Culex pipiensvar.molestusの卵巣組織をOrigmとした株化細胞に日本脳炎ウイルス(JEV)のマウス脳継代馴化株JaGAr-01株と,蚊より分離された新鮮野生株JaOAr-100株,および,JaOAr-121株を感染させ,次の成績を得た.
1.何れのJEV株とも,細胞変性効果を伴なって増殖し,感染4～5日後には,ウイルス量も10^6～7PFU/mlとなった.なお,JaGAr-01株は,他のウイルス株に較ベ速やかに増殖し,細胞変性効果の出現も早期に認められた.また,JaOAr-121株では,細胞融合の傾向が強く認められた.
2.何れのJEV株とも,持続感染が証明された.即ち,感染3箇月後には10^5～6PFU/mlのウイルスを培養液中に認め,また,16箇月後においても,なおウイルスを証明する事が出来た.
3.何れのJEV株とも,感染の経過が進むに従い,非感染細胞に比較して,Acid phosphatase活牲の顕著な増加が,形態学的にも,定量的にも認められた.また,Acid phosphatase活性の上昇は,細胞変性効果に先行して証明出来た.
4.何れのJEV株とも,感染16～20時聞後に,抗JaGAr-01株免疫家兎血清のγ-Gとの間に,螢光抗体法による特異螢光を核膜周辺部に認めた.
5.感染細胞の電子顕微鏡による観察ては,ウイルスの増殖に先立ちpermuclear gap,あるいは,r-ERの一部が著明に発達し,嚢状となり,感染時間の経過とともにこれらの内腔には単層の小胞状,小管状構造が出現し,外被を持った直径45～50mμの成執ウイルス粒子が観察された.感染末期には,嚢状構造は崩壊した原形質より風船様に遊離し,その構造の一部には外側に直径23～28mμの電子密度の高い粒子と,その内腔には,膜に接した成執ウイルス粒子を観察した.
6.免疫電顕法による観察ては,感染後48時間には,成熟ウイルス粒子は勿論,細胞膜表層,および小管状,小胞状構造の内面にも抗原陽性部位が認められた.

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Japanese encephalitis (JE) is a nonpurulent encephalitis caused by the Japanese encephalitis virus(JEV)which belongs to the group of Flavivirus of the Togaviridae. JEV, the etiologic agent of this disease, is widely distributed over a large area of Asia, from Siberia to India. Within the area there are two geographically and meteorologically separable regions, each of which has a distinct pattern of virus dissemination. In countries of tropical climates, JEV is presumed to be endemically maintained throughout the year and human infections occur sporadically. In countries of temperate climate including Japan, JE outbreak occurs about the middle of August every year. The scale of the epidemic varies from year to year. Two epidemics of large scale have been recorded in the 1930s and 1940s.
Although data and knowledge have so far been accumulated on the transmission route of JEV for human infection, the ecology of JEV and the mechanism of epidemic outbreaks of JE patients still remains to be solved. Therefore it is not possible, at this time, to predict the future dissemination of the disease. In order to obtain knowledge useful for controlling JE epidemics, epidemiological and ecological studies of JEV transmission were carried out in Shiga Prefecture between 1965 and 1973, and the following results were obtained.
The rate of human incidence was the highest in 1966 and decreased with a few fluctuations. No patient was recorded in 1971. Together with the foregoing data (1955-1964), a typical pattern of human JE epidemic was revealed.1) The main epidemic follows a presubepidemic and is followed by a post-subepidemic.2) The scale of each epidemic is repeated by eight year interval.
Swine infection correlated with the human incidence. In the year when swine infection occurred earlier and antibody titer of swine sera were higher, more human patients were recorded and vice versa.
The number of adult femaleCulex tritaeniorhynchustrapped also correlated with both human incidence and swine infection. That is, the year when many mosquitoes were trapped was a year when many human patients were recorded.

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In the preceding paper, the author reported that the decrease in number of femaleCulex tritaeniorhynchus trapped paralleled the decrease in human incidence. In an attempt to elucidate the factors affecting the decrease of mosquito, meteorological conditions and annual consumption of weeding agents and insecticides were analyzed. The following facts were noted.
In the years (1966,1967) when many mosquitoes were trapped, the mean temperature of the first decade of March (4.6-7.9°C) was higher than that of ordinary year (4.0°C). In 1969-1973, it was lower than the ordinary year and the number of mosquitoes was fewer than before.
Throughout the period, consumption of two insecticides, γ-BHC and Sumithion, was increased and negatively correlated with the mosquito population, while the consumption of other insecticides or weeding agents was the same or decreased. All these findings and the literature suggest that the mean temperature of the first decade of March and the consumption of two insecticides are the factors which influence the mosquito number.

