OBJECTIVE: The purpose of this study was to evaluate the effect of Cytochalasin B (CCB) on the cytoskeletal stability of mouse oocyte frozen by vitrification. METHODS: Mouse oocytes retrieved from cycle stimulated by PMSG and hCG were treated by CCB and then vitrified in EFS-30. These oocytes were placed onto an EM grid and submerged immediately in liquid nitrogen. Thawing of the oocytes was carried out at room temperature for 5 seconds, then the EM grid was placed into 0.75 M, 0.5 M and 0.25 M sucrose at 37degress C for 3 minutes, each. These oocytes were fixed in 4% formaldehyde for an hour and then washed in PPB for 15 minutes 3 times, then incubated in PPB containing anti-tubulin monoclonal antibody at 4degress C overnight. And then, the oocytes were incubated with FITC-conjugated anti-mouse IgG and propidium iodide (PI) for 45 minutes. Pattern of microtubules and microfilaments of oocytes were evaluated with a confocal microscope. RESULTS: The rate of oocytes containing normal microtubules and microfilaments was significantly decreased after vitrification. The rate of oocyte containing normal microtubules in CCB treated group was higher than those in non-treated group (53.7% vs. 48.9%), but the difference was not significant. The rate of oocyte containing normal microfilaments in CCB treated group was significantly higher than those in non-treated group (64.5% vs. 38.3%, p<0.05).CONCLUSION: Microfilaments stability could be improved by CCB treatment prior to vitrification. It is suggested that CCB treatment prior to vitrification improve stability of cytoskeleton and then increase success rate in IVF-ET program using vitrification and thawing oocyte.
Actin Cytoskeleton ; Animals ; Cytochalasin B* ; Cytoskeleton ; Formaldehyde ; Immunoglobulin G ; Mice* ; Microtubules ; Nitrogen ; Oocytes* ; Propidium ; Sucrose ; Vitrification*

OBJECTIVE: The purpose of this study was to evaluate the effect of Cytochalasin B (CCB) on the cytoskeletal stability of mouse oocyte frozen by vitrification. METHODS: Mouse oocytes retrieved from cycle stimulated by PMSG and hCG were treated by CCB and then vitrified in EFS-30. These oocytes were placed onto an EM grid and submerged immediately in liquid nitrogen. Thawing of the oocytes was carried out at room temperature for 5 seconds, then the EM grid was placed into 0.75 M, 0.5 M and 0.25 M sucrose at 37degress C for 3 minutes, each. These oocytes were fixed in 4% formaldehyde for an hour and then washed in PPB for 15 minutes 3 times, then incubated in PPB containing anti-tubulin monoclonal antibody at 4degress C overnight. And then, the oocytes were incubated with FITC-conjugated anti-mouse IgG and propidium iodide (PI) for 45 minutes. Pattern of microtubules and microfilaments of oocytes were evaluated with a confocal microscope. RESULTS: The rate of oocytes containing normal microtubules and microfilaments was significantly decreased after vitrification. The rate of oocyte containing normal microtubules in CCB treated group was higher than those in non-treated group (53.7% vs. 48.9%), but the difference was not significant. The rate of oocyte containing normal microfilaments in CCB treated group was significantly higher than those in non-treated group (64.5% vs. 38.3%, p<0.05).CONCLUSION: Microfilaments stability could be improved by CCB treatment prior to vitrification. It is suggested that CCB treatment prior to vitrification improve stability of cytoskeleton and then increase success rate in IVF-ET program using vitrification and thawing oocyte.
Actin Cytoskeleton ; Animals ; Cytochalasin B* ; Cytoskeleton ; Formaldehyde ; Immunoglobulin G ; Mice* ; Microtubules ; Nitrogen ; Oocytes* ; Propidium ; Sucrose ; Vitrification*

Brainstem auditory response abnormalities directly reflect disturbance of neural function rather than the underlying anatomic cause of that disturbance. The test has advantage in detecting lesions that alter electrophysiology but do not produce detectable alterations of radiodensity, displace surrounding structures or change vascular supply and permeability. A sequence of seven low-amplitude potentials that occur in the initial 10 msec following click signals can be recorded from scalp electrodes in 44 patient with focal brain lesions using computer averaging techniques. The potentials, termed auditory brainstem responses, are thought to be the far-field reflection of electrical events originating in the auditory pathway during its course through the brainstem. We have studied auditory brainstem evoked potential responses in a variety of focal brain lesions and found them to be of assistance in evaluating the localization of pontomedullary, pons, midbrain, thalamus, subcortical and functional recovery. 1) Distortion of early components (type I) was occured in the brainstem lesions. 2) Distortion of late components (type II) was developed in the diencephalon or subcortical lesions. 3) Distortion of all components (type III) was developed in the brainstem and diffuse brain contusions. 4) Serial recordings provided information about the evolution of brain stem lesions and good functional recovery marker.
Auditory Pathways ; Brain Injuries ; Brain Stem* ; Brain* ; Diencephalon ; Electrodes ; Electrophysiology ; Evoked Potentials* ; Evoked Potentials, Auditory, Brain Stem ; Humans ; Mesencephalon ; Permeability ; Pons ; Scalp ; Thalamus

