Tuberculin, Tetanus Immunoglobulin and DPT Vaccine as an Avian in vivo T- Lymphocyte Mitogens

The avian phytohaemagglutinin skin test is being proved as an in vivo system for the evaluation an avian in vivo T cell mitogenicity. The test system was one week old Gallus domesticus broiler Chickens. Five replicates were done for each of the whole, 1:10 dilutions of each of 0.05 IU tuberculin, tetanus immunoglobulin and DPT vaccine as test materials. The evaluation parameters were the skin indurations and lymphoblast percentages in bone marrow lymphocytes. Tuberculin indurations were 2.06 and 1.26mm for 0.05 IU respectively while lymphoblast percent were 0.234 and 0.1 accordingly. The skin indurations of 135mg/ml and 1.35mg/ml tetanus immunoglobulin were 4.86 and 3.96mm while lymphoblast percentages were 0.3 and 0.14 respectively. The whole DPT and 1:10 concentration were with 4.5 and 3.2mm while their lymphoblast percentages were 0.28 and 0.12 accordingly. Thus the mitogenicity of the test materials was of dependant type.

Influence of Calcium Intake Level to Osteoporptic Vertebral bone and Degenerated Disc in Biomechanical Study

The aim of the present study is to analyze the generation of osteoporotic vertebral bone induced by lack of calcium during growth period and analyze its effects for disc degeneration, based on biomechanical and histomorphometrical study. Mechanical and histomorphological characteristics of lumbar vertebral bones and discs of rats with calcium free diet (CFD) were detected and tracked by using high resolution in-vivo micro-computed tomography (in-vivo micro-CT), finite element (FE) and histological analysis. Twenty female Sprague-Dawley rats (6 weeks old, approximate weight 170g) were randomly divided into two groups (CFD group: 10, NOR group: 10). The CFD group was maintained on a refmed calcium-controlled semisynthetic diet without added calcium, to induce osteoporosis. All lumbar (L 1-L6) were scanned by using in vivo micro-CT with 35i.un resolution at 0, 4, 8 weeks to track the effects of CFD on the generation of osteoporosis. The fmdings of the present study indicated that calcium insufficiency was the main factor in the generation of osteoporosis and it induced lumbar vertebral disc degeneration. This study is a valuable experiment to firstly evaluate osteoporotic vertebral bone and disc degeneration induced by lack of calcium during growth period from a biomechanical and histomorphometrical point of view.

A Comparative Study of Transient Flow through Cerebral Aneurysms using CFD

The recent advances in computational fluid dynamics (CFD) can be useful in observing the detailed hemodynamics in cerebral aneurysms for understanding not only their formation and rupture but also for clinical evaluation and treatment. However, important hemodynamic quantities are difficult to measure in vivo. In the present study, an approximate model of normal middle cerebral artery (MCA) along with two cases consisting broad and narrow saccular aneurysms are analyzed. The models are generated in ANSYS WORKBENCH and transient analysis is performed in ANSYS-CFX. The results obtained are compared for three cases and agree well with the available literature.

Characterization of the O.ul-mS952 Intron:A Potential Molecular Marker to Distinguish Between Ophiostoma Ulmi and Ophiostoma Novo-Ulmi Subsp. Americana

The full length mitochondrial small subunit ribosomal (mt-rns) gene has been characterized for Ophiostoma novo-ulmi subspecies americana. The gene was also characterized for Ophiostoma ulmi and a group II intron was noted in the mt-rns gene of O. ulmi. The insertion in the mt-rns gene is at position S952 and it is a group IIB1 intron that encodes a double motif LAGLIDADG homing endonuclease from an open reading frame located within a loop of domain III. Secondary structure models for the mt-rns RNA of O. novo-ulmi subsp. americana and O. ulmi were generated to place the intron within the context of the ribosomal RNA. The in vivo splicing of the O.ul-mS952 group II intron was confirmed with reverse transcription-PCR. A survey of 182 strains of Dutch Elm Diseases causing agents showed that the mS952 intron was absent in what is considered to be the more aggressive species O. novo-ulmi but present in strains of the less aggressive O. ulmi. This observation suggests that the O.ul-mS952 intron can be used as a PCR-based molecular marker to discriminate between O. ulmi and O. novo-ulmi subsp. americana.

