• Maxillofacial Plastic and Reconstructive Surgery
• /
• v.27 no.3
• /
• pp.226-237
• /
• 2005

Purpose : Extensive defect of oral and maxillofacial area is usually reconstructed with composite flap including skin paddle. However, if the defects are lined with only skin components, the mucosa's role in mastication and texture are not restored. Furthermore, stiffness and hair-growing prevent denture rehabilitation and good oral hygiene. This study was performed to overcome the disadvantages of composite soft tissue flaps including the skin and to make a model for myo-mucosal flaps. Materials and methods : Buccal mucosa sized $0.5\times1.0\;cm^2$ from New Zealand rabbit (around 1.5kg) was harvested and cultivated by the modification of Rheinwald and Green's keratinocyte culture method. Cultured mucosa was grafted on the fascia of latismus dorsi as form of mucosal sheet. After 7, 10, 14 days, the myomucosal flap was excised and evaluated under light microscope with H & E and immunohistochemical staining. As control group, harvested buccal mucosa from rabbit was transplanted to gracilis muscle(n=6). Results : From 7 days after prelamination, the basal layer of the grafted mucosa resembled that of normal mucosa. As control group, transplanted mucosa had original shape but there's slight inflammatory reaction. Prelaminated mucosa has 19.8$\pm$4.59 cell layers and some samples have more than 20 layers. The expression rate of PCNA was relatively strong (42.9%$\pm$14.1) at the basal layer of grafted mucosa and the laminin was found at the basal layer. On the contrary, prelaminated mucosa at 10 days showed moderate expression rate of PCNA(32.4%$\pm$4.62). We found the mucosal layer was somehow disappeared and there is strong inflammatory reaction. After 14 days prelamination, the grafted oral keratinocytes were almost disappeared and expression of PCNA was not observed. Conclusion : We can make 75 fold large mucosal($3850mm^2$) sheet from small samples of mucosa $(50mm^2)$. Epithelial sheet that grafted on the fascia of muscle underwent differentiation and proliferation. But after 10, 14 days, there was strong inflammatory reaction and the grafted mucosa was destroyed from surface layer. In rabbit model, transfer of fascio-mucosal flap should be done from 7 to 10 days after prelamination.

• Journal of Periodontal and Implant Science
• /
• v.23 no.1
• /
• pp.77-96
• /
• 1993

This in vitro study was undertaken to observe whether citric acid application aids the attachment and proliferation of human periodontal ligament cells to the root surfaces of periodontally diseased teeth. The roots were prepared so that the comparison could be made among the control healthy root surface, citric acid demineralized and non-demineralized root planted surfaces. Prior to the cell attachment experiment, each groups were prepared for scanning electron microscopic (SEM) examinations of root surface morphology, All specimens were fixed with phosphate buffered glutaraldehydes, postfixed with phosphate buffered osmium tetraoxide and stained with phosphate buffered tannic acid. dehydrated in ethanol, critical point dried, sputter coated with gold and examined under the SEM. In the cell attachement experiment, human cultured periodontal ligament cells at concentration to $4.5{\times}\;10^4\;cells/ml$ were seeded in each culture well which contained prepared roots and incubated for 30min 1, 2, 6, 12 and 24 hours at 37, 5% $CO_2$air incubator. Than the specimens were prepared for SEM examination using, the same methods as described above. In the cell proliferation experiment, $5{\times}\;10^4\;cells/ml$ cells were seeded incubated with the specimens for 6 hours. Then, all of the specimens were moved into fresh culture well and incubated for 24, 48, and 72 hours. The cell counting was done after trypsinization, under light microscope. The results were as follows. When viewed the surface morphology prior to the cell attachment, the non acid treated root planed surface displayed scaling striation and occasional bacteria and calculus. The citric acid treated specimens displayed little debris on the surface and funnel shaped orifices of dentinal tubules. There were no apparent differences in the morphology of cells attached to the control and experiment groups. However, in initial attachement, there was a slight more enhanced appearance in attachment in citric acid treated groups than other root surfaces. After 6 hours of incubation, most of the cells initiated the alteration of cell morphology from ovoid to spindle shapes. After 24 hours of incubation, most of the cells displayed proliferated appearance and connected with each other via numerous processes. In the cell proliferation experiments, there were statistically significant increased number of cells in citic acid treated groups than other groups.

