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

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

Prenatal development of the thoracic aorta of the human during the period ranging from gestation weeks 7 (C-R length 20mm) to 30 (C-R length 260mm) was examined by transmission electron microscopy and the following results were obtained. The early form of cuboidal or columnar endothelial cells at 7-9 weeks of gestation changed gradually to typical flat endothelial cells at 12-14 weeks of gestation. At 9 weeks of gestation, the mesenchymal cells begin to differentiate to myoblasts, which have small clusters of myofilaments with dense bodies and rough endoplasmic reticulum. And from 14 weeks the differentiating cells begin to form a parallel concentric lamellar structure. At 12th week of gestation, elastic fibers were first seen in subendothelial connective tissue and the intercellular spaces between smooth muscle cells. Elastic fibers appeared as small globular shape which composed of a central core of elastic and peripheral microfibrils. From this period the amount of elastic fibers and their aggregation increases gradually in both the subendothelial space and the intercellular spaces between smooth muscle cells. At 30th week of gestation, subendothelial elastic fibers almost completed the internal elastic lamina and also well formed elastic laminae were seen between the smooth muscle cells adjacent to endothelial cells. However, in the space between the smooth muscle cells near the adventitia the elastic lamina formation is delayed. In the adventitia elastic fiber were scanty but collagen fibers are abundant.
Adventitia ; Aorta, Thoracic* ; Collagen ; Connective Tissue ; Elastic Tissue ; Endoplasmic Reticulum, Rough ; Endothelial Cells ; Extracellular Space ; Fetus* ; Humans* ; Microfibrils ; Microscopy, Electron, Transmission ; Myoblasts ; Myocytes, Smooth Muscle ; Myofibrils ; Pregnancy

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

Histochemical and electron microscopic studies were made on the canal epithelium of the body segment of the rabbit epididymis and following results were obtained. 1) Acid phosphatase activity was marked in the canal epithelium. especially of the proximal body segment of the epididymis. Granules reactive to the acid phosphatase were present in both the above and below the nucleus 2) Electron microscopic finding: Canal epithelium of the body segment of the rabbit epididymis consisted of largely principal cells. Very few light and few basal cells. Principal cells were characterized by having slender microvilli (stereocilia). Luminal vesicles and vacuoles, extensive Golgi areas and many dense bodies in the supranuclear region, remarkable endoplasmic reticulum of mainly agranular type throughout the cytoplasm. Infranuclear cytoplasm contained often abundant mitochodria, many dense bodies and significant amount of granular endoplasmic reticulum. Light cells were characterized by having light cytoplasm, numerous vesicles, many vacuoles and dense bodies. Basal cells were present characterized by haying small nucleus, small number of cell organelles and rather clear cytoplasm From these results, it is suggested that the principal cell may have dual function of secretion and absorption and the light cell may engage mainly in absorptive function.
Absorption ; Acid Phosphatase ; Cytoplasm ; Endoplasmic Reticulum ; Endoplasmic Reticulum, Rough ; Epididymis* ; Epithelium* ; Male ; Microvilli ; Organelles ; Phenobarbital ; Vacuoles

It has been shown that mitochondria not only control their own Ca(2+) concentration ([Ca(2+)]), but also exert an influence over Ca(2+) signaling of the entire cell, including the endoplasmic reticulum or the sarcoplasmic reticulum, the plasma membrane, and the nucleus. That is to say, mitochondria couple cellular metabolic state with Ca(2+) transport processes. This review focuses on the ways in which the mitochondrial Ca(2+) handling system provides integrity and modulation for the cell to cope with the complex actions throughout its life cycle, enumerates some indeterminate aspects about it, and finally, prospects directions of future research.
Biological Transport ; Calcium Signaling ; Cell Membrane ; physiology ; Endoplasmic Reticulum ; physiology ; Mitochondria ; physiology ; Sarcoplasmic Reticulum ; physiology

