Low Amino Acid Score of Breakfast is Associated with the Incidence of Cognitive Impairment in Older Japanese Adults: A Community-Based Longitudinal Study.

BACKGROUND: The protein digestibility-corrected amino acid score (PDCAAS) represents the degree of utilizable dietary protein, namely the protein quality. The PDCAAS of a diet is required to be evaluated on a meal-by-meal basis, as food digestion and absorption occur with each meal intake. Although a positive association between protein intake and cognitive function has been reported, no study has investigated the association between PDCAAS of a diet and cognitive function. OBJECTIVES: To investigate the relationship between PDCAAS of a diet and cognitive impairment in older adults. DESIGN: Longitudinal epidemiological study. SETTING: Community-based setting. PARTICIPANTS: We analyzed 541 community-dwellers who participated in both baseline and follow-up survey. They were 60-83 years of age without cognitive impairment at baseline. MEASUREMENTS: Cognitive impairment was defined as a Mini-Mental State Examination (MMSE) score ≤27. Individual PDCAASs were calculated for each of three regular meals from the 3-day dietary records at baseline. Participants were classified into two groups according to the sex-specific tertiles (T1-T3) of the PDCAAS for each meal (i.e., T1 as the low score group and T2-T3 as the medium and high score group). The dependent variable was cognitive impairment observed after 4 years, and the explanatory variables were the PDCAAS groups for each meal (the medium and high group as the reference) and covariates (sex, age, body mass index, education, depressive symptoms, medical history, protein intake at each meal, and the MMSE score at baseline). Multivariable logistic regression analysis was performed to evaluate the low PDCAAS group for cognitive impairment after 4 years. RESULTS: A significant association was observed only between a low PDCAAS of breakfast and the incidence of cognitive impairment (the adjusted odds ratios [95% confidence intervals] of low PDCAAS for cognitive impairment for breakfast, lunch, and dinner were 1.58 [1.00-2.50], 0.85 [0.54-1.34], and 1.08 [0.71-1.65], respectively). CONCLUSION: A lower PDCAAS of breakfast, i.e., a diet with poor quality of protein, was associated with the incidence of cognitive impairment in older adults of the community.

The Japan-Multimodal Intervention Trial for Prevention of Dementia (J-MINT): The Study Protocol for an 18-Month, Multicenter, Randomized, Controlled Trial.

BACKGROUND/OBJECTIVES: The Japan-multimodal intervention trial for prevention of dementia (J-MINT) is intended to verify the effectiveness of multi-domain interventions and to clarify the mechanism of cognitive improvement and deterioration by carrying out assessment of dementia-related biomarkers, omics analysis and brain imaging analysis among older adults at high risk of dementia. Moreover, the J-MINT trial collaborates with partnering private enterprises in the implementation of relevant interventional measures. This manuscript describes the study protocol. DESIGN/SETTING: Eighteen-month, multi-centered, randomized controlled trial. PARTICIPANTS: We plan to recruit 500 older adults aged 65-85 years with mild cognitive impairment. Subjects will be centrally randomized into intervention and control groups at a 1:1 allocation ratio using the dynamic allocation method with all subjects stratified by age, sex, and cognition. INTERVENTION: The multi-domain intervention program includes: (1) management of vascular risk factors; (2) group-based physical exercise and self-monitoring of physical activity; (3) nutritional counseling; and (4) cognitive training. Health-related information will be provided to the control group every two months. MEASUREMENTS: The primary and secondary outcomes will be assessed at baseline, 6-, 12-, and 18-month follow-up. The primary outcome is the change from baseline to 18 months in a global composite score combining several neuropsychological domains. Secondary outcomes include: cognitive change in each neuropsychological test, incident dementia, changes in blood and dementia-related biomarkers, changes in geriatric assessment including activities of daily living, frailty status and neuroimaging, and number of medications taken. CONCLUSIONS: This trial that enlist the support of private enterprises will lead to the creation of new services for dementia prevention as well as to verify the effectiveness of multi-domain interventions for dementia prevention.