• Journal of Radiation Protection and Research
• /
• v.37 no.4
• /
• pp.208-212
• /
• 2012

In previous studies, ovarian follicle in rat has been used a higher radiation dose than that for cancer radiotherapy in clinical practice. The aim of this study was to evaluate the effect of radiation dose used for cancer radiotherapy on ovarian follicle atresia in rat. Mice of 4-week-old female were whole body irradiated with 2 cGy or 2 Gy (Mevatron 67, Siemens, Germany) and sacrificed by cervical dislocation. Ovaries were collected at 24 hours after irradiation to observe the degree of follicular atresia. Ovaries were fixed in neutral formaldehyde solution for 24 hours and embedded with paraffin. Cutted in $5{\mu}m$ thickness with microtome and stained with hematoxylin and eosin (H&E) and TUNEL immunohistochemical stain, and examined histologically under a light microscope. All data were presented as mean ${\pm}SD$, calculating the ratio of normal or atretic follicles to total ovarian follicles. Statistical analysis was performed by the Mann Whitney test using the SPSS ver 19.0. Ratio of atretic to total follicles of 2 Gy group was significantly higher than control or 2 cGy groups (p<0.05). Ratio of normal to total follicles of 2 Gy group was significantly lower than control group in preantral follicle (64.0 vs. 87.7, p=0.027). Ratio of normal to total follicles of 2 cGy group was significantly increased more than control or 2 Gy groups in antral follicle, and there were no significant difference between control and 2 Gy groups (p=0.522). Radiation dose of 2 Gy for cancer radiotherapy have a significant effect on ovarian follicle atresia in rat.

• Biomedical Science Letters
• /
• v.4 no.1
• /
• pp.65-72
• /
• 1998

Recently there have been frequent reports on human infection caused by the Diphyllobothridae in Korea. The adequate opportunities for Koreans to eat raw fish, the primary infection medium of cestodes and the human infection through drinking water by cyclops, the first intermediate host are believed to be main reasons for the infection. The first task of this study was to classify and diagnose the species by differentiating morphological characteristic between scolex and proglottids of cestodes. However, the initially available diagnosis was done with the patient's symptoms and the eggs obtained from his stool. It is important to differentiate the species by the eggs of Diphyllobothrium latum especially in that it can help get advance information for a more reliable analysis in the near future. The morphological and diagnostic results from proglottids and eggs of Diphyllobothrium latum, Diphyllobothrium latum parvum and Spirometra erinacei are as follows； In each kind of cestodes from the patient's stool, the shape and size of 50 eggs were measured. Eggs of Diphyllobothrium latum had an operculum and were ovoidal or ellipsoid to elliptical in shape. Eggs of Diphyllobothrium latum parvum were more ovoidal in shape and smaller in size than Diphyllobothrium latum. And eggs of Spirometra erinacei were asymmetrical in width and long and slender in shape. The average lengths and widths of Diphyllobothrium latum, Diphyllobothrium latum parvum and Spirometra erinacei were 61.4$\times$41.7 $\mu\textrm{m}$, 55.9$\times$41.4 $\mu\textrm{m}$ and 66.7$\times$36.4 $\mu\textrm{m}$, respectively. After the segments of each cestode were fixed, embedding and hematoxylin-eosin dyeing on a microtome-made specimen were done. The micrographs of the semicon's aceto-carmine dyed specimen showed that Diphyllobothrium latum and Diphyllobothrium latum parvum had a centrally-located genital gland and an opened uterine pore. The yolks were observed on both sides of proglottids and had a typical rosette pattern. Yet, Diphyllobothrium latum parvum was shown smaller than Diphyllobothrium latum in the micrograph. Proglottids of Spirometra erinacei displayed that the uterus was rolled spirally more than five to seven times, and connected successively to the seminal vesicle in the cirrus sac. Shown above, this study was performed to measure the size of eggs and analyze the morphological characteristics of proglottids and provided the measurements of three types of cestodes obtained by a light microscope.

