Developmental changes in expression of small GTPase RhoG mRNA in the rat brain

We have recently reported that RhoG, a member of Rho family small GTPases, is involved in neurite outgrowth in cultured neuronal cells. Here, we report the expression of RhoG mRNA in the developing rat brain by in situ hybridization analysis. At embryonic day 16, RhoG expression was observed throughout the ventricular zone, but was down-regulated in the region at birth. On the other hand, RhoG expression at postnatal day 20 was highly enriched in white matter tracts, including the corpus callosum, the anterior commissure, and the cerebellar white matter, and double-labeling experiments demonstrated that major RhoG-expressing cells in white matter tracts were oligodendrocytes. These results suggest distinct pre- and postnatal roles of RhoG in the development of the central nervous system.     

Involvement of apoptosis in the rat germ cell degeneration induced by nitrobenzene

Nitrobenezene (NB) produces germ cell degeneration, especially of spermatocytes in rats. To examine the possible involvement of apoptosis in this process, the extent and nature of nuclear DNA fragmentation after NB dosing were assessed using both terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) and DNA gel electrophoresis, in addition to conventional histological and electron microscopic procedures. Adult Sprague Dawley rats were treated with a single oral dose of NB (250 mg/kg) and euthanized subsequently at 6, 12, and 24 h and 2, 3, 5, and 7 days. The earliest morphological signs of germ cell degeneration in testes were found in pachytene spermatocytes 24 h after dosing. Electron micrographs of degenerating spermatocytes showed marked nuclear chromatin condensation at the nuclear periphery and crowding of cytoplasmic constituents, which are characteristic of apoptosis. Coincident with the appearance of such morphological changes, degenerating spermatocytes contained fragmented DNA as revealed by TUNEL. The presence of DNA laddering, a hallmark of apoptosis on gel electrophoresis, was first apparent and most prominent at 24 h, gradually becoming less detectable. No such changes were observed up to 12 h after dosing or in control animals. These results demonstrated unequivocal involvement of apoptosis in the induction of germ cell degeneration caused by NB. Received: 29 September 1997 / Accepted: 6 January 1998     

High single-dose alternate-day corticosteroid regimens in treatment of polymyositis

Summary High single-dose alternate-day prednisolone therapy (ADT) was compared with daily-dose prednisolone therapy (DDT) for treatment of polymyositis. Thirty patients with polymositis were treated with ADT for an average of 33.9 months. The combined number of improvements was 21 out of 30, a response rate of 70%. Side-effects were very rare and mild. On the other hand, 9 of 17 patients treated with DDT for an average 18.5 months improved, a response rate of 53%. The incidence of side-effects was strikingly higher than with ADT. ADT is therefore strongly advocated for treatment of polymyositis to avoid infectious complications and lessen cushingoid side-effects.     

Cellular kinetics and K-ras codon 12 mutations according to histomorphometric type of colorectal polyps with epithelial serrated proliferation

Recent studies have examined cellular kinetics and genetic abnormalities in colorectal polyps with epithelial serrated proliferation (CP-ESP), including hyperplastic polyps, serrated adenomas, and tubulovillous adenomas. However, difficulty in histologically classifying these lesions and the lack of clear-cut diagnostic criteria have led to inconsistent findings. Some 60 cases of CP-ESP and 6 cases of CP-ESP with malignant transformation were studied. When nuclear size progressively decreased from the bottom and middle layers to the surface layer of the crypts, maturation gradient was considered positive. CP-ESP and CP-ESP adjacent to carcinoma were morphologically classified as being positive or negative for maturation gradient and inferior and lateral glandular branching. Cellular kinetics were evaluated using Ki-67 immunostaining, and polymerase chain reaction (PCR)-dependent preferential homoduplex formation assay was performed to detect the presence or absence of K-ras codon 12-point mutations. CP-ESPs were morphologically classified into five types. In CP-ESP of types 1 to 4, the proliferation zone was confined to the bottom layer of the crypts or extended from the bottom to middle layers. In contrast, the proliferation zone extended throughout the crypts in most type 5 lesions. K-ras codon 12 mutations were detected in only types 3 and 5 CP-ESP. The five histomorphologic types of CP-ESP have distinct patterns of cellular kinetics. Histomorphologically, type 3 CP-ESP is considered an atypical hyperplastic polyp, occasionally associated with an elongated proliferation zone or K-ras mutations. Preliminary evidence indicates some relation between K-ras mutations and structural atypia associated with lateral branching of the crypts.     

