel formation and thus tissue re-vascularisation. Current strategies for the production of clinical-grade EPCs requires the in vitro isolation of EPCs from peripheral blood followed by cell expansion to provide sufficient quantities EPCs for cell therapy. This study aims to examine the use of different biomolecules to significantly improve the current strategy of EPC capture and expansion on collagen type I (Col I). In this study, four different biomolecules were immobilised on a surface and then investigated for their capacity to support EPC capture and proliferation. First, a cell microarray platform was fabricated by coating a glass surface with epoxy functional allyl glycidyl ether plasma polymer (AGEpp) to mediate biomolecule binding. The four candidate biomolecules tested were Col I, collagen type II (Col II), collagen type IV (Col IV) and vascular endothelial growth factor A (VEGF-A), which were arrayed on the epoxy-functionalised surface using a non-contact printer. The surrounding area between the printed biomolecules was passivated with polyethylene glycol-bisamine (A-PEG) to prevent non-specific cell attachment. EPCs were seeded onto the microarray platform and cell numbers quantified after 1 h (to determine capture) and 72 h (to determine proliferation). All of the extracellular matrix (ECM) biomolecules printed demonstrated an ability to capture EPCs within 1 h of cell seeding with Col II exhibiting the highest level of attachment when compared to the other biomolecules. Interestingly, Col IV exhibited the highest increase in EPC expansion after 72 h when compared to Col I, Col II and VEGF-A. These results provide information for significant improvement in the capture and expansion of human EPC for further application.

Keywords: Cardiovascular disease, cell microarray platform, cell therapy, endothelial progenitor cells, high throughput screening.

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Authors: Tomasz Dubiel, Tadeusz Balawender, Mirosław Osetek

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The paper presents the influence of non-metallic inclusions on the multi-stage forging process and the mechanical properties of the dodecagon socket bolt used in the automotive industry. The detected metallurgical defect was so large that it directly influenced the mechanical properties of the bolt and resulted in failure to meet the requirements of the mechanical property class. In order to assess the defect, an X-ray examination and metallographic examination of the defective bolt were performed, showing exogenous non-metallic inclusion. The size of the defect on the cross section was 0.531 mm in width and 1.523 mm in length; the defect was continuous along the entire axis of the bolt. In analysis, a finite element method (FEM) simulation of the multi-stage forging process was designed, taking into account a non-metallic inclusion parallel to the sample axis, reflecting the studied case. The process of defect propagation due to material upset in the head area was analyzed. The final forging stage in shaping the dodecagonal socket and filling the flange area was particularly studied. The effect of the defect was observed to significantly reduce the effective cross-section as a result of the expansion of the defect perpendicular to the axis of the bolt. The mechanical properties of products with and without the defect were analyzed. In the first step, the hardness test confirmed that the required value for the mechanical class 8.8 of both bolt types was obtained. In the second step, the bolts were subjected to a static tensile test. The bolts without the defect gave a positive result, while all 10 bolts with the defect gave a negative result, achieving a tensile strength below the requirements. Tensile strength tests were confirmed by metallographic tests and FEM simulation with perpendicular inclusion spread in the area of the head. The bolts were damaged directly under the bolt head, which is inconsistent with the requirements of ISO 898-1. It has been shown that non-metallic inclusions with orientation in accordance with the axis of the bolt can directly cause loss of functionality and these defects should be detected even before assembling in the machine element.

Keywords: continuous casting, multi-stage forging, non-metallic inclusion, upset bolt head

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Authors: Rawia M. Khalil, Ahmed A. Abd El Rahman, Mahfouz A. Kassem, Mohamed S. El Ridi, Mona M. Abou Samra, Ghada E. A. Awad, Soheir S. Mansy

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Solid lipid nanoparticles (SLNs) have gained great attention for the topical treatment of skin associated fungal infection as they facilitate the skin penetration of loaded drugs. Our work deals with the preparation of nystatin loaded solid lipid nanoparticles (NystSLNs) using the hot homogenization and ultrasonication method. The prepared NystSLNs were characterized in terms of entrapment efficiency, particle size, zeta potential, transmission electron microscopy, differential scanning calorimetry, rheological behavior and in vitro drug release. A stability study for 6 months was performed. A microbiological study was conducted in male rats infected with Candida albicans, by counting the colonies and examining the histopathological changes induced on the skin of infected rats. The results showed that SLNs dispersions are spherical in shape with particle size ranging from 83.26±11.33 to 955.04±1.09 nm. The entrapment efficiencies are ranging from 19.73±1.21 to 72.46±0.66% with zeta potential ranging from -18.9 to -38.8 mV and shear-thinning rheological Behavior. The stability studies done for 6 months showed that nystatin (Nyst) is a good candidate for topical SLN formulations. A least number of colony forming unit/ ml (cfu/ml) was recorded for the selected NystSLN compared to the drug solution and the commercial Nystatin® cream present in the market. It can be fulfilled from this work that SLNs provide a good skin targeting effect and may represent promising carrier for topical delivery of Nyst offering the sustained release and maintaining the localized effect, resulting in an effective treatment of cutaneous fungal infection.

Keywords: Candida infections, Hot homogenization, Nystatin, Solid lipid nanoparticles, Stability, Topical delivery.

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Authors: H. M. Nanjundaswamy, S. K. Nath, S. Ray

