[Clinical biochemical evaluation of central fatigue with 24-hour continuous exercise].

OBJECTIVE: The present study was conducted to clarify the effects of ultra-marathon (ultra long-term aerobic exercise in which people run long distances) on the brain; examine the issue of central fatigue; verify the serotonin hypothesis of exercise-induced brain fatigue, and ascertain relationships between central fatigue and oxidative stress. METHODS: Subjects consisted of 15 individuals (12 men, 3 women) who ran continuously for 24 h. Mean age was 44 +/- 9 years (range, 31 approximately 64 years). Blood tests were conducted: (1) before starting to run (around 09:00); (2) 16h after starting (02:00 the next day); and (3) just after the finish (around 10:00 the next day) to measure the serum levels of serotonin, melatonin, free tryptophan (f-Tp) and free fatty acid. At the same time, urine samples were collected to measure levels of urinary biopyrrins (BPn). Subjective symptoms were investigated using the Japanese version of the Profile of Mood States (POMS) instrument. RESULTS: (1) Participants ran a mean (+/- SD) distance of 162.6 +/- 18.3 km. (2) There were not marked changes in serum serotonin levels. Serum melatonin levels at 3 time points were 3.4 +/- 0.6 pg/ml, 57.2 +/- 15.2pg/ml and 7.8 +/- 8.9pg/ml, respectively(p < 0.01 before start vs. 16h after start). Serum f-Trp levels at the 3 time points were 5.4 +/- 0.9 nmol/ml, 9.7 +/- 2.1 nmol/ml and 11.5 +/- 4.9 nmol/ml, respectively (p< 0.05 before start vs. just before finish). Free fatty acid levels were 0.42 +/- 0.10 nmol/ml, 1.26 +/- 0.11 nmol/ml and 1.39 +/- 0.23 nmol/ml, respectively (p < 0.01 before start vs. 16 hours after start) (p < 0.05 before start vs. just after finish). (3) Urinary BPn levels increased with time, from 1.2 +/- 0.7 nmol/ml to 2.6 +/- 1.0 nmol/ml to 4.0 +/- 1.5 nmol/ml, respectively (p < 0.01 before the start vs. 16 hours after the start). (4) In terms of POMS scores, fatigue score (Factor F) increased, but vitality score (Factor V) was high at all time points and did not demonstrate any marked changes. Scores for anger and hostility were low (Iceberg profile-type: convex type). Urinary BPn levels were correlated significantly with both serum f-Trp level and Factor F:(y = 8.41x + 2.5, r = 0.708, n = 42) and (y = 2.82x + 5.9, r = 0.568, n = 42), respectively. Urinary BPn thus reflected the degree of subjective fatigue with a high level of sensitivity. CONCLUSIONS: The present results suggest that running continuously for 24h induces brain fatigue and that oxidative stress may be involved.

[Current situation of the malaria inspection in Jikei University Hospital].

The current situation of the malaria inspection in our laboratory was investigated. Malaria was detected by three different methods, May Giemsa staining(MG), acridine orange staining(AO), and antigen detecting method using NOW ICT Malaria P.f./P.v. kit(Ag). There were 207 requests a year(17.3 per month), and the holiday/night request occupied 12%. Fifteen patients were positive, 5 with plasmodium falciparum (p.f.) and 10 with plasmodium vivax(p.v.), including 3 relapsed cases. All the patients with p.f. were suffered in Africa, and 6 with p.v. were in Southeast Asia, and one with p.v. was in Central America. The rate of coincidence between MG/Ag and MG/AO were 94.4% and 96.9%, respectively. There were 7 samples that were MG negative and Ag positive, but all of these samples were obtained after the initiation of the treatment. There was no sample that showed MG positive and Ag negative. Our data suggested that no difference in detection sensitivity was found between microscopic observation and the antigen detection kit. Thus it would be a very useful and accurate strategy to use this antigen detection kit in a routine laboratory check up.

A novel positive regulatory element for exfoliative toxin A gene expression in Staphylococcus aureus.

A 1.4 kb positive regulatory element (ETA(exp)) that controls staphylococcal exfoliative toxin A (sETA) transcription was cloned from Staphylococcus aureus. ETA(exp) is located upstream of the cloned 5.8 kb eta gene (etaJ1) obtained from the chomosomal DNA of S. aureus ZM, the standard ETA-producing strain. The cETA prepared from an Escherichia coli transformant into which the recombinant plasmid petaJ1 (5.8 kb eta/pUC9) had been introduced was expressed at high levels in the culture supernatant and the ammonium-sulfate-precipitated culture supernatant fraction as shown by immunoblotting and the single radial immunodiffusion test. However, cETA produced by the recombinant plasmid petaJ3 containing the 1.7 kb eta sequence (etaJ3) with a 1.45 kb ETA(exp)-deficient eta fragment (1.7 kb eta/pUC9) obtained from the 5.8 kb eta sequence by subcloning was not detected in either the culture supernatant or the ammonium-sulfate-precipitated culture supernatant fraction (167-fold concentrate of the culture supernatant) by immunoblotting or the single radial immunodiffusion test. A large amount of cETA was produced by the 1.7 kb eta sequence when it was linked to ETA(exp) amplified by PCR (1.7 kb eta-ETA(exp)/pUC9), regardless of the orientation of ETA(exp) insertion. Northern blot hybridization showed lower levels of the transcripts of the 1.7 kb eta sequence than of the 5.8 kb eta sequence. The rsETA prepared from an S. aureus transformant into which the recombinant plasmid 3.4 kb eta-ETA(exp)/pYT3 (pYT3-etaJ6) had been introduced was expressed at high levels in the culture supernatant fraction as shown by the latex agglutination test. However, the agglutination titre in the culture supernatant fraction of rsETA produced by the recombinant plasmid (1.7 kb eta/pYT3) containing the 1.7 kb eta sequence carrying the 1.4 kb ETA(exp)-deficient eta fragment (pYT3-etaJ3) was 2500-4000 times lower than that of pYT3-etaJ6.

[Evaluation of the simultaneous detection system for Chlamydia trachomatis/Neisseria gonorrhoeae DNA by the isothermal and chimeric primer-initiated amplification of nucleic acids (ICAN)].

The isothermal and chimeric primer-initiated amplification of nucleic acids (ICAN) is a new isothermal DNA amplification method composed of exo Bca DNA polymerase, RNaseH and DNA-RNA chimeric primers. We developed the simultaneous detection system for Chlamydia trachomatis/Neisseria gonorrhoeae DNA, combined with luminescence detection by a probe hybridization. In the performance tests, this system was able to detect 10 to 100 copies of C. trachomatis/N. gonorrhoeae DNA for only 3.5 hours, and was highly specific to C. trachomatis/N. gonorrhoeae without any cross-reaction to C. pneumoniae, N. lactamica, N. sicca or N. meningitidis. When we tested 60 clinical samples of urine and cervical swabs, the interpretive results were completely consistent with those obtained by Roche PCR system. Of 13 positive samples by the ICAN and PCR systems for C. trachomatis, four were negative by EIA method(IDEIA Chlamydia). These results indicate that the ICAN system is an efficient and sensitive system to simultaneously detect C. trachomatis/N. gonorrhoeae DNA.

Apnea-related heart rate variability in congestive heart failure patients.

Sleep-disordered breathing (SDB) causes fluctuation of the RR interval. However, the details are uncertain. We studied the characteristics of sleep-related heart rate variation (HRV) in congestive heart failure (CHF) patients with SDB. Ambulatory electrocardiograms and data on respiration (oronasal flow, trachea sound, abdominal wall movement, and oxygen saturation) were simultaneously recorded by a multi-channel digital recorder for 13 CHF patients (8 men and 5 women; mean age, 68 +/- 4 years). Heart rate variation occurred as a result of cyclical apnea attacks between 0.005 and 0.03 Hz (apnea band). The proportion of the apnea band (% apnea) increased with the number of apnea episodes, and SDB was highly likely when the % apnea was > or = 80%. Low-flow oxygen administration effectively reduced apnea frequency, and the apnea-related HRV also decreased. We concluded that apnea-related HRV was useful for detecting and following SDB in CHF cases.

Apnea-related heart rate variability and its clinical utility in congestive heart failure outpatients.

BACKGROUND: Cheyne-Stokes breathing (CSB) is an abnormal cyclical pattern of respiratory fluctuations observed during sleep in congestive heart failure (CHF) of poor prognosis. We examined the clinical usefulness of CSB screening using the heart rate variability (HRV) data from the ambulatory electrocardiogram. METHODS: We monitored ambulatory electrocardiograms and respiration simultaneously in 86 heart disease patients of both sexes, aged 57 +/- 1 years. HRV was analyzed by the maximum entropy method during the sleeping period (11 PM-5 AM). The 43 CHF patients underwent a 1-year follow-up study. RESULTS: In the power spectra of the HRV, peaks were observed within the CSB band (0.005 to 0.03 Hz). Statistically significant differences in HRV were observed between CSB patients and CSB-free patients in very low frequency (VLF) (P = 0.04), VLF/total frequency (TF) (P = 0.02), CSB (P = 0.01), CSB/TF (P = 0.003), and CSB/VLF (P < 0.0001). Cardiac events occurred in 23% of patients, including cardiac death in two, and rehospitalization for aggravated CHF in eight. In a multivariate Cox regression analysis in which age, sex, ejection fraction, NYHA functional class, beta blocker use, and basic heart disease were included, absence of ACE inhibitor use (RR 5.5, 95% CI 1.0-31) and CSB/VLF > or =80% (RR 4.2, 95% CI 1.1-17) remained significant predictors of cardiac events. CONCLUSIONS: HRV can act as an indicator of the presence of CSB in CHF patients, and could therefore be used, under outpatient conditions, to identify a CHF patients with a poor prognosis.

[Bacillus anthracis, prions, Coxiella burnetii].

The detection kits for Bacillus anthracis, an isoform of host prion and Coxiella burnetii, using genetic technology are still not generalized. For B. anthracis target genes for detection would be genes of toxins. Although it is difficult to detect prions related to prion diseases before death, some mutants of prion gene in leukocytes and 14-3-3 proteins in cerebrospinal fluid are detectable in living patients suffering from Creutzfeldt-Jakob disease. The gene of superoxide dismutase in Coxiella burnetii is a useful target for detecting this Rickettsiae.

Oxygen desaturation and heart rate variability due to Cheyne-Stokes respiration in congestive heart failure patients.

Cheyne-Stokes respiration is common in congestive heart failure patients and causes cyclical fluctuation of the RR interval. We studied the characteristics of apnea-related heart rate variability (apnea HRV) and the relation between apnea HRV and oxygen desaturation was examined. Ambulatory electrocardiograms and data on respiration (oronasal flow, tracheal sounds, abdominal wall movement and oxygen saturation) were simultaneously recorded by a multi-channel digital recorder for 16 congestive heart failure patients (10 men and 6 women; mean age, 68 +/- 13 years). HRV occurred as a result of cyclical apnea attacks between 0.005 and 0.03 Hz (apnea band). Apnea HRV was obtained as the power ratio of apnea HRV normalized by the very low frequency band (0.003-0.04 Hz). Apnea HRV increased with the number of apnea episodes and the oxygen desaturation index, but no relation between apnea HRV and either mean oxygen density or oxygen desaturation time was observed. We concluded that apnea HRV is a predictor of the number of apnea attacks or oxygen desaturation, but does not reflect the degree of oxygen desaturation in Cheyne-Stokes respiration.