Levels of the oxidative stress marker γ-glutamyltranspeptidase at different stages of nonalcoholic fatty liver disease.

OBJECTIVES: This study investigated oxidative stress in the liver, by determining hepatic expression and serum levels of γ-glutamyltranspeptidase (GGT) and 8-hydroxy-2'-deoxyguanosine (8-OHdG) in different stages of nonalcoholic fatty liver disease (NAFLD), and assessed whether GGT can differentiate between the various stages of NAFLD. METHODS: Expression of GGT and 8-OHdG was examined in biopsy specimens by immunohistochemistry, and serum GGT and 8-OHdG levels were measured by enzyme-linked immuno sorbent assays in patients with simple fatty liver (n = 10), nonalcoholic steatohepatitis (NASH; n = 10) and, as a control, in alcoholic liver disease (ALD; n = 10). RESULTS: Hepatic tissue expression of GGT and 8-OHdG was seen in ALD, NASH and fatty liver patients. The percentage of hepatocytes positive for 8-OHdG expression and serum 8-OHdG levels was significantly higher in patients with NASH than simple fatty liver. Serum GGT levels were increased in all cases with ALD, NASH and fatty liver, and correlated significantly with serum levels of 8-OHdG in ALD and NASH, but not in simple fatty liver. CONCLUSIONS: Levels of GGT in fatty liver patients may compensate for mild oxidative stress by repressing 8-OHdG levels and preventing progression to NASH; however further oxidative stress leads to increased levels of 8-OHdG and the development of NASH.

PP121. Expression of PlGF, sFlt, MTF-1, HO-1 and HIF-1 alpha mRNAs in preeclampsia placenta and effect of preeclampsia sera on their expression of choriocarcinoma cells.

INTRODUCTION: Placenta growth factor (PlGF) is a growth factor originated from placenta. The sFlt-1 is soluble receptor for PlGF and suppresses PlGF function. It has been reported that in preeclampsia, serum level of PlGF decreased and sFlt-1 level increased and that preeclampsia placenta is in hypoxic condition. Metal-responsive transcription factor (MTF)-1, Hemoxigenase 1 (HO-1) and Hypoxia responsive factor -1 (HIF-1) may be induced in hypoxic condition. OBJECTIVES: In order to investigate pathophysiology in preeclampsia, we studied the expression of PlGF, sFlt-1, MTF-1, HO-1 and HIF-1 alpha mRNAs in placenta taken from preeclampsia and the effect of preeclampsia sera on their expression of choriocarcinoma cells and analysed the effect of placental hypoxia and serum factor on the expression of PlGF and sFlt-1 mRNA. METHODS: Placenta and serum samples were taken from preeclampsia and normal pregnancy with informed consent. The choriocarcinoma cells (JEG-3) were cultured in 24-well tissue culture plate. The cells were cultured with preeclampsia and normal pregnant sera. The RNAs were purified from these cells 24h after and placenta. The expressions of these mRNA were measured by using the real time PCR method (Applied Biosystems-7500). RESULTS: The expression of PlGF mRNA decreased and that of sFlt-1mRNA increased in preeclampsia placenta. The expression of MTF-1 and HO-1 mRNA decreased. The correlation was found between the expression of PlGF and MTF-1 mRNA, PlGF and HO-1 mRNA and sFlt-1 and HO-1mRNA. Moreover, expression of sFlt-1mRNA increased and HO-1mRNA decreased in JEG-3 cells after incubation of preeclampsia sera. CONCLUSION: The changes of PlGFmRNA in preeclampsia placenta may relate to the expression of MTF-1 and HO-1 mRNA. The changes of sFlt-1mRNA may relate to the expression of HO-1 mRNA and serum factor. Not only hypoxia but also serum factor may play a role of the levels of PlGF and sFlt-1 in preeclampsia placenta.

ACTN3 polymorphism affects thigh muscle area.

Muscle mass is an important factor influencing the activity of daily living in older adults. We aimed to investigate whether alpha-actinin-3 (ACTN3) gene R577X polymorphism affects muscle mass in older Japanese women. A total of 109 women (mean+/-SD, 64.1+/-6.0 years) were genotyped for the R/X variant of ACTN3. Mid-thigh muscle cross-sectional area (CSA) was assessed using MRI and compared using analysis of covariance models adjusted for body weight. In addition, physical activity and protein intake were measured as the living environmental factors affecting muscle mass. The ACTN3 R577X genotype distributions of the subjects were 19, 63 and 27 for the RR, RX, and XX genotypes, respectively. No differences in physical activity and protein intake were observed among the genotypes. The XX genotype showed lower thigh muscle CSA compared with RR&RX genotype (mean+/-SEM; XX: 69.1+/-1.8 cm(2), RR&RX: 73.6+/-1.1 cm(2); p<0.05). The results of the present study suggest that ACTN3 R577X polymorphism influences muscle mass in older Japanese women.

Effect of 12 weeks of moderate-intensity resistance training on arterial stiffness: a randomised controlled trial in women aged 32-59 years.

BACKGROUND: Resistance training has been increasingly incorporated into the overall exercise programme because of its effect on muscle strength, functional capacity and osteoporosis. High-intensity resistance training increases arterial stiffness. However, the effect of moderate-intensity resistance training on arterial stiffness is unknown. OBJECTIVE: To determine whether 12 weeks of moderate-intensity resistance training increases arterial stiffness in middle-aged women. METHODS: 35 middle-aged women (age range 32 to 59 years) volunteered to participate. The subjects were randomly assigned to one of three groups: resistance training (RT) group, aerobic exercise training (AET) group or control group. The RT and AET groups performed 12 weeks of moderate-intensity resistance training or aerobic exercise training (two days/week). RESULTS: In the RT group, one-repetition maximum strength significantly increased after the intervention. Interestingly, aortic (carotid-femoral) pulse wave velocity (PWV; an index of arterial stiffness), and peripheral (femoral-ankle) PWV did not change with moderate-intensity resistance training. In contrast, in the AET group, carotid-femoral PWV significantly decreased after the intervention. Resistance training and aerobic exercise training did not affect blood pressure. CONCLUSIONS: This study found that moderate-intensity resistance training did not increase arterial stiffness in middle-aged women, which may have great importance for health promotion with resistance training.

Ultrastructure of human placental tissue after 6 h of normoxic and hypoxic dual in vitro placental perfusion.

The dual in vitro perfusion model of human placental tissue allows the study of different aspects of placental function, such as metabolism, transport and secretion of proteohormones, cytokines and prostaglandins. The integrity of the perfused placental tissue is an important parameter to validate the perfusion system. Using light and electron microscopy, the morphology of villous tissue was examined before and after six hours of normoxic (n=10) vs. hypoxic (n=10) perfusion. An apical shift of the rough endoplasmic reticulum and occasional vacuoles were found in the syncytiotrophoblast of the terminal villi, the exchange area of the placenta. No unexpected pathological findings were seen before the perfusion experiments and only slight changes with moderate distension of the endoplasmic reticulum after 6 h of normoxic perfusion. After hypoxic perfusions, distinct ultrastructural alterations, such as oedematous villous stroma, swollen or completely destroyed cell organelles (e.g., mitochondria and endoplasmic reticulum), multiple vacuoles inside syncytio- and cytotrophoblasts as well as the microvilli were seen, which leads to an impairment of the placental barrier and other functions. The ultrastructural examination of placental tissue before and after dual in vitro perfusion broadens the knowledge of physiological and pathophysiological processes in the perfused placenta and may be a beneficial part of regular validation.

Effects of leg resistance training on arterial function in older men.

BACKGROUND: Little information is available on the effect of strength training on vascular function, particularly in older people. OBJECTIVE: To determine the effect of resistance training on arterial stiffness and endothelial function in older adults. METHOD: Eleven healthy men (mean (SEM) age 64 (1) years) performed 12 weeks of resistance training involving knee flexion and extension (three sets a day, two days a week). RESULTS: Resistance training increased maximal muscle power by 16% (p<0.0001). Arterial stiffness as assessed by aortic pulse wave velocity did not change with resistance training. Plasma concentration of nitric oxide (NO), measured as its stable end product (nitrite/nitrate), had increased (p<0.05) after resistance training (61.2 (10.4) v 39.6 (3.2) micromol/l). There was no change in plasma concentration of endothelin-1. CONCLUSION: The results suggest that short term resistance training may increase NO production without stiffening central arteries in healthy older men.

