Expression of enamel proteins in rough endoplasmic reticulum and golgi complex in human dental germs / Expresión de proteínas del esmalte en retículo endoplásmico rugoso y complexo golgiensis en gérmenes dentales humanos

Tooth enamel is the hardest tissue in the body. The organic matrix configuration is provided by the main proteins amelogenin, ameloblastin and enamelysin (MMP20), an enzyme that helps to shape the matrix. The aim of this study was to determine by histochemistry the expression of amelogenin and enamelysin through the rough endoplasmic reticulum in the late stages of amelogenesis, and its expression in the Complexus golgiensis (Golgi complex / Golgi apparatus) in the early stages in human fetuses. In early stages a colocalization of both proteins inside the Golgi apparatus was found, being more evident the relationship between Golgi and amelogenin (99.92 %). In the late stage, a colocalization of both proteins and rugged endoplasmic reticulum was found. With enamelysin being more evident in relation with rough endoplasmic reticulum (99.95 %). Our findings demonstrated the presence of amelogenin and enamelysin in odontoblast and ameloblast. However, the presence of these two proteins in odontoblast remains unknown.

El esmalte dental es el tejido más duro del cuerpo. La configuración de la matriz orgánica es proporcionada por las proteínas principales amelogenina, ameloblastina y enamelisina (MMP20), una enzima que ayuda a dar forma a la matriz. El objetivo de este estudio fue determinar mediante histoquímica la expresión de amelogenina y enamelisina a través del retículo endoplasmático rugoso en las últimas etapas de la amelogénesis , y su expresión en el Complexo golgiensis en las primeras etapas de formación en fetos humanos. En las primeras etapas se observó colocalización de ambas proteínas en el interior del Complexo golgiensis, siendo más evidente la relación entre Golgi y amelogenina (99,92 %). En la última etapa, se identificó una colocalización de ambas proteínas y retículo endoplásmico rugoso. Resulto más evidente la enamelisina en relación con el retículo endoplasmático rugoso (99,95 %). Nuestros resultados demostraron la presencia de amelogenina y enamelisina en odontoblastos y ameloblastos, sin embargo se desconoce la presencia de estas dos proteínas en odontoblastos.

5-Aza-2'-deoxycytidine acts as a modulator of chondrocyte hypertrophy and maturation in chick caudal region chondrocytes in culture / 대한해부학회지

This study was carried out to explore the effect of DNA hypomethylation on chondrocytes phenotype, in particular the effect on chondrocyte hypertrophy, maturation, and apoptosis. Chondrocytes derived from caudal region of day 17 embryonic chick sterna were pretreated with hypomethylating drug 5-aza-2'-deoxycytidine for 48 hours and then maintained in the normal culture medium for up to 14 days. Histological studies showed distinct morphological changes occurred in the pretreated cultures when compared to the control cultures. The pretreated chondrocytes after 7 days in culture became bigger in size and acquired more flattened fibroblastic phenotype as well as a loss of cartilage specific extracellular matrix. Scanning electron microscopy at day 7 showed chondrocytes to have increased in cell volume and at day 14 in culture the extracellular matrix of the pretreated cultures showed regular fibrillar structure heavily embedded with matrix vesicles, which is the characteristic feature of chondrocyte hypertrophy. Transmission electron microscopic studies indicated the terminal fate of the hypertrophic cells in culture. The pretreated chondrocytes grown for 14 days in culture showed two types of cells: dark cells which had condense chromatin in dark patches and dark cytoplasm. The other light chondrocytes appeared to be heavily loaded with endoplasmic reticulum indicative of very active protein and secretory activity; their cytoplasm had large vacuoles and disintegrating cytoplasm. The biosynthetic profile showed that the pretreated cultures were actively synthesizing and secreting type X collagen and alkaline phosphatase as a major biosynthetic product.