The Association between Dietary Amino Acid Intake and Cognitive Decline 8 Years Later in Japanese Community-Dwelling Older Adults.

OBJECTIVES: Previous studies have reported a relationship between low protein intake and cognitive decline and have suggested that this association may be related to specific amino acid intake. However, the effects of amino acid intake on the maintenance of cognitive function have yet to be clarified. We examined the longitudinal association between dietary amino acid intake and cognitive function in community-dwelling older adults. DESIGN: Longitudinal epidemiological study. SETTING: Community-based setting. PARTICIPANTS: This study comprised 427 study participants aged 60-82 years with no cognitive decline, defined as a Mini-Mental State Examination (MMSE) score of >27 at baseline, who also participated in a follow-up. The average and standard deviation of the follow-up period was 8.2 ± 0.3 years. MEASUREMENTS: Dietary intake was assessed using three-day dietary records at baseline. Participants were classified into quartiles (Q1-Q4) based on the intake of 19 amino acids for males and females. Next, we classified participants into Q1 and Q2-Q4 groups. Cognitive function was assessed using the MMSE both at baseline and at follow-up. Multivariable logistic regression models were used to estimate odds ratios (ORs) and 95% confidence intervals (CIs) for the associations between the Q1 group and cognitive decline (MMSE ≤27), using the Q2-Q4 group as a reference group. Covariates were age, sex, body mass index, years of education, severity of depressive symptoms, history of lifestyle diseases (hypertension, dyslipidemia, diabetes mellitus, stroke, and ischemic heart disease), energy intake (kcal/d), protein intake (g/d), and MMSE score at baseline. RESULTS: Cognitive decline was present in 133 (31.1%) participants. After adjustment for covariates, including total protein intake, the ORs (95% CIs) for cognitive decline were 2.40 (1.21-4.75) for lysine, 2.05 (1.02-4.09) for phenylalanine, 2.18 (1.09-4.34) for threonine, and 2.10 (1.06-4.15) for alanine. CONCLUSION: The results suggest that lysine, phenylalanine, threonine, and alanine intake is important for the maintenance of cognitive function in older people, independent of total protein intake.

Dietary Patterns and Muscle Mass, Muscle Strength, and Physical Performance in the Elderly: A 3-Year Cohort Study.

OBJECTIVES: Nutritional support effectively prevents and treats sarcopenia; however, the influence of overall dietary patterns on sarcopenia parameters is less investigated. This study aimed to determine the association between adherence to Mediterranean-style diet (MD), Dietary Approaches to Stop Hypertension (DASH), Japanese Food Guide Spinning Top (JFG-ST), and modified JFG-ST (mJFG-ST) and muscle mass, muscle strength, and physical performance in community-dwelling Japanese elderly. DESIGN AND SETTINGS: This prospective cohort study recruited individuals aged over 60 years from a community college in Nagoya, Japan. PARTICIPANTS AND MEASUREMENTS: A total of 666 participants were followed up annually from 2014 to 2017. Demographic data, anthropometric measurements, and sarcopenia parameters including walking speed (WS), hand grip strength in the dominant hand (HGS), and skeletal mass index (SMI) were recorded. Self-recall dietary intake was assessed using a validated food frequency questionnaire comprising 29 food groups. Adherence to MD, DASH, JFG-ST, and mJFG-ST was determined by tertiles. RESULTS: At baseline, the mean age of all participants (56.5% women) was 69.4±4.4 years. WS, HGS, and SMI were 1.4±0.2 (m/s), 28.9±8.1 (kg), and 6.7±1.0 (kg/m2), respectively. In longitudinal analysis, participants with higher JFG-ST adherence scores were more likely to have higher SMI (Q3 vs. Q1: mean difference, 0.048; p=0.04) after adjustment, and its benefits were more evident in men (Q2 vs. Q1: mean difference, 0.098; p=0.047; Q3 vs. Q1: mean difference, 0.091; p=0.017) than in women. WS and HGS were not associated with any type of dietary pattern. CONCLUSIONS: Adherence to JFG-ST was positively associated with SMI in Japanese community-dwelling elderly adults aged over 60 years, specifically in men. The country-specific dietary recommendations are required to be developed for sarcopenia prevention.

