Modification of the Na,K-pump of glial cells within cobalt-induced epileptogenic cortex of rat

The characteristics of a glial Na⁺,K⁺-pump dependent on extracellular K⁺ within epileptogenic cortex were studied electrophysiologically, biochemically and histochemically in vitro using slices from cobalt-induced epileptogenic cortex of rat. When the extracellular K⁺ concentration ([K⁺]_o) was varied between 4 and 40 mM, the mean slope of membrane potential plotted against [K⁺]_o was about 57 mV in glia from the normal cortex (tissue A) and about 44 mV in glia from the epileptogenic cortex (tissue B); whereas no significant difference in the resting membrane potential of these tissues was observed. In glia from tissue B, a marked transient hyperpolarization above control level was caused by replacement of elevated [K⁺]_o with the normal medium. Ouabain abolished these phenomena observed in glia from tissue B, but had no effect on the membrane potential during normal [K⁺]_o. Reduction of extracellular Na⁺, Ca²⁺ and Cl⁻ did not significantly affect the membrane potential of glia from either tissue. In tissue A, the cells marked by intracellular injection of horseradish peroxidase after intracellular recording were protoplasmic astrocytes; in tissue B, fibrous astrocytes with abnormal processes predominated. K⁺-dependent stimulation of Na⁺,K⁺-ATPase activity of the astrocyte-enriched fraction and its membrane preparation from tissue B was much larger than that from tissue A. A certain amount of the reaction product of K⁺-pNPPase activity was seen on glial plasma membrane within tissue B but not on that from tissue A. The above findings suggest that a glial Na⁺,K⁺-pump within actively firing epileptogenic cortex may be modified to increase in its activity.     

Ultrastructure of lentigo maligna in the oral cavity

A rare case of lentigo maligna in the oral cavity was investigated by light and electron microscopy. Cutaneous lentigo malignas often develop to malignant melanomas. However, the electron microscopic examination revealed that even though there were a large number of melanosomes, most of them were late stage and had membrane structure, and positive staining with HMB-45 was not recognized. From our findings, it is difficult to conclude that oral lentigo malignas develop malignant melanomas, and thus further studies are needed.     

Oral squamous cell carcinoma: electron microscopic and immunohistochemical characteristics

Oral squamous cell carcinoma is the most common oral malignancy, and we performed electron microscopic and immunohistochemical investigation of the tumor. In patients with cervical metastasis, microvilli were developed and a small number of desmosomes were found, regardless of the width of the intercellular spaces. In patients without the metastasis, few microvilli were found in relatively wide intercellular spaces, or numerous microvilli were found in narrow intercellular spaces, and a large number of desmosomes were shown. However, these findings were different from those of tumors that had received radiotherapy, in which numerous microvilli and a small number of desmosomes were found in the nonmetastatic cases. Transferrin receptor, which is a marker of cell proliferation, was localized on the cell membrane, especially in microvilli. Ultrastructural similarity between the primary tumor and the metastatic tumor was recognized, however, the features of microvilli, desmosomes, and the intercellular spaces differed between them in most cases. It is suggested that microvilli might be related to the metastasis of oral squamous cell carcinoma cells. Immunohistochemically, the protein expression of p53 and pRb2/p130 was related to the clinical course of the patients with oral squamous cell carcinoma; the mechanism of the synthesis of these proteins should be investigated in order to understand the biological behavior of the tumor.     

Ultrastructure of oral sarcoma

Sarcoma of the oral region is extremely rare and ultrastructural studies of the tumor are limited in number. We collected oral sarcomas, such as fibrosarcoma, malignant fibrous histiocytoma, liposarcoma, leiomyosarcoma, rhabdomyosarcoma, angiosarcoma, alveolar soft-part sarcoma, solitary plasmacytoma, and osteosarcoma, and performed ultrastructural studies of these tumors. The value of these studies for an understanding of the biological behavior of the tumors was then investigated. In these studies, electron microscopic examinations of oral sarcoma were of assistance in our attempt to establish correct diagnosis and histogenesis. Data from the studies of oral sarcoma by light microscopy, electron microscopy, and immunohistochemistry should be accumulated.     

