Abstracts of Papers Presented

at the

Twentysecond Annual Meeting

of the

Japanese Teratology Society

Tokyo, Japan July 8-9,1982

CONTENTS
Symposium on Congenital Limb and Vertebral Anomalies ................................ Symposium on Maternal Environment and Congenital Anomalies ........................... Lectures by Invitation
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Papers Presented from Platform and Read by Title ...................................

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Symposium on
CONGENITAL. LIMB AND VERTEBRAL ANOMALIES

YASUDA, M., Department of Anatomy, Hiroshima University School of Medicine, Hiroshima. Recent topics on morphogenesis of the limb and the vertebral column. Hypotheses have been developed recently to explain the morphogenesis of periodic patterns such as finger rays and somites. For example, the positional information theory by Wolpert, and the clock and wavefront model by Cooke and Zeman have attracted attention of many developmental biologists, and have been tested by various experiments. Cell movement and aggregation are important phenomena in limb and vertebral morphogenesis. Recent studies have revealed that extracellular matrices and cell surface properties are critical in controlling these phenomena. Findings have shown the relationship between cyclic A M P and the distribution of fibronection and hyaluronate in the process of mesenchymal condensation. The 3rd International Conference on Limb Morphogenesis and Regeneration was held in Storrs, Connecticut, from June 27 to July 2, 1982. Many current topics were presented at the Conference. This presentation deals with some of the topics.

cal, pharmacological and developmental biological methods have enabled u s to explore the mechanism of teratogenesis in detail. However, the mechanisms and pathogenesis of most human malformations are still unknown. It is expected that in the future, explorations and advances are to be made, chiefly by molecular teratology, in understanding the etiology and mechanisms of human malformations including those which are now explained as multifactorial in origin.

SHIOTA, K., Congenital Anomaly Research Center, Faculty o f Medicine, Kyoto University, Kyoto. Detection of etiologic factors: Procedures and problems. For the detection of etiologic factors in human malformations, epidemiologic sutdies and surveillance in human population as well as analytical studies of individual patients are essential. However, surveillance o r monitoring systems have their ljmits and problems. Since most human teratogens have been first speculated by practicing physicians, they are encouraged to give case reports on suggestive findings which should lead to further investigation in detail. The promotion of dysmorphology is also needed. An increasing number of potentially teratogenic agents have been identified using in vivo and in vitro animal studies. Employment of new research techniques such as whole embryo cultures and of biochemi-

MIURA, T., Department of Orthopaedic Surgery, Branch Hospital, Faculty of Medicine, Nagoya University, Nagoya, Aichi. A clinical approach for congenital hand anomalies. The clinical and roentgenologic features of split hands, syndactyly, radial and ulnar ray deficiencies, and brachydactly were studied. Clinical findings make clear that the abnormal development of AER, abnormal extension and delayed involution of AER, are considered to be responsible for the formation of some rongenital hand anomalies after the formation of the hand plate. If there is an abnormal distrihution and arrangement of mesoderm due to the abnormality of AER, the hand plate will develop abnormally, resulting polydactyly, syndactyly or a split hand (oligodactyly). In these anomalies induced by abnormalities of AER, abnormal findings in limbs are limited to the portion distal to the wrist. The critical period for a transverse deficiency may be earlier than the period of webbing or union of the digital rays. The critical period of a radial ray deficiency may be extended throughout the period of development of limb buds. The critiral period of webbing or union of the digital rays may be earlier than the period of mesenchymal condensation; and in the same period, a split hand can be formed as a result of the union of the digital rays. If necrosis of the mesenchymal tissue was the dominant feature in a split hand and syndactyly, these hand anomalies would be accompanied by hypoplastic abnormalities in the proximal portion, as are symbrachydactyly or transverse deficiencies.

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ONOMLIU, T., Department of Orthopedic Surgery, Osaka Medical College, Takatsuki. Osaka. Congenital anomalies of the spine: Problems between experimental animal models and clinical cases. Spinal malfonnations, experimentally induced in rats, were observed and compared with congenital spinal anomalies of humans which are frequently seen in orthopedics. Model animals suggest a reasonable classification of the varieties of congenital spine deformities, associated anomalies hidden in or out of the skeletal system, chronological changes of the malformed spine in the postnatal growing age and suitable therapeutic management of individual cases. Clinical material is limited mainly t o postural deformities due to congenital vertebral anomalies, including scoliosis and kyphosis. The variations observed in the incidence of spinal anomalies in drug-treated rats, and the high mortality rates of the malformed offspring make clarification of the causative mechanism difficult.

YAMANE, T., Department of Orthopedic Surgery, School of Medicine Teikyo University, Tokyo. Management of spina bifida. Ninety-nine patients with spina bifida were reviewed. There were 54 myelomeningocele patients, 26 meningocele patients and 19 spina bifida occulta patients with lipomas or dimples who visited o u r spina bifida clinic from 1967 to 1982. Immediate closure operations were performed for open myelomeningocele conditions and release operations were done occasionally for other types of spina bifida according to urinary dysfunction, feet deformities and the findings of metrizamide myelography. Lesions of the lumbo-sacral and sacral levels were seen in about 86% of the cases. The most frequent orthopedic problem was foot deformities such as clubfeet and calcaneus feet. Although most of the 115 deformities could controlled by splintages or braces, 23 foot deformities were treated with surgery. Paralytic dislocation of the hip was also an important problem. Two patients were surgically treated for a sitting disturbance. Spinal deformities such as scoliosis or kyphosis tended to be exaggerated in children when 4 or 5 years old. Two scoliotic patients had difficulty in stabilizing an erect position of the trunk while in a wheel chair. In our clinic, spina bifida patients are checked for urinary function, neurological levels and deformities of the trunk and extremities at monthly intervals and treated when necessary.

anomalies with malformations of the gastro-intestinal tract. Seven hundren and forty-three newborns with gastro-intestinal anomalies were analysed. The incidence of associated vertebral anomalies was high in the anorectal anomaly group ( 2 4 . 2 % ) , esophageal atresia group (12.9%) and abdominal wall anomaly group (11.9%). The associated limb anomalies were also frequently observed in these three groups. Hirschsprung's disease and hypertrophic pyloric stenosis showed a low incidence of associated vertebral and limb anomalies, contrary to the fact that the two are known as hereditary diseases. We have encountered three cases of VATER association (Vertebra, Anal atresia, Tracheoesophageal fistula. Esophageal atresia and Radius or Renal malformation). However we have not seen notochord syndrome (intestinal duplication with vertebral malformation), in which association is easily explained from the embryological basis. Special attention should be paid on the sacral malformations associated with anorectal anomalies. A sacral malformation is frequently accompanied by a sacral nerve defect, which results malfunction of defecation after radical operation for the high type of imperforate anus. Hence the sacral bone should be investigated and evaluated before the radical pull through operation.

. OHAMA. and T. IZAWA, TSUNODA, A., Y Departments of Surgery and Orthopedics, Kanagawa Children's Medical Center, Yokohama, Kanagawa. Association of vertebral and limb

TSUCHIYA, K., Kanagawa Rehabilitation Center, Nanasawa. Atsugi, Kanagawa. Thalidomide embryopathy and its allied conditions in Japan. The number of thalidomide patients born in Japan and surviving to adulthood was 309, of which two died for unrelated reasons. Therefore, the number of patients at present is 307. Clinical features of thalidomide embryopathy ) the are anomalies o f : 1) the extremities, 2 ears and eyes and 3) the internal organs. With one exception, all patients suffered from anomalies of the upper extremities. The one exception had lower extremity involvement. Limb anomalies consisted of 1) amelia, 2) phocomelia, 3) severe ectromelia 4 ) ectromelia, 5 ) hypoplasia or defect of the thumb, and 6) thenar muscle hypoplasia. Non-thalidomide induced malformations, distinguishable from thalidomide malformations, are: 1) Holt-Oram syndrome, 2) Fanconi syndrome and 3) congenital thrombocytopenia with radius hypoplasia. These three anomalies are genetically transmitted. 4 ) Congenital radius hypo- or a-plasia is non-inheritable. In Japan, thalidomide malformations ceased in 1964. but was followed by non-thalidomide malformations, whose causes were unknown. In Yokohama, fine cases of clubhand with radius hypoplasia plus thumb hypoplasia or defect were observed by the author. A survey by questionnaire revealed 40 similar cases in Japan. The main difference between non-thalidomide and thalidomide induced malformations was that the drug had a marked tendency to cause bilateral abnormalities.

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Symposium on

MATERNAL ENVIRONMENT AND CONGENITAL ANOMALIES

OMORI, Y., Diabetes Center, Tokyo Women's Medical College, Tokyo. Congenital malformations in the infants of diabetic mothers (IDM). Mayer named "embryopathia diabetica" as the abnormalities of fetuses of diabetic mothers. These abnormalities, including congenital malformations (CM), are seen when diabetics are unable to adequately control their insulin levels. CM and respiratory distress syndrome are still the main causes of death among IDM. I analysed the frequency, type of malformations, and relationship of maternal diabetes treatment with regrads to 110 babies and 89 cases of pregnant diabetics. CM in the IDM in Japan over a ten year period was also surveyed. To elucidate the mechanism of CM, the influence of maternal hyperglycemia and hypoglycemia on the fetuses in diabetic animals was observed. The frequency of CM was 16.4% (18 oLt of 110 newborns) in our hospital compared with 1 . 5 % for controls, Although most of the IDM had minor abnormalities, three newborns died due to CM. 78% of mothers who had infants with CM showed poor insulin control in the first trimester. In a Japan survey, CM was 6.2% 4 4 out of 1042 IDM. The types of CM seen in IDM are distributed throughout all organ systems. These results and studies with animal models suggest that CM is not related to maternal hypoglycemia but hyperglycemia.

manifestation of non-stochastic somatic effects which have been recognized are: 5 rad for resorption of preimplantation embryos; 5-10 rad for acute cytological changes such as pyknosis, cytoplasmic degeneration and mitotic delay; 5 rad for increasing frequency of spontaneous minor anomalies of the skeleton; 15-20 rad for malformations of the eye, brain and spinal cord; 20-25 rad for histogenetic and functional disorders of the central nervous system; and 20-25 rad for impaired fertility. Pregnant women who are subject to X-ray examinations are concerned about the potential hazards of radiation to their offspring in utero. The above experimental findings suggest that the possibility of non-stochastic somatic effects of diagnostic radiation on human embryos and fetuses is extremely low, and probably negligible, given the proper dose control measures. Possible effects, which should be considered for risk evaluation of diagnostic exposure, are carcinogenic and mutagenic induction. (Supported by Scientific Research Grants No. 258070 and 238017 from the Ministry of Education, Science and Culture, Japan)

KAMEYAMA, Y., Research Institute of Environmental Medicine, Nagoya University, Nagoya, Aichi. Low-dose X-radiation and congenital anomalies. Among radiation effects on developing embryos and fetuses, the occurrence of germinal mutations due to exposure of the gonads and postnatal manifestations of neoplasms are considered to be stochastic effects from the aspect of radiation protection. Somatic effects such as teratogenic and embryo-toxic effects, however, can be regarded as nonstochastic with threshold doses. In experimental teratological studies with mice and rats, the lowest radiation doses for

TANIMURA, T. Department of Anatomy, Kinki University School of Medicine, Sayama-cho, Osaka. Congenital anomalies induced by pharmaceutical drugs. It is estimated that less than 2% of the congenital anomalies can be attributed to chemicals including drugs. However, developmental toxicity of drugs have the following characteristics. 1. A considerable number of new compounds are introduced into the market every year after reproduction tests in animals. 2. Drugs are intentionally administered, often subchronically at a relatively high dose. They are given after consideration that potential benefits to a pregnant mother outweigh possible hazards to both the mother and her unborn child. 3. Congenital anomalies induced by drugs can be prevented if they are withdrawn from the market. Therefore, a stronger administrative judgement is required in evaluating the hazards of drugs to human

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development as compared to not only environmental chemicals but also physical and biological developmental toxicity agents. In this symposium, first, methods for detection of developmental toxicity of drugs are presented. Basic principles, significance of animal studies and human surveys (case reports and epidemiological studies) are discussed. Secondly, critical reviews are presented for some drugs which have recently become the subjects of controversial discussion. These include Bendectin, diethylstilbestrol, progestogens, vaginal spermicides and anticonvulsants. Finally, various topics on the balance between benefits and risk are discussed.

an experimental rat model was employed. A decrease in cerebral weight and DNA synthesis was observed when dams were ingested throughout pregnancy, but with no reduction of fetal body weight, litter size, or maternal weight. The fetal surviving rate was related to maternal serum caffeine levels. In conclusion, it would he prudent for women planning to have a baby to limit their alcohol and caffeine intake.

TANAKA, H., Division of Child Neurology, National Center for Nervous, Mental and Muscular Disorders, Kodaira, Tokyo. Congenital anomalies induced by maternal alcohol and caffeine. Of the preventable causes of congenital abnormalities, maternal drinking of alcohol and caffeine during pregnancy was investigated. Japanese cases tended to show a mild effect of maternal ethanol comsumption, especially on growth retardation, suggesting some timing discrepancies between growth and mental retardation. To study the cause of the fetal alcohol syndrome (FAS), an experimental rat model was employed. An increase in perinatal mortality, a decrease in fetal body and cerebral weights and decreases in DNA, RNA and protein contents or syntheses in cerebrum were observed. The specific activity of 2 ' , 3'-cyclic nucleotide 3'-phosphohydrolase in developing rats showed accelerated growth, hut RNA content remained at low levels. Hypoplasia, as the characteristic feature of FAS, was related to maternal serum and fetal body ethanol levels. It occurred prior to the development of ethanol-induced hypoglycemia in late pregnancy, so hypoglycemia was not the main cause of FAS. Low levels of maternal serum insulin and cerebral carbonic anhydrase and zinc were observed. Zinc administration to rats on alcohol diet resulted in better effects on fetal body weight and preventing resorption than the administration o f ethanol alone but no better than control. To determine whether maternal caffeine ingestion was a risk factor in brain development,

KITAGAWA, T., Department of Pediatrics, Nihon University School of Medicine, Tokyo. abnormalities from maternal metabolic disorders and endocrine diseases. Results of a survey and our study on fetal and neonatal abnormalities associated with maternal metabolic disorders and endocrine diseases are reported. DQs in two histidinemic cases of maternal histidinemia were within the normal range. It has been reported that three cases of maternal histidinemia with normal histidine metabolism and ten histidinemic patients of maternal histidinemia including the above two cases showed normal DQs. DQs of four phenylketonuria (PKII) patients of maternal PKU and five cases of maternal PKU with normal phenylalanine metabolism have been reported in Japan. Two of the maternal PKUs had normal phenylalanine metabolism and DQs. One patient died from multiple congenital anomalies accompanied by hypoplastic left heart syndrome while the mother was on a low phenylalanine diet beginning at six weeks of gestation. It has been shown that the most common maternal endocrine diseases were maternal thyroid disorders. Nine to fourteen per cent of maternal thyroid disorders showed neonatal abnormalities. Uncontrolled Cushing syndrome, pheochromocytoma, pituitary gigantism and acromegaly in the mother were responsible for occasional spontaneous abortions, premature labor, fetal death and neonatal anomalies. The high frequency of neonatal abnormalities were associated with uncontrolled hypoparathyroidism in the mother. Therefore, early treatment of metabolic disorders and endocrine diseases in pregnant women is important for the prevention of severe fetal and neonatal abnormalities.

