• Clinical and Experimental Reproductive Medicine
• /
• v.24 no.3
• /
• pp.369-375
• /
• 1997

Studies were conducted to determine the efficiency of decondensation protocols. Sperm obtained from seven normal donors was immediately washed after liquefaction and then decondensed using the method of West et al. (1989) and my original protocol. My optimized protocol entailed mixing 1 ml aliquots of semen with 4 ml phosphate buffered saline (PBS). Following centrifugation, pellets were resuspended in 1 ml PBS containing 6 mM EDTA. After centrifugation, pellets were resuspended in 1 ml PBS containing 2 mM dithiothreitol at $37^{\circ}C$ for 45 min. Following mixing with 2 ml PBS and centrifugation, pellets were resuspended by vortexing. While vortexing, 5 ml of fixative were gently added. Slide preparation was accomplished using the smear method and it was stored at $4^{\circ}C$. When comparing these protocols, the degree of sperm decondensation and head swelling was monitored by measuring nuclear length, area, perimeter, and degree of roundness using FISH analysis software. Apparent copy number for chromosome 1 and, separately, for the sex chromosomes was determined by FISH using satellite DNA probes for loci DIZ1, DXZ1 and DYZ3. Sperm treated by my decondensation protocol showed significant increases (p<0.05) in length, area, perimeter, and degree of roundness. There was a significant decrease (p<0.05) in the frequency of nuclei displaying no signal but no change in the frequency of nuclei with two signals in samples decondensed by my protocol. My data suggested that decondensation using my original protocol may lower the frequency of cells with spurious "nullisomy" due to hybridization failure without inducing spurious "disomy" resulting from increased distances between split signals.

• Applied Microscopy
• /
• v.23 no.2
• /
• pp.41-52
• /
• 1993

This study was designed to observe the ultrastructural characteristics of hepatocytes, endothelial cells, Kupffer cells and Ito cells in the liver of Suncus Murinus. The specimens were processed for electron microscopy, following the immersing in the Karnovsky fixative for 8 hrs and 1% osmium tetroxide for 2 hrs. Hepatocytes contained large number of lipid droplets and large mitochondria. Spaces between the hepatocytes were narrow and in some area were irregular in contours with long and slender microvilli of the hepatocytes. Endothelial cells may contain lysosomes as Kupffer cells do. Bile canaliculi were relatively wide and easily seen in the lobule. Bile ductules located in the space of Disse were formed by the hepatocytes and small cells with or without lipid droplets. The results suggest that the Suncus Murinus liver shows some ultrastructural characteristics of aquatic animal livers, endothelial and Kupffer cells may be the same one only in the different functional states, and the bile ductule may be partly formed by the Ito cell.

• Journal of Korean Ophthalmic Optics Society
• /
• v.13 no.4
• /
• pp.135-139
• /
• 2008

Purpose: This study was designed to compare the accommodative amplitude whose 40s initial presbyopia patients divided into five occupation (driver, official worker, housewife, field worker, teacher). Methods: Both "push-up"and "inus lens to blur technique" methods were used to examine the average of accommodative amplitude. Results: All the average of accommodative amplitude (OD, OS, and OU) were the lowest in official worker group (3.27${\pm}$0.21D, 3.31${\pm}$0.22D, 3.54${\pm}$0.28D) and the highest in housewife group (4.07${\pm}$0.35D, 4.11${\pm}$0.35D, 4.37${\pm}$0.39D). Conclusions: Because occupational specifications including a fixative habit are able to change the binocular accommodation, it is important to understand thoroughly the patient's occupation when opticians make near vision prescription for initial presbyopia.

