Search results for: in vitro.

Authors: D. Biswas, Sang H. Hyun

Abstract:

In mammalian reproductive tract, the oviduct secretes huge number of growth factors and cytokines that create an optimal micro-environment for the initial stages of preimplantation embryos. Secretion of these growth factors is stage-specific. Among them, VEGF is a potent mitogen for vascular endothelium and stimulates vascular permeability. Apart from angiogenesis, VEGF in the oviduct may be involved in regulating the oocyte maturation and subsequent developmental process during embryo production in vitro. In experiment 1, to evaluate the effect of VEGF during IVM of porcine COC and subsequent developmental ability after PA and SCNT. The results from these experiments indicated that maturation rates among the different VEGF concentrations were not significant different. In experiment 2, total intracellular GSH concentrations of oocytes matured with VEGF (5-50 ng/ml) were increased significantly compared to a control and VEGF group (500 ng/ml). In experiment 3, the blastocyst formation rates and total cell number per blastocyst after parthenogenesis of oocytes matured with VEGF (5-50 ng/ml) were increased significantly compared to a control and VEGF group (500 ng/ml). Similarly, in experiment 4, the blastocyst formation rate and total cell number per blastocyst after SCNT and IVF of oocytes matured with VEGF (5 ng/ml) were significantly higher than that of oocytes matured without VEGF group. In experiment 5, at 10 hour after the onset of IVF, pronuclear formation rate was evaluated. Monospermy was significantly higher in VEGF-matured oocytes than in the control, and polyspermy and sperm penetration per oocyte were significantly higher in the control group than in the VEGFmatured oocytes. Supplementation with VEGF during IVM significantly improved male pronuclear formation as compared with the control. In experiment 6, type III cortical granule distribution in oocytes was more common in VEGF-matured oocytes than in the control. In conclusion, the present study suggested that supplementation of VEGF during IVM may enhance the developmental potential of porcine in vitro embryos through increase of the intracellular GSH level, higher MPN formation and increased fertilization rate as a consequence of an improved cytoplasmic maturation.

Keywords: angiogenesis, GSH, monospermy, VEGF

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Authors: A. C. B. C. J. Fernandes, V. C. de Jesus, S. Noruziaan, O. F. G. G. Vilela, K. K. Somarin, R. França, F. H. S. L. Pinheiro

Abstract:

Objective: This in vitro study aimed to evaluate the shear bond strength (SBS) of orthodontic tubes after different enamel pre-treatments. Materials and Methods: A total of 39 crown halves were randomly divided into 3 groups (n = 13). Group I (control group) was exposed to prophy paste (PP), 37% phosphoric acid (PA), and a self-etching primer (SEP). Group II received no prophylaxis, but only PA and SEP. Group III was exposed to PP and SEP. The SBS was used to evaluate the bond strength of the orthodontic tubes one year after bonding. One-way ANOVA and Tukey’s post-hoc test were used to compare SBS values between the three groups. The statistical significance was set to 5%. Results: The difference in SBS values of groups I (36.672 ± 9.315 Mpa), II (34.242 ± 9.986 Mpa), and III (39.055 ± 5.565 Mpa) were not statistically significant (P < 0.05). Conclusion: This study suggests that chairside time can be significantly reduced with the use of PP and a SEP without compromising adhesion. Further evidence is needed by means of a split-mouth design trial.

Keywords: Shear bond strength, orthodontic tubes, self-etching primer, pumice, prophy.

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Authors: M. E. Khalifa, A. A. Gobouri

Abstract:

Potential synthesis of a series of 3-amino-4-arylazothiophene derivatives from reaction of 2-cyano-2-phenylthiocarbamoyl acetamide and the appropriate α-halogenated reagents, followed by coupling with different aryl diazonium salts (Japp-Klingemann reaction), and another series of 5-arylazo-thiazol-2-ylcarbamoyl-thiophene derivatives from base-catalyzed intramolecular condensation of 5-arylazo-2-(N-chloroacetyl)amino-thiazole with selected b-keto compounds (Thorpe-Ziegler reaction) was performed. The biological activity of the two series was studied in vitro. Their versatility for pharmaceutical purposes was reported, where they displayed remarkable activities against selected pathogenic microorganisms; Bacillus subtilis, Staphylococcus aureus (Gram positive bacteria), Escherichia coli, Pseudomonas aeruginosa (Gram negative bacteria), and Aspergillus flavus, Candida albicans (fungi) with various degrees related to their chemical structures.

Keywords: 2-Aminothiazole, antimicrobial, monoazo compounds, thiophene, pathogenic microorganisms.

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Authors: E. R. Arakelova, S. G. Grigoryan, F. G. Arsenyan, N. S. Babayan, R. M. Grigoryan, N. K. Sarkisyan

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The nanotechnology offers some exciting possibilities in cancer treatment, including the possibility of destroying tumors with minimal damage to healthy tissue and organs by targeted drug delivery systems. Considerable achievements in investigations aimed at the use of ZnO nanoparticles and nanocontainers in diagnostics and antitumor therapy were described. However, there are substantial obstacles to the purposes to be achieved by the use of zinc oxide nanosize materials in antitumor therapy. Among the serious problems are the techniques of obtaining ZnO nanosize materials. The article presents a new vector delivery system for the known antitumor drug, doxorubicin in the form of polymeric (PEO, starch-NaCMC) hydrogels, in which nanosize ZnO film of a certain thickness are deposited directly on the drug surface on glass substrate by DC-magnetron sputtering of a zinc target. Anticancer activity in vitro and in vivo of those nanosize zinc oxide composites is shown.

Keywords: Anticancer activity, cancer specificity, doxorubicin, zinc oxide.

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Authors: Sopalun K., Thammasiri K., Ishikawa K.

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The effects of chitosan, a biodegradable polymer, were studied in Grammatophyllum speciosum protocorm-like bodies (PLBs) in vitro culture. The chitosan concentration of 0, 5, 10, 15, 20, 25, 50 or 100 mg/l were supplemented in half-strength Murashige and Skoog (1/2 MS) liquid or on agar media containing 2% (w/v) sucrose. The results showed that liquid medium supplemented with 15 mg/l chitosan showed the highest relative growth rate (7-fold increase) of PLBs. On 1/2 MS agar medium supplemented with 25 mg/l chitosan gave the highest relative growth rate (4-fold increase). The relative growth rate of G. speciosum PLBs on agar medium was significantly lower than that in liquid medium. Moreover, chitosan, supplemented to agar medium promoted shoot formation but not rooting. However, supplementation at too high a level, such as 100 mg/l can inhibit growth and kill PLBs.

Keywords: Chitosan, Grammatophyllum speciosum, Growth stimulator

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Authors: Mahmood Ahmad, Asadullah Madni, Muhammad Usman, Abubakar Munir, Naveed Akhtar, Haji M. Shoaib Khan

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This article demonstrated development of controlled release system of an NSAID drug, Diclofenac sodium employing different ratios of Ethyl cellulose. Diclofenac sodium and ethyl cellulose in different proportions were processed by microencapsulation based on phase separation technique to formulate microcapsules. The prepared microcapsules were then compressed into tablets to obtain controlled release oral formulations. In-vitro evaluation was performed by dissolution test of each preparation was conducted in 900 ml of phosphate buffer solution of pH 7.2 maintained at 37 ± 0.5 °C and stirred at 50 rpm. At predetermined time intervals (0, 0.5, 1.0, 1.5, 2, 3, 4, 6, 8, 10, 12, 16, 20 and 24 hrs). The drug concentration in the collected samples was determined by UV spectrophotometer at 276 nm. The physical characteristics of diclofenac sodium microcapsules were according to accepted range. These were off-white, free flowing and spherical in shape. The release profile of diclofenac sodium from microcapsules was found to be directly proportional to the proportion of ethylcellulose and coat thickness. The in-vitro release pattern showed that with ratio of 1:1 and 1:2 (drug: polymer), the percentage release of drug at first hour was 16.91 and 11.52 %, respectively as compared to 1:3 which is only 6.87 % with in this time. The release mechanism followed higuchi model for its release pattern. Tablet Formulation (F2) of present study was found comparable in release profile the marketed brand Phlogin-SR, microcapsules showed an extended release beyond 24 h. Further, a good correlation was found between drug release and proportion of ethylcellulose in the microcapsules. Microencapsulation based on coacervation found as good technique to control release of diclofenac sodium for making the controlled release formulations.

Keywords: Diclofenac sodium, Microencapsulationtechnology, Ethylcellulose, In-Vitro Release Profile

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Authors: Samira Jebahi, Hassane Oudadesse, Eric Wers, Jiheun Elleuch, Hafedh Elfekih, Hassib Keskes, Xuan Vuong Bui, Abdelfatteh Elfeki

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Chitosan (CH) material reinforced by bioactive glass (46S6) was fabricated. 46S6 containing 17% wt% CH was studied in vitro and in vivo. Physicochemical techniques, such as Fourier transform infrared spectroscopy (FT-IR), coupled plasma optical emission spectrometry (ICP-OES) analysis were used. The behavior of 46S6CH17 was studied by measuring the in situ pH in a SBF solution. The 46S6CH17 was implanted in the rat femoral condyl. In vitro 46S6CH17 gave an FTIR - spectrum in which three absorption bands with the maxima at 565, 603 and 1039cm-1 after 3 days of soaking in physiological solution. They are assigned to stretching vibrations of PO4^3- group in phosphate crystalline. Moreover, the pH measurement was decreased in the SBF solution. The stability of the calcium phosphate precipitation depended on the pH value. In vivo, a rise in the Ca and phosphate P ions concentrations in the implanted microenvironment was determined.

Keywords: Bioglass, Chitosan, pH measurement, Hydroxyapatite Carbonateted Layer.

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Authors: M. Shahar Yar, M. Mustaqeem Abdullah, Jaseela Majeed

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A series of 1-(1H-benzimidazol-2-yl)-3-(substituted phenyl)-2-propen-1-one were allowed to react with hydrazine hydrate and phenyl hydrazine in submitted reactions to get pyrazoline and phenyl pyrazoline derivatives. All the compounds entered for screening at the Tuberculosis Antimicrobial Acquisition and Coordinating Facility (TAACF) for their in vitro antibacterial activity against Mycobacterium tuberculosis H37Rv strain (ATCC 27294) using Microplate Alamar Blue Assay (MABA) susceptibility test. The results expressed as MIC (minimum inhibitory concentration) in μg/mL. Among the fifteen compounds, eight compounds were found to have MIC values less than 10 μg/mL. These were subjected for cytotoxicity assay in VERO cells to determine CC50 (cytotoxic concentration 50%) values and finally SI (Selectivity Index) were calculated. Compound (XV) 2-[5-(4- fluorophenyl)-1-phenyl-4,5-dihydro-1H-3-pyrazolyl]-1Hbenzimidazole was considered the best candidate of the series that could be a good starting point to develop new lead compounds in the fight against tuberculosis.

Keywords: anti-tubercular activity, benzimidazole, pyrazoline.

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Authors: F. Bamdad, Sh. Dokhani, J. Keramat, R. Zareie

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Biologically active peptides are of particular interest in food science and human nutrition because they have been shown to play several physiological roles. In vitro gastrointestinal digestion of lentil and whey proteins in this study produced high angiotensin-I converting enzyme inhibitory activity with 75.5±1.9 and 91.4±2.3% inhibition, respectively. High ACE inhibitory activity was observed in lentil after 5 days of germination (84.3±1.2%). Fractionation by reverse phase chromatography gave inhibitory activities as high as 86.3±2.0 for lentil, 94.8±1.8% for whey and 93.7±1.7% at 5th day of germination. Further purification by HPLC resulted in several inhibitory peptides with IC50 values ranging from 0.064 to 0.164 mg/ml. These results demonstrate that lentil proteins are a good source of peptides with ACE inhibitory activity that can be released by germination or gastrointestinal digestion. Despite the lower bioactivity in comparison with whey proteins, incorporation of lentil proteins in functional food formulations and natural drugs look promising.

Keywords: ACE inhibitory peptides, digestion, germination, lentil proteins, whey proteins

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Authors: Chockpisit Thepsithar, Nongnuch Euawong, Nukul Jonghomkajorn

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The in vitro culture procedure of purple nutsedge (Cyperus rotundus L.) for multiple shoot induction and tuber formation was established. Multiple shoots were significantly induced from a single shoot of about 0.5 – 0.8 cm long, on Murashige and Skoog (MS) medium supplemented with 4.44 μM 6- benzyladinine (BA) alone or in combination with 2.85 μM 1- indoleacetic acid (IAA), providing 17.6 and 15.3 shoots per explant with 31.2 and 27.5 leaves per explant, respectively, within 6 weeks of culturing. Moreover, MS medium supplemented with 4.44 μM BA and 2.85 μM IAA was suitable for tuber induction, obtaining 5.9 tubers with 3.4 rhizomes per explant. In combination with ancymidol and higher concentration of sucrose, 11.1 μM BA and 60 g/L sucrose or 11.1 μM BA, 7.8 μM ancymidol and 60 g/L sucrose induced 3.5 tubers with 1.6 rhizomes or 3.5 tubers without rhizome, respectively. However, MS medium containing 3.9 or 7.8 μM ancymidol in combination with either 60 or 80 g/L sucrose enchanced significant root formation at 20.9 – 23.6 roots per explant.

Keywords: Purple nutsedge, Cyperus rotundus, multiple shoot induction, tuber formation

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Authors: N. B. Dhayanithi, T. T. Ajith Kumar, T. Balasubramanian

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Present study summarizes the control of Vibrio alginolyticus infection in hatchery reared Clownfish, Amphiprion sebae with the extract of the mangrove plant, Avicennia marina. Fishes with visible symptoms of hemorrhagic spots were chosen and the genomic DNA of the causative bacterium was isolated and sequenced based on 16S rDNA gene. The in vitro assay revealed that a fraction of A. marina leaf extract elucidated with ethyl acetate: methanol (6:4) showed a high activity (28 mm) at 125 μg/ml concentrations. About 4 % of the fraction fed along with live V. alginolyticus was significantly decreased the cumulative mortality (P<0.05) in the experimental groups than the control group. The responsible fraction was investigated by gas chromatography - mass spectroscopy and found the presence of active compounds. This is the first research in India to control vibriosis infection in marine ornamental fish with mangrove leaf extract.

Keywords: Amphiprion seabe, Avicennia marina, Gas Chromatography - Mass Spectroscopy, Vibrio alginolyticus

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Authors: Marcela Capcarova, Adriana Kolesarova, Marina Medvedova, Peter Petruska, Alexander V. Sirotkin

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The aim of this study was to determine the activity of superoxide dismutase (SOD), glutathione peroxidase (GPx), total antioxidant status (TAS) and accumulation of Hsp70 in porcine ovarian granulosa cells after deoxynivalenol (DON) and zearalenone (ZEA) exposure in vitro. Porcine ovarian granulosa cells were incubated with DON/ZEA administrations as follows: group A (10/10 ng/mL), group B (100/100 ng/mL), group C (1000/1000 ng/mL), and the control group without any additions for 24h. In this study mycotoxins developed stress reaction of porcine ovarian granulosa cells and increased accumulation of Hsp70 what resulted in increasing activities of SOD and GPx in groups with lower doses of mycotoxins. High dose of DON and ZEA had opposite effect on GPx activity than the lower doses. Slight increase in TAS of porcine granulosa cells was observed after mycotoxins exposure. These results contribute towards the understanding of cellular stress and its response.

Keywords: Deoxynivalenol, zearalenone, antioxidants, Hsp70, granulosa cells.

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Authors: Seyed Azizollah Ghotb, Mohammad Chamani, Elmira Abdollahzadeh Esmaeili, Farhad Foroudi

Abstract:

the objective of this study is to measure the levels of cellulas activity of ostrich GI microorganisms, and comparing it with the levels of cellulas activity of rumen-s microorganisms, and also to estimate the probability of increasing enzyme activity with injecting different dosages (30%, 50% and 70%) of pure anaerobic goat rumen fungi. The experiment was conducted in laboratory and under a complete anaerobic condition (in vitro condition). 40 ml of “CaldWell" medium and 1.4g wheat straw were placed in incubator for an hour. The cellulase activity of ostrich microorganisms was compared with other treatments, and then different dosages (30%, 50% and 70%) of pure anaerobic goat rumen fungi were injected to ostrich microorganism-s media. Due to the results, cattle and goat with 2.13 and 2.08 I.U (international units) respectively showed the highest activity and ostrich with 0.91 (I.U) had the lowest cellulose activity (p < 0.05). Injecting 30% and 50% of anaerobic fungi had no significant incensement in enzyme activity, but with injecting 70% of rumen fungi to ostrich microorganisms culture a significant increase was observed 1.48 I.U. (p < 0.05).

Keywords: Cellulase enzyme, Microorganisms, Ostrich, Ruminants

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Authors: R. S. A. Nesbitt, J. Macione, E. Babollah, B. Adu-baffour, S. P. Kotha

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F-actin fibrils are the cytoskeleton of osteocytes. They react in a dynamic manner to mechanical loading, and strength and reposition their efforts to reinforce the cells structure. We hypothesize that f-actin is temporarly disrupted after loading and repolymerizes in a new orientation to oppose the applied load. In vitro studies are conducted to determine f-actin disruption after varying mechanical stimulus parameters that are known to affect bone formation. Results indicate that the f-actin cytoskeleton is disrupted in vitro as a function of applied mechanical stimulus parameters and that the f-actin bundles reassemble after loading induced disruption within 3 minutes after cessation of loading. The disruption of the factin cytoskeleton depends on the magnitude of stretch, the numbers of loading cycles, frequency, the insertion of rest between loading cycles and extracellular calcium. In vivo studies also demonstrate disruption of the f-actin cytoskeleton in cells embedded in the bone matrix immediately after mechanical loading. These studies suggest that adaptation of the f-actin fiber bundles of the cytoskeleton in response to applied loads occurs by disruption and subsequent repolymerization.

Keywords: Mechanical loading of osteocytes, f-actin cytoskeleton, disruption, re-polymerization.

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Authors: Diky Mudhakir, M. Fauzi Bostanudin, Fiki Firmawan, Rachmat Mauludin

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In the present study, development of salbutamol sulphate nanoparticles that adhere to gastric mucus was investigated. Salbutamol sulphate has low bioavailability due to short transit time in gastric. It also has a positive surface charge that provides hurdles to be encapsulated by the positively strong mucoadhesive polymer of chitosan. To overcome the difficulties, the surface charge of active ingredient was modified using several nonionic and anionic stomach-specific polymers. The nanoparticles were prepared using ionotropic gelation technique. The evaluation involved determination of particle size, zeta potential, entrapment efficiency, in vitro drug release and in vitro mucoadhesion test. Results exhibited that the use of anionic alginate polymer was more satisfactory than that of nonionic polymer. Characteristics of the particles was nano-size, high encapsulation efficiency, fulfilled the drug release requirements and adhesive towards stomach for around 11 hours. This result shows that the salbutamol sulphate nanoparticles can be utilized for improvement its delivery.

Keywords: Mucoadhesive, salbutamol sulphate, nanosize, anionic polymer.