Efecto de estimulaciones infrarrojas sobre hepatoesteatosis microvesicular alcohólica de ratas / Infrared laser effect on alcoholic rat hepatic microvesicular steatosis

24 ratas hembras de 4 meses de vida con peso aproximado de 250 gramos fueron divididas en dos grupos de animales, A y B. Ambos grupos se mantuvieron con pellet y solución de alcohol 40% durante 60 días generándoseles una hepatoesteatosis microvesicular. Los hígados de los animales pertenecientes al grupo B fueron estimulados con láser infrarrojo 6 J/cm2 durante 15 días consecutivos. Posteriormente, las ratas fueron sacrificadas y se extrajeron muestras de hígado y luego procesadas para microscopía electrónica de transmisión. De ambos tipos celulares se obtuvieron microfotografías electrónicas de transmisión con aumentos finales de 8.500 X, las cuales fueron sometidas a estudios morfométricos para determinar fracciones volumétricas de los siguientes componentes celulares: Retículo endoplasmático rugoso (RER), mitocondrias, inclusiones lipídicas y de glicógeno, eu y heterocromatina. De igual manera se cuantificaron las áreas celulares y nucleares. Del análisis de los resultados entre hepatocitos esteatósicos e irradiados se visualiza que existen diferencias en todos los componentes celulares cuantificados y se concluye que los efectos de la estimulación infrarroja con dosis de 6 J/cm2 provoca en los hepatocitos con esteatosis microvesicular transformación en su ultraestructura y en su morfología, fundamentalmente en la disminución acentuada de las infiltraciones lipídicas hasta en un 80% situación que se traduciría, en una variación funcional, representando de esta manera un efecto evidente que estas inducciones infrarrojas generan.

24 four-month-old female rats weighing approximately 250 grams were divided into two groups labeled A and B. Both groups were fed pellets and a 40% alcohol solution for 60 days, which caused a microvesicular hepatic steatosis. The livers of the animals in Group B were stimulated with 6 J/cm2 of infrared laser for 15 consecutive days. The rats were then sacrificed and samples of both steatosic liver and liver stimulated with infrared inductions were extracted for immediate processing via transmission electron microscopy.From both cell types transmission electron microphotographs were obtained at magnifications of 9500 X; these were subjected to morphometric studies to determine volumetric fractions of the following cell components: rough endoplasmic reticulum (RER), mitochondria, lipid and glycogen inclusions, euchromatin and heterochromatin. Likewise, cell and nuclear areas were quantified. Analysis of the results between steatosic and radiated hepatocytes revealed notable differences in all the cell components quantified. It is concluded that the effects of infrared stimulation with a dose of 6 J/cm2 brings about in the steatosic hepatocytes a microvesicular transformation in their ultrastructure and morphology, fundamentally in the considerable decrease in lipid infiltrations to 80%, which ultimately translates into a functional variation, thus representing an obvious impact produced by these infrared inductions.

Transient Bullous Dermolysis of the Newborn / 대한피부과학회지

Transient bullous dermolysis of the newborn (TBDN) is a rare subtype of the dystrophic epidermolysis bullosa characterized by blistering at birth which improves spontaneously during early life. Electron microscopy showed sublamina densa separation with dilated rough endoplasmic reticulum and electron dense inclusions. Immunofluorescence mapping using anti-type VII collagen antibody showed widespread intraepidermal type VII collagens which are a characteristic finding of TBDN. Here, we report two cases of TBDN presenting typical clinical manifestations, electron microscopy findings, and immunofluorescence mapping results. The skin lesions of both patients healed spontaneously 2~3 months later.

A Case of Unilateral Electric Cataract with Optic Neuropathy after Electric Injury

PURPOSE: Electric injury may result in cataracts, which usually occur bilaterally. In this report, we present a rare complication of such an injury presenting as a unilateral cataract with optic neuropathy. CASE SUMMARY: A 17-year-old male patient presented with gradual worsening of vision in his right eye 7 weeks after sustaining an injury from 22,900 volts of high-voltage electric current his right hand. On initial eye examination, the best corrected visual acuity (BCVA) was 20/60 in the right eye and 20/20 in the left eye. Relative afferent pupillary defect (RAPD) was noted in the right eye. Slit-lamp examination confirmed the typical anterior subcapsular lenticular opacities and funduscopy showed a slightly pale optic nerve head in the right eye. Pattern reversal visually evoked potential (P-VEP) showed a significant delay of P-100 implicit time in the right eye. After 7 months, phacoaspiration and posterior chamber intraocular lens implantation were performed in the right eye. Postoperatively, the BCVA improved to 20/30 but visual field examination showed a partial defect in the temporal area of the right eye. Cataract did not develop in the left eye during 15 months of follow-up. The electron microscopic findings showed that the number of mitochondria in the cytoplasm increased. The rough endoplasmic reticulum in the cytoplasm and microfilaments were enriched. CONCLUSIONS: Manifestation time of ocular complications after electric injuries is different. Therefore an ophthalmic examination should be performed regularly in the early recovery period of such injuries and in particular, progression of bilateral electric cataract must be checked.

Solitary Fibrous Tumor of the Conjunctiva with Heretofore Undescribed Pathologic Findings

A 37-year-old female presented with a conjunctival mass discovered 3 years prior. An excisional biopsy revealed a patternless proliferation of round and spindle-shaped cells with an eosinophilic fibrillary cytoplasm and vesicular nuclei with occasional inclusions. Psammoma bodies were arranged around the dilated irregularly-shaped vessels. Differential diagnoses included conjunctival solitary fibrous tumor (SFT), nevus, glomangioma, ectopic meningioma, and mesectodermal leiomyoma. The tumor cells were immunoreactive for CD34, CD99, bcl-2 and vimentin, and were negative for smooth muscle actin, desmin, glial fibrillary acidic protein, S-100 protein, epithelial membrane antigen, and human melanoma black-45. Ultrastructurally, the tumor cells had rough endoplasmic reticulum, free ribosomes, and scattered mitochondria without basal lamina or cellular junctions, which are features of fibroblasts. A diagnosis of SFT was rendered based on the light microscopic, immunohistochemical, and electron microscopic findings. We report here on the second case of a SFT arising in the conjunctiva, which clinically and histologically mimics conjunctival nevus, glomangioma, ectopic meningioma, and a hybrid neurogenic-myogenic tumor such as mesectodermal leiomyoma.

Ultrastructural changes of the olfactory bulb in manganese-treated mice

The effect of manganese toxicity on the ultrastructure of the olfactory bulb was evaluated. Male albino mice were injected intraperitoneally with MnCl2 (5 mg/Kg/day) five days per week during nine weeks. The control group received NaCl (0.9%). The olfactory bulbs of five mice from each group were processed for transmission electron microscopy after 2, 4, 6 and 9 weeks of manganese treatment. On week 2, some disorganization of the myelin sheaths was observed. After 4 weeks, degenerated neurons with dilated cisternae of rough endoplasmic reticulum and swollen mitochondria appeared. A certain degree of gliosis with a predominance of astrocytes with swollen mitochondria, disorganization of the endomembrane system, dilation of the perinuclear cisternae and irregularly shaped nuclei with abnormal chromatin distribution were observed after 6 weeks. Some glial cells showed disorganization of the Golgi apparatus. On week 9, an increase in the number of astrocytes, whose mitochondrial cristae were partially or totally erased, and a dilation of the rough endoplasmic reticulum were found. Neurons appear degenerated, with swollen mitochondria and a vacuolated, electron dense cytoplasm. These changes seem to indicate that the olfactory bulb is sensitive to the toxic effects of manganese.

Ultrastructural features of Mimulus aurantiacus (Scrophulariaceae) pollen tubes in vivo

