The programed cell death of the cutaneous epithelial tissue during tail regression stages in anuran tadpoles of the blackspotted frog, Rana nigromaculata were visualized by the histochemical and transmission electron microscopic techniques. Metamorphotic changes in the tail regression during the period of the Shumway stage number 31 to 33 are characterized by the disappearance of mucous layer and formation of compound epithelium through cutaneous thickening. Following the TUNEL (terminal deoxynucleotidyl transferase-mediated biotinylated d-uridine triphosphate nick end labeling) staining technique, the apoptotic cells were detected at the distal region of the tail skin initially, but they can be seen at the proximal region according to their following development. It has been also revealed that the number of the TUNEL-positive cells gradually increased from apical to basal direction of the epithelial layers during the tail regressing stages. Following the TEM observation, the early apoptotic cells shown in the epithelium demonstrated condensation and margination of the chromatin material at the nuclear periphery. Another epithelial apoptotic cells were shown nuclear fragmentation, membrane blebbing and cytoplasmic condensation. Following the process of the apoptotic degradation, well preserved organelles and nuclear fragments can be identified in the cytoplasm of lysosome-rich cells, however they soon reduced to lysosomal residual bodies through the progressive degradation.
The purpose of this study is to describe the cellular characteristics of endometrial hyperplasia without/with atypia in cervical smears. These cellular features were compared with those of normal endometrium and endometrial carcinoma. We reviewed 265 cervical smears : 64 normal proliferative endometrium, 118 endometrial hyperplasia without atypia, 21 endometrial hyperplasia with atypia, and 62 endometrial adenocarcinoma. Of these smears, 72(27.2%) smears which had diagnostic endometrial epithelial cells were selected for this study. The cytologic abnormalities about cellularity, background, changes in cellular architecture, alterations in nuclear size, anisokaryosis, chromatin pattern, nucleoli, cytoplasmic vacuoles, and mitosis were observed. Nuclear enlargement(1.6 to 2 times of the nucleus in the intermediate squamous cell) and anisokaryosis(${\geq}$2 fold in size variation) were highly suggestive of endometrial hyperplasia without/with atypia. The nuclei from endometrial hyperplasia with atypia were more coarsely granular in chromatin patterns than hyperplasia without atypia(33.3% vs 3.4%). Micronucleoli were observed in all endometrial conditions, but the presence of macronucleoli were more suggestive of hyperplasia with atypia(22.2%) and adenocarcinoma(55%). The changes in cellular architecture(loss of polarity, uneven internuclear distance, overlapping and loose arrangement) were seen in hyperplasia with atypia and adenocarcinoma. Characteristically, bloody background was seen in endometrial hyperpiasia, and cellular detritus or granular proteinaceous material was only observed in endometrial adenocarcinoma. Mitoses were also observed in adenocarcinoma. In conclusion, although there is no single parameter useful for the cytologic differential diagnosis of endometrial lesions, combined cytologic evaluation can be used to diagnose hyperplasia cytologically.
cis-Dichlorodiammineplatinum (II) (cis-Platin), a metallic compound, has widely been used as an effective anticancer chemotherapeutic agent. The precise mechanism of action of this agent is still unknown, but it is postulated that cis-Platin may act on the cancer cell like bifunctional alkylating agents. Although this agent is very beneficial to the patients with cervical cancer, germinoma of testis, neuroblastoma and others, it may also damage to the normal cell so that many side effects; severe hemorrhagic enterocolitis, bone marrow depression, renal damage and liver damage will develope. This experiment has been undertaken to pursue the cytotoxic effects of the cis-Platin on the ultrastructures of the interalveolar septum in the mouse lung. A total of 55 healthy male mice of ICR strain were used as experimental animals and divided into 5 mice of normal control group and 50 mice of cis-Platin treated group. The mice of cis-Platin treated group were sacrificed by carotid exsanguination at 6, 12, 24 hours, 3 days and 7 days after intraperitoneal injection of 6.0 mg of cis-Platin ($Abiplatin^R$ Abic Co. Ltd.) per kg of mouse body weight. The specimen obtained from the lower lobe of left lung were sliced into $1mm^3$ and prefixed with 2% glutaraldehyde -2.5% paraformaldehyde solution prepared with Millonig's phosphatae buffer solution (pH 7.4) at $4^{\circ}C$ for 3-4 hours. After postfixation with 1% osmium tetroxide solution all specimens were embedded in Epon 812. Ultrathin sections about $600-800{\AA}$ in thickness were stained with uranyl acetate and lead citrate and observed with Hitachi-600 electron microscope. The results obtained were as follows: 1. Local swellings with increase of electron density and number of pinocytic vesicles in the cytoplasms of the type I pneumocyte and endothelial cell of the blood air barrier in interalveolar septum of cis-platin treated mice were observed. 2. Cisternae of rough endoplasmic reticulum were dilated and sacculated in association with detachment of membrane bound ribosomes of the type II pneumocyte in interalveolar septum of cis-Platin treated mice. 3. Swollon mitochondria with uneven electron density of their matrix were observed in the type II pneumocyte of interalveolar septum in the cis-Platin treated mice. 4. The lamellae of lammelar bodies in type II pneumocyte of interalveolar septum in cis-Platin treated mice were devoided or transformed into homogeneous electron dense material. It is consequently suggested that cis-Platin would induce the cellular edema of type I pneumocyte and endothelial cell, and degenerative changes of cytoplasmic organelles of the type II pneumocyte in the interalveolar septum of the mouse lung.