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Lectures by Invitation

OHAMA, K., Department of Obstetrics and Gynecology, Hiroshima University School of Medicine, Hiroshima. Prenatal selection of conceptuses with chromosome abnormalities. Recent cytogenetic studies on spontaneous and induced abortions indicate that almost half of the fertilized ova are chromosomally abnormal and that 90% of the abnormals are eliminated before the pregnancy is recognized. Around 5% of conceptuses at the beginning of clinically recognizable pregnancy are chromosomally abnormal. Of these 5 % , half are autosoma1 trisomies involving chromosomes 2 through 22. Trisomy 1 has never been encountered. Among the trisomies, trisomy 16 is most frequent, accounting for 30% of all trisomies. Trisomies 2 1 and 22 are the next most common, 8 are being 10% each, and trisomies 13 and 1 5 % each. Monosomy X is found in 19% of the abnormals, triploidy is 17% and tetraploidy 5%. More than 90% of such abnormal conceptuses later abort spontaneouslv, most of them during the first trimester of pregnancy. Thus, only 0.6% of liveborns are chromosomally abnormal. Prenatal selection rates of these chromosome abnormalities are: trisomy 21 : 77%, trisomy 18 : 94%, trisomy 13 : 96%, monosomy X : 99% and triploidy : 99.8%. Most of the abortuses with lethal chromosome abnormalities are either empty sacs or totally disorganized embryos. They stop developing at 6 weeks of embryonic age or earlier. On the other hand, apparently normal fetuses are occasionally found among conceptuses with trisomy 21, monosomy X and triploidy. In conclusion, spontaneous abortions are an efficient device for the elimination of abnormal conceptuses.

one dup (dir or inv) and 6 psu dic were detected in the author's laboratory, although only one case each of inv dup, dup and psu dic was found during the preceding 7 years. The incidence of both inv dup and psu dic may be much higher than what is generally believed. The Cd-staining technique reveals only an active centromere (Nakagome, 1976). The centromeres of aged individuals were examined by this technique to evaluate their possible inactivation as the cause o f non-disjunction.

INOLNE, E., Institute for Developmental Research, Aichi Prefectural Colony, Kasugai, Aichi. Genetic basis of normal human behavior. The notion that human-specific behavioral patterns are genetically determined is still an untestable hypothesis. Polymorphic Mendelian behavioral traits are limited to those of sensation. Association between sex chromosome anomalies and quasi-normal behavioral patterns is never complete. Most quantitative genetic analyses have merely provided an array of heritability figures. Our observations and experiments on twins have revealed a genetic basis for spontaneous, undifferentiated, unconscious and autonomous bodily and facial movements and sense of time span. Non-dominant cerebral hemispheres in monozygotic (MZ) twins indicated a higher correlation than the dominant half in the amount of beta waves reflecting arousal activity of the brain suggestin$ the presence of genetically determined neural patterns. The above findings were recently confirmed by reports on dichotic listening and sensing of the center of gravity while in a standing posture in MZ twins.

NAKAGOME, Y., Department of Human Genetics, National Institute of Genetics, Mishima, Shizuoka. New chromosomal syndromes and functioning kinetochore in aged individuals and its effects on non disjunction. About 190 different types of autosomal abnormalities have been described. By the use of newer techniques, e.g., distamycin-DAPI, Ag-I and high-resolution banding, an increasingly large number of cases with unusual structural rearrangements are being identified. In the past 3 years, 6 cases of inv dup, one dir dup,

GOMIBUCHI, M., Japan Association of Maternal Welfare, Tokyo. Surveillance systems for congenital malformations in Japan and their problems. The Japan Association for Maternal Welfare has been conducting surveys of congenitally

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malformed newborns reported in 130 hospitals in Japan since January, 1972. Many similar investigations have been performed and reported in Japan. For effective surveillance, it is necessary to continue the investigation for a long period under the same conditions. Our surveys have been performed more than ten-years with more than 1,000,000 babies. Statistical analyses of several congenital malformations frequent in Japan have been performed. Problems in such surveys are reviewed.

normal shape using available tissue properly and effectively. For the future of the handicapped children, it is the duty of the doctor to give moral support to the family. 2 ) When grown up, the patient can be operated once more f o r further improvement, by means of fine revisional techniques for the residual blemishes. It may be necessary to transplant the tissue from another part of the body, o r to insert a prosthesis.

NAKAJIMA, A., Department of Ophthalmology, School of Medicine, Juntendo University, Tokyo. Epidemiology of congenital eye anomalies in Japan. __ The majority of congenital eye anomalies affecting vision is easily recognized. Retinopathy has often been a problem recognized in premature babies. According to Harada, retinopathy of premature infants was found in more than one third of the blind children horn in Tokyo between 1965 to 1973. Since 1973 there has been an abrupt decrease in the incidence of retinopathy because of improved care of premature babies. Every five years since 1954, surveys on probable causes of blindness in children in schools for the blind have been conducted. The proportion of retinopathies of premature infants was 1 % in 1970, 5% in 1975 and 10% in 1980. There was a steady decrease in corneal diseases, and a relative increase of microphthalmos, cataract, retinal degenerations and optic nerve atrophies. The analysis for etiology using family relation and consanguinity has suggested that genetic factors are important. However, environmental factors are also important under some conditions. The importance of early detection and treatment was stressed in buphthalmos, retinoblastoma and other conditions.

HIGURASHI, M. and K. IIJIMA, Department of Health Sciences, Medical University of Yamanashi, Yamanashi, and Department of Maternal and Child Health, University of Tokyo, Tokyo. Newborn infants and chromosomal aberrations. During the past decade several chromosome surveys on newborn infants in North America and Europe have been reported. Recently two o r three surveys including ours were reported in Japan. The incidence of chromosomal aberrations in newborn infants based on these reports are presented. The screening methods for chromosomal aberrations for newborn infants, the limitations of these screening methods, and the transition of some clinical manifestations on newborn infants with chromosomal aberrations are discussed.

TANGE, I., Plastic Surgery Unit, School of Medicine, Juntendo University, Tokyo. Facial anomalies and their improvements by plastic surgery. A total of more than 3,000 facial anomalies have been operated on in the last twenty years in this Unit. Most of the cases are ordinary cleft lip with or without cleft palate. But the rare facial clefts, such as median, oblique, and transverse cleft and their variations which were recently classified by Paul Tessier (1973), are also included in this series. The operations were performed based on aesthetic points. The following was concluded: 1) At the time of primary repair performed during infancy or early childhood, it was essential to avoid unnecessary loss of the tissue. It was important to build up the

KIMURA, M., The Institute of Public Health, Health and Welfare Ministry, Tokyo. Birth defect monitoring systems and epidemiological studies. Monitoring systems or services primarily aimed at checking an unusual occurrence are widely applied in society. The movement of human vital events, however, shifts slowly and the level of unusual incidence seems to be rather low. National statistical services on birth defects started in the 1950's and eventually developed into monitoring services including epidemiological intelligence in the 1960's. In 1974, scientists of 13 countries created a clearinghouse service of information of their monitoring systems. The 31st WHO General Assembly in 1978 reported that it recognized that birth defects contributed t o early mortality and detracted from the quality of life in all societies. There is a need for further information, in particular their relation to environmental hazards. The European Community recently started its own project "EUROCAT" with 18 designated institutions to monitor the outbreak of birth defects within Europe. From the 1982 yearbook of ICBMS, several topics including femur reduction deformity in the Alps-Rhone, types of Down syndrome, Ebstein heart anomalies and classification of multiple malformations were presented.

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SUZUKI, S., Department of Obstetrics and Gynecology, School of Medicine, Keio University, Tokyo. Recent advances in the mechanism of fertilization. Reproductive medicine has progressed and many clinically important problems have been resolved. However, the mechanism of fertilization, which is the most basic and fundamental

phenomenon in human reproduction. has not yet been clarified. We have investigated the morphological and functional characteristics of the human oocyte in vivo and in vitro. Observations of the human oocyte at o r before fertilization in vitro including ultrastructual changes will be discussed.

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Papers Presented From Platform and Read by Title

NAKAO, Y., R. UEKI, T. TADA, I. IRITANI and H. KISHIMOTO, Department of Pathology, Nagoya City University Medical School, Nagoya. Role of the lung buds on development of the dia-

phragm.
Previously, we reported that 2,4-dichlorophenyl-pnitrophenyl ether (TOK) induced congenial diaphragmatic hernia (CDH) in mice. The same teratogenic effects have now been observed in rats. Diaphragmatic defects and hernias were predominant at the posterolateral portion of the right side of the diaphragm in contrast with the findings in mice where the left side (posterolateral portion) was affected most frequently. We also studied development of the diaphragm, lung and some mesodermal-mesenchymal tissue using serial sectioning of whole bodies of embryos and fetuses. The following conclusions are presented: 1. TOK affects development of mesodermalmesenchymal tissue of embryos and fetuses in

day 13, 15, 17, 19 OK 21 of gestation and serial sections were examined using light and electron microscopy. Light microscopic examination revealed maldescent and hypoplasia of the thymus i.e., reduced in size, lobulation and a number of small lymphocytes in the cortex. Electron microscopic studies showed that the cortex of the thymus contained large-, medium- and small-sized cells. The large cells had an immature structure and were located adjacent to the basement lamina. The small cells were lymphocytes and were aggregated near the medullary region. It is concluded that the hypoplastic thymus is the result of delay in the differentiation of thymic lymphocytes.

utero.
2. Normal development of the lung buds at an early developmental stage is essential to normal development of the primitive diaphragm. 3 . Lung hypoplasia associated with CDH in neonates is not secondary to the herniation of abdominal viscera into the thoracic cavity. Insufficient lung bud development occurs primarily because of abnormal mesodermalmesenchymal tissue development and differentiation at an early developmental stage. Therefore, growth retardation of the lung persists after completion of the primitive diaphragm.

IWASAKI, K., K. KAWAMOTO, T. MATSUO and T. IKEDA, First Department of Pathology, Nagasaki University School of Medicine, Nagasaki. Bis-diamine induced hypoplasia of the thymus. Bis(dichloroacetyl)diamine, fertilysin, has been shown to produce defects of the branchial arch and pharyngeal pouch by oral administration into pregnant rats. For the purpose of investigating abnormalities of the thymus, bis(dichloroacety1)diamine was given to pregnant Wistar rats by gastric tube on day 10, 11 OK 12 of gestation. Fetuses were removed on

LEE, J. Y., N. OKAMOTO, Y. SATOW and H. SLIMIDA, Department of Geneticopathology, Research Institute for Nuclear Medicine and Biology, Hiroshima University, Hiroshima. Persistent truncus arteriosus, hypoplasia of the pulmonary trunk and aortic arch anomalies induced by Bisdiamine in rats. The authors previously induction of . reported . persistent truncus arteriosus (PTA), hypoplasia of the pulmonary trunk (HPT) and aortic arch anomalies by Bisdiamine given to rats on the 10th day of pregnancy, and the relationship between dosage and frequency of anomalies (21st Annual Meeting of the Japanese Teratology Society, 1981). In the present study, fetal hearts with PTA, HPT (or atresia) and ventricular septa1 defects (VSD) were examined for the origin of pulmonary arteries; the morphology of the right ventricular outflow tracts and their relationship to the semilunar valves. Of 305 (93%) specimens of cardiovascular anomalies, 165 (54%) had PTA, 85 (28%) had HPT and 55 (18%) had isolated aortic arch anomalies. Using the classification of Collett & Edwards (1949), Type I, 12.1% (20/165); Type 11, 69.7% (115/165); Type 111, 12.7% (21/165); Type IV, 5.5% (9/165) were identified. The truncal valves were tricuspid in 106 (64%) and quadricuspid in 58 (35%) instances. The truncal valves overrode the VSD in all specimens of PTA and the dextroposition

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of the truncus was most frequent. There is no evidence of an infundibular septum in 165 PTA hearts. 71 hearts with HPT (or atresia) and VSD had dextroposition of the aorta that straddled the VSD and hypoplasia of the infundibular septum. The aortic valves were tricuspid in 95% and quadricuspid in 4% of the cases. It was concluded that the relationship between PTA and HPT (or atresia) represented morphologically discrete conditions.

Mortality rates in all experimental groups were found to be higher than f o r controls. Increasing the rates and duration of centrifugation induced higher mortality and more complicated cardiovascular anomalies. Cardiovascular anomalies were classified into two groups, one with and one without gastroschisis. Double outlet right ventricle, hypoplatic right ventricle, pulmonary stenosis and truncus arteriosus were also noted. Malformation syndrome induced by extremely high centrifugal force may be due to abnormal general fetal growth and abnormal development of the thoracic-abdomen system.

ISHIKAWA, S., N. KOMORI, M. NAGAO, H. OKAWA, T. SUGIUCHI and K. OKUYAMA. Department of Pediatrics, Showa University, Tokyo. Cardiovascular anomalies induced by vibration of the cardiac loop in chick embryos. The purpose of this study was to investigate the abnormal morphological development of the cardiac loop by direct stimulation using a micro-vibrating apparatus. Chick embryos at Hamburger-Hamilton stages 20 - 26 were used. A square opening was made in the egg shell to provide clear view of the embryo. Vibration at 50 to 100 cycles per minute was applied to the long (L) o r horizontal (H) axis of the cardiac loop. Both groups were examined for morphological differences. While stimulating the cardiac loop, a 16 mm movie was used to record abnormal development o f the cardiac loop. Embryos were sacrificed and necropsied after 17 days of incubation. Among survivors, 7/10 (70%) of the L group and 4 / 1 0 ( 4 0 % ) of the H group demonstrated cardiac malformation. The precise morphogenetic and geometric measurements revealed the elongation of the left ventricular inflow tract (10 mm/7 mm) in the L group and widening of the atrio-ventricular orifice (5-6 mm/4 mm) in the left ventricles of the H group. Complicated cono-truncal anomalies were observed in 7 / 1 0 (70%) of the L group and large ventricular septa1 defects were found in 4 / 1 0 ( 4 0 % ) of the H group. In the control group, cardiovascular anomalies were not found. Direct vibration of the cardiac loop distorted the basic morphological development of the primitive cardiovascular systen and induced complicated cardiovascular anomalies.

NAGAO, M., N. KOMORI, S. ISHIKAWA, and K. OKUYAMA. DeDartment of Pediatrics. Showa University, Tokyo. Spectrum of cardiovascular and extra-cardiovascular anomalies induced by ultra-centrifugal forces. The ultra-centrifugal force was used to induce cardiovascular anomalies and extracardiac malformations in chick embryos. An automatic high speed refrigerated centrifuge, 20 PR-52-type, was used to centrifuge fertilized eggs, warmed to 30C and set in 500 pp tubes around the RP RS 3-3 swing rotor. Fertilized eggs were subjected to centrifugation at 500 rpm for 5 o r 10 minutes OK 1000 rpm for 5 or 10 minutes.

MIYATA, K., E. YAMAMOTO, T. TAKESUE, S. ONO, M. NINOMIYA and S. MUNAKATA, Department of Pediatrics, Faculty of Medicine, Kagoshima University, Kagoshima. The production of cardiovascular malformations using antikidney and antiheart antibodies. In order to produce cardiovascular malformations in rat embryos, the following experiments were undertaken using tissue antibodies. The sera, kidneys and hear-ts of Wistar rats were removed, homogenized and injected S.C. with Freund's adjuvant into albino rabbits. The activity of the antisera was tested by a tanned red blood cell agglutination method. Pregnant rats of the same strain were divided into 5 groups according to the following method of administration. Group A was injected i.p. with various doses (0.15 - 0.45 m1/100 g) of antikidney sera (AKS) on day 9 . Group 3 was injected with small doses ( 0 . 1 0 m1/100 g) of AKS on day 9. Group C was injected with various doses (0.15 - 0.90 ml/100 g) of antiheart sera (AHS) on day 9 . Croup D was injected with small doses of AKS on day 9, and various doses of AHS on day 10. Croup E (control) was injected with various doses of antiratserum sera on day 9 . Treated mothers were sacrificed on day 21, and fetuses were removed. Incidences of malformations were 76.5% (78/ 102) in group A, 3.0% ( 2 / 6 6 ) i n group B, 3.2% (6/187) in group C, 3 2 . 8 % (88/268) in group D, and 2.7% (5/183) in group E. The malformations mainly observed in group A were central nervous system defects, and in group D were cardiovascular defects, such as VSD, double outlet of the right ventricle, ASD and double aortic arch. The results suggest that antiheart sera alone will not be teratogenic, but when given with subteratogenic doses of antikidney sera they will become teratogenic.

