Electrical stimulation with twice the threshold intensity and a duration of 10 msec was given to an isolated rat papillary muscle at a frequency of 1 Hz, and the influences of diltiazem, verapamil, papaverine, isoproterenol, adrenaline and noradrenaline were examined on contractile tension, contraction velocity and relaxation velocity of the muscle.
1) When the muscle was kept in a modified Locke's solution in which the CaCl2 was reduced from 1.6 to 0.8 mM, i. e., in a low Ca²⁺ medium, each of contractile tension, contraction velocity and relaxation velocity decreased: the degree of the decrease in contractile tension was greatest, that in contraction velocity was second and that in relaxation velocity was smallest. When the muscle was kept in a high Ca²⁺ medium in which the CaCl2 was contained in a concentration of 3.2 or 6.4 mM, each of the tension and velocities increased: the greatests were those of contraction and relaxation velocities and the smallest was that of contractile tension.
2) Each o f isoproterenol (0.01 to 0.1 mg/l), adrenaline (0.1 to 10 mg/l) or noradrenaline (0.1 to 6.0 mg/l) increased contractile tension, contraction velocity and relaxation velocity: the increase in contraction and relaxation velocity was remarked. The effect of the drugs on the muscle which was kept in the low Ca²⁺ medium was less than that on the muscle kept in a standard Locke's solution, and the effect was scarcely increased, also, even when the muscle was kept in the high Ca²⁺ medium.
3) Diltiazem (0.1 to 1.0 mg/l) or verapamil (0.001 to 0.1 mg/l) decreased each of contractile tension, contraction velocity and relaxation velocity of the muscle: the mode of the inhibitory action of the drugs resembled each change of them which was seen when the muscle was kept in the low Ca²⁺ medium without any drug. In the presence of calcium antagonists, the addition of CaCl₂ to the medium caused a more remarkable increase in contractile tension, contraction velocity and relaxation velocity than the addition of isoproterenol.
4 ) Papaverine (0.1 mg/l) decreased contractile tension of the muscle, while increased the contraction and relaxation velocities. However, the drug (10 mg/l) increased all of them: the increase in the contraction and relaxation velocities resembled the increase induced by the high Ca²⁺ medium without any drug.
5) The increase in contractile tension, contraction velocity and relaxation velocity induced by the high Ca²⁺ medium was suppressed by the presence of papaverine (10 mg/l), and the increase in contractile tension induced by isoproterenol (0.01 mg/l), also, was suppressed by the presence of papaverine (0.01 mg/l).
From the results, it is presumed that there are two kinds of Ca²⁺ channels in the plasma membrane of papillary muscle: one is a channel which is voltage dependent and the other is a channel which is activated by Ca²⁺-calmodulin dependent proteinkinase (in an evironment in which the concentration of Ca²⁺ is excess) or by cyclic AMP dependent proteinkinase (in the presence of isoproterenol or another adrenergic,β-stimulant). The former voltage dependent channel is thought to be a main Ca²⁺ channel through sarcoplasmic membrane and to be modified by the latter channel activated by these proteinkinases. The contractile tension, contraction velocity and relaxation velocity are influenced by calcium ion concentration in the medium, while the velocities are considered to be influenced by cyclic AMP level which controls calcium concentration in an intracellular fluid.