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TiO₂ particles have been added in molten aluminium to result in aluminium based cast Al/Al₃Ti-Al₂O₃ composite, which has been added then to molten magnesium to synthesize magnesium based cast Mg-Al/Al₃Ti-Al₂O₃ composite. The nominal compositions in terms of Mg, Al, and TiO₂ contents in the magnesium based composites are Mg-9Al-0.6TiO₂, Mg-9Al-0.8TiO₂, Mg-9Al-1.0TiO₂ and Mg-9Al-1.2TiO₂ designated respectively as MA6T, MA8T, MA10T and MA12T. The microstructure of the cast magnesium based composite shows grayish rods of intermetallics Al₃Ti, inherited from aluminium based composite but these rods, on hot forging, breaks into smaller lengths decreasing the average aspect ratio (length to diameter) from 7.5 to 3.0. There are also cavities in between the broken segments of rods. β-phase in cast microstructure, Mg₁₇Al₁₂, dissolves during heating prior to forging and re-precipitates as relatively finer particles on cooling. The amount of β-phase also decreases on forging as segregation is removed. In both the cast and forged composite, the Brinell hardness increases rapidly with increasing addition of TiO₂ but the hardness is higher in forged composites by about 80 BHN. With addition of higher level of TiO₂ in magnesium based cast composite, yield strength decreases progressively but there is marginal increase in yield strength over that of the cast Mg-9 wt. pct. Al, designated as MA alloy. But the ultimate tensile strength (UTS) in the cast composites decreases with the increasing particle content indicating possibly an early initiation of crack in the brittle inter-dendritic region and their easy propagation through the interfaces of the particles. In forged composites, there is a significant improvement in both yield strength and UTS with increasing TiO₂ addition and also, over those observed in their cast counterpart, but at higher addition it decreases. It may also be noted that as in forged MA alloy, incomplete recovery of forging strain increases the strength of the matrix in the composites and the ductility decreases both in the forged alloy and the composites. Initiation fracture toughness, J_IC, decreases drastically in cast composites compared to that in MA alloy due to the presence of intermetallic Al₃Ti and Al₂O₃ particles in the composite. There is drastic reduction of J_IC on forging both in the alloy and the composites, possibly due to incomplete recovery of forging strain in both as well as breaking of Al₃Ti rods and the voids between the broken segments of Al₃Ti rods in composites. The ratio of tearing modulus to elastic modulus in cast composites show higher ratio, which increases with the increasing TiO₂ addition. The ratio decreases comparatively more on forging of cast MA alloy than those in forged composites.

Keywords: Composite, fracture toughness, forging, tensile properties.

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Authors: A. M. Alaskari, S. E. Oraby

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The quality of a machined surface is becoming more and more important to justify the increasing demands of sophisticated component performance, longevity, and reliability. Usually, any machining operation leaves its own characteristic evidence on the machined surface in the form of finely spaced micro irregularities (surface roughness) left by the associated indeterministic characteristics of the different elements of the system: tool-machineworkpart- cutting parameters. However, one of the most influential sources in machining affecting surface roughness is the instantaneous state of tool edge. The main objective of the current work is to relate the in-process immeasurable cutting edge deformation and surface roughness to a more reliable easy-to-measure force signals using a robust non-linear time-dependent modeling regression techniques. Time-dependent modeling is beneficial when modern machining systems, such as adaptive control techniques are considered, where the state of the machined surface and the health of the cutting edge are monitored, assessed and controlled online using realtime information provided by the variability encountered in the measured force signals. Correlation between wear propagation and roughness variation is developed throughout the different edge lifetimes. The surface roughness is further evaluated in the light of the variation in both the static and the dynamic force signals. Consistent correlation is found between surface roughness variation and tool wear progress within its initial and constant regions. At the first few seconds of cutting, expected and well known trend of the effect of the cutting parameters is observed. Surface roughness is positively influenced by the level of the feed rate and negatively by the cutting speed. As cutting continues, roughness is affected, to different extents, by the rather localized wear modes either on the tool nose or on its flank areas. Moreover, it seems that roughness varies as wear attitude transfers from one mode to another and, in general, it is shown that it is improved as wear increases but with possible corresponding workpart dimensional inaccuracy. The dynamic force signals are found reasonably sensitive to simulate either the progressive or the random modes of tool edge deformation. While the frictional force components, feeding and radial, are found informative regarding progressive wear modes, the vertical (power) components is found more representative carrier to system instability resulting from the edge-s random deformation.

Keywords: Dynamic force signals, surface roughness (finish), tool wear and deformation, tool wear modes (nose, flank)

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Authors: Siddhartha Bhattacharyya, Paramartha Dutta, Ujjwal Maulik, Prashanta Kumar Nandi

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A multilayer self organizing neural neural network (MLSONN) architecture for binary object extraction, guided by a beta activation function and characterized by backpropagation of errors estimated from the linear indices of fuzziness of the network output states, is discussed. Since the MLSONN architecture is designed to operate in a single point fixed/uniform thresholding scenario, it does not take into cognizance the heterogeneity of image information in the extraction process. The performance of the MLSONN architecture with representative values of the threshold parameters of the beta activation function employed is also studied. A three layer bidirectional self organizing neural network (BDSONN) architecture comprising fully connected neurons, for the extraction of objects from a noisy background and capable of incorporating the underlying image context heterogeneity through variable and adaptive thresholding, is proposed in this article. The input layer of the network architecture represents the fuzzy membership information of the image scene to be extracted. The second layer (the intermediate layer) and the final layer (the output layer) of the network architecture deal with the self supervised object extraction task by bi-directional propagation of the network states. Each layer except the output layer is connected to the next layer following a neighborhood based topology. The output layer neurons are in turn, connected to the intermediate layer following similar topology, thus forming a counter-propagating architecture with the intermediate layer. The novelty of the proposed architecture is that the assignment/updating of the inter-layer connection weights are done using the relative fuzzy membership values at the constituent neurons in the different network layers. Another interesting feature of the network lies in the fact that the processing capabilities of the intermediate and the output layer neurons are guided by a beta activation function, which uses image context sensitive adaptive thresholding arising out of the fuzzy cardinality estimates of the different network neighborhood fuzzy subsets, rather than resorting to fixed and single point thresholding. An application of the proposed architecture for object extraction is demonstrated using a synthetic and a real life image. The extraction efficiency of the proposed network architecture is evaluated by a proposed system transfer index characteristic of the network.