The fine structure of the aggregate glands-one of the capture thread producing organs-in the orb web spider, Nephila clavata L.Koch, is studied with light and electron microscopes. Gluey capture threads or sticky spirals of the orb web are originated from the silks of two flagelliform glands and four aggregate glands which are connected to the posterior spinnerets, and the arrangement fo their spigots(large spinning tubes) shows a charaterstic form called "triad". The aggregate galnd is composed of large and multilobed secretory portion and thick excretory duct surrounded by large irregular nodules. The excretory duct of the aggregate galnds basically consists of three superposed types of cells which are inner columnar epithelium, nodule-forming cells and outer connestives. The cuticles of the proximal duct near the secretory portion are composed of endocuticle and exocuticle, whereas ghe distal duct near the spinning tubes has a electron lucent subcuticle which had the functions of water removal and orientation of silk fibers. In the cytoplasmic process of the large and irregular nodule-forming cells surrounded by invaginations of the plasma membranes, numerous mitochondria and glycogen particles are contained. The maturational level of the nodule cells is perceived from the appearence of these cell inclusions. The secretory portion of the glands which porduce the secretory silk material shows two layers of the cells which are simple cuboidal epithelium and several connective layers. In the cytoplasm of the glandular epithelial cell, rough endoplasmic reticulums are well developed, and two types of secretory granules are observed. Between the adjacent epithelial cells, specialized septate junctions are formed along the plasma membranes.membranes.
STATEMENT OF PROBLEM: A biochemical approach for surface modification has offered an alternative for physicochemical and morphological methods to obtain desirable bone-implant interfaces. PURPOSE: The purpose of the present study was to investigate cell responses to poly (D,L-lactide-co-glycolide) (PLGA)/$1{\alpha}$,25-(OH)$_2D_3$ coating with reference to cellular proliferation and differentiation in vitro. MATERIAL AND METHODS: 96 titanium discs were fabricated and divided into four groups. Group 1 was anodized under 300 V as control. Group 2, 3 and 4 were anodized then coated with 3 ml PLGA/$1{\alpha}$,25-(OH)$_2D_3$ solutions. Amount of the solutions were 2 ul, 20 ul and 200ul respectively. The osteoblast-like Human Osteogenic Sarcoma (HOS) cells were seeded and cultured for 1, 3 and 7 days. MTSbased cell proliferation assay and ALPase activity test were carried out. RESULTS: PLGA nanoparticles were observed as fine, smooth and round and HOS cells attached to the anodized surfaces through strand-like and sheet-like filopodia. After 3 days of culture, the dendritic filopodia were exaggerated and sheet-like cytoplasmic projections covered the coated titanium surfaces. After 3 days of culture, all of the groups showed increased cellular proliferation and the lowest proliferation rate was measured on group 2. Higher amount of incorporated $1{\alpha}$,25-(OH)$_2D_3$ (Group 3 and 4) improved cellular proliferation but the differences were not significant statistically (P > .05). But they increased the rate of ALP activities than the control group at day 3 (P < .05). CONCLUSION: Biodegradable PLGA nanoparticles incorporated with vitamin D metabolite positively affected proliferation and differentiation of cells on the anodized titanium surface.
