Synapses are contact points where one neuron communicates with another. The morphological change of synapses under various physiological or pathological conditions has long been hypothesized to modify their functional properties. 3-dimensional (3-D) reconstruction of synapses with serial ultrathin sections has contributed to the understanding of ultrastructural dimensions and compositions of synapses. The 3-D reconstruction procedures, however, require a great amount of expertise as well as include prohibitively timeconsuming processes. Here, we introduce efficient 3-D reconstruction technique using high-voltage electron microscopy (HVEM). Primarily, we established an optimal section thickness and staining condition to observe synaptic structures in detail under HVEM. The result showed that synaptic profiles were preserved at the section thickness of 250 nm without the overlapping of synaptic ultrastructures. An increase in the reaction time of en bloc staining was most efficient to enhance contrast than the extension of postembedding staining or the addition of uranyl acetate during dehydration. Then, 3-D reconstruction of parallel fiber-Purkinje cell synapses in the rat cerebellum was carried out with serial HVEM images and reconstruction software. The images were aligned and the contours of synapses were outlined on each section. 3-D synapses were finally extracted from the section files by grouping all the synaptic contours. The reconstructed synapse model clearly demonstrated the configuration of pre and postsynaptic components. These results suggest that 3-D reconstruction of synapses using HVEM is much efficient and suitable for massive quantitative studies on synaptic connectivity than conventional TEM approach using numerous ultrathin sections.
Present study was performed to observe the tegumental ultrastructures by the developmental stages which derived from the experimental life cycle of Spirometra erinacei in laboratory conditions. In SEM view, coracidium was spherical in shape with numerous cilia, and its surface was covered with long cilia, tuberclelike projections with millet-like processes, and small holes. The body surface of procercoid was covered with numerous pointed microtriches except that of frontal pit with stout spine-like ones. However that of cercomer was covered with somewhat sparse blunt-tiped microtriches. Plerocercoids of 3 days old resembled the mature procercoid in shape, and their frontal pits were covered with numerous stout spine-like microtriches. However frontal pit and body surface in more than 5 days old ones were covered with conoid microtriches. On the surface of adult scolex, hairly long filamentous and stout short microtriches were mixedly distributed. Filamentous microtriches were more densely distributed in the anterior portion than in the posterior of scolex. The neck and immature proglottid were covered with only stout short conoid microtriches. In TEM view of coracidia, embryophore and oncosphere were obviously distinguished. The embryophore contained numerous glycogen particles, mitochondria and lipid granules. The cilia on the surface of embryophore rooted in the coracidial sheath, and consisted of 9 pairs of microtubules and 2 core complex. The oncosphere was covered with a thin and unarmed tegument, and was multi-nucleated. The protoplasmic layer of procercoid and plerocercoid consisted of disc-shaped bodies, vacuoles and mitochondria. Their tegumental cells commonly retained a nucleus, granular endoplasmic reticulums and secretory granules. The protoplasmic layer of plerocercoid was more compacted than that of procercoid. From the above results, it was confirmed that the tegumental ultrastructures are something different according to the developmental stages of S. erinacei.
Bo-Young Choe;Sei-Kwon Kang;Myoung-Ja Chu;Hyun-Man Baik;Euy-Neyng Kim
Investigative Magnetic Resonance Imaging
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v.5
no.2
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pp.138-148
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2001
Purpose : Within a clinically acceptable time frame, we obtained the high resolution MR images of the human brain, knee, foot and wrist from 3T whole-body MRI system which was equipped with the world first 37 active shield magnet. Materials and Methods : Spin echo (SE) and Fast Spin Echo (FSE) images were obtained from the human brain, knee, foot and wrist of normal subjects using a homemade birdcage and transverse electromagnetic (TEM) resonators operating in quadrature and tuned to 128 MHz. For acquisition of MR images of knee, foot and wrist, we employed a homemade saddle shaped RF coil. Topical common acquisition parameters were as follows: matrix=$512{\times}512$, field of view (FOV) =20 cm, slice thickness = 3 mm, number of excitations (NEX)=1. For T1-weighted MR images, we used TR = 500 ms, TE = 10 or 17.4 ms. For T2-weighted MR images, we used TR=4000 ms, TE = 108 ms. Results : Signal to noise ratio (SNR) of 3T system was measured 2.7 times greater than that of prevalent 1.5T system. MR images obtained from 3T system revealed numerous small venous structures throughout the image plane and provided reasonable delineation between gray and white matter. Conclusion The present results demonstrate that the MR images from 3T system could provide better diagnostic quali\ulcorner of resolution and sensitivity than those of 1.5T system. The elevated SNR observed in the 3T high field magnetic resonance imaging can be utilized to acquire images with a level of resolution approaching the microscopic structural level under in vivo conditions. These images represent a significant advance in our ability to examine small anatomical features with noninvasive imaging methods.