SINGH, S., S. SHAMAL and S. SINGH, Department of Anatomy and Ophthalmology, Institute of Medical Sciences, Banaras Hindu University, Varanasi, India. Retinal changes in chick embryos induced by excess zinc. Effect of excess zinc in developing chick embryos has not been studied. Single doses of

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zinc sulfate i.e. 0.8, 2.4 and 4.0 mg/egg were injected into the yolk sac of White Leghorn chick embryos on the 3rd, 4th and 5th day of incubation. Controls received equal volumes of distilled water. Whole eyes were dissected after the embryos were collected on day 1 9 . Light microscopic examinations revealed a hyperpigmented iris epithelium, a thick retinal pigment layer, a loss of protoplasmic process of rods and cones and poor differentiation of the ganglionic cell layer. Hyperpigmentation was found in all the treated groups but other retinal changes were seen only in the highest dose group treated on the 3rd or 4th day of incubation. Excess zinc enhances the pigment inductive factor and delays other components of developing chick eyes during organogenesis.

was observed. SEC consisted mostly of immature glial and neuronal cells and some macrophages. In the retina o f postnatal rats, SEC was seen 24 hours after treatment. The occurrence of SEC in the developing retina of the treated group may be regarded as a phenomenon of tissue repair.

SHAMAL, S., S. SINGH and S . SINGH, Department of Anatomy and Ophthalmology, Institute of Medical Sciences, Banaras Hindu University, Varanasi, India. Postnatal effects in rat offspring receiving adriblastina. Adriblastina, a new anticancer antibiotic is used for the treatment of a wide variety of cancers. A single maximum therapeutic dose of adriblastina i.e. 2.4 mg/kg was injected (I?) into rat offspring on post-conception (PC) day 22. Morphological and morphometric data were recorded daily up to PC day 43. Out of 2 4 treated rats, 33% died within this period. Dorsal to ventral position, forward movement, hair appearance, opening of pinna and eyelid opening were delayed for 1-3 days compared to controls. Fifty per cent of the treated offspring showed bilateral cataract. In 8% of the cases there was no opening of the eyelid on the left side. All the dead rats showed cataract on postmortem examination. The inhibition of DNA synthesis by the drug is assumed to be responsible for the postnatal mortality, retarded growth and induction of malformations.

NAGAISHI, T., T. MATSU0,T. IKEDA and I. TAKAKU, First Department of Pathology, and Department of Ophthalmology, Nagasaki University School of Medicine, Nagasaki.. Supraependymal cell clusters induced by ENU treatment in the developing rat retina. Supraependymal cell clusters (SEC), consisting of neurons, glial cells and macrophages, have been observed in the ventricular cavity of the brain in several mammalian species. A recent report suggested that SRC may be related to the pathologic structure of the central nervous system. The present study examined SEC in the rat retina induced by ENU treatment on the 14th day of gestation and on the 3rd day after birth. I n the fetal retina, SEC first appeared in the optic ventricle 12 hours after treatment with ENU. Subsequently, massive necrosis of the matrix layer, destruction of the ventricular surface and protrusion of the presumptive ganglion cell layer into the optic ventricle

TAKEUCHI, Y. K., H. SAKAI, M. KAWABATA and I. K. TAKEUCHI, Department of Anatomy, Nagoya University School of Medicine, Nagoya, and Institute for Developmental Research, Aichi Prefectural Colony, Kasugai, Aichi. Congenital aplasia of the optic nerve in microphthalmic rats induced by prenatal X-irradiation. Congenital hypoplasia of the optic nerve has been observed to be associated with congenital microphthalmia induced by several teratogens in human and experimental animals. However, only a few reports have been published on congenital aplasia of the optic nerve. We induced uni- o r bilateral microphthalmic offspring associated with congenital aplasia of the optic nerve delivering a single dose exposure of 100 R of X-rays to pregnant MP1 albino rats on gestational day 10. After rearing for two months, 15 microphthalmic eves were excised and examined microscopically. In 12 of these eyes, the nerve fiber layer, optic papilla and retinal vessels were absent, and the cell number in the ganglion cell layer was significantly decreased. In one eye, the neural retina had the anomalies largely similar to the above-mentioned eyes, but there was a small ectopic bundle of nerve fibers, which might be a hypoplastic optic papilla. The remaining two eyes had a nearly normal nerve fiber layer, somewhat distorted optic papilla, and normalappearing retinal vessels. I n the optic papilla, a large mass of irregularly accumulated nerve fibers was present. In embryological studies to clarify the cause of aplasia of the optic nerve, it was shown that the optic stalk was very thin in the embryo on gestational day 15 (120 h after Xirradiation), and thereafter, gradually disintegrated from the central area to the bilateral centrifugal direction. (Supported by Scientific Research Grant No. 56770788 from the Ministry of Education of Japan.)

SREENATHAN, R. N., R. PADMANABHAN and S. SHIGH, Department of Anatomy, Institute of Medical Sciences, Banaras Hindu Univerity, Varanasi, India. Comparison of teratogenicity of acetaldehyde and ethanol in the rat. Very low doses of ethanol (EtOH) (75, 200 and 300 mg/kg) and acetaldehyde (AcH) (50, 75, 100 and 150 mg/kg) were administered (IP) separately to pregnant CF rats from day 8 to 15 ) . Fetuses of gestation (sperm positive = day 1 collected at term showed a significant reduction in CR length, tail length and trans-

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umbilical distance. Gross malformations included microcephaly, micrognathia, micromelia digital anomalies, hemorrhages, skeletal defects etc., which are very much in keeping with the pattern of anomalies associated with fetal alcohol syndrome (FAS). Pair-fed and vehicle treated controls employed in this study rule out extraneous variables. AcH, the primary metabolite of EtOH induced malformations and growth retardation similar to those resulting from EtOH treatment. This implicates AcH in FAS.

PADMANABHAN, R., R. N. SREENATHAN and S. SINGH, Department of Anatomy, Institute of Medical Sciences, Banaras Hindu University, Varanasi, India. Histopathological changes of the placenta following treatment with ethanol and acetaldehyde in the rat. Eight doses of ethanol (EtOH) (75, 200 and 300 mglkg) and single and triple doses of acetaldehyde (AcH) (50, 75 and 100 mg/kg) were given intraperitoneally during the organogenetic period to two separate groups of CF rats. At term, fetuses of both groups showed malformations, similar to the abnormalities seen with the fetal alcohol syndrome. Placentae were generally reduced in weight and contained white necrotic rims. The decidua basalis showed invasion of inflammatory cells. Though multinucleate giant cells and large cysts containing acidophilic substances were found in the basal zones in both groups, AcH placentae had a richer aggregation of glycogen cells whereas EtOH placentae showed a depletion of glycogen. The labyrinthine zone showed extensive hyalinisation, perivascular fibrosis, a thickened placental barrier and reduced arborisation of fetal vasculature. These changes indicate that both AcH and EtOH are deleterious to the placenta, though the exact mechanism is yet to be clarified.

evident, i.e., growth retardation of the palate and delay in movement of the palatine shelves from a vertical to horizontal position were noted. The range of stages of palatal development observed on day 15 of gestation was wide ( 2 -7) in the case of LA-treated embryos, while the range was restricted to the stages before the beginning of shelf fusion (2 - 4 ) in the case of TA-treated embryos. Scanning electron microscopic studies of the palatal shelves revealed that the same epithelial changes characterized by loss o f microvilli and intercellular gaps noted in the region of presumptive fusion of normal embryos were found in embryos of both treated groups at the corresponding stage in palate development and the area of the shelf. Since these surface alteration have been considered to be necessary for the normal palatal fusion, it is suggested that the potential to fuse is not altered by these treatments.

KAMIMURA, M., T. KIHARA, T. MATSUO, J. HANDA, and T. TANIMURA, Department of Anatomy, Kinki University School of Medicine, Sayama-cho,

in LA-exposed and TA-exposed mouse embryos were compared. On day 11 of gestation, Jc1:ICR mice 0 0 mg/kg LA i.v. or 10 mg/kg were treated with 1 TA i.m. The morphogenetic stages of palatal development (-1-7) were determined by the method of Biddle (1980). On day 18 of gestation, cleft palate was present in 38.2% of the LA-treated fetuses and in nearly 100% of the TA-treated fetuses. In both treated groups, higher incidences of embryonic death, smaller length of live embryos and delay of their palatal development were observed from day 14 of gestation compared to normal fetuses. On day 15 of gestation, the delay of palatal development in relation to the CR length was

SATOH, H. and S. SHIMAI, Department of Hygiene Fukushima Medical School, Fukushima. Brain retention of mercury in mice prenatally treated with methylmercury and selenite. The protective effects of selenite against methylmercury toxicity in adult animals have been shown in many experiments. However, little is known about the modifying effects of selenite on methylmercury toxicology in fetuses and neonates. We describe mercury retention i n the brain, which is considered to be a target organ, of pups born from pregnant mice treated with methylmercury and selenite. Groups of pregnant mice were injected S.C. with 30 umol/kg of either methylmercury chloride (MeHg) or sodium selenite (Se) in saline on day 9 of gestation. Another group of mice were injected with a combination of methylA control mercury and selenite (MeHg+Se). On group consisted of NaCl injected mice ( C ) . the day of birth (D-I), the third day (D-3) and the eighth day (D-8) after birth, two pups from each group were sacrificed. Organs were collected and analyzed for mercury by the combustion method using a gold trap. Mercury concentrations in brains of the C and Se groups were negligible. In the MeHg group, mercury concentrations were 6.7, 3.7 and 1.1 ug/g for D-1, D-3 and D-8, respectively. In MeHg+Se group, concentrations were slightly lower, 5.3, 2.8 and 0.8 pg/g, respectively. Statistical differences were found between these two groups on D-3 and D-8. The amounts of mercury in the whole brain decreased slowly with no statistical difference between D-1 and D-3 of either MeHg or MeHg+Se groups. This suggested elimination of mercury from the brain is slow in neonates.

. WATANABE and A. ENDO, Department SATO, F., T of Hygiene and Preventive Medicine, Yamagata University School of Medicine, Yamagata. Cytogenetic effects of zinc deficiency on oogenesis and spermatogenesis.

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In a series o f studies on the effects of zinc deficiency on mammalian reproduction, cytogenetic effects on oogenesis and spermatogenesis were examined. Male and female mice of the ICR strain were fed either with a severely zinc deficient diet (0.5 ppm Zn), a marginally zinc deficient diet (10 ppm Zn) o r a control diet ( 5 0 ppm Zn) ad libitum. Female mice were mated with vasectomized male mice after 8 days of feeding. Mice with vaginal plugs were killed and oocytes were flushed out of the Fallopian tubes. Recovered oocytes were examined morphologically and cytogenetically (M 1 1 ) . Male mice were killed after 4 weeks of feeding, and chromosomes of spermatogonia (mitosis) and spermatocytes (M I, M 11) were examined. In the 0 . 5 ppm Zn diet group, the proportions of degenerated oocytes and oocytes at anaphase I stage were increased as compared with the 10 ppm and 50 ppm Zn diet groups. The Zn deficient groups had a tendency to have more aneuploid oocytes than controls. However, there was no increase in the incidence of chromosome aberrations in spermatogonia or spermatocytes in the zinc deficient groups. These findings suggest that zinc deficiency is not only teratogenic, as we have previously reported, but also has the deleterious effects on maturation processes of oocytes. Further, oocytes seem to be more sensitive to zinc deficiencies than spermatogonia or spermatocytes.

reflex score was shown in the MeHg group as compared with MeHg+Se on Day 1. These findings were consistent with the findings in adult animals, and suggest the possibility that selenite has a modifying effect on methylmercury toxicity in fetuses and neonates.

SHIMAI, S. and H. SATOH, Department of Hygiene, Fukushima Medical School, Fukushima. Behavioral development of mice prenatally treated with methylmercury and selenite. It has been shown that behavioral changes can be induced in offspring prenatally exposed to methylmercury. Selenite has protective effects against methylmercury toxicity in adult animals. However, little is known about the modifying effects of selenite on methylmercury toxicology in fetuses and neonates. The present experiment investigates the effects on development of behavior in offspring from pregnant mice treated with methylmercury and selenite. Pregnant CFW mice were injected S.C. with 30 pol/kg of either methylmercury (MeHg) or sodium selenite (Se) in saline on day 9 of gestation. Another group of mice were injected with a combination of methylmercury and selenite (MeHg+Se). Controls were injected with On the day of birth, the saline only (C). third day and the eighth day after birth, the behavioral development of eight pups in each group were estimated during 3-min observations. The behavioral test was a modified method described by Bolles and Woods ( 1 9 6 4 ) . The occurrences of twitching, head moving, righting reflexes, locomotive activities and grooming behavior were checked. In general, there was no vast difference among the behavioral development in each group. A slight tendency of behavioral retardation was shown in the MeHg group on Day 1 and Day 3. A significant deterioration in the righting

SINGH, J. D., Anatomy Department, Medical Faculty, Malaya University, Kuala Lumpur, Malaysia. Vidarabine induced birth defects in mice. Vidarabine, an anti-viral drug in clinical use i s reported to be teratogenic in rats and rabbits (Schardein et al, Teratology, 1 9 7 7 ) . The present communication reports the findings in mice. The day on which sperm was seen in a vaginal smear of females was designated as day 0 of gestation. Vidarabine was injected intramuscularly, intravenously or intraperitoneally to mice on days 7 through 14 of gestation in a dose range of 50 mg to 200 mglkg of body weight. The vehicle was water, and the injection volume was 1 ml per mouse. Controls received only water by the same routes at the same stages of gestation as the drug treated mice. The fetuses were collected by uterotomy on day 18 of gestation and were examined for external anomalies before fixing in 10% neutral formalin. Fetuses were dissected to study visceral anomalies and processed with alizarin red for staining of the bones. In addition to growth retardation, defects o f cranium, limb bones and viscerae were observed. In addition to species differences (when compared with previous findings in rats and rabbits) conspicuous, route specificities were noted in the same strain of mice. The details are discussed.