• Korean Journal of Plant Tissue Culture
• /
• v.24 no.1
• /
• pp.1-35
• /
• 1997

Fertilized egg, by successive cell divisions, differentiates into different tissues and organs with various structures and functions. Different cells and tissues contain different proteins, products of selective gene expression. Not all the genes in any genomes are equally active, temporal and spatial gene expression being the general rule. Present paper attempts to review the tanscriptional mechanisms or the initiations of transcription from several angles. In some of the organisms the genes in the process of transcription or the genes in the inactive state can be seen under the light microscope. Some bands of Drosophila polytene chromosomes may exhibit a swollen or puff appearance under certain conditions. A puff, unfolded or decondensed form of chromomere, represents sets of intense transcriptional activity or RNA synthesis. The heterochromatic X chromosome whose genes remain inactive in the female mammals can be visualized as a dark staining structure called Barr body, Configuration of chromatin differs between transcribed and nontranscribed chromatin. Modification to the chromatin facilitates RNA synthesis. The movement of large polymerase molecule along the DNA would probably be facilitated if some modifications of the chromatin configuration is effected. Methylation of cytosines in CG sequences is associated with inactive genes. Methylation can play a role in determination of mammalian cells during embryogenesis. Demethylation is necessary for the gene to be expressed during development A histone modification that is also known to be correlated with transcriptional capacity of chromatin is acetylation of the lysine residues of the core histones. Chromatin containing a high level of histone acetylation is very sensitive to DNase 1. For the transcription to occur TBP must first bind to the TATA box. Another TF, TF IIB, then binds to the promoter-TBP complex, facilitating the access of RNA polymerase to the transcription initiation site. As recently as eight years ago researchers assumed that histones were irrelevant to the regulation of gene expression. Histones combine with the DNA to form nucleosome of the chromatin. Histones are vital participant in gene regulation. Histone and basal factors compete for access to TATA box. When DNA is exposed to basal factors before histones are introduced, the basal factors assemble on TATA boxes preventing the access of histones, allowing transcription to occur, for transcription to begin, activator protein at the upstream activation sequence or enhancer must interact with the tail of histone H4 at TATA box and cause the histone role particle to dissociate from the TATA box leading to partial breakup of the histone core particle and allowing the basal factors to bind to the TATA box. New concept of genomic flux in contrast to the old concept of static genome has been developed based on the powerful new molecular techniques. Genomic changes such as repetitive DNAs and transposable elements, it is assumed but not yet proved, may affect some of the developmental patterns that characterize particular cells, tissues, organs, and organisms. In the last decade or so remarkable achievement have been made in the researches of the structures and functions of TFs and the specific target sequences located in promoters or enhancers where these TFs bind. TFs have independent domains that bind DNA and that activate transcription. DNA binding domain of TFs serves to bring the protein into the right location. There are many types of DNA binding domains. Common types of motifs can be found that are responsible for binding to DNA. The motifs are usually quite short and comprise only a small part of the protein structure. Steroid receptors have domains for hormone binding, DNA binding, and activating transcription. The zinc finger motif comprises a DNA binding domain. Leucine zipper consist of a stretch of amino acids with a leucine residue in every seventh position Two proteins form a dimer because they interact by means of leucine zippers on similar α-helical domain. This positions their DNA binding basic domains for interaction with the two halves of a DNA sequence with dyad symmetry of TGACTCA, ACTGAGT.

• Journal of Korean Ophthalmic Optics Society
• /
• v.9 no.2
• /
• pp.333-343
• /
• 2004

Impression cytology refers to application of cellulose acetate filter material to the ocular surface to remove the superficial layers of the conjunctival epithelium. The technique is non-invasive, is easy to perform, causes minimal discomfort to the patient, and can be used to follow changes in the conjunctival ocular surface over time. With this method, the morphology of the conjunctival ocular surface can be studied and the degree of squmaous metaplasia assessed. This study was performed to evaluate the conjunctival surface by impression cytology in dry eye patients. A total of 70 students with no contact lens wearing history were recruited. Subjects were required to fill in a McMonnies dry eye symptom questionnaire. The non-invasive tear thinning time(TIT) test of each subject was measured, followed by Schirmer tear test(STI), tear film break-up time(TBUT) tests and Rose-bengal staining were performed as a baseline. Conjunctival epithelial cells from the inferior bulbar conjunctiva were harvested by the impression cytology technique. The specimens collected were labelled and stained with PAS(Periodic Acid Schift)-haematoxylin. The goblet cells and conjunctival epithelial cells were observed under a light microscope of 400x magnification. The specimens were classified according to the Nelson Grading scale which was based on the degree of squamous metaplasia such as changes of goblet cells density, size/form, N:C(nucleus : cytoplasm) ratio. Dry eye patients were observed morphological changes of the epithelial cells, different nuclear alterations, decrease of the goblet cells density. The degree of cytological changes was related to severity of dry eye conditions. When the epithelial cell morphology was graded according to the system described by Nelson, specimens from the control group revealed 91.43% of the eyes to be grade 0 and 8.57% to be grade 1, whereas of the dry eye patients, 20% were grade 0, 42.86% grade 1, 34.29% grade 2 and 2,86% grade 3. Impression cytology represents a non- or minimally invasive biopsy of the ocular surface epithelium with no side effects or contraindications. It has demonstrated to be a useful diagnostic aid for a wide variety of processes involving the ocular surface. This technique is a safe, simple method and may help increase understanding of various ocular surface alterations in dry eye patients.