Keywords: Beta activation function, fuzzy cardinality, multilayer self organizing neural network, object extraction,

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Authors: Rajan Rajabalaya, Li-Qun Tor, Sheba David

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Transdermal delivery of ondansetron hydrochloride (OdHCl) can prevent the problems encountered with oral ondansetron. In previously conducted studies, effect of amount of polyvinyl pyrrolidone, permeation enhancer and casting solvent on the physicochemical properties on OdHCl were investigated. It is feasible to develop ondansetron transdermal patch by using ethyl cellulose and polyvinyl pyrrolidone with dibutyl pthalate as plasticizer, however, the desired flux is not achieved. The primary aim of this study is to use dimethyl succinate (DMS) and propylene glycol that are not incorporated in previous studies to determine their effect on the physicochemical properties of an OdHCl transdermal patch using ethyl cellulose and polyvinyl pyrrolidone. This study also investigates the effect of permeation enhancer (eugenol and phosphatidylcholine) on the release of OdHCl. The results showed that propylene glycol is a more suitable plasticizer compared to DMS in the fabrication of OdHCl transdermal patch using ethyl cellulose and polyvinyl pyrrolidone as polymers. Propylene glycol containing patch has optimum drug content, thickness, moisture content and water absorption, tensile strength, and a better release profile than DMS. Eugenol and phosphatidylcholine can increase release of OdHCl from the patches. From the physicochemical result and permeation profile, a combination of 350mg of ethyl cellulose, 150mg polyvinyl pyrrolidone, 3% of total polymer weight of eugenol, and 40% of total polymer weight of propylene glycol is the most suitable formulation to develop an OdHCl patch. OdHCl release did not increase with increasing the percentage of plasticiser. DMS 4, PG 4, DMS 9, PG 9, DMS 14, and PG 14 gave better release profiles where using 300mg: 0mg, 300mg: 100mg, and 350mg: 150mg of EC: PVP. Thus, 40% of PG or DMS appeared to be the optimum amount of plasticiser when the above combination where EC: PVP was used. It was concluded from the study that a patch formulation containing 350mg EC, 150mg PVP, 40% PG and 3% eugenol is the best transdermal matrix patch compositions for the uniform and continuous release/permeation of OdHCl over an extended period. This patch design can be used for further pharmacokinetic and pharmacodynamic studies in suitable animal models.

Keywords: Ondansetron hydrochloride, dimethyl succinate, eugenol.

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Authors: Jienny Lee, In-Soo Cho, Sang-Ho Cha

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Adult mesenchymal stem cells (MSCs) have been investigated using preclinical approaches for tissue regeneration. Porcine MSCs (pMSCs) are capable of growing and attaching to plastic with a fibroblast-like morphology and then differentiating into bone, adipose, and cartilage tissues in vitro. This study was conducted to investigate the proliferating abilities, differentiation potentials, and multipotency of miniature pig adipose tissue-derived MSCs (mpAD-MSCs) with or without long-term cryopreservation, considering that cryostorage has the potential for use in clinical applications. After confirming the characteristics of the mpAD-MSCs, we examined the effect of long-term cryopreservation (> 2 years) on expression of cell surface markers (CD34, CD90 and CD105), proliferating abilities (cumulative population doubling level, doubling time, colony-forming unit, and MTT assay) and differentiation potentials into mesodermal cell lineages. As a result, the expression of cell surface markers is similar between thawed and fresh mpAD-MSCs. However, long-term cryopreservation significantly lowered the differentiation potentials (adipogenic, chondrogenic, and osteogenic) of mpAD-MSCs. When compared with fresh mpAD-MSCs, thawed mpAD-MSCs exhibited lower expression of mesodermal cell lineage-related genes such as peroxisome proliferator-activated receptor-g2, lipoprotein lipase, collagen Type II alpha 1, osteonectin, and osteocalcin. Interestingly, long-term cryostoraged mpAD-MSCs exhibited significantly higher cell viability than the fresh mpAD-MSCs. Long-term cryopreservation induced a 30% increase in the cell viability of mpAD-MSCs when compared with the fresh mpAD-MSCs at 5 days after thawing. However, long-term cryopreservation significantly lowered expression of stemness markers such as Oct3/4, Sox2, and Nanog. Furthermore, long-term cryopreservation negatively affected expression of senescence-associated genes such as telomerase reverse transcriptase and heat shock protein 90 of mpAD-MSCs when compared with the fresh mpAD-MSCs. The results from this study might be important for the successful application of MSCs in clinical trials after long-term cryopreservation.

Keywords: Mesenchymal stem cells, Cryopreservation, Stemness, Senescence.

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Authors: Ibrahim M. S. Shnawa, Bashar A. H. E. Alsadi, Kalida K. Alniaem

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In the immunologic sense, clinical infection is a state of failure of the immune system to combat the pathogenic weapon of the bacteria invading the host. A motile gram negative vibroid organism associated with marked mono and poly nuclear cell responses was traced during the examination of a clinical material from an infected common carp Cyprinus carpio. On primary plate culture, growth was shown to be pure, dense population of an Aeromonas-like colony morphotype. The pure isolate was found to be; Aerobic, facultatively anaerobic, non-halophilic, grew at 0C, and 37C, oxidase positive utilizes glucose through fermentative pathway, resist 0/129 and novobiocin, produces alanine and lysine decarboxylases but non-producing ornithine dehydrolases. Tests for the in vitro determinants of pathogenicity has shown to be; Betahaemolytic onto blood agar, gelatinase, casienase and amylase producer. Three in vivo determinants of pathogenicity were tested as, the lethal dose fifty, the pathogenesis and pathogenicity. It was evident that 0.1 milliliter of the causal bacterial cell suspension of a density 1 x 107 CFU/ml injected intramuscularly into an average of 100gms fish toke five days incubation period, then at the day six morbidity and mortality were initiated. LD50 was recorded at the day 12 post-infection. Use of an LD50 doses to study the pathogenicity, reveals mononuclear and polynuclear cell responses, on examining the stained direct films of the clinical materials from the experimentally infected fish. Re-isolation tests confirm that the reisolant is same. The course of the infection in natural case was shown manifestation of; skin ulceration, haemorrhage and descaling. On evisceration, the internal organs were shown; congestion in the intestines, spleen and, air sacs. The induced infection showed a milder form of these manifestations. The grading of the virulence of this organism was virulent causing chronic course of infections as indicated from the pathogenesis and pathogenicity studies. Thus the infectious bacteria were consistent with Aeromonas hydrophila, and the infection was chronic.

Keywords: Piscan, inflammatory respnonse, pure culture, pathogen, chronic, infection.