FUKUI, Y., I. HAYASAKA, M. INOUYE and Y. KAMEYAMA, Research Institute of Environmental Medicine, Nagoya University, Nagoya, Aichi. Microcephaly induced in mice by prenatal exposure to ochratoxin A. The relationship between the nature of developmental disorders of the cerebral cortex and the developmental stage at which the embryos and fetuses were exposed to ochratoxin A was investigated in offspring of pregnant mice. S1c:ICR female mice were injected intraperitoneally with a single dose from 3 t o 10 mg/kg between day 7 and 14 of gestation. Early pathological changes of the embryonic and fetal neural tissue were investigated 6 to 72 hours after injection, and the brains were examined for malformations at near-term and six weeks after birth. Ochratoxin A had a potent lethal effect on undifferentiated proliferating cells in embryonic brains. This effect was marked when the treatment was made before day 11 of gestation. With treatment on day 9 or 10, the cell death in the embryonic brain occurred 2 4 - 3 6

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hours after drug administration. Cell death was delayed compared to that caused by Xirradiation and other teratogenic chemicals administered on the same gestational days. In near-term fetuses, exencephaly, severe microcephaly with defects of the olfactory bulb and severe microcephaly were observed at high frequencies when exposed to ochratoxin A on day 7 - 8, day 9 or day 10 of gestation, respectively. The sensitive period for microcephaly was the first half of the organogenetic period before day 11 of gestation. Little change resulted from treatments after day 1 3 . (Supported by Scientific Research Grants, Nos. 56030047 and 56122018 from the Ministry of Education, Japan.)

. and Y. KAMEYAMA, Research InstiHAYASAKA, I tute of Environmental Medicine, Nagoya University, Nagoya, Aichi. Early pathological changes of ochratoxin A-induced exencephaly in mice. Ochratoxin A (OA) has been shown to produce CNS malformations in mice. When administered to pregnant mice on day 7 of gestation, OA induced exencephaly with a high frequency. The present study was performed to trace the series of pathogenetic events at early stages of exencephaly. OA at 4 mglkg was injected ip into pregnant S1c:ICR mice on day 7 of gestation (plug ) . Pregnant females were killed at day=day 0 various intervals from 2 to 72 hours after OA treatment and their embryos were examined microscopically. The first pathological changes observed 6 hours after treatment were pyknosis and inclusions in the neuroepithelial cells of the cephalic region. These changes spread throughout the caudal region of the embryo with time after OA treatment, and the entire neuronal region evidenced tissue damage at 16-20 hours. The severely affected embryos showed cytoplasmic vacuoles and disruption of the neuroepithelial structure. Damaged cells decreased in number at 24-28 hours and almost disappeared at 36 hours. Similar changes could also be observed in the surface ectoderm and mesoderm 16-20 hours after treatment but were much less severe than in the neuroepithelium. The above results suggest that the primary change leading to OA-Induced exencephaly was degeneration of the neuroepithelium. (Supported by the Scientific Research Grants, Nos. 56122018 and 56030047 from the Ministry of Education, Science and Culture, Japan.)

In this experiment, the role of damage in the matrix layer and radial fibre on the pathogenesis of cytoarchitectural abnormalities in the cerebral cortex was examined. When pregnant mice were injected with 50 mgl kg body weight of cytosine arabinoside (Ara-C) at 14 days of gestation and the embryos examined 3 hours later, many pyknotic nuclei were found restrjcted to the matrix layer surrounding the lateral ventricles. Twenty four hours later, nuclear debris was concentrated in the matrix layer. Scanning electron microscopic observation of the degenerating matrix cells first demonstrated numerous small hollows on the cell surface. 1 2 hours after injection, degenerating cell surfaces were honey combed and some cells were being phagocytosed by macrophages. When pregnant mice were injected with 30 mglkg body weight of Ara-C on days 13.5 and 14.5 of gestation and embryos examined 24 hours after the last injection using SEM, the ventricular surface was severely destructed and the matrix cells were exposed to the ventricles. Arrangement of radial fibers was disturbed and the radial arrangement of the young neurons in the cortical plate was absent. (Supported by Grant No. 81-11-03 from NCNMMD of the Ministry of Health and Welfare, Japan.)

ABE, Y . , Y . YAMANO, S . OHTA, and M. SHIMADA, Department of Pediatrics, Shiga Medical College, Otsu, Shiga. Light and electron microscopic obsenrations of the abnormal matrix layer in mouse embryonic brain. Histopathogenesis of cytoarchitectural abnormalities, often found in the brain of severely handicapped children, remains unknown.

YAMANO, T., Y. ABE, S. OHTA, T. UTSUNOMIYA, M. OHNO and M. SHIMADA, Department of Pediatrics, Shiga University of Medical Science, Otsu, Shiga. Morphological study of Purkinje cell dendritic spines after neonatal administration of cytosine arabinoside. Golgi studies have shown that spines of neuronal dendrites decreased in number and were malformed when developing brains were damaged by various agents. This study was undertaken t o elucidate the pathogenesis of this spine dysgenesis. ICR-JCI. mice were used for this study. The day of birth was counted as day 1. The mice, which were injected with 30 mg/kg body weight of cytosine arabinoside on days 3 , 4 and 5 , o r on days 7, 8 and 9, were designated as group I and group 11, respectively. Mice injected with saline solutions on days 2 , 3 and 4 served as controls. Mice in each group were killed at 60 days of age, and the spines of Purkinje cell dendrites were examined by electron microscopy. In the control mice, Purkinje cell dendritic spines made synapses with parallel fibers in the molecular layer. Mice from groups I and I1 frequently had unattached spines which did not make synapses in the molecular layer. Granule cells were decreased in number by administrations of cytosine arabinoside. Spines which established synapses were measured for profile size and synaptic length and quantitatively analyzed by computer. The average size was 0.17~0.08~m2 ( n = 1 6 4 ) in group I, 0 . 1 4 k 0 . 0 7 pm2 ( n = 1 6 0 ) in group I1 and 0.11+0.05 pm2 ( n = 2 6 6 ) in controls. The average synaptic length was 0.4010.14 urn ( n = 1 6 4 ) in group I, 0.34kO.13 pm (n=160) in group I1 and 0.34+0.11 pm ( n = 2 6 6 ) in controls. The results demonstrate that the Purkinje cell dendritic spines, able to make a synapse with a few parallel fibers, were larger than control cells.

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EGUCHI, M., S. MATSUMOTO, E. SHIOTA,

T. YAMAGUCHI, K. SHIBATA, H. KAWAMURA,

T. SHIMAUCHI and S. HANDA, Department of Orthopaedic Surgery, and Institute for Animal Experiment, Faculty of Medicine, Kyushu Univer-

decreased in number. Some PdnlPdn fetuses were involved with preaxial oligodactyly. These results indicate that 5-FU and Ara-C has an enhanced effect on the morphogenesis of polydactyly in Pdn/+ and an inhibitory effect on Pdnlpdn.

performed, detailed data on a diaplacental transfer of EHDP and the effect of EHDP on skeletal development are scarce. In this study, we demonstrate that the administration of high doses of EHDP to pregnant mice results in calcification and ossification deficiencies of the cartilaginous anlage resulting in angulation deformities of the long bones in mouse fetuses. EHDP, at 200 mglkg, was administered subcutaneously to pregnant mice on day 10, 11, 12, 13, 14, 15, or 16 of pregnancy. The animals 8 and fetuses were were sacrificed on day 1 removed surgically. Fetal skeletons were examined in alcian blue and alizarin red S stained preparations gross anatomically, and in sections histologically and histochemically. Administration o f EHDP on day 1 2 , 13, or 14 of pregnancy produced shortening of the mineralization zone, a widening of the columnar cartilage zone and angular deformities i n the long bones. Alkaline phosphatase activity was similar in non-mineralized and mineralized bone and cartilage. These findings show that EHDP can inhibit calcification without inhibiting the activity of alkaline phosphatase.

NARUSE, I. and Y. KAMEYAMA, Research Institute of Environmental Medicine, Nagoya University, Nagoya, Aichi. Digital malformations caused by 5-fluorouracil and cytosine arabinoside in genetic polydactyly mouse (Polydactyly Nagoya, Pdn) Pdnlpdn fetuses show preaxial polydactyly o f duplicated or triplicated metacarpal/metatarsal type in the fore- and hindlimbs. pdnl+ offspring have one extra-digit preaxially in the hindlimb. It is well known that in mice and rats 5-fluorouracil (5-FU) or cytosine arabinoside (Ara-C) induce malformations of digits including preaxial polydactyly. Pdn/+ females mated withPdnl+ males were treated with 20 mg/kg of 5-FU or 5 mglkg of Ara-C on day 10 or 1 1 of gestation. Abnormalities of the limbs of the fetuses were examined in bone and cartilage stained specimens on day 18 of gestation. In P d n / + or +I+, polydactyly of proximal phalangeal type on the second digital ray and preaxial polydactyly of metacarpallmetatarsal type with occasional tibial hemimelia were observed when treated with 5-FU or Ara-C on day 10. In PdnlPdn, the number of extra digits decreased and tibial hemimelia became severe when treated with 5-FU o r Ara-C also on day 10. When treated with 5-FU or Ara-C on day 11, postaxial oligodactyly in the fore- and hindlimb was observed inPdnl+ or +/+. Preaxial oligodactyly was also observed in some cases. In Pdnlpdn, extra digits of the preaxial side

SHOJI, R., A. HANAI, F. SUZUKI and R. YASUMURA, Department of Embryology, Institute for Developmental Research, Aichi Prefectural Colony, Kasugai, Aichi. A dominant mutation causing maldevelopment of axial skeleton in mice. A male mouse with a very short and kinky tail was found in an inbred strain bearing a recessive cataract gene cts. The mouse was crossed with females from the same strain and produced several descendants with the same eccentric character. To determine the heredity of this abnormal character, cross experiments were carried out between the abnormal descendants and normal mice from inbred KYF/MsIdr strain. The abnormal phenotype occurred in about half of the FI descendants, and crossbreeding within the FI descendants resulted in abnormalities; 0% in normal x normal, 4 9 % in normal x abnormal and 75.5% in abnormal x abnormal. The findings revealed that the abnormal character was due to a single autosomal dominant gene. The gross anatomy of the abnormal mice was studied in alizarin clearance specimens. The most striking departures in comparison with normal were found in the sxial skeleton. The vertebral elements were reduced in number, abnormally formed, irregularly fragmented and fused with each other. The spine often showed kyphosis, lordosis and scoliosis. It was difficult to determine the exact number of vertebrae. Often as many as two to four ribs were abnormally fused together with large spaces between the groups. The skull and the extremities were usually normal. In embryological observations, the abnormal fetuses were recognizable externally by their shortened tails down to 14 days of gestation. The tails of 13day abnormal fetuses were indistinguishable from the normal length, but their vertebral primordia displayed irregular shape and size.

ISHIBASHI, M., N. ODAGIRI, S . TAKATSU, T. SATOH and H. HARAGUCHI, Department of Animal Breeding, School of Veterinary Medical Sciences, Azabu University, Sagamihara, Kanagawa. sensitivity of Ishibashi Rats (ISR) to the teratogenic action of trypan blue. A new strain of rats with congenital spontaneous spinal abnormalities, especially of lumbar vertebrae, Ishibashi Rats (ISR), were used to study spinal abnormalities induced by trypan blue. A 1%aqueous solution of trypan blue (TB: Merck, C.I.Nr. 23850) was given intraperitoneally to nulliparous ISR rats on day 8 of pregnancy. On day 20, rats were anesthesized and fetuses surgically removed for examination. Fetal osseous and cartilagenous

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skeletons were prepared and stained by Inouye's method with alizarin red S and alcian blue. ISR treated with TB 40 mg/kg showed an increased post implantation l o s s of embryos and fetuses compared to controls. The skeletal specimens showed more frequent and serious abnormalities such as cleft, fusion and absent lumbar and sacral vertebral bodies and arches than controls. No dams were killed by TB 0 mglkg produced treatment. TB treatment at 1 slight spinal abnormalities of vertebral bodies and arches, especially of sacral and coccygeal vertebrae. From these results, ISR appears to be highly sensitive to spinal abnormalities produced by teratogenes such as TB. (Supported by a Grant in Aid for Scientific Research from the Ministry of Education, Sciences and Culture, Japan, No. 340009.)

MIKAMI, T., Y. MATSUBARA, H. YAMANAKA and Y. SUZUKI, Laboratories of Toxicology and Experimental Biology Research, Eisai Co., Ltd., Hashima-gun, Gifu. Teratogenic effects of thalidomide in rabbits: ( 3 ) The developmental processes of dysraphic malformations in JWNIRS rabbits. Previously, we reported that thalidomide induced a high incidence of craniofacial m a l formations such as anencephaly, holoprosencephaly and hydrocephaly, when administered to JW-NIBS rabbits (obtained from Nippon Institute for Biological Science) during organogenesis. We found that the telencephalic walls were everted at the midfrontal line in 11 day embryos in the group treated with thalidomide on days 6-9 of gestation, and the eversion became marked with advanced age. In this report, the developmental processes of the dysraphic malformations were examined in early embroys (222-264 hours postcoitus) from JW-NIBS rabbits treated with thalidomide (500 mglkg) at 168 hours after copulation. Under the dissection microscope, the anterior neuropore both in control and treated groups opened at 222 hours and closed in most embryos at 231 hours. At 243 hours, smallness of head, depression of the rhombencephalic portion and a slit-like reopening of the midfrontal neural tube were detected in the treated group. At 264 hours, the telencephalic walls were everted at the midfrontal line. In the histological sections, the significant changes at 231 hours were incomplete closure of the neural tube wjth closure of surface ectoderm at the midfrontal portion and sparsity of mesencymal tissue around the telencephalic region.

germ anomalies associated with jaw malformations in mouse fetuses caused by maternal hypervitaminosis A. This study was designed to investigate the morphological relationship between jaw malformations and developmental disorders of tooth germ in mouse fetuses following maternal hypervitaminosis A. Pregnant Jcl-ICR mice were given a single ip injection of 20000 IU vitamin A on day 8 , 9 or 10 of pregnancy (VP=O). On days 13-18, they were sacrificed, and the fetuses were examined for jaw and tooth germ abnormalities with bonestained, cleared specimens and histological sections. In the group treated on day 8, microstomiamicrognathia was produced with a high frequency and mandibles had severe reductional deformities. In the fetuses with a defect of the rear portion of the mandiblar corpus and ramus, incisor-like heterotopic tooth germs were observed in the hypoplastic maxilla and the remaining portion of the mandible. These abnormal tooth germs had osteodentine in their pulps, but no cusp formed. No tooth germs with molar structure were detected. In the fetuses with shortened corpus and without mandibular ramus, incisor-like heterotopic tooth germs were observed in the maxilla but not in the mandible. Molar germs were markedly hypoplastic. In the fetuses with only mildly deformed mandibles, no heterotopic tooth germs were observed and molar germs were fairly well developed. Significant changes in the devloping upper and lower jaws on days 13-14 were noted with excessive and heterotopic invaginations of the oral epithelium.

OKADA, S. Y., T. OKA, and Y. KAMEYAMA, Department of Oral Surgery, Faculty of Medicine and Research Institute of Environmental Medicine, Nagoya University, Nagoya, Aichi. Tooth

WATANABE, T., E. HOSHI, F. SAT0 and A . ENDO, Department of Hygiene and Preventive Medicine, Yamagata University School of Medicine, Yamagata. Teratogenicity of maternal biotin deficiency in mice. Biotin is an essential nutrient and plays an important role in mammalian reproduction. However, little is known of embryonic development in biotin deficient maternal mice. The teratogenic effects of biotin deficiency was studied by feeding pregnant mice with a diet of egg whites as the protein source. Pregnant mice fed on an egg white diet during the entire period of gestation produced a high frequency of fetuses with various kinds of external and skeletal malformations, with no apparent maternal effects. The predominant malformations produced were cleft palate (82%), micrognathia ( 9 1 % ) and micromelia ( 4 1 % ) . The mean weight of live fetuses and the number of ossification centers in the extremities were significantly decreased in the fetuses from biotin deficient mothers. All of these malformations were prevented by simultaneous supplementation with 10 mg/kg of biotin in the diet. These findings suggest that maternal biotin deficiency is highly teratogenic in mice.