This study was conducted to determine the antiseptic effect of stabilized chlorine dioxide (S-$ClO_2$) on muscle tissue of rats. Skeletal muscle of 8-week old Sprague-Dawley rats was used. Light and transmission electron microscopic findings were observed in the control group, which was not treated with stabilized chlorine dioxide, and in the experimental group, which was treated with a stabilized chlorine dioxide powder in aqueous solution. According to the LM and TEM observations, the day 1 control group showed the initiation of endomysium collapse resulting in an unclear boundary of muscle fibers, and partial collapse of the mitochondrial membranes. All endomysium had collapsed, and bacteria were observed among muscle fibers in the day 2 and later groups. Shapes of muscles were not distinguishable in day 3 or later groups. In contrast, the day 1 and 3 experimental groups revealed detailed structure of typical muscles, but partial collapse of the mitochondrial membranes was observed in the day 3 and later groups. Subsequently, connective tissues collapsed and structures in the shape of concentric circles were observed. In summary, the day 1 control group showed the initial collapse of tissues, and shapes were not distinguishable in the day 3 and later groups because most of the tissues had collapsed. In contrast, the day 3 experimental group showed partial collapse, but the overall shapes of muscles were maintained as time went on, confirming the antiseptic effect of stabilized chlorine dioxide on muscles.
For veterinary imaging diagnosis, we obtained MR images of the canine brain, spine, kidney and pelvis from 3T MRI system which was equipped with the world first 3T active shield magnet. Spin echo (SE) and fast Spin Echo (FSE) images were obtained from the canine brain, spine, kidney and pelvis of normal and sick dogs using a homemade birdcage and transverse electromagnetic (TEM) resonators operating in quadrature and tuned to 128 MHz. In addition, we employed a homemade saddle shaped RF coil. Typical common acquisition parameters were as follows: matrix=512$\times$512, field of view (FOV)=20cm, slice thickness=3 w, number of excitations (NEX)=1. For T1-weighted MR images, we used TR=500 ms, TE=10 or 17.4 ms. For T2-weighted MR images, we used TR=4000 ms, TE=108 ms. Signal to noise ratio (SNR) of 3T system was measured 2.7 times greater than that of prevalent 1.57 system. The high resolution images acquired in this study represent more than a 4-fold increase in in-plane resolution relative to conventional images obtained with a 20 cm field of view and a 5 mm slice thickness. MR images obtained from 3T system revealed numerous small venous structures throughout the image plane and provided reasonable delineation between gray and white matter The present results demonstrate that the MR images from 3T system could provide better diagnostic quality of resolution and sensitivity than those of 1.5T system. The elevated SNR observed in the 3T high field magnetic resonance imaging can be utilized to acquire images with a level of resolution approaching the microscopic structural level under in vivo conditions. These images represent a significant advance in our ability to examine small anatomical features with noninvasive imaging methods. Moreover, MRI technique could begin to apply for veterinary medicine in Korea.
Even traces of CO in the hydrogen-rich feed gas to proton exchange membrane fuel cells (PEMFC) poison the platinum anode electrode and dramatically decrease the power output. In this work, a variety of catalytic materials consisting of $Cu/Ce_xZr_{1-x}O_2$, (x = 0.0-1.0) were synthesised, characterized and tested for CO oxidation and preferential oxidation of CO (PROX). These catalysts prepared by hydrothermal and deposition-precipitation methods. The catalysts were characterized by XRD, XRF, SEM, BET, $N_2O$ titration and oxygen storage capacity (OSC) measurement. The effects of composition of the support and degree of excess oxygen were investigated fur activity and $CO_2$ selectivity with different temperatures. The composition of the support markedly influenced the PROX activity. Among the various $Cu/Ce_xZr_{1-x}O_2$ catalysts having different composition, $Cu/Ce_{0.9}Zr_{0.1}O_2$ and $Cu/Ce_{0.7}Zr_{0.3}O_2$ showed the highest activities (>99%) and selectivities (ca.50%) in the temperature range of $150{\sim}160^{\circ}C$. It was found that by using of $Ce_xZr_{1-x}O_2$ mixed oxide support which possesses a high oxygen storage capacity, oxidation-reduction activity of Cu-based catalyst was improved, which resulted in the increase of catalytic activity and selectivity of CO oxidation in excess $H_2$ environments.