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In the ecology of Japanese encephalitis virus (JEV), the host animal for the virus during the non-epidemic season still remains uncertain.
In this paper, the author sought to determine the animal species which play an important role in the transmission of JEV just before an epidemic.
Sera were collected from wild herons, rats, snakes, lizards, and domestic fowls. Antibody-free dogs and rabbits, placed at two points as sentinels, were bled biweekly for two years. A prevalence of hemagglutination inhibition (HI) antibody was shown in dogs, rabbits, domestic fowls, herons and snakes, while not in rats and lizards. Seasonal transitions of HI titer chased in the sera of sentinel animals. The serum of each sentinel animal showed ascending HI titer late in July to early in August. All these results and other literature suggest that hibernating animals may play an important role as viral host during the winter season. Migratory birds may transmit JEV from endemic areas, such as South East Asia, to Japan and these two groups of animals may transmit JEV and cause epidemics in Japan.

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This final paper summarizes the results of epidemiological studies of Japanese encephalitis in Shiga Prefecture between 1965 and 1973, described in the preceding three papers. Based on the results obtained in these study series, certain possibilities regarding epidemic outbreaks of Japanese encephalitis in the future was suggested.
Throughout the period of the investigation, a gradual decrease in human incidence was observed. This correlated well with the decrease of vector mosquitoes (Culex tritaeniorhynchus). Changes in meteorological conditions and the amounts of two insecticides sprayed were discussed as explanatory reason for the decrease of vector mosquitoes.
In spite of the parallel decline of human infection and mosquito number, JEV infection rate in swine born in a nonepidemic season reached 100% the next summer season, sooner or later. There must be certain animals which originally supply JEV to swine. Some of them are hibernating animals in which Japanese encephalitis virus (JEV) passes the winter, and others are migratory birds by which JEV is transported from endemic areas to Japan.
Consequently, it can be said that the possibility of epidemic outbreak of Japanese encephalitis still remains in Japan. Care must be therefore taken to exclude the epidemic factors by extermination of vector mosquitoes and maintaining social immunity by vaccination of both man and domestic animals.

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Effects of maltose, glucose and vitamin B₁ on the central nervous system and intestinal movement were electrophysiologically examined by use of rabbits.
1. On the arousal reaction and the evoked muscular discharge induced by stimulation of the brain stem reticular formation, and on the inhibitory effect of arousal reaction toward intestinal movement, activated vitamin B₁,50%glucose and maltose mainly showed inhibitory effect. On the contrary, they showed facilitatory effect on the intestinal movement.
2. On the evoked muscular discharge of fore and hind limbs induced by stimulation of the cerebral cortex, the small amount of 50%glucose and maltose showed inhibitory effect, on the contrary, the great amount of 50%glucose and maltose showed facilitatory effect, and activated vitamin B₁ showed both inhibitory and facilitatory effect in which no certain tendency was recognized.
3. On the evoked muscular discharge induced by stimulation of the hippocampus,50% glucose and maltose showed facilitatory effect, on the other hand, activated vitamin B₁ mainly showed inhibitory effect. Thus carbohydrate solution and activated vitamin B₁ showed opposite effects.
4. On the early components (N₁, N₂) of afferent average evoked potentials of the cerebral cortex induced by the sciatic nerve stimulation, activated vitamin B₁,50% glucose and maltose showed facilitatory effect. On the contrary, they showed no significant effect on the late components (N₄, N₅).
5. On the nociceptive reflex induced from the tibialis anterior muscle due to stimulation of the tibial nerve, activated vitamin B₁,50%glucose and maltose showed facilitatory effect.
6. On the H wave induced from M. gastrocnemius tibialis due to stimulation of the sciatic nerve, maltose,50%glucose and activated vitamin B₁ showed facilitatory effect in which activated vitamin B₁ and 50%glucose showed stronger effect than maltose. On the M wave the activated vitamin B₁ and 50%glucose showed facilitatory effect, when compared with the effect on the H wave, effect on the H wave is considered stronger. Maltose showed no significant effect on the M wave.
7. Activated vitamin B₁,50%glucose and maltose increased more than 30% the threshold of arousal reaction and evoked muscular discharge due to stimulation of the reticular formation.5 %glucose slightly increased the threshold.
From these completed results, it is revealed that activated vitamin B₁ and carbohydrate (administrated at the same time) on one hand, showed facilitatory effect on the afferent pathway and the intestinal movement, but that on the other hand, they might show the inhibitory effect on the arousal reaction. The countermeasure has been discussed.