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IWASE, T . . Y. HASHIMOTO, Y. MORIKAWA and M. MATSUO, Department of Veterinary Anatomy, College of Agriculture, University of Osaka Prefecture, Sakai, Osaka. Delayed closure of the rat secondary palate by 6-diazo-5-oxo-Lnorleucine (DON). The effect of DON, an antineoplastic agent, on the palatomorphogenesis and fetal development was studied in Wistar rats. Pregnant rats of the 14th day of gestation received intraperitoneal injection of DON dissolved into sterile phosphate buffer at a dose of 2 mglhead. Control rats were injected with sterile buffer. On the 16th day of gestation, fetal growth of rats exposed to DON was 6 to 12 hrs behind the development of controls, as evaluated by fetal body weight, CRL and growth of maxilla and mandible. Developmental delay was less during the 17th and 18th days of gestation. The elevation and adhesion of bilateral palatal processes occurred from 0 to 10 hrs on the 16th day of gestation in almost all controls. In DON exposed fetuses, the elevation and adhesion took place at a high frequency on the 17th day of gestation, and slow palatomorphogenesis followed. Histological features were normal in all other aspects. The delayed elevation of palatal processes is known to contribute in part to cleft palate. Nevertheless, the results of the present study suggest that, when the balance between palatomorphogenesis and orofacial formation is kept normal, even the delayed elevation, adhesion and fusion of bilateral palatal processes could lead to normal palatomorphogenesis.

NAKANE, K. and Y. KAMEYAMA. Research Institute of Environmental Medicine, Nagoya University, Nagoya, Aichi. Effect of maternal fasting on the manifestation of cleft lip in CL/Fr mice. Pregnant CL/Fr mice, which produce offspring with spontaneous cleft lip at a frequency of 24-26% at term, were fasted for 24 or 48 hours between days 7 and 11 of pregnancy (plug day=O). Animals were killed on day 18, and the frequency of intrauterine death and cleft lip of the living fetuses was investigated. Cleft lip occurrence and intrauterine mortality showed no significant changes in all groups fasted for 24 hours as compared to controls. In the groups fasted for 48 hours, however, the frequency of cleft lip in the fetuses from the mothers treated on days 8-9 (11 a.m. on day 8 to 11 a.m. on day 10) of pregnancy showed a significant decrease to 14% (nontreated control: 2 6 % , Wilcoxon's rank sum test, P <0.01). Intrauterine mortality in the same group increased to 21% (nontreated control: 15%). In the groups treated on days 7-8,'9-10, 10-11 and 11-12, intrauterine mortality increased (26-50%), but the cleft lip frequency failed to show any significant change (26-35%). Thus, the cleft lip frequency in CL/Fr mice was reduced by half when pregnant mice were fasted for 48 hours on days 8-9 of pregnancy. This reduction may he attributed to the relatively high mortality of the embryos with cleft lip. (Supported by the Takeda Scientific Foundation.)

ESAKI, K., Central Institute for Experimental Animals, Kawasaki, Kanagawa. Lower incidence of cleft lip by urethane administration in the Cl/Fr mouse. The effects of urethane on the incidence of cleft lip in the Cl/Fr mouse were examined by a single intraperitoneal administration at a dose of 100, 500 or 1500 mg/kg on day 8 , 9 or 10 of gestation (VP=O). On day 18 of gestation all mothers were sacrificed and their young were examined for the presence of cleft lip. The mortality o f embryos or fetuses in the 1500 mg/kg group was increased to 44-67%, hut those (12-17%) in the 100 and 500 mg/kg groups were comparable to the 14% incidence found in the control group. The incidence of cleft lip was decreased to 12% in the day 9-100 mg/kg group, to 3% in the day 9-500 mg/kg group and 0% in the Day 91500 mg/kg group, though that was 20% in the control group. The incidence of cleft lip in groups treated on day 8 or day 10 of gestation was comparable to that in the control group. This suggests that urethane administration on day 9 of gestation reduced the cleft lip incidence in Cl/Fr mice, and this reduction was not caused by a mortality increase of embryos with cleft lip. However, the mechanisms of this effect are not yet known.

YAMAMOTO, M., Y. TAKAYAMA and G. WATANABE, Department of Hygiene and Preventive Medicine, Niigata University School of Medicine, Niigata. Teratogenicity of antimouse testis antisera (AMTAS): Cross reactivity of AMTAS against kidney antigen and its fluorescent localization in maternal and fetal tissue. Previous studies in our laboratory have indicated that AMTAS was teratogenic when injected into pregnant mice on day 12.5. Administration of 17.8 mg and 35.6 mg of AMTAS (as total protein) produced 6.1% (7/58) and 27.3% (12/44) malformed embryos, respectively, while control malformation frequencies were 0% (0/161) and 0.6% (1/158). The crossreactivity of AMTAS was examined against testis extract (AgT), semen (AgS), kidney (AgK) and solubilized kidney (AgKs) using Ouchterlony's method. AMTAS formed three precipitin lines against AgT and AgS, and two lines against AgK and AgKs. One complete spur was observed between AgT and AgK. It was therefore suggested that a testis-specific antibody was present. Localization of AMTAS in embryos and uteri was examined by the immunofluorescent method. Flourescence was observed in the mesometrial vessels, trophectoderm and the Reichert's membrane. The amnion and the embryo, however, were less stained. It is therefore implied

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that the teratogenicity of AMTAS was not due to the direct effect on the fetus, but instead to the disturbed function of the placenta. Since the presence of a testis-specific antibody by Ouchterlony's method does not simply mean the presence of teratogenicity; the hypothesis in regard to the role of such an antibody needs further study. (Details of the AMTAS teratogenicity have been reported in Teratology, 24:53-63, 1981).

KANEDA, M., S. TERAMOTO and Y. SHIRASU, Toxicology Division, Institute of Environmental Toxicology, Kodaira, Tokyo. Pattern of anomalies induced by 1-methylthiourea in the rat fetus. 1-Methylthiourea (MTU) has been detected as one of the metabolites of ethylenethiourea (ETU) and has been reported to be teratogenic in the rat (Teratology, 23:335-342, 1981). When administered on day 12 o r 14 of pregnancy, the type of anomalies induced was similar to that obtained after administration of ETU. In the present experiment, a single dose of 200 o r 400 mg/kg of MTU was given orally to pregnant rats during days 6-14 of pregnancy in order to investigate the relationship between the type of anomalies and the time of treatment. The results were compared with that obtained in the ETU exposed rats. No malformations were observed with treatment during days 6-8 o f pregnancy. However, various types of anomalies were induced by administration of MTU during days 9-14 of pregnancy. The temporal relationship between the type of anomalies and the time of treatment was as follows. Tail anomalies (short or kinky tail) were produced in fetuses treated during days 9-14 of pregnancy. Treatment during days 10-14 of pregnancy induced anomalies in the urogenital organs including hypoplasia of the kidney, hydronephrosis, hydroureter, ectopic ureter, hypoplasia of the uterus, undescended testes and hypoplasia of the genital tubercle. Malformations in the CNS, such as meningocele and dilatation of the ventricular system, were observed after treatment during days 11-14 of pregnancy. Administration during days 12-13 of pregnancy resulted in anomalies in the extremities such as oligodactyly, syndactyly and hypoplasja of the long bones. These results were similar to those obtained after ETU treatment.

females, respectively. Furthermore, preimplantation embryos were karyotyped on days 2.5 and 4.5 of gestation with 23 and 14 diabetic females, respectively. The number of corpora lutea, embryos, and implantation sites were recorded. 1) There was no decrease in the number of corpora lutea, of collected embryos, o r of implantation sites in diabetic females. 2) As far as the preimplantation stages were concerned, there was no decrease in the cleavage activity and no increase in the incidence of developmental anomalies. 3) Postimplantation development, however, showed significant increases in embryonic death (P<O.OOl), gross malformation (P<O.OOl), and fetal hypoplasia (P<O.OO). 4) There was no increase of chromosomal aberrations in the preimplantation embryos. Results l ) , 2) and 4) indicate that in diabetic females, which were capable of conceiving during a normal estrous cycle, the maternal diabetic milieu did not affect oogenesis, ovulation, fertilization, cleavage division, or implantation. However, maternal diabetes exerted deleterious effects on the fetal development during the stage of organogenesis and reduced fetal growth during later developmental stages.

ETO, K . , F . TAKAKUBO and M. YONEYAMA, Section of Craniofacial Development and Anomalies, Institute of Stomatognathic Science, Faculty of Dentistry, Tokyo Medical and Dental University, Tokyo. Rat embryo culture during the period of organogenesis by an improved rotation apparatus. The rotation apparatus for rat whole embryo cultures is useful in experiments to test the effect of drugs on embryo organogenesis. Continuous movement of this apparatus ensures maximum and uniform exposure o f embryos to drugs. The original rotation apparatus was modified to allow a continuous gas supply to the embryos. The culture period of rat embryos explanted at gestational day 1 1 1/2 (plug = day 0) was about 48 hours in the old system and has been extended to 96 hours with the new culture method. Protein content and somite number of the embryos cultured in the modified technique also improved.

FIJNAKI. K. and K. MIKAMO. DeDartment of Biological Sciences, Asahikawa Medical College, Asahikawa, Hokkaido. Teratogenjc effects of maternal diabetes on embryonic development in a diabetic strain of Chinese hamsters. Females with typical diabetic symptoms were chosen from F13 - F17 of an inbred diabetic strain raised in our lat ratory. Embryological studies on days 2.5, 4.5, and 18.5 of gestation were performed with 23, 14, and 67 diabetic
~

- .

TAKAKURO, F . , M. YONEYAMA and K. ETO, Section of Craniofacial Development and Anomalies, Institute of Stomatognathic Science, Faculty of Dentistrv. , - Tokvo Medical and Dental Universitv. Tokyo. Induction of cleft lip by opening the yolk sac and amnion in cultured rat embryos. To study the role of the yolk sac and amnion in craniofacial development, the effects of opening the yolk sac and amnion on primary palate formation of rat embryo were examined in vitro. Formation of the primary palate in rat embryos takes place from day 11 1/2 to day
I

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13 1/2 of gestation. In experiments, the yolk sac and amnion were opened at the time of explantation (11 112 days) in one group (D11 Open) and were opened 24 hours after the beginning of the culture (12 1/2 days) in another group (D12 Open). The embryos of both groups were cultured for up to 72 hours. Cleft lip developed in almost all of the cultured embryos when the yolk sac and amnion were opened at day 11 1/2 (D11 Open). The embryos in group D12 Open were normal in primary palate formation. Protein content, wet weight and somite number of embryos were not significantly different in both groups. The results of this study demonstrate that opening the yolk sac and amnion at day 1 1 1/2 induces cleft lip in cultured rat embryos.

SEI, K., S . IIJIMA, Y. TACHIBANA and N. MATSUMOTO, Department of Public Health, Faculty of Medicine, University of Tokyo, Tokyo, and Department o f Health Sciences, Medical Universitv of Yamanashi. Tamaho. Yamanashi. Combined effects of cadmium and zinc on mouse embryo development in vitro. Schroeder et a1 reported the preventive effects of Zn with the simultaneous administration of the teratogen, Cd. We studied these effects using whole embryo culture. ICR mouse embryos ( 8 & 1 / 2 days, plug day = 1) were cultured in 4 ml human serum and 1 ml Waymouth's MB 752/1 medium with glucose (3 mg/ml) and containing various concentrations of Cd and/or Zn for 48 hours. Toxicity of Cd was assessed by somite number, external examination and protein content of the embryos. There was no significant difference in growth rates between control embryos and embryos exposed for 48 hrs to a range of CdC1, from lx10-6to Z X I O - ~ M. The effect of a pulse treatment of Cd and/or Zn was examined. At first, Cd was added to medium at 8:00 PM on the day when embryos were dissected and after 12 hours or 6 hours treatment of Cd, embryos were transferred to Cd free medium and cultured for 48 hours in total. Embryos treated with ~ X ~ O of ~Cd M were retarded but simultaneous treatment with Zn inhibited Cd retardation. Post-treatment of Zn had no effect on preventing retardation. Further investigations are directed at examining the effect of pulse treatments of Cd and Zn. (Supported by the Ministry of Education, Science and Culture, Japan)

with increased levels of salicylic acid (SA), a major metabolite of aspirin. Its effect was closely related to the decrease of protein and the increase of free fatty acid (FFA) levels in maternal rat serum. In this report, we investigated further, the effect of endotoxin on protein binding of SA in maternal and fetal serum, and obtained the following results. 1) Maternal protein levels decreased from 86 to 64 mglml and FFA levels increased from 0 . 4 up to 1.1 ueq/ml after injection of endotoxin (20 pg/kg, iv). These levels did not return to normal for eight hours. 2 ) The protein binding of SA was decreased according to the concentration of FFA and recovered by in vitro treatment of serum with activated charcoal. 3) No significant change was observed in the concentration patterns of maternal serum FFA with o r without endotoxin injection. 4 ) Protein and FFA concentrations in fetal serum were about one third of levels in maternal serum. These data suggest that the enhancement effect of endotoxin might be dependent on the change of the binding capacity of maternal serum proteins.

ITAMI, T. and S. KANOH, Department of Pharmacology, National Institute of Hygienic Sciences, Osaka. Mechanism of action of drugs on fetal toxicity: (VI) A possible role of protein binding in the enhancement effect of endotoxin on aspirin-induced fetal toxicity. Previously, we reported that enhanced fetal toxicity of aspirn by endotoxin was correlated

TANAKA, H., K. NAKAZAWA and M. ARIMA, Division of Child Neurology, National Center for Nervous, Mental and Muscular Disorders, Kodaira, Tokyo. Maternal caffeine and fetal development in rats. Daily caffeine ingestion during pregnancy has been suggested to increase the risk of congenital malformations, spontaneous abortion, stillbirth, premature delivery and low birth weight in humans. This study was undertaken to determine whether maternal caffeine ingestion was a risk factor in fetal development. Female Wistar rats were given water containing 0.04% caffeine 37 days prior to mating and remained in the diet for various times during pregnancy. Pregnancies were terminated on gestational day 20 by cesarean section. Litter size, fetal weight, viability and gross abnormalities were determined. Cerebral DNA, RNA and protein contents, and DNA synthesis in the cerebrum were measured as well as caffeine levels in maternal serum and fetal cerebrum. Maternal body weight and litter size were n o t significantly different in all groups. The survival rate of fetuses decreased in litters from dams ingested during pregnancy without pretreatment. There was no reduction of fetal body and total brain weight, although lower fetal cerebral weight was observed from dams ingested before mating and/or throughout pregnancy and for the last 6 gestational days. Thymidine incorporation into cerebral DNA was also reduced in litters from dams ingested throughout pregnancy. Maternal serum caffeine levels were positively correlated with fetal cerebral caffeine levels and negatively correlated with fetal survival rates. In conclusion, the adverse effects of maternal ingestion of 0.04% caffeine during pregnancy on rat fetus were seen in cerebral development and in fetal viavility.