Park, Young-Hee;Ahn, E-Tay;Ko, Jeong-Sik;Park, Dae-Kyoon;Kim, Myeong-Soo;Park, Kyung-Ho
Applied Microscopy
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v.37
no.1
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pp.23-33
/
2007
The lacrimal gland are compound tubule-acinar glands. The main lacrimal function is the production of the aqueous layer, the thickest and major constituent of the precorneal tear film. The lacrimal gland also has an important function in the defense system of the ocular surface, forming a part of the conjunctival-associated hymphoid tissue. The ultrastructural characteristics of the lacrimal gland of the rabbit were described. The lacrimal tissues of rabbits were processed through the conventional techniques for transmission electron microscopy. The secretory portions consisted of three cell types: 1. Serous cells with electron dense secretory granules. 2. Seromucous cells containing variable moderately electron dense secretory granules with flocculent material. 3. Mucous rolls containing mucous secretory granules. The serous cells were situated at the basal portion of acini, and they contained electron dense granules of variable densities and sizes. The seromucous cells contained a few protein secretory granules and more mucous secretory granules. The mucous cells contained even fewer protein secretory granules and exclusively mucous secretory granules. The epithelium of the intralobular ducts showed secretory granules, junctional complexes, and large basolateral intercellular spaces with lateral folds. These study might be helpful in determining inter-relationships, similarities and differences among the orbital glands of various physiological or pathological conditions.
Plants of Vitex negundo are known to develop numerous trichomes throughout their body, where certain trichome types have been believed to be one of the plausible structures for the unique scents. In the current study. structural aspects of the trichomes have been examined in leaves and stems of Vitex negundo using TEM and SEM. Trichome types as well as structural changes that occurred in certain trichomes during secretion have been mainly focused. Three type of glandular trichomes and two types of non-glandular trichomes were developed in the epidermis of young and mature Vitex negundo plants. The glandular trichomes included the peltate type (Type 1), the capitate type (Type 2), and degraded capitate type (Type 3), whereas the non-glandular warty trichomes contained the multicellular (Types 4) and unicellular type (Type 5). Type 1 and 2 consisted of head and stalk cells, but their number and size were different. One secretory cavity was formed from the four head cells in the former, but only two head cells were involved in the latter. The cytoplasmic density in the head cell was quite high and in particular, sER and Golgi bodies were well developed. At initiation of their development, the cuticle layer of the head cells separated from the outer tangential wall to form a secretory cavity. Subsequently the cavity expanded acropetally and a large number of secretory vesicles continuously produced from the head cells until they filled the entire cavity. The cavity contained materials that would be soon discharged into intercellular spaces and/or into the air. The cavity began to decrease the volume by contracting at initial secretion but degrade rapidly within short time. It has been suggested that the mode of secretion in V. negundo is probably the eccrine secretion, since no break or rupture of the cavity has been observed during examination. Contrastingly Type 3 exhibited deterioration of the head cell at early stage. Type 4 was about $110{\sim}190{\mu}m$ long, consisting of $2{\sim}3$ cells, and distributed more in the adaxial epidermis compared to the abaxial surface. However, $20{\sim}30{\mu}m$ long Type 5 was extremely dense in both epidermis. Among several trichome types, Type 1 and 2 probably play an important role in discharging unique aromatic scents in plants of V. negundo.