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Authors: A. Dashti, M. Eskandari, L. Farahmand, P. Parvin, A. Jafargholi

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Breast Cancer is one of the most considerable diseases in the United States and other countries and is the second leading cause of death in women. Common breast cancer treatments would lead to adverse side effects such as loss of hair, nausea, and weakness. These complications arise because these cancer treatments damage some healthy cells while eliminating the cancer cells. In an effort to address these complications, laser radiation was utilized and tested as a targeted cancer treatment for breast cancer. In this regard, tissue engineering approaches are being employed by using an electrospun scaffold in order to facilitate the growth of breast cancer cells. Polycaprolacton (PCL) was used as a material for scaffold fabricating because of its biocompatibility, biodegradability, and supporting cell growth. The specific breast cancer cells have the ability to create a three-dimensional cell cluster due to the spontaneous accumulation of cells in the porosity of the scaffold under some specific conditions. Therefore, we are looking for a higher density of porosity and larger pore size. Fibers showed uniform diameter distribution and final scaffold had optimum characteristics with approximately 40% porosity. The images were taken by SEM and the density and the size of the porosity were determined with the Image. After scaffold preparation, it has cross-linked by glutaraldehyde. Then, it has been washed with glycine and phosphate buffer saline (PBS), in order to neutralize the residual glutaraldehyde. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromidefor (MTT) results have represented approximately 91.13% viability of the scaffolds for cancer cells. In order to create a cluster, Michigan Cancer Foundation-7 (MCF-7, breast cancer cell line) and Michigan Cancer Foundation-10A (MCF-10A, human mammary epithelial cell line) cells were cultured on the scaffold in 24 well plate for five days. Then, we have exposed the cluster to the laser diode 808 nm radiation to investigate the effect of laser on the tumor with different power and time. Under the same conditions, cancer cells lost their viability more than the healthy ones. In conclusion, laser therapy is a viable method to destroy the target cells and has a minimum effect on the healthy tissues and cells and it can improve the other method of cancer treatments limitations.

Keywords: Breast cancer, electrospun scaffold, polycaprolacton, laser diode, cancer treatment.

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Authors: Amlan Kumar Das, Avinash Marwal, Vikram Pareek

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Liposome plays an important role in medical and pharmaceutical science as e.g. nano scale drug carriers. Liposomes are vesicles of varying size consisting of a spherical lipid bilayer and an aqueous inner compartment. Magnet-driven liposome used for the targeted delivery of drugs to organs and tissues. These liposome preparations contain encapsulated drug components and finely dispersed magnetic particles. Liposomes are vesicles of varying size consisting of a spherical lipid bilayer and an aqueous inner compartment that are generated in vitro. These are useful in terms of biocompatibility, biodegradability, and low toxicity, and can control biodistribution by changing the size, lipid composition, and physical characteristics. Furthermore, liposomes can entrap both hydrophobic and hydrophilic drugs and are able to continuously release the entrapped substrate, thus being useful drug carriers. Magnetic liposomes (MLs) are phospholipid vesicles that encapsulate magneticor paramagnetic nanoparticles. They are applied as contrast agents for magnetic resonance imaging (MRI). The biological synthesis of nanoparticles using plant extracts plays an important role in the field of nanotechnology. Green-synthesized magnetite nanoparticles-protein hybrid has been produced by treating Iron (III) / Iron (II) chloride with the leaf extract of Datura inoxia. The phytochemicals present in the leaf extracts act as a reducing as well stabilizing agents preventing agglomeration, which include flavonoids, phenolic compounds, cardiac glycosides, proteins and sugars. The magnetite nanoparticles-protein hybrid has been trapped inside the aqueous core of the liposome prepared by reversed phase evaporation (REV) method using oleic and linoleic acid which has been shown to be driven under magnetic field confirming the formation magnetic liposome (ML). Chemical characterization of stealth magnetic liposome has been performed by breaking the liposome and release of magnetic nanoparticles. The presence iron has been confirmed by colour complex formation with KSCN and UV-Vis study using spectrophotometer Cary 60, Agilent. This magnet driven liposome using nanoparticles-protein hybrid can be a smart vesicles for the targeted drug delivery.

Keywords: Nanoparticles-Protein Hybrid, Magnetic Liposome.

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Authors: Aliya Isaeva, Anar Sultaniarova

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The report focuses on such an important indicator of the nature and direction of development of ethnic and cultural processes in the Republic of Kazakhstan, as ethno linguistic situation. It is shown that, in essence, on the one hand, expresses the degree of the actual propagation and the level of use of the languages of the various ethnic communities. On the other hand, reflects the important patterns, trends and prospects of ethno-cultural and ethnodemographic processes in the Republic. It is important to note that the ethno linguistic situation in different regions of Kazakhstan, due to its more dynamic and much more difficult to demonstrate a much greater variety of options when compared with the ethnic situation in the country. For the two major ethnic groups of the republic – Kazakh and Russian language ethno differentiating retains its value, while for the other ethnic groups observed decline in the importance of this indicator. As you know, the language of international communication in the country is Russian. As the censuses of population, the Russian language in many areas of Northern, Central and Eastern Kazakhstan becomes a means of ethno linguistic development for most of the non-Russian population. This is most clearly illustrated by the Germans, and the Slavic ethnic groups. In this case, the Russian language is not just a means of international communication for a number of ethnic groups, and ethnic groups, it becomes a factor of ethnic self-expression. The value of the Kazakh language as their mother tongue for the other groups of the population is small. More clearly it can be traced only to the Turkic-speaking population of the republic – Uzbeks, Uighurs, Tatars, Turks, etc. The state Kazakh language is a means of international communication in the Western and Southern Kazakhstan, with a predominance of the Kazakh population. The report shows that the most important factor in the development of ethno-linguistic and ethno-cultural processes is bilingualism. Comparative analysis of materials census shows, first, on the increase of the proportion of bilingual population among Kazakhs and Russian, and second, to reduce the proportion of bilingual population of other ethnic groups living in Kazakhstan, and third, a higher proportion bilingual population among residents than rural residents, regardless of their ethnicity. Bilingualism is mainly of a "national Kazakh", "national Russian" or "Kazakh-national" or "Russian-national" character. The President N.A. Nazarbayev said that the Kazakh language is the most important factor in the consolidation of the people of Kazakhstan. He therefore called on government and other state and local representative bodies fully develop the state language, to create all the necessary organizational, material and technical conditions for free and open learning the state language by all citizens of the Republic of Kazakhstan.