• Journal of the Korean Association of Oral and Maxillofacial Surgeons
• /
• v.47 no.1
• /
• pp.40-46
• /
• 2021

Marsupialization is widely used as a primary treatment modality for reducing size of large cysts. However, there is no recommendation for specific duration of marsupialization. In addition, Carnoy's solution usually is applied at the time of enucleation as a fixative agent. In this report, we present an appropriate marsupialization duration of ameloblastoma involving two unerupted teeth. In this present study, marsupialization using a Foley catheter was performed in two cases of ameloblastoma of the mandible involving two adjacent impacted teeth. Carnoy's solution was applied for 3-5 minutes after enucleation in both patients. Periodically during marsupialization, the size of the radiolucency was measured in panoramic view, and clinical examination was performed. No remarkable paresthesia or soft tissue injury was observed after application of Carnoy's solution or during follow-up. We recommend 12 to 16 weeks as an adequate marsupialization duration for a large ameloblastoma involving two impacted teeth based on increased radiopacity along the margins of the lesions. Poor oral hygiene was an issue after 12 weeks of marsupialization in one case. There were no remarkable complications with Carnoy's solution in either case. The Foley tube has a two-way system that is more effective for irrigating the cavity than is the conventional one-way system.

• Applied Microscopy
• /
• v.16 no.2
• /
• pp.1-13
• /
• 1986

The purpose of this study was to investigate the morphology and intercellular junctions of the odontoblast of dental pulp in the rat incisor by means of the freeze fracture electron microscopy. Twenty male Sprague-Dawley rats weighing $150{\sim}200g$ were used. After being anesthetized by an intraperitoneal injection of 0.5 ml sodium pentobarbital per kg in body weight(60 mg/ml) the animals were perfused with 2.5% glutaraldehyde-2% paraformaldehyde fixative in 0.1 M cacodylate buffer, pH 7.2 through the ascending aorta for one hour. The incisors were carefully extracted from the jaws and demineralized by suspending them in 0.1 M EDTA in 3% glutaraldehyde (pH 7.2) for two weeks. After demineralization, the specimens were obtained from the portion divided into five equal parts. For freeze-fracture replication, demineralized tissues were infiltrated for several hours with 10%, 25% glycerol in 0.1M cacodylate buffer as a cryoprotectant and then frozen in liquid Freon 22 and stored in liquid nitrogen. Fracturing and replication were done in Balzers BAF 400D high-vacuum freeze-fracture apparatus at $-120^{\circ}C$ under routine $5X10^{-7}$ Torr vacuum. The tissue was immediately replicated with platinum unidirectionally at $45^{\circ}$ angle and reinforced with carbon at $90^{\circ}$ angle unidirectionally or by using a rotary stage. The replication process was monitored by a quartz-crystal device. The replicas were immersed in 100% methanol overnight. The tissue was then digested from the replica by clorox (laundry bleach), placed into 5% EDTA, and washed repeatedly with distilled water. The replicas were picked up on 0.3% formvar-coated 75 mesh grids and examined in the JEOL 100B electron microscope. The results were as follows; 1. Both in thin sections and freeze-fracture replicas, three types of intercellular junctions were recognizable in the plasma membrane of odontoblast: gap junction, tight junction and desmosome-like junction. 2. The nuclear pores were evenly distributed over the nuclear envelope. The pore complex formed a ring about 70 nm in diameter. 3. Gap junctions were found between odontoblasts as well as odontoblasts and neighbouring pulp cells (fibroblast, subodontoblastic cell process, nerve-like fibre). Gap junctions, which were round, ellipsoid and pear-shaped and 600 nm in diameter, were observed in the odontoblast. 4. Numerous round and ellipsoid gap junctions could be frequently seen on the plasma membranes in cell body and apical part of the odontoblasts. On the P face, the junctions were recognized as a cluster of closely packed particles, measuring about 9 nm in diameter, and on the E face, the junctions were recognized as a shallow grooves.

• The Korean Journal of Zoology
• /
• v.26 no.4
• /
• pp.271-282
• /
• 1983

The authors observed the ultrastructure of the pigment cells of the frog, Rana nigromaculata Hallowell, during the hibernation. The specimens from the skin were fixed in 2.5% glutaraldehyde-paraform-aldehyde fixative in phosphate buffer at pH 7.2 prior to fixation in 2% osimium tetroxide, dehydrated in graded ethanol and acetone, embedded in Epon 812 mixture, and sectioned with LKB-ultramicrotome. the ultrathin sections were contrasted with uranyl acetate and lead citrate and observed with a JEOL-100B electron microscope. The results were as follows. In hibernating phase, pigment cells of the frog were consisted of the three kinds of chromatophores (xanthophore, iridophore and melanophore) in their dorsal skin. The traits of these cells were as follows. 1. Xanthophores A. Xanthophores were filled with pterinosomes and carotenoid vesicles. Many ribosomes, a few mitochondria and glycogen particles were dispersed in the cytoplasm. B. Pterinosomes were spherical or ellipsoidal in shape. They were divided into 6 types (type I, type II, type III, type IV, type V, type VI pterinosomes) by the their inner structure and especially, type I, type II, type III pterinosomes were well developed.

• Korean Journal of Veterinary Research
• /
• v.43 no.4
• /
• pp.541-549
• /
• 2003

The present study investigated the effects of aging on Leydig cells of Sprague Dawley rats. Rats of 3, 6, 12 and 18 months of age were used. Testes of rat were fixed by whole body perfusion using a fixative containing 2.5% glutaraldehyde in cacodylate buffer, processed and embedded in epon-araldite. Using $1{\mu}m$ sections stained with methylene blue, qualitative and quantitative morphological studies were performed. Testis incubations were used to determine luteinizing hormone (LH; 100 ng/ml) stimulated testosterone secretory capacity per testis in vitro. Testosterone levels in the incubation medium, and testosterone and luteinizing hormone levels in serum of these four groups of rats were determined by radioimmunoassay. Morphological studies revealed that Leydig cells were more abundant in the testis interstitium at 6, 12 and 18 months when compared with 3 months. The volumes of Leydig cells per testis was significantly higher, at 6, 12 and 18 months of age than those at 3 months. The number of Leydig cells per testis was doubled at 6, 12 and 18 months of age compared with 3 months. The average volume of a Leydig cell was not significantly different between 3 and 6 months of age, however, at 12 and 18 months a significantly lower value was observed. LH-stimulated testosterone production per testis in vitro was reduced by 45% at 6 months of age compared with 3 months; a further significant reduction was observed at 12 and 18 months. Serum testosterone and LH levels were not significantly different between 3 and 6 months of age but at 12 and 18 months a significantly lower value was observed in both groups for these hormones. These results showed that signs of aging are apparent in Leydig cells of Sprague Dawley rats at 12 months of age.

• Korean Journal of Ecology and Environment
• /
• v.37 no.4 s.109
• /
• pp.462-468
• /
• 2004

Most of the surveys of periphyton carried out for environmental and ecosystem health assessment of streams have considered the impact made on their fixative substrates (stones, rocks, sand, silt, clay and other abiotic matters), but there has been virtually no research that considered moving substrates. This study attempted to make an analysis and assessment of the habitat environments of the microalgae attached to the skin surfaces of fish living in small streams, with a focus on their species composition and community structure. The dominant fish in the this survey were Zacco temmincki and Zacco platypus, which are usually found in the streams, and rivers, and they accounted for 62% and 19%, respectively, in relative abundance. Substrates of fish, a representative organism with the trait of moving a long distance, show a marked contrast with those of organisms fixed at a certain place. Characteristics of both the upstream and the downstream reach well reflected in the microalgae attached to the skin surface of fishes, of which diatoms took the major composition. The result of this observation is considered to be useful to provide basic data in assessment of stream health. Also it may be suggested as a biological tool for the assessment of aquatic environment in the future.