The aim of this study is to give information on ultrastructure of in vivo pollen tubes of Mimulus aurantiacus which were collected from the Botanical Garden of the University of California at Berkeley. Materials were prepared according to electron microscopy methods and examined under Zeiss electron microscope. Four zones were examined in the pollen tubes of Mimulus aurantiacus. APICAL ZONE: Mitochondria, smooth endoplasmic reticulum, rough endoplasmic reticulum, dictyosomes and secretory vesicles were observed. SUBAPICAL ZONE: This area contained abundant rough endoplasmic reticulum and occasionally some smooth endoplasmic reticulum. The polysomes, mitochondria, proplastids that contain starch, small vacuoles and a few lipid bodies were detected. NUCLEAR ZONE: Both generative and vegetative cell nuclei lie in this zone. The vegetative cell nucleus was large and long. Rough endoplasmic reticulum, mitochondria, ribosomes, dictyosomes, and amyloplasts that are rich of starch were observed. VACUOLATION AND PLUG FORMATION ZONE: Cytoplasm of the tubes was full of large vacuoles. Few organelles such as mitochondria, dictyosome and rough endoplasmic reticulum were detected along their periphery.

O objetivo deste estudo é informar sobre a ultraestrutura de tubos de pólen de Mimulus aurantiacus in vivo coletados no "Botanical Garden" da Universidade da Califórnia em Berkeley. O material foi preparado de acordo com os métodos de microscopia eletrônica e examinado em microscópio eletrônico Zeiss. Quatro zonas dos tubos de pólen de Mimulus aurantiacus foram examinadas. ZONA APICAL: foram observados mitocôndrias, retículo endoplasmático liso; retículo endoplasmático rugoso, dictiossomos e vesículas secretoras. ZONA SUBAPICAL: esta área continha retículo endoplasmático rugoso em abundância e, ocasionalmente, algum retículo endoplasmático liso. Foram detectados polissomos, mitocôndrias, proplastídeos que contêm amido, pequenos vacúolos e alguns corpos lipídicos. ZONA NUCLEAR: nesta área, existem tanto núcleos de células geradoras como vegetativas. O núcleo de célula vegetativa é grande e longo. Foram observados retículo endoplasmático rugoso, mitocôndria, ribossomos, dictiossomos e amiloplastos ricos em amido. ZONA DE VACUOLIZAÇÃO E DE FORMAÇÃO DE "PLUG": o citoplasma dos tubos estava cheio de grandes vacúolos. Algumas organelas como mitocôndria, dictiossomo e retículo endoplasmático rugoso foram detectadas em toda a periferia desta área.

Histological, scanning and transmission electron microscopic studies on the possible protective role of ginger extract against acrylamide induced intestinal damage in mice

This study was carried out to evaluate the protective effect of ginger Zingeber officinale extract [ZOE] against the acrylamide [AC] which is an industrial chemical used in water treatment and it is synthesized during cooking of starch food at high temperature. Thirty adult male albino mice, each weighs 20-25 g were divided into three groups [10 mice/group]: [I]control group. II]acrylamide treated group. [III] acrylamide and ginger group. Acrylamide was given to experimental animals in the drinking water at a non-lethal dose of 200 p.p.m for 10 weeks [3 days/week]. Ginger extract was orally administrated at 50 mg/L [tilde 5 ml/day] for 10 weeks [3 days/week]. The ileum samples were collected for light microscope study and for scanning and transmission electron microscope examination. This study revealed that acrylamide induces pathological changes of the ileum of the treated mice specially the absorptive epithelial cells. The scanning electron microscopic study revealed damage of the ileal villi, some red blood corpuscles appeared at the site of damage. The transmission electron microscopic examination clearly demonstrated degeneration of most cell organelles as mitochondria, deterioration and degranulation of the rough endoplasmic reticulum, dilatation of Golgi apparatus. The administration of ginger extract decreased the histological alterations and ensuring the anti-inflammatory, and antitoxic effects of ZOE at its chosen dosage level