Keywords: Ethnos, ethno cultural processes, ethnolinguistic situation, mother tongue, bilingualism.

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Authors: Naira Sahakyan, Margarit Petrosyan, Armen Trchounian

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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.

Keywords: Ajuga genevensis, antibacterial activity, antioxidant activity, callus cultures.

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Authors: S. Mehrab Amiri, Nasser Talebbeydokhti

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Modeling sediment transport processes by means of numerical approach often poses severe challenges. In this way, a number of techniques have been suggested to solve flow and sediment equations in decoupled, semi-coupled or fully coupled forms. Furthermore, in order to capture flow discontinuities, a number of techniques, like artificial viscosity and shock fitting, have been proposed for solving these equations which are mostly required careful calibration processes. In this research, a numerical scheme for solving shallow water and Exner equations in fully coupled form is presented. First-Order Centered scheme is applied for producing required numerical fluxes and the reconstruction process is carried out toward using Monotonic Upstream Scheme for Conservation Laws to achieve a high order scheme.  In order to satisfy C-property of the scheme in presence of bed topography, Surface Gradient Method is proposed. Combining the presented scheme with fourth order Runge-Kutta algorithm for time integration yields a competent numerical scheme. In addition, to handle non-prismatic channels problems, Cartesian Cut Cell Method is employed. A trained Multi-Layer Perceptron Artificial Neural Network which is of Feed Forward Back Propagation (FFBP) type estimates sediment flow discharge in the model rather than usual empirical formulas. Hydrodynamic part of the model is tested for showing its capability in simulation of flow discontinuities, transcritical flows, wetting/drying conditions and non-prismatic channel flows. In this end, dam-break flow onto a locally non-prismatic converging-diverging channel with initially dry bed conditions is modeled. The morphodynamic part of the model is verified simulating dam break on a dry movable bed and bed level variations in an alluvial junction. The results show that the model is capable in capturing the flow discontinuities, solving wetting/drying problems even in non-prismatic channels and presenting proper results for movable bed situations. It can also be deducted that applying Artificial Neural Network, instead of common empirical formulas for estimating sediment flow discharge, leads to more accurate results.

Keywords: Artificial neural network, morphodynamic model, sediment continuity equation, shallow water equations.

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Authors: T. Garlan, I. Gabelotaud, S. Lucas, E. Marchès

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Production of a global sedimentological seabed map has been initiated in 1995 to provide the necessary tool for searches of aircraft and boats lost at sea, to give sedimentary information for nautical charts, and to provide input data for acoustic propagation modelling. This original approach had already been initiated one century ago when the French hydrographic service and the University of Nancy had produced maps of the distribution of marine sediments of the French coasts and then sediment maps of the continental shelves of Europe and North America. The current map of the sediment of oceans presented was initiated with a UNESCO's general map of the deep ocean floor. This map was adapted using a unique sediment classification to present all types of sediments: from beaches to the deep seabed and from glacial deposits to tropical sediments. In order to allow good visualization and to be adapted to the different applications, only the granularity of sediments is represented. The published seabed maps are studied, if they present an interest, the nature of the seabed is extracted from them, the sediment classification is transcribed and the resulted map is integrated in the world map. Data come also from interpretations of Multibeam Echo Sounder (MES) imagery of large hydrographic surveys of deep-ocean. These allow a very high-quality mapping of areas that until then were represented as homogeneous. The third and principal source of data comes from the integration of regional maps produced specifically for this project. These regional maps are carried out using all the bathymetric and sedimentary data of a region. This step makes it possible to produce a regional synthesis map, with the realization of generalizations in the case of over-precise data. 86 regional maps of the Atlantic Ocean, the Mediterranean Sea, and the Indian Ocean have been produced and integrated into the world sedimentary map. This work is permanent and permits a digital version every two years, with the integration of some new maps. This article describes the choices made in terms of sediment classification, the scale of source data and the zonation of the variability of the quality. This map is the final step in a system comprising the Shom Sedimentary Database, enriched by more than one million punctual and surface items of data, and four series of coastal seabed maps at 1:10,000, 1:50,000, 1:200,000 and 1:1,000,000. This step by step approach makes it possible to take into account the progresses in knowledge made in the field of seabed characterization during the last decades. Thus, the arrival of new classification systems for seafloor has improved the recent seabed maps, and the compilation of these new maps with those previously published allows a gradual enrichment of the world sedimentary map. But there is still a lot of work to enhance some regions, which are still based on data acquired more than half a century ago.

Keywords: Marine sedimentology, seabed map, sediment classification, World Ocean.

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Authors: Benjawan Saengwichian, Alasdair E. Charles, Philip J. Hyde

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Magnetically controlled growing rods (MCGRs) have been used to stabilise and correct spinal curvature in children to support non-invasive scoliosis adjustment. Although the encapsulated driving components are intended to be isolated from body fluid contact, in vivo corrosion was observed on these components due to sealing mechanism damage. Consequently, a corrosion circuit is created with the body fluids, resulting in malfunction of the lengthening mechanism. Particularly, the chloride ions in blood plasma or cerebrospinal fluid (CSF) may corrode the MCGR alloys, possibly resulting in metal ion release in long-term use. However, there is no data available on the corrosion resistance of spinal implant alloys in CSF. In this study, an in vitro immersion configuration was designed to simulate in vivo corrosion of 440C SS-Ti6Al4V couples. The 440C stainless steel (SS) was heat-treated to investigate the effect of tempering temperature on intergranular corrosion (IGC), while crevice and galvanic corrosion were studied by limiting the clearance of dissimilar couples. Tests were carried out in a neutral artificial cerebrospinal fluid (ACSF) and phosphate-buffered saline (PBS) under aeration and deaeration for 2 months. The composition of the passive films and metal ion release were analysed. The effect of galvanic coupling, pH, dissolved oxygen and anion species on corrosion rates and corrosion mechanisms are discussed based on quantitative and qualitative measurements. The results suggest that ACSF is more aggressive than PBS due to the combination of aggressive chlorides and sulphate anions, while phosphate in PBS acts as an inhibitor to delay corrosion. The presence of Vivianite on the SS surface in PBS lowered the corrosion rate (CR) more than 5 times for aeration and nearly 2 times for deaeration, compared with ACSF. The CR of 440C is dependent on passive film properties varied by tempering temperature and anion species. Although the CR of Ti6Al4V is insignificant, it tends to release more Ti ions in deaerated ACSF than under aeration, about 6 µg/L. It seems the crevice-like design has more effect on macroscopic corrosion than combining the dissimilar couple, whereas IGC is dominantly observed on sensitized microstructure.