A 2-year follow-up study on muscle size and dynamic strength in teenage tennis players.

Growth trends in the cross-sectional area of the quadriceps femoris (CSA(QF)) and its dynamic strength in 12 teenage tennis players (six boys and six girls), aged from 10.7 to 13.2 years at the onset of the study, were investigated through a 2-year follow-up survey. CSA(QF) values at the three levels (proximal, mid, and distal to the knee joint) and dynamic torques during knee extensions at three pre-set velocities (1.05, 3.14, and 5.24 rad/s) were determined year by year, i.e., three times (T1, T2, and T3), using magnetic resonance imaging and an isokinetic dynamometer, respectively. In both genders, the CSA(QF) values at the three levels tended to increase across the measurement times, with greater gains in the boys than in the girls at the levels mid and distal to the knee joint. Among these changes, only the CSA(QF) at the level proximal to the knee joint significantly increased regardless of changes in both skeletal age and body height. The ratios of torque to the sum of CSA(QF) at the three levels (T/CSA) at 3.14 and 5.24 rad/s for the boys and at 5.24 rad/s for the girls were significantly higher in T2 and T3 than T1. Further, the relative increases in torque and T/CSA values at 3.14 and 5.24 rad/s were greater in the boys than the girls. The findings presented here indicate that young tennis players who are in the earlier stage of adolescence increase the CSA of the QF muscle beyond normally expected growth change at the level proximal to the knee joint and show a predominant development in torque generation capability during high-velocity knee extensions, with a greater gain in boys compared with girls.

Effect of spontaneous exercise on antioxidant capacity in rat muscles determined by electron spin resonance.

AIMS: The effect of physical activity on antioxidant capacity in muscle remains unknown. This study investigated the effect of spontaneous exercise on antioxidant capacity in rat muscles determined by electron spin resonance (ESR), which is a technique for the direct detection of free radicals. METHODS: Ten-week-old male Wistar rats were housed individually in cages with a running wheel. Rats were classified as high activity (HA), middle activity (MA) or low activity group (LA), based on an assessment of running distance covered over a 23-week period. After 23 weeks of housing, soleus (Sol), plantaris (Pl), gastrocnemius [deep/surface portions (GasD/GasS)] and heart (Hrt) muscles were isolated, and scavenging activity against superoxide anions (O(2)(*-)) and hydroxyl radicals (HO(*)) was determined by ESR using a spin-trap chemical. The citrate synthase (CS) activity was used as a marker of mitochondrial oxidative phosphorylation capacity. RESULTS: Among the parameters measured, only O(2)(*-) scavenging activity in GasD significantly correlated with the running distance. The highest scavenging activity was observed in Hrt of HA rats. The O(2)(*-) scavenging activity in Pl of MA rats was significantly higher than that of LA rats. The O(2)(*-) scavenging activity of Sol and GasS was not significantly different between the three groups. Furthermore, the HO(*) scavenging activity of any muscle specimens was similar among the three groups and did not correlate with running distance at all. CS activity did not significantly differ between the three groups. CONCLUSION: These data suggest that O(2)(*-) scavenging activity in specific types of muscle tissues would increase especially in spontaneously active animals. However, HO(*) scavenging activity would not.

Recruitment of the thigh muscles during sprint cycling by muscle functional magnetic resonance imaging.

The purpose of the present study was to investigate recruitment patterns of the thigh muscles during maximal sprint cycling by muscle functional magnetic resonance imaging (mfMRI). Twelve healthy men participated in this study and performed 2, 5, and 10 sets of 6-s supramaximal cycling with a load of 7.5 % of their body weight with 0.5 min of rest between the sets. Before and immediately after the exercise, T2-weighted MR images, i.e. mfMRI, of the right-thigh were taken to calculate T2 of eleven thigh muscles. Vastus lateralis, semitendinosus, and sartorius were the highest activated, i. e. had the greatest T2 change, among the quadriceps, hamstring, and adductors, respectively, compared with other muscles. Total power output during 2, 5, and 10 sets of sprint cycling was correlated with percent change in T2 in the quadriceps correlated (r (2) = 0.507 to 0.696, p < 0.01), the hamstring (r (2) = 0.162 to 0.335, p < 0.05 approximately 0.001), and the adductor muscles (r (2) = 0.162 to 0.473, p < 0.05 approximately 0.0001). With use of stepwise regression analysis, total power output was significantly correlated with % change in T2 of the vastus medialis (VM) (p < 0.0001) and vastus intermedius (VI) (p < 0.05) (r (2) = 0.698, p < 0.0001). We concluded that eleven thigh muscles were activated non-uniformly, and that the VM and VI play a key role during maximal sprint cycling.

Functional electrical stimulation (FES) for spinal cord injury.

Restoration of respiratory motion by stimulation of the phrenic nerve was investigated. Respiratory motion was restored successfully by introducing a breathing pacemaker to a patient with respiratory disturbance due to upper cervical spinal cord injury. Breathing pacemakers are considered to be more similar to physiological conditions compared to mechanical ventilators. Although the system is very expensive, its cost effectiveness may be excellent, provided that it can be used for long hours each day over an extended period. The system is effective in improving patient QOL because it dramatically increases patient mobility. From these findings, it is concluded that breathing pacemakers should be used more frequently in Japan, and that various forms of support are necessary to cope with economic and other concerns.

Muscle architectural characteristics in young and elderly men and women.

The purpose of this study was to compare the muscle architectural characteristics, i. e., muscle thickness, pennation angle, fascicle length, of four different groups comprising a total of 121 men and 190 women, divided according to sex and age (i. e., 20 - 39 yrs and 60 - 85 yrs). Muscle thickness and pennation angles of the vastus lateralis (VL), medial gastrocnemius (MG), long head of triceps brachii (TB) muscles were measured by B-mode ultrasonography, and fascicle length was estimated. Men had significantly greater relative muscle thickness (to limb length) than women, but not for MG. Relative muscle thickness of VL was significantly greater in younger subjects than in elderly (men; p < 0.001, women; p < 0.001), although there were no significant differences in relative muscle thickness of MG and TB between younger and elderly subjects. Men had significantly greater pennation angles than women, but not for MG in elderly subjects. The pennation angle of VL was significantly greater in younger subjects than in elderly (men; p < 0.001, women; p < 0.001), although there were no significant differences in pennation angles of MG and TB between younger and elderly subjects. Women had longer relative fascicle lengths (to limb length) of VL than men (p = 0.048 for younger, p = 0.028 for elderly). These results suggest that the decrease of thickness of the vastus lateralis muscle with aging is significant, and that there is the gender difference in the fascicle length of the vastus lateralis muscle.

Effects of 12 months of exercise training on salivary secretory IgA levels in elderly subjects.

BACKGROUND: The immune system declines in efficiency with advancing age, making the elderly less resistant to pathogenic microorganisms. Upper respiratory tract infection (URTI) is a common illness. Recent studies have shown that suppression of secretory immunoglobulin A (SIgA) is associated with increased incidence of URTI. OBJECTIVE: To assess the effect of exercise on salivary SIgA in elderly subjects. METHODS: Forty five elderly subjects (18 men, 27 women; mean (SD) age 64.9 (8.4) years) performed both 60 minute resistance and 60 minute moderate endurance training a week for 12 months. Saliva samples were obtained before training, and at four and 12 months during the training period. Salivary SIgA concentrations were measured by enzyme linked immunosorbent assay, and the SIgA secretion rate was calculated. RESULTS: SIgA concentrations before training, and at four and 12 months during training were 24.7 (14.4), 27.2 (14.2), and 33.8 (18.5) micro g/ml respectively. SIgA secretion rates were 29.5 (26.0), 33.8 (27.2) and 46.5 (35.1) micro g/min respectively. The results indicate that both the concentration and secretion rate of SIgA significantly (p<0.01) increased during 12 months of exercise in these elderly subjects. CONCLUSION: Regular moderate exercise seems to enhance mucosal immune function in elderly subjects.

Clinical and genetic studies of families with the tau N279K mutation (FTDP-17).

The tau N279K mutation was identified in four separately ascertained families in the United States, Japan, and France and in another recently discovered affected individual in Japan. The authors analyzed genealogical and clinical records and DNA samples. Average age at onset was 43 years; survival time was 7 years. All families exhibited similar clinical features, with parkinsonism, dementia, and supranuclear palsy uniformly seen. A founder effect indicated by a shared disease haplotype was seen only in two Japanese families. The N279K mutation can develop independently in different parts of the world.

Relationship between urinary symptoms and disease-related parameters in multiple sclerosis.

OBJECTIVE: To find the prevalence of voiding dysfunction in patients with multiple sclerosis and to examine the relationship between the voiding dysfunction and various parameters of the disease (disease severity, disease duration, lesion site, age and sex). METHODS: Using the International prostate symptom score, lower urinary tract symptoms were quantitatively evaluated in all patients with multiple sclerosis who had visited our neurological department during three months. The lesion site in the central nervous system was determined by a combination of neurological and magnetic resonance imaging findings. RESULTS: Of the 47 patients who had completed the questionnaire, 25 (53 %) were considered to have symptomatic voiding dysfunction. Even at early stage of the disability, 6 of 19 (32 %) patients were symptomatic. Eight (17 %) patients had irritative urinary symptoms alone, whereas 9 (19 %) patients had obstructive urinary symptoms alone. The irritative and obstructive symptoms were concomitant in 10 (21 %) patients. Compared with reports from Western countries, the ratio of obstructive symptoms to irritative symptoms was high in Japan. The degree of irritative symptoms was well correlated with the disease severity, whereas the correlation of obstructive symptoms with the disease severity was less significant. Irritative symptoms also showed a weak correlation with the disease duration, but obstructive symptoms did not. Among the lesion sites in the central nervous system, only the presence of the spinal cord lesion was related to the degree of urinary symptoms. Urinary symptoms were not significantly influenced by the age or the sex. Quality of life was disturbed by urinary symptoms, and this disturbance paralleled the disease severity. CONCLUSION: Urinary symptoms, especially irritative symptoms, reflect the condition of the disease. Thus, the quantified urinary symptoms may assist neurological diagnosis.

Relationship between mitochondrial DNA polymorphism and the individual differences in aerobic performance.

This study focused on the mitochondrial DNA (mtDNA) as the genetic factor most likely to bring about the individual difference in endurance capacity or its trainability. Platelets contain mtDNA but no nuclear DNA, whereas rho(0)-HeLa cells have nuclear DNA but no mtDNA. The oxidative capacity of mitochondria in the cultured cells, which were fused rho(0)-HeLa cell with platelets obtained from individual subjects (the so-called "cybrids"), reflects the individual mtDNA polymorphism in the gene-coding region. The purpose of this study was to investigate the relationship between the oxidative capacity of cybrids and the individual difference in endurance capacity, or its trainability. Forty-one sedentary young males took part in an 8-week endurance training program. They were determined by using their VO(2 max) as an index of endurance capacity on an ergocycle before and after the endurance training program. The relations between VO(2 max) before endurance training or the change of it by endurance training and the oxidative capacity of cybrids were investigated. There was no relation between them, and two groups were drawn from all subjects, based on one standard division of their initial VO(2 max): the higher pre-VO(2 max) group (n = 6) and the lower pre-VO(2 max) group (n = 5) (51.8 +/- 3.5 ml/min/kg vs. 33.3 +/- 3.8 ml/min/kg, p < 0.01). No significant difference was found between the O(2) consumption of the cybrids in the higher initial VO(2 max) group and that in the lower initial VO(2 max) group (16.3 +/- 4.9 vs. 15.9 +/- 2.0 nmol O(2)/min/10(7) cells, NS). Furthermore, neither the cytochrome c oxidase (COX) activity nor the complex I + III activity of cybrids showed a significant difference between the two groups. The oxidative capacity of cybrids between the high trainability group (n = 6) (Delta VO(2 max) 12.1 +/- 1.6 ml/min/kg) and the low trainability group (n = 9) (Delta VO(2 max) 2.3 +/- 0.5 ml/min/kg) was also similar. Thus the mtDNA polymorphism is very unlikely to relate to the individual difference in endurance capacity or its trainability in young sedentary healthy subjects.

