Search results for: in vitro study

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: H. Feza Carlak, N. G. Gencer

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To increase the temperature contrast in thermal images, the characteristics of the electrical conductivity and thermal imaging modalities can be combined. In this experimental study, it is objected to observe whether the temperature contrast created by the tumor tissue can be improved just due to the current application within medical safety limits. Various thermal breast phantoms are developed to simulate the female breast tissue. In vitro experiments are implemented using a thermal infrared camera in a controlled manner. Since experiments are implemented in vitro, there is no metabolic heat generation and blood perfusion. Only the effects and results of the electrical stimulation are investigated. Experimental study is implemented with two-dimensional models. Temperature contrasts due to the tumor tissues are obtained. Cancerous tissue is determined using the difference and ratio of healthy and tumor images. 1 cm diameter single tumor tissue causes almost 40 °mC temperature contrast on the thermal-breast phantom. Electrode artifacts are reduced by taking the difference and ratio of background (healthy) and tumor images. Ratio of healthy and tumor images show that temperature contrast is increased by the current application.

Keywords: Medical diagnostic imaging, breast phantom, active thermography, breast cancer detection.

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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: Iriawati, R. R. Esyanti, W. Natalia, N. Zahya

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In vitro plant regeneration has been successfully obtained from basal shoot explant of Vetiveria zizanioides through indirect organogenesis. The explant was cultured in Murashige & Skoog’s (MS) media supplemented with 2,4-D, IAA, and kinetin in various concentrations. Callus was well induced in media supplemented with 2 ppm 2,4-D, 1 ppm IAA, and 1 ppm kinetin. This callus was then transferred to MS media supplemented with 1 - 5 ppm of BAP for shoot regeneration. The media supplemented with 3 ppm BAP was a suitable medium for shoot induction, as well as for shoot multiplication. Rooting was well developed in shoot following transferred to half MS media containing 0.2 ppm IBA. Plantlet was then transferred to husk charcoal for acclimatization, and almost all (90%) of plantlets were survived during acclimatization.

Keywords: Callus, plantlet regeneration, shoot induction, Vetiveria zizanioides.

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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: D. Biswas, Sang H. Hyun

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

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

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

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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: 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: K. Biswas, U. H. Armin, S. M. J. Prodhan, J. A. Prithul, S. Sarker, F. Afrin

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Alzheimer’s disease (AD) (a progressive neurodegenerative disorder) is mostly predominant cause of dementia in the elderly. Prolonging the function of acetylcholine by inhibiting both acetylcholinesterase and butyrylcholinesterase is most effective treatment therapy of AD. Traditionally Pterocarpus santalinus L. is widely known for its medicinal use. In this study, in vitro acetylcholinesterase inhibitory activity was investigated and methanolic extract of the plant showed significant activity. To confirm this activity (in vivo), learning and memory enhancing effects were tested in mice. For the test, memory impairment was induced by scopolamine (cholinergic muscarinic receptor antagonist). Anti-amnesic effect of the extract was investigated by the passive avoidance task in mice. The study also includes brain acetylcholinesterase activity. Results proved that scopolamine induced cognitive dysfunction was significantly decreased by administration of the extract solution, in the passive avoidance task and inhibited brain acetylcholinesterase activity. These results suggest that bark extract of Pterocarpus santalinus can be better option for further studies on AD via their acetylcholinesterase inhibitory actions.

Keywords: Pterocarpus santalinus, cholinesterase inhibitor, passive avoidance, Alzheimer’s disease.

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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: 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: 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: Nikoo Soleimani

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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: Mee-Lin Lim Chai Teo, Darryl M. Small

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Wheat has a bimodal starch granule population and the dependency of the rate of enzymatic hydrolysis on particle size has been investigated. Ungelatinised wheaten starch granules were separated into two populations by sedimentation and decantation. Particle size was analysed by laser diffraction and morphological characteristics were viewed using SEM. The sedimentation technique though lengthy, gave satisfactory separation of the granules. Samples (<10μm, >10μm and original) were digested with a-amylase using a dialysis model. Granules of <10μm showed significantly higher rate of reducing sugar release than those >10μm (p<0.05). In contrast, the rate was not significantly different between the original sample and granules >10μm. Moreover, the digestion rate was dependent on particle size whereby smaller granules produced higher rate of release. The methodology and results reported here can be used as a basis for further evaluations designed to delay the release of glucose during the digestion of native starches.

Keywords: in vitro Digestion, a-amylase, wheat starch, granule size.

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Authors: Nélio Veiga, Paula Ferreira, Tiago Correia, Maria J. Correia, Carlos Pereira, Odete Amaral, Ilídio J. Correia

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Introduction: The application of fissure sealants is considered to be an important primary prevention method used in dental medicine. However, the formation of microleakage gaps between tooth enamel and the fissure sealant applied is one of the most common reasons of dental caries development in teeth with fissure sealants. The association between various dental biomaterials may limit the major disadvantages and limitations of biomaterials functioning in a complementary manner. The present study consists in the incorporation of a cariostatic agent – silver diamine fluoride (SDF) – in a resin-based fissure sealant followed by the study of release kinetics by spectrophotometry analysis of the association between both biomaterials and assessment of the inhibitory effect on the growth of the reference bacterial strain Streptococcus mutans (S. mutans) in an in vitro study. Materials and Methods: An experimental in vitro study was designed consisting in the entrapment of SDF (Cariestop^® 12% and 30%) into a commercially available fissure sealant (Fissurit^®), by photopolymerization and photocrosslinking. The same sealant, without SDF was used as a negative control. The effect of the sealants on the growth of S. mutans was determined by the presence of bacterial inhibitory halos in the cultures at the end of the incubation period. In order to confirm the absence of bacteria in the surface of the materials, Scanning Electron Microscopy (SEM) characterization was performed. Also, to analyze the release profile of SDF along time, spectrophotometry technique was applied. Results: The obtained results indicate that the association of SDF to a resin-based fissure sealant may be able to increase the inhibition of S. mutans growth. However, no SDF release was noticed during the in vitro release studies and no statistical significant difference was verified when comparing the inhibitory halo sizes obtained for test and control group.  Conclusions: In this study, the entrapment of SDF in the resin-based fissure sealant did not potentiate the antibacterial effect of the fissure sealant or avoid the immediate development of dental caries. The development of more laboratorial research and, afterwards, long-term clinical data are necessary in order to verify if this association between these biomaterials is effective and can be considered for being used in oral health management. Also, other methodologies for associating cariostatic agents and sealant should be addressed.

Keywords: Biomaterial, fissure sealant, primary prevention, silver diamine fluoride, S. mutans.

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Authors: Mala Nath, Nagamani Kompelli, Partha Roy, Snehasish Das

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Two new metal-based anticancer chemotherapeutic agents, [(Ph2Sn)2(HGuO)2(phen)Cl2] 1 and [(Ph3Sn)(HGuO)(phen)]- Cl.CH3OH.H2O 2, were designed, prepared and characterized by analytical and spectral (IR, ESI-Mass, 1H, 13C and 119Sn NMR) techniques. The proposed geometry of Sn(IV) in 1 and 2 is distorted octahedral and distorted trigonal-bipyramidal, respectively. Both 1 and 2 exhibit potential cytotoxicity in vitro against MCF-7, HepG-2 and DU-145 cell lines. The intrinsic binding constant (Kb) values of 1 (2.33 × 105 M-1) and 2 (2.46 × 105 M-1) evaluated from UV-Visible absorption studies suggest non-classical electrostatic mode of interaction via phosphate backbone of DNA double helix. The Stern- Volmer quenching constant (Ksv) of 1 (9.74 × 105 M-1) and 2 (2.9 × 106 M-1) determined by fluorescence studies suggests the groove binding and intercalation mode for 1 and 2, respectively. Effective cleavage of pBR322 DNA is induced by 1.Their interaction with DNA of cancer cells may account for potency.

Keywords: Anticancer agents, DNA binding studies, NMR spectroscopy, organotin.

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Authors: Ayu S. Muhamad, Michael Gleeson

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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: P. Massanyi, L. Kichi, T. Slanina, E. Kolesar, J. Danko, N. Lukac, E. Tvrda, R. Stawarz, A. Kolesarova

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Target of this study was the analysis of the impact of crude glycerol on canine spermatozoa motility, morphology, viability, and membrane integrity. Experiments were realized in vitro. In the study, semen from 5 large dog breeds was used. They were typical representatives of large breeds, coming from healthy rearing, regularly vaccinated and integrated to the further breeding. Semen collections were realized at the owners of animals and in the veterinary clinic. Subsequently the experiments were realized at the Department of Animal Physiology of the SUA in Nitra. The spermatozoa motility was evaluated using CASA analyzer (SpermVisionTM, Minitub, Germany) at the temperature 5 and 37°C for 5 hours. In the study, 13 motility parameters were evaluated. Generally, crude glycerol has generally negative effect on spermatozoa motility. Morphological analysis was realized using Hancock staining and the preparations were evaluated at magnification 1000x using classification tables of morphologically changed spermatozoa. Data clearly detected the highest number of morphologically changed spermatozoa in the experimental groups (know twisted tails, tail torso and tail coiling). For acrosome alterations swelled acrosomes, removed acrosomes and acrosomes with undulated membrane were detected. In this study also the effect of crude glycerol on spermatozoa membrane integrity were analyzed. The highest crude glycerol concentration significantly affects spermatozoa integrity. Results of this study show that crude glycerol has effect of spermatozoa motility, viability, and membrane integrity. Detected changes are related to crude glycerol concentration, temperature, as well as time of incubation.

Keywords: Dog, semen, spermatozoa, acrosome, glycerol, CASA, viability.

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Authors: T. Azouaoui – Ait Kettout, R. Gaceb – Terrak, F. Rahmania

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Date production in North Africa is facing a worrying slowdown and a decline because of Fusarium wilt or bayoud date palm (Phoenix dactylifera L., caused by Fusarium oxysporum f. sp. albedinis (F. o. a). The objective of this work is to study the in vitro effect of flavonoid aglycones extracted from the roots of two cultivars of date palm (one sensitive to bayoud (Deglet Nour) and the other resistant (Takerboucht)) on the growth and production fusaric acid of the pathogen. Results show that during the first week of development of F. o. a on potato dextrose liquid medium, the flavonoid aglycones extracts of the susceptible cultivar roots stimulates mycelial growth as well as conidiogenesis of F.o.a, nevertheless it has no effect on the synthesis of fusaric acid. However, the flavonoid aglycones extract of resistant cultivar roots stimulates mycelial growth and decreases both the number of conidia production and fusaric acid. It therefore appears possible that the resistant cultivar aglycones have two types of action: they either inhibit the synthesis of fusaric acid, or they metabolize this toxin into hydrosoluble product, this is called detoxification.

Keywords: Flavonoid Aglycones, date palm, fusaric acid, Fusarium oxysporum f. sp. albedinis.

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

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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

Abstract:

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.

Abstract:

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

Abstract:

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: Eva Tvrdá, Hana Greifová, Peter Ivanič, Norbert Lukáč

Abstract:

The aim of this study was to evaluate the dose- and time-dependent in vitro effects of berberine (BER), a natural alkaloid with numerous biological properties on bovine spermatozoa during three time periods (0 h, 2 h, 24 h). Bovine semen samples were diluted and cultivated in physiological saline solution containing 0.5% DMSO together with 200, 100, 50, 10, 5, and 1 μmol/L BER. Spermatozoa motility was assessed using the computer assisted semen analyzer. The viability of spermatozoa was assessed by the metabolic (MTT) assay, production of superoxide radicals was quantified using the nitroblue tetrazolium (NBT) test, and chemiluminescence was used to evaluate the generation of reactive oxygen species (ROS). Cell lysates were prepared and the extent of lipid peroxidation (LPO) was evaluated using the TBARS assay. The results of the movement activity showed a significant increase in the motility during long term cultivation in case of concentrations ranging between 1 and 10 μmol/L BER (P < 0.01; P < 0.001; 24 h). At the same time, supplementation of 1, 5 and 10 μmol/L BER led to a significant preservation of the cell viability (P < 0.001; 24 h). BER addition at a range of 1-50 μmol/L also provided a significantly higher protection against superoxide (P < 0.05) and ROS (P < 0.001; P < 0.01) overgeneration as well as LPO (P < 0.01; P<0.05) after a 24 h cultivation. We may suggest that supplementation of BER to bovine spermatozoa, particularly at concentrations ranging between 1 and 50 μmol/L, may offer protection to the motility, viability and oxidative status of the spermatozoa, particularly notable at 24 h.

Keywords: Berberine, bulls, motility, oxidative profile, spermatozoa, viability.

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

Abstract:

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: Chockpisit Thepsithar, Nongnuch Euawong, Nukul Jonghomkajorn

Abstract:

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: Zeinab Hajifathali, Moghan Amirhosseinian

Abstract:

This study had two main aims: firstly, to determine how the individual substitution of CaO/MgO and CaO/SrO can affect the in vitro bioactivity of sol-gel derived substituted 58S bioactive glass (BG) and secondly to introduce a composition in the 60SiO₂–(36-x)CaO–4P₂O₅–(x)MgO and 60SiO₂–(36-x)CaO–4P₂O₅–(x)SrO quaternary systems (where x= 0, 5, 10 mol.%) with enhanced biocompatibility, alkaline phosphatase (ALP) activity, and more efficient antibacterial activity against MRSA bacteria. Results showed that both magnesium-substituted bioactive glasses (M-BGs) and strontium- substituted bioactive glasses (S-BGs) retarded the Hydroxyapatite (HA) formation. Meanwhile, magnesium had more pronounced effect. The 3-(4, 5dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and ALP assays revealed that the presence of moderate amount (5 mol%) of Mg and Sr had a stimulating effect on increasing of both proliferation and differentiation of MC3T3-E1 cells. Live dead and Dapi/actin staining revealed both substitution of CaO/MgO and CaO/SrO resulted in more biocompatibility and stimulation potential of the MC3T3 cells compared with control. Taken together, among all of the synthesized magnesium substituted (MBGs) and strontium substituted (SBGs), the sample 58- BG with 5 mol% CaO/MgO substitution (BG-5M) was considered as a multifunctional biomaterial in bone tissue regeneration field with enhanced biocompatibility, ALP activity as well as the highest antibacterial efficiency against methicillin-resistant Staphylococcus aureus (MRSA) bacteria.

Keywords: Apatite, alkaline earth, bioactivity, biomedical applications, sol-gel.

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