Photosynthetic traits of Siebold's beech seedlings in changing light conditions by removal of shading trees under elevated CO2.

The purpose of this study was to obtain basic information on acclimation capacity of photosynthesis in Siebold's beech seedlings to increasing light intensity under future elevated CO2 conditions. We monitored leaf photosynthetic traits of these seedlings in changing light conditions (before removal of shade trees, the year after removal of shade trees and after acclimation to open conditions) in a 10-year free air CO2 enrichment experiment in northern Japan. Elevated CO2 did not affect photosynthetic traits such as leaf mass per area, nitrogen content and biochemical photosynthetic capacity of chloroplasts (i.e. maximum rate of carboxylation and maximum rate of electron transport) before removal of the shade trees and after acclimation to open conditions; in fact, a higher net photosynthetic rate was maintained under elevated CO2 . However, in the year after removal of the shade trees, there was no increase in photosynthesis rate under elevated CO2 conditions. This was not due to photoinhibition. In ambient CO2 conditions, leaf mass per area and nitrogen content were higher in the year after removal of shade trees than before, whereas there was no increase under elevated CO2 conditions. These results indicate that elevated CO2 delays the acclimation of photosynthetic traits of Siebold's beech seedlings to increasing light intensity.

Examination of accuracy of sleep stages by means of an automatic sleep analysis system 'Sleep Ukiha'.

We examined the differences between the results of an automatic sleep analysis system and inspection decision. Subjects were 10 males (average age 21.6 years). One section consists of 20 s records. The sections that deviated from the algorithm could not be decided. Each sleep stage decided by automatic analysis was compared with the inspection decision. The agreement ratio of stage 3 was 91.6% in the highest, and followed by stage 2, stage 4, stage W and stage 1. The lowest was 62.5% for movement time. The total agreement ratio was 85.8%. The agreement ratios of the apnea index (AI) and the apnea hypopnea index (AHI) were relatively high, but for types of sleep apnea, agreement ratios require improvement.

Study on withdrawal of hypnotics: questionnaire on hypnotic use and its withdrawal.

To investigate the situation and problems contingent to hypnotic use and withdrawal, we conducted a questionnaire of outpatients. Only 41% of the patients were satisfied with their sleep and 53% of the patients took hypnotics. As regards the period, 83% of users had used them for more than 1 year and 19% had used them for more than 10 years. Although 90% of patients perceived efficacy of hypnotics, 67% felt more or less anxious about hypnotic use. Sixty-seven per cent of patients had actually withdrawn from the drugs or decreased dosage before. More than half the patients' conditions worsened after the withdrawal or reducing dosage.

Questionnaire relating to sleep paralysis.

A sleep survey was conducted on 8162 citizens. The cumulative experience rate of sleep paralysis was 39.6%. The initial occurrence of sleep paralysis peaked at age 16 years. In addition to being higher in young people than in older subjects, the incidence of sleep paralysis was also higher among women than among men, and was significantly higher among shift worker than non-shift worker, and among persons engaged in the nursing profession than those not engaged in the nursing profession. The experience rate of sleep paralysis demonstrated a strong correlation with the frequency of dreaming, the experience rate of nightmares, times and regularity of going to bed and waking up, and particularly with the degree of insomnia.

Hypnotic action of melatonin during daytime administration and its comparison with triazolam.

The present study was conducted to assess hypnotic action, effects on rectal temperature and dose dependency by daytime administration of exogenous melatonin (MLT) at 1 mg, 3 mg or 6 mg to subjects consisting of seven healthy juvenile adults. As a result, exogenous MLT significantly increased total sleep time and sleep efficiency, and MLT 6 mg was observed to demonstrate hypnotic effects that were nearly equal to those of triazolam at 0.125 mg. Rectal temperature was significantly decreased at MLT 1mg and 3 mg, there were no significant differences observed in the hypothermic effects at MLT 6 mg. These results indicate that exogenous MLT had dose-dependent hypnotic action on daytime sleep, and it is possible to consider that this hypnotic action was based on a direct-acting mechanism.

Purification, characterization, and application of a novel dye-linked L-proline dehydrogenase from a hyperthermophilic archaeon, Thermococcus profundus.