Brainstem auditory response abnormalities directly reflect disturbance of neural function rather than the underlying anatomic cause of that disturbance. The test has advantage in detecting lesions that alter electrophysiology but do not produce detectable alterations of radiodensity, displace surrounding structures or change vascular supply and permeability. A sequence of seven low-amplitude potentials that occur in the initial 10 msec following click signals can be recorded from scalp electrodes in 44 patient with focal brain lesions using computer averaging techniques. The potentials, termed auditory brainstem responses, are thought to be the far-field reflection of electrical events originating in the auditory pathway during its course through the brainstem. We have studied auditory brainstem evoked potential responses in a variety of focal brain lesions and found them to be of assistance in evaluating the localization of pontomedullary, pons, midbrain, thalamus, subcortical and functional recovery. 1) Distortion of early components (type I) was occured in the brainstem lesions. 2) Distortion of late components (type II) was developed in the diencephalon or subcortical lesions. 3) Distortion of all components (type III) was developed in the brainstem and diffuse brain contusions. 4) Serial recordings provided information about the evolution of brain stem lesions and good functional recovery marker.
Auditory Pathways ; Brain Injuries ; Brain Stem* ; Brain* ; Diencephalon ; Electrodes ; Electrophysiology ; Evoked Potentials* ; Evoked Potentials, Auditory, Brain Stem ; Humans ; Mesencephalon ; Permeability ; Pons ; Scalp ; Thalamus

This experiment was performed to define the paricipation of a discrete hypothalamic neural structure in the genesis of pulmonary edema and the effect of alpha adrenergic blockade. Fifty adult cats weighing 2.5 to 4.0 Kg, were used in this study. The components of the pathophysiological systemic changes, lung weight, and histopathological changes of lung and hypothalamus were studied in groups of animals when intracranial pressure(ICP) was raised to 200 mmH2O of 300 mmH2O for 2 hours by intraventricular infusion with normal saline. The animals were divided into 5 groups : The normal control group was comprised in 10 normal cats. Control and phentolamine treated animal groups which had an elevated ICP of up 200 mmH2O consisted of 10 cats each. Control and phentolamine treated animal groups which had an elevated ICP of up to 300 mmH2O consisted of 10 cats each. The results obtained were as follows : 1) In the animal groups of elevated ICP to 200 mmH2O or 300 mmH2O, there were hemodynamic systemic changes which were neurogenically mediated and caused an immediate elevation in blood pressure of 30 mmHg to 60 mmHg. The hemodynamic data of the animals that had an elevated ICP of up to 300 mmH2O were significantly more deviated from normal control values than the 200 mmH2O ICP groups. The hemodynamic responses of the phentolamine treated animal with elevated ICP of up to 200 and 300 mmH2O were less deviated from normal control values. 2) The lung weights of the animals with an elevated ICP of up to 200 and 300 mmH2O were significantly heavier than the normal control value(p<0.05) and the lung weights of the animals with an elevated ICP of 300 mmH2O were significantly heavier than those with an ICP of 200 mmH2O(p<0.01). The lung weights of the phentolamine treated animal groups were significantly lighter than the control group but showed little increase in the lung weight when compared to the normal value. 3) By controlling the elevated ICP above 200 mmH2O in the experimental animals we have confirmed gross and microscopic appearances of hemorrhagic pulmonary edema. Histopathological changes of the phentolamine treated animals were significantly less sever than in the control groups. 4) By elevating ICP above 200 mmH2O in the experimental animals, we have confirmed discrete bilateral hemorrhagic spots of the anterior hypothalamus, preoptic region induced by increased intracranial pressure. Histopathological changes of the phentolamine treated animals with the elevated ICP were significantly less severe than of the control groups. 5) This experimental model may define the specific particification of the hypothalamus in the pathophysiological pathogenesis of neurogenic pulmonary edema. These results suggest that the lungs are directly affected by the intense sympathetic discharge evoked by release phenomenon from the sympathoinhibitory influence of the hypothalamus, and pulmonary edema was effectively eliminated by alpha adrenergic blockade.
Adult ; Animals ; Blood Pressure ; Cats* ; Edema* ; Hemodynamics ; Humans ; Hypothalamus ; Hypothalamus, Anterior ; Infusions, Intraventricular ; Intracranial Pressure ; Lung* ; Models, Theoretical ; Phentolamine* ; Pulmonary Edema ; Reference Values ; Weights and Measures