In vivo Histomorphometric and Corrosion Analysis of Ti-Ni-Cr Shape Memory Alloys in Rabbits

A series of Ti based shape memory alloys with composition of Ti50Ni49Cr1, Ti50Ni47Cr3 and Ti50Ni45Cr5 were developed by vacuum arc-melting under a purified argon atmosphere. The histometric and corrosion evaluation of Ti-Ni-Cr shape memory alloys have been considered in this research work. The alloys were developed by vacuum arc melting and implanted subcutaneously in rabbits for 4, 8 and 12 weeks. Metallic implants were embedded in order to determine the outcome of implantation on histometric and corrosion evaluation of Ti-Ni-Cr metallic strips. Encapsulating membrane formation around the alloys was minimal in the case of all materials. After histomorphometric analyses it was possible to demonstrate that there were no statistically significant differences between the materials. Corrosion rate was also determined in this study which is within acceptable range. The results showed the Ti- Ni-Cr alloy was neither cytotoxic, nor have any systemic reaction on living system in any of the test performed. Implantation shows good compatibility and a potential of being used directly in vivo system.

The Role of Immunogenic Adhesin Vibrio alginolyticus 49 k Da to Molecule Expression of Major Histocompatibility Complex on Receptors of Humpback Grouper Cromileptes altivelis

The purpose of research was to know the role of immunogenic protein of 49 kDa from V.alginolyticus which capable to initiate molecule expression of MHC Class II in receptor of Cromileptes altivelis. The method used was in vivo experimental research through testing of immunogenic protein 49 kDa from V.alginolyticus at Cromileptes altivelis (size of 250 - 300 grams) using 3 times booster by injecting an immunogenic protein in a intramuscular manner. Response of expressed MHC molecule was shown using immunocytochemistry method and SEM. Results indicated that adhesin V.alginolyticus 49 kDa which have immunogenic character could trigger expression of MHC class II on receptor of grouper and has been proven by staining using immunocytochemistry and SEM with labeling using antibody anti MHC (anti mouse). This visible expression based on binding between epitopes antigen and antibody anti MHC in the receptor. Using immunocytochemistry, intracellular response of MHC to in vivo induction of immunogenic adhesin from V.alginolyticus was shown.

Why We Are Taller in the Morning than Going to Bed at Night – An in vivo and in vitro Study

Intradiscal and intervertebral pressure transducers were developed. They were used to map the pressures in the nucleus and within the annulus of the human spinal segments. Their stressrelaxation were recorded over a period of time for nucleus pressure, applied load, and peripherial strain against time. The results show that for normal discs, pressures in the nucleus are viscoelastic in nature with the applied compressive load. Mechanical strains which develop around the periphery of the vertebral body are also viscoelastic with the applied compressive load. Applied compressive load against time also shows viscoelastic behavior. However, annulus does not respond viscoelastically with the applied load. It showed a linear response to compressive loading.

Analysis of DNA-Recognizing Enzyme Interaction using Deaminated Lesions

Deaminated lesions were produced via nitrosative oxidation of natural nucleobases; uracul (Ura, U) from cytosine (Cyt, C), hypoxanthine (Hyp, H) from adenine (Ade, A), and xanthine (Xan, X) and oxanine (Oxa, O) from guanine (Gua, G). Such damaged nucleobases may induce mutagenic problems, so that much attentions and efforts have been poured on the revealing of their mechanisms in vivo or in vitro. In this study, we employed these deaminated lesions as useful probes for analysis of DNA-binding/recognizing proteins or enzymes. Since the pyrimidine lesions such as Hyp, Oxa and Xan are employed as analogues of guanine, their comparative uses are informative for analyzing the role of Gua in DNA sequence in DNA-protein interaction. Several DNA oligomers containing such Hyp, Oxa or Xan substituted for Gua were designed to reveal the molecular interaction between DNA and protein. From this approach, we have got useful information to understand the molecular mechanisms of the DNA-recognizing enzymes, which have not ever been observed using conventional DNA oligomer composed of just natural nucleobases.

Endothelial Specificity of ICAM2, Flt-1, and Tie2 Promoters In Vitro and In Vivo

To identify an endothelial cell-specific promoter suitable for vascular-specific targeting, we tested five promoters in vitro--Tie2SE, Tie2LE, ICAM2, Flt-1 and vWF--for promoter activity and specificity in endothelial cells, smooth muscle cells and non-vascular resident cells as well as tissues. These promoters, except for vWF, exhibited good endothelial activity and specificity in vitro. In a syngenic heart transplantation model, the ICAM2 promoter was variably functional in coronary endothelial cells of donor hearts. Thus, the ICAM2, Flt-1, Tie2SE and Tie2LE promoters hold promise for endothelial-specific targeting, but in vitro expression may not predict in vivo expression.

