The authors have demonstrated the effect of sodium selenite on the hepatotoxicity due to carbon tetrachloride, by observing the distribution and disaggregation of the pyroninophilic granules in the hepatic cell of the mature male albino mice. Each experimental mouse of the selenite and the selenite plus carbon tetrachloride groups was given a single dose of 4 ug. of sodium selenite per kilogram of body weight and that of the control and the carbon tetrachloride groups was given 0.1 ml. of distilled water alone. Six hours after the first administration of distilled water or sodium selenite, the experimental mice of the carbon tetrachloride and the selenite plus carbon tetrachloride groups were given a single dose of l.0 ml. of carbon tetrachloride per kilogram of body weight and those of the selenite groups were given 0.l ml. of paraffin oil alone. Following the 1ast administration of carbon tetrachloride or paraffin oil, the mice were sacrificed by bleeding (cutting the common carotid artery) at the intervals of 2,3,4,6,8, and 12 hours respectively. Histochemical preparations were stained by the methyl-green and pyronin method and oil red 0 method. The hepatotoxicity due to the administration of carbon tetrachoride was evident in the hepatic cells; the pyroninophilic granlues were partly reduced in volume in the hepatic cells of the centrilobular and the intermediate zones as early as the 3 hour-period, and markedly reduced or disappeared in the centrilobular and some part of the intermediate zones associated with hydropic degeneration as well as in the 6 hour-period. Thereafter marked reduction or dissolution of the pyroninophilic granules was found and extended as the periportal zone at the 12 hour-period. However, the pyroninophilic granules in the hepatic cells of selenite plus carbon tetrachbride group showed no significant changes in the hepatic cells of these zones, compared to the histochemical feature of the granules in the hepatic cells of the control and the selenite groups. Consequently it is suggested that the lipid peroxidative decomposition of the microsomal membranes, which is induced with carbon tetrachloride, would be prevented by a previous administration of sodium selenite.
Animal ; Carbon Tetrachloride Poisoning*/pathology ; Cell Nucleus/drug effects ; Cytoplasm/drug effects ; Cytoplasmic Granules ; Lipids ; Liver/drug effects* ; Liver/pathology ; Male ; Mice ; Selenium/pharmacology* ; Vacuoles/drug effects

OBJECTIVETo investigate whether tri-ortho-cresyl phosphate (TOCP) and organophosphate compound that could produce organophosphate-induced delayed neuropathy (OPIDN) in hen and other sensitive species, directly affect oligodendrocytes, the myelin-forming cell of the central nervous system.

METHODSThis was achieved by a combination of measurements of cell viability (MTT) cell pathological observation in the presence and absence of the compound cultured hen brain oligodendrocytes were prepared and treated with various concentrations of TOCP.

RESULTSIn a time-course experiment TOCP showed a cytotoxic effect to oligodendrocytes and led to the oligodendrocyte processes disintegrated and membranous blebs, cytoplasmic vacuolation following exposure time of 24 h or longer, it showed a dose-depended and time-depended manner cytotoxic effect to oligodendrocytes at dose levels of 0.5 approximately 1.5 microg/ml (1.35 approximately 4.05 mol/L) concentrations of TOCP for 24 - 72 h exposure. MTT experiment indicated that TOCP inhibited cell viability by dose-depended manner at dose levels of 0.5 approximately 1.5 microg/ml (1.35 approximately 4.05 mol/L) concentrations of TOCP for an 24 h exposure.

CONCLUSIONSTOCP is cytotoxic to oligodendrocytes and leads to the oligodendrocyte processes disintegrated and membranous blebs, vacuolar degeneration, which suggests that this oligodendrocyte degeneration may involve in the pathogenesis mechanism for OPIDN.

