Virus activation and immune function during intense training in rugby football players.

Epidemiological studies suggest that highly trained athletes are more susceptible to upper respiratory tract infections (URTI) compared with the general population. Upper respiratory symptoms (URS) often appear as either primary invasion of pathogenic organisms and/or reactivation of latent viruses such as Epstein-Barr virus (EBV). The purpose of this study was to examine the relationship between EBV reactivation and the appearance of URS during intensive training in collegiate rugby football players. We evaluated EBV-DNA expression in saliva and examined the relationship between onset of URS and daily changes in EBV-DNA as well as secretory immunoglobulin A (SIgA) levels among 32 male collegiate rugby football players during a 1-month training camp. The EBV-DNA expression tended to be higher in subjects who exhibited sore throat (p=0.07) and cough (p=0.18) than that of those who had no symptoms, although their differences were not significant. The SIgA level was significantly lower 1 day before the EBV-DNA expression (p<0.05). The number of URS increased along with the EBV-DNA expression and decrease of SIgA levels. These results suggest that the appearance of URS is associated with reactivation of EBV and reduction of SIgA during training.

A major quantitative trait locus controlling cadmium translocation in rice (Oryza sativa).

The trait of low cadmium (Cd) accumulation in brown rice (Oryza sativa) is important for food safety. An effective way to reduce Cd accumulation in the grain is to control Cd transfer from the roots to the shoots. Here, we investigated genotypic variation in the shoot Cd concentration among 146 accessions from a rice core collection and performed a quantitative trait locus (QTL) analysis to determine the loci controlling shoot Cd accumulation. Furthermore, we physiologically characterized the two accessions used for QTL analysis. Large genotypic variation (13-fold) in the shoot Cd concentration was found. A major QTL was detected on chromosome 11 using a F2 population derived from Badari Dhan (a high-Cd accession) and Shwe War (a low-Cd accession). This QTL explained 16.1% of the phenotypic variation in Cd accumulation. Furthermore, this QTL was confirmed by analysis of advanced progeny. Physiological studies showed that Badari Dhan and Shwe War did not differ in uptake of Cd by the roots, but differed greatly in the translocation of Cd from the roots to the shoots. Taken together, our findings suggest that the major QTL detected is responsible for the translocation of Cd from the roots to the shoots.

Novel QTLs for photoperiodic flowering revealed by using reciprocal backcross inbred lines from crosses between japonica rice cultivars.

The rice japonica cultivars Nipponbare and Koshihikari differ in heading date and response of heading to photoperiod (photoperiod sensitivity). Using simple sequence repeat (SSR) and single nucleotide polymorphism (SNP) markers, we conducted quantitative trait locus (QTL) analyses for heading date in a set of reciprocal backcross inbred lines (BILs) from crosses between Nipponbare and Koshihikari. Under natural-day conditions, transgressive segregation in days to heading (DTH) toward both early and late heading was observed in both BIL populations. QTL analyses revealed that two QTLs--on chromosomes 3 and 6--were involved in the difference in heading date between the parental cultivars. The Nipponbare allele at the QTLs on chromosomes 3 and 6 showed, respectively, increasing and decreasing effects on DTH in both BIL populations. The transgressive segregation observed in the BILs could be accounted for mainly by the complementary action of a set of alleles with opposing effects. Both QTLs were finely mapped as single Mendelian factors in secondary mapping populations (BC2F2 plants/BC2F3 lines). The QTL on chromosome 3 was mapped in the 1,140-kb interval between 94O03-4 (SSR) and OJ21G19-4 (SNP) and was designated Hd16. The QTL on chromosome 6 was mapped in the 328-kb interval between P548D347 (SSR) and 0007O20 (SSR) and was designated Hd17. Both Hd16 and Hd17 were involved in photoperiod sensitivity, as revealed by observation of the DTH of nearly isogenic lines of Nipponbare under short- and long-day conditions, suggesting that allelic differences in both Hd16 and Hd17 account for most of the difference in photoperiod sensitivity between the parental cultivars.

A rat model of saliva secretory immunoglobulin: a suppression caused by intense exercise.