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It is generally accepted that Rubella virus (RV) causes weak cytopathogenecity. Cytopathic effect (CPE) by RV has been rather irregular and difficult to recognize in tissue culture cells. Therefore, investigation of RV replication must be taken much effort.
In the present study, the author applied Enzyme focus forming a ssay (EFFA) which was developed for the titration of such weak cytopathogenic viruses and studied RV replication morphologically.
Serially diluted RV was inoculated on Vero cells which were cultured in multiwell plates. Cells were added mainteance medium containing 1.5% methylcellulose. After 7-10 days incubation at 37°C, the infectious foci were stained with either hoseradish peroxidase conjugated anti RV IgG, Fab' fraction (HRP-Fab'), or Giemsa's solution. Though the infectious foci were not shown by Giemsa staining, immunoperoxidase positive foci were stained clearly and it was easy to count the RV titer as EFFU. The immunoperoxidase positive foci were embeded as immunoelectron microscopic specimen. Following results were obtained in the present study.
1. RV, M-33 strain, did not cause clear CPE on Vero cells.
2. RV titer, measured by 50% tissue culture interference dose (TCInD₅₀), increased logarithmically from 2 to 8 days after infection. The TCInD50 of 8 and 10 days after in f e ction were 10^6-3 and 10^6-0 respectively and kept the same titer after 10 days. Instead o f these viral titers, clear CPE was not found. RV infeced Vero cells were resemble to the cel l s in which the other virus infected persistently.
3. RV specific fluorescence was observed around the nucleus of Vero cells 3 days after infection. The specific fluorescence spread from perinuclear area to cytoplasm by lineal y or spotted pattern according to the infections time has passed.
4. Similar findings were obtained when the cells were stained with HRP-Fab'.
5. Replicated RV formed infectious foci when assayed by EFFA. These foci were easily observed and caliculated macroscopically. Uniform foci,0.8 mm and 1.0 ^-1.2 mm in diameter, were formed on the plates of 7 days and 10 days after the inoculation respectivey. The titers of specimen 4 and 10 days after the infection were well correspond e d to the titers of T CInD₅₀. In addition, the titer of EFFA was obtained earlier than the tite r of TCInD₅₀.
6. Aggregates of electron dense particles,20-30 nm in diameter, were observed beneath the Vero cell membrane and cytoplasmic vacuoles in the specimen of 3 days after infection b y electron microscope. Mature RV particles were observed 4 days after infection. Further more, RV, budding from the cell membrane, were observed.
7. In the specimen of immunoenzyme electron micrscope (IEM), which was processed from the foci of EFFA, heavyly stained particles of 20-30 nm in diameter were observed at the peripheral area of the Vero cells. These particles corresponded to the particles found o n the conventioanl electron microscocopic study. Therefore, the electrondense particles foun d in the cells 3 days after RV infection were identified as the immature form of RV i n the cytoplasm.
8. In the IEM study, microvilli of RV infected Vero cells were stained heavyly. Furthermore, particles of 60-100 nm in diameter, outside of the microvilli were stained. T h ese particles were thought to be mature RV which budded out from the microvilli.
9. From these findings, the author conclude that the proteins which concerned RV were formed perinucleally in the Vero cells. RV precursor particles were most recongnized j ust underneath the cell membrane or vacuole membrane and they bud through these membra n e and form mature RV particles.

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As a part of the study to elucidate the influence of rhythmical sound on the central nervous system activity, the effect on the threshold of arousal reaction and evoked muscular discharge was investigated in the rabbit and the following observed.
1) The threshold of the arousal reaction, appearing in the cerebral cortex EEG and the hippocampus EEG and the muscular discharge in the fore- and hind limbs, due to stimulati on of the midbrain reticular formation (RF) with 100 Hz, was increased by the rhythmical sound of 60/min beat frequency and 500 Hz pitch.
2) The threshold of the arousal reac tion and the evoked muscular discharge was decreased by the rhythmical sound of 450/min beat frequency and 1000 Hz pitch.
3) The decrease or increase in the threshold of the arousal rea ction and evoked muscular discharge following sonic stimuli (500 Hz 60/min or 1000 Hz 450/min), was abolished by destruction of the nucleus ventralis anterior or nucleus reticularis, which belong to the diffuse thalamocortical projection system.
From these results, it w as concluded that excitability of the brain stem reticular formation is inhibited by rhythmical sound of a low frequency and low pitch (500 Hz 60/min) and facilitated by rhythmical sound of a high frequency and high pitch (1000 Hz 450/min). These findings suggest that the decrease in excitability of the RF by rhythmical sound of a low frequency and low pitch and the increase in excitability of the RF by rhythmical sound of a high freguency and high pitch can be attributed to a rise in indirect inhibition and a decrease in indirect inhibition of the RF, respectively, via the diffuse thalamocortical projection system.