Scanning electron microscopic study of the human auditory ossicles

The human mallei, incudes and stapedes from 34 cadavers were examined using scanning electron microscope (SEM) to compare the bone surface type among different regions of auditory ossicles for males and females. On the malleus of both males and females, almost all of the surfaces showed a smooth fibrous appearance, characteristic of resting surface. Limited bone-forming or resorbing surfaces were identified on the malleus. As compared with the malleus, the percentage area of the resorbing surface and the vascular canal openings were higher on the incus and stapes, especially on the long process (Crus longum) of the incus and the neck of the stapes for both males and females. The percentage area occupied by the resorbing surface of the long process of the incus and the neck of the stapes correlated with that of the vascular canal openings. We consider that the malleus maintained the stable condition, while the long process of the incus and the neck of the stapes demonstrated marked bone resorption. We suppose that the bone erosion may be related to the vascularization in these regions. Though the percentage area of the resorbing surface and the vascular canal openings had the tendency to be high in females, we did not find any significant differences between the males and females. There was no significant correlation between the age and the area of resorbing surface or vascular canal openings.     

Analysis of proteins in urinary tract stones and urine of urolithic patients

In order to investigate the mechanism of urinary tract stone formation, we analyzed protein components in urine and the stone. Urinary proteins of healthy subjects and urolithic patients as well as protein components urinary tract stone of the urolithic patients were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Electrophoretic patterns of urinary proteins of the patients differed from those of healthy subjects after separating protein patterns into those larger than 66kDa or smaller than 30kDa. Protein constituents of urinary tract stone were mainly separated into 18 bands ranging from 26.8 to 143 kDa. Major bands among these 18 bands differed among stones from different patients. On western blotting, the developed intensities of Tamm-Horsfall protein (THP) were fainter than those of healthy subjects. Whereas intensities of albumin (ALB) were stronger than those of healthy subjects. Moreover, blotting patterns of THP of the patients on non-reducing SDS-PAGE were obviously broad. Thus, we suggest that analysis of fractionated urinary proteins or protein components of urinary tract stone may provide a tool for monitoring the prognosis or relapse in the patients.     

Identification of Dish-Based Dietary Patterns for Breakfast,Lunch, and Dinner and Their Diet Quality in Japanese Adults

We identified dish-based dietary patterns for breakfast, lunch, and dinner and assessed the diet quality of each pattern. Dietary data were obtained from 392 Japanese adults aged 20–69 years in 2013, using a 4 d dietary record. K-means cluster analysis was conducted based on the amount of each dish group, separately for breakfasts (n = 1462), lunches (n = 1504), and dinners (n = 1500). The diet quality of each dietary pattern was assessed using the Healthy Eating Index 2015 (HEI-2015) and Nutrient-Rich Food Index 9.3 (NRF9.3). The extracted dietary patterns were as follows: ‘bread-based’ and ‘rice-based’ for breakfast; ‘bread’, ‘rice-based’, ‘ramen’, ‘udon/soba’, and ‘sushi/rice bowl dishes’ for lunch; and ‘miscellaneous’, ‘meat dish and beer’, and ‘hot pot dishes’ for dinner. For breakfast, the HEI-2015 and NRF9.3 total scores were higher in the ‘rice-based’ pattern than the ‘bread-based’ pattern. For lunch, the HEI-2015 and NRF9.3 total scores were relatively high in the ‘rice-based’ pattern and low in the ‘ramen’ pattern. For dinner, the HEI-2015 total score was the highest in the ‘meat dish and beer’ pattern, and the NRF9.3 total score was higher in the ‘hot pot dishes’ than the ‘miscellaneous’ pattern. These results suggested that breakfast, lunch, and dinner have distinctive dietary patterns with different diet qualities.     

Effects of dorzolamide hydrochloride on ocular tissues.

We studied the intraocular pharmacokinetics of dorzolamide hydrochloride eye drops and the effect of dorzolamide on carbonic anhydrase activity and localization in ocular tissues. Carbonic anhydrase activity was detected in normal ocular tissues. The activity was inhibited in corneal endothelial cells, the ciliary body, lens epithelial cells, or the retina 1 to 8 hours after instillation of dorzolamide eye drops. In lens epithelial cells and the retina, the enzyme activity had not recovered even 10 hours after instillation of the drug. Immunostaining did not reveal any differences between the group administered dorzolamide eye drops and the control group administered a physiologically balanced solution. Time-related changes in dorzolamide concentrations in ocular tissues were measured by high-performance liquid chromatography (HPLC). In the cornea, anterior aqueous, iris, ciliary body and retina, drug concentrations increased 15 minutes after the instillation and peaked within 1 hour. These results suggest that dorzolamide immediately suppresses carbonic anhydrase activity in ocular tissues, and is rapidly distributed among the tissues of the eye when administered as eye drops.