We aimed to develop a valid model of immunosuppression induced by intense exercise in rats. Rats were divided into three groups. In the rest (Rest) group, saliva was collected from resting rats on 4 consecutive days. In the exercise (Ex) group, rats ran on a treadmill until exhaustion (exercise time: 60.0 +/- 3.7 min), and their saliva was collected before and after exercise; the salivary glands were removed after exercise. In the control (Con) group, saliva collection and gland removal were also performed, but the rats did not exercise. Secretory immunoglobulin A (SIgA) concentrations in saliva and polymeric immunoglobulin receptor (pIgR) mRNA expression in the glands were measured. There was no significant change in SIgA concentration in the Rest group over 4 days. In the Ex group, SIgA concentration decreased significantly after exercise compared with before, whereas there was no significant change in the Con group. The expression of pIgR mRNA was significantly lower in the Ex group post-exercise than in the Con group. Our procedure for saliva collection appeared suitable, and the exercise-induced SIgA suppression was probably caused by a decline in pIgR mRNA expression. We propose to use this reproducible and reliable rat model of exercise-induced SIgA suppression in future studies.

Effects of music during exercise on RPE, heart rate and the autonomic nervous system.

AIM: The purpose of this study was to investigate the relationship between the influence of music on RPE during sub-maximal exercise and on the autonomic nervous system before and after sub-maximal exercise. METHODS: Heart rate (HR), HR variability (HRV) and rates of physical fatigue (RPE) during exercise at 60% and at 40% VO2max with and without music were measured. The exercise protocol consisted of a 30-min seated rest (control) period followed by a 30-min submaximal cycling exercise and a 35-min recovery period. Autonomic-nervous activity was measured before and after exercise. During exercise, RPE was recorded every 3 min and HR was recorded for every minute. RESULTS: Although RPE did not differ during exercise at 60% VO2max, this value was lower during exercise at 40% VO2max in the presence, than in the absence of a favorite piece music (P < 0.05). HR, HFA and LFA/HFA of HRV significantly differed with exercise intensity in the absence (P < 0.05), but not in the presence of music. CONCLUSIONS: These findings suggested that music evokes a ''distraction effect'' during low intensity exercise, but might not influence the autonomic nervous system. Therefore, when jogging or walking at comparatively low exercise intensity, listening to a favorite piece of music might decrease the influence of stress caused by fatigue, thus increasing the ''comfort'' level of performing the exercise.

Changes of pituitary, adrenal and gonadal hormones during competition among female soccer players.

AIM: This study was intended to investigate various hormones during competition among female athletes. METHODS: Subjects were elite female soccer players (n=9). Blood samples and profiles of mood states (POMS) were obtained before (Pre), during 3 days of competition (Competition), and after competition (Post-3 days). Serum concentrations were determined for dehydroepiandrosterone sulfate (DHEAS), cortisol (C), prolactin (PRL), testosterone (T), adrenocorticotropic hormone (ACTH), luteinising hormone (LH), and follicle stimulating hormone (FSH). RESULTS: Levels of C and PRL increased significantly during competition (2nd: P<0.05; 3rd: P<0.01) compared with Pre. Levels of LH increased significantly during competition (3rd day) compared with Pre (P<0.05). Levels of DHEAS changed significantly during competition, whereas levels of T, ACTH, and FSH were unchanged during competition. The fatigue score of POMS increased significantly during competition (2nd day) compared with Pre (P<0.05). CONCLUSIONS: We demonstrated that competitive stress affected hormonal status in female athletes. These findings suggest that hormonal changes reflect physical and mental stress during competition.

Daily changes of salivary secretory immunoglobulin A and appearance of upper respiratory symptoms during physical training.

AIM: It is well known that highly trained athletes suffer from a high incidence of upper respiratory tract infections (URTI). Secretory immunoglobulin A (SIgA) is a major effector of mucosal surface protection against microorganisms causing URTI. Although several studies have investigated the relationship between falls in SIgA levels and appearance of URTI symptoms, the relationship is not yet clear. METHODS: We prospectively investigated the relationship between daily changes in SIgA and appearance of URTI symptoms in collegiate soccer players during a training period of 2 months. RESULTS: Five of 12 subjects exhibited URTI symptoms during the study period. The SIgA level did not significantly decrease before appearance of URTI symptoms. However, the saliva flow rate and SIgA secretion rate tended to decrease 3 days before the appearance of URTI symptoms compared to that in the non-infection period (31.3+/-19, -42.2+/-20.6%, respectively). CONCLUSIONS: We could not demonstrate a significant relationship between decreased SIgA levels and appearance of URTI symptoms during the training period. However, our findings suggest that monitoring of SIgA secretion rate may be useful for assessment of risk status of athletes for URTI.