Animals ; Cell Survival ; Cells, Cultured ; Cerebral Cortex ; pathology ; Chickens ; Dose-Response Relationship, Drug ; Oligodendroglia ; drug effects ; pathology ; Tritolyl Phosphates ; toxicity ; Vacuoles ; drug effects ; pathology

We describe a 59-year-old female with severe anticonvulsant hypersensitivity syndrome (AHS) associated with Epstein- Barr virus (EBV) infection. The causative drug was speculated to be carbamazepine. Recurrent EBV infection was demonstrated by the presence of anti-EBV early antigen IgM antibodies and anti-EBV nuclear antigen IgG antibodies. To our knowledge, only one case of drug hypersensitivity syndrome (DHS) associated with EBV has been reported in the English- language literature. Our case is the second report of EBV-associated DHS, which suggests that EBV infection may contribute to the pathogenesis of AHS in a few patients.
Virus Activation/*physiology ; Vacuoles/pathology ; Middle Aged ; Humans ; Herpesvirus 4, Human/drug effects/pathogenicity ; Female ; Erythema/etiology/virology ; Epstein-Barr Virus Infections/*physiopathology ; *Drug Hypersensitivity ; Anticonvulsants/*adverse effects

Amyloid beta (Abeta) neurotoxicity is believed to play a critical role in the pathogenesis of Alzheimer's disease (AD) mainly because of its deposition in AD brain and its neuronal toxicity. However, there have been discrepancies in Abeta-induced cytotoxicity studies, depending on the assay methods. Comparative analysis of Abeta42-induced in vitro cytotoxicity might be useful to elucidate the etiological role of Abeta in the pathogenesis of AD. In this study, MTT, CCK-8, calcein-AM/EthD-1 assays as well as thorough microscopic examinations were comparatively performed after Abeta42 treatment in a neuronal precursor cells (NT2) and a somatic cells (EcR293). Extensive formation of vacuoles was observed at the very early stage of Abeta42 treatment in both cells. Early observation of Abeta42 toxicity as seen in vacuole formation was also shown in MTT assay, but not in CCK-8 and calcein-AM/EthD-1 assays. In addition, Abeta42 treatment dramatically accelerated MTT formazan exocytosis, implying its effect on the extensive formation of cytoplasmic vacuoles. Abeta42 seems to cause indirect inhibition on the intracellular MTT reduction as well as vacuole formation and exocytosis enhancement. Following the acute cellular dysfunction induced by Abeta42, the prolonged treatment of micromolar concentration of Abeta42 resulted in slight inhibition on redox and esterase activity. The early Abeta42-induced vacuolated morphology and later chronic cytotoxic effect in neuronal cell might be linked to the chronic neurodegeneration caused by the accumulation of Abeta42 in AD patients' brain.
Amyloid beta-Protein/*toxicity ; Animals ; Cell Death/drug effects ; Cell Line ; Dose-Response Relationship, Drug ; Exocytosis/*drug effects ; Formazans ; Neurons/*drug effects/metabolism/*pathology ; Peptide Fragments/*toxicity ; Research Support, Non-U.S. Gov't ; Tetrazolium Salts ; Time Factors ; Vacuoles/*drug effects

PURPOSE: To evaluate the anti-proliferative effect of mitomycin C (MMC) on human corneal keratocyte, and to investigate the cellular morphology of keratocyte according to the concentration and exposure time in vitro. METHODS: Human corneal keratocytes using endothelium-free explant method were exposed to 0.005%, 0.01%, and 0.05% concentration of MMC for 3, 5, and 10 minutes. MTT based colorimetric assay was performed to assess the inhibition of cellular proliferation, and cellular morphology was evaluated by inverted phase-contrast light microscope and electron microscope. RESULTS: Use of higher concentration MMC and prolongation of exposure time resulted in greater inhibitory effect on cellular proliferation. When exposed to 0.005% MMC for 3, 5 and 10 minutes, the survival rate of keratocyte was 100%, 95.7% and 74.0% respectively. At 0.01% MMC, the survival rate was 98.6%, 92.9%, and 66.9%. At 0.05% MMC, it was 74.0%, 73.4%, and 38.8%. Exposure to the highest concentration (0.05%) among the 3 preparations for 3 or 5 minutes showed significant inhibition of keratocyte proliferation (p<0.05), and when exposed for 10 minutes, all 3 preparations showed significant inhibition of keratocyte proliferation (p<0.05). Inverted phase-contrast light microscopy showed that human corneal keratocytes lost their adherence to the bottom of the dish and assumed round and swollen shape rather than spindle shape when exposed to higher concentration of MMC for a prolonged time. The damaged keratocytes showed the degenerative changes like cellular membrane disruption, disappearance of microvilli, enlargement of rough surfaced endoplasmic reticulum and mitochondria, and vacuole formation by electronic microscope. CONCLUSIONS: When MMC is applied to inhibit the proliferation of keratocytes involved in corneal wound healing, it seems to be a valuable application at least 0.05% concentration for 3 minutes. Further studies should be followed for the biological effect of MMC including drug toxicity associated with human corneal tissue in vivo.
Cell Proliferation ; Corneal Keratocytes* ; Drug-Related Side Effects and Adverse Reactions ; Endoplasmic Reticulum ; Humans* ; Membranes ; Microscopy ; Microvilli ; Mitochondria ; Mitomycin* ; Survival Rate ; Vacuoles ; Wound Healing

AIM: To investigate the histological and ultrastructural changes observed in pan masala intoxicated mammalian testes under the effect of cardamom. METHODS: Male Swiss mice were given pan masala orally at a dose of 2% of the feed and cardamom at a dose of 0.2% of the feed. They were divided into three groups, control (Group I), pan masala-treated (Group II), and a combination of pan masala and cardamom-treated group (Group III). Histologically, the testes of Group II mice displayed degeneration of tubular epithelium, disruption of spermatogenesis, and a marked reduction in germ cells. RESULTS: When cardamom was given, damage was less with fewer distorted cells and also improvement with normal tubules and spermatid differentiation in Group III. Ultrastructurally, pan masala-treated testes showed cytoplasmic vacuolation, shrinkage and pyknotic nuclei of spermatogonia, and abnormal acrosomal granules. CONCLUSION When cardamom was given, the amelioration process was more evident showing a comparable morphology with control.
Animals ; Areca ; adverse effects ; Elettaria ; Male ; Mice ; Phytotherapy ; Plant Extracts ; pharmacology ; therapeutic use ; Spermatogenesis ; drug effects ; Spermatozoa ; drug effects ; Testicular Diseases ; chemically induced ; drug therapy ; pathology ; Testis ; drug effects ; pathology ; ultrastructure ; Tobacco ; adverse effects ; Tobacco, Smokeless ; adverse effects ; Vacuoles ; drug effects

In this study, the effects of mono(2-ethylhexyl) phthalate (MEHP), one of metabolites of di(2-ethylhexyl) phthalate, on immature Shiba goat testes in vitro were examined. The testes of 2-month-old Shiba goats were cut into smaller pieces, and seeded in medium. At 1, 3, 6 and 9 hr after administration of MEHP at various concentrations (0, 100 nmol ml-1, 1 nmol ml-1, and 1 x 10-3 nmol ml-1, respectively), the specimens were obtained for light and transmission electron microscopic observations. As a result, at 1 hr after exposure to MEHP, the vacuolization and nuclear membrane rupture appeared in Sertoli cells. Such alterations tended to gradually increase in number in timeand dose-dependent manners. Moreover, by MEHP treatment, apoptotic spermatogenic cells (characterized with chromatin condensation, cytoplasm shrinkage without membrane rupture, still functioning cell organelles, and packed cell contents in membrane-bounded bodies), apoptotic Sertoli cells (characterized with nuclear membrane lysis, nuclear condensation), necrotic spermatogenic cells (characterized with swollen and ruptured mitochondria, plasma membrane lysis, spilt cell contents, and chromatin clumps), and necrotic Sertoli cells (characterized with marginated chromatins along the nuclear membrane, ruptured vesicles within the MNB, some swollen and ruptured cell organelles, e.g. mitochondria) could be identified. Conclusively, ultrastructurally the treatment with MEHP at low concentration tends to lead spermatogenic and Sertoli cells to apoptosis, whereas that at high concentration tends to lead spermatogenic and Sertoli cells to necrosis. Thus, the testicular tissue culture is advantageous for screening testicular toxicity of chemicals.
Animals ; Apoptosis/drug effects/physiology ; Diethylhexyl Phthalate/*analogs&derivatives/*toxicity ; Goat Diseases/*chemically induced/pathology ; Goats ; Male ; Microscopy, Electron, Transmission/veterinary ; Necrosis ; Sertoli Cells/ultrastructure ; Spermatozoa/drug effects/pathology/ultrastructure ; Testicular Diseases/*chemically induced/pathology ; Testis/*drug effects/metabolism/pathology/ultrastructure ; Vacuoles/physiology/ultrastructure

