In an attempt to evaluate the cardiotoxicity of bupivacaine, beating rate, tetrazolium MTT and lactate dehydrogenase activity were investigated in the medium containing bupivacaine for 24 hours after neonatal rat myocardial cells were cultured for 72 hours. Light and electron microscopic studies were also carried out. The results were as follows ; 1) Beating rate decreased dose-dependently, and beating cells were not observed over 10(-4) M concentration of bupivacaine. 2) MTT50 value was 0.32 ug/ml (1,000 uM). 3) The amount of lactate dehydrogenase released into the medium was 192% of control cells at 10(-3) M concentration of bupivacaine. 4. In light microscopy, myocardial cells were decreased in number dose-dependently, and showed a few cytoplasmic processes and lots of granules in cytoplasm at 10 M concentration of bupivacaine. 5. Electron microscopy of bupivacaine-treated cells showed smooth endoplasmic reticulum, destruction of mitochondria and Golgi apparatus and increase of vacuoles and dense bodies. It also showed dilatation of rough endoplasmic reticulum and loss of myofibrils. These results suggest that high concentration of bupivacaine (> or = 10(-4) M) induee remarkable toxicity on cultured rat myocardial cells.
Animals ; Bupivacaine* ; Cytoplasm ; Dilatation ; Endoplasmic Reticulum, Rough ; Endoplasmic Reticulum, Smooth ; Golgi Apparatus ; L-Lactate Dehydrogenase ; Microscopy ; Microscopy, Electron ; Mitochondria ; Myofibrils ; Rats* ; Vacuoles

In this study, to understand the pathogenesis of new rabbit hemorrhagic disease virus (RHDVa) serotype, we carried out to administrate RHDVa to rabbits, and to examine sequential electron microscopic changes and relationship between pathogenesis and apoptosis. TUNEL-positive cells began to be observed from 24 hours after inoculation (HAI) and the number of positive cells was slightly increased with the course of time. Whereas marked increase of positive cells was seen in the liver from the rabbits died acutely. Typical viral particles with cup-like projections and a diameter of 30~40 nm were detected in homogenized liver samples and tissues at 36 and 48, and 48 HAI, respectively. Ultrastructurally, glycogen deposition was observed from the first stage of hepatocellular degeneration by RHDVa infection and then, swelling and disruption of cristae of mitochondria by viral particles, swelling of smooth endoplasmic reticulum, vacuoles and vesicles were detected. Condensation, margination and fragmentation of chromatin were observed in degenerative hepatocytes at 36 and 48 HAI, indicating apoptotic bodies. These data offer that hepatocytic apoptosis by RHDV infection could be closely related with mitochondrial impairment in the hepatocytes.
Apoptosis ; Chromatin ; Electrons ; Endoplasmic Reticulum, Smooth ; Glycogen ; Hemorrhagic Disease Virus, Rabbit ; Hepatocytes ; Liver ; Mitochondria ; Rabbits ; Sprains and Strains ; Vacuoles ; Virion

Ciliogenesis was investigated in the tracheal epithelium of human fetus at mid trimester of gestation (15~22 weeks), and the substructure of basal body was studied with serial, cross sections. The ciliogenic cells were long columnar cells with an electron -lucent cytoplasm, and contained rich free ribosomes and smooth endoplasmic reticulum. Apical cytoplasm of these cells contained various structures related to ciliogenesis including fibrous granules, procentrioles, centrioles and basal bodies. Basal bodies were located near apical plasma membrane and had basal foot and striated rootlets. In cross section, alar sheets appeared at transitional area between distal portion of basal body and axoneme, and basal foot at distal portion of basal body. Alar sheets arouse from each peripheral triplets of basal body and projected radially clockwise in apex to base view. Basal foot was a cone shaped structure with cross striation which base attached to two or three of the peripheral triplet sets and apex converged to basal foot cap. Three dimentional reconstruction by serial cross section of the basal body showed a structural relationship of alar sheets and basal feet with basal body. By immunohistochemistry, alpha -tubulin label was seen in both basal and surface ciliated cells, and gamma-tubulin label was seen in the apical region of surface cilated cells. These results indicate that ciliogenesis of tracheal epithelium of human fetus is performed mainly by acentriolar ciliogenesis, and suggest the ciliogenesis and ciliary movement at mid trimester of gestation are active.
Axoneme ; Basal Bodies ; Cell Membrane ; Centrioles ; Cytoplasm ; Endoplasmic Reticulum, Smooth ; Epithelium* ; Fetus* ; Foot ; Humans* ; Immunohistochemistry ; Pregnancy ; Ribosomes ; Triplets ; Tubulin