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NISHIMURA, K., K. SATO, Y. TERADA, T. NANTO and K. YOSHIDA, Research Laboratories, Dainippon Pharmaceutical Co., Ltd., Suita, Osaka. Effects of indomethacin on the ductus arteriosus in rats. It has been reported that indomethacin treatment causes intrauterine constriction of the ductus arteriosus in fetal rats. However, a detailed study has never been performed. We studied the dose-response relationship of rat ductus arteriosus closure after single or multiple administrations of indomethacin. Indomethacin was administered by Ravage to 0 mg/kg 3, 6 or 24 pregnant rats at a dose of 1 hours before the observation of the ductus arteriosus at 3:OO p.m. on day 21 of gestation. Fetuses were delivered by cesarean section and fixed in 10% formalin. The diameter of the lumen of the ductus arteriosus was measured at the narrowest point with a dissecting microscope. Ductal constriction was observed 3 hours after treatment. The constriction reached its maximum 24 hours later. The inner diameter at 24 hours after treatment was 341212 and 21+18 pm for control and indomethacintreated group fetuses, respectively. Indomethacin treatment resulted in the constriction of the ductus arteriosus at the dose of I mg/kg that corresponded to daily human dosage level. On the other hand, no intrauterine ductal constriction was induced by treatment with 3 mglkg indomethacin daily from day 17 through day 20 of gestation. The mechanism in which the ductus arteriosus was not closed by repeated treatments is under investigation.

SUGIYAMA, O., K. I. TANAKA, M. OKAMOTO, S. IGARASHI, M. TSUKUSHI and S. SHIOTA, Research Laboratories 11, Chugai Pharmaceutical Co., Ltd., Tokyo. Effects of maternal hypervitaminosis-A on behavioral development of rat offspring. We have studied behavioral and functional abnormalities of offspring exposed pre- or perinatally to chemicals. In the present study, offspring with deformed ears were born to mothers exposed to vitamin A before parturition. The offspring were given various behavioral tests. Screening methods were described previously. We detected the features of the behavioral abnormality induced by vitamin A and determined the significance of the screening method. Wistar-Imamichi rats were exposed to 60000 USP/head of vitamin A, administered orally o r the 9th day of pregnancy. The tests consisted of ear-opening, teethcutting, eye-opening, righting reflex, negative geotaxis, rotary rod tests, auditory tests, open field tests, hot plate tests, water filled multiple T-maze tests, and shuttle box tests. Subjects were divided into two groups: subjects showing normal appearance (N-group) and subjects showing ear-malformation (Pgroup). The common behavioral change which was observed in both groups, was the high incidence of head-twitching in the open field test. The incidence rates were 29% in the N-group and 68% in the P-group, respectively. In addition, we observed an impediment in the auditory response in the P-group. Both changes may be the result of vitamin A toxicity on and around the acoustic organ.

YAMADA, T., H. UCHIDA, A. ICHIKAWA and M. SASAJIMA, Research Laboratories of Taisho Pharmaceutical Co., Ltd., Ohmiya, Saitama. Detection method for learning impairment of offspring in reproduction test. We examined learning and its retention using a water T-maze, passive avoidance response and a shuttle box in rats with impaired learning ability caused by prenatal treatment with N-methylnitrosourea (MNU) and chlorpromazine (CPZ) Wistar rats were treated with MNU ( 4 mg/kg, i.p.) on day 13 of pregnancy and CPZ ( 6 mg/kg, s.c.) on days 17-20 of pregnancv. The grade of detection obtained among the apparatuses employed in the learning test using the identical offspring was water T-maze > passive avoidance response > shuttle box in the MNU group, and shuttle box >passive avoidance response > water T-maze in the CPZ group. In the retention test, a passive avoidance response apparatus showed better sensitivity than other apparatuses in the detection of learning impairment of offspring in both MNU and CPZ groups. The results suggest that the retention test method, using passive avoidance response, is useful for evaluating the learning ability of offspring in the reproduction test.

KIHARA, T., Y. YAMAMOTO and T. TANIMURA, Devartment of Anatomv. Kinki Universitv School

treated prenatally with maternal sc injections with AFBl 0.25 mg/kg/day showed a significantly longer swimming time than controls in the underwater T-maze test, and a significantly lower avoidance rate in the acquisition of discriminated avoidance conditioning. The present sutdy was undertaken to examine the effects of AFB, on three behavioral parameters in postweaning offspring. Jc1:Wistar female rats were subcutaneously injected with 0 . 3 mg/kg/day of AFB, disolved in dimethyl sulfoxide for four days on days 1 1 - 1 4 (A) or ) . For 15 - 18 (B) of gestation (plug + = day 0 the postweaning test, male offspring born of the AFBi exposed mothers were tested on the rotarod ( 4 - 5 weeks of age), the open field test (5 - 6 weeks of age) and the avoidance In conditioning test (14 - 16 weeks of age). the rotarod test, the pups in A group remained on the drum at 15 rpm for a significantly shroter time on the second day than the control group. There were no significant treatment

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effects for any of the measures of the open field performance. In the acquisition of discriminated avoidance test, the learning rate from the 5th session was significantly lower in the offspring of A group. These results indicate that impaired locomotor coordination and learning deficiencies are induced in the offspring of mothers exposed to AFBi during pregnancy. (Supported by The Special Project on Environmental Science, Grant in Aid for Scientific Research, Ministry of Education.)

gait with a wide stance and broad angle of their hindlimbs compared to littermate controls.

TACHIBANA, T., Institute for Developmental Research, Aichi Prefectural Colony, Kasugai, Aichi. Validity of the open-field test in rats. __ 96 male rats were tested in an open field under dim-light or bright-light conditions for 3 min per day for 5 days. The validity of 7 open-field test measurements was assessed on the basis of the concept which was defined operationally using the test situation as the external criterion. Ambulation in the first 1 min and rearing showed not only validity but also some generality. An interactive measure (ambulation in the first min and defecation) was proposed as a new significant measure. Larger intercorrelations among measurements were obtained in a less stressful situation. A considerable constant factor-loading pattern across days and groups was noted, and this suggested the validity of the factor structure of an open-field test measurement. Scores obtained on day 1 showed some heterogeneity among scores obtained during 5 days.

TAMAKI, Y., S . ODA and Y. KAMEYAMA, Faculty of Education, Shinshu IJniversity, Nagano, and Research Institute of Environmental Medjcine, Nagoya, Aichi. The motor development of Rolling Mouse Nagoya (RMN). The Rolling Mouse Nagoya (RMN) is a mutant mouse strain, first described by Oda in 1969, is characterized by severe gait disturbances; hence it is regarded as an animal model of human cerebellar degenerative diseases. This study was undertaken to investigate the nature of the motor development of RMN. RMN young were obtained from mating of heterozygote ( C 3 H l t POL) ~ mice. Their motor development was observed between 9-22 days of age by recording various spontaneous activities, and were tested as follows: 1) bar holding, 2) rope descending, 3 ) narrow-path traversing, and 4 ) gait examination. In the rope descending test, RMN, showed more primitive modes of descent (fall and backward descent). Attempts at pull-ups with the forelimbs, and the synergistic support of the body with the hindlimbs were seldom seen in the bar holding test. Thus, RMN evidenced a severe lack of coordination and atonia of the hindlimbs. RMN also displayed a retardation of elevated quadruped locomotion and a wadding

. TANAKA, SUGIOKA, K., T. YOSHIOKA, and 0 Department of Anatomy, Shimane Medical University, Izumo, Shimane. Morphological changes and behavioral alterations in two-way avoidance tasks following neonatal hippocampal lesions in rats. This study was performed to determine the relationship between morphological changes and behavioral alterations in two-way avoidance learning following neonatal hippocampal lesions in rats. Neonatal Albino Wistar rats were randomly assigned to either a neonatal hippocampal lesion group (NHL group) or an intact group (Int group). NHL rats received bilateral anterodorsal hippocampal lesions with a radio frequency lesion generator at 1 day of age. I) Behavior: Rats were giJen a 3 min openfield test when they were 30 days old, and then trained in two-way avoidance exercises for 50 trials per day on 3 successive days from either the 31th OK the 61th day. During one trial, a 1 KHz tone CS, followed 10 sec later by 100 Vac OK 125 Vac US. 11) Morphology: The brains of NHL rats, at 5, 1 0 , 15, 20, 25, 35 and 65 days of age, were removed after perfusion with 10% formalin or an aldehyde mixture fixative. Tissues were stained by the KliiverBarrera method for light-microscopy or with E-PTA ( e t h a n o l i c - p h o s p h o t u n g s t i c acid) for electron-microscopic synapse morphometry. NHL rats at 30 days of age were more active in the open-field test. In the avoidance test using Lindquist's type VI ANOVA showed the main effect of groups, and groups by trials interaction and groups by days interaction were significant (pc.025, pc.01, pc.01, respectively). Avoidance learning was significantly facilitated in the NHL group. Analyses were made to reveal the relationship between the behavioral results and functional and morphological changes in the hippocampus.

MATSUO, T., J. HANDA, T. KIHARA, M. KAMIMURA and T. TANIMURA, Department of Anatomy, Kinki Universitv School of Medicine. Savama-cho. Osaka. Preweaning behavioral and functional development of spontaneously hypertensive rats (SHR) and stroke-prone SHR (SHRSP) rats. Spontaneously hypertensive rats (SHR) have been developed through selective inbreeding by Okamoto and Aoki from normotensive Wistar rats of a Kyoto strain (WKY). Stroke-prone SHR (SHRSP) were produced by selecting a substrain which was prone to develop cerebral hemorrhage. A comparison was made in the preweaning behavioral and functional development among SHR, SHRSP and M Y . The litter size was adjusted to eight on day 4 of age. The male and female offspring were
> ,

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examined for development of the surface-righting reflex on day 5. Latency between the supine and prone position was measured. Cliff avoidance was measured on day 6. A retraction from the edge within 10 seconds is considered to be successful. Negative geotaxis was assessed on day 7. Offspring were placed face down on a 25 degree plywood incline. Swimming development was scored on days 8, 10, 12 and 14. Direction, angle and limb usage were recorded. In physical development, SHR was the heaviest in body weight through the lactation period. SHR had a significantly shorter response time compared with SHRSP and WKY in surface-righting reflex. SHRSP showed a significantly shorter response time in the cliff avoidance and negative geotaxis among the three strains. In swimming, SHR gained a significantly higher score on direction and angle than SHRSP and WKY. However, the limb usage score was not significantly different among the three strains. These results suggest that straindifferences are present among SHR, SHRSP and WKY in preweaning behavioral and functional development.

SONTA, S., K. FLKLII and H. YAMAMURA, Department of Genetics, Institute for Developmental Research, Aichi Prefectural Colony, Kasugai, Aichi, and Department of Anatomy, Mie University School of Medicine, Tsu, Mie. Selective elimination of chromosomally unbalanced zygotes in the preimplantation period. The zygotic selection of genome imbalance was investigated by direct chromosome analyses of secondary spermatocgtes and preimplantation embryos in the heterozygous chromosomal system of the Chinese hamster [T(2;10)31dr (abbr.: T3)l. Our previous analyses of spermatocytes of T3/+ males and zygotes from the cross +/+p x T3/+& revealed that there was no evidence for selective elimination of chromosomally unbalanced gametes during the time from meiosis to fertilization. On the other hand, the frequency of 2-cell embryos karyotyped successfully decreased in the CKOSS + / + e x T3/+8 compared with the control mating (+I+?x and some karyotypes observed in 1-cell embryos from the same CKOSS were no longer seen after the 2-cell stage. These karyotypes had a deficiency of the chromosomes due to adjacent 2 or 3:l disjunctions of the quadrivalent. Both the rate of 2-cell embryos observed at the time corresponding to the 8-cell stage o f normal embryos and the increased frequency of preimplantation losses in the CKOSS +/+? x T3/+b were consistent with the expected frequency of embryos with karyotypes lost after the 2-cell stage. These findings show that embryos with karyotypes having a deficiency of the specific chromosomes stop their cleavage at the ?.-cell stage and hence are eliminated before implantation. Thus, the present results give the first cytological evidence for selective elimination of genome imbalance in early diplophase of the Chinese hamster.

FUKUI and M. SASAKI, Department of Anatomy, Mie University School of Medicine, Tsu, Mie. Temporal relationships of copulation with ovulation, sperm penetration, pronuclei formation and first cleavage in the ICR mouse. The relationships between times of copulation and those of ovulation, sperm penetration, pronuclei formation and first cleavage were investigated in ICR mice. Jc1:ICR mice housed in a room with a lighting schedule of 17 hrs light/7 hrs dark (lights on at 4 P.M.) were used. Females exhibiting external signs of estrus were placed monogamously with a vigorous male at 9 A.M. and when a copulation plug was found at one-hour intervals, females were separated from males. They were divided into 5 groups according to copulation time, i.e., A: 9-11 A.M., B: 11 A.M.-1 P.M. C: 1-3 P.M., D: 3-5 P.M., E: 5-9 P.M. Copulating females were sacrificed at various intervals, and the ovaries and ova were examined. Copulation rates in groups A - E were 41.6%, 2 6 . 3 % , 12.0%, 7.9% and 0.8%. respectively. Linear regressions with significant slopes (p<O.Ol) were obtained in groups A - D, when the arcsin-transformed proportions of ova ovulated, penetrated by sperm, with pronuclei or having finished the first Cleavage, were expressed in terms of hours (in group D a regression was calculated only for the first cleavage). From these regressions the times when each event occurred at a certain percentage of ova were estimated. Times when the same types of events in groups A - D occurred at the same percentage of ova were found to be earlier, almost in the order o f A - D. It was thought that sperm penetration in group C and the first cleavage in groups C and D occurred significantly later than in group A, because 95% confidence regions of regression lines did not overlap.

YAMAMURA, H., K .

+/+a),

INOUYE, M., S. ODA and Y. KAMEYAMA, Institute for Developmental Research, Aichi Prefectural Colony, Kasugai, Aichi, and Research Institute of Environmental Medicine, Nagoya University, Nagoya, Aichi. The early development of the house musk shrew, Suncus m r i n u s , a newly developed experimental animals. This report describes the morphology of the early development of the house musk shrew, Suncus m r i n u s , in comparison with tbat of other experimental animals. On day 7 of gestation the shrew blastocyst is implanted with its inner cell mass-side toward the uterine wall. The orientation is the same as in man but 1 8 0 ' different from that in a mouse. The type of implantation is eccentric, and the blastocyst comes to rest in the antimesometrial part of the uterus. The inner cell mass develops into three germ layers, without forming the egg cylinder characteristic o f the mouse and rat; instead, an embryonic disc similar to that in man is formed. On day 8 the heart rudiment is formed. The amniotic fold arises t o cover the dorsal s u r -

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face of the embryo and closes at the posterior end of the disc. The inner layer of the fold gives rise to the amnion and the outer layer eventually constitutes the clprion. These features were comparable to those in the rabbit. On day 9 the embryonic ectoderm develops into the neural plate. The first pair of somites appear. The embryonic disc grows to form head and tail folds. The allantois extends so as to be fused with the chorion, then penetrates the uterine crypts and forms the placenta in the antimesometrial part. This contrasts with the mouse and rat, whose placenta is formed in the mesometrial portion. Thus, the house musk shrew would appear to have several similarities with man in the morphologic features of early development, and it is suitable as an experimental animal for comparative teratological study.

INAMOTO, M. and H. YAMAMURA. Department of Anatomy, Mie University School of Medicine, Tsu, Mie. Morphological changes in the mouse uterine epithelium from copulation to implantation. Morphological changes in the endometrial epithelium from copulation to implantation were investigated in mice. Jc1:ICR mice housed in a room with a lighting schedule of 17 hrs light/7 hrs dark (lights on at 4 P.M.) were used. Females which copulated between 9 A.M. and 2 P.M. were perfused through the aorta with a fixative solution containing 2.5% glutaraldehyde and 2% paraformaldehyde at 6- to 24-hour intervals from 3 : 3 0 P.M. on the copulation day (day 0 of pregnancy). The endometrial epithelia in the antimesometrial side of the middle portion of the uterine horns were observed using light and electron microscopes. Intercellular leucocyte infiltration and autophagic vacuoles in the epithelial cells began to appear from 9 P.M. on day 0 to 3:30 A.M. on day 1 and became more intensive until 9:30 A.M. on day 1, but they disappeared by 9:30 on day 2 . At 9:3O A.M. on day 3, the epithelial cells had more abundant and regular microvilli than on day 2, and they contained many small vesicles in the apical region and a well developed Golgi apparatus. At 9:30 A.M. on day 4, the microvilli fused in part to become irregular and a few processes of the epithelial cells protruded into the uterine lumen. The apical vesicles were diminished in number and the Golgi apparatus showed inactive forms. By 9:30 A.M. on day 5 , the apical surfaces of the epithelial cells attached to each other in an interdigitational manner at other than implantation sites, and the apical vesicles were no longer found in these cells.

