Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 75

Search results for: In vivo

75 In vitro and in vivo Assessment of Cholinesterase Inhibitory Activity of the Bark Extracts of Pterocarpus santalinus L. for the Treatment of Alzheimer’s Disease

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

Abstract:

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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74 Hydrogel Based on Cellulose Acetate Used as Scaffold for Cell Growth

Authors: A. Maria G. Melero, A. M. Senna, J. A. Domingues, M. A. Hausen, E. Aparecida R. Duek, V. R. Botaro

Abstract:

A hydrogel from cellulose acetate cross linked with ethylenediaminetetraacetic dianhydride (HAC-EDTA) was synthesized by our research group, and submitted to characterization and biological tests. Cytocompatibility analysis was performed by confocal microscopy using human adipocyte derived stem cells (ASCs). The FTIR analysis showed characteristic bands of cellulose acetate and hydroxyl groups and the tensile tests evidence that HAC-EDTA present a Young’s modulus of 643.7 MPa. The confocal analysis revealed that there was cell growth at the surface of HAC-EDTA. After one day of culture the cells presented spherical morphology, which may be caused by stress of the sequestration of Ca2+ and Mg2+ ions at the cell medium by HAC-EDTA, as demonstrated by ICP-MS. However, after seven days and 14 days of culture, the cells present fibroblastoid morphology, phenotype expected by this cellular type. The results give efforts to indicate this new material as a potential biomaterial for tissue engineering, in the future in vivo approach.

Keywords: Cellulose acetate, hydrogel, biomaterial, cellular growth.

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73 X-Ray Fluorescence Molecular Imaging with Improved Sensitivity for Biomedical Applications

Authors: Guohua Cao, Xu Dong

Abstract:

X-ray Fluorescence Molecular Imaging (XFMI) holds great promise as a low-cost molecular imaging modality for biomedical applications with high chemical sensitivity. However, for in vivo biomedical applications, a key technical bottleneck is the relatively low chemical sensitivity of XFMI, especially at a reasonably low radiation dose. In laboratory x-ray source based XFMI, one of the main factors that limits the chemical sensitivity of XFMI is the scattered x-rays. We will present our latest findings on improving the chemical sensitivity of XFMI using excitation beam spectrum optimization. XFMI imaging experiments on two mouse-sized phantoms were conducted at three different excitation beam spectra. Our results show that the minimum detectable concentration (MDC) of iodine can be readily increased by five times via excitation spectrum optimization. Findings from this investigation could find use for in vivo pre-clinical small-animal XFMI in the future.

Keywords: Molecular imaging, X-ray fluorescence, chemical sensitivity, X-ray scattering.

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72 ELISA Based hTSH Assessment Using Two Sensitive and Specific Anti-hTSH Polyclonal Antibodies

Authors: Maysam Mard-Soltani, Mohamad Javad Rasaee, Saeed Khalili, Abdol Karim Sheikhi, Mehdi Hedayati

Abstract:

Production of specific antibody responses against hTSH is a cumbersome process due to the high identity between the hTSH and the other members of the glycoprotein hormone family (FSH, LH and HCG) and the high identity between the human hTSH and host animals for antibody production. Therefore, two polyclonal antibodies were purified against two recombinant proteins. Four possible ELISA tests were designed based on these antibodies. These ELISA tests were checked against hTSH and other glycoprotein hormones, and their sensitivity and specificity were assessed. Bioinformatics tools were used to analyze the immunological properties. After the immunogen region selection from hTSH protein, c terminal of B hTSH was selected and applied. Two recombinant genes, with these cut pieces (first: two repeats of C terminal of B hTSH, second: tetanous toxin+B hTSH C terminal), were designed and sub-cloned into the pET32a expression vector. Standard methods were used for protein expression, purification, and verification. Thereafter, immunizations of the white New Zealand rabbits were performed and the serums of them were used for antibody titration, purification and characterization. Then, four ELISA tests based on two antibodies were employed to assess the hTSH and other glycoprotein hormones. The results of these assessments were compared with standard amounts. The obtained results indicated that the desired antigens were successfully designed, sub-cloned, expressed, confirmed and used for in vivo immunization. The raised antibodies were capable of specific and sensitive hTSH detection, while the cross reactivity with the other members of the glycoprotein hormone family was minimum. Among the four designed tests, the test in which the antibody against first protein was used as capture antibody, and the antibody against second protein was used as detector antibody did not show any hook effect up to 50 miu/l. Both proteins have the ability to induce highly sensitive and specific antibody responses against the hTSH. One of the antibody combinations of these antibodies has the highest sensitivity and specificity in hTSH detection.

Keywords: hTSH, bioinformatics, protein expression, cross reactivity.

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71 Evaluation of Thrombolytic Activity of Zingiber cassumunar Roxb. and Thai Herbal Prasaplai Formula

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

Abstract:

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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70 Ultrasound Mechanical Index as a Parameter Affecting of the Ability of Proliferation of Cells

Authors: Z. Hormozi Moghaddam, M. Mokhtari-Dizaji, M. Movahedin, M. E. Ravari

Abstract:

Mechanical index (MI) is used for quantifying acoustic cavitation and the relationship between acoustic pressure and the frequency. In this study, modeling of the MI was applied to provide treatment protocol and to understand the effective physical processes on reproducibility of stem cells. The acoustic pressure and MI equations are modeled and solved to estimate optimal MI for 28, 40, 150 kHz and 1 MHz frequencies. Radial and axial acoustic pressure distribution was extracted. To validate the results of the modeling, the acoustic pressure in the water and near field depth was measured by a piston hydrophone. Results of modeling and experiments show that the model is consistent well to experimental results with 0.91 and 0.90 correlation of coefficient (p<0.05) for 1 MHz and 40 kHz. Low intensity ultrasound with 0.40 MI is more effective on the proliferation rate of the spermatogonial stem cells during the seven days of culture, in contrast, high MI has a harmful effect on the spermatogonial stem cells. This model provides proper treatment planning in vitro and in vivo by estimating the cavitation phenomenon.

Keywords: Ultrasound, mechanical index, modeling, stem cell.

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69 In vivo Therapeutic Potential of Biologically Synthesized Silver Nanoparticles

Authors: Kalakotla Shanker, G. Krishna Mohan

Abstract:

Nowadays, nanoparticles are being used in pharmacological studies for their exclusive properties such as small size, more surface area, biocompatibility and enhanced solubility. In view of this, the present study aimed to evaluate the antihyperglycemic potential of biologically synthesized silver nanoparticles (BSSNPs) and Gymnema sylvestre (GS) extract. The SEM and SEM analysis divulges that the BSSNPs were spherical in shape. EDAX spectrum exhibits peaks for the presence of silver, carbon, and oxygen atoms in the range of 1.0-3.1 keV. FT-IR reveals the binding properties of active bio-constituents responsible for capping and stabilizing BSSNPs. The results showed increased blood glucose, huge loss in body weight and downturn in plasma insulin. The GS extract (200 mg/kg, 400 mg/kg), BSSNPs (100 mg/kg, 200 mg/kg) and metformin 50 mg/kg were administered to the diabetic rats. BSSNPs at a dose level of 200 mg/kg (b.wt.p.o.) showed significant inhibition of (p<0.001) blood glucose levels as compared with GS extract treated group. The results obtained from study indicate that the BSSNP shows potent anti-diabetic activity.

Keywords: BSSNP, G.sylvetre, wistar rats, antihyperglycemic activity.

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68 Analysis of the Internal Mechanical Conditions in the Lower Limb Due to External Loads

Authors: Kent Salomonsson, Xuefang Zhao, Sara Kallin

Abstract:

Human soft tissue is loaded and deformed by any activity, an effect known as a stress-strain relationship, and is often described by a load and tissue elongation curve. Several advances have been made in the fields of biology and mechanics of soft human tissue. However, there is limited information available on in vivo tissue mechanical characteristics and behavior. Confident mechanical properties of human soft tissue cannot be extrapolated from e.g. animal testing. Thus, there is need for non invasive methods to analyze mechanical characteristics of soft human tissue. In the present study, the internal mechanical conditions of the lower limb, which is subject to an external load, is studied by use of the finite element method. A detailed finite element model of the lower limb is made possible by use of MRI scans. Skin, fat, bones, fascia and muscles are represented separately and the material properties for them are obtained from literature. Previous studies have been shown to address macroscopic deformation features, e.g. indentation depth, to a large extent. However, the detail in which the internal anatomical features have been modeled does not reveal the critical internal strains that may induce hypoxia and/or eventual tissue damage. The results of the present study reveals that lumped material models, i.e. averaging of the material properties for the different constituents, does not capture regions of critical strains in contrast to more detailed models.

Keywords: FEM, human soft tissue, indentation, properties.

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67 Formulation and Evaluation of Niosomes Containing an Antihypertensive Drug

Authors: Sunil Kamboj, Suman Bala, Vipin Saini

Abstract:

Niosomes were formulated with an aim of enhancing the oral bioavailability of losartan potassium and formulated in different molar ratios of surfactant, cholesterol and dicetyl phosphate. The formulated niosomes were found in range of 54.98 µm to 107.85 µm in size. Formulations with 1:1 ratio of surfactant and cholesterol have shown maximum entrapment efficiencies. Niosomes with sorbitan monostearate showed maximum drug release and zero order release kinetics, at the end of 24 hours. The in vivo study has shown the significant enhancement in oral bioavailability of losartan potassium in rats, after a dose of 10 mg/kg. The average relative bioavailability in relation with pure drug solution was found 2.56, indicates more than two fold increase in oral bioavailability. A significant increment in MRT reflects the release retarding ability of the vesicles. In conclusion, niosomes could be a promising delivery of losartan potassium with improved oral bioavailability and prolonged release profiles.

Keywords: Non-ionic surfactant vesicles, losartan potassium, oral bioavailability, controlled release.

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66 Phytochemical Screening, Antioxidant Activity and Lipid Profile Effects of Citrus reticulata Fruit Peel, Zingiber officinale Rhizome and Sesamum indicum Seed Extracts

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

Abstract:

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

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

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65 Antiangiogenic Potential of Phellodendron amurense Bark Extract Observed on Chorioallantoic Membrane

Authors: Ľudmila Ballová, Slavomír Kurhajec, Eva Petrovová, Jarmila Eftimová

Abstract:

Angiogenesis, a formation of new blood vessels from a pre-existing vasculature, plays an important role in pathologic processes such as the growth and metastasis of tumours. Tumours cannot grow beyond a few millimetres without blood supply from the newly formed blood vessels from the host tissue, a process called tumour-induced angiogenesis. The successful research of antiangiogenic treatment of cancer has focused on nutraceuticals with angiogenesis-modulating properties. Berberine, as a major active component of the bark of Phellodendron amurense Rupr., has shown antitumour activity by intervening into different steps of carcinogenesis. The influence of ethanolic extract of Phellodendron amurese bark on the angiogenesis was tested in vivo on chick chorioallantoic membrane (CAM). The irritancy of the CAM after the application of the crude bark extract dissolved in normal saline (10 mg/mL) was investigated on embryonic day 7. No significant signs of the irritancy, such as vasoconstriction, hyperaemia, haemorrhage or coagulation were observed which indicates the harmless character of the extract. A significant reduction in vessel sprouting and higher percentage of avascular zone was observed in the case of CAM treated with the extract in comparison with non-treated CAM (control), which is a proof of the antiangiogenic potential of the extract. These results could contribute to the development of novel drugs for the treatment of cancer or other diseases, in which angiogenesis plays a significant role.

Keywords: Angiogenesis, berberine, chorioallantoic membrane, Phellodendron amurense.

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64 The Toxicity of Doxorubicin with Nanotransporters

Authors: I. Blazkova, A. Moulick, V. Milosavljevic, P. Kopel, M. Vaculovicova, V. Adam, R. Kizek

Abstract:

Doxorubicin (DOX) is an anthracycline drug used to treat many cancer diseases. Similarly to other cytostatic drugs, DOX has serious side effects; the biggest obstacle is the cardiotoxicity. With the aim of lowering the negative side effects and to target the DOX into the tumor tissue, the different nanoparticles (NPs) are studied. The aim of this work was to synthetized different NPs and conjugated them with DOX and determine the binding capacity of the NPs. For this experiment, carbon nanotubes (CNTs), graphene oxide (GO), fullerene (FUL) and liposomes (LIP) were used. The highest binding capacity was observed in GO (85%). Subsequently the toxicity of NPs and NPs-DOX conjugates was analyzed in in vivo system (chicken embryos). Some NPs (GO) can increase the toxicity of DOX, whereas other NPs (LIP, CNTs) decrease DOX toxicity.

Keywords: Chicken embryos, Doxorubicin, Nanotransporters, Toxicity

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63 In vivo Alterations in Ruminal Parameters by Megasphaera elsdenii Inoculation on Subacute Ruminal Acidosis (SARA)

Authors: M. S. Alatas, H. D. Umucalilar

Abstract:

SARA is a common and serious metabolic disorder in early lactation in dairy cattle and in finishing beef cattle, caused by diets with high inclusion of cereal grain. This experiment was performed to determine the efficacy of Megasphaera elsdenii, a major lactate-utilizing bacterium in prevention/treatment of SARA in vivo. In vivo experimentation, it was used eight ruminally cannulated rams and it was applied the rapid adaptation with the mixture of grain based on wheat (80% wheat, 20% barley) and barley (80% barley, 20% wheat). During the systematic adaptation, it was followed the probability of SARA formation by being measured the rumen pH with two hours intervals after and before feeding. After being evaluated the data, it was determined the ruminal pH ranged from 5.2-5.6 on the condition of feeding with 60 percentage of grain mixture based on barley and wheat, that assured the definite form of subacute acidosis. In four days SARA period, M. elsdenii (1010 cfu ml-1) was inoculated during the first two days. During the SARA period, it was observed the decrease of feed intake with M. elsdenii inoculation. Inoculation of M. elsdenii was caused to differentiation of rumen pH (P<0.0001), while it was found the pH level approximately 5.55 in animals applied the inoculation, it was 5.63 pH in other animals. It was observed that total VFA with the bacterium inoculation tended to change in terms of grain feed (P<0.07). It increased with the effect of total VFA inoculation in barley based diet, but it was more stabilized in wheat based diet. Bacterium inoculation increased the ratio of propionic acid (18.33%-21.38%) but it caused to decrease the butyric acid, and acetic/propionic acid. During the rapid adaptation, the concentration of lactic acid in the rumen liquid increased depending upon grain level (P<0.0001). On the other hand bacterium inoculation did not have an effect on concentration of lactic acid. M. elsdenii inoculation did not affect ruminal ammonia concentration. In the group that did not apply inoculation, the level of ruminal ammonia concentration was higher than the others applied inoculation. M. elsdenii inoculation did not changed protozoa count in barley-based diet whereas it decreased in wheat-based diet. When it is generally evaluated, it is seen that M. elsdenii inoculation has not a positive impact on rumen parameters. Therefore, to reveal the full impact of the inoculation with different strains, feedstuffs and animal groups, further research is required.

Keywords: In vivo, subactute ruminal acidosis, Megasphaera elsdenii, rumen fermentation.

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62 A New Method to Enhance Contrast of Electron Micrograph of Rat Tissues Sections

Authors: Lise P. Labéjof, Raiza S. P. Bizerra, Galileu B. Costa, Thaísa B. dos Santos

Abstract:

This report presents an alternative technique of application of contrast agent in vivo, i.e. before sampling. By this new method the electron micrograph of tissue sections have an acceptable contrast compared to other methods and present no artifact of precipitation on sections. Another advantage is that a small amount of contrast is needed to get a good result given that most of them are expensive and extremely toxic.

Keywords: Image quality, Microscopy research, Staining technique, Ultrathin section.

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61 Influence of Cyperus rotundus Active Principles Inhibit Viral Multiplication and Stimulate Immune System in Indian White Shrimp Fenneropenaeus indicus against White Spot Syndrome Virus Infection

Authors: T. Citarasu, M. Michaelbabu V. N. Vakharia

Abstract:

The rhizome of Java grass, Cyperus rotundus was extracted different organic polar and non-polar solvents and performed the in vitro antiviral and immunostimulant activities against White Spot Syndrome Virus (WSSV) and Vibrio harveyi respectively. Based on the initial screening the ethyl acetate extract of C. rotundus was strong activities and further it was purified through silica column chromatography and the fractions were screened again for antiviral and immunostimulant activity. Among the different fractions screened against the WSSV and V. harveyi, the fractions, FIII to FV had strong activities. In order to study the in vivo influence of C. rotundus, the fractions (F-III to FV) were pooled and delivered to the F. indicus through artificial feed for 30 days. After the feeding trail the experimental and control diet fed F. indicus were challenged with virulent WSSV and studied the survival, molecular diagnosis, biochemical, haematological, and immunological parameters. Surprisingly, the pooled fractions (F-IV to FVI) incorporated diets helped to significantly (P<0.01) suppressed viral multiplication, showed significant (P<0.01) differences in protein and glucose levels, improved total haemocyte count (THC), coagulase activity, significantly increased (P <= 0.001) prophenol oxidase and intracellular superoxide anion production compared to the control shrimps. Based on the results, C. rotundus extracts effectively suppressed WSSV multiplication and improve the immune system in F. indicus against WSSV infection and this knowledge will helps to develop novel drugs from C. rotundus against WSSV.

Keywords: Antiviral drugs, Cyperus rotundus, Fenneropenaeus indicus, WSSV.

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60 Probiotic Potential and Antimicrobial Activity of Enterococcus faecium Isolated from Chicken Caecal and Fecal Samples

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

Abstract:

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

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

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59 A Piscan Ulcerative Aeromonas Infection

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

Abstract:

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

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

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58 In vivo Iron Availability and Profile Lipid Composition in Anemic Rats Fed on Diets with Black Rice Bran Extract

Authors: E. P. Nurlaili, M. Astuti, Y. Marsono, S. Naruki

Abstract:

Iron is an essential nutrient with limited bioavailability. Nutritional anemia caused mainly by iron deficiency is the most recognized nutritional problem in both countries as well as affluent societies. Rice (Oryza sativa L.) has become the most important cereal crop for the improvement of human health due to the starch, protein, oil, and the majority of micronutrients, particularly in Asian countries. In this study, the iron availability and profile lipid were evaluated for the extracts from Cibeusi varieties (black rices) of ancient rice brans. Results: The quality of K, B, R, E diets groups shows the same effect on the growth of rats. Hematocrit and MCHC levels of rats fed K, B, R and E diets were not significantly (P<0.05). MCV and MCH levels of rats K, B, R were significantly (P<0.05) with E groups but rats K, B, R were not significantly (P<0.05). The iron content in the serum of rats fed with K, B, R and E diets were not significantly (P<0.05). The highest level of iron in the serum was founded in the B group. The iron content in the liver of rats fed with K, B, R and E diets were not significantly (P<0.05). The highest level of iron in the liver was founded in the R group. HDL cholesterol levels were significantly (P<0.05) between rats of fed B, E with K, R, but K and R were not significantly (P<0.05). LDL cholesterol levels of rats fed K and E significantly (P<0.05) with B and R. Conclusions: the bran of pigmented rice varieties has, with some exceptions, greater antioxidant and free-radical scavenging activities. The results also show that pigmented rice extracts acted as prooxidants in the lipid peroxidation assay, possibly by mechanisms described for the pro-oxidant activities of tocopherol and ascorbic. Pigmented rice bran extracts more effectively increases iron stores and reduces the prevalence of iron deficiency.

Keywords: Anemia, black rice bran extract, iron, profile lipid.

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57 Antioxidative Potential of Aqueous Extract of Ocimum americanum L. Leaves: An in vitro and in vivo Evaluation

Authors: B. T. Aluko, O. I. Oloyede

Abstract:

Ocimum americanum L (Lamiaceae) is an annual herb that is native to tropical Africa. The in vitro and in vivo antioxidant activity of its aqueous extract was carefully investigated by assessing the DPPH radical scavenging activity, ABTS radical scavenging activity and hydrogen peroxide radical scavenging activity. The reducing power, total phenol, total flavonoids and flavonols content of the extract were also evaluated. The data obtained revealed that the extract is rich in polyphenolic compounds and scavenged the radicals in a concentration dependent manner. This was done in comparison with the standard antioxidants such as BHT and Vitamin C. Also, the induction of oxidative damage with paracetamol (2000 mg/kg) resulted in the elevation of lipid peroxides and significant (P < 0.05) decrease in activities of superoxide dismutase, glutathione peroxidase, glutathione reductase and catalase in the liver and kidney of rats. However, the pretreatment of rats with aqueous extract of O. americanum leaves (200 and 400 mg/kg) and silymarin (100 mg/kg) caused a significant (P < 0.05) reduction in the values of lipid peroxides and restored the levels of antioxidant parameters in these organs. These findings suggest that the leaves of O. americanum have potent antioxidant properties which may be responsible for its acclaimed folkloric uses.

Keywords: Antioxidants, free radicals, Ocimum americanum, scavenging activity.

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56 Evaluation of the Laser and Partial Vibration Stimulation on Osteoporosis

Authors: Ji Hyung Park, Dong-Hyun Seo, Young-Jin Jung, Han Sung Kim

Abstract:

The aim of this study is to evaluate the effects of the laser and partial vibration stimulation on the mice tibia with morphological characteristics. Twenty female C57BL/6 mice (12 weeks old) were used for the experiment. The study was carried out on four groups of animals each consisting of five mice. Four groups of mice were ovariectomized. Animals were scanned at 0 and 2 weeks after ovariectomy by using micro computed tomography to estimate morphological characteristics of tibial trabecular bone. Morphological analysis showed that structural parameters of multi-stimuli group appear significantly better phase in BV/TV, BS/BV, Tb.Th, Tb.N, Tb.Sp, and Tb.pf than single stimulation groups. However, single stimulation groups didn’t show significant effect on tibia with Sham group. This study suggests that multi-stimuli may restrain the change as the degenerate phase on osteoporosis in the mice tibia.