Combined Impact of Physical Frailty and Social Isolation on Rate of Falls in Older Adults.

OBJECTIVES: The aim of this study was to examine the impact of the combination of physical frailty and social isolation on falling in community-dwelling older adults. DESIGN: A cross-sectional study of data obtained at registration in a randomized control trial. SETTING: Community-based study of participants recruited from Toyota, Japan. PARTICIPANTS: 380 community-dwelling older adults (47.9% women, mean age = 72.3 ± 4.6 years). MEASUREMENTS: Participants were categorized as non-frail or pre-frail/frail based on the Fried frailty criteria (slowness, weakness, exhaustion, low activity, and weight loss). Social isolation was examined using the Lubben Social Network Scale (LSNS-6), and scores lower than 12 points indicated social isolation. Participants were divided into four groups depending on pre-frail/frail status and social isolation, and experiences of multiple falls over the past year were compared between the groups. RESULTS: Participants were classified into robust (n = 193), physical frailty (PF; n = 108), social isolation (SI; n = 43), and PF with SI (PF+SI; n = 36) groups. A total of 38 (10.0%) participants reported multiple falls. Logistic regression analysis showed that PF and SI groups were not independently associated with falling (PF: OR 1.64, 95% CI 0.65-4.16, SI: OR 2.25, 95% CI 0.77-6.58), while PF+SI group was significantly associated with falling compared with the robust group (OR 3.06, 95% CI 1.00-9.34, p = 0.049) after controlling for confounding factors. CONCLUSION: Our findings support the assertion that coexistence with physical frailty and social isolation were associated with falling in the older adults.

Cognitive Dysfunction in Urban-Community Dwelling Prefrail Older Subjects.

OBJECTIVES: A number of studies have reported that frailty is cross-sectionally associated with cognitive decline and is also a risk for future cognitive decline or dementia; however, there have been only a few studies that focus on the association between prefrailty and cognitive dysfunction. In the current study, we investigated the association between prefrailty and cognition. DESIGN: A cross-sectional study of the data obtained at registration in a randomized control trial. SETTING: Toyota, Japan. PARTICIPANTS: Community-dwelling older subjects (male 54.6%) who had cognitive complaints. MEASUREMENTS: A battery of neuropsychological and physical assessments were performed. Prefrailty was defined as exhibiting one or two of the five Fried criteria (weight loss, exhaustion, weakness, slow gait speed and low physical activity). We performed a multiple regression analysis to investigate the associations of cognitive performance with prefrailty, adjusting for the factors that were significantly different between the robust and prefrailty groups. To assess the cognitive attributes that were significantly associated with prefrailty, logistic analysis was performed to see if one specific criterion of the five frailty criteria was associated with cognitive performance. RESULTS: The study subjects included 183 prefrail and 264 robust individuals. The prefrail subjects with cognitive complaints were older, less educated, more depressive, and more likely to have diabetes mellitus than the robust subjects. The prefrail subjects had lower performance in a wide-range of cognitive domains, and after adjustments for age, education, depressive mood, and diabetes mellitus, prefrailty was associated with a decline in delayed memory and processing speed. Among the components of the Fried criteria, slow gait speed and loss of activity were significantly associated with slow processing speed as assessed by the digit symbol substitution test. CONCLUSION: The current results demonstrated that prefrailty was associated with worse memory and processing speed performance, but not with other cognitive domains.

Risk Factors for the Discontinuation of Home Medical Care among Low-functioning Older Patients.

OBJECTIVES: Older patients receiving home medical care often have declining functional status and multiple disease conditions. It is important to identify the risk factors for care transition events in this population in order to avoid preventable transitions. In the present study, therefore, we investigated the factors associated with discontinuation of home medical care as a potentially preventable care transition event in older patients. METHODS: Baseline data for participants in the Observational study of Nagoya Elderly with HOme MEdical (ONEHOME) study and data on the mortality, institutionalization, or hospitalisation of the study participants during a 2-year follow-up period were used. Discontinuation of home care was defined as admission to a hospital for any reason, institutionalization, or death. Univariate and multivariate Cox hazard models were used to assess the association of each of the factors with the discontinuation of home care during the observational period. The covariates included in the multivariate analysis were those significantly associated with the discontinuation of home care at the level of P<0.05 in the univariate analysis. RESULTS: The univariate Cox hazard model revealed that a low hemoglobin level (< 11g/dL), low serum albumin level (< 3g/dL), higher Charlson Comorbidity Index score, and low Mini Nutritional Assessment Short Form score (< 7) were significantly associated with the discontinuation of home care. A multivariate Cox hazard model including these four factors demonstrated that all four were independently associated with home-care discontinuation. CONCLUSIONS: The present results demonstrated that anemia, hypoalbuminemia, malnourishment, and the presence of serious comorbidities were associated with the discontinuation of home medical care among low-functioning older patients.

Factors associated with deterioration of mini nutritional assessment-short form status of nursing home residents during a 2-year period.

OBJECTIVE: A number of other studies have been conducted to verify the Mini Nutritional Assessment (MNA) or the MNA short form (MNA-SF) as a nutritional assessment/screening tool in various clinical settings or communities. However, there are few longitudinal studies using these tools to analyze which factors affect the incidence of deteriorating nutritional status. We tried to identify the factors associated with deterioration of MNA-SF status of nursing home residents during a 2-year period. METHODS: Participants were 392 people with a mean age of 84.3 in 12 nursing homes in Japan. The factors associated with deterioration in MNA-SF categories during the study period compared to stable/improved MNA-SF categories were identified. RESULTS: At baseline, 19.9% of the participants were malnourished and 60.2% were at risk of malnutrition, according to the MNA-SF classification. After 2 years, 66.3% participants maintained and 6.1% participants improved their nutritional status according to the MNA-SF classification, while 27.6% showed deterioration in MNA-SF status. Stepwise logistic-regression procedure indicated that basic ADL impairment and hospitalization during the follow-up period were associated with declining MNA-SF status. CONCLUSIONS: Poor basic ADL status and hospitalization during the follow-up period were associated with malnutrition and risk of malnutrition as assessed by MNA-SF of nursing homes residents during a 2-year period.

Preproghrelin Leu72Met variant contributes to overweight in middle-aged men of a Japanese large cohort.

OBJECTIVE: To investigate whether Leu72Met polymorphism of the preproghrelin gene is associated with overweight/obesity in middle-aged and older Japanese. DESIGN: Cross-sectional analysis. SUBJECTS: A total of 2238 community-dwelling middle-aged and older Japanese people (age: 40-79 years) who participated in the first wave of examinations in the National Institute for Longevity Sciences - Longitudinal Study of Aging from April 1998 to March 2000. MEASUREMENTS: The Leu72Met polymorphism of prepoghrelin gene, anthropometric variables including body weight, body mass index (BMI), waist circumference, waist-to-hip ratio and whole-fat mass and biochemical variables including serum lipid levels, fasting plasma glucose, insulin and homeostasis model assessment for insulin resistance. RESULTS: The frequencies of the Leu72Leu, Leu72Met and Met72Met alleles were 63.4, 32.7 and 4.0%, respectively. No differences in the genotype distributions of the Leu72Met polymorphism were found between genders or age groups, and no significant associations were observed between polymorphism and anthropometric variables in women and older men. However, middle-aged men who were 72Met allele carriers showed a higher body weight change from body weight at 18 years of age, as well as a higher waist circumference and a tendency to a higher waist-hip-ratio than noncarriers. Although there were no significant differences in the genotype distribution according to BMI in women and older men, a significantly higher frequency of the 72Met allele was found in the higher BMI group (BMI> or =25 kg/m(2)) of middle-aged men than in the normal-weight group. No significant associations were observed between polymorphism and serum lipid, glucose or insulin levels. CONCLUSIONS: These results suggest that the 72Met allele of the preproghrelin gene is a contributing factor for midlife weight change in men.