Tubeimoside I-induced ultrastructural changes of human cervical carcinoma HeLa cell line and the protective effect of cyclosporine A / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the ultrastructural changes of HeLa cells in response to tubeimoside I (TBMS1) treatment and the protective effect of cyclosporine A (CsA), and explore the role of intrinsic apoptosis pathway in TBMS1-induced HeLa cell apoptosis.</p><p><b>METHODS</b>HeLa cells were treated with TBMS1 (10-50 micromo/L) alone or in combination with 2 micromol/L CsA for 12 and 24 h and observed with transmission electron microscope (TEM) for the ultrastructural changes of the cells.</p><p><b>RESULTS</b>TBMS1 induced apoptosis of HeLa cells in a concentration- and time-dependant manner. Under TEM, the treated cells progressively shrunk and the intercellular space widened with loss of microvillus, mitochondrial swelling, rough endoplasmic reticulum enlargement, chromatin condensation, nuclear shrinkage and nuclear pyknosis as TBMS1 concentration increased. At low concentrations, CsA offered partial protection of the mitochondria from TBMS1-induced damage whereas high-concentration CsA did not.</p><p><b>CONCLUSION</b>TBMS1 induces ultrastructural changes typical for apoptosis of the HeLa cells, which provides morphological evidence for the role of intrinsic apoptosis pathway in TBMS1-induced apoptosis.</p>

Fine Structure of Goblet Cell Regeneration on Experimental Colitis Induced by Dextran Sulfate Sodium / 대한해부학회지

Ulcerative colitis is recognized as important causes of gastrointestinal diseases in children and adults. I observed the fine structural changes of goblet cell regeneration after experimental ulcerative colitis induced by dextran sulfate sodium (DSS) in rats. Experimental colitis was induced with 5% DSS in the drinking water for 5 days, and the healing groups were fed with pure water for 7 days thereafter. In the early stage of goblet cell regeneration (repair 3 days group), granular endoplasmic reticulums were developed around the nucleus, and some mucigen granules were observed around the nucleus. In the middle stage of goblet cell regeneration (repair 5 days group), Golgi complexes were well developed in the upper region to the nucleus, and many mucous granules were observed. In the matured goblet cell regeneration (repair 7 days group), many mucous granules appeared in the upper region of the cell, and cell organelles were located in the base and periphery of the cell. These results suggest that the goblet cell was completely reconstructed within 7 days after ulcerative colitis.

Glycosaminoglycan expression in pleomorphic adenomas of the salivary gland

Ubisch bodies and pollen ontogeny in Oxalis articulata Savigny

The correlation between the ontogeny of Ubisch bodies and pollen development in Oxalis articulata was studied with Transmission Electron Microscopy (TEM). The ultrastructural changes observed during the different stages of development in the tapetal cells are related to Ubisch bodies, sporopollenin and pollen-kitt formation. The pro-orbicules have the appearance of lipid globuli and their formation is related to the endoplasmic reticulum of rough type (ERr). The lipid globules or pro-orbicules disappear in the mature Ubisch bodies, and the places that they occupied remain free of contents or with pollen-kitt.

Effect of Maternal Thyroxine Treatment on the Postnatal Development of Brain-derived Neurotrophic Factor-containing Purkinje Cell in Fetal Alcohol Effects in the Rat Cerebellum / 체질인류학회지

Maternal alcohol abuse is thought to be the common cause of mental retardation. Even moderate maternal alcohol consumption may produce fetal alcohol effects with behavioral and learning difficulties, if the drinking is associated with malnutrition. Especially, continuous alcohol consumption during critical period of brain development is very likely to produce fetal alcohol effects. The aims of this study are to investigate whether exogenous thyroxine treatment to alcohol -fed dams may ameliorate the detrimental effects of alcohol on the postnatal development of BDNF -containing Purkinje cell of the cerebellar cortex of the offspring. The morphological features of the growth and maturation were observed at 0, 7, 14, 21, 28 postnatal days via immunohistochemistry. In addition, electron microscopic finding of BDNF -containing Purkinje cell at P14 was also examined. Time -pregnant rats were divided into three groups. Alcohol -fed group received 35 calories of liquid alcohol diet daily from gestation day 6; control pair -fed group was fed a liquid diet in which dextrin replaced alcohol isocalorically; alcohol +/-T4 group received 35 calories liquid alcohol diet and exogenous thyroxine subcutaneously. As a result, a similar developmental pattern of BDNF -immunoreactive Purkinje cells was observed in control pair - fed and alcohol+/-T4 group on and after P14. These cells of alcohol -fed group showed immature features. Single -layer arrangement of these cells in alcohol -fed group was not completely achieved throughout postnatal life. Electron microscopic observations of BDNF -immunoreactive Purkinje cells at P14 revealed large nucleus, small cytoplasm, small amount of ribosomal collection and rudimentary cytoplasmic organelles in alcohol -fed group. The morphology of BDNF -immunoreactive Purkinje cell in alcohol +/-T4 group was similar to that in control pair -fed group. It was characterized by numerous short segments of rough endoplasmic reticulum, many of which showed a tendency of parallel alignment that suggested an attempt at Nissl body configuration. The cytology of Golgi complexes was also found within the cytoplasm in perinuclear location. Those observed differences of postnatal maturation patterns between alcohol -fed and alcohol +/-T4 group may indicate the beneficial effects on the postnatal development of BDNF -containing Purkinje cells in cerebellar cortex in the pups of thyroxine -treated alcohol -exposed dams. These results suggest that the increase of BDNF synthesis during early postnatal life caused by maternal administration of exogenous thyroxine may ameliorate fetal alcohol effects as a result of the dysthyroid state following maternal alcohol abuse.