When a glaucoma filtering operation fails, the causes may be categorized as intraocular, scleral, or extraocular. Some authors have implicated that extraocular causes of failure are due to closure of the external opening by connective tissue. This histologic investigation was undertaken to examine the surgical wounds of 8 experimental subjects including 4 albino and 4 pigmented rabbits. Under general anesthesia by pentothal sodium, a single punch sclerectomy was performed on each right eye. A trabeculectomy was performed on the left eye. Seven and ten days following surgery, globes were removed and immediately fixed in Karnovsky's fixation solution. Following fixation, globes were bisected through surgical wounds, and one-half was studied using standard light microscopy. The other one-half specimens were blocked and processed for transmission electron microscopy. The results of the evaluations were as follows: 1. Full thickness sclerectomy wounds healed by ingrowth of fibroblasts from subconjunctival and episcleral tissue, but trabeculectomy wounds showed no communication with extrascleral tissue. 2. Quantitatively, proliferation of fibroblasts were more exuberant in the sclerectomied eyes compared to trabeculectomies. 3. There was no evidence of significant difference between albino and pigmented rabbits. 4. The granulation tissue composed mainly of fibroblasts resembles smooth muscle cell(myofibroblasts) and extracellular collagen constituents. Ultrastructurally, these cells showed massive bundles of intracytoplasmic microfilaments with scattered dense bodies, wrinkled and folded nuclei indicative cellular contraction, and abundant cisternae of rough endoplasmic reticulum informative activity of cellular proliferation. From those results, author conclude that smooth muscle antagonists may inhibit the proliferation of myofibroblasts after glaucoma filtering operations.
Actin Cytoskeleton ; Anesthesia, General ; Cell Proliferation ; Collagen ; Connective Tissue ; Endoplasmic Reticulum, Rough ; Fibroblasts ; Glaucoma ; Granulation Tissue ; Microscopy ; Microscopy, Electron, Transmission ; Muscle, Smooth ; Myofibroblasts ; Rabbits ; Sodium ; Thiopental ; Trabeculectomy ; Wounds and Injuries*

The development and differentiation of cells in the spinal ganglion were studied by electron microscopy in human fetuses ranging from 12 mm to 260 mm crown rump length. At 12 mm embryo the primitive neuroblasts which had a single process, contained a large numbers of free ribosome and mitochondria but very little rough endoplasmic reticulum. At 30 mm fetus, the primitive spinal ganglion consisted of bipolar neuroblasts, satellite cells and undifferentiated cells. Spindle-shaped bipolar neuroblasts formed spinal ganglion of loosely grouped cells at 50 mm fetus. Two neuroblast cell types, a small cell contained large clumps of rough endoplasmic reticulum at periphery, could be distinguished. At 80 mm fetus, the spinal ganglion constituted of bipolar neuroblast with apparently random distribution of small and large neurons with processes, together with satellite cells and blood vessels. The presences of a large numbers of neurotubules in the Golgi-central region were one of the first sign of further maturation of the neuroblast. During next prenatal stage from 120 mm on fetus, the ganglion cells were large and contained much rough endoplasmic reticulum, neurotubules and extensive Golgi complex. A large number of neuroblasts became transformed into unipolar cells from 180 mm to 260 mm feuts. Nissl bodies appeared during this stage. The ganglion-satellite cell boundary became complicated with increasing age, then enlarging in parallel with the increase in volume of the nerve cell. During next prenatal stage up to 180 mm fetus, the unipolar ganglion cell increased in number and size, and the cytoplsm contained all intracytoplasmic structures which were also found in mature spinal ganglion except for large pigment granules.
Blood Vessels ; Crown-Rump Length ; Embryonic Structures ; Endoplasmic Reticulum, Rough ; Fetus* ; Ganglia, Spinal* ; Ganglion Cysts ; Golgi Apparatus ; Humans* ; Microscopy, Electron ; Mitochondria ; Neurons ; Nissl Bodies ; Ribosomes