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A quantitative understanding of dietary contents is undoubtedly important in hemodialysis patients. Until now no simple method to evaluate dietary contents has been available. We have developed a simple method to calculate the amounts of protein, sodium and potassium intake in hemodialysis patients, and compared Gotch's formula based on a urea kinetics model with our simple method.
[Method]
In 15 chronic dialysis patients in nutritionally steady state, urea clearance and the pre-, post-, dialytic body weight, BUN, serum [Na⁺] and [K⁺], and the same parameters bofore the next hemodialysis, were measured. From these values, the total body fluid volume (V) and urea generation rate (Gu) can be obtained based on urea kinetics (Gotch's formula).
On the other hand, V and Gu were also obtained by the simple method by assuming that dry weight is equal to normal V. From Gu (mg/mm), obtained either by means of Gotch' s formula ( I ) or the simple method ( I ), protein catabolic rate (PCR) can be estimated { (PCR (g/day) = (Gu +1.2) X 9.35}; and it was assumed that PCR is equal to the amount of protein intake. The amount of sodium intake was calculated as the change in the product of V and serum CNa⁺), by assuming that the absolute amount of intracellular effective osmotic cations was constant. The amount of potassium intake was calculated in the same manner.
[Results]
1. The postdialytic V and the amount of protein intake determined respectively by means of ( I ) and (II) were30.9 ± 1.6l (57 ± 2% body weight) and 32.2 ± 1.5/ (59 ± 2% body weight), and 66 ± 2g/day (1.3g/kg/day). and 69 ± 3g/day (1.3g/kg/day). Highly significant correlations were found between the two methods (p<0.001, p<0.001).
2. Highly significant correlations for the amounts of sodium and potassium intake were also found between the two methods (p<0.001, P<0.001). Sodium intake determined by methods ( I ) and (II) respectively was 334 ± 38mEq/interdialysis (187 ± 21mEq/day) and 328 ± 38mEq/interdialysis (183 ± 21mEq/day). Potassium intake was 73 ±4mEq/interdialysis (41 ± 2mEq/day), and 76 ± 4rnEq/interdialysis (42 ± 2mEq/day), respectively.
[Conclusion]
We have developed a simple method to calculate the amounts of protein, sodium, potassium intake in hemodialysis patients without carrying out a dietary investigation or measuring extracellular fluid volume and confirmed that it is useful. We believe that this method will prove to be a useful tool for assessing a nutritional state and examining the relation between protein intake and anemia or between sodium intake and blood pressure.

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Among the recent advances in cancer therapy, the development of chemotherapy is most remarkable. On the other hand, hemorrhagic diathesis is often caused by antineoplastic drugs, especially Anthracyclines, which are widely used in the treatment of various kinds of cancer.
Reported here are the results of serial experiments on the influence of antineoplastic drugs on human platelets in vitro. The effects of Coenzyme Q₁₀ (CoQ₁₀), known to alleviate such adverse actions as cardiotoxicity and to increase the antineoplastic efficacy of Anthracyclines, were also investigated. The resu lts were as follows:
(1) Anthracyclines, including Adrimycin (ADM), Daunomycin (DM), and Aclacinomycin A (ACR), inhbited platelet aggregation and release reaction in dose and time-dependen t manner.5-Fluorouracil (5-FU), a non-Anthracycline, did not show su c h inhibitory effects.
(2) Differences were found in the intensity of the action between Anthracyclines, of which DM was the most intensive regardless of the aggregating agent. ADM inhibited pl a t elet aggregation more effectively than ACR when ADP, collagen, epinephrine, and A 23187were employed as agonists. ACR was more effective than ADM in the case of thro m bin. Anthracyclines had almost no inhibitory effect on arachidonic acid and STA₂ induced platelet aggregation.
(3) CoQ₁₀ also had an inhibitory effect on platelet aggregation, which was more remarkable in the first phase of aggregation. Furthermore CoQ₁₀ was shown to enhance the a n tiplatelet effect of ADM.
(4) Neither Anthracyclines nor CoQ₁₀ caused any significant change in cyclooxygenase metabolites or cyclic nucleotide levels, which might account for their inhibitory effec t s on platelet function.
(5) The liberation of radioactivity from ¹⁴C-arachidonic acid-labeled platelets in response to collagen was reduced by Anthracyclines. It was suggested that the impairment of platele t function by Anthracyclines was partially attributable to the decrease of phospho l ipase activity.