BACKGROUNDThe loss of cardiac myocytes is one of the mechanisms involved in acute myocardial infarction (AMI)-related heart failure. Autophagy is a common biological process in eukaryote cells. The relationship between cardiac myocyte loss and autophagy after AMI is still unclear. Carvedilol, a non-selective alpha1- and beta-receptor blocker, also suppresses cardiac myocyte necrosis and apoptosis induced by ischemia. However, the association between the therapeutic effects of carvedilol and autophagy is still not well understood. The aim of the present study was to establish a rat model of AMI and observe changes in autophagy in different zones of the myocardium and the effects of carvedilol on autophagy in AMI rats.

METHODSThe animals were randomly assigned to a sham group, an AMI group, a chloroquine intervention group and a carvedilol group. The AMI rat model was established by ligating the left anterior descending coronary artery. The hearts were harvested at 40 minutes, 2 hours, 24 hours and 2 weeks after ligation in the AMI group, at 40 minutes in the chloroquine intervention group and at 2 weeks in other groups. Presence of autophagic vacuoles (AV) in the myocytes was observed by electron microscopy. The expression of autophagy-, anti-apoptotic- and apoptotic-related proteins, MAPLC-3, Beclin-1, Bcl-xl and Bax, were detected by immunohistochemical staining and Western blotting.

RESULTSAVs were not observed in necrotic regions of the myocardium 40 minutes after ligation of the coronary artery. A large number of AVs were found in the region bordering the infarction. Compared with the infarction region and the normal region, the formation of AV was significantly increased in the region bordering the infarction (P < 0.05). The expression of autophagy- and anti-apoptotic-related proteins was significantly increased in the region bordering the infarction. Meanwhile, the expression of apoptotic-related proteins was significantly increased in the infarction region. In the chloroquine intervention group, a large number of initiated AVs (AVis) were found in the necrotic myocardial region. At 2 weeks after AMI, AVs were frequently observed in myocardial cells in the AMI group, the carvedilol group and the sham group, and the number of AVs was significantly increased in the carvedilol group compared with both the AMI group and the sham group (P < 0.05). The expression of autophagy- and anti-apoptotic-related proteins was significantly increased in the carvedilol group compared with that in the AMI group, and the positive expression located in the infarction region and the region bordering the infarction.

CONCLUSIONSAMI induces the formation of AV in the myocardium. The expression of anti-apoptosis-related proteins increases in response to upregulation of autophagy. Carvedilol increases the formation of AVs and upregulates autophagy and anti-apoptosis of the cardiac myocytes after AMI.

Adrenergic beta-Antagonists ; pharmacology ; Animals ; Apoptosis ; Apoptosis Regulatory Proteins ; analysis ; Autophagy ; drug effects ; Beclin-1 ; Carbazoles ; pharmacology ; therapeutic use ; Immunohistochemistry ; Male ; Microscopy, Electron, Transmission ; Myocardial Infarction ; drug therapy ; pathology ; Myocardium ; ultrastructure ; Propanolamines ; pharmacology ; therapeutic use ; Rats ; Rats, Wistar ; Vacuoles ; drug effects