Medicine, Sagamihara, Kanagawa. Xenografting of rat embryonic organs into athymic (nude) mice, __ Because therapeutic grafting of human fetal tissues has recently been undertaken with few encouraging results, experimentally available related information is needed. As a continuation of our previous experiment along this line (Teratology 18, 152, 19781, we studied the ability of athymic nude mice (NFS) to accept Wistar rat embryonic organs and the histological differentiation of the readily accepted organs. Organs taken out from 16 to 21-day rat embryos were grafted into adult nude mice and were examined beginning one week later. Alike to the results with grafted human embryonic organs, rat intestines, lungs and limbs grafted subcutaneously, and retinal tissue grafted intratesticularly, were accepted nearly 100% of the time. The grafts of the small intestines from 16-day rat embryos developed into multilocular structures showing an indication of villi and epithelial differentiation from the original multilayered epithelium to the simple columnar epithelium. The grafted eyes contained retinal tissues arranged irregularly in the intertubular parts with formation of rosettes. Rat grafts are adequate animal models of human grafts and we are presently attempting to find the maximum safe cryopreservation period for fetal tissue banking. (Supported by the Grant in Aid for Special Research Project on Environmental Science from Ministry of Education, Japan.)

NISHIMURA, H., K. ARISHIMA, C. UWABE and M. ADACHI, Central Institute for Experimental Animals, Kawasaki, Kanagawa, and Department of Anatomy, Azabu University School of Veterinary

OKAJIMA, M., Department of Forensic Medicine, Tokyo Medical and Dental University, Tokyo. Methodological improvement for dermatoglyphic observations of fetal limbs. Epidermal ridges appear in the 11th week after fertilization a s undulations at the dermo-epidermal junction first o n the finger apex, and then extend over the whole volar surface by the 17th week. In the initial stage, the ridged structure does not project onto the epidermal surface which is covered with flat periderm. The traditional methods of dermatoglyphic observation, such as reconstruction of serial histological sections and exposure of the deep epidermal surface or the dermal surface by chemical abrasion, are not practical for the whole volar surface. The method proposed by the author ('75) to inspect the dermal surface stained with t o l u idine blue has enabled dermatoglyphic findings that were reported for fetuses with chromosome anomalies diagnosed by amniocentesis (Suzumori, '80; Okajima et al, '81). An improvement on the original method has been developed to reduce incomplete removal of the epidermal cells and excessive mechanical manipulation of the dermis. The new method involves increasing the time of alkaline solution treatment from one to two hours and extending it to 20 hours. With this revised method, intermediate and basal layers of the epidermis are easily removed, yielding specimens of high quality in ridge details of fetuses with soft dermal tissue undergoing initial differentiation of dermal ridges.

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SAKIYAMA, Y., I. TADA, C. T. LIAO and M . ODA, Department of Oral Anatomy, Osaka Dental University, Osaka. An experimental study on tooth morphogenesis. Morphology of mammalian tooth is determined by a definite developmental potential of the class to which the tooth belongs. Glasstone (1963), using in vitro culturing methods, found that an anterior tooth germ developed into an incisor-like tooth, while a posterior tooth germ developed into a molarlike tooth. Therefore, tooth morphogenesis was considered t o be dependent on the tooth germ, and not to be associated with the jaw bone. In contrast, Butler (1939) and Dahlberg reported that dimension and morphogenesis were affected by the jaw bone. Then, Kirino and Masuda (1959, 1967), using implantation studies of tooth germ cells, reported that tooth morphogenic factors existed not only in the jaw bone but also in the tooth germ in its initial stage of development. Studies of tooth morphogenesis using implantation techniques on 32-95 day old dogs showed the following: 1) A tooth germ undergoes development and eruption when implanted into the bony socket of another tooth class. 2 ) The morphogenesis of the implanted tooth germ retains its original shape through development and eruption. 3 ) A molar tooth germ under initial mineralization, when implanted into the bony socket of a cuspid, develops into the morphology of a cuspid. 4) Both the tooth germ and its location in the jaw bone influence tooth morphogenesis. 5) A supernumerary tooth resulting from division of a tooth germ resembles the morphology of the tooth class where the supernumerary tooth is implanted in the jaw bone.

NAKASHIMA, K., T. GOT0 and Y. FUJIKI, Department of Oral Radiology, Gifu College of Dentistry, Hozumi, Gifu, Morphogenesis of in vitro developing mouse embryos from earlysomite-stage to ear vesicle formation. The early-somite-stage ( 1 to 2 somite s stages t :ages = 1-2 S) STD-ddy mouse embryos were cultured-in They rotating tubes (38'C, 30 - 40 rev./min). were examined by light and scanning electron microscopy every 2 h for 36 h. In vivo specimens, which were equivalent to in vitro developing embryos, were also examined. The medium consisted of 80% STD-Wistar female rat I.C. serum and 20% Earle's solution, which was supplemented with streptomycin sulfate (50 pg/ml) and glucose (final concentration = 2.0 mg/ml). The gas phase was 5%0~/5%COz/90%N2 for the first 20 h, followed by a 20%0~/5%CO2/75%Nzgas mixture. Mouse embryos developed to 22-24 S during the 36 h culture period. The formation of a new somite pair required about 1.6-1.7 h. The heart began beating at 5-7 S (6-10 h) and the blood was circulating in the yolk sac at 8-9 S (14 h ) . The neural tube was fused for the first time at the cervical region at 8-early 9 S (12-14 h). The cranial neural tube was ) . The first sign of fused at 16-18 S (26-28 h optic vesicle formation was seen at 5-6 S (68 h) as a pair of indentations in the prosencephalic region. The otic vesicles were almost ) . These developmental closed at 23-24 S (36 h events closely resembled those for embryos in vivo.

SHIMIZU, T., A . TAKA0 and A. HIRAYAMA, Department of Pediatric Cardiology, the Heart Institute of Japan, Tokyo, and Department of Clinical Pathology, Tokyo Women's Medical College, Tokyo. Peculiar face, conotruncal anomalies of the heart and thymic involution without hypoparathyroidism of thymic hypoMASUDA, Y., S. SO, Y. KOJO, Y. OGURA and Y. KAMEYAMA, Department of Otorhinolaryngology, Okayama University School of Medicine, Okayama, and Research Institute of Environmental Medicine, Nagoya University, Nagoya, Aichi. Normal development of the stapes in the mouse. Fifty ddN and CF mouse fetuses were examined between day 7 and 18 of gestation. Fetal heads were serially sectioned and stained with hematoxylin and eosin. Primordia of 3 ossicles and the otic capsule appeared on day 12. The stapedial primordium was observed as a mass of mesenchymal cells lateral to the primordium of the otic capsule and attached to the medial part of the facial nerve. On day 14, the stapedial footplate was first observed as a dual structure. On day 15, the primordium became more demarcated. The possibility of the dual origin of the stapedial footplate was examined in normal development of the human stapes. (Supported in part by a Grant in Aid for Scientific Research from the Ministry of Education of Japan.)

plasia.
We reported, in 1980, the unusual facial appearance specifically related to conotruncal anomalies of the heart, and named it conotruncal anomaly face (CTAF). Seven females and nine males, aged 5 months to 11 years-old with these facial features also had 15 incidences of tetralogy of Fallot (6115 right aortic arch) and 1 transposition of the great arteries. Studies also included phytohemagglutinin (PHA) skin tests and thymus biopsies. Results: 1) There were no complicated pregnancies. 2 ) All subjects had CTAF and conotruncal anomalies of the heart. 3) Serum calcium levels were within normal limit except in one instance. 4) Poor responses to PHA skin tests were noticed in 11 patients. 5) Distinction between thymus cortex and medulla was blurred and cystic Hassall's corpuscles increased in number. Discussion: The poor responses to thymusdependent PHA skin tests, peculiar faces characterized by ocular hypertelorism, flat nasal bridges, small mouth and deformed ears and conotruncal anomalies of the heart led u s to 1 1 study the association between CTAF and the 1

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N pharyngeal pouch syndrome. However, the CTAF is somewhat different from the I=- IV pharyngeal pouch syndrome. The CTAF shows good life expectancy and no signs of hypoparathyroidism. Further studies are needed to clarify the relationship between the CTAF and the m N pharyngeal pouch syndrome.

and outer adrenal cortical cysts, other anomalies were observed, including prematurity, chromosomal abnormalities, and bacterial and viral infections. (Supported in part by the Japanese Ministry of Health and Welfare.)

SUZUKI, S., T. YAMAMURO and S. MATSUURA, Department of Orthopedic Surgery, and Department of Obstetrics and Gynecology, Faculty of Medicine, Kyoto University, Kyoto. A causative factor o f congenital dislocation of the hip and torticollis. Congenital dislocation of the hip (C.D.H.) and torticollis are the most common diseases in the field of pediatric orthopedics. We analyse the etiologic factors of these two conditions from an obstetrical point of view. Materials and Methods: The clinical material consisted of 1187 infants born from July 1978 to April 1980. The incidences of C.D.H. and of torticollis, fetal X-ray films, and ultrasonography of fetuses were studied according to the type of the delivery. And the heredity of the various types of delivery was investigated. Results: The incidences of C.D.H. and torticollis were significantly higher in the neonates born bv single breech presentation than those hv other types of delivery. As for the fetal position in the uterus, extended cervical spine and extended knees were closely related to the single breech presentation. Conclusion: The development of C.D.H. a s well as torticollis is greatly influenced by the intra-uterine posture of the fetus, their incidence being markedly higher in the neonates delivered by single breech presentation. The extended posture of the knees and cervical spine seems to be the most important prenatal causative factor of C.D.H. and torticollis.

YASUDA, Y. and T. TANIMURA, Department of Anatomy, Kinki University School of Medicine, Sayama-cho, Osaka. Effect of ethinyl estradiol on the differentiation of mouse testis exposed in utero. Pregnant Jc1:ICR mice were either given oral doses of 0.02 or 0 . 2 mg/kg body weightlday ethinyl estradiol (EE) in olive oil or olive oil alone (controls) on day 11 through day 17 of gestation. One testis from each animal in the treated or control group was sectioned in five regions each separated by 4 0 u m . The following determinations were made using the Texture Analyse System (T.A.S., Leitz). 1. The ratio between the area of the seminiferous tubuli and the stroma was calculated. 2. The ratio between the number of germ cells and Sertoli cells in a cross-sectioned tubulus was 0 0 Leydig determined. 3 . The mean size of the 1 cells selected at random in the stroma was recorded. The ratio between the tubular area and that of the stroma decreased in the EE exposed testes. The number of germ cells per tubulus was significantly increased in the drug treated group and showed a dose-response relation to EE. The ratio between the number of germ cells and Sertoli cells per tubulus was significantly higher in the treated group than in controls. The mean size of Leydig cells was smaller in the treated group than in controls. These findings suggest that prenatal exposure to EE affects tubular formation, stimulates proliferation o f germ cells and affects the function of the Leydig cells and the antimiillerain substances.

OBARA, A., T. ITOH, M. KITAICHI, H. ASAMOTO and M. FURUTA. Deoartment of Patholoev. National

= o n in the thymic cortex o f neonates and early infants is a phenomenon caused by prominent swelling of reticuloepithelial cells, suggesting early changes of thymic involution (presented at the 71st Annual Congress of the Japanese Pathological Society, 1982). A survey of 400 autopsy cases of neonates and young infants was performed to identify outer adrenal cortical cysts (Congenital Anomaly, Vol. 16, No. 2, 1976) combined with the starry sky phonomenon. We found more than 50 cases with the starry sky phenomenon of the thymic cortex; more than 80 cases with outer adrenal cortical cysts; and more than 30 cases with both abnormalities. Among the cases with both starry sky phenomenon

MATSUMOTO, N., S. KATAYAMA and H. KUBO, Department of Public Health, Faculty of Medicine, University of Tokyo, Tokyo, and First Department of Obstetrics and Gynecology, Toho University, School of Medicine, Tokyo. Toxic effects of methylmercuric chloride (MMC) and mercuric chloride (MC) on the development of early mouse embryos in vitro. Late blastocysts were treated with mercury compounds at various concentrations for 2 4 hrs, then cultured for another 120 hrs with mercury free medium. Blastocyst cell number was counted after a 24 hr culture with mercury free medium. Ultrastructual changes of the surface of the zona pellucida were observed using scanning electron microscopy. The development of the inner cell mass (ICM) to two layers was inhibited with lxIO- M of MMC, while trophoblast outgrowth and ICM development were not affected at the same

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concentration. At ZxlO- M of MC, trophoblast outgrowth and the development of ICM to two layers were inhibited. Blastocyst cell numbers treated with MMC at IxlO-M to 2x10d6 M were about 30% that of control, but, at 2x10 -6 M of MC, development of blastocysts was not affected. Cell numbers were down to 30% at 2x10 -5 M of MC. Blastocysts started to collapse after a 5.5 M ~of MMC and slight hr culture with ~ x I O changes in fine net-like structure of zona pellucida came out. After 24 hrs in culture, the blastocoel collapsed and the zona lost its net-like structure. These findings were observed in the zona pellucida of blastocysts M~ of MC. cultured for 24 hrs with 2 ~ 1 0 (Supported by a Grant-in-Aid for Environmental Science, the Ministry of Education, Science and Culture, Japan.)

IIJIMA, S., A. SPINDLE and R. A. PEDERSEN, Department of Helath Sciences, Medical University of Yamanashi, Tamaho, Yamanashi, and Laboratory of Radiobiology and Environmental Health, University of California, SanFrancisco. Developmental and cytogenetic effects of potassium dichromate on mouse embryos in vitro. Mouse blastocysts and egg cylinders were treated in vitro with potassium dichromate to 2 x M) and the (K2Cr207, 5 x effects on development and induction of sister chromatid exchanges (SCEs) were determined. When blastocysts were treated for 6 days M, the number of embryos with with 1 x growing inner cell masses (ICMs) and 2-layer ICMs were significantly reduced. The number of embryos forming trophoblast outgrowths was not reduced, but outgrowths formed in the presence of 2 x M were small and eventually degenerated. Hatching and attachment were not affected at any concentration, The frequency of SCEs was significantly increased at 2 1 x 1 W 6 M . When egg cylinders were treated with KzCrz07 for 24 hours, the number of embryos with blood islands, fusion of the allantois with the chorion, and beating heart was significantly reduced at all concentrations tested. The frequency of SCEs was increased in the cells of embryonic tissues of egg cylinders cultured in 21 x M. Extraembryonic cells showed a higher frequency of SCEs than embryonic tissues at all consentration of KzCr207. The correlation between developmental effects and SCE frequency suggest that cytoFenetic damage may be largely responsible for the detrimental effects of dichromate on mouse embryos. The high sensitivity of embryonic development to all the endpoints indicates the usefulness of embryo culture in teratogenicity and toxicity testing. (Supported by the U.S. Department of Energy)

Previous teratological observations with nicotine in chick embryos demonstrated various malformations, notably skeletal abnormalities, hydrops and eventration. The finding of eventration of the abdominal viscera prompted further investigation of the underlying teratogenetic mechanism. Fertilized chicken eggs were injected intra-albumenally with 3 mg of nicotjne per egg and incubated at 38C. Developing embryos were examined for external deformities, especially for morphologic abnormalities of the thoraco-abdominal walls, at early middle and late stages of development in ovo. After 4 days of incubation with nicotine, extraembryonic blood vessels showed evidence of stagnant blood flow as compared to controls. Embryos with abdominal wall dysgenesis were found at 7 days of incubation. The abdominal skin and muscle layers were so thin that the abdominal viscera were barely covered with the amnion o r peritoneum at mid-incubation period. During the late incubation period thoracoceloschisis, eventration, and malformation of the spinal skeleton were prominent. This observation suggests that nicotine might exert teratogenic actions upon the somites o r somatopleure in a very early stage of embryonic development to jnhibit their differentiation and development, leading to the occurrence of celosomia in embryos.