Powdery mildew (Podosphaera xanthii) resistance in melon is categorized into two types based on inhibition of the infection processes.

Physiological races of powdery mildew (Podosphaera xanthii) cause different symptoms in eight melon lines. Infection by races 1, 2, and 5 was examined in different melon lines. After a compatible reaction, conidia germination, haustorium initiation from the germ tube, germ tube branching, and sporulation occurred within 12, 24, 48, and 120 h, respectively, and the conidia matured within 240 h. In contrast, type i and ii inhibition were identified through incompatible reactions. The germ tube and haustorium were initiated from conidia, but no germ tube branching occurred in the lines with type i resistance within 48-240 h. In type ii resistance, germ tube branching was observed within 120 h, but no sporulation was observed within 240 h. The number of fluorescing epidermal cells was higher within 24 h in type i, and within 48-120 h in type ii resistance lines than in susceptible lines. Callose accumulation around the haustorium was detected in type ii resistance lines within 48-120 h. This suggests that the rapid hypersensitive response (HR) within 24 h has an important role in the type i response, while HR and callose accumulation in the type ii response occur slowly between 48 and 120 h. Of the resistant lines, PMR 45 and WMR 29 showed a type i incompatible response; the PI 414723 response was entirely type ii; and PMR 5, PI 124112, and MR-1 showed different responses depending on the race. Therefore, the two types of incompatible responses were intermixed in the same germplasm.

Powdery mildew (Sphaerotheca fuliginea) resistance in melon is selectable at the haploid level.

The major cause of powdery mildew in melons (Cucumis melo L.) is the fungus Sphaerotheca fuliginea. There are several cultivar- and season-specific races of this fungus. In order to control powdery mildew, it is important to introduce resistance to fungal infection into new cultivars during melon breeding. Haploid breeding is a powerful tool for the production of pure lines. In this study, it was investigated whether powdery mildew resistance could be manifested at the haploid level from two disease-resistant melon lines, PMR 45 and WMR 29. the effects of various races of S. fuliginea on diploid and haploid plants of PMR 45 and WMR 29 and of a disease-susceptible line, Fuyu 3 were measured. The responses of haploid and diploid plants to powdery mildew were identical. In addition, haploids that were generated from hybrids between Fuyu 3 and disease-resistant lines were examined. Seven out of 13 haploids from a Fuyu 3xPMR 45 cross and 10 out of 12 haploids from a Fuyu 3xWMR 29 cross were classified as resistant plants because they showed the same responses as their disease-resistant diploid parents to the various fungal races. These results indicate that resistance in PMR 45 and WMR 29 is selectable at the haploid level. All of the plant responses were observed by microscopy. A possible mechanism for generating powdery mildew resistance in two different melon lines is discussed.

Deficiency of the cysteine protease cathepsin S impairs microvessel growth.

During angiogenesis, microvascular endothelial cells (ECs) secrete proteinases that permit penetration of the vascular basement membrane as well as the interstitial extracellular matrix. This study tested the hypothesis that cathepsin S (Cat S) contributes to angiogenesis. Treatment of cultured ECs with inflammatory cytokines or angiogenic factors stimulated the expression of Cat S, whereas inhibition of Cat S activity reduced microtubule formation by impairing cell invasion. ECs from Cat S-deficient mice showed reduced collagenolytic activity and impaired invasion of collagens type I and IV. Cat S-deficient mice displayed defective microvessel development during wound repair. This abnormal angiogenesis occurred despite normal vascular endothelial growth factor and basic fibroblast growth factor levels, implying an essential role for extracellular matrix degradation by Cat S during microvessel formation. These results demonstrate a novel function of endothelium-derived Cat S in angiogenesis.