Optical Coherence Tomography Combined with the Confocal Microscopy Method and Fluorescence for Class V Cavities Investigations

The purpose of this study is to present a non invasive method for the marginal adaptation evaluation in class V composite restorations. Standardized class V cavities, prepared in human extracted teeth, were filled with Premise (Kerr) composite. The specimens were thermo cycled. The interfaces were examined by Optical Coherence Tomography method (OCT) combined with the confocal microscopy and fluorescence. The optical configuration uses two single mode directional couplers with a superluminiscent diode as the source at 1300 nm. The scanning procedure is similar to that used in any confocal microscope, where the fast scanning is enface (line rate) and the depth scanning is much slower (at the frame rate). Gaps at the interfaces as well as inside the composite resin materials were identified. OCT has numerous advantages which justify its use in vivo as well as in vitro in comparison with conventional techniques.

Effects of a Methanol Fraction of the Leaves of Leonotis leonurus on the Blood Pressure and Heart Rate of Normotensive Male Wistar Rats

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

Osteogenesis by Dextran Coating on and among Fibers of a Polyvinyl Formal Sponge

A scaffold is necessary for tooth regeneration because of its three-dimensional geometry. For restoration of defect, it is necessary for the scaffold to be prepared in the shape of the defect. Sponges made from polyvinyl alcohol with formalin cross-linking (PVF sponge) have been used for scaffolds for bone formation in vivo. To induce osteogenesis within the sponge, methods of growing rat bone marrow cells (rBMCs) among the fiber structures in the sponge might be considered. Storage of rBMCs among the fibers in the sponge coated with dextran (10 kDa) was tried. After seeding of rBMCs to PVF sponge immersed in dextran solution at 2 g/dl concentration, osteogenesis was recognized in subcutaneously implanted PVF sponge as a scaffold in vivo. The level of osteocalcin was 25.28±5.71 ng/scaffold and that of Ca was 129.20±19.69 µg/scaffold. These values were significantly higher than those in sponges without dextran coating (p

Realignment of f-actin Cytoskeleton in Osteocytes after Mechanical Loading

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

Anticoagulatory Role of an Ergot Mesylate: Hydergine

Thrombosis can be life threatening, necessitating therefore its instant treatment. Hydergine, a nootropic agent is used as a cognition enhancer in stroke patients but relatively little is known about its anti-thrombolytic effect. To investigate this aspect, in vivo and ex vivo experiments were designed and conducted. Three groups of rats were injected 1.5mg, 3.0mg and 4.5mg hydergine intraperitonealy with and without prior exposure to fresh plasma. Positive and negative controls were run in parallel. Animals were sacrificed after 1.5hrs and BT, CT, PT, INR, APTT, plasma calcium levels were estimated. For ex vivo analyses, each 1ml blood aspirated was exposed to 0.1mg, 0.2mg, 0.3mg dose of hydergine with parallel controls. Parameters analyzed were as above. Statistical analysis was through one-way ANOVA. Dunken-s and Tukey-s tests provided intra-group variance. BT, CT, PT, INR and APTT increased while calcium levels dropped significantly (P

Nanobiocomposites with Enhanced Cell Proliferation and Improved Mechanical Properties Based on Organomodified-Nanoclay and Silicone Rubber

Bionanotechnology deals with nanoscopic interactions between nanostructured materials and biological systems. Polymer nanocomposites with optimized biological activity have attracted great attention. Nanoclay is considered as reinforcing nanofiller in manufacturing of high performance nanocomposites. In current study, organomodified-nanoclay with negatively charged silicate layers was incorporated into biomedical grade silicone rubber. Nanoparticle loading has been tailored to enhance cell behavior. Addition of nanoparticles led to improved mechanical properties of substrate with enhanced strength and stiffness while no toxic effects was observed. Results indicated improved viability and proliferation of cells by addition of nanofillers. The improved mechanical properties of the matrix result in proper cell response through adjustment and arrangement of cytoskeletal fibers. Results can be applied in tissue engineering when enhanced substrates are required for improvement of cell behavior for in vivo applications.

