Scanning Electron Microscopic Observation of the Crista Ampullaris

The crista ampullaris of the guinea pig and the bull frog were investigated by scanning electron microscopy. The crista ampullaris were freeze fractured or sheared followed by maceration with 0.1% OsO4 solution. Following this, three-dimensional intracellular structures were observed. The mitochondria of the sensory cells varied in shape from globular to long and slender. Golgi apparatus and endoplasmic reticulum of the sensory cells were also demonstrated clearly. Nerve elements, nerve endings and synaptic structures were also observed stereoscopically

Intracellular Structure of the Outer Hair Cell of the Organ of Corti

The intracellular structure of the outer hair cells of the normal guinea pig organ of Corti was investigated three dimensionally by scanning electron microscope. Freeze fracturing technique followed by maceration with a 0.1% OsO4 solution (osmic maceration method) was used. Among the cell organelles, the endoplasmic reticulum (ER) showed the most interesting features, such as subsurface cisternae and lamellar bodies. The subsurface cisterna which formed a stratiform network covered the inner surface of the cell membrane and the stratiform structure disappeared at the infranuclear region. Variously shaped mitochondria (spherical, cylindrical and branched) were found on the innermost layer of the subsurface cisterna. The lamellar body which was located beneath the cuticular plate consisted of dilated cisternae and tubular ER and was surrounded by mitochondria. The tubular ER of the lamellar body were continuous with the subsurface cisterna

The Morphological Changes in the Vestibular Sensory Epithelia Following Electrical Stimulation

The morphological changes of the vestibular sensory epithelia of the guinea pig following electrical stimulation were investigated using scanning electron microscope. Positive and negative square wave pulse stimulation was given through a silver ball electrode placed on the round window membrane for one hour. The current intensities used were 100, 200 and 300 A. While the direct current stimulation at intensities of 100 or 200 A did not cause any significant changes, severe damage of the utricular macula and the ampullar crista of the lateral semicircular canal was observed at 300 A. The degenerative changes such as fusion of sensory hairs, protrusion of the cuticular plate and loss of sensory cells were found on both the utricle and the semicircular canal. In the most severely damaged area, the sensory epithelial surface was badly torn apart. In the clinical application of direct current to the inner ear for relieving tinnitus, special attention should be paid to the vestibular organ

Study on the behavior of small droplet impinging onto a hot surface

This paper was presented at the 3rd Micro and Nano Flows Conference (MNF2011), which was held at the Makedonia Palace Hotel, Thessaloniki in Greece. The conference was organised by Brunel University and supported by the Italian Union of Thermofluiddynamics, Aristotle University of Thessaloniki, University of Thessaly, IPEM, the Process Intensification Network, the Institution of Mechanical Engineers, the Heat Transfer Society, HEXAG - the Heat Exchange Action Group, and the Energy Institute.The effects of droplet diameter, surface roughness, and impinging velocity on the behavior of droplet impinging onto a hot surface have been studied. The surface samples used in the experiment were cylinder blocks of stainless steel having four different degrees of roughness, i.e., Ra 0.04, 0.2, 3, and 10. The diameter and impinging velocity were controlled independently by using a micro-jet dispenser. Their values were in the ranges of 300–700 μm and 1.0–4.0 m/s, respectively. The contact time was found to increase with an increase in the surface roughness and was of the order of the self-oscillation of the water droplet. The maximum spread of droplet decreased with increasing impinging velocity. The cooling curve was obtained for the range of surface temperatures from 500 oC to 100 oC, and it was found that the cooling time decreased with an increase in the surface roughness of stainless steel. Moreover, the cooling effectiveness of each droplet increased with an increase in the surface roughness.This study was supported by the Grant-in-Aid for Scientific Research (A) 21246036 from MEXT

Morphological Changes in the Vestibular Epithelia and Ganglion Induced by Ototoxic Drug

The morphological changes of the vestibular sensory epithelia and the vestibular ganglions induced by Gentamicin (GM) were investigated using scanning electron microscope, transmission electron microscope and light microscope. The guinea pigs were injected with a single application of 4 mg (0.1ml) of GM into the middle ear through the tympanic membrane. The vestibular organs and the ganglions were observed up to 6 months after the treatment. Four days after the injection, fused, ballooned and missing cilia were observed in the vestibular sensory epithelia. These changes progressed and extended toward the periphery of the crista and the macula. The changes of the vestibular ganglions were first observed one month after the treatment. The degenerative process started from destruction of the mitochondrial cristae and vacuolization of the cytoplasm in the Schwann cell. The next step of the change was dissociation of the myelin sheath around the ganglion cell. The cytoplasmic organelles in the ganglion cell gradually deteriorated. At the later stage, the myelin sheath around the ganglion cell disappeared and the number of the cell reduced. Furthermore, the myelin sheath of the nerve fiber was dissociated. In this study the signs of the vestibular ganglion damage were later than that of the vestibular organ. However, we thought the changes in the ganglion are probably due to direct influence of GM, since the degeneration was found to develop in a relatively short period