Keywords: Laser, Partial Vibration, Osteoporosis, in vivo micro-CT, mice.

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55 In vivo Antidiabetic and Antioxidant Potential of Pseudovaria macrophylla Extract

Authors: Aditya Arya, Hairin Taha, Ataul Karim Khan, Nayiar Shahid, Hapipah Mohd Ali, Mustafa Ali Mohd

Abstract:

This study has investigated the antidiabetic and antioxidant potential of Pseudovaria macrophylla bark extract on streptozotocin–nicotinamide induced type 2 diabetic rats. LCMSQTOF and NMR experiments were done to determine the chemical composition in the methanolic bark extract. For in vivo experiments, the STZ (60 mg/kg/b.w, 15 min after 120 mg/kg/1 nicotinamide, i.p.) induced diabetic rats were treated with methanolic extract of Pseuduvaria macrophylla (200 and 400 mg/kg·bw) and glibenclamide (2.5 mg/kg) as positive control respectively. Biochemical parameters were assayed in the blood samples of all groups of rats. The pro-inflammatory cytokines, antioxidant status and plasma transforming growth factor βeta-1 (TGF-β1) were evaluated. The histological study of the pancreas was examined and its expression level of insulin was observed by immunohistochemistry. In addition, the expression of glucose transporters (GLUT 1, 2 and 4) were assessed in pancreas tissue by western blot analysis. The outcomes of the study displayed that the bark methanol extract of Pseuduvaria macrophylla has potentially normalized the elevated blood glucose levels and improved serum insulin and C-peptide levels with significant increase in the antioxidant enzyme, reduced glutathione (GSH) and decrease in the level of lipid peroxidation (LPO). Additionally, the extract has markedly decreased the levels of serum pro-inflammatory cytokines and transforming growth factor beta-1 (TGF-β1). Histopathology analysis demonstrated that Pseuduvaria macrophylla has the potential to protect the pancreas of diabetic rats against peroxidation damage by downregulating oxidative stress and elevated hyperglycaemia. Furthermore, the expression of insulin protein, GLUT-1, GLUT-2 and GLUT-4 in pancreatic cells was enhanced. The findings of this study support the anti-diabetic claims of Pseudovaria macrophylla bark.

Keywords: Diabetes mellitus, Pseuduvaria macrophylla, alkaloids, caffeic acid.

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54 Lactic Acid-Chitosan Films’ Properties and Their in vivo Wound Healing Activity

Authors: T. S. Moe, T. A. Khaing

Abstract:

Chitosan is a derivative of chitin, a compound usually isolated from the shells of some crustaceans such as crab, lobster and shrimp. It has biocompatible, biodegradable, and antimicrobial properties. To use these properties of chitosan in biomedical fields, chitosan films (1%, 2%, 3% and 4%) were prepared by using l% lactic acid as solvent. The effects of chitosan films on tensile strength, elongation at break, degree of swelling, thickness, morphology, allergic and irritation reactions and antibacterial property were evaluated. Staphylococcus aureus and Escherichia coli were used as tested microorganisms. In vivo wound healing activities of chitosan films were investigated using mice model. As results, Chitosan films have similar appearance and good swelling properties and 4% chitosan film showed the better swelling activity and the greatest elongation ratio than the other chitosan films. They also showed their good activity of wound healing in mice model. Moreover, the results showed that the films did not produce any unwilling symptoms (allergy or irritation). In conclusion, it is evident that the chitosan film has the potentiality to use as wound healing biofilms in the biomedical fields.

Keywords: Chitosan, wound healing, antibacterial activity.

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53 Preparation and in vivo Assessment of Nystatin-Loaded Solid Lipid Nanoparticles for Topical Delivery against Cutaneous Candidiasis

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

Abstract:

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

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

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52 A Numerical Model for Simulation of Blood Flow in Vascular Networks

Authors: Houman Tamaddon, Mehrdad Behnia, Masud Behnia

Abstract:

An accurate study of blood flow is associated with an accurate vascular pattern and geometrical properties of the organ of interest. Due to the complexity of vascular networks and poor accessibility in vivo, it is challenging to reconstruct the entire vasculature of any organ experimentally. The objective of this study is to introduce an innovative approach for the reconstruction of a full vascular tree from available morphometric data. Our method consists of implementing morphometric data on those parts of the vascular tree that are smaller than the resolution of medical imaging methods. This technique reconstructs the entire arterial tree down to the capillaries. Vessels greater than 2 mm are obtained from direct volume and surface analysis using contrast enhanced computed tomography (CT). Vessels smaller than 2mm are reconstructed from available morphometric and distensibility data and rearranged by applying Murray’s Laws. Implementation of morphometric data to reconstruct the branching pattern and applying Murray’s Laws to every vessel bifurcation simultaneously, lead to an accurate vascular tree reconstruction. The reconstruction algorithm generates full arterial tree topography down to the first capillary bifurcation. Geometry of each order of the vascular tree is generated separately to minimize the construction and simulation time. The node-to-node connectivity along with the diameter and length of every vessel segment is established and order numbers, according to the diameter-defined Strahler system, are assigned. During the simulation, we used the averaged flow rate for each order to predict the pressure drop and once the pressure drop is predicted, the flow rate is corrected to match the computed pressure drop for each vessel. The final results for 3 cardiac cycles is presented and compared to the clinical data.

Keywords: Blood flow, Morphometric data, Vascular tree, Strahler ordering system.

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51 Systematic Identification and Quantification of Substrate Specificity Determinants in Human Protein Kinases

Authors: Manuel A. Alonso-Tarajano, Roberto Mosca, Patrick Aloy

Abstract:

Protein kinases participate in a myriad of cellular processes of major biomedical interest. The in vivo substrate specificity of these enzymes is a process determined by several factors, and despite several years of research on the topic, is still far from being totally understood. In the present work, we have quantified the contributions to the kinase substrate specificity of i) the phosphorylation sites and their surrounding residues in the sequence and of ii) the association of kinases to adaptor or scaffold proteins. We have used position-specific scoring matrices (PSSMs), to represent the stretches of sequences phosphorylated by 93 families of kinases. We have found negative correlations between the number of sequences from which a PSSM is generated and the statistical significance and the performance of that PSSM. Using a subset of 22 statistically significant PSSMs, we have identified specificity determinant residues (SDRs) for 86% of the corresponding kinase families. Our results suggest that different SDRs can function as positive or negative elements of substrate recognition by the different families of kinases. Additionally, we have found that human proteins with known function as adaptors or scaffolds (kAS) tend to interact with a significantly large fraction of the substrates of the kinases to which they associate. Based on this characteristic we have identified a set of 279 potential adaptors/scaffolds (pAS) for human kinases, which is enriched in Pfam domains and functional terms tightly related to the proposed function. Moreover, our results show that for 74.6% of the kinase–pAS association found, the pAS colocalize with the substrates of the kinases they are associated to. Finally, we have found evidence suggesting that the association of kinases to adaptors and scaffolds, may contribute significantly to diminish the in vivo substrate crossed-specificity of protein kinases. In general, our results indicate the relevance of several SDRs for both the positive and negative selection of phosphorylation sites by kinase families and also suggest that the association of kinases to pAS proteins may be an important factor for the localization of the enzymes with their set of substrates.

Keywords: Kinase, phosphorylation, substrate specificity, adaptors, scaffolds, cellular colocalization.

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50 The Efficacy of Andrographis paniculata and Chromolaena odorata Plant Extract against Malaria Parasite

Authors: Funmilola O. Omoya, Abdul O. Momoh

Abstract:

Malaria constitutes one of the major health problems in Nigeria. One of the reasons attributed for the upsurge was the development of resistance of Plasmodium falciparum and the emergence of multi-resistant strains of the parasite to anti-malaria drugs. A continued search for other effective, safe and cheap plantbased anti-malaria agents thus becomes imperative in the face of these difficulties. The objective of this study is therefore to evaluate the in vivo anti-malarial efficacy of ethanolic extracts of Chromolaena odorata and Androgaphis paniculata leaves. The two plants were evaluated for their anti-malaria efficacy in vivo in a 4-day curative test assay against Plasmodium berghei strain in mice. The group treated with 500mg/ml dose of ethanolic extract of A. paniculata plant showed parasite suppression with increase in Packed Cell Volume (PCV) value except day 3 which showed a slight decrease in PCV value. During the 4-day curative test, an increase in the PCV values, weight measurement and zero count of Plasmodium berghei parasite values was recorded after day 3 of drug administration. These results obtained in group treated with A. paniculata extract showed anti-malarial efficacy with higher mortality rate in parasitaemia count when compared with Chromolaena odorata group. These results justify the use of ethanolic extracts of A. paniculata plant as medicinal herb used in folklore medicine in the treatment of malaria.

Keywords: Anti-malaria, Curative, Plant-based anti-malaria agents.

