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<P>Normal human fibroblasts have a finite proliferative capacity in vitro. Thus, immortalization of human cells is associated with cellular aging. We have established an immortalization-sensitive cell line from fibroblasts of Wilms' tumor patients which have a partial deletion of chromosome 1 1p. This cell line was easily immortalized by introducing SV4OT. By differential hybridization using both SV4OT-introduced crisis cells and young cells, we cloned a gene that was highly expressed in 1 1p-cells at the time of the crisis and named this gene C-1. Nucleotide sequence analysis of C-1 revealed that it contains a helix-loop-helix domain, indicating that it may be a transcription factor. Expression of the C-1 gene was transiently induced early in the G0-to-S phase transition in two normal human (OUMS-24 and HSF-412) and a non-tumorigenic immortal human (OUMS-24F) fibroblast cell lines, while the other immortal SUSM-1 cells highly expressed the C-1 gene in the middle G1 phase. These results suggest that the C-1 gene product may function as a transcription factor related to the cell cycle.</P>

<P>Phospholipid vesicles, also known as liposomes, were examined for their ability to act as a drug carrier to the brain. 9-Amino-1,2,3,4-tetrahydroacridine (THA), a centrally acting acetylcholinesterase inhibitor, was used as a model drug. THA was encapsulated in dehydration-rehydration vesicles (DRV) composed of egg yolk phosphatidylcholine, cholesterol and dipalmitoyl-phosphatidic acid (molar ratio, 10/10/1) and injected into the heart of mice. The toxicity and side effects of THA were reduced by encapsulation in liposomes. The THA concentration in the mouse brain after injection of THA-encapsulated DRV at a dose of 2 mg/kg remained higher than that of free THA at the same dose. Effective concentration of THA in the brain was also prolonged by the use of liposomes, although accumulation of THA in the spleen and kidney was observed. We, therefore, concluded that liposomes are useful as carriers of drugs to the brain.</P>

<P>Four cases of femoral neck fracture following avascular necrosis of the femoral head were studied histologically. All four patients were women who had received steroid therapy, three of them for systemic lupus erythematosus and the other for idiopathic thrombocytopenic purpura. Two types of fracture were found according to the site and the mechanism of fracture. One was at the junction between the necrotic bone and the repairing bone, and it can thus be regarded as a stress fracture. The other type of fracture commenced at the superior portion of the junction between the femoral head and neck, which was weak due to the repair reaction. The fracture line extended to the inferior cortex of the femoral neck, as often occurs in the elderly. In one patient, the femoral neck fracture was the first sign of avascular necrosis of the femoral head. </P>

This study was undertaken to assess postoperative gastric motility and gastric acid secretion, and pre- and postoperative carbohydrate metabolism in patients with esophageal cancer. The gastric motility was compared among 3 different reconstruction routes in 26 patients who were divided into 2 groups according to the duration of postoperative follow-up; group A, 3 months or less; and group B, 18 months or more. The routes used for subtotal resection of the stomach were the posterior mediastinal, retrosternal, and subcutaneous routes. All patients showed positive resting pressure in the esophagus, but peristaltic waves did not reach the gastric tube at dry swallowing in any patients and peristaltic waves appeared after eating pudding only in 1 patient in group B. The resting pressure and gastric emptying time were similar among reconstruction routes, but the incidence and amplitude of metoclopramide (MCP)-induced peristaltic waves were significantly higher in group B than in group A. Furthermore, 24-h intragastric pH monitoring of gastric secretion in a group of 9 patients revealed individual variation in gastric secretion. Some patients showed high acidity soon after operation, suggesting the need for prophylactic treatment for preventing gastric ulcer. Postoperatively, postprandial serum gastrin levels were significantly higher than preoperative levels. In the other group of 11 patients tested, preoperative and postoperative carbohydrate metabolism were not significantly different. Postoperatively, carbohydrate metabolism recovered to preoperative levels after a transient decrease. These results demonstrated that postoperative motility improved over time although no difference was found among the 3 reconstruction routes used.