TERAKI, Y., T. CHIBA, J. TANAKA and K. NAGUMO, Department of Anatomy, St. Marianna University School of Medicine, Kawasaki, Kanagawa. Nicotine-induced celosomia in chick embryos.

YOSHIDA, K., K. MATAYOSHI, T. KIYOKAWA and H. SOMA, Department of Obstetrics and Gynecology, Tokyo Medical College, Tokyo. The correlation of developmental variability of twin fetuses and placental vascular communications. In twin pregnancies, developmental differences between fetuses are commonly observed, and is conspicuous in monozygotic twins. Different growth rates of twins can be ascribed to zygosity, the structure of twin placentas and communications of fetal vessels in monochorial twin placentas. The purpose of the present study was to examine the presence of placental vascular communications relating to twin transfusion syndrome in order to clarify some causes of developmental discrepancies between twins. In 105 cases associated with monochorial twin pregnancies, vascular placental communications occurred in 78 cases. Sixteen cases were diagnosed to be twin transfusion syndrome, and a marked discrepancy of fetal size of twins was noted. In 10 of 16 pairs, arterial-toarterial and Venus-to-Venus communications of the placenta were demonstrated by a perfusion method. In 5 other pairs, artery-to-vein shunts with o r without artery-to-artery anastomosis were recognized. In only one case, a single vein-to-vein communication was found. Unequal blood distribution between paired twins tends to occur in cases which have a vascular communication of vein-to-vein OK artery-to-vein shunts as well as artery-to-artery anastomosis in the placenta. It is possible that developmental differences and fetal anomalies occur as a result of abnormal vascular communications.

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MOMOTANI, N. and K. ITO, Ito Thyroid Clinic and Hospital, Tokyo. Maternal thyroid function and congenital malformation in the offspring. FOUK hundred and forty six (446) neonates from mothers with Graves' disease of Hashimoto's thyroiditis were examined for malformations of external organs to determine whether maternal dysfunction during the first trimester could cause congenital malformations. Neither infants born to mothers who received radiation or drugs nor multiple births were included. None of the mothers were diabetic. Thyroid function was determined mainly as Free TI, Index and partly as the total Tr and T3 concentration, due allowance being made for the influence of pregnancy on the normal range of values. Statistical significance was assessed by Fisher's exact test. O f 50 infants from hyperthyroid mothers, 3 (6.0%) revealed malformations, while 4 ( 1 . 1 % ) of 377 infants from euthyroid mothers revealed malformations. The types of malformations were not specific. The incidence was significantly higher in the hyperthyroid group than that in the euthyroid group (p<O.O5). None of 19 infants from hypothyroid mothers was found to have malformations. There were no differences both in the mean values and distributions of maternal ages among the three groups. Although the correlation between the frequencies of malformations and the levels of hyperthyroidism could not be investigated because of insufficient numbers, the above finding suggests that maternal thyrotoxicosis causes congenital malformations. The numbers were too small to examine the causal relationship between maternal hypothyroidism and congenital malformations in the offspring.

and visceral malformations that were attributed to the mother's malnutrition. Neonates were breast-fed by mother rats for about one month. The group receiving a diet including 1 0 % hydrogen peroxide showed abnormal growth. The results of the present experiments show that hydrogen peroxide, given to pregnant rats, can cause growth retardation of neonates.

MORIYAMA. I., M. FUJITA, K. HIRAOKA, M. ICHIJO and S. KANOH, Department of Obstetrics and Gynecology, Nara Medical University, Kashihara, Nara, and Department of Pharmacology and Microbiology, National Institute of Hygienic Sciences, Osaka Branch, Osaka. Effects of the food additive hydrogen peroxide on fetal development. HydroKen peroxide is widely used, especially as a food additive for bactericidal preservation of food and as a bleaching agent. Experiments with rats were performed to study the biological effects of hydrogen peroxide on pregnancy. Tests for acute toxicity were performed in four groups with hydrogen peroxide orally administered at l o % , 2%, 0.1%, 0.02%. There was no indication of acute toxicity except for a significant decrease in body weight with 10% hydrogen peroxide. Pregnant rats received, for one week at the critical period, food containing l o % , 2 % , 0.1%, 0.02% of hydrogen peroxide respectively, and examined at the 20th day of gestation. External fetal features were observed, and visceral and osseous conditions were examined with Wilson's method and Dawson's method, respectively. 10% hydrogen peroxide in the diet caused lowered body weight and some instances of bone

HASE, Y., T. TSURUHARA, T. OUW, S. YAMAGUCHI, K. UEDA and Y. KOBAYASHI, Department of Pediatrics, Department of Laboratory Science, Department of Cardiology, and Department of Pathology Children's Medical Center of Osaka City, Osaka. Maternal PKU treated from early pregnancy. We report two offspring, one fetus and one newborn, of a phenylketonuric mother of normal intelligence, who was first diagnosed when she was 30 years old. At that time she was in the 7th month of her third pregnancy and had two previous miscarriages. Her pregnancy was artificially interrupted on medical indication and the fetus was autopsied. The fetus had no intrauterine growth retardation and no congenital malformations, but his brain weighed 155 g, which was at the lower limit of brain weight. On a normal diet, her serum phenylalanine (Phe) level was 24.5 mg/dl, her serum tyrosine was 0.13 mg/dl and her urine ferric chloride test was positive. In the fourth pregnancy a Phe-restricted diet was instituted at 8 weeks of gestation, however spontaneous abortion ensued about 4 wks later. In the fifth pregnancy, dietary treatment was started at 6 weeks of gestation, using Phe-free Lophemilk and amino acid mixed powder. The diet provided 80 g protein, 2200 kilocalories and 750 mg phenylalanine per day. Serum Phe remained below 10 mg/dl throughout pregnancy. Despite this early treatment, her fullterm male baby had intrauterine growth retardation, multiple cardiac malformations (hypoplastic left heart syndrome), 1 1 pairs of ribs and a 10th thoracic hemivertebra. He died at 10 days after birth and was autopsied. His brain weighed 345 g which was normal in size, but the cerebellum was small. Microscopic examination showed a maturation arrest of nerve cells. An electron-microscopic examination demonstrated n o apparent hypomyelination of nerve fibers.

KIDA, M., M. UEHARA and K. YOSHIMURA, Department of Pediatrics. Facultv of Medicine. Teikvo University, Tokyo. Analysis of clinical pregnancy records of mothers with malformed children. Clinical pregnancy records of 62 cases taking medication in the first trimester of pregnancy were analyzed. The types and numbers of malformations were: 20 with peripheral hypoplasia of limbs, 12 with constriction ring syndrome, 7 with Poland syndrome, 4 with radial

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defects, 3 with ulnar defects, 3 with split hand and foot, 3 with symbrachydactyly, 3 with arthrogryposis multiplex congenita, and 2 with ear anomalies. 109 different medicines from 23 drug categories were prescribed during treatment. The number of prescriptions were counted for each type of malformation. In total, 20 cases of peripheral hypoplasia of limb received 2 categories of psychotropic drugs 5 times, 1 category of antidote 2 times, 1 category of non-sex hormonal agent 2 times, 3 categories of cardiovascular drugs 5 times, and 13 categories of sex hormonal agents 15 times. 12 cases of constriction ring syndrome recei.ved 1 category of antidote 3 times, 3 categories of cardiovascular drugs 5 times, and 1 1 categories of vitamins 15 times. (This study was supproted by the Association of Parents of Children with Limb Malformations.)

logical diagnosis by ultrasonography and functional diagnosis by FHR monitoring and fetal ECG were performed. When abnormalities were found, additional tests were done using chemical, endocrionological and genetic methods. We have diagnosed in utero intestinal obstructions, non-immunologic fetal hydrops, hydrothorax and hydrocephalia with meningocele using ultrasonography, and functionally fetal sinus tachycardia using FHR monitoring and fetal ECG. These cases, except for hydrocephalia, were treated soon after birth. However, all congenital abnormalities could not be diagnosed in utero with these procedures. Further investigations are required.

TAKAUCHI, N., T. HONDA, Y. INOMATA, and S . TAKEUCHI, Department of Gynecology, Saiseikai Hospital, Niigata, and Department of Gynecology, Faculty of Medicine, Niigata University, Niigata. Clinical analysis of consanguineous marriages in genetic counselling. Until the end of 1981, 732 individuals visited our genetic counselling clinic in the gynecologic outpatient service of the University Hospital. Thirty four individuals (4.6%) were counseled for consanguineous marriages. The genetic counselling section in the Public Health Center of Niigata City counseled a total of 160 individuals of which 22 (13.77:) were for consanguineous marriages. Clinical analysis of these 56 cases was undertaken. The intensity of the inbreeding coefficiency varied from 7/64 to 11128, most of them giving a value over 1/16. Some other clinical features of the analysis are t o be presented.

IWAMOTO, K., M. OHTA, N. MIYAMOTO, K. TAKEUCHI, T. TSUZAKI and K. MAEDA, Department of Obstetrics and Gynecology, Tottori University School

KITAGAWA, T., M. SAITO, M. ARIMA, Y. ETO, T. ORII, T. KUSUNOKI, Y. SUZUKI, K. TADA, M. YABUUCHI and S. TARUI, Departments of Pediatrics, Nihon University, National Center for Nervous, Mental and Muscular Disorders, Jikei University, Tokyo, Gifu University, Gifu, Kyoto Prefectural University of Medicine, Kyoto, Tokyo University, Tokyo, Tohoku University, Miyagi, Osaka University, Osaka, and Department of Internal Medicine, Osaka University, Osaka. Prenatal diagnosis in Japan of inborn errors of metabolism. 127 cases (23 disorders) have been examined in utero for inborn errors of metabolism in the 0 years. 93 fetuses (73.2%) were diaglast 1 nosed as normal and 34 fetuses (26.8%) were diagnosed as having a metabolic disease. In our experience, Tay-Sachs disease, Gaucher disease, GMi-gangliosidosis and I-cell disease were the most common disorders, and accounted for about 57% of all cases. In Japan, Prenatal diagnosis of inborn errors of metabolism have been performed using enzyme activity of cultured cells from the amniotic fluid. In six of 133 amniocenteses c e l l s in the amniotic fluid would not grow in culture. Three of the 127 prenatal diagnoses f o r inborn errors of metabolism were incorrect because cultivated cell counts were not enough for the diagnosis. Since 1976 there has been an increase in the number of pregnant women at risk for inborn errors of metabolism who request prenatal diagnoises, because of the reliability and safty of the diagnostic method.

abnormalities are desirable for the care of pregnancy, decision of the method of delivery, early detection of problems and treatment of the newborn infant. Recently, antenatal diagnosis of congenital abnormalities by ultrasonography, FHR monitoring and fetal ECG has been tried on many cases. We planned a screening system to diagnose fetal congenital abnormalities in utero with those procedures. In the first trimester, gestational age was estimated by ultrasonic measurement of fetal bones. In the 2nd and 3rd trimesters, evaluation of fetal growth, morpho-

SUZUMORI, K., T. KOISHI, K. IMAIZUMI and Y. YAGAMI, Department of Obstetrics and Gynecology, Medical School, Nagoya City University, Nagoya. Prenatal diagnosis of the balanced translocation carries: Results of analyses of fetal cells and segregation analyses. The highest risk of abnormal finding is observed among those families in which one of the parents is a balanced translocation carrier. During the past 12 years, there were 41 prenatal diagnoses of parental chromosome translocation among 550 pregnancies, a rate of 7.6%;

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31 observations with a parental Robertsonian translocation, D/21, 13/14, 13/15, 21/22 and 10 observations with a parental reciporcal translocation, t(5;16), t ( 8 ; X ) , t(3;11), t(4;15) and t (3;5). Among 34 prenatal diagnoses at risk with a female carrier, 11 unbalanced fetuses, showing a 26.8% incidence, were diagnosed. By contrast, no unbalanced offspring was detected in 7 prenatal diagnoses with a male carrier. Nine of the 11 unbalanced fetuses were diagnosed in the mothers with Robertsonian translocation, D/21, the other 2 were detected in the mothers with reciprocal translocation, t(5;16), t(3;ll). Our study suggests that behavior in gametic segregation varies considerably according to the sex of the carrier and the type of translocation. Continuous data accumulation is needed for more precise segregation analysis of parents to identify a translocation carrier for genetic counseling.

SHIRAI, Y., and N. FUJIKI, Department of Social Welfare, Institute for Developmental Research, Aichi Prefectural Colony, Kasugai, Aichi, and Department of Internal Medicine, Fukui Medical School, Matsuoka, Fukui. Japanese women's attitudes toward prenatal diagnosis and selective abortion. Prenatal diagnosis has made an important contribution to genetic medicine and has resulted in effective preventive measures of congenital malformations. Recently, the Ministry of Health and Welfare set about providing a guideline on genetic counseling. However, the opinions of the lay public are largely unknown. The present study provides an analysis of women's attitudes on prenatal diagnosis and selective abortion of high risk fetuses. The study included 407 married women less than of 39 years old residing in Nagoya City or Kasugai The results were as follows: 1) Attitudes toward prenatal diagnosis: A breif explanation of the diagnostic techniques was followed by a description of an actual case in which a young woman, pregnant with her second child, underwent prenatal diagnosis after her first child was born with Down's syndrome. Regardless of age and education, 89% of respondents approved of prenatal diagnosis. 2) Attitudes toward selective abortion: Eightyeight per cent of respondents approved of abortion if a fetus had a serious congenital defect. At the same time, a significant positive correlation was shown between educational level and attitude toward selective abortion. 3) Right to life of a defectlve fetus: Only 28% of the respondents were in favor of the right to life of a high risk fetus in the first trimester. It was observed that education has a strong influence, with better educated respondents most likely to disapprove of the right to life of a defective fetus.

The Japan Association for Maternal Welfare investigated congenital malformations reported in 130 hospitals since January 1972. The number of cooperating hospitals increased to 236 since January 1979. Statistical analysis of congenital malformations from January 1972 to December, 1981 has been reported. The total number of births during that time was1,009,200. 7,939 cases of congenital malformations were reported. The incidence rate was 0.787%. Of these congenital malformations, the incidence, in order from most to least frequent, was as follows: cleft palate, cleft lip, anencephaly, polydactylia (finger), extremities, others, syndactylia (toe), ears, others, Down syndrome, polydactylia (toe), anal atresia.

KONDO, K., K. KATO, T. AKITA, and T. KUROSU, Department of Neurology, Tokyo Metropolitan Institute for Neurosciences, Fuchu, Tokyo and National Institute for Minamata Disease, Minamata, Kumamoto. Monitoring of congