Formulation and in vitro Evaluation of Ondansetron Hydrochloride Matrix Transdermal Systems Using Ethyl Cellulose/Polyvinyl Pyrrolidone Polymer Blends

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.

Chemical and Biological Properties of Local Cowpea Seed Protein Grown in Gizan Region

The aim of the present study was to investigate the chemical and biological properties of local cowpea seed protein cultivated in Gizan region. The results showed that the cowpea and its products contain high level of protein (22.9-77.6%), high carbohydrates (9.4-64.3%) and low fats (0.1-0.3%). The trypsin and chymotrypsin activities were found to be 32.2 and 15.2 units, respectively. These activities were not affected in both defatted and protein concentrate whereas they were significantly reduced in isolated protein and cooked samples. The phytate content of cooked and concentrated cowpea samples varied from 0.25% -0.32%, respectively. Tannin content was found to be 0.4% and 0.23% for cooked and raw samples, respectively. The in vitro protein digestibility was very high in cowpea seeds (75.04-78.76%). The biological evaluation using rats showed that the group fed with animal feed containing casein gain more weight than those fed with that containing cowpea. However, the group fed with cooked cowpea gain more weight than those fed with uncooked cowpea. On the other hand, in vivo digestion showed high value (98.33%) among the group consumed casein compared to other groups those consumed cowpea contains feed. This could be attributed to low antinutritional factors in casein contains feed compared to those of cowpea contains feed because cooking significantly increased the digestion rate (80.8% to 83.5%) of cowpea contains feed. Furthermore, the biological evaluation was high (91.67%) of casein containing feed compared to that of cowpea containing feed (80.83%-87.5%). The net protein utilization (NPU) was higher (89.67%) in the group fed with casein containing feed than that of cowpea containing feed (56.33%-69.67%).

The Effect of Granule Size on the Digestibility of Wheat Starch Using an in vitro Model

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 and original) were digested with a-amylase using a dialysis model. Granules of 10μm (p10μm. 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.

Cultivation of Thymus by In Vitro And Hydroponics Combined Method

Our results showed that for the growth of qualitative seedling and vegetative raw material of ðó. marschallianus Willd. and T. serphyllum L. it is more profitable to use the in vitro and hydroponics combined method. In in vitro culture it is possible to do micro-propagation whole year with 98-99% rhizogenesis. 30000 micro-plants were obtained from one explant during 9 months. Hydroponic conditions provide the necessary microclimate for microplants where the survival rate without acclimatization was 93.3%. The essential oil content in hydroponic dry herb of both species in vegetative and blossom phase was 1.3% whereas in wild plants it was 1.2%, the content of extractive substances and vitamin C also exceeded wild plants. Our biochemical and radiochemical investigations indicated that the medicinal raw materials obtained from hydroponic and wild plants of Thymus species correspond to the demands of SPh XI, and the content of artificial radionuclides does not exceed the MACL.

The Effect of Carboxymethyl Cellulose on the Stability of Emulsions Stabilized by Whey Proteins under Digestion in vitro and in vivo

In vitro gastro-duodenal digestion model was used to investigate the changes of emulsions under digestion conditions. Oil in water emulsions stabilized by whey proteins (2%) and stabilized by whey proteins (2%) with addition of carboxymethyl cellulose (0.75%) as gelling agent of continuous phase were prepared at pH7. Both emulsions were destabilized under gastric conditions; however the protective role of carboxymethyl cellulose was indicated by recording delay of fat digestibility of this emulsion. In the presence of carboxymethyl cellulose whey proteins on the interfacial surface of droplets were more resistant to gastric degradation causing limited hydrolysis of fat due to the poor acceptability of lipids for the enzymes. Studies of emulsions using in vivo model supported results from in vitro studies. Lower content of triglycerides in blood serum and higher amount of fecal fat of rats were determined when rats were fed by diet containing emulsion made with whey proteins and carboxymethyl cellulose. 

Compressive Properties of a Synthetic Bone Substitute for Vertebral Cancellous Bone

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

Inhibition Effect of Brazilin to Human Bladder Cancer Cell Line T24

The inhibition effect of brazilin to human bladder tumor cell line T24 in vitro and in vivo was studied. The results of the in vitro experiments showed that brazilin has strong inhibition activity on the target cells. The inhibition ratio of 100 μg/mL brazilin and 100 μg/mL mitomycin to the target cells was 90.90 % and 63.24 % respectively, which showed that brazilin has higher inhibition activity than mitomycin under the same concentration. Brazilin could induce cell apoptosis in T24 cells. Significant antitumor activity of brazilin was also showed in the animals experiments. The life extention rate of 200 mg/mL, 300 mg/kg, and 400 mg/kg brazilin intraperitoneally injected into Balb/c-nu-nu nude mice that with human bladder cancer were 51.50 %, 56.90 %, and 58.42 %(P

The Influence of Some Polyphenols on Human Erythrocytes Glutathione S-Transferase Activity

