Neuroblasts Micropatterning on Nanostructural Modified Chitosan Membranes

The study describes chitosan membrane platform modified with nanostructure pattern which using nanotechnology to fabricate. The cell-substrate interaction between neuro-2a neuroblasts cell lines and chitosan membrane (flat, nanostructure and nanostructure pattern types) was investigated. The adhered morphology of neuro-2a cells depends on the topography of chitosan surface. We have found that neuro-2a showed different morphogenesis when cells adhered on flat and nanostructure chitosan membrane. The cell projected area of neuro-2a on flat chitosan membrane is larger than on nanostructure chitosan membrane. In addition, neuro-2a cells preferred to adhere on flat chitosan surface region than on nanostructure chitosan membrane to immobilize and differentiation. The experiment suggests surface topography can be used as a critical mechanism to isolate group of neuro-2a to a particular rectangle area on chitosan membrane. Our finding will provide a platform to take patch clamp to record electrophysiological behavior about neurons in vitro in the future.

In vitro Plant Regeneration of Java Vetiver (Vetiveria zizanioides)

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

Effect of Ginger and L-Carnitine on the Reproductive Performance of Male Rats

In this study, we investigated the effects of ginger and L-carnitine on the reproductive performance of male rats with respect to semen parameters, male sex hormones and the testicular antioxidant system. A total of sixty mature male albino rats were divided into four groups of fifteen rats. The control group received saline, whereas the other three groups received ginger (100 mg kg-1 d- 1.), L-carnitine (150 mg kg-1 d-1.) or a combination of both ginger (100 mg kg-1 d-1.) and L-carnitine (150 mg kg-1 d-1.) via a stomach tube daily for one month. At the end of the treatment period, the rats were sacrificed, and their sperm characteristics (count, motility and viability), antioxidant enzyme factors levels (reduced glutathione, catalase, superoxide dismutase and total antioxidant capacity) and sex hormone levels (testosterone, Follicle stimulating hormone(FSH) and luteinizing hormone (LH) were analysed. Our results showed that the three experimental treatments improved sperm parameters, antioxidant enzyme activity and testosterone hormone levels; the most pronounced positive effects were observed in the group that received a combination of both ginger and L-carnitine. Therefore, the administration of a combination of ginger and L-carnitine may be beneficial for improving male sexual performance.

In Silico Analysis of Pax6 Interacting Proteins Indicates Missing Molecular Links in Development of Brain and Associated Disease

The PAX6, a transcription factor, is essential for the morphogenesis of the eyes, brain, pituitary and pancreatic islets. In rodents, the loss of Pax6 function leads to central nervous system defects, anophthalmia, and nasal hypoplasia. The haplo-insufficiency of Pax6 causes microphthalmia, aggression and other behavioral abnormalities. It is also required in brain patterning and neuronal plasticity. In human, heterozygous mutation of Pax6 causes loss of iris [aniridia], mental retardation and glucose intolerance. The 3- deletion in Pax6 leads to autism and aniridia. The phenotypes are variable in peneterance and expressivity. However, mechanism of function and interaction of PAX6 with other proteins during development and associated disease are not clear. It is intended to explore interactors of PAX6 to elucidated biology of PAX6 function in the tissues where it is expressed and also in the central regulatory pathway. This report describes In-silico approaches to explore interacting proteins of PAX6. The models show several possible proteins interacting with PAX6 like MITF, SIX3, SOX2, SOX3, IPO13, TRIM, and OGT. Since the Pax6 is a critical transcriptional regulator and master control gene of eye and brain development it might be interacting with other protein involved in morphogenesis [TGIF, TGF, Ras etc]. It is also presumed that matricelluar proteins [SPARC, thrombospondin-1 and osteonectin etc] are likely to interact during transport and processing of PAX6 and are somewhere its cascade. The proteins involved in cell survival and cell proliferation can also not be ignored.

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

Molecular Mechanism of Amino Acid Discrimination for the Editing Reaction of E.coli Leucyl-tRNA Synthetase

Certain tRNA synthetases have developed highly accurate molecular machinery to discriminate their cognate amino acids. Those aaRSs achieve their goal via editing reaction in the Connective Polypeptide 1 (CP1). Recently mutagenesis studies have revealed the critical importance of residues in the CP1 domain for editing activity and X-ray structures have shown binding mode of noncognate amino acids in the editing domain. To pursue molecular mechanism for amino acid discrimination, molecular modeling studies were performed. Our results suggest that aaRS bind the noncognate amino acid more tightly than the cognate one. Finally, by comparing binding conformations of the amino acids in three systems, the amino acid binding mode was elucidated and a discrimination mechanism proposed. The results strongly reveal that the conserved threonines are responsible for amino acid discrimination. This is achieved through side chain interactions between T252 and T247/T248 as well as between those threonines and the incoming amino acids.

Herpes Simplex Virus Type I Infection of Mice Testis and Effect on Fertility

The objective of current issue was to develop a model of testicular herpes simplex virus (HSV) type I infection for assessment of viral effect on fertility. 56 male mice were inoculated intraperitoneally with different concentrations of HSV on 8 day post partum. It was revealed that the optimal dose was 100 plaque forming units per mice as it provided testicular infection in 100% of survivors. HSV proteins were detected both in somatic and germ cells (spermatogonia, spermatocytes, spermatides). Although DNA load in testis was descending from 3 to 28 days post infection only 12.5% of infected males had offspring after mating with uninfected females comparing to 87.5% in control (p=0.012). These results are the first direct evidence for HSV impact in male sterility. Prepuberal mice appeared to be a suitable model for investigation of pathogenesis of virus-associated fertility disorders.