Hyperosmotic agents such as mannitol are widely used in ophthalmology to lower intraocular pressure as a short-term or emergency method. The mechanism of action of these agents is not fully understood, but probably relates primarily to a reduction in vitreous volume. There are other theories of hypotensive meechanism such as hypothalamic-neural theory and altered epithelium theory. The author performed this animal experiment for the eletronmicroscopic study of ciliary epithelium after the intravenous mannitol injection. Five healthy adult male albino rabbits weighing 2.5 kg were used in this experiment. Four rabbits were administered 25 ml(2 gm/kg) of 20% mannitol and the other one was given 25 ml of normal saline as a control through ear vein within 5 minutes each. The mannitol group was enucleated 10, 20, 40 and 80 minutes after injection and the control one was enucleated 20 minutes after injection. The enucleated eyes were opened and fixed in mixed solution of 2% paraformaldehyde, 3% glutaraldehyde and 0.2M Milonig's buffer. Small pieces consisting of ciliary body were excised, postfixed in 1% osmium tetroxide, dehydrated in ethylalcohol and embedded in Epon 812. Thin section were stained with toluidine blue for general histologic study and ultrathin sections stained with 4% uranyl acetate and 0.4% lead citrate were examined with a Hitachi H-600 transmission electronmicroscopy. The results were as follow: 1. The ciliary epithelium showed normal appearence 20 minutes after injection of normal saline and was composed of double layered epithelial cells. The tight juctions(zonulae occludens) were present between nonpigmented epithelial cells. The active Golgi apparatus, numerous mitochondria, rough endoplasmic reticulum and smooth endoplasmic reticulum were visible in the nonpigmented epithelial cells. The intercellular spaces were not dilated. 2. In mannitol group, no cellular necrosis was observed and cells were invariably present and apparently unaltered. 3. The intercellular spaces of ciliary epithelium began to dilate 10 minutes after intravenous mannitol injection, maximally dilated after 40 minutes and recovered after 80 minutes. 4. In view of the morphological changes of cytoplasmic organelles such as Goigi apparatus, the secretory function of nonpigmented epithelial cells after intravenous mannitol seemed to be inhibited maximally at 20 minutes and then recovered after 80 minutes. 5. In conclusion, the hypotensive mechanism of the mannitol on the ciliary epithelium was considered of secretory inhibition of nonpigmented epithelial cells besides diffusion by the osmotic gradient.
Adult ; Animal Experimentation ; Ciliary Body ; Citric Acid ; Cytoplasm ; Diffusion ; Ear ; Emergencies ; Endoplasmic Reticulum, Rough ; Endoplasmic Reticulum, Smooth ; Epithelial Cells ; Epithelium* ; Extracellular Space ; Glutaral ; Golgi Apparatus ; Humans ; Intraocular Pressure ; Male ; Mannitol* ; Mitochondria ; Necrosis ; Ophthalmology ; Organelles ; Osmium Tetroxide ; Rabbits ; Tolonium Chloride ; Veins