This study was conducted to the effect of temporary cement on the adhesiveness of dentin bonding agent to dentin surface. One hundred freshly extracted bovine mandibular incisors were grinded to expose flat labial dentin surface. The dentin surfaces were temporarized with either eugenol-containing temporary cement(TemBond and Zinc Oxide Eugenol cement) or non-eugenol temporary cement(Nogenol and TempBond NE) for 7days, and then the temporarization was removed with surgical currette and the exposed dentin surfaces were water-rinsed. Bonding specimens were made by use of All-Bond 2 and Super-Bond C&B dentin bonding agent, and stored in $37^{\circ}C$ distilled water for 24hours. The tensile bond strenth and the cohesive failure rate were measured, and then the pretreated dentin surfaces which the temporary cement had been applied to and removed from and the fractured dentin surfaces after bonding test were examined under scanning electron microscope. The results were as follows : In case of bonding with All-Bond 2, tensile bond strength of each experimental group was lower than that of the control group(p<0.05), but there was no significant difference between the bond strengths of the control group and each experimental group in case of bonding with Super-Bond C&B(p>0.05). No significant difference between tensile bond strength of experimental group, whether temporary cement contains eugenol or not, was seen(p>0.05). In case of bonding with All-Bond 2, the control group showed cohesive-adhesive mixed failure mode and the experimental groups mainly showed adhesive failure mode, but in case of bonding with Super-Bond C&B, almost of the control and the experimental groups mainly showed cohesive failure mode. On SEM examination, all of the dentin specimens pretreated with either 10 % phosphoric acid or 10% citric acid after application of the temporary cements demonstrated remnants of temporary cement attached to dentin surface.
Kim, Myeong-Su;Ohn, Young-Seok;Lee, Kwang-Won;Son, Ho-Hyun
Restorative Dentistry and Endodontics
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v.23
no.1
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pp.141-161
/
1998
The depth and patterns of demineralization according to the difference in concentration and application time of phosphoric acid were observed through the transmission electron microscope, and shear bond strengths to the acid -conditioned dentin were then measured and compared with the TEM results. To investigate the influence of polymer addition into the phosphoric acid and the effect of difference in concentration and application time of the acid, the specimens were randomly divided into 9 groups. Among the specimens, the exposed dentin surfaces were acid-conditioned with 10% polymer-thickened phosphoric acid(All Bond 2, Bisco, U.S.A.) and aqueous 10%, 20%, 30%, 40% phosphoric acid for 20 seconds, The rest of the specimens were acid-conditioned with 10% phosphoric acid for 15s, 30s, 60s, 120s respectively. The specimens were immersed in 4% glutaraldehyde in 0.1M sodium cacodylate buffer and postfixed with 1 % osmium tetroxide without decalcification and then observed under a JEOL Transmission Electron Microscope(JEM 1200 EX II, Japan). After the specimens were acid-conditioned as the above, primer and adhesive resin were applied to blot-dried dentin and shear bond strengths were then measured and analysed. The results were as follows : 1. The intertubular demineralization depth of 4.0-$5.0{\mu}m$ in 10% polymer-thickened phosphoric acid gels was similar or slightly deeper than that of 4.0-$4.5{\mu}m$ in aqueous 10% phosphoric acid solution. 2. The intertubular demineralization depth of aqueous 20%, 30% and 40% phosphoric acid solution was 6.5-$7.0{\mu}m$, 6.5-$7.5{\mu}m$ and 9.0-$15.0{\mu}m$ respectively. It showed that the depth of dentin demineralization is partly related to the concentration of phosphoric acid solution. 3. The intertubular demineralization depth of aqueous 10% phosphoric acid solution in application time for 15s, 30s, 60s and 120s was 2.5-$3.0{\mu}m$, 4.0-$6.0{\mu}m$, 6.5-$7.0{\mu}m$ and 8.5-$14.0{\mu}m$ respectively. It showed that the depth of dentin demineralization is directly related to the application time of phosphoric acid solution. 4. The partially demineralized dentin layer between demineralized collagen layer and unaffected dentin was showed to a width of 0.5-$1.0{\mu}m$ in lower concentration groups treated with aqueous 10% phosphoric acid for 20s, 60s, 120s and 20% phosphoric acid for 20s. 5. The demineralization effect at the border of intertubular-peritubular junction was less evident than that in the peritubular and intertubular dentin. The collagen fibers in the intertubular dentin had a random orientation, whereas those that lined the tubules were circumferentially aligned. The cross-linkage of dentinal collagen in demineralized collagen layer was clearly seen. 6. A statistically significant difference of bond strengths according to the difference in phosphoric acid concentration did not exist among the groups treated with 10%, 20%, 30% and 40% acid solution (P>0.05). However, bond strengths to the treated dentin with 10% phosphoric acid solution for 30s were significantly higher than that for 120s (P<0.05).
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