Several technical procedures were described for estimation of epididymal sperm concentration until today. In all of the described techniques, epididymal material was drawn into a micro-pipette before counting procedure. We aimed to proceed this stage without using any pipette because of the difficulty in aspirating very viscous epididymal material into a micro-pipette. For measuring epididymal material we used a 2 μl reservoir which we named as a micro-spoon. It was filled with the epididymal material and layered in a tube with Ringer's solution of 2 ml containing sodium hyaluronate. Then, it was mixed in order to obtain 1/1000 diluted suspension. Sperm cells in this suspension were counted on a Makler chamber under a light microscope. The procedure was easy and time saving, compared to the other techniques.
Autologous iris pigment epithelial cell transplantation was performed on patients with exudative age-related macular degeneration (AMD). Autologous IPE cell culture was performed using autologous serum after iridectomy in 7 patients with AMD. The cell suspensions (2∼20×104 cells) were transplanted into the submacular lesion of individuals after removal of neovascular membranes. Subsequent ophthalmological examinations, including best corrected visual acuity and fluorescein or indocyanine green angiography, were performed. In addition, 15 patients with AMD, who underwent removal of neovascular membrane without transplantation, were evaluated as non randomized controls. Varying degrees of atrophy or defects of choriocapillaris and retinal pigment epithelium were observed in all of the patients. No cystoid macular edema or fluorescein leakage was observed after treatment, but window defects were present. No patient had decreased visual acuity. One treated patient developed mild subretinal fibrosis and an other patient developed mild preretinal fibrosis, however no difference was significant when compared with the control. In coclusion, the treatment resulted in no significant improvement in macular function, as compared with the control; however, no rejection or deterioration in visual acuity occurred up to the 13 month follow up.
Tissue Doppler imaging is a new ultrasound technology that derives measurements of contraction and relaxation velocities directly from the myocardium. However, data on myocardial velocities by using tissue Doppler imaging have not been established in normal children. In 48 normal children, myocardial velocities were measured using tissue Doppler imaging at three different sites (base, middle, and apex) in the left and right ventricles and the interventricular septum. In the left ventricular wall, the peak myocardial velocities during early diastole (peak E), during atrial contraction (peak A), and during systole (peak S) waves decreased gradually between the base and apical sites, whereas the ratio of E to A waves (peak E/A wave ratio) did not change among the 3 segments. Similar findings were obtained from the myocardial velocities in the right ventricle and the interventricular septum. A systolic and diastolic velocity gradient was also observed between the different ventricular walls. Significant correlations of the tissue Doppler parameters with age or heart rate were observed. In the left ventricle, the peak E wave demonstrated a stronger relation with age (r=0.77) than with heart rate (r=−0.65). The peak A wave did not change with age but correlated with heart rate. The peak E/A wave ratio showed a weaker relation with age (r=0.54) than with heart rate (r=0.62). The peak S wave was related to age (r=0.65) and to a lesser extent to heart rate (r=−0.51). Similar relationships of tissue Doppler parameters with age or heart rate were observed for the right ventricle and interventricular septum. The heterogeneous pattern and age- and heart-rate-related changes in normal myocardium demonstrated in this study must be taken into account when attempting to identify altered regional myocardial function with tissue Doppler echocardiography.
We report the peculiar serial electroencephalographic (EEG) findings in a 7-year-old boy with glutaric aciduria type 1 during an episode of acute encephalopathy. The patient developed Reye-like syndrome triggered by cellulitis. Cranial magnetic resonance imaging demonstrated diffuse softening of cerebral hemisphere. The EEG on the day following onset of acute encephalopathy showed suppression burst pattern including continuous 14-15 Hz rhythmic waves at first. Then, periodic synchronous discharge appeared and lasted for about 40 minutes. Periodic synchronous discharge finally disappeared and nearly total electrocerebral silence continued. There have been no reports indicating such a change of EEG in a short period. The serial EEG changes probably reflect the process of electrical death of neurons in cerebral hemispheres.
Protein phosphatase type-1 (PP1), one of the most abundant Ser/Thr protein phosphatases, plays a central role in the regulation of various cell functions. Almost all the PP1 molecules exist as holoenzymes in vivo consisting of a catalytic subunit (PP1C) and a variable regulatory subunit that regulates substrate specificity and/or subcellular localization. In order to clarify fine regulation of PP1, we overexpressed a nuclear inhibitor of PP1 (NIPP-1) in a Flag-tagged form in mammalian cells. The Flag-tagged NIPP-1 was found to be immunoprecipitated with three isoforms of PP1C, namely, PP1α, PP1γ1, and PP1δ with a similar efficiency, suggesting that NIPP-1 makes a complex with the PP1C through the region conserved among the three isoforms. These results suggested that NIPP-1 can be involved in the regulation of various PP1 holoenzymes in vivo.
We investigated the late effects of serotonergic drugs on mouse forced swimming and found that serotonin 1A antagonist NAN-190 hydrobromide (NAN-190) and serotonin 2 agonist R(−)-DOI hydrochloride (DOI) increase the typical anti-depressive behavior climbing 6 hours after the intraperitoneal injection. With the use of ion exchanger, ultra filtration and chemical extraction methods, a substance having remarkable anti-depressive activity on mouse forced swimming was extracted from the serums of the mice treated with the drugs. The substance was strongly suggested to be a glycolipid having a specific sugar chain structure GalNAcα1-3GalNAc in its terminal. In fact, globopentaosylceramide, another glycolipid having the structure in the terminal also showed marked anti-depressive activity on mouse forced swimming, but globotetraosylceramide lacking the sugar chain structure did not. The GalNAcα1-3GalNAc-lipid reactivity in the serum and typical anti-depressive behavior climbing increased corresponding to the doses of NAN-190 and DOI. These findings clearly indicate that the terminal structure GalNAcα1-3GalNAc has the essential role in the anti-depressive activity of globopentaosylceramide, and that serotonin 1A antagonism and serotonin 2 agonism increase the production of an anti-depressive glycolipid having the terminal structure in mouse serum.
A new method which facilitates identification of the internal auditory canal during the middle cranial fossa approach is described. This method uses illumination of the external auditory meatus as a guide to the internal auditory canal. When the external auditory meatus is illuminated by a light source system such that used for a flexible fiberscope, areas of the middle ear air cell system are illuminated and observable through the plate of bone constituting the base of the middle cranial fossa. In the case of ordinary pneumatization of the middle ear system, the attic is the site of the most intense illumination. Knowledge of the exact location of the attic obtained by this new method is extremely useful when performing drilling to expose the internal auditory canal in the middle cranial fossa approach. This is because the internal auditory canal is located medial to the attic, and also because such knowledge is helpful in identifying the two conventional landmarks of this approach, i.e., the arcuate eminence and the greater superficial petrosal nerve, as the locations of these structures are intimately related to the attic. The method described is very simple, timesaving and inexpensive, and is very helpful in detecting the internal auditory canal. Further anatomical study is being undertaken to establish a method of pinpointing the internal auditory canal, starting from the attic which is readily identifiable by use of this method.