• Journal of Periodontal and Implant Science
• /
• v.26 no.1
• /
• pp.27-46
• /
• 1996

The purpose of this study was designed to compare with the effects of 4 different surface active bioceramics on the healing process of alveolar bone defects in dogs. Artificial alveolar bone defects depth 4-6mm, width 3-4mm) were created with # 6 round bur at interproximal areas of maxillary canine, maxillary 2nd premolar, mandibular canine, and mandibular 3rd premolar. porous hydroxyapatite(Interpore $200^R$) , 45S5 bioglass, CJ4/lOC crystalline glass, and JJ crystalline glass were implanted in intrabony defects randomly. Experimental groups were divided into 4 categories according to its implant material. After implantation, all groups were examined postoperatively 4 weeks to 12 weeks. 3 dogs was selected randomly and sacrificed after vascular perfusion with 2.5% glutaraldehyde at every 4 weeks. Tissue blocks with surroundig alveolar bone and soft tissues were removed and immersed in formaldehyde/glutaraldehyde fixative. After 20 weeks decalcification with EDTA and formic acid, sections were made and observed under light microscope and transmission electron microscope. In all experimental groups, the encapsulation of inactive connective tissue was observed around graft particles in 4 weeks. As time elapsed, the thickness of surrounding connective tissue was decreased. Osteoconductive bone growth pattern was seen apparently in all groups. CJ4/lOC crystalline glass showed the most active bone formation until 8 weeks. 45S5 bioglass was, however, the most active in new bone formation at 12 weeks. Though there was difference in resorption rate among grafting materials, the size of graft particles was decreased gradually. 45S5 bioglass was resorbed faster than the others. On the other hand, porous hydroxyapatite was degraded most slowly. Phagocytosed particulate matters was observed in the cytoplasm of multinuclear multinuclear giant cell and macrophage under transmission electron microscope. The results suggested suggested that 45S5 bioglass and CJ4/lOC crystalline glass may have some enhanced reparative potential when compared to porous hydroxapatite in the treatment of periodontal defeds. JJ crystalline glass reguires a further investigation of the safety of its use.

• Asian-Australasian Journal of Animal Sciences
• /
• v.15 no.12
• /
• pp.1695-1701
• /
• 2002

The purposes of this study were to optimize the in vitro maturation (IVM) and culture (IVC) systems of rabbit oocytes. Cytoskeletal structures in the germinal vesicle stage (GV) and during IVM are also investigated. Ovaries were transported from local slaughterhouses and the cumulus-oocyte complexes (COCs) were collected from ovarian follicles (${\geq}1mm$). COCs were randomly allocated to TCM199-based medium ($T_1$, TCM-199) supplemented with $NaHCO_3$, glucose, sodium pyruvate and FSH ($T_2$), $T_2+E_2+LH$ ($T_3$), $T_3+FBS$ ($T_4$), or $T_1+E_2+LH+FSH+FBS$ ($T_5$), for IVM. In Experiment 1, COCs were retrieved from the follicles and 51 GV oocytes were fixed in the fixative (MTSB-XF) for nuclear and cytoplasmic examinations. In Experiment 2, progressive changes of both the nucleus and the cytoskeleton were examined at 0, 6, 16, and 20 h after IVM. Maturation (MR) and developmental rates were assessed in Experiment 3. Cytoplasmic microtubules (MT) were clearly observed in rabbit GV oocytes. To our knowledge, this is the first report that describes the appearance of MT structures in the GV stage ooplasm. Tremendous variations in cytoskeletal alterations were observed among treatments with the exception of the vitelline ring (VR), which is constantly visible and unchanged during maturation. Germinal vesicle breakdown (GVBD) does not occur at 6 h after onset of maturation culture. When the oocytes for IVM were collected within 2 h, results from Experiment 3 showed that rates of nuclear maturation were 42, 8, 42, 37 and 65% at 16 h of IVM for $T_1$ through $T_5$, respectively, in which $T_1$, $T_4$ and $T_5$ had significantly greater MR than those in other groups (p<0.05). Morula/blastocyst development after parthenogenetic activation ranged from 20 to 63% with significantly greater rates in $T_3$, $T_4$ and $T_5$ (p<0.05). These results suggested that oocytes recovered from slaughterhouse ovaries can be matured and parthenogenetically activated in vitro, but the MR remained low in this study. Addition of $E_2$ and LH in the medium may be beneficial for cytoplasmic maturation, but FBS exerts a nega- tive role in the subsequent development of parthenogenetic embryos when energy substrates are provided in the IVC media. More studies are required for improving the MR and further development of the GV stage rabbit oocytes.