Visualisation Techniques Connecting VRML and GENESIS Environments

We created the tool, which combines the powerful GENESIS (GEneral NEural SImulation System) simulation language with the up-to-date visualisation and internet techniques. Our solution resides in the connection between the simulation output from GENESIS, which is converted to the data-structure suitable for WWW browsers and VRML (Virtual Reality Modelling Language) viewers. The selected GENESIS simulations are once exported into the VRML code, and stored in our neurovisualisation portal (webserver). There, the loaded models, demonstrating mainly the spread of electrical signal (action potentials, postsynaptic potentials) along the neuronal membrane (axon, dendritic tree, neuron) could be displayed in the client-s VRML viewer, without interacting with original GENESIS environment. This enables the visualisation of basic neurophysiological phenomena designed for GENESIS simulator on the independent OS (operation system).

Computational Design of Inhibitory Agents of BMP-Noggin Interaction to Promote Osteogenesis

Bone growth factors, such as Bone Morphogenic Protein-2 (BMP-2) have been approved by the FDA to replace grafting for some surgical interventions, but the high dose requirement limits its use in patients. Noggin, an extracellular protein, blocks the effect of BMP-2 by binding to BMP. Preventing the BMP-2/noggin interaction will help increase the free concentration of BMP-2 and therefore should enhance its efficacy to induce bone formation. The work presented here involves computational design of novel small molecule inhibitory agents of BMP-2/noggin interaction, based on our current understanding of BMP-2, and its known putative ligands (receptors and antagonists). A successful acquisition of such an inhibitory agent of BMP-2/noggin interaction would allow clinicians to reduce the dose required of BMP-2 protein in clinical applications to promote osteogenesis. The available crystal structures of the BMPs, its receptors, and the binding partner noggin were analyzed to identify the critical residues involved in their interaction. In presenting this study, LUDI de novo design method was utilized to perform virtual screening of a large number of compounds from a commercially available library against the binding sites of noggin to identify the lead chemical compounds that could potentially block BMP-noggin interaction with a high specificity.

Gene Expressions Associated with Ultrastructural Changes in Vascular Endothelium of Atherosclerotic Lesion

Attachment of the circulating monocytes to the endothelium is the earliest detectable events during formation of atherosclerosis. The adhesion molecules, chemokines and matrix proteases genes were identified to be expressed in atherogenesis. Expressions of these genes may influence structural integrity of the luminal endothelium. The aim of this study is to relate changes in the ultrastructural morphology of the aortic luminal surface and gene expressions of the endothelial surface, chemokine and MMP-12 in normal and hypercholesterolemic rabbits. Luminal endothelial surface from rabbit aortic tissue was examined by scanning electron microscopy (SEM) using low vacuum mode to ascertain ultrastructural changes in development of atherosclerotic lesion. Gene expression of adhesion molecules, MCP-1 and MMP-12 were studied by Real-time PCR. Ultrastructural observations of the aortic luminal surface exhibited changes from normal regular smooth intact endothelium to irregular luminal surface including marked globular appearance and ruptures of the membrane layer. Real-time PCR demonstrated differentially expressed of studied genes in atherosclerotic tissues. The appearance of ultrastructural changes in aortic tissue of hypercholesterolemic rabbits is suggested to have relation with underlying changes of endothelial surface molecules, chemokine and MMP-12 gene expressions.

Gene Selection Guided by Feature Interdependence

Cancers could normally be marked by a number of differentially expressed genes which show enormous potential as biomarkers for a certain disease. Recent years, cancer classification based on the investigation of gene expression profiles derived by high-throughput microarrays has widely been used. The selection of discriminative genes is, therefore, an essential preprocess step in carcinogenesis studies. In this paper, we have proposed a novel gene selector using information-theoretic measures for biological discovery. This multivariate filter is a four-stage framework through the analyses of feature relevance, feature interdependence, feature redundancy-dependence and subset rankings, and having been examined on the colon cancer data set. Our experimental result show that the proposed method outperformed other information theorem based filters in all aspect of classification errors and classification performance.

A Novel Nucleus-Based Classifier for Discrimination of Osteoclasts and Mesenchymal Precursor Cells in Mouse Bone Marrow Cultures

Bone remodeling occurs by the balanced action of bone resorbing osteoclasts (OC) and bone-building osteoblasts. Increased bone resorption by excessive OC activity contributes to malignant and non-malignant diseases including osteoporosis. To study OC differentiation and function, OC formed in in vitro cultures are currently counted manually, a tedious procedure which is prone to inter-observer differences. Aiming for an automated OC-quantification system, classification of OC and precursor cells was done on fluorescence microscope images based on the distinct appearance of fluorescent nuclei. Following ellipse fitting to nuclei, a combination of eight features enabled clustering of OC and precursor cell nuclei. After evaluating different machine-learning techniques, LOGREG achieved 74% correctly classified OC and precursor cell nuclei, outperforming human experts (best expert: 55%). In combination with the automated detection of total cell areas, this system allows to measure various cell parameters and most importantly to quantify proteins involved in osteoclastogenesis.