Keywords: Cerebrospinal fluid, crevice corrosion, intergranular corrosion, magnetically controlled growing rods.

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Authors: Ibtisam A. Abbas Al-Darkazly

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In this study, the mechanical properties of alginate hydrogel material for self-standing 3D scaffold architecture with proper shape fidelity are investigated. In-lab built 3D bio-printer extrusion-based technology is utilized to fabricate 3D alginate scaffold constructs. The pressure, needle speed and stage speed are varied using a computer-controlled system. The experimental result indicates that the concentration of alginate solution, calcium chloride (CaCl₂) cross-linking concentration and cross-linking ratios lead to the formation of alginate hydrogel with various gelation states. Besides, the gelling conditions, such as cross-linking reaction time and temperature also have a significant effect on the mechanical properties of alginate hydrogel. Various experimental tests such as the material gelation, the material spreading and the printability test for filament collapse as well as the swelling test were conducted to evaluate the fabricated 3D scaffold constructs. The result indicates that the fabricated 3D scaffold from composition of 3.5% wt alginate solution, that is prepared in DI water and 1% wt CaCl₂ solution with cross-linking ratios of 7:3 show good printability and sustain good shape fidelity for more than 20 days, compared to alginate hydrogel that is prepared in a phosphate buffered saline (PBS). The fabricated self-standing 3D scaffold constructs measured 30 mm × 30 mm and consisted of 4 layers (n = 4) show good pore geometry and clear grid structure after printing. In addition, the percentage change of swelling degree exhibits high swelling capability with respect to time. The swelling test shows that the geometry of 3D alginate-scaffold construct and of the macro-pore are rarely changed, which indicates the capability of holding the shape fidelity during the incubation period. This study demonstrated that the mechanical and physical properties of alginate hydrogel could be tuned for a 3D bio-printing extrusion-based system to fabricate self-standing 3D scaffold soft structures. This 3D bioengineered scaffold provides a natural microenvironment present in the extracellular matrix of the tissue, which could be seeded with the biological cells to generate the desired 3D live tissue model for in vitro and in vivo tissue engineering applications.

Keywords: Biomaterial, calcium chloride, 3D bio-printing, extrusion, scaffold, sodium alginate, tissue engineering.

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Authors: Samar Saadeldin Abdelmotalab Omer, Ikram Mohamed Eltayeb Elsiddig, Amna Beshir Medani Ahmed, Saad Mohamed Hussein Ayoub

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Many herbal medicinal products are considered potential anti-hypercholesterolemic agents with encouraging safety profiles, however only a limited amount of clinical research exists to support their efficacy. The present study was designed to compare the antihypercholesterolemic and antioxidant activities of the crude ethanolic extracts of Citrus reticulata fruit peel, Zingiber officinale rhizome and Sesamum indicum seeds. Forty-five rats were used throughout the experiment which are extended for four weeks. These were divided into nine groups, five rats per each group as follows; group 1 was the normal control group (rats only fed standard normal rat diet), group 2 was the hypercholesterolemic control group (rats fed only hypercholesterolemic diet which contained 1% cholesterol plus 10% saturated animal fat added to the normal rat diet), groups 3 and 4 were fed hypercholesterolemic diet in addition to Citrus reticulata ethanolic extract at doses of (250mg/kg (group 3) and 500mg/kg (group 4)) administered daily via oral route, groups 5 and 6 were given hypercholesterolemic diet in addition to Zingiber officinale ethanolic extract at doses of (250mg/kg (group 5) and 500mg/kg (group 6)) daily through oral route, groups 7 and 8 fed on hypercholesterolemic diet in addition to Sesamum indicum ethanolic extract at doses of (250mg/kg (group 7) and 500mg/kg (group 8)) daily orally; and group 9 rats were given hypercholesterolemic diet in addition to atorvastatin (0.18mg/kg) daily via oral route as a standard reference antihypercholesterolemic drug. Blood samples from all groups were drawn from the retro-orbital venous plexus four weeks following treatment after overnight fasting and the lipid profile (total cholesterol (TC), high density lipoprotein-cholesterol (HDL-C), low density lipoprotein-cholesterol (LDL-C) and triglyceride levels) were measured and the risk ratio (TC/HDL-C) was assessed. The antioxidant activity of the three plants extracts was determined using DPPH free-radical antioxidant assay. Results of in vivo and in vitro antihypercholesterolemic and antioxidant assay respectively, revealed that the three extracts possess comparable antioxidant and antihypercholesterolemic activities.

Keywords: Antihypercholesterolemic effects, Antioxidant activity, HDL, LDL, TC, TGs, Citrus reticulata, Sesamum indicum, Zingiber officinale.