Glutathione S-transferase was purified from human erythrocytes and effects of some polyphenols were investigated on the enzyme activity. The purification procedure was performed on Glutathione-Agarose affinity chromatography after preparation of erythrocytes hemolysate with a yield of 81%. The purified enzyme showed a single band on the SDS-PAGE. The effects of some poliphenolic compounds such as catechin, dopa, dopamine, progallol and catechol were examined on the in vitro GST activity. Catechin was determined to be inhibitor for the enzyme, but others were not effective on the enzyme as inhibitors or activators. IC50 value -the concentration of inhibitor which reduces enzyme activity by 50%- was estimated to be 10 mM. Ki constants were also calculated as 6.38 ± 0,70 mM with GSH substrate, and 3.86 ± 0,78 mM with CDNB substrate using the equations of graphs for the inhibitor, and its inhibition type was determined as non-competitive.

In vitro Studies of Mucoadhesiveness and Release of Nicotinamide Oral Gels Prepared from Bioadhesive Polymers

The aim of the present study was to evaluate the mucoadhesion and the release of nicotinamide gel formulations using in vitro methods. An agar plate technique was used to investigate the adhesiveness of the gels whereas a diffusion apparatus was employed to determine the release of nicotinamide from the gels. In this respect, 10% w/w nicotinamide gels containing bioadhesive polymers: Carbopol 934P (0.5-2% w/w), hydroxypropylmethyl cellulose (HPMC) (4-10% w/w), sodium carboxymethyl cellulose (SCMC) (4-6% w/w) and methylcellulose 4000 (MC) (3-5% w/w) were prepared. The gel formulations had pH values in the range of 7.14 - 8.17, which were considered appropriate to oral mucosa application. In general, the rank order of pH values appeared to be SCMC > MC4000 > HPMC > Carbopol 934P. Types and concentrations of polymers used somewhat affected the adhesiveness. It was found that anionic polymers (Carbopol 934 and SCMC) adhered more firmly to the agar plate than the neutral polymers (HPMC and MC 4000). The formulation containing 0.5% Carbopol 934P (F1) showed the highest release rate. With the exception of the formulation F1, the neutral polymers tended to give higher relate rates than the anionic polymers. For oral tissue treatment, the optimum has to be balanced between the residence time (adhesiveness) of the formulations and the release rate of the drug. The formulations containing the anionic polymers: Carbopol 934P or SCMC possessed suitable physical properties (appearance, pH and viscosity). In addition, for anionic polymer formulations, justifiable mucoadhesive properties and reasonable release rates of nicotinamide were achieved. Accordingly, these gel formulations may be applied for the treatment of oral mucosal lesions.

Magnesium Alloy: A Biomaterial for Development of Degradation Rate Controllable Esophageal Stent

Magnesium alloy has been widely investigated as biodegradable cardiovascular stent and bone implant. Its application for biodegradable esophageal stenting remains unexplored. This paper reports the biodegradation behaviors of AZ31 magnesium alloy in artificial saliva and various types of beverage in vitro. Results show that the magnesium ion release rate of AZ31 in artificial saliva for a stent (2cm diameter, 10cm length at 50% stent surface coverage) is 43 times lower than the daily allowance of human body magnesium intakes. The degradation rates of AZ31 in different beverages could also be significantly different. These results suggest that the esophagus in nature is a less aggressive chemical environment for degradation of magnesium alloys. The significant difference in degradation rates of AZ31 in different beverages opens new opportunities for development of degradation controllable esophageal stent through customizing ingested beverages.

Some Peculiarities of Growth and Functional Activity of Escherichia coli Strain from Probiotic Formula “ASAP“

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

Neurogenic Potential of Clitoria ternatea Aqueous Root Extract–A Basis for Enhancing Learning and Memory

The neurogenic potential of many herbal extracts used in Indian medicine is hitherto unknown. Extracts derived from Clitoria ternatea Linn have been used in Indian Ayurvedic system of medicine as an ingredient of “Medhya rasayana", consumed for improving memory and longevity in humans and also in treatment of various neurological disorders. Our earlier experimental studies with oral intubation of Clitoria ternatea aqueous root extract (CTR) had shown significant enhancement of learning and memory in postnatal and young adult Wistar rats. The present study was designed to elucidate the in vitro effects of 200ng/ml of CTR on proliferation, differentiation and growth of anterior subventricular zone neural stem cells (aSVZ NSC-s) derived from prenatal and postnatal rat pups. Results show significant increase in proliferation and growth of neurospheres and increase in the yield of differentiated neurons of aSVZ neural precursor cells (aSVZNPC-s) at 7 days in vitro when treated with 200ng/ml of CTR as compared to age matched control. Results indicate that CTR has growth promoting neurogenic effect on aSVZ neural stem cells and their survival similar to neurotrophic factors like Survivin, Neuregulin 1, FGF-2, BDNF possibly the basis for enhanced learning and memory.