The distribution of dye-linked L-amino acid dehydrogenases was investigated in several hyperthermophiles, and the activity of dye-linked L-proline dehydrogenase (dye-L-proDH, L-proline:acceptor oxidoreductase) was found in the crude extract of some Thermococcales strains. The enzyme was purified to homogeneity from a hyperthermophilic archaeon, Thermococcus profundus DSM 9503, which exhibited the highest specific activity in the crude extract. The molecular mass of the enzyme was about 160 kDa, and the enzyme consisted of heterotetrameric subunits (alpha(2) beta(2)) with two different molecular masses of about 50 and 40 kDa. The N-terminal amino acid sequences of the alpha-subunit (50-kDa subunit) and the beta-subunit (40-kDa subunit) were MRLTEHPILDFSERRGRKVTIHF and XRSEAKTVIIGGGIIGLSIAYNLAK, respectively. Dye-L-proDH was extraordinarily stable among the dye-linked dehydrogenases under various conditions: the enzyme retained its full activity upon incubation at 70 degrees C for 10 min, and ca. 40% of the activity still remained after heating at 80 degrees C for 120 min. The enzyme did not lose the activity upon incubation over a wide range of pHs from 4.0 to 10.0 at 50 degrees C for 10 min. The enzyme exclusively catalyzed L-proline dehydrogenation using 2,6-dichloroindophenol (Cl2Ind) as an electron acceptor. The Michaelis constants for L-proline and Cl2Ind were determined to be 2.05 and 0.073 mM, respectively. The reaction product was identified as Delta(1)-pyrroline-5-carboxylate by thin-layer chromatography. The prosthetic group of the enzyme was identified as flavin adenine dinucleotide by high-pressure liquid chromatography. In addition, the simple and specific determination of L-proline at concentrations from 0.10 to 2.5 mM using the stable dye-L-proDH was achieved.

Development and application of the 'Sleep Ukiha' automated sleep analysis system.

We attempted to develop an automated sleep analysis system that uses a personal computer as an aid to the entire sleep research process. Analysis is based on electroencephalogram, chin muscle electromyogram and electrooculography, while other physiological phenomena can be handled arbitrarily. Major characteristics of the system include: (i) simultaneous gathering of physiological phenomena from up to three patients; (ii) high-speed waveform analysis; (iii) user-friendly operating environment through the use of a graphical user interface; and (iv) versatile utilization of analytical data in research.

A narcoleptic patient exhibiting hallucinations and delusion.

A female narcoleptic patient with persistent auditory hallucinations and delusion is reported. These psychotic symptoms appeared before the diagnosis of narcolepsy and before the use of pemoline and clomipramine. On admission to the Department of Neuropsychiatry at the Kurume University school of Medicine, the patient's auditory hallucinations were almost the same as the previous hallucinations. The patient's delusional state improved smoothly after haloperidol administration. After that, the patient related well socially. The persistent auditory hallucinations were correlated with rapid eye movement sleep symptoms during her disease process so that we concluded that her persistent auditory hallucinations and delusion of observation were due to narcoleptic symptoms.

An experimental transplantable osteosarcoma with spontaneous pulmonary metastasis in hamsters.

Animal models of osteosarcoma with spontaneous pulmonary metastasis which retain metastatic capacity and osteoid formation after serial passages have been reported infrequently. In this communication we describe some biological features of a transplantable osteosarcoma, Os515, induced by BK-virus in Syrian golden hamsters. The subcutaneously transplanted tumours in 2-week-old animals grew progressively until death, with a mean survival time of 32 days. Distant metastases occurred only in the lungs in all animals. The histological appearance was osteosarcoma of osteoblastic type. Enzyme-histochemical staining showed alkaline phosphatase activity in many cells and beta-glucuronidase activity in few cells. Tumours transplanted intramuscularly in the hind limbs were amputated radically at 5 or 11 days. A small number of animals died from lung metastases without local relapse during the observation period of 140 days after grafting. All the control hamsters bearing unamputated tumours died much earlier. Necropsy revealed large metastatic nodules in the lungs of limb-amputated animals and small diffuse nodules in the lungs of untreated control animals. The development of lung metastases was monitored by soft X-ray without sacrificing the animals. This model will be useful in studies of mechanisms of metastasis and for the experimental treatment of osteosarcoma.

[Biological characterization of a hamster osteosarcoma Os 515--development of lung metastasis].