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.

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.

Antiprothrombin autoantibodies in severe preeclampsia and abortion.

PURPOSE: We examined the levels of autoantibodies against prethrombin-1 and fragment-1 in pregnant women to determine the type of autoantibodies that were associated with severe preeclampsia or spontaneous abortion. SUBJECTS AND METHODS: We measured autoantibodies bound to prothrombin, prethrombin-1, and fragment-1 by using an enzyme-linked immunosorbent assay (ELISA) in 12 healthy nonpregnant women, 36 women with normal pregnancies, 28 pregnant women with severe preeclampsia, and 19 pregnant women who subsequently had spontaneous abortion. RESULTS: Plasma samples in 10 (36%) of the 28 women with severe preeclampsia and 11 (58%) of the 19 women with spontaneous abortion were positive for antiprothrombin antibodies as compared with 3 (9%) of the 36 women with normal pregancies. All 11 of the positive samples from women who had spontaneous abortions were positive for antiprethrombin-1 antibody, but only 1 was positive for antifragment-1 antibody. The mean (+/- SD) titer of antiprethrombin-1 antibodies in patients with spontaneous abortion (36 +/- 9 U) was higher than that in women with normal pregnancies (10 +/- 4 U; P < 0.01). Antiprethrombin-1 antibody was detected in only 2 women with severe preeclampsia, whereas all 10 women with antiprethrombin antibodies were positive for antifragment-1 antibody. The antifragment-1 antibody titer in patients with severe preeclampsia (49 +/- 15 U) was higher than in women with normal pregnancies (13 +/- 6 U, P < 0.01). CONCLUSIONS: There is a strong and specific association between various types of antiprothrombin antibodies with severe preeclampsia and spontaneous abortion.

Changes in gamma-aminobutyric acid content during beni-koji making.

The changes in the gamma-aminobutyric acid (GABA) content during the making of beni-koji prepared with Monascus pilosus IFO 4520 vs. the difference in the rate of tomo koji (10%, 30%, and 50%) were examined. The increased proportion of tomo koji would increase the GABA production and the productions of GABA peaked on the fifth day and thereafter declined. The glutamate decarboxylase activity during beni-koji making with 50% tomo koji steadily increased after the start of the koji making, reaching its peak on the fifth day. The succinic acid content increased after the sixth day. The mycelial growth was in the stationary phase after the sixth day. Therefore, the GABA content increases with an increase in the proportion of tomo koji. It is presumed that the maximum amount of GABA reached on the fifth day was the cause of the increasing amount of conversion of GABA into succinic acid, in addition to the decline in the GAD activity after the fifth day of koji making.

Effect of brief maximal exercise on circulating levels of interleukin-12.

Interleukin-12 (IL-12) is a cytokine that was originally identified as natural killer cell stimulatory factor. It induces the activity of T-helper 1 (Th1) cells and exhibits strong anti-tumor activity. In this study, we studied the effects of brief anaerobic maximal exercise on circulating levels of IL-12. Six healthy males [mean (SD) 25.2 (2.6) years] performed a modified Wingate test exercise (resistance 0.075 kg/kg of body mass). The exercise consisted of five bouts of maximal cycling for 10 s, with rest intervals of 50 s between them. Blood samples were taken before, immediately after, 30 min after, 60 min after and 120 min after the exercise. Plasma concentrations of IL-12 were measured using an enzyme-linked immunosorbent assay. Data were corrected for hemoglobin and hematocrit measurements. Plasma concentrations of IL-12 averaged [mean (SD)] 234.2 (40.9) pg/ml before, 305.2 (62.1) pg/ml immediately after, 202.8 (24.2) pg/ml 30 min after, 239.7 (35.1) pg/ml 60 min after, and 199.6 (49.2) pg/ml 120 min after the exercise. We showed that plasma concentrations of IL-12 increased significantly immediately after brief anaerobic maximal cycle ergometer exercise (P < 0.01).

Relationship between clinical features and binding domains of anti-prothrombin autoantibodies in patients with systemic lupus erythematosus and antiphospholipid syndrome.

Autoantibodies against prothrombin, including lupus anticoagulant antibodies (LAC), have been identified in patients with systemic lupus erythematosus (SLE) and antiphospholipid syndrome (APS). To identify the epitopes of LAC in patients with SLE and APS, we analyzed B cell epitopes of anti-prothrombin Abs. Prothrombin was purified from fresh plasma samples from healthy subjects, and fragmented by thrombin. Two fragments (prethrombin-1, 50 kDa, and fragment-1, 22 kDa) were separated and used for further experiments. The two fragments were coated on irradiated plate and the binding activities of sera from 13 patients with anti-prothrombin Abs (SLE, 7; APS, 4; SLE+APS, 2) were determined by using ELISA. The assay was conducted under the following conditions: use of irradiated plates, and TBS containing Tween-20. We detected two types of anti-prothrombin Abs. The first was anti-prethrombin-1 (n=5) while the other was Ab against fragment-1 (n=8). There were no patients with Abs that showed binding activities to both fragments. A higher proportion of patients with thrombosis were positive for anti-prethrombin-1 Abs (80%) than for anti-fragment-1 Abs (25%). Two patients with anti-prethrombin-1 Ab were positive for LAC and negative for anti-cardiolipin-beta2 glycoprotein I antibody (aCL-beta2GPI). Our results strongly support the notion that both prethrombin-1 and fragment-1 on prothrombin molecule are B cell epitopes.

Cervical adenocarcinoma, a novel combination chemotherapy with mitomycin C, etoposide, and cisplatin for advanced or recurrent disease.

We review the cases of 31 patients with stage IVb or recurrent cervical adenocarcinoma who were treated with combination chemotherapy utilizing mitomycin C, etoposide, and cisplatin (MEP). The total response rate was 16.1% (95% confidence intervals (CIs), 5. 5 to 33.7%) with 4 patients having a complete response (CR) and 1 having a partial response. In patients with no prior chemotherapy, the response rate was 26.7% (95% CIs, 7.8 to 55.1%) with 2 of these CR patients surviving over 3 years, 1 a disease-free survival. A marked response was found in distant recurrent lesions. The major toxicity was myelosuppression. Forty-five percent of patients had leukocytopenia above grade 3; thrombocytopenia and anemia were not common. In patients with cervical adenocarcinoma and no prior chemotherapy, there was a moderate response to MEP therapy.

Antimicrobial activity of Monascus pilosus IFO 4520 against contaminant of Koji.

Antimicrobial activity of Monascus pilosus IFO 4520 was examined to prevent contamination during beni-koji making in the open air. The antibacterial effect of the beni-koji prepared with this strain occured with 30 mg/ml of beni-koji extract in combination with 0.5% lactic acid against two contaminants of koji, Micrococcus varians and Bacillus subtilis. There were two compounds, antibacterial and antiyeast substances, in the beni-koji extract. These results suggest a possibility of inhibiting the growths of contaminants during beni-koji making using beni-koji extract and lactic acid.

Detergent and antigen fragility affect the ELISA for measurement of anti-prothrombin autoantibodies.

OBJECTIVE: Some investigators have reported that anti-prothrombin autoantibodies (aPT) in lupus anticoagulant positive sera were detectable by ELISA. Discrepancies in aPT ELISA were observed by some investigators. To clarify this situation, we tested the binding of aPT positive sera to purified prothrombin under various conditions. METHODS: We performed aPT ELISA under different conditions. The variables we tested were: ELISA plate (untreated or gamma irradiated polystyrene plates), buffer (phosphate buffered saline or Tris buffered saline), detergent (presence or absence of Tween-20), and antigen condition (intact or fragmented prothrombin). RESULTS: Anti-PT from patients with lupus or antiphospholipid syndrome were similarly bound to prothrombin with both buffers. Addition of Tween-20 to the buffer increased reactivity in the irradiated plate assay, but decreased reactivity in the untreated plate assay. Reactivities in 90% of lupus sera were decreased by the use of fragmented prothrombin. In contrast, the reactivity of serum from a healthy subject was remarkably increased by antigen fragmentation. CONCLUSION: The discrepant ELISA results in measurement of aPT in the various reports may have been due to the use of detergent in the buffer and condition of the prothrombin used as antigen. In our experiments the best ELISA condition for measurement of aPT was achieved using buffer with Tween-20 detergent, with prothrombin directly coated onto irradiated polystyrene plates.