Design of Salbutamol Sulphate Gastroretentive Nanoparticles via Surface Charge Manipulation

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.

Phosphorus Supplementation of Ammoniated Rice Straw on Rumen Fermentability, Syntesised Microbial Protein and Degradabilityin Vitro

The effect of phosphorus supplementation of ammoniated rice straw was studied. The in vitro experiment was carried out following the first stage of Tilley and Terry method. The treatments consisting of four diets were A = 50% ammoniated rice straw + 50% concentrate (control), B = A + 0.2% Phosphor (P) supplement, C = A + 0.4% Phosphor (P) supplement, and D = A + 0.6% Phosphor (P) supplement of dry matter. Completely randomized design was used as the experimental design with differences among treatment means were examined using Duncan multiple range test. Variables measured were total bacterial and cellulolytic bacterial population, cellulolytic enzyme activity, ammonia (NH3) and volatile fatty acid (VFA) concentrations, as fermentability indicators and synthesized microbial protein, as well as degradability indicators including dry matter (DM), organic matter (OM), neutral detergent fibre (NDF), acid detergent fibre (ADF) and cellulose. The results indicated that fermentability and degradability of diets consisting ammoniated rice straw with P supplementation were significantly higher than the control diet (P< 0.05). It is concluded that P supplementation is important to improve fermentability and degradability of rations containing ammoniated RS and concentrate. In terms of the most effective level of P supplementation occurred at a supplementation rate of 0.4% of dry matter.

Screening and Evaluation of in vivo and in vitro Generated Insulin Plant (Vernonia divergens) for Antimicrobial and Anticancer Activities

Vernonia divergens Benth., commonly known as “Insulin Plant” (Fam: Asteraceae) is a potent sugar killer. Locally the leaves of the plant, boiled in water are successfully administered to a large number of diabetic patients. The present study evaluates the putative anti-diabetic ingredients, isolated from the in vivo and in vitro grown plantlets of V. divergens for their antimicrobial and anticancer activities. Sterilized explants of nodal segments were cultured on MS (Musashige and Skoog, 1962) medium in presence of different combinations of hormones. Multiple shoots along with bunch of roots were regenerated at 1mg l-1 BAP and 0.5 mg l-1 NAA. Micro-plantlets were separated and sub-cultured on the double strength (2X) of the above combination of hormones leading to increased length of roots and shoots. These plantlets were successfully transferred to soil and survived well in nature. The ethanol extract of plantlets from both in vivo & in vitro sources were prepared in soxhlet extractor and then concentrated to dryness under reduced pressure in rotary evaporator. Thus obtainedconcentrated extracts showed significant inhibitory activity against gram negative bacteria like Escherichia coli and Pseudomonas aeruginosa but no inhibition was found against gram positive bacteria. Further, these ethanol extracts were screened for in vitro percentage cytotoxicity at different time periods (24 h, 48 h and 72 h) of different dilutions. The in vivo plant extract inhibited the growth of EAC mouse cell lines in the range of 65, 66, 78, and 88% at 100, 50, 25 & 12.5μg mL-1 but at 72 h of treatment. In case of the extract of in vitro origin, the inhibition was found against EAC cell lines even at 48h. During spectrophotometric scanning, the extracts exhibited different maxima (ʎ) - four peaks in in vitro extracts as against single in in vivo preparation suggesting the possible change in the nature of ingredients during micropropagation through tissue culture techniques.

Interactions between Cells and Nanoscale Surfaces of Oxidized Silicon Substrates

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

3D Locomotion and Fractal Analysis of Goldfish for Acute Toxicity Bioassay

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

Effect of Recombinant Human Follicle Stimulating Hormone on Meiotic Competence of In Vitro Grown Nili Ravi Buffalo Oocytes

In the present study, the response of Nili Ravi buffalo oocytes to recombinant human follicle stimulating hormone (rhFSH) (Organon) on meiotic maturation in vitro was examined. Oocytes were matured in vitro in medium containing either 0 or 0.05 IU/ ml rhFSH and the stage of nuclear maturation recorded after 24 hours. The percentage of oocytes in the control group undergoing germinal vesicle breakdown (GVBD) observed after 24 hours of culture was 29 % whereas as in rhFSH group the percentage was 10 % were at this stage (P< 0.001).Thus in the presence of rhFSH, a significantly greater number of oocytes had progressed to the more advanced stages of nuclear maturation. Indeed, the maturation of GV (Germinal Vesicle) stage oocytes to the metaphase II (M II) stage after 24 hours was significantly (P< 0.0001) increased by the addition of rhFSH (82 % VS 47 %). The percentage of degenerated oocytes after 24 hours of culture was 24 % in control group, whereas in rhFSH group the percentage was 8 % after 24 hours. Degeneration of the oocytes after 24 hours was not significantly (P = 0. 9361) decreased.

Shape Restoration of the Left Ventricle

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

Modification of Anodized Mg Alloy Surface By Pulse Condition for Biodegradable Material

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