Polyphenolic Profile and Antioxidant Activities of Nigella Sativa Seed Extracts In Vitro and In Vivo

Nigella sativa L. is an aromatic plant belonging to the family Ranunculaceae. It has been used traditionally, especially in the middle East and India, for the treatment of asthma, cough, bronchitis, headache, rheumatism, fever, influenza and eczema. Several biological activities have been reported in Nigella sativa seeds, including antioxidant. In this context we tried to estimate the antioxidant activity of various extracts prepared from Nigella sativa seeds, methanolic extract (ME), chloroformic extract (CE), hexanic extract (HE : fixed oil), ethyl acetate extract (EAE) water extract (WE). The Folin-Ciocalteu assay showed that CE and EAE contained high level of phenolic compounds 81.31 and 72.43μg GAE/mg of extract respectively. Similarly, the CE and EAE exhibited the highest DPPH radical scavenging activity, with IC50 values of 106.56μg/ml and 121.62μg/ml respectively. In addition, CE and HE showed the most scavenging activity against superoxide radical generated in the PMS-NADH-NBT system with respective IC50 values of 361.86 μg/ml and 371.80 μg/ml, which is comparable to the activity of the standard antioxidant BHT (344.59 μg/ml). Ferrous ion chelating capacity assay showed that WE, EAE and ME are the most active with 40.57, 39.70 and 22.02 mg EDTA-E/g of extract. The inhibition of linoleic acid/ß-carotene coupled oxidation was estimated by ßcarotene bleaching assay, this showed a highest relative antioxidant activity with CE and EAE (69.82% of inhibition). The antioxidant activities of the methanolic extract and the fixed oil are confirmed by an in vivo assay in mice, the daily oral administration of methanolic extract (500 and 800 mg/kg/day) and fixed oil (2 and 4 ml/kg/day) during 21 days, resulted in a significant enhancement of the blood total antioxidant capacity (measured by KRL test) and the plasmatic antioxidant capacity towards DPPH radical.

Calculation of the Forces Acting on the Knee Joint When Rising from Kneeling Positions (Effects of the Leg Alignment and the Arm Assistance on the Knee Joint Forces)

Knee joint forces are available by in vivo measurement using an instrumented knee prosthesis for small to moderate knee flexion but not for high flexion yet. We created a 2D mathematical model of the lower limb incorporating several new features such as a patello-femoral mechanism, a thigh-calf contact at high knee flexion and co-contracting muscles' force ratio, then used it to determine knee joint forces arising from high knee flexions in four kneeling conditions: rising with legs in parallel, with one foot forward, with or without arm use. With arms used, the maximum values of knee joint force decreased to about 60% of those with arms not used. When rising with one foot forward, if arms are not used, the forward leg sustains a force as large as that sustained when rising with legs parallel.

Biocompatibility of NiTi Alloy Implants in vivo

In this study, the powders of Ni and Ti with 50.5 at.% Ni for 12 h were blended and cold pressed at the different pressures (50, 75 and100 MPa).The porous product obtained after Ni-Ti compacts were synthesized by SHS (self-propagating hightemperature synthesis) in the different preheating temperatures (200, 250 and 300oC) and heating rates (30, 60 and 90oC/min). The effects of the pressure, preheating temperature and heating rate were investigated on biocompatibility in vivo. The porosity in the synthesized products was in the range of 50.7–59.7 vol. %. The pressure, preheating temperature and heating rate were found to have an important effect on the biocompatibility in-vivo of the synthesized products. Max. fibrotic tissue within the porous implant was found in vivo periods (6 months), in which compacting pressure 100MPa.

Computer Modeling of Drug Distribution after Intravitreal Administration

Intravitreal injection (IVI) is the most common treatment for eye posterior segment diseases such as endopthalmitis, retinitis, age-related macular degeneration, diabetic retinopathy, uveitis, and retinal detachment. Most of the drugs used to treat vitreoretinal diseases, have a narrow concentration range in which they are effective, and may be toxic at higher concentrations. Therefore, it is critical to know the drug distribution within the eye following intravitreal injection. Having knowledge of drug distribution, ophthalmologists can decide on drug injection frequency while minimizing damage to tissues. The goal of this study was to develop a computer model to predict intraocular concentrations and pharmacokinetics of intravitreally injected drugs. A finite volume model was created to predict distribution of two drugs with different physiochemical properties in the rabbit eye. The model parameters were obtained from literature review. To validate this numeric model, the in vivo data of spatial concentration profile from the lens to the retina were compared with the numeric data. The difference was less than 5% between the numerical and experimental data. This validation provides strong support for the numerical methodology and associated assumptions of the current study.