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Authors: Laura Elena De León Prado, Dora Alicia Cortés Hernández, Javier Sánchez

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Among the different cancer treatments that are currently used, hyperthermia has a promising potential due to the multiple benefits that are obtained by this technique. In general terms, hyperthermia is a method that takes advantage of the sensitivity of cancer cells to heat, in order to damage or destroy them. Within the different ways of supplying heat to cancer cells and achieve their destruction or damage, the use of magnetic nanoparticles has attracted attention due to the capability of these particles to generate heat under the influence of an external magnetic field. In addition, these nanoparticles have a high surface area and sizes similar or even lower than biological entities, which allow their approaching and interaction with a specific region of interest. The most used magnetic nanoparticles for hyperthermia treatment are those based on iron oxides, mainly magnetite and maghemite, due to their biocompatibility, good magnetic properties and chemical stability. However, in order to fulfill more efficiently the requirements that demand the treatment of magnetic hyperthermia, there have been investigations using ferrites that incorporate different metallic ions, such as Mg, Mn, Co, Ca, Ni, Cu, Li, Gd, etc., in their structure. This paper reports the synthesis of nanosized Mg_xMn_1-xFe₂O₄ (x = 0.3 and 0.4) ferrites by sol-gel method and their evaluation in terms of heating capability and in vitro hemolysis to determine the potential use of these nanoparticles as thermoseeds for the treatment of cancer by magnetic hyperthermia. It was possible to obtain ferrites with nanometric sizes, a single crystalline phase with an inverse spinel structure and a behavior near to that of superparamagnetic materials. Additionally, at concentrations of 10 mg of magnetic material per mL of water, it was possible to reach a temperature of approximately 45°C, which is within the range of temperatures used for the treatment of hyperthermia. The results of the in vitro hemolysis assay showed that, at the concentrations tested, these nanoparticles are non-hemolytic, as their percentage of hemolysis is close to zero. Therefore, these materials can be used as thermoseeds for the treatment of cancer by magnetic hyperthermia.

Keywords: Ferrites, heating capability, hemolysis, nanoparticles, sol-gel.

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Authors: Warachate Khobjai, Suriyan Sukati, Khemjira Jarmkom, Pattaranut Eakwaropas, Surachai Techaoei

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The propose of this study was to investigate in vitro thrombolytic activity of Zingiber cassumunar Roxb. and Prasaplai, a Thai herbal formulation of Z. cassumunar Roxb. Herbs were extracted with boiling water and concentrated by lyophilization. To observe their thrombolytic potential, an in vitro clot lysis method was applied where streptokinase and sterile distilled water were used as positive and negative controls, respectively. Crude aqueous extracts from Z. cassumunar Roxb. and Prasaplai formula showed significant thrombolytic activity by clot lysis of 17.90% and 25.21%, respectively, compared to the negative control water (5.16%) while the standard streptokinase revealed 64.78% clot lysis. These findings suggest that Z. cassumunar Roxb. exhibits moderate thrombolytic activity and cloud play an important role in the thrombolytic properties of Prasaplai formula. However, further study should be done to observe in vivo clot dissolving potential and to isolate active component(s) of these extracts.

Keywords: Aqueous extract, prasaplai formula, thrombolytic activity, Zingiber cassumunar Roxb.

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Authors: Adil Elrayah, Jie Weng, Esra Suliman

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The mineral in human bone is not pure stoichiometric calcium phosphate (Ca/P) as it is partially substituted by in organic elements. In this study, the copper ions (Cu²⁺) substituted hydroxyapatite (CuHA) powder has been synthesized by the co-precipitation method. The CuHA powder has been used to fabricate CuHA fiber scaffolds by sol-gel process and the following sinter process. The resulted CuHA fibers have slightly different microstructure (i.e. porosity) compared to HA fiber scaffold, which is denser. The mechanical properties test was used to evaluate CuHA, and the results showed decreases in both compression strength and hardness tests. Moreover, the in vitro used endothelial cells to evaluate the angiogenesis of CuHA. The result illustrated that the viability of endothelial cell on CuHA fiber scaffold surfaces tends to antigenic behavior. The results obtained with CuHA scaffold give this material benefit in biological applications such as antimicrobial, antitumor, antigens, compacts, filling cavities of the tooth and for the deposition of metal implants anti-tumor, anti-cancer, bone filler, and scaffold.

Keywords: Fiber scaffold, copper ions, hydroxyapatite, hardness, in vitro, mechanical properties.