Hepatic CCR7lowCD62LlowCD45RClow allograft dendritic cells migrate to the splenic red pulp in immunologically unresponsive rats

Donor dendritic cells (DC) migrate into the recipient spleen after hepatic transplantation. Immunological unresponsiveness to rat hepatic allografts can be induced by prior donor-specific blood transfusion (DST). We investigated homing receptor phenotype and splenic distribution of donor DC after allografting and DST. Immunostaining revealed OX62+ cells in the splenic red pulp of animals receiving pre-transplant DST but only in the white pulp of untreated animals. Most OX62 cells were positive for OX76. There were two subsets of DC in the spleen, CD45RChighOX62+ and CD45RClowOX62+ cells. RT-PCR revealed that CD45RClowOX62+ cells expressed interleukin (IL)-10, while CD45RChighOX62+ cells expressed IL-2 and low levels of IL-10 mRNA. CD45RChighOX62+ cells strongly expressed CCR5 and CCR7, compared with weak expression in CD45RClowOX62+ cells. The Epstein-Barr virus-induced molecule 1 (EBI-1) ligand chemokine (ELC/MIP3beta) was expressed mainly within the splenic white pulp. Mucosal vascular addressin-cell adhesion molecule-1 (MAdCAM-1) was expressed in the marginal zone and white pulp, but expression of splenic MAdCAM-1 was down-regulated in DST-treated animals. L-selectin (CD62L), the ligand for MAdCAM-1, was strongly expressed on CD45RChighOX62+ cells but not on CD45RClowOX62+ cells. In conclusion, differential splenic migration of CCR5lowCCR7lowCD62Llow CD45RClow DC expressing Th2-type cytokines is associated with immunological unresponsiveness to rat hepatic allografts.     

Weight gain in childhood and bone mass in female college students

The attainment of maximal peak bone mass early on in life is one of the most important strategies for the prevention of osteoporosis in women. The aim of this study was to clarify the correlation between gains in body size in all growth phases in childhood and adult bone mass in women. The subjects were 86 female first-year university students, aged 18–21 years. We measured the subjects bone mineral content (BMC) and bone mineral density (BMD) at the lumbar spine and the left hip, including the femoral neck, with dual energy X-ray absorptiometry. Each subject was measured for current height and weight. Height and weight at birth, and at 1.5, and 3 years were obtained from each maternity record book, and those between 6 and 18 years were obtained from their school health records. Other information, including physical activity and calcium intake, was obtained through an interview. Bivariate analysis showed that weight gains during the periods from birth to 1.5 years and from 9 to 12 years significantly correlated with both BMC and BMD values at any site. The stepwise method of multiple regression analysis showed that a weight gain during the period from birth to 1.5 years was significantly associated with BMC at the lumbar spine (P = 0.0001) and at the femoral neck (P = 0.0290) and with BMD at the lumbar spine (P = 0.0387). Birth weight was significantly associated with BMC at the lumbar spine (P = 0.0474) and the total hip (P = 0.0352), and weight gain during the period from 9 to 12 years was significantly associated with BMC at the femoral neck (P = 0.0376). In conclusion, birth weight and weight gain in infancy are important determinants of bone mass in young women. Additionally, a girls prepubertal growth spurt is likely to be a key phase for the acquisition of bone mass in relation to body weight. Our findings suggest that osteoporosis prevention programs may need to start very early in the life cycle.     

Effects of Japan Sea Proper Water on the growth ofLegionella pneumophila, Escherichia coli, andStaphylococcus aureus