Tumor necrosis factor gene polymorphisms in patients with sporadic Parkinson's disease.

We studied promoter region polymorphisms in the tumor necrosis factor (TNF) gene at position -1031, -863, and -857, in 172 Japanese patients with sporadic Parkinson's disease (PD). The frequency of the -1031C allele, a high producer of TNF, increased significantly in early onset PD patients compared with controls. In addition, PD patients with the -1031C allele showed a significantly earlier onset than those without -1031C allele, after stratification of the data by an interleukin-1beta gene polymorphism. Our findings suggest that TNF might have a toxic effect in PD.

Postural control responses sitting on unstable board during visual stimulation.

Concerning with the relation of vection induced by the optokinetic stimulation and the body movement, especially we attended to the neck joint movement, which counteracted to the shoulder movement. Then, we analyzed the mechanisms of the sitting postural control by using the seesaw board. By the optokinetic stimulation through the head mounted display (H.M.D.), the vection was leaded, and it affected to the sway of the body on the seesaw board. In this experiment, we found that the movement of upper part of body except for the head was the same direction to the seesaw board but the head moved out of phase to the seesaw board. This phenomenon will be suggested that the unstable condition of sway is balanced by the counter swing of head and the neck muscle tonus is controlled by acting of the vestibulo-collic reflex.

Riluzole stimulates nerve growth factor, brain-derived neurotrophic factor and glial cell line-derived neurotrophic factor synthesis in cultured mouse astrocytes.

Riluzole is an antiexcitotoxic agent used for the treatment of amyotrophic lateral sclerosis, and reported to have neuroprotective effects in animal models of Parkinson's disease, Huntington's disease and brain ischemia. We investigated the effects of riluzole on synthesis of nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF) and glial cell line-derived neurotrophic factor (GDNF) in cultured mouse astrocytes. The protein and mRNA levels were measured by enzyme-linked immunosorbent assay and semiquantitative reverse transcription-polymerase chain reaction, respectively. Treatment with riluzole at 100 microg/ml (426 microM) for 24 h increased the contents of NGF, BDNF, and GDNF in the culture medium 109-fold, 2.0-fold and 3.1-fold over the control, respectively. The drug-induced relative mRNA levels of NGF, BDNF, and GDNF were 7.3-fold at 2 h, 2.1-fold at 4 h, and 1.9-fold at 2 h, respectively. These results indicate that riluzole stimulates synthesis of NGF, BDNF and GDNF in cultured astrocytes. Riluzole might exert neuroprotective effects, at least in part, via stimulation of neurotrophic factors.

Endurance training under 2500-m hypoxia does not increase myoglobin content in human skeletal muscle.

The present study was carried out to determine whether myoglobin (Mb) concentration ([Mb]) in human skeletal muscle is influenced by 8 weeks of endurance training under normal conditions, and under hypoxic conditions equivalent to an altitude of 2500 m. Fourteen healthy but sedentary male adults who did not participate in any regular exercise program took part in this study. They were divided into two groups according to the training regime to which they were submitted: the N group, who exercised under normobaric conditions, and the H group, who exercised under hypobaric conditions. All subjects performed an incremental cycling exercise at sea level to evaluate their maximal O2 uptake (VO2max) before and after the 8-week endurance training course period. Muscle tissue samples were obtained by needle biopsy from the vastus lateralis muscle for histochemical and biochemical analysis. Training induced an increase in VO2max in both the N and H groups (P < 0.05), although there was no significant difference in these changes between groups. The 8-week training had no effect on [Mb] in either group. Muscle fiber composition was also unaffected by the training course. In contrast, citrate synthase activity in both groups increased by [mean (SD)] 28.2 (33.3)% (N: P < 0.01) and 32.0 (18.2)% (H: P < 0.05) after training, and the number of capillaries (capillary:fiber ratio) increased by 47.7 (33.8)% (N: P < 0.01) and 32.3 (20.6)% (H: P < 0.05). There were no significant differences in these parameters between the N and H groups. These results suggest that significant improvement of aerobic potential as a result of endurance training are not accompanied by increases in [Mb] in human skeletal muscle. In addition, a lower absolute workload may not be sufficient to stimulate Mb synthesis in humans, even where endurance training is carried out under hypoxia.