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Authors: Lina Paola Orozco-Marín, Yuliet Montoya, John Bustamante

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Ischemic events can culminate in acute myocardial infarction with irreversible cardiac lesions that cannot be restored due to the limited regenerative capacity of the heart. Tissue engineering proposes therapeutic alternatives by using biomaterials to resemble the native extracellular medium combined with healthy and functional cells. This research focused on developing a natural thermosensitive hydrogel, its physical-chemical characterization and in vitro biocompatibility determination. Hydrogels’ morphological characterization was carried out through scanning electron microscopy and its chemical characterization by employing Infrared Spectroscopy technic. In addition, the biocompatibility was determined using fetal human ventricular cardiomyocytes cell line RL-14 and the MTT cytotoxicity test according to the ISO 10993-5 standard. Four biocompatible and thermosensitive hydrogels were obtained with a three-dimensional internal structure and two gelation times. The results show the potential of the hydrogel to increase the cell survival rate to the cardiac cell therapies under investigation and lay the foundations to continue with its characterization and biological evaluation both in vitro and in vivo models.

Keywords: cardiac cell therapy, cardiac ischemia, natural polymers, thermosensitive hydrogel

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Authors: Uma Batra, Seema Kapoor

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Commercial hydroxyapatite (HA) was reinforced by adding 2, 5, and 10 wt % of 28.5%CaO-28.5%P2O5-38%Na2 O- 5%CaF2 based glass and then sintered. Although HA shows good biocompatibility with the human body, its applications are limited to non load-bearing areas and coatings due to its poor mechanical properties. These mechanical properties can be improved substantially with addition of glass ceramics by sintering. In this study, the effects of sintering hydroxyapatite with above specified phosphate glass additions are quantified. Each composition was sintered over a range of temperatures. Scanning electron microscopy and x-ray diffraction were used to characterize the microstructure and phases of the composites. The density, microhardness, and compressive strength were measured using Archimedes Principle, Vickers Microhardness Tester (at 0.98 N), and Instron Universal Testing Machine (cross speed of 0.5 mm/min) respectively. These results were used to indicate which composition provided suitable material for use in hard tissue replacement. Composites containing 10 wt % glass additions formed dense HA/TCP (tricalcium phosphate) composite materials possessing good compressive strength and hardness than HA. In-vitro bioactivity was assessed by evaluating changes in pH and Ca2+ ion concentration of SBF-simulated body fluid on immersion of these composites in it for two weeks.

Keywords: Bioglass, Composite, Hydroxyapatite, Sintering.

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Authors: Nguyen Minh Tuan, Yen Chung-Ruey

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By hand pollination was conducted to evaluated different pollen sources and their affects on fruit set and quality of wax apple. The following parameters were recorded: fruit set, seed set, fruit characteristics. Results showed that fruit set percentage with seed were significantly high in ‘Long Red B’ when ‘Black’, ‘Thyto’ were used as pollen parents. Pollen of ‘Black’, ‘Thyto’ resulted in high fruit weight, fruit diameter, fruit length, bigger flesh thickness, better total soluble solids as compared with other pollens. The observation of pollen-growth in vitro revealed that pollen germination at 15% sucrose concentration are required for optimum pollen germination with the high pollen germination were found in ‘Black’, ‘Thyto’. From the result, we concluded that ‘Black’, ‘Thyto’ were proved to be good pollinizers in ‘Long Red B’. Therefore, artificial cross-pollination using ‘Black’, ‘Thyto’ as pollinizers were strongly recommended for ‘Long Red B’ cultivar in wax apple orchard.

Keywords: Wax apple, pollination, pollen source, in vitro, fruit quality.

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Authors: Nagasamy Venkatesh Dhandapani

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The objective of the present study was to develop sustained release oral matrix tablets of anti epileptic drug levetiracetam. The sustained release matrix tablets of levetiracetam were prepared using hydrophilic matrix hydroxypropyl methylcellulose (HPMC) as a release retarding polymer by wet granulation method. Prior to compression, FTIR studies were performed to understand the compatibility between the drug and excipients. The study revealed that there was no chemical interaction between drug and excipients used in the study. The tablets were characterized by physical and chemical parameters and results were found in acceptable limits. In vitro release study was carried out for the tablets using 0.1 N HCl for 2 hours and in phosphate buffer pH 7.4 for remaining time up to 12 hours. The effect of polymer concentration was studied. Different dissolution models were applied to drug release data in order to evaluate release mechanisms and kinetics. The drug release data fit well to zero order kinetics. Drug release mechanism was found as a complex mixture of diffusion, swelling and erosion.

Keywords: Levetiracetam, sustained-release, hydrophilic matrix tablet, HPMC grade K 100 MCR, wet granulation, zero order release kinetics.