• Journal of Chest Surgery
• /
• v.42 no.1
• /
• pp.1-8
• /
• 2009

Background: The pathophysiology of acute respiratory distress syndrome with sepsis is acute lung injury (ALI) that's' caused by endotoxin (LPS). We evaluate effects of moxifloxacin on LPS-induced ALI in a rat model. Material and Method: The rats were divided into 3 groups as the control group (C), the LPS insult group (L), and the LPS+moxifloxacin treated group (L-M). ALI was induced by endotracheal instillation of E.coli LPS, then moxifloxacin was given in 30 minutes. Five hours later, we checked the lung weight/body weight ratio(the L/BW ratio), the protein & neutrophils in the bronchoalveolar lavage fluid (BALF), the myeloperoxidase (MPO) activity & the malondialdehyde (MDA) content, the expressions of cytosolic and secretory phospholipase $A_2$ (c, $sPLA_2$), and the morphology of the lung with using a light microscope. Result: The L/BW ratio, the protein content and the neutrophil count in the BALF, and the MPO activity and the MDA content in lung were significantly increased in group L compared to group C, and these factors were markedly decreased in group L-M compare to group L. The $cPLA_2$ expression and the $sPLA_2$ expression were increased in group L and the $cPLA_2$ expression was decreased in group L-M. Yet the $sPLA_2$ expression was not changed in group L-M. Morphologically, many inflammatory findings were observed in group L, but not in group L-M. Conclusion: Many of the inflammatory changes of ALI that were caused by LPS insult were ameliorated by moxifloxacin treatment.

• Applied Microscopy
• /
• v.11 no.1
• /
• pp.59-65
• /
• 1981

The pars distalis of the Korean frogs (Rana dybowskii Guenther) during hibernating and active periods was observed with the electron microscope. Seven cell types were classified according to the size and shape of secretory granules and to the ultrastructural characteristics. There were many differences between hibernating and active frogs in type 5 cells. Therefore the following results were observed. Cell type 1; This type cell contains spherical secretory granules, $375{\sim}687m{\mu}$ in diameter. Cell type 2; This type cell contains various secretory granules, $250{\sim}437m{\mu}$ in diameter Cell type 3; Spherical and rod-shaped granules, $l25{\sim}187m{\mu}$ in diameter were observed. Cell type 4; In this type cell, the electron density is the lowest and the density of granules is the highest of all type cells. This type cell contains various secretory granules and large secretory granules, $2l0{\sim}420m{\mu}$ in diameter, were also observed. Cell type 5; The electron density of this cells is similar to that of type 4 cells. The density of granules is lower than that of type 4 cells. And the shapes of the secretory granules are similar to those of type 4 cells. But many rod shaped granules, $200{\sim}863m{\mu}$ in diameter, were also observed. Cell type 6; This type was similar to type 2. The electron density of cytoplasm is very low. Spherical granules, $232{\sim}316m{\mu}$ in diameter, were observed. Cell type 7; This type of cell has no secretory granules. This cell is not developed very well. The type 5 cells in hibernating frogs are different from cells in active frogs. In type 5 cells, many secretory granules were observed during active period. But the number of secretory granules were greatly declined and there were many vacuoles in cytoplasm during hibernating period.

• The Korean Journal of Malacology
• /
• v.27 no.4
• /
• pp.359-369
• /
• 2011

Twelve species of food microalgae were investigated to clarify the digestion index of Crassostrea gigas larvae using epifluorescence microscopy to choose an appropriate diet for artificial seed production in hatchery. An experiment was conducted using 1 (D shaped stage), 4 (Early umbo stage), 8 (umbo stage) and 12 (Full grown stage) days old larvae. larvae were stocked in 1 L flasks at 5 individuals/mL and fed $10{\times}10^4$ algal cells/mL of each species individually. Prior to larvae were fed for 3 h and then were observed under the microscope to detect ingestion; larvae were then sieved and replaced in 1 L flasks containing filtered seawater and were observed after 3, 5 and 8 h to analyse the digestion index. Values of digestion indices were specific for each alga. No evidence for the ingestion of Thalassiosira weissflogii was evident at all larval development stages tested. Digestion indices of others microalgae were 0.8-99.7% at 4 stage of larval development stages: Chlorella ellipsoidea (0.8-5.4%), Nannochloris oculata (1.4-5.0%), Isochrysis galbana (99.1-99.5%), Pavlova lutheri (99.1-99.5%), I. aff. galbana (99.4-99.5%), Cheatoceros calcitrans (0.0-99.2%), C. gracilis (0.0-99.7%), C. simplex (0.0-95.9%), Phaeodactylum tricornutum (0.0-99.6%), Tetraselmis tetrathele (0.0-99.7%) and Dunaliella tertiolecta (0.0-99.6%), respectively. Therefore, it is assumed that food microalgae showing the high digestion such as I. galbana should be supplied to the early umbo stage larvae, and then after the umbo larval stage, the mixed microalgae with diatoms and light green algae should be supplied to the full grown stage larvae to increase the digestion of their larvae.

• Korean Journal of Veterinary Research
• /
• v.43 no.1
• /
• pp.11-24
• /
• 2003

This experimental studies was to investigate the location of PNS and CNS labeled neurons following injection of 2% WGA-HRP and pseudorabies virus (PRY), Bartha strain, into the ductus deferens of rats. After survival times 4-5 days following injection of 2% WGA-HRP and PRV, the rats were perfused, and their brain, spinal cord, sympathetic ganglia and spinal ganglia were frozen sectioned ($30{\mu}m$). These sections were stained by HRP histochemical and PRY inummohistochemical staining methods, and observed with light microscope. The results were as follows ; 1. The location of sympathetic ganglia projecting to the ductus deferens were observed in pelvic ganglion, inferior mesenteric ganglion and L1-6 lwnbar sympathetic ganglia. 2. The location of spinal ganglia projecting to the ductus deferens were observed in T13-L6 spinal ganglia. 3. The PRY labeled neurons projecting to the ductus deferens were observed in lateral spinal nucleus, lamina I, II and X of cervical segments. In thoracic segments, PRY labeled neurons were observed in dorsomedial part of lamina I, II and III, and dorsolateral part of lamina IV and V. Densely labeled neurons were observed in intermediolateral nucleus. In first lumbar segment, labeled neurons were observed in intermediolateral nucleus and dorsal commisural nucleus. In sixth lumbar segment and sacral segments, dense labeled neurons were observed in sacral parasympathetic nuc., lamina IX and X. 4. In the medulla oblongata, PRV labeled neurons projecting to the ductus deferens were observed in the trigeminal spinal nuc., A1 noradrenalin cells/C1 adrenalin cells/caudoventrolateral reticular nuc., rostroventrolateral reticular nuc., area postrema, nuc. tractus solitarius, raphe obscurus nuc., raphe pallidus nuc., raphe magnus nuc., parapyramidal nuc., lateral reticular nuc., gigantocellular reticular nuc.. 5. In the pons, PRV labeled neurons projecting to the ductus deferens were ohserved in parabrachial nuc., Kolliker-Fuse nuc., locus cooruleus, subcooruleus nuc. and AS noradrenalin cells. 6. In midbrain, PRV labeled neurons projecting to the ductus deferens were observed in periaqueductal gray substance, substantia nigra and dorsal raphe nuc.. 7. In the diencephalon, PRV labeled neurons projecting to the ductus deferens were observed in paraventricular hypahalamic nuc., lateral hypothalamic nuc., retrochiasmatic nuc. and ventromedial hypothalamic nuc.. 8. In cerebrum, PRV labeled neurons projecting to the ductus deferens were observed in area 1 of parietal cortex. These results suggest that WGA-HRP labeled neurons of the spinal cord projecting to the rat ductus deferens might be the first-order neurons related to the viscero-somatic sensory and sympathetic postganglionic neurons, and PRV labeled neurons of the brain and spinal cord may be the second and third-order neurons response to the movement of smooth muscles in ductus deferens. These PRV labeled neurons may be central autonomic center related to the integration and modulation of reflex control linked to the sensory and motor system monitaing the internal environment. These observations provide evidence for previously unknown projections from ductus deferens to spinal cord and brain which may be play an important neuroanatornical basic evidence in the regulation of ductus deferens function.