The aim of this study was to clarify the role of Kupffer cells in the mechanism of endotoxin-induced liver injury. The study on fine structure of Kupffer cells was performed after the injection of endotoxin. The endotoxin(Escherichia soli lipopolysaccharide 026: B6, 1.5mg/100 g of body weight) was intraperitoneally injected in Sprague-Dewley rats. Animals were sacrificed at 1/4, 1/2, 1, 2, 4, 8, 16, 24, 72 and 120 hours after the injection of endotoxin. Livers were extirpated and processed to be examined by light and electron microscopy. The results obtained were summerized as follows: Early changes observed in liver after endotoxin injection included the increased number and hypertrophy of Kupffer cells, infiltration of neutrophils and presence of fibrin thrombi within the sinusoids. The coritinuous increase of the Kupffer cells in number with hypertrophy, congestion and infiltration of inflammatory cells within the sinusoids were observed. Hepatocytes showed* fatty change and occasional necrosis. At 72 hours the congestion decreased. At 120 hours the number of Kupffer cells was increased, but the morphology of Kupffer cells became similar to that of the control group. The numbers and sizes of primary and secondary lysosomes and amount of euchromatin of Kupffer cells increased. Swellings and increase in number of mitochondria, Golgi complex, smooth endoplasmic reticulum, rough endoplasmic reticulum were evident. Microthrombi were present within the sinusoids. The swelling of rough endoplasmic reticulum and mitochondria, decrease of glycogen particles, fatty change, hypoxic vacuoles, pyknotic nuclei and occasional necrosis were observed in hepatocytes. At 72 hours the number of secondary lysosomes in Kupffer cells decreased. At 120 hours the morphology of Kupffer cells became similar to that of the control group. According to these results, it was postulated that the endotoxin was initially taken up by pinocytosis into Kupffer cells and degraded in secondary lysosomes of activated Kupffer cells. Kupffer cells may play an important role in the defense mechanism of liver during endotoxemia. The dysfunction of Kupffer cells and ischemia by sinusoidal microthrombi may cause liver injury.
Animals ; Endoplasmic Reticulum, Rough ; Endoplasmic Reticulum, Smooth ; Endotoxemia ; Estrogens, Conjugated (USP) ; Euchromatin ; Fibrin ; Glycogen ; Golgi Apparatus ; Hepatocytes ; Hypertrophy ; Ischemia ; Kinetics* ; Kupffer Cells* ; Liver ; Lysosomes ; Microscopy, Electron ; Mitochondria ; Necrosis ; Neutrophils ; Pinocytosis ; Rats* ; Vacuoles

Adriamycin has been widely used as an anticancer drug in the treatment of thyroid tumor, metastatic breast cancer, sarcoma and lymphoma. The antineoplastic effects of adriamycin result from its inhibitory action on the nucleic acid synthesis and the formation of reactive oxygen radicals by redoxcycling during its metabolic process. Adriamycin affects the normal cells of the patients and causes the undesirable side effects and the toxic effects. Thus, in this study we investigated the effect of dimethyl thiourea (DMTU), a hydroxyl radical scavenger, on the ultrastructural changes of the hepatocyte after the administration of adriamycin in the growing and adult rats. 36 Healthy male Sprague -Dawley adult rats (about 350 g) and 36 male rats at growing peroid (about 120 g) was used as experimental animals. Adriamycin (25 mg/kg) was administered intraperitoneally and DMTU (500 mg/kg) was administered intraperitoneally 30 minutes after the administration of adriamycin. The rats were sacrificed at 24 hours and 72 hours after the administration of adriamycin. A part of the liver from left anterior lobe was obtained and sliced into about 1 mm 3 . The specimens were prepared by the routine methods for electron microscopy. All preparations were stained with uranyl acetate and lead citrate and observed with Hitach -600 electron microscope. The results were as follows. 1. In the hepatocytes of the adult rat dilated, segmented and ribosome detached cisternae of rough endoplasmic reticulum, increased and dilated cisternae of smooth endoplasmic reticulum, damaged mitochondria and autophagic vacuoles were seen after the administration of adriamycin. The ultrastructural changes were progressive with the lapse of time. 2. In the hepatocyte of the growing rat dilated and ribosome detached cisternae of rough endoplasmic reticulum, changes of mitochondria, many lysosomes and the autophagic vacuoles were observed after the administration of adriamycin. 3. DMTU alone did not affect the ultrastructures of the hepatocytes in both growing and adult rats. 4. In the hepatocytes of growing and adult rats, dilated and ribosome detached cisternae of rough endoplasmic reticulum, dilated and increased cisternae of smooth endoplasmic reticulum, changes of mitochondria and autophagic vacuoles were seen after the combined administration of adriamycin and DMTU, but the degree of ultrastructural changes was
Adult* ; Animals ; Breast Neoplasms ; Citric Acid ; Doxorubicin* ; Endoplasmic Reticulum, Rough ; Endoplasmic Reticulum, Smooth ; Hepatocytes* ; Humans ; Hydroxyl Radical ; Liver ; Lymphoma ; Lysosomes ; Male ; Metabolism ; Microscopy, Electron ; Mitochondria ; Rats* ; Reactive Oxygen Species ; Ribosomes ; Sarcoma ; Thiourea ; Thyroid Gland ; Vacuoles

It is ordinarily accepted that the synthetic testosterone administration is the choice of treatment for the primary testicular dysfunction. But recently not a few medical scientists reported that the long-standing testosterone replacement is not without side effects. For that reason, we tried to investigate a possibility of in vivo transplantation of Leydig cells as a new biologic androgen substitution therapy. The Leydig cells procured from 6 week-old male Sprague-Dawley rat were auto- transplanted and the level of testosterone secretion and histostructural changes were observed.The results obtained are summarized as follows :1. For the selection of transplantation sites, compared to subcutaneous or scrotal counterparts, renal subcapsular and intraperitoneal transplant showed higher levels of testosterone and the number of transplanted cells was correlated with the level of measured testosterone. 2. Furthermore, if the Leydig cells were transplanted intraperitoneally after the uptake on synthetic collagen, testosterone levels were higher than the ones simply transplanted, resulting in 2.7 times higher at 3 months. 3. The activity of 125-I-hCG decreased 20 to 40% at each month after transplantation compared to the normal levels, but no statistical significance was noted among different periods. 4. Histologic examination revealed neovascularized capillaries and well demarcated uptake of sheet-like group of eosinophilic Leydig cells were observed at 4 weeks. But the evidence of destructive changes such as a focal inflammation with central dystropic ossification was noted after S month. On electron microscopy, the marked indentation of nucleus and presence of lipochrome pigment were seen and the number and size of smooth endoplasmic reticulum and mitochondria were reduced. In conclusion, testosterone output could be increased to the physiologic range by increasing the number of transplant cells or collagen uptake utilizing, etc. However the decrease of testosterone production after 3 months is believed to be a result of focal inflammation and degeneration which in turn caused the decrease in number of secreting Leydig cells and reduced activity of hCG receptors. Further effort is necessary on delaying or preventing the structural and functional decrement of Leydig cells.
Animals ; Capillaries ; Cell Transplantation* ; Collagen ; Endoplasmic Reticulum, Smooth ; Eosinophils ; Humans ; Inflammation ; Leydig Cells ; Male ; Microscopy, Electron ; Mitochondria ; Rats ; Rats, Sprague-Dawley ; Receptors, LH ; Testosterone ; Transplantation ; Transplants*

Leiomyosarcoma arising in the skin is rare tumor, and diagnosis usually is made microscopically. After local excision, these lesions recur in large proportion of pat ients. The authors herein report a 53-year-old male with leiomyosarcoma appeared in the skin of the right forearm and presenting as a dark reddish colored, 5*6cm in diameter, superficial ulcerated single firm nodule with intermittent pain. Histopathological examination showed poorly circumscribed tumor consisting of interlacing bundles of spindle shaped smooth muscle cells in the middle and lower parts of the dermis. The nuclei were hyperchromatic, large, vacuolated, and irregular in shape. Electron microscopic findings revealed cytoplasmic organelles such as rough endoplasmic reticulum and mitochondria of malignant smooth muscle cells in the paranuclear area, Characteristic subsarcoelmmal caveolae and dense plaque were noted and myofilaments were distributed in the peripheral cytoplasm. The tumor did not recur in 10 months' follow-up.
Caveolae ; Cytoplasm ; Dermis ; Diagnosis ; Endoplasmic Reticulum, Rough ; Follow-Up Studies ; Forearm ; Humans ; Leiomyosarcoma* ; Male ; Middle Aged ; Mitochondria ; Myocytes, Smooth Muscle ; Myofibrils ; Organelles ; Skin* ; Ulcer