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Authors: Hazim J. Al-Daraji

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The aim of this study was to investigate the effects of supplementing the diluent of roosters' semen with different levels of olive oil on motility, viability, morphology and acrosome integrity of chicken spermatozoa after in vitro storage for up to 72 h. Semen was collected from 60 White Layer males (62 wk of age) kept in separated floor pens and randomly divided into six treatment groups (10 males in each group). Experimental groups were as follows: T1 :fresh semen, T2 : semen extended 1:1 with Al – Daraji 2 diluent (AD2D) alone, T3 – T6 :semen samples extended 1:1 with AD2D supplemented with 2 ml, 4 ml, 6 ml or 8 ml of olive oil / 100 ml of diluent, respectively. Semen samples were then stored at 5 °C for 24 h, 48 h or 72 h. There was a clear influence of diluent supplementation with olive oil on the spermatozoa motility profile; olive oil groups (T3, T4, T5 and T6) recorded the highest scores of mass activity and individual motility during all storage periods compared to T1 and T2 groups. In addition, the inclusion of olive oil into semen diluent (T3, T4, T5 and T6) gave significantly higher percentages of viable spermatozoa, normal morphologically spermatozoa and intact acrosomes irrespective of storage period. These results clearly show that supplementation the diluent of roosters' semen with olive oil can improve semen quality when semen samples in vitro stored at 5 °C for up to 72 h.

Keywords: Olive oil, diluent, liquid storage, semen quality of roosters.

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Authors: Seifollah Bahramikia

Abstract:

Amyloid aggregation of polypeptides is related to a growing number of pathologic states known as amyloid disorders. In recent years, blocking or reversing amyloid aggregation via the use of small compounds are considered as two useful approaches in hampering the development of these diseases. In this research, we have compared the ability of several manganese-salen derivatives, as synthetic compounds, and apigenin, as a natural flavonoid, to inhibit of hen egg-white lysozyme (HEWL) aggregation, as an in vitro model system. Different spectroscopic analyses such as Thioflavin T (ThT) and Anilinonaphthalene-8-sulfonic acid (ANS) fluorescence, Congo red (CR) absorbance along with transmission electron microscopy were used in this work to monitor the HEWL aggregation kinetic and inhibition. Our results demonstrated that both type of compounds were capable to prevent the formation of lysozyme amyloid aggregation in vitro. In addition, our data indicated that synthetic compounds had higher activity to inhibit of the β-sheet structures relative to natural compound. Regarding the higher antioxidant activities of the salen derivatives, it can be concluded that in addition to aromatic rings of each of the compounds, the potent antioxidant properties of salen derivatives contributes to lower lysozyme fibril accumulation.

Keywords: Aggregation, anti-amyloidogenic, apigenin, hen egg white lysozyme, salen derivatives.

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Authors: K. Obikeze, P. Mugabo, I. Green, D. Dietrich, A. Burger

Abstract:

Leonotisleonurus a shrub indigenous to Southern Africa is widely used in traditional medicine to treat a variety of conditions ranging from skin diseases and cough to epileptic fits and ‘heart problems’. Studies on the aqueous extract of the leaves have indicated cycloxegenase enzyme inhibitory activity and an antihypertensive effect. Five methanol leaf extract fractions (MLEa - MLEe) of L. leonurus were tested on anaesthetized normotensive male Wistar rats (AWR) and isolated perfused working rat hearts (IWH). Fraction MLEc (0.01mg/kg – 0.05mg/kg) induced significant increases in BP and HR in AWR and positive chronotropic and inotropic effects in IWH (1.0mg/ml – 5.0mg/ml). Pre-administration of atenolol (2.0mg/kg) and prazosin (60μg/kg) significantly inhibited MLEc effect on HR and MAP respectively in vivo, while atenolol (7.0mg/ml) pre-perfusion significantly inhibited MLEc effect in vitro. The hypertensive effect of MLEc is probably via β1agonism. Results also indicate the presence of multiple cardioactive compounds in L. leonurus.

Keywords: Cardiovascular effect, in vitro, in vivo, isolated perfused working heart, Leonotis leonurus, rat.

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Authors: H. Bouaziz, M. Sefi, J. de Lapuente, M. Borras, N. Zeghal

Abstract:

Although, arsenic trioxide has been the subject of toxicological research, in vitro cytotoxicity and genotoxicity studies using relevant cell models and uniform methodology are not well elucidated. Hence, the aim of the present study was to evaluate the cytotoxicity and genotoxicity induced by arsenic trioxide in human keratinocytes (HaCaT) using the MTT [3-(4, 5-dimethylthiazol-2-yl)- 2,5-diphenyltetrazolium bromide] and alkaline single cell gel electrophoresis (Comet) assays, respectively. Human keratinocytes were treated with different doses of arsenic trioxide for 4 h prior to cytogenetic assessment. Data obtained from the MTT assay indicated that arsenic trioxide significantly reduced the viability of HaCaT cells in a dose-dependent manner, showing an IC50 value of 34.18 ± 0.6 μM. Data generated from the comet assay also indicated a significant dose-dependent increase in DNA damage in HaCaT cells associated with arsenic trioxide exposure. We observed a significant increase in comet tail length and tail moment, showing an evidence of arsenic trioxide -induced genotoxic damage in HaCaT cells. This study confirms that the comet assay is a sensitive and effective method to detect DNA damage caused by arsenic.

Keywords: Arsenic trioxide, cytotoxixity, genotoxicity, HaCaT.