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49 The Antibacterial Efficacy of Gold Nanoparticles Derived from Gomphrena celosioides and Prunus amygdalus (Almond) Leaves on Selected Bacterial Pathogens

Authors: M. E. Abalaka, S. Y. Daniyan, S. O. Adeyemo, D. Damisa

Abstract:

Gold nanoparticles (AuNPs) have gained increasing interest in recent times. This is greatly due to their special features, which include unusual optical and electronic properties, high stability and biological compatibility, controllable morphology and size dispersion, and easy surface functionalization. In typical synthesis, AuNPs were produced by reduction of gold salt AuCl4 in an appropriate solvent. A stabilizing agent was added to prevent the particles from aggregating. The antibacterial activity of different sizes of gold nanoparticles was investigated against Staphylococcus aureus, Salmonella typhi and Pseudomonas pneumonia using the disk diffusion method in a Müeller–Hinton Agar. The Au-NPs were effective against all bacteria tested. That the Au-NPs were successfully synthesized in suspension and were used to study the antibacterial activity of the two medicinal plants against some bacterial pathogens suggests that Au-NPs can be employed as an effective bacteria inhibitor and may be an effective tool in medical field. The study clearly showed that the Au-NPs exhibiting inhibition towards the tested pathogenic bacteria in vitro could have the same effects in vivo and thus may be useful in the medical field if well researched into.

Keywords: Gold Nanoparticles, Gomphrena celesioides, Prunus amygdalus, Pathogens.

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48 In vitro and in vivo Anticholinesterase Activity of the Volatile Oil of the Aerial Parts of Ocimum basilicum L. and O. africanum Lour. Growing in Egypt

Authors: M. G. Tadros, S. M. Ezzat, M. M. Salama, M. A. Farag

Abstract:

In this study, the in vitro anticholinesterase activity of
the volatile oils of both O. basilicum and O. africanum was
investigated and both samples showed significant activity. The major
constituents of the two oils were isolated using several column
chromatographies. Linalool, 1,8-cineol and eugenol were isolated
from the volatile oil of O. basilicum and camphor was isolated from
the volatile oil of O. africanum. The anticholinesterase activities of
the isolated compounds were also evaluated where 1,8-cineol showed
the highest inhibitory activity followed by camphor. To confirm these
activities, learning and memory enhancing effects were tested in
mice. Memory impairment was induced by scopolamine, a
cholinergic muscarinic receptor antagonist. Anti-amnesic effects of
both volatile oils and their terpenoids were investigated by the
passive avoidance task in mice. We also examined their effects on
brain acetylcholinesterase activity. Results showed that scopolamineinduced
cognitive dysfunction was significantly attenuated by
administration of the volatile oils and their terpenoids, eugenol and
camphor, in the passive avoidance task and inhibited brain
acetylcholinesterase activity. These results suggest that O. basilicum
and O. africanum volatile oils can be good candidates for further
studies on Alzheimer’s disease via their acetylcholinesterase
inhibitory actions.

Keywords: Acetylcholinesterase, Ocimum africanum, Ocimum basilicum, passive avoidance.

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47 Study of Functional Relevant Conformational Mobility of β-2 Adrenoreceptor by Means of Molecular Dynamics Simulation

Authors: G. V. Novikov, V. S. Sivozhelezov, S. S. Kolesnikov, K. V. Shaitan

Abstract:

The study reports about the influence of binding of orthosteric ligands as well as point mutations on the conformational dynamics of β-2-adrenoreceptor. Using molecular dynamics simulation we found that there was a little fraction of active states of the receptor in its apo (ligand free) ensemble corresponded to its constitutive activity. Analysis of MD trajectories indicated that such spontaneous activation of the receptor is accompanied by the motion in intracellular part of its alpha-helices. Thus receptor’s constitutive activity directly results from its conformational dynamics. On the other hand the binding of a full agonist resulted in a significant shift of the initial equilibrium towards its active state. Finally, the binding of the inverse agonist stabilized the receptor in its inactive state. It is likely that the binding of inverse agonists might be a universal way of constitutive activity inhibition in vivo. Our results indicate that ligand binding redistribute pre-existing conformational degrees of freedom (in accordance to the Monod-Wyman-Changeux-Model) of the receptor rather than cause induced fit in it. Therefore, the ensemble of biologically relevant receptor conformations is encoded in its spatial structure, and individual conformations from that ensemble might be used by the cell in conformity with the physiological behavior.

Keywords: Seven-transmembrane receptors, constitutive activity, activation, x-ray crystallography, principal component analysis, molecular dynamics simulation.

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46 In vitro and in vivo Anticancer Activity of Nanosize Zinc Oxide Composites of Doxorubicin

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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