[Early phase II trial of oral etoposide administered for 21 consecutive days in patients with cervical or ovarian cancer. ETP 21 Study Group--Cervical-Ovarian Cancer Group].

We conducted multi-site early phase II trial or oral etoposide administered for 21 consecutive days in patients with cervical or ovarian cancer in cooperation with 19 institutes. Fifty mg/body of oral etoposide was administered daily for 21 consecutive days. Cycles were repeated every 28 days. In cervical cancer, 24 patients were enrolled and 17 of them were evaluated. The overall response rate including CR and PR was 23.5% (4/17). In ovarian cancer, 18 patients out of 21 enrolled were evaluated. The overall response rate was 16.7% (3/18). The primary toxicity observed was myelosuppression such as leukopenia, neutropenia, hemoglobin decrease and thrombocytopenia. Other adverse effects were anorexia, nausea, vomitting, fatigue, alopecia and stomatitis. From these results we concluded that oral etoposide administered for 21 consecutive days was effective against cervical cancer.

[Prevalence of dementia and its subtypes in a rural area in Japan].

The prevalence of dementia in Japan in 1985 for people aged 65 years or older was 4.8%. Vascular dementia (VD) has been reported to be the commonest subtype A report from Tokyo in 1970 showed that it was 2.8 times more frequent than Alzheimer type dementia (SDAT). We assessed the prevalence of dementia in a rural area (population: 12,931, 25.3% were 65 years or older in 1994) in Kyoko. First, we questioned subjects about their demographic circumstances, memory disturbance, apraxia (agnosia) and daily activities. Our questionnaire was answered by 3.132 (95.8%) subjects, and 2,280 of them agreed to be examined by neurologists. Those who met the appropriate criteria of the DSM-III-R and NINCDS-ADRDA were diagnosed with dementia. By use of the Hachinski ischemic score (HIS), we distinguished VD from non-vascular dementia. In this study patients with non-vascular dementia were diagnosed with SDAT by the neurologists. Analysis of the data revealed that 4.8% of the study population was demented. According to the HIS results, only 3 of 15 had vascular dementia. The prevalence of dementia was the same as the average prevalence in Japan even though the elderly population of this town was twice as high as the average. It is hard to determine the prevalence of dementia in any community. Many factors must be taken into consideration: the coverage rate, the criteria for dementia, and whether to include institutionalized residents. The prevalence of dementia will increase with the aging of the population, and we must collect accurate data in order to plan efficiently.

[A case of stimulus-sensitive spinal myoclonus].

A 64-year-old man was admitted to our hospital because of a 2-year progression of muscular jerks on the bilateral thoracic and abdominal walls. Neurological examination was normal except for myoclonus localized in the bilateral Th5 to L1 segments. Myoclonus in this case was symmetrical and synchronous, and had a tendency to continue rhythmically. It was also segmental and considered to be spinal in origin. The interval between two adjacent myoclonic jerks was short when the amplitude and duration of the first myoclonus were high and long, respectively. We postulated that the activity of anterior horn cells at an impaired segment might be a factor that influences the resting time between jerks. A myoclonic jerk could be induced by a tendon tap given anywhere on this patient's body, with a latency ranging from 40 to 220 msec, irrespective of the sites of tapping. A long latency and polysynaptic reflex may participate in such stimulus sensitivity. The site of the reflex was probably above the motor nucleus of the trigeminal nerve, because jerks could be induced by jaw taps. Our observations support the theory that a spino-bulbo-spinal reflex is manifested at the impaired segment.

Expression of Xa1, a bacterial blight-resistance gene in rice, is induced by bacterial inoculation.

The Xa1 gene in rice confers resistance to Japanese race 1 of Xanthomonas oryzae pv. oryzae, the causal pathogen of bacterial blight (BB). We isolated the Xa1 gene by a map-based cloning strategy. The deduced amino acid sequence of the Xa1 gene product contains nucleotide binding sites (NBS) and a new type of leucine-rich repeats (LRR); thus, Xa1 is a member of the NBS-LRR class of plant disease-resistance genes, but quite different from Xa21, another BB-resistance gene isolated from rice. Interestingly, Xa1 gene expression was induced on inoculation with a bacterial pathogen and wound, unlike other isolated resistance genes in plants, which show constitutive expression. The induced expression may be involved in enhancement of resistance against the pathogen.