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Authors: A. M. Tarola, R. Preti, A. M. Girelli, P. Campana

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Safeguarding, studying and enhancing biodiversity play an important and indispensable role in re-launching agriculture. The ancient local varieties are therefore a precious resource for genetic and health improvement. In order to protect biodiversity through the recovery and valorization of autochthonous varieties, in this study we analyzed 12 samples of four ancient apple cultivars representative of Friuli Venezia Giulia, selected by local farmers who work on a project for the recovery of ancient apple cultivars. The aim of this study is to evaluate the polyphenolic profile and the antioxidant capacity that characterize the organoleptic and functional qualities of this fruit species, besides having beneficial properties for health. In particular, for each variety, the following compounds were analyzed, both in the skins and in the pulp: gallic acid, catechin, chlorogenic acid, epicatechin, caffeic acid, coumaric acid, ferulic acid, rutin, phlorizin, phloretin and quercetin to highlight any differences in the edible parts of the apple. The analysis of individual phenolic compounds was performed by High Performance Liquid Chromatography (HPLC) coupled with a diode array UV detector (DAD), the antioxidant capacity was estimated using an in vitro essay based on a Free Radical Scavenging Method and the total phenolic compounds was determined using the Folin-Ciocalteau method. From the results, it is evident that the catechins are the most present polyphenols, reaching a value of 140-200 μg/g in the pulp and of 400-500 μg/g in the skin, with the prevalence of epicatechin. Catechins and phlorizin, a dihydrohalcone typical of apples, are always contained in larger quantities in the peel. Total phenolic compounds content was positively correlated with antioxidant activity in apple pulp (r² = 0,850) and peel (r² = 0,820). Comparing the results, differences between the varieties analyzed and between the edible parts (pulp and peel) of the apple were highlighted. In particular, apple peel is richer in polyphenolic compounds than pulp and flavonols are exclusively present in the peel. In conclusion, polyphenols, being antioxidant substances, have confirmed the benefits of fruit in the diet, especially as a prevention and treatment for degenerative diseases. They demonstrated to be also a good marker for the characterization of different apple cultivars. The importance of protecting biodiversity in agriculture was also highlighted through the exploitation of native products and ancient varieties of apples now forgotten.

Keywords: Apple, biodiversity, polyphenols, antioxidant activity, HPLC-DAD, characterization.

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Authors: M. Naimi, M. B. Khaled

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The aim of the present work was to test in vitro inhibition of food pathogens and spoilage bacteria by crude bacteriocins from autochthonous lactic acid bacteria. Thirty autochthonous lactic acid bacteria isolated previously, belonging to the genera: Lactobacillus, Carnobacterium, Lactococcus, Vagococcus, Streptococcus, and Pediococcus, have been screened by an agar spot test and a well diffusion assay against Gram-positive and Gram-negative harmful bacteria: Bacillus cereus, Bacillus subtilis ATCC 6633, Escherichia coli ATCC 8739, Salmonella typhimurium ATCC 14028, Staphylococcus aureus ATCC 6538, and Pseudomonas aeruginosa under conditions means to reduce lactic acid and hydrogen peroxide effect to select bacteria with high bacteriocinogenic potential. Furthermore, crude bacteriocins semiquantification and heat sensitivity to different temperatures (80, 95, 110°C, and 121°C) were performed. Another exploratory test concerning the response of St. aureus ATCC 6538 to the presence of crude bacteriocins was realized. It has been observed by the agar spot test that fifteen candidates were active toward Gram-positive targets strains. The secondary screening demonstrated an antagonistic activity oriented only against St. aureus ATCC 6538, leading to the selection of five isolates: L_m14, L_m21, L_m23, L_m24, and L_m25 with a larger inhibition zone compared to the others. The ANOVA statistical analysis reveals a small variation of repeatability: L_m21: 0.56%, L_m23: 0%, L_m25: 1.67%, L_m14: 1.88%, L_m24: 2.14%. Conversely, slight variation was reported in terms of inhibition diameters: 9.58± 0.40, 9.83± 0.46 and 10.16± 0.24 8.5 ± 0.40 10 mm for, L_m21, L_m23, L_m25, L_m14and L_m24, indicating that the observed potential showed a heterogeneous distribution (BMS = 0.383, WMS = 0.117). The repeatability coefficient calculated displayed 7.35%. As for the bacteriocins semiquantification, the five samples exhibited production amounts about 4.16 for L_m21, L_m23, L_m25 and 2.08 AU/ml for L_m14, L_m24. Concerning the sensitivity the crude bacteriocins were fully insensitive to heat inactivation, until 121°C, they preserved the same inhibition diameter. As to, kinetic of growth , the µmax showed reductions in pathogens load for L_m21, L_m23, L_m25, L_m14, L_m24 of about 42.92%, 84.12%, 88.55%, 54.95%, 29.97% in the second trails. Inversely, this pathogen growth after five hours displayed differences of 79.45%, 12.64%, 11.82%, 87.88%, 85.66% in the second trails, compared to the control. This study showed potential inhibition to the growth of this food pathogen, suggesting the possibility to improve the hygienic food quality.

Keywords: Exploratory test, lactic acid bacteria, crude bacteriocins, spoilage, pathogens.

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Authors: G. A. Oladosu, P. O. Ogbodogbo, C. I. Makinde, M. O. Tijani, O. A. Adegboyega

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Saprolegniasis is a major pathogenic infection that contributes significantly to poor hatching rates in incubated fish eggs in the African catfish hatchery in Nigeria. Malachite green, known to be very effective against this condition, has been banned because it is carcinogenic. There is therefore the need for other effective, yet safer method of controlling saprolegniasis in incubated fish eggs. A total of 50 ml crude, chloroform extract of pyocyanin from which solvent was removed to attain 30 ml, having a concentration of 12.16 ug/ml was produced from 700 ml broth culture of Pseudomonas aeruginosa isolated from a previous study. In vitro susceptibility of the fungus was investigated by exposing fungal infected eggs to two different time-concentration ratios of pyocyanin; 0.275 ug/ml and 2.75 ug/ml for 1 and 24 h, and 5 mg/L malachite green as positive control while normal saline was the control. Efficacy of pyocyanin was evaluated using the degree of mycelial growth inhibition in the different treatments. Fertilized Clarias gariepinus eggs (between 45 to 64 eggs) were then incubated in 20 ml of medium containing the similar concentrations of pyocyanin and malachite green, with freshwater as control for 24 hours. Hatching rates of the incubated eggs were observed. Three samples of un-hatched eggs were taken from each medium and observed for the presence of fungal pathogens using microscopy. Another batch of three samples of un-hatched eggs from each treatment was also inoculated on Sabourand dextrose agar (SDA) using Egg-Agar Transfer technique to observe for fungal growth. Mycelial growth was inhibited in fungal infected eggs treated with 2.75 ug/ml for 24 h and the 5 mg/L malachite green for both 1 h and 24 h. The mortality rate was 100% in fertilized C. gariepinus eggs exposed for 24 h to 0.275 and 2.75 ug/ml of pyocyanin. The mortality rate was least in the malachite green followed by the control treatment. Embryonic development was observed to be arrested in the eggs treated with the two pyocyanin concentrations as they maintain their color but showed no development beyond the gastrula stage, whereas viable eggs in the control and malachite green treatments developed fully into healthy hatchlings. Furthermore, microscopy of the un-hatched eggs revealed the presence of a protozoan ciliate; Colpidium sp. (Tetrahymenidae), as well as a pathogenic fungus; Saprolegnia sp. in the control, but not in the malachite green and pyocyanin treatments. Growth of Saprolegnia sp. was also observed in SDA culture of un-hatched eggs from the control, but not from pyocyanin and malachite green treated eggs. Pyocyanin treatment of incubated eggs of Clarias gariepinus effectively prevented fungal infection in the eggs, but also arrested the development of the embryo. Therefore, crude chloroform extract of pyocyanin from Pseudomonas aeruginosa cannot be used in the control of Saprolegniasis in incubated Clarias gariepinus eggs at the concentration and duration tested in this study.