Park, Chun-Geon;Bang, Kyong-Hwan;Lee, Seung-Eun;Cha, Moon-Seok;Sung, Jung-Sook;Park, Hee-Woon;Seong, Nak-Sul
Korean Journal of Medicinal Crop Science
/
v.9
no.4
/
pp.251-258
/
2001
Medicinal plant extracts including Rubus coreanus, Sanguisorba officinalis, Eriobotrya japonica, Prunus mume, Crataegus pinnatifida, Rosa leavaigate Prunus persica, Prunus japonica var. nakaii and Spiraea blumei were prepared for the test of antibacterial activity. Tryptic soy broth (TSB) containing $0{\sim}10mg/ml$ of medicinal plant extracts was inoculated with $10^6$ cells/ml of Staphylococcus aureus and incubated at $35^{\circ}C$ for 24 hours. The plate counting method and clear zone test were used to test inhibitory effect of the extracts. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) was derived from the survival curves of S. aureus. The order of antibacterial activities of medicinal plant extracts on the S. aureus was Rubus coreanus > Sanguisorba officinalis > Eriobotrya japonica > Prunus mume > Crataegus pinnatfida. Minimum inhibitory concentration of Sanguisorba ofEcmalis on the Staphylococcus aureus was 2.5mg/ml and minimum bactericidal concentration of Rubus coreanus was 1.0%. Inhibition zone of Rubus coreanus, Sanguisorba officinalis, Eriobotrya japonica, Prunus mume, and Crataegus pinnatifida was 16.5mm, 14.3mm 11.0mm, 14.0mm and 12.7mm, respectively. The morphology of S. aureus cells treated with medicinal plant extracts showed damage of cell wall and cytoplasmic membrane. Severely damaged cells of S. aureus lost electron dense material and cytoplasm. This result suggests that medicinal plant extracts can be used as an effective natural antibacterial agent in food.
The regeneration and differentiation of the cutaneous pigment system in the goldfish, Carassius auratus during the wound healing process were studied with high magnification electron microscope. The cutaneous pigment cells of the normal tissues were composed of three kinds of dermal chromatophores-xanthophores, leucoiphores and melanophores. While xanthophores contain two kinds of pigment granules-pterinosomes and carotenoid vesicles, leucophores and melanophores contain amorphous pigment granules (leucosomes) and oval shaped electron dense melanin pigment granules (melanosomes) respectively. After injury, primary wound healing responses being carried out by migration of epidermal cells and hemocytes spreading over the wound surface at the day of wounding. And at the time of primary wound closure, 5 to 7 days after wounding, rER rich cells-presumably common precursors of dermal chromatophores-immigrated into the wound area. First redifferentiated chromatophores appeared 3 weeks after wounding. Pigment granules of the chromatophores were emerged from the cytoplasmic Golgi complex via rough endoplasmic reticulum. Pinocytotic vesicles which associated with accumulation of pigment material, appeared only at the inner surface of the chromatophores adhering to the rER rich cells, characteristically. The differentiation of each chromatophore in addition to integumental wound repair were accomplished within 4 weeks after wounding at most cases, however the total numbers and densities of these repaired chromatophores still primitive state. Moreover, It has been revealed that complete repair of chromatophores at wounded tissues from burns requirs more than 3 months in normal environment.
In order to understand the action mechanism of polyhexamethylene biguanide (PHMB) to the cyst of Accnthcnloebc on the morphological basis, the cysts of four corneal isolates of Acanthanoebc were treated with minimal cysticidal concentration (MCC) of PHMB and their ultrastructural changes were examined by transmission electron microscopy. The most striking change of cysts treated with PHMB compared with normal cysts was the shrinkage of intracystic amoebae, which resulted in the separation of the plasma membrane of intracystic amoeba from endocystic wall. Subplasmalemmal lipid droplets became irregularly shaped . In severely damaged cysts, cytoplasm was aggregated and organelles were severely deformed. Cytoplasmic materials were leaked out through the damaged plasma membrane. Most cysts showed aggregation of nuclear chromatin material. Number of mitochondrial cristae was also reduced. Ecto- and endo-cystic walls were relatively well tolerated. Findings in the present study revealed that PHMB affected mainly on plasma membrane, but lesser on organellar membrane of intracystic amoeba. It seemed likely that PHMB might kill cystic forms of Accnthamoebc by similar mechanism in which this environmental biocide can damage the cell wall of Escherichia coli by binding with acidic phospholipids.