Agent based modelling helps in understanding the rules by which fibroblasts support keratinocyte colony formation

Background: Autologous keratincoytes are routinely expanded using irradiated mouse fibroblasts and bovine serum for clinical use. With growing concerns about the safety of these xenobiotic materials, it is desirable to culture keratinocytes in media without animal derived products. An improved understanding of epithelial/mesenchymal interactions could assist in this. Methodology/Principal Findings: A keratincyte/fibroblast o-culture model was developed by extending an agent-based keratinocyte colony formation model to include the response of keratinocytes to both fibroblasts and serum. The model was validated by comparison of the in virtuo and in vitro multicellular behaviour of keratinocytes and fibroblasts in single and co-culture in Greens medium. To test the robustness of the model, several properties of the fibroblasts were changed to investigate their influence on the multicellular morphogenesis of keratinocyes and fibroblasts. The model was then used to generate hypotheses to explore the interactions of both proliferative and growth arrested fibroblasts with keratinocytes. The key predictions arising from the model which were confirmed by in vitro experiments were that 1) the ratio of fibroblasts to keratinocytes would critically influence keratinocyte colony expansion, 2) this ratio needed to be optimum at the beginning of the co-culture, 3) proliferative fibroblasts would be more effective than irradiated cells in expanding keratinocytes and 4) in the presence of an adequate number of fibroblasts, keratinocyte expansion would be independent of serum. Conclusions: A closely associated computational and biological approach is a powerful tool for understanding complex biological systems such as the interactions between keratinocytes and fibroblasts. The key outcome of this study is the finding that the early addition of a critical ratio of proliferative fibroblasts can give rapid keratinocyte expansion without the use of irradiated mouse fibroblasts and bovine serum

Effects of Payena dasyphylla (Miq.) on hyaluronidase enzyme activity and metalloproteinases protein expressions in interleukin-1beta stimulated human chondrocytes cells

Background: Hyaluronidases have been found as the target enzymes in the development of osteoarthritis (OA) disease. While there is still no curative treatment for this disease, recent studies on the treatment of OA were focused on the effectiveness of natural products which are expected to improve the symptoms with minimal side effects. The aim of this study was to screen selected Malaysian plants on their anti-hyaluronidase activity as well as to evaluate the active plant and its derived fractions on its potential anti-arthritic and antioxidant activities.Methods: A total of 20 methanolic crude extracts (bark and leaf) from ten different plants were screened using a colorimetric hyaluronidase enzymatic assay. The active plant extract (Payena dasyphylla) was then studied for its hyaluronidase inhibitory activity in the interleukin-1β (IL-1β) stimulated human chondrocytes cell line (NHAC-kn) using zymography method. The Payena dasyphylla methanolic bark extract was then fractionated into several fractions in where the ethyl acetate (EA) fraction was evaluated for its inhibitory effects on the HYAL1 and HYAL2 gene expressions using reverse transcription-polymerase chain reaction (RT-PCR) technique. While the MMP-3 and MMP-13 protein expressions were evaluated using western blot method. The phenolic and flavonoid contents of the three fractions as well as the antioxidant property of the EA fraction were also evaluated.Results: Bark extract of Payena dasyphylla (100 μg/ml) showed the highest inhibitory activity against bovine testicular hyaluronidase with 91.63%. The plant extract also inhibited hyaluronidase expression in the cultured human chondrocyte cells in response to IL-1β (100 ng/ml). Similarly, treatment with Payena dasyphylla ethyl acetate (EA) fraction (100 μg/ml) inhibited the HYAL1 and HYAL2 mRNA gene expressions as well as MMP-3 and MMP-13 protein expression in a dose dependent manner. Payena dasyphylla EA fraction has demonstrated the highest amount of phenolic and flavonoid content with 168.62 ± 10.93 mg GAE/g and 95.96 ± 2.96 mg RE/g respectively as compared to water and hexane fractions. In addition, the Payena dasyphylla EA fraction showed strong antioxidant activity with IC50 value of 11.64 ± 1.69 μg/mL.Conclusion: These findings have shown that Payena dasyphylla might contained potential phenolic compounds that inhibiting the key enzyme in osteoarthritis development, which is the hyaluronidase enzyme through interruption of HYAL1 and HYAL1 gene expressions. The degradation of cartilage could also be inhibited by the plant through suppression of MMP-3 and MMP-13 protein expressions. We also reported that the inhibitory effect of Payena dasyphylla on hyaluronidase activity and expression might be due to its anti-oxidant property