An Immuno-Electron Microscopic Study for Apoptosis and Expression of p53 on the Synovium of the Osteoarthritic Knee in Human / 대한해부학회지

This study was designed to observe the apoptosis and expression of p53 in the osteoarthritic synovial membrane compared with normal synovial membrane of human. The collected normal and osteoarthritic synovia were dissected and fixed for two hours (in 4% paraformaldehyde and 0.1% glutaraldehyde solution). In this study, TUNEL staining and immunocytochemical gold labeling techniques were used. In the immunocytochemical gold labeling techniques, primary antibodies which was to be monoclonal mouse anti-p53 were used. Donkey anti-mouse IgG tagged with 6 nm colloidal gold particles was used as the secondary antibody. The tissues were observed under JEOL 1200 EX-II transmission electron microscope. The results were as follows. 1. On TUNEL staining, normal synovium were not seen TUNEL positive signal cells. But, in the osteoarthritic synovium, few TUNEL positive cells were seen in synovial membrane and subsynovial layers. 2. On the transmission electron microscopic observation, normal synovium had 1~3 synovial cell layers, which had phagocytic synovial cells and secretory synovial cells. The osteoarthritic synovium had 2~5 synovial cell layers, which consisted with abnormally proliferated secretory synovial cells. These cells had heterochromatin in nucleus and well developed endoplasmic reticulum in the cytoplasm. 3. On the normal synovium of the human knee joint, p53 positive cells were not identified. But, in the osteoarthritic synovium of the human knee joint, p53 positive cells were identified. These cells were recognized secretory synovial cells and apoptotic cells. In the secretory synovial cells, the distributions of p53 were mitochondria and rough endoplasmic reticulum. In the apoptotic cells, p53 were marked on rough endoplasmic reticulum, which showed secretory synovial cells. On the basis of above findings, it is obvious that osteoarthritic synovial membrane has identified the apoptotic cells compared with normal synovium. These apoptotic cells might be identified as mainly secretory synovial cells and a few phagocytic synovial cells. The immunogold of p53 was marked at rough endoplasmic reticulum and in nucleus of apoptotic cells. Apoptosis in the osteoarthritic synovium seemed to be developed through p53 negative dependent pathway.

Electron Microscopic Autoradiographic Study on the Secretion of Atrial Specific Granules from Atrial Cardiocytes in Dehydrated Rats / 대한해부학회지

This study was performed to investigate for the effect of dehydration on the synthesis, secretion and secreted pathway of atrial specific granules contained ANP by electron microscopic autoradiography. Male Sprague-Dawley rats, body weigh of about 50 g (range 47 to 53 g), were divided into control, 1 day dehydration and 3 days dehydration groups. Each group was divided into four groups according to sacrificed time on 20 min, 60 min and 240 min after the injection of L-leucine 3 H. Tissues of the right atrium obtained from animals were processed for typical electron microscopic procedure, and then embedded in Epon 812. Ultrathin sections were followed for electron microscopic autoradiographic method. Atrial specific granules were various in size, and some granules had a lower electron densities and indistinct granular membrane in the dehydration groups compared with the control group. In the electron microscopic autoradiographs of atrial wall, silver grains indicated by means of the positions of labelled L-leucine 3 H over the cell inclusion included atrial specific granules, cell organelles, intercellular spaces and blood vesseles. In the control group, high specific radioactivity was observed in the Golgi apparatus at 20 min, and in the rough endoplasmic reticulums and atrial specific granules at 60 min after the injection of L-leucine3H. And high level of radioactivities were observed in the cell membranes and blood vesseles at 240 min after the injection of L-leucine3H. In the 1 day and 3 days dehydration groups, radioactivities of Golgi apparatus, atrial specific granules, cell membranes and intercellular spaces were high level at 20 min, and radioactivities of rough endoplasmic reticulums and blood vesseles were high level at 60 min after isotope injection. Stored atrial specific granules were increased to 34.1% in the 1 day dehydration group, 27.4% in the 3 days dehydration group compared with the control group. In the 3 days dehydration group, newly formed granules increased 85.02% at 20 min, but those decreased rapidly to 36.87% at 60 min, 20.45% at 240 min after the injection of L-leucine3H in atrial cardiocytes. This results suggest that total ANP increased rapidly in the atrial cardiocytes, and newly formed ANP secreted rapidly into the intercellular space in the condition of dehydration, and ANP from atrial cardiocytes remain in intercellular space for dehydration period.

Study on expression of glycosaminoglycan in adenoid cystic carcinoma

Adenoid cystic carcinoma is malignant tumor in salivary gland, and its behavior is very invasive. Of all malignant tumor adenoid cystic carcinoma is occured in frequency of 4.4% in major salivary gland, and 1.29% in minor salivary gland. Histopathologically, adenoid cystic carcinoma is characterized by a cribriform appearance, and tubular form and solid nest type tumor can be seen. The tumor cell structure composed of modified myoepithelial cell, and basaloid cell. Extracellular matrix of this tumor cell contains variable ground substance with basement membrane component. Basement membrane matrix composed of collagen fibers, glycoproteins, proteoglycans, and its function is well known that it participate in differentiation, proliferation, and growth of tumor cell. Basement membrane molecule is essential for invasion of peripheral nerve, blood vessel, skeletal muscle in tumor cell of adenoid cystic carcinoma. In many studies, the tumor cell of adenoid cystic carcinoma containing modified myoepithelial cell participate in synthesis of proteoglycan. In this study, tissue sample of adenoid cystic carcinoma of human salivary gland were obtained from 15 surgical specimen, and all specimen were routinely fixed in 10% formalin and embedded. Serial 4-micrometer thick sections were cut from paraffin blocks. the histopathologic evaluation was done with light microscopy. And, the immunohistochemical staining, characteristics of glycosaminoglycan were observed. For biochemical analysis of glycosaminoglycan, isolation of crude glycosaminoglycan from tumor tissue and Western bolt analysis were carried out. With transmission electomicroscopy, tumor cell were observed. Biologic behavior of adenoid cystic carcinoma was observed with distribution and expression of basement membrane of glycosaminoglycan in tumor cells, The results obtained were as follows: 1. In immunohistochemical study, chondroitin sulfate is postively stained in tumor cell and interstitial space, dermatan sulfate is weakly stained in ductal cell. But keratan sulfate is negatively stained. 2. In immunohistochemical study, heparan sulfate is strong positive stained in tumor cell and basement membrane, especially in invasion area to peripheral nerve tissue. 3. In transmission electromicroscpic view, the tumor cells are composed modifed myoepithelial cells, and contains many microvilli and rough endoplasmic reticulum. 4. In Western blot analysis, the expression of glycosaminoglycan is expressed mostly in heparan sulfate. From the results obtained in this study, tumor cell of adenoid cystic carcinoma is composed modified myoepithelial cell, and glycosaminoglycan of basement membrane molecule of heparan sulfate and chondroitin sulfate mostly participate in the development and invasiveness of adenoid cystic carcinoma by immunohistochemical study and western blot analysis.

Bone Formation by rhBMP-7 Transduced HEK 293 Cells in Nude Mouse / 영남의대학술지

To induce bone formation at ectopic site by tissue engineering and gene therapy, we transplanted collagen sponges containing rhBMP-7 transduced HEK 293 cells in the hypodermis of nude mice. Bone formation was investigated by histological and electron microscopic method at 3, 6, and 9 weeks after transplantation. At 9 weeks after transplantation, eosinophilic bony tissue was observed in the implanted collagen sponge and was confirmed as bone tissue by Von Kossa stain. In the transmission electron microscopic observation, the cells in newly formed bone tissue had eccentrically located nucleus and well developed rough endoplasmic reticulum (rER). Therefore, the cells were evaluated as osteoblasts. Those results suggest that it is possible to form a bone tissue in the ectopic site by transplantation of rhBMP-7 transduced HEK 293 cells. This will be contributed to push more advanced gene therapy for bone formation. However, the HEK 293 cell is unable to apply to the clinical gene therapy. Therefore it is worth to find more compatible cells for clinical application. In addition, collagen sponge is considered as an excellent scaffold and/or carrier for gene therapy and a good biomaterial for tissue engineering.

Bone Formation by rhBMP-7 Transduced HEK 293 Cells in Nude Mouse / 영남의대학술지

To induce bone formation at ectopic site by tissue engineering and gene therapy, we transplanted collagen sponges containing rhBMP-7 transduced HEK 293 cells in the hypodermis of nude mice. Bone formation was investigated by histological and electron microscopic method at 3, 6, and 9 weeks after transplantation. At 9 weeks after transplantation, eosinophilic bony tissue was observed in the implanted collagen sponge and was confirmed as bone tissue by Von Kossa stain. In the transmission electron microscopic observation, the cells in newly formed bone tissue had eccentrically located nucleus and well developed rough endoplasmic reticulum (rER). Therefore, the cells were evaluated as osteoblasts. Those results suggest that it is possible to form a bone tissue in the ectopic site by transplantation of rhBMP-7 transduced HEK 293 cells. This will be contributed to push more advanced gene therapy for bone formation. However, the HEK 293 cell is unable to apply to the clinical gene therapy. Therefore it is worth to find more compatible cells for clinical application. In addition, collagen sponge is considered as an excellent scaffold and/or carrier for gene therapy and a good biomaterial for tissue engineering.

Development and Migration of Megakaryocyte during Hepatic Hemopoiesis in Human Fetuses / 대한해부학회지

Liver tissuses obtained from 5 human fetuses between 11 weeks and 23 weeks of gestation during the high activity of hepatic hemopoiesis were observed with transmission electron microscope using continuous series of thin sections. The objective of present study was to evaluate ultrastructures of megakaryopoietic cells, the migration of extravascular megakaryocyte into the sinusoidal lumen and the relevence between a migrated megakaryocyte and a Kupffer cell. Immature megakaryocytes were usually observed between growing hepatic laminae and within hepatic sinusoids. A megakaryoblast contained numerous polyribosomes, rather large mitochondria, short tubular elements of rough endoplasmic reticulum and small granules. Moreover, demarcation tubules and a few small specific granules were observed in immature megakaryocytes. The nucleus was mononuclear but frequently indented. With maturation, the nuclei were multilobulated. In the cytoplasm, in contrast to the decrease in polyribosomes and rough endoplasmic reticulum, the numerous specific granules and well -developed demarcation membrane system were predominant. Thereafter cytoplasmic zonation was observed clearly in maturing and mature megakaryocytes. Some megakaryocytes passed through the sinusoidal lining epithelium and into the hepatic sinusoids. The cell to cell interaction was often found as adhesion between migrated megakaryocyte and Kupffer cell, and erythroblasts within megakaryocyte (emperipolesis). These results suggest that intravascular megakaryopoiesis in addition to extravascular megakaryopoiesis occurs to produce platelet during the human fetal liver.