Polyploid cells in the urinary sediments often give an erroneous clinical judgement in cases of post-pelvic irradiation follow-up, but their nature and evolution have remained unclarified. An experimental induction of polyploid cells in the transitional epithelium of the urinary bladder was carried out in Sprague-Dawley rats by administration of 3,000 rads in a single dose, and their sequential morphological changes were analysed under light and electron microscopes. 1. The acute post-irradiation changes of transitional epithelial cells were manifested with two consecutive phases of degenerative process ; the early lesion started to appear from the first day after irradiation and diminished partly at the 7th day; the later changes became enhanced progressively from the 2nd week and maximized at the 3rd week, but regressed thereafter . 2. The general histological alterations of the transitional epithelial cells in the acute stage were characterized by cytoplasmic vacuolization due to profound widening of intercellular cisternal spaces and dilatation of smooth endoplasmic reticulum aside from severe disruption of mitochondria and increase of lysosomes, especially in the superficial and intermediate cells, and by eventual outcome of cell death by nuclear pyknosis and karyorrhexis. 3. The polyploid cell change was demonstrated as a spectrum of the later alterations of acute irradiation injury to the basal layer cells, and appeared early from the 2nd week and regressed after the 4th week. 4. Based on their increased size and nuclear abnormalities, those polyploid cells exhibited features of both amitotic nuclei and cytoplasmic degenerative processes ultrastructurally, and in the acute phase the nuclear indentation and lobulation were associated with increased amount of heterochromatins and margination together with nucleolar enlargement and increase in number. 5. The above cells started to regress thereafter, being terminated by nuclear pyknosis and karyolysis, and numerical reduction of the polyploid cells was accompanied concomitantly with basal (reserve) cell hyperplasia of the remained epithelium. It is of the author's assumption that the polyploid cell phenomenon induced by irradiation onto the transitional epithelium of the urinary bladder is a transient manifestation of irradiated amitotic basal cells during the later phase of acute post-irradiation injury and is subsequently removed out by nuclear pyknosis and karyolytic processes.
Animals ; Cell Death ; Cytoplasm ; Dilatation ; Endoplasmic Reticulum, Smooth ; Epithelial Cells ; Epithelium* ; Follow-Up Studies ; Heterochromatin ; Hyperplasia ; Lysosomes ; Mitochondria ; Polyploidy* ; Rats* ; Rats, Sprague-Dawley ; Urinary Bladder Neoplasms ; Urinary Bladder*

OBJECTIVE: To investigate fertilization and embryo quality of dysmorphic mature oocytes with specific morphological abnormalities obtained from intracytoplasmic sperm injection (ICSI). METHODS: The fertilization rate (FR) and embryo quality were compared among 58 dysmorphic and 42 normal form oocytes (control 1) obtained from 35 consecutive ICSI cycles, each of which yielded at least one dysmorphic mature oocyte, performed over a period of 5 years. The FR and embryo quality of 441 normal form oocytes from another 119 ICSI cycles that did not involve dysmorphic oocytes served as control 2. Dysmorphic oocytes were classified as having a dark cytoplasm, cytoplasmic granularity, cytoplasmic vacuoles, refractile bodies in the cytoplasm, smooth endoplasmic reticulum in the cytoplasm, an oval shape, an abnormal zona pellucida, a large perivitelline space, debris in the perivitelline space, or an abnormal polar body (PB). RESULTS: The overall FR was significantly lower in dysmorphic oocytes than in normal form oocytes in both the control 1 and control 2 groups. However, embryo quality in the dysmorphic oocyte group and the normal form oocyte groups at day 3 was similar. The FR and embryo quality were similar in the oocyte groups with a single abnormality and multiple abnormalities. Specific abnormalities related with a higher percentage of top-quality embryos were dark cytoplasm (66.7%), abnormal PB (50%), and cytoplasmic vacuoles (25%). CONCLUSION: The fertilization potential of dysmorphic oocytes in our study was lower, but their subsequent embryonic development and embryo quality was relatively good. We were able to define several specific abnormalities related with good or poor embryo quality.
Abnormalities, Multiple ; Cytoplasm ; Embryonic Development ; Embryonic Structures* ; Endoplasmic Reticulum, Smooth ; Female ; Fertilization* ; Oocytes* ; Polar Bodies ; Pregnancy ; Sperm Injections, Intracytoplasmic ; Vacuoles ; Zona Pellucida