Objective To assess whetherLegionella pneumophila serogroup 1 and serogroup 6,Escherichia coli, andStaphylococcus aureus can survive in Japan Sea Proper Water (JSPW). Methods The inhibitory effects of JSPW, surface seawater (SSW), phosphate buffer solution with 3.5% NaCl of pH 7.0 (3.5% NaCIPBS), and the 10²- and 10⁴-fold dilute solutions with purified water or phosphate buffer solution of pH 7.0, and purified water were investigated. Survival cells were counted immediately after the water and the bacteria were mixed, and at 1,3,5, and 7 days after incubation at 37°C. If the number of surviving cells was decreased more than 2 log units compared with the starting value, we judged the medium to have had an inhibitory effect on the growth of the bacteria. Results The survival cells of the bacteria in JSPW had decreased more than 2 log units compared with the starting value at 1 day after incubation. After 1 day of incubation, the cells ofLegionella pneumophila serogroup 6 andStaphylococcus aureus were found to have decreased more than 2 log units in purified water (PW) used as a control. Furthermore,Legionella pneumophila serogroup 1 in the 10²-fold dilute solution of JSPW was only 1.04 log units lower than the starting value at 7 days after incubation. In the 10²- and 10⁴-fold dilute solutions of JSPW,Escherichia coli survived for 7 days after incubation. These results were almost similar to the results in SSW and 3.5% NaCIPBS. Conclusions The present findings demonstrate thatLegionella pneumophila serogroup 1 andEscherichia coli cannot survive in undiluted JSPW for over a day at 37°C, suggesting the inhibitory effects may be due to the sodium chloride contained in JSPW.     

Health characteristics of elderly Japanese requiring care at home

The number of elderly subjects requiring care is rapidly increasing, however, their health status has not been well studied. The purposes of this study were to describe the health characteristics of the elderly at home who were using long-term care insurance, and to clarify factors that influence dependence for activities of daily living (ADL). The subjects were 194 elderly people living at home, who were approved for care. The items surveyed were the demographic characteristics, care level (or level of dependence for ADL), Barthel index, grip strength, thigh muscle volume, cognitive impairment using the mini-mental state examination (MMSE), depressed mood using the geriatric depression scale (GDS-15), and serum albumin and hemoglobin levels. The grip strength (p=0.0001), thigh muscle volume (p=0.0030), MMSE score (p=< 0.0001) and serum albumin level (p= < 0.0001) decreased, while the GDS-15 score (p=0.0142) increased with deteriorating care levels. The proportion of subjects not requiring assistance for the items "bathing" and "stair-climbing" in the Barthel index was markedly low in the subjects belonging to the relatively low (mild) care levels. The logistic regression analysis showed that factors associated with dependence for "bathing" and "stair-climbing" were the use of day-services, male sex, decline of grip strength, and a high GDS-15 score. In conclusion, it is important to strengthen the muscles of the upper half of the body, and to correct depressed moods for maintaining levels of ADL in the elderly requiring care at home.     

Factors affecting short-term mortality in very low birth weight infants in Japan

No epidemiological surveys have examined risk factors related to the death of very low birth weight infants (VLBWIs) in Japan. The objectives of this study were to examine the death rate and fatalities related to complications among VLBWIs, and to analyze factors possibly determining the death of VLBWIs. The subjects of this study were 811 VLBWIs admitted to the Neonatal Care Center of Niigata City General Hospital between April 1987 and March 2003. We obtained information on gender, birth weight, gestational age, Apgar scores, single/multiple pregnancy, postnatal transfer, mode of delivery, complications and outcome (alive or deceased) at the time of discharge from medical records. Of the 811 infants, 98 died prior to discharge (12.1%). Logistic regression analysis showed that independent risk factors for death of VLBWIs were male gender (relative risk [RR]: 2.0), low birth weight (RR: 0.56), necrotizing enterocolitis (RR: 58.0), pulmonary hypoplasia (RR: 37.8), chromosomal abnormalities (RR: 36.3), congenital heart diseases (RR: 9.8), persistent fetal circulation (RR: 9.6), neonatal asphyxia (RR: 6.3) and sepsis (RR: 4.4). The risk for death rises 1.8-fold if birth weight decreases by 100 g. A very high risk of perinatal death is associated with necrotizing enterocolitis, pulmonary hypoplasia or chromosomal abnormalities. The risk of death due to congenital heart diseases or neonatal asphyxia is relatively lower, but the incidences of these two disorders are high (8% and 6%, respectively). From the viewpoint of prophylactic treatment aimed at reducing the death rate of VLBWIs, measures to increase birth weight are of primary importance. Furthermore, early treatment and improved perinatal management of congenital heart diseases and neonatal asphyxia are anticipated to reduce the overall death rate of VLBWIs.