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An electrophysiological study was made with kindled rabbits (which were experimentaly produced in the amygdaloid nucleus, hippocampus, septal nucleus and cerebral cortex) on the electrical changes of photopalpebral reflex (PPR) and photo-evoked eyelid microvibration (MV).
The results obtained were as follows.
1) In control group, no rema r kable change was observed in the PPR and MV.
2) In amygdaloid kindled-group, the period required for the completion of the kindling effect was 7 to 21 days (average,16 days). The afterdischarge was propagated to cerebral cortex and hippocampus on the first trial.
3) In amygdaloid-kindled group, the afterdischarge threshold was from 0.7 upto 2.6mA (average,1.5mA). The duration time of afterdischarge extended from 2 to 13 sec. (average,6sec. ) on the first trial to 12 to 86 sec. (average,46.8 sec. ) on the last trial after the kindling effect was completed.
4) In amygdaloid -kindled group, the amplitude of MV increased in the early stage, but stepped down later. The amplitude of PPR_{1_4,6,9} showed no marked change. The PPR_7,8increased remarkably and the PPR5 slightly following the initiation of stmulation.
5) In hippocampus-kindled group, the period required for the comp letion of the kindling effect was 19 to 190 days (average,72 days). The afterdischarge was propagated to the amygdaloid nucreus, but not to the cerebral cortex on the first triral.
6) In hippocampus-kindled group, the afterdischarge threshold was from O.6 upto 1.4 mA (average,1. imA). The duration of the afterdischarge extended from 6.5 to 16 sec. (average,9.3 sec. ) on the first trial to 12 to 281 sec. (average,178 sec) on the last trial.
7) In hippocampns-kindled group, the amplitude of MV increased remarkabl y following the initiation of stimulation. The amplitude of PPR _1-4 showed no marked change, while the PPR_5-8 increased in amplitude. The PPR₉ either increased or decreased by cases.
8) In septal nudes-kindled group, the period required for the completion of the kindling effect was 17 to 38 days (average,26 days). The afterdischarge was propagated to the amygdaloid nucleus and hippocampus, but not to the cerebral cortex except one case on the first trial.
9) In septal nucleus-kindled group, the afterdischarge threshold was from 1.8 upto 4.6mA (average,3.4mA). The duration of the afterdischarge extanded from 2 to 16 sec. (average,6.8 sec. ) on the first trial to 20 to 311 sec. (average,121 sec. ) on the last trial.
10) In septal nucleus-kindled group, the amplitude of PPR_1-4 showed no mark ed change, while the PPR_5-9 incresed. The amplitude of MV increased in the early stage and later decreased gradually.
11) In ce rebral cortex-kindled group, the period required for the completion of the kindling effect was 16 to 64 days (avetage,33 days). The afterdischarge was propagated to the amygdaloid nucleus and the hippocampus on the first trial.
12) In cerebral cortex-kindled group, the afterdischar ge threshold was from 0 4 upto 3.8mA (average,1.4mA). The duration of the afterdischarge extended from 5 to 28sec. (average,13.6 sec. ) on the first trial to 14 to 206 sec. (average,67.6sec. ) on the last trial.
13) In cerebral cortex-kindled group, the amplitude of MV increased. The amplitude of PPR_1-4 showed no marked change, while the PPR_5-9 increased.

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Psychogenic amnesia is known as psychogenic loss of personal identity, fugue, change of personal identity and retorograde amnesia. The cases, in which patients complained of a complete loss of memory for personal identity as well as whole life, have been reported and known as psychogenic total amnesia.
In this communication, the present author has reported two cases of psychogenic amnesia, of whom one is a 28-year-old male who complained of a complete loss of memorry for his previous life. The other is a 20-year-old female patient with Ganser' s syndrome due to dissociative reaction.
The author has focused upon thier depressive states in this discussion. In these cases, there were scarcely comfortable and agreeable families for them, moreover they were not enough mature to deal with emotinal conflicts of actual life. They had become depressive and lonsome, then these cases had to lose memory for thier personal identity.
There are a number of papers, where the above-mentione d depressive moods were discribed. Generaly speaking, psychogenic amnesia are widely accepted as hysterical dissociative reaction.
Amnesia can be recognized as a defence mechanism of the depressive states. Based on the own experience, the present author has concluded that psychogenic amnesia should be considered in a relationship to depression.

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