Biological characterization of a transplantable hamster osteosarcoma Os515 which had been induced by BK virus was attempted, and usefullness of this tumor was confirmed as an experimental model of treatment of osteosarcoma. All the hamsters with this tumor died of spontaneous lung metastasis. The chest X-ray picture of a tumor-bearing hamster visualized diffuse metastasis in the whole lungs 30 days after transplantation. The development of lung metastases was monitored by chest X-ray after marginal excision of subcutaneously transplanted tumors or after radical disarticulation of a tumor-bearing hind limb. Slow-growing nodular lung metastases were seen on X-ray pictures when the subcutaneous tumors were resected 1 or 2 weeks after transplantation, whereas fast-growing diffuse metastases were seen when the tumors were not resected at all or resected 3 weeks after transplantation. Nodular lung metastases occurred in a small number of animals long after the early radical disarticulation of a tumor-bearing leg.

[Combined therapy with microwave hyperthermia and adriamycin in MM2-bearing mice].

C3H/He mice were inoculated i.p. with synergic MM2 tumor cells(2 X 10(6) cells/mouse), and subsequently treated either by systemic hyperthermia induced by microwave irradiation (2450 MHz)alone, administration of ADM alone, or a combination of both. The mice were exposed to hyperthermia for 5 minutes everyday at an output of 10, 20 or 30 watts. ADM was administered i.p. for 3 days at dose levels of 0.025 mg/kg, 0.05 mg/kg or 0.075 mg/kg body weight. The mean survival time and the mean relative body weight were recorded. From these data, tumor-suppressing effects of the treatments were analysed by one-way or two-way variance analysis. Mice treated with 30-watt microwave irradiation survived significantly longer than control mice without treatment (P less than 0.05), as did mice treated with ADM alone (P less than 0.01). The mean relative body weight was similar among the microwave, ADM and control groups. When the mice were given microwave treatment combined with ADM, significant difference was observed in the mean survival time by one-way variance analysis (P less than 0.01), but no significant difference was observed by two-way variance analysis. The best condition of mice and the optimal dose of ADM still remain to be determined to facilitate more efficient tumor-suppression using this combination therapy.

[Tumor suppressing effect of systemic hyperthermia with 2350 MHz microwaves combined with 5-fluorouracil in mice].

C3H/He mice were inoculated i.p. with syngeneic MM2 tumor cells (2 X 10(6) cells/mouse), and they were subsequently treated by systemic hyperthermia with microwave irradiation (2450 MHz) alone, administration of 5-FU alone, or a combination of the both. The mice were exposed to hyperthermia for 5 minutes everyday with an output of 10, 20 or 30 watts. 5-FU was administered i.p. for 3 days at dose levels of 2.5 mg/kg, 5 mg/kg or 10 mg/kg body weight. The mean survival time and the mean relative body weight were recorded. From these data, tumor-suppressing effects of the treatments were analyzed by one-way variance analysis or two-way variance analysis. The mice treated with microwave irradiation alone or 5-FU alone survived significantly longer than the control mice receiving no treatment (P less than 0.01). However, no significant difference in mean survival time was observed between the microwave group and the 5-FU group. The mean relative body weight was similar among the microwave, the 5-FU and the control group. When the mice were treated with microwave irradiation and 5-FU, significant difference was observed in the mean survival time (P less than 0.01), and the combination of microwave irradiation with an output of 20 W and 10 mg/kg of 5-FU seemed to be optimal. Determination of an optimal combination was difficult with the mean relative body weight, because body weight decreased due to side effect of the combination therapy.

[Experimental study on the anti-tumor effect of microwaves].

C3H/He mice bearing a mammary carcinoma, MM2, were treated by systemic hyperthermia with microwave irradiation (2450 MHz). An output of 30 watts of microwaves for 5 min at a distance of 20 cm from the animal raised and maintained the rectal temperature of mice to higher than 42 degrees C for 2.2 min and higher than 40 degrees C for 10.2 min. The mice were inoculated i. p. with MM2 tumor cells (2 X 10(6) cells/mouse), and then they were exposed to hyperthermia every day; 10 watts for 2, 4, or 6 min, or 20 watts for 2, 4, 6 min. The control mice receiving no irradiation and the mice treated with irradiation of 10 watts for 2 min or 20 watts for 2 min died within 21-23 days. Whereas, 40 percent of the mice treated with microwave irradiation of 10 watts for 4 or 6 min, or 20 watts for 4 or 6 min survived longer than 30 days. The 6 mice, which survived longer than 120 days, were challenged by reinoculation with MM2 tumor cells (2 X 10(6)cells/mouse). All the mice but one survived longer than 30 days with no accumulation of ascites. These experimental data suggest that the systemic hyperthermia with microwave irradiation (24 50 MHz) might suppress the tumor growth in vivo.