To elucidate the latent state and reactivation of Epstein-Barr virus (EBV) in non-neoplastic lymphoid lesions, we investigated 144 non-neoplastic lymphoid lesions by in situ hybridization (ISH) to detect the expression of EBV-encoded small RNAs (EBER)-1 and BCRF-1 and by immunostaining for latent membrane protein (LMP)-1 and ZEBRA. ISH for EBER-1 detected EBER-1-positive cells (EPC) in 31 of the 144 examined lesions (22%). EPC were detected in 4 of 49 cases of nonspecific lymphoid hyperplasia, in 16 of 20 abscess-forming granulomatous lymphadenitis (AFGL), 5 of 25 Kikuchi's disease, and in 3 of 3 infectious mononucleosis. LMP-1 was expressed in 6 of 124 non-neoplastic lymphoid lesions (4.8%). LMP-1-positive cells were observed in 6 of the 31 EBER-1-positive cases (19%). EPC were detected significantly more frequently in LMP-1- and ZEBRA-positive specimens than in the LMP-1- and ZEBRA-negative specimens. BCRF-1 was expressed in 4 of 11 cases examined: 2 of 3 AFGL, 1 of 2 Kikuchi's disease, and in the 1 case of atypical lymphoid hyperplasia. This study suggests that Epstein-Barr virus is prevalent and can be reactivated in the lymph nodes effaced by destructive inflammation, such as AFGL. Such inflammation may provide a local milieu that is conducive for EBV to enter the lytic cycle.

Many neurons in the adult rat cingulate cortex possess perineuronal sulfated proteoglycans detectable with cationic iron colloid and aldehyde fuchsin, or cell surface glycoproteins reactive to lectin Vicia villosa or soybean agglutinin. The perineuronal sulfated proteoglycans develop three to four weeks after birth. The cell surface glycoproteins develop at earlier stage or two to three weeks after birth. Dark or active neurons begin to appear three to four weeks after birth. These findings indicate that the brain matures after birth or during weaning period.

Cathepsin B, a thiol protease, is involved in cancer metastasis. To clarify the role of cathepsin B in tumor progression in human colorectal cancer, the relationship between its activity, immunohistochemical staining, and clinical tumor progression was investigated. Cathepsin B activity in adenocarcinomas was significantly elevated compared with that in the tumor-bearing tissue. Furthermore, the tumor/tumor-bearing tissue (T/Tb) ratio of the activity was significantly higher than that of colorectal adenoma. Immunohistochemical studies demonstrated intense staining in the cancerous tissue. With respect to the clinical stage of tumors, the activity tended to be higher in tumors that had invaded the serosa or subserosa than in those that invaded the proper muscle. The results suggest that cathepsin B participates in the progression of human colorectal cancer, and its increased expression is a sensitive marker of the differentiation between colorectal adenoma and adenocarcinoma.

Cathepsin B, a thiol protease, has been reported to be involved in cancer progression and metastasis. The suppressive effects of two kinds of protease inhibitors, leupeptin and dietary camostate (FOY-305), on tumorigenesis and progression in 1, 2-dimethylhydrazine (DMH)-induced rat colon neoplasm were examined in relation to tissue cathepsin B activity. Male Donryu rats were treated with leupeptin or FOY-305 during or after the administration of DMH. There were no significant differences in average tumor numbers among all DMH-treated groups. However, the percentage of small tumors was significantly higher in the group in which leupeptin was supplied during DMH administration. This trend was not recognized in the FOY-305-treated groups. The ratio of cathepsin B activity in the tumors to that in the tumor-bearing tissue (T/Tb) was significantly increased with increasing tumor size (P = 0.009). The cathepsin B activity levels in the tumor-bearing mucosa in the groups which received leupeptin or FOY-305 following DMH treatment were both significantly lower than that in the group which received neither protease inhibitor (P = 0.046 and P = 0.0067, respectively). The results obtained indicate that leupeptin may have suppressed tumor growth by lowering the tissue cathepsin B activity.

To clarify the relation between systemic and cutaneous vascular endothelial injury in progressive systemic sclerosis (PSS), we examined thrombomodulin (TM) expression in PSS skin lesions immuno-histopathologically and compared it with plasma soluble TM levels measured by specific enzyme-linked immunosorbent assay. The plasma soluble TM level in PSS patients was significantly higher than that of normal controls and was as high as the levels of SLE patients. In relation to disease activities, the plasma TM levels of sclerotic phase PSS patients were significantly higher than that of atrophic phase PSS patients. The plasma samples with anti-Scl-70 antibody showed a high TM level than samples with anti-centromere antibody or anti-RNP antibody. Barnett's types or systemic corticosteroid treatment did not affect the TM level. Histopathologically, the dermal endothelial TM expression significantly increased in the sclerotic skin and moderately increased in the non-sclerotic skin of PSS compared with that of normal control skin. In addition, immunoreactive TM expression in the epidermis also increased in PSS. Disease activity-dependent elevation of plasma TM levels and immuno-histopathological expression of TM suggested generalized endothelial and epidermal cell involvement in PSS, and compensation in part by overproduction of TM by endothelial cells.

A cross-sectional study was conducted to quantitatively evaluate the relationship between the instrumental activities of daily living (IADL) and various physical fitness tests in elderly women living at home. The study focused on the total population of those women aged 65 years and over living in Y Town, Hyogo Prefecture, Japan, who visited a nursing home for day services. A total of 128 subjects were divided into two groups: dependent in IADL group (n = 49) and independent in IADL group (n = 79). The magnitude of the relation was evaluated by the odds ratio (OR). The following tests showed a significant decrease in IADL: knee-raising test [age-adjusted OR = 4.23, 95% confidence interval (CI) 1.81-9.87], height (age-adjusted OR = 4.09, 95% CI 1.75-9.56), grip strength (age-adjusted OR = 3.68, 95% CI 1.57-8.60), sit-and-reach test (age-adjusted OR = 2.76, 95% CI 1.20-6.34), and standing on one leg with closed eyes (age-adjusted OR = 2.56, 95% CI 1.09-5.97). Multivariate analysis using Hayashi's quantification method I indicated that knee-raising was the test most highly correlated with decreased IADL. These results suggest that measurement of knee-raising ability, muscle strength of the lower extremities and flexibility of hip joint could be the most useful factors to assess the level of instrumental self-support ability.

Escape from cellular aging is the rate-limiting step of multistep carcinogenesis. While normal human cells invariably undergo cellular aging and almost never spontaneously immortalize, cells derived from rodents such as mice are relatively easily immortalized. In this experiment, we studied the immortalization patterns of cells obtained from brain tissues of an inbred strain (MSM/MSfB6C3F1) derived from wild mice. We established 12 cell strains derived from 12 mouse brains in order to investigate whether these cells show cellular aging in the same fashion as human cells or whether these cells are immortalized as easily as rodent cells reported previously. As a result, all cell strains were immortalized up to about 200 days in culture. One strain immortalized very early, in the first 50 days, four strains immortalized in the last 200 days, and the other seven strains became immortal between 150 and 200 days in culture. All immortalized cell strains showed varying amounts of chromosome abnormalities, numerically and structurally, but no specific changes related to immortalization were detected. Before immortalization, three types of cells, glial-like, polygonal flat-thin, and fibroblast-like cells, were observed in culture, but after immortalization most of the cultures became fibroblastic. From these results, we concluded that fibroblast-like cells derived from brains of these mice immortalized in like fashion to fibroblasts of other inbred mice.

Since platelets accumulate taurine, they provide a model for studying the taurine transport in anisosmotic disorders. Thus, in this work we studied the taurine concentration and uptake in the platelets of Brattleboro rats, homozygous (DI) and heterozygous (HZ) for hereditary hypothalamic diabetes insipidus, and Long Evans (LE) normal rats after free water intake and after dehydration induced by water deprivation for 24 h. The decreased ability of the DI rats to concentrate urine led to plasma hypernatremia and hyperosmolality despite excessive drinking. Water deprivation in the DI rats induced drastic dehydration with exacerbated hypernatremia and hyperosmolality. Plasma hypernatremia and hyperosmolality resulted in a significant elevation of the taurine concentration and uptake by platelets of the DI rats. Kinetic assays showed that plasma hypernatremia and hyperosmolality did not alter the affinity of taurine to platelet membrane carrier, as expressed by Km, but caused a profound increase in the maximal transport capacity, Vmax. After free water intake the Vmax of the DI rats was about two times higher than that in the HZ and LE rats and after water deprivation it was about three times higher. Water deprivation doubled the Vmax of the DI rats without changing the Km.

The use of ligand-binding methods to study neurotransmitter-receptor sites has made its impact on almost all aspects of biological pursuits including research on aging and neurodegenerative diseases. In the past, most of the research in biochemical gerontology has largely centered around changes in various neurotransmitters and enzymatic activities. The molecular basis of aging and neurodegeneration at the level of neurotransmitter-receptor interactions has been highly appreciated in the last two decades as a result of receptor binding studies. It is now possible to obtain information about the regional distribution of neurotransmitter receptors in the brain, the pharmacological and biochemical characteristics of these sites, and the functional interrelationships between different neuronal systems in normal and pathological conditions. The passage of time after maturity is accompanied by measurable physiologic decline in virtually all systems. It is the aim of this work to discuss the practical aspects of neurotransmitter and/or drug (ligand)-receptor binding studies, highlighting some examples of their applications to geriatric neuropharmacology research, with special consideration to learning impairment and memory loss in normal and in pathological aging processes.

We observed differences in the capillary architecture of the skeletal muscles that have different fiber metabolism. The soleus, the vastus intermedius and the tibialis anterior muscles of adult Wistar rats were prepared using two different techniques. Samples for adenosine triphosphatase (ATPase) staining were prepared following Dubovitz's method, and the distributions of fiber type, Types 1, 2A and 2B, were analyzed. Then, corrosion casts of capillary architecture of these muscles prepared following Murakami's method were observed with a scanning electron microscope (SEM) and compared with the fiber distribution. The fiber type composition of the soleus muscle showed Type 1 (slow-twitch) dominance and that of the vastus intermedius and the tibialis anterior muscle showed Type 2 (fast-twitch) dominance. The capillaries of the soleus muscle were tortuous, and this was thought to be advantageous for blood supply. In contrast, the capillaries of the vastus intermedius and tibialis anterior muscles had a relatively parallel pattern. Additionally, two different patterns of capillary architecture that appeared to correspond to certain metabolic characteristic of different muscle fiber types were preserved with corrosion casting. In conclusion, comparative studies on capillary architecture of the skeletal muscles are useful for analyses of its function.

In the pathophysiology of lumbosacral radiculopathy, inflammation of the nerve root is of critical importance. Additionally, free radicals have been shown to be associated with some inflammatory process. This study was designed to investigate whether free radicals participate in the pathophysiology of nerve root involvement. We measured superoxide dismutase (SOD) activity in cerebrospinal fluid (CSF) of 31 patients with unilateral lumbosacral radiculopathy caused by a herniated disc using electron spin resonance (ESR) spectrometry. Then SOD activity was compared with the type of nerve root compression as seen on preoperative myelography. SOD activity in the normal control group was 7U/ml, while that in the hernia group remarkably decreased. The concentration gradient of SOD activity was different between central herniation and centrolateral herniation. Our findings indicate that free radicals are generated after nerve root compression. Under severe deficiency of SOD activity in CSF, serum SOD penetrates into CSF after further compression. In addition, SOD in CSF may play an important role in protecting against nerve root involvement.

We conducted a clinicopathological analysis of chondrosarcomas in 17 patients treated in our institute. The 5- and 10-year overall survival rates of the patients were 72.3% and 61.9%, respectively. The significant prognostic factors were size and histologic grade of the tumor. Sex, age, location of the primary tumor, or the presence of a preceding exostosis did not affect the treatment results significantly. Chondrosarcomas of histologic grades I and II did not metastasize, while all grade III and dedifferentiated chondrosarcomas metastasized to the lung. The local recurrence rate depended on the surgical margin. Wide excision with an adequate surgical margin is important to achieve local control of the chondrosarcoma.

We report here the time-course of electron microscopic changes induced by gamma-interferon (IFN-gamma) in the human erythromyeloid leukemia cell line K562. In K562 cells treated with IFN-gamma for 6h, the nuclei were polygonal in shape and microvilli were far more abundant on cell membranes compared with control K562 cells, and invaginations were often seen in the cell membranes. There was a reduction in the number of cell-membrane microvilli and an increase in the number of lysosomal bodies in the cytoplasm of K562 cells treated with IFN-gamma for 12h. After treatment with IFN-gamma for 24h, the cell membrane microvilli disappeared, large numbers of cellular organelles were observed, such as mitochondria and lysosomes, and the cytoplasm became electron-dense. Cytoplasmic vesicles and vacuoles were also observed. These vesicles may correspond to an intermediate step in the ultimate cellular disintegration associated with apoptosis caused by IFN-gamma.

In order to evaluate a clinical use of omeprazole suspension, we examined the pharmacokinetics of omeprazole after oral administration in rats. Although the administration of omeprazole suspension buffered by NaHCO3 solution did not produce a significant increase in the area under the concentration-time curve (AUC) value compared with non-buffered group, the administration of NaHCO3 buffer immediately after dosing of omeprazole suspension buffered by NaHCO3 caused a significant increase in the AUC value. These results suggest that the NaHCO3 treatment following the administration of omeprazole buffered suspension effectively decreased the degradation of the compound by gastric acid. Therefore, the successive administration of NaHCO3 solution after the omeprazole dosing seems to be a simple and useful method for the administration to patients who cannot receive tablets.

Effects of HR-592, a new derivative of indole, on conditioned avoidance and intracranial self-stimulation behavior were investigated in rats using a shuttle box and a Skinner box, respectively. The oral administration of HR-592 at doses of 3-10 mg/kg caused a dose-dependent suppression of the conditioned avoidance response. Even the escape response was slightly suppressed in the group administered 10 mg/kg of HR-592. The self-stimulation behavior was suppressed dose-dependently from 1 to 8 h after the administration of 6-10 mg/kg of HR-592. These results indicate that the action of HR-592 on conditioned avoidance response and intracranial self-stimulation behavior is similar to the action of neuroleptics.