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Both sexes of Sprague-Dawley JCL rats, aged 8∼9 weeks and weighing 200∼250g, were injected subcutaneously by either 100mg/kg or 200mg/kg of Sulpiride daily for 1 month. In females treated with Sulpiride, a marked proliferation with severe milk secretion could be observed on the mammary glands. The weight of abdominal mammary fatty pads increased twice. Their endocrine glands could reveal no prominent changes. The sexrelated organs, especially the uterus, were somewhat atrophic. In males injected by Sulpiride, the similar changes could be detected, but they were weaker than females.
Adult female Sprague-Dawley JCL rats, some of which were given orally 1000mg/kg of Sulpiride daily in definite periods, were checked their vaginal smears every early morning. All treated rats revealed to begin a continuous diestrous state in a week after the first intubation and to stop a diestrous condition in a week after the last intubation. The mammary glands of the females treated for 70∼90 days were prominently proliferative and hypersecretory, and they revealed a severe milk stasis with yellow pigment deposition. A marked atrophy with fibrosis could be detected in the mammary fatty tissues 3 weeks after the ceasation of daily intubation, when the cycles of vaginal smear returned to be regular.
By means of Khazan et al. 's index, the mammotrophic activity of Sulpiride was compared with that of Chlorpromazine. The effect of subctaneous injection of Sulpiride,100mg/kg, daily for 5 days upon the estrogenized females was corresponded to that of Chlorpromazine,5mg/kg.
Electron-microscopic observations were carried out on the mammary glands of diestrous female rats within 1 day after a subcutaneous injection of Sulpiride,100mg/kg. Secretory pictures, e. g. increase of ribosomes, many secretory granules in the swollen ER, Golgi apparatus and lumen, could be detected 6 hours after the injection. These findings were more prominent 12 and 24 hours after the single injection. The early effect -Irug were recognized morphologically in a week of the daily injection by proliferation of the epithelium with intercellular gaps, and by desquamation of superficial acinar cells. The content of ribosomes in the cytoplasm increased. Many bigger fat droplets and large swollen mitochondria exhibited in the perikaryon. Retention of secretion followed on the continuous stimulation. Such a milk stasis evoked definite degenerative and atrophic alterations in some mammary parenchyms.

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No mammotrophic action of Sulpiride could be detected on both the hypophysectomized rats injected subcutaneously by either 100mg/kg or 200mg/kg of the drug daily for 2 weeks and the ovariectomized rats injected subcutaneously with 100mg/kg of the drug daily for 2 weeks. Daily injection of Estradio1-17β,10γ, into the hypophysectomized ones could not induce to reveal the action of Sulpiride, but it into the ovariectomized animals could reveal the mammotrophic action of the drug. Bilateral adrenalectomy could not disturbe to reveal the action of Sulpiride.
Radioimmunoassay for prolactin was performed on the serum and pituitary tissue of the diestrous females, which had been checked regular sex cycles in 2 weeks,6,12 and 24 hours after the subcutaneous injection of Sulpiride,100mg/kg. A single dose of the drug elevated the serum prolactin levels to about 3 times that of the controls 6-12 hours after the injection, when the pituitary contents were somewhat lower than that of the controls. The level in serum still elevated about 2.5 times 24 hours after the administration, when the content in the adenohypophysis increased 2 times that of the controls. On the sera of the rats injected daily, prolactin levels went down gradually and returned to normal level more than 7 days after the first injection.
Distribution of the radioactivity were determined on the mammary fatty pad, ovary, adrenal gland, hypophysis, hypothalamus and brain of the rats 1/2,1,3 and 24 hours after the intramuscular injection of ¹⁴C-Sulpiride,2mg/kg, respectively. Higher activity in the pituitary tisssues might suggest the direct action of the drug into the adenohypophysis.
In the rabbits, Chlorpromazine induced lower body temperature but Sulpiride,100mg/kg, could not induce lower body temperature in 6 hours after the subcutaneous injection.
In summary, Sulpiride would act the adenohypophysis directly, and induce hypersecretion of prolactin in short time after the administration. Stimulation of prolactin could develope a proliferation with secretion on the mammary glands of the estrogenized animals. On the process of mammotrophic action with Sulpiride, the brain, adrenals and probably hypothalamus might have no direct actions.

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In order to investigate a mechanism of perinatal anoxic brain damage the cerebral cortex of newborn rat with anoxic episodes in intrauterine life was examined mainly by using electron microscope.
After anaesthesia induced by intramuscular injection of sodium nembutal, pregnant rats of 18th gestational day were operated on the abdominal wall to expose a small portion of the uterus. Unilateral uterine arteries were clamped by forcets for 2 minutes, and then 1 minute-relaps in clump-free condition was followed. The same procedure was repeated 5 times by means of Nakamura's method.
After the uterus was restored into the abdominal cavity, the abdominal walls were sutured up. Thereafter, the gastation was resumed for the successive 3 days.
Newborn rat brain was cut out and immediately fixed in 2 %glutaraldehyde after birth. The central gyri of the brain tissue was examined by electron microscopy on one day,7 days,2 weeks,3 weeks,4 weeks,5 weeks,6 weeks,7 weeks and 8 weeks after birth, respectively and the following results were obtained.

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In the report,4 patients associated with double aortic arch are presented. Two patients underwent surgery are 28-year-old woman and 1-year and 8-month-old girl. In the former, the left sided aortic arch is atretic with left ligamentum arteriosum (Edwards subgroup IB₁), and the atretic aortic arch and the ligamentum arteriosum are divided. In the latter, the left sided aortic arch is also atretic (Edwards subgroup IB₂), which is divided. Moreover, in this case, an associated cardiac malformation of partial anomalous left upper pulmonary venous drainage into innominate vein with pulmonary stenosis is demonstrated. Postoperative courses of both cases are uneventful. The other 2 patients without surgery are 64-year-old man (Edwards subgroup IA) and 50-year-old man (Edwards subgroup TB or III B). As far as we can ascertain from viewing the literature, the older patients over 50 years old are 18 inclusive of the presented 2 patients.

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