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Authors: Nikoo Soleimani

Abstract:

Background: Reducing torque during bone drilling is one of the effective factors in reaching to an optimal drilling process. Methods: 15 bovine femurs were drilled in vitro with a drill bit with a diameter of 4 mm using two methods of torsional ultrasonic assisted drilling (T-UAD) and convent conventional drilling (CD) and the effects of changing the feed rate and rotational speed on the torque were compared in both methods. Results: There was no significant difference in the thrust force measured in both methods due to the direction of vibrations. Results showed that using T-UAD method for bone drilling at feed rates of 0.16, 0.24 and 0.32 mm/rev led for all rotational speeds to a decrease of at least 16.3% in torque compared to the CD method. Further, using T-UAD at rotational speeds of 355~1000 rpm with various feed rates resulted in a torque reduction of 16.3~50.5% compared to CD method. Conclusions: Reducing the feed rate and increasing the rotational speed, except for the rotational speed of 500 rpm and a feed rate of 0.32 mm/rev, resulted generally in torque reduction in both methods. However, T-UAD is a more effective and desirable option for bone drilling considering its significant torque reduction.

Keywords: Torsional ultrasonic assisted drilling, torque, bone drilling, rotational speed, feed rate.

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Authors: Ihor Trots, Yuriy Tasinkevych, Andrzej Nowicki, Marcin Lewandowski

Abstract:

In the paper the study of synthetic transmit aperture method applying the Golay coded transmission for medical ultrasound imaging is presented. Longer coded excitation allows to increase the total energy of the transmitted signal without increasing the peak pressure. Moreover signal-to-noise ratio and penetration depth are improved while maintaining high ultrasound image resolution. In the work the 128-element linear transducer array with 0.3 mm inter-element spacing excited by one cycle and the 8 and 16- bit Golay coded sequences at nominal frequency 4 MHz was used. To generate a spherical wave covering the full image region a single element transmission aperture was used and all the elements received the echo signals. The comparison of 2D ultrasound images of the tissue mimicking phantom and in vitro measurements of the beef liver is presented to illustrate the benefits of the coded transmission. The results were obtained using the synthetic aperture algorithm with transmit and receive signals correction based on a single element directivity function.

Keywords: Golay coded sequences, radiation pattern, signal processing, synthetic aperture, ultrasound imaging.

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Authors: Ayu S. Muhamad, Michael Gleeson

Abstract:

Immunomodulators are substances that alter immune system via dynamic regulation of messenger molecules. It can be divided into immunostimulant and immunosuppressant. It can help to increase immunity of people with a low immune system, and also can help to normalize an overactive immune system. Aim of this study is to investigate the effects of in vitro exposure to low and high doses of several immunomodulators which include caffeine, kaloba and quercetin on antigen-stimulated whole blood culture cytokine production. Whole blood samples were taken from 5 healthy males (age: 32 ± 12 years; weight: 75.7 ± 6.1 kg; BMI: 24.3 ± 1.5 kg/m2) following an overnight fast with no vigorous activity during the preceding 24 h. The whole blood was then stimulated with 50 μl of 100 x diluted Pediacel vaccine and low or high dose of immunomodulators in the culture plate. After 20 h incubation (5% CO2, 37°C), it was analysed using the Evidence Investigator to determine the production of cytokines including IL-2, IL-4, IL-10, IFN-γ, and IL-1α. Caffeine and quercetin showed a tendency towards decrease cytokine production as the doses were increased. On the other hand, an upward trend was evident with kaloba, where a high dose of kaloba seemed to increase the cytokine production. In conclusion, we found that caffeine and quercetin have potential as immunosuppressant and kaloba as immunostimulant.

Keywords: Caffeine, cytokine, immunomodulators, kaloba, quercetin.

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Authors: Imran Khan Pathan, Anil Bhandari, Peeyush K. Sharma, Rakesh K. Patel, Suresh Purohit

Abstract:

Floating tablets of Marichyadi Vati were developed with an aim to prolong its gastric residence time and increase the bioavailability of drug. Rapid gastrointestinal transit could result in incomplete drug release from the drug delivery system above the absorption zone leading to diminished efficacy of the administered dose. The tablets were prepared by wet granulation technique, using HPMC E50 LV act as Matrixing agent, Carbopol as floating enhancer, microcrystalline cellulose as binder, Sodium bi carbonate as effervescent agent with other excipients. The simplex lattice design was used for selection of variables for tablets formulation. Formulation was optimized on the basis of floating time and in vitro drug release. The results showed that the floating lag time for optimized formulation was found to be 61 second with about 97.32 % of total drug release within 3 hours. The vitro release profiles of drug from the formulation could be best expressed zero order with highest linearity r² = 0.9943. It was concluded that the gastroretentive drug delivery system can be developed for Marichyadi Vati containing Piperine to increase the residence time of the drug in the stomach and thereby increasing bioavailability.

Keywords: Piperine, Marichyadi Vati, Gastroretentive drug delivery, Floating tablet.

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