Glycation cross-links inhibit matrix metalloproteinase-2 activation in vascular smooth muscle cells cultured on collagen lattice.

AIMS/HYPOTHESIS: Extracellular matrix glycation has been proposed to contribute to the arterial stiffness observed in aging and diabetes. We examined whether matrix protein glycation regulates the proleolytic process through the manipulation of matrix metalloproteinases (MMPs) activation, using collagen fibrils model. METHODS: Vascular smooth muscle cells were cultured on control or glycated collagen fibrils. Matrix metalloproteinase-2 activation and the production of tissue inhibitors of metalloproteinase (TIMPs) were measured in the conditioned medium by using gelatin zymography and immunoblotting. Membrane type 1 matrix metalloproteinase (MT1-MMP) expression was also measured in cell lysates. RESULTS: When smooth muscle cells were cultured on collagen fibrils, pro-MMP-2 processing to active form was observed in the conditioned medium in coincidence with the increased MT1-MMP expression and the suppressed TIMP-2 production. Culturing smooth muscle cells on glycated collagen fibrils inhibited MMP-2 activation and attenuated MT1-MMP expression without the alteration of TIMP-2 production compared with control fibrils, indicating the possible mechanism of the suppression of MT1-MMP expression for the inhibition of MMP-2 activation on glycated collagen fibrils. Inclusion of aminoguanidine, an inhibitor of cross-linking formation, during collagen glycation restored the MMP-2 activation, suggesting the role of cross-links on the inhibition of MMP-2 activation. CONCLUSION/INTERPRETATION: These observations suggest that glycation-induced cross-linking formation in interstitial collagen contributes to arterial stiffness in aging and diabetes through the manipulation of matrix metalloproteinase activation along with the reduction of the susceptibility to proteolytic enzymes.

VEGF protects against oxidized LDL toxicity to endothelial cells by an intracellular glutathione-dependent mechanism through the KDR receptor.

Although the accumulation of vascular endothelial growth factor (VEGF) has been observed in human atherosclerotic lesions, the exact role of this growth factor in atherogenesis remains unknown. We hypothesized that VEGF in the vascular wall might have a preventive effect on endothelial cell damage during atherosclerosis. To test our hypothesis, we examined whether VEGF protects against the toxicity of oxidized low density lipoprotein (Ox-LDL) in cultured endothelial cells derived from bovine aortas (BAECs). Preincubation of BAECs with VEGF prevented Ox-LDL-induced toxicity in a preincubation time- and VEGF concentration-dependent manner. Addition of N(omega)-nitro-L-arginine methyl ester, a nitric oxide synthase inhibitor, did not reverse the protective effect of VEGF on Ox-LDL toxicity. Incubation of BAECs with VEGF increased intracellular glutathione (GSH) content in a time-dependent manner. Combined addition of VEGF and L-buthionine sulfoximine, a GSH synthesis inhibitor, reversed both GSH levels and the protective effect of VEGF on Ox-LDL-induced cytotoxicity. Placenta growth factor, which ligates to the VEGF Flt-1 receptor but not KDR/Flk-1, failed to prevent Ox-LDL toxicity and had no effect on intracellular GSH levels. An anti-KDR antibody completely blocked these beneficial activities of VEGF. These results suggest that VEGF prevents Ox-LDL-induced endothelial cell damage via an intracellular GSH-dependent mechanism through the KDR/Flk-1 receptor.

The effect of public long-term care insurance plan on care management and care planning in Japanese geriatric hospitals.