Keywords: African catfish, bioprophylaxis, embryo, saprolegniasis.

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Authors: Malkhaz Jokhadze, Vakhtang Mshvildadze, Levan Makaradze, Ekaterine Mosidze, Salome Barbaqadze, Mariam Murtazashvili, Dali Berashvili, Koba sivsivadze, Lasha Bakuridze, Aliosha Bakuridze

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Essential oils are one of the most important groups of biologically active substances present in plants. Due to the chemical diversity of components, essential oils and their preparations have a wide spectrum of pharmacological action. They have bactericidal, antiviral, fungicidal, antiprotozoal, anti-inflammatory, spasmolytic, sedative and other activities. They are expectorant, spasmolytic, sedative, hypotensive, secretion enhancing, antioxidant remedies. Based on preliminary pharmacological studies, we have developed a formulation called “Phytobiotic” containing essential oils, a feed additive for poultry as an alternative to antibiotics. Phytobiotic is a water-soluble powder containing a composition of essential oils of thyme, clary, monarda and auxiliary substances: dry extract of liquorice and inhalation lactose. On this stage of research, the goal was to study the chemical composition of provided phytobiotic, identify the main substances and determine their quantity, investigate the biological activity of phytobiotic through in vitro and in vivo studies. Using gas chromatography-mass spectrometry, 38 components were identified in phytobiotic, representing acyclic-, monocyclic-, bicyclic-, and sesquiterpenes. Together with identification of main active substances, their quantitative content was determined, including acyclic terpene alcohol β-linalool, acyclic terpene ketone linalyl acetate, monocyclic terpenes: D-limonene and γ-terpinene, monocyclic aromatic terpene thymol. Provided phytobiotic has pronounced and at the same time broad spectrum of antibacterial activity. In the cell model, phytobiotic showed weak antioxidant activity, and it was stronger in the ORAC (chemical model) tests. Meanwhile anti-inflammatory activity was also observed. When fowls were supplied feed enriched with phytobiotic, it was observed that gained weight of the chickens in the experimental group exceeded the same data for the control group during the entire period of the experiment. The survival rate of broilers in the experimental group during the growth period was 98% compared to -94% in the control group. As a result of conducted researches probable four different mechanisms which are important for the action of phytobiotics were identified: sensory, metabolic, antioxidant and antibacterial action. General toxic, possible local irritant and allergenic effects of phytobiotic were also investigated. Performed assays proved that formulation is safe.

Keywords: Clary, essential oils, monarda, phytobiotics, poultry, thyme.

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Authors: N. Slepickova Kasalkova, P. Slepicka, L. Bacakova, V. Svorcik

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Tissue engineering includes combination of materials and techniques used for the improvement, repair or replacement of the tissue. Scaffolds, permanent or temporally material, are used as support for the creation of the "new cell structures". For this important component (scaffold), a variety of materials can be used. The advantage of some polymeric materials is their cytocompatibility and possibility of biodegradation. Poly(L-lactic acid) (PLLA) is a biodegradable,  semi-crystalline thermoplastic polymer. PLLA can be fully degraded into H₂O and CO₂. In this experiment, the effect of the surface modification of biodegradable polymer (performed by plasma treatment) on the various cell types was studied. The surface parameters and changes of the physicochemical properties of modified PLLA substrates were studied by different methods. Surface wettability was determined by goniometry, surface morphology and roughness study were performed with atomic force microscopy and chemical composition was determined using photoelectron spectroscopy. The physicochemical properties were studied in relation to cytocompatibility of human osteoblast (MG 63 cells), rat vascular smooth muscle cells (VSMC), and human stem cells (ASC) of the adipose tissue in vitro. A fluorescence microscopy was chosen to study and compare cell-material interaction. Important parameters of the cytocompatibility like adhesion, proliferation, viability, shape, spreading of the cells were evaluated. It was found that the modification leads to the change of the surface wettability depending on the time of modification. Short time of exposition (10-120 s) can reduce the wettability of the aged samples, exposition longer than 150 s causes to increase of contact angle of the aged PLLA. The surface morphology is significantly influenced by duration of modification, too. The plasma treatment involves the formation of the crystallites, whose number increases with increasing time of modification. On the basis of physicochemical properties evaluation, the cells were cultivated on the selected samples. Cell-material interactions are strongly affected by material chemical structure and surface morphology. It was proved that the plasma treatment of PLLA has a positive effect on the adhesion, spreading, homogeneity of distribution and viability of all cultivated cells. This effect was even more apparent for the VSMCs and ASCs which homogeneously covered almost the whole surface of the substrate after 7 days of cultivation. The viability of these cells was high (more than 98% for VSMCs, 89-96% for ASCs). This experiment is one part of the basic research, which aims to easily create scaffolds for tissue engineering with subsequent use of stem cells and their subsequent "reorientation" towards the bone cells or smooth muscle cells.

Keywords: Poly(L-lactic acid), plasma treatment, surface characterization, cytocompatibility, human osteoblasts, rat vascular smooth muscle cells, human stem cells.

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