Kim Young-Min;Lee Chang-Yun;Yang Kyung-Hun;Rho Young-Soo;Park Young-Min;Lim Hyun-Jun
Korean Journal of Head & Neck Oncology
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v.14
no.2
/
pp.236-243
/
1998
Objectives: Local invasion of the thyroid cancer that is invasion of the upper aerodigestive tract, neurovascular structures of the neck and superior mediastinum, is infrequent and comprises of 1-16% of well-differentiated thyroid cancer. However the proximity of the thyroid gland to these structures provides the means for an invasive cancer to gain ready access into theses structures and when invasion occurs, it is the source of significant morbidity and mortality. So locally invasive thyroid cancer should be removed as much as possible, but still much debates have been exist whether the surgical method should be radical or conservative. This study was desinged to evaluate the clinical characteristics and the surgical treatment of the locally invasive thyroid cancer. Material and Methods: At the department of otorhinolaryngology of Hallym university, 10 patients diagnosed as locally invasive thyroid cancer among the 81 patients treated for thyroid cancer between 1991 to 1997 were retrospectively evaluated. Results: Of the 10 patients, 3 patients had histories of previous surgical treatment with or without radiation or radioactive iodine therapy. The site of invasion of thyroid cancer were trachea(7 cases), recurrent laryngeal nerve(5 cases), mediastinal node(5 cases), esophagus(3cases), larynx(3cases), carotid artery(3 cases), pharynx(l case), and other sites(4 cases). The operation techniques included 1 partial laryngectomy and 1 partial cricoid resection, 2 shavings and 3 window resections of the trachea, 1 sleeve resection of the trachea with end-to-end anastomosis and 1 cricotracheoplasty for tracheal invasion, 2 shavings and 1 partial esophagectomies for esophageal invasion, and 1 wall shaving and 2 partial resections with $Gortex^{\circledR}$ tube reconstruction for carotid artery invasion, and so on. Conclusions: These data and review of literature suggest that the surgical method should be perfomed on the basis of individual condition and complete removal of all gross tumor with preservation of vital structures whenever possible will offer a good result.
Background: Due to the paucity of suitable donor organs for lung allotransplantation, a number of techniques have been developed to improve the lung preservation. Ultrastructural studies of the morphologic changes of the flushing, preservation and reperfusion injury in donor lungs have rarely been reported. Methods: Adult dogs (n=46) were matched as donors and recipients for the single lung transplantation. The donor lungs were preserved after flushing with preservation solution and transplanted after 20-hours of preservation at $10^{\circ}C$. Ultrastructural features of the lung were examined after flushing, preservation and 2 hours after lung transplantation (reperfusion) respectively. Results: Electron microscopy after flushing showed focal alveolar collapse and mild swelling of type I epithelial cells. After preservation both type I epithelial cells and endothelial cells were swollen and destroyed focally. The endothelial cells showed protrusion of tactile-like structures into the lumina, blebs or vacuoles of the cytoplasm After reperfusion the lung tissue showed fibrin material in the alveoli, prominent type I epithelial cell swelling with fragmented cytoplasmic debris and marked endothelial cell swelling with vacuoles or tactile-like projections. The alveolar macrophages showed active phagocytosis. Scanning electron microscopic examination of the pulmonary parenchyma showed focally alveolar collapse and focal consolidation after the preservation and more prominent changes after the reperfusion procedure. The lungs preserved with low potassium dextran glucose solution, with additional prostaglandin $E_1(PGE_1)$ and verapamil(VP) showed relatively well preserved ultrastructures compared with those which were preserved with modified Euro-Collins or University of Wisconsin, and with additional $PGE_1$ and/or VP. Conclusion: The ultrastructural changes associated with flushing were mild in severity, the donor lungs were injured during the preservation, and further damage was occurred during the reperfusion. The reperfusion injury resulted in prominent pulmonary parenchymal alterations with a pattern of acute lung injury.
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