Japan, a society facing demographic aging at an incomparable speed, decided to introduce a new public long-term care insurance system to meet the expected need for elders to be cared for in the near future. The purpose of this study was to examine the change in knowledgeability on the side of managers of Japanese geriatric hospitals regarding (1) the concept of care-management as the methodology to supply care services to the elderly smoothly, and (2) comprehensive assessment for the elders as the technical skill in the process of materializing care management. Subjects were interviewed and questioned over the telephone according to a structural questionnaire. Managers (directors and office managers) of geriatric hospitals in 1996 and 1999. In 1996, the rate of knowledgeability for care management on the side of managers of geriatric hospitals was 70.0% (28/40) as a whole, increasing to 97.3% (71/73) in 1999. With regard to MDS-RAPs, the knowledgeability rate increased from 57.5% (23/40) in 1996 to 95.9% (70/73) in 1999. However, regarding care planning, almost all of the hospitals that performed care planning were those in the category required to submit care plans to the municipal government. Geriatric hospitals that were not required to submit such plans did not perform such planning either in 1996 or in 1999. It can be concluded that the decision to introduce a long-term care insurance system in Japan has led to a deeper understanding of the methodology of care management and a comprehensive assessment on the side of managers of geriatric hospitals.

[Estimation of radiation exposed area by the nuclear accident occurred at Tokai village using ESR measurements of household sugar].

The area of radiation exposure by the nuclear accident occurred at Tokai village in 1999 was estimated by the ESR measurement of 95 household sugar samples collected from the accident area. These samples were roughly classified into three types of sugar, fine white sugar, fine brown sugar and coarse brown sugar. The control fine white sugar showed no radical in the ESR spectrum, while those of fine brown sugar and coarse brown sugar showed the presence of a small amount of radicals. It was also shown that, among these three kinds of sugar, the radical concentration of fine white sugar sampled from wooden houses at the area similar to each other did not vary much with the samples, while those of fine brown sugar and coarse brown sugar varied to a considerable extent. Thus, the fine white sugar is considered to be more suitable for the estimation of the level of radiation exposure. The radical concentration of each fine white sugar sample was plotted against the distance from the site of the nuclear accident with a correction of the difference in the shielding effect between concrete houses and wooden houses. The samples obtained at more than 2 km north of the site of nuclear accident showed no ESR spectral signal to a detectable extent. On the other hand, the ESR spectra were observed from the samples obtained within 10 km south and 4 km west of the accident site. These results suggest that the radiation exposure by the contaminant blown by the northeast wind blowing on the day of the accident may occur at the south and west areas.

Rho-Rho kinase is involved in smooth muscle cell migration through myosin light chain phosphorylation-dependent and independent pathways.

Although Rho, a small GTPase, has been demonstrated to play an important role in the smooth muscle contraction and relaxation, little is known about the involvement of Rho protein in smooth muscle cell (SMC) migration. In this study the role of Rho-Rho kinase pathway was examined in SMC migration induced by platelet-derived growth factor (PDGF) and lysophosphatidic acid (LPA). C3 transferase, a specific inhibitor of Rho, blocked SMC migration induced by PDGF and LPA. Y-27632, a specific inhibitor of Rho kinase, a direct target molecule of Rho, inhibited PDGF and LPA-induced SMC migration in a concentration dependent manner. Although rapid increase in myosin light chain (MLC) phosphorylation in SMC treated with LPA was observed, no enhanced MLC phosphorylation was detected in response to PDGF. Y-27632 suppressed LPA-induced as well as basal level of MLC phosphorylation. ML-9, a specific inhibitor of myosin light chain kinase (MLCK), inhibited PDGF and LPA-induced SMC migration without the suppression of MLC phosphorylation at 5 min incubation, suggesting that MLCK may contribute to SMC migration via mechanism other than MLC phosphorylation. These results suggest that Rho-Rho kinase pathway is implicated in SMC migration and that different signaling pathways downstream of Rho-Rho kinase may be involved in LPA and PDGF-induced SMC migration. MLC phosphorylation via Rho-Rho kinase pathway appears to be implicated in LPA-dependent SMC migration. Whereas PDGF-mediated SMC migration is independent of increased MLC phosphorylation and other target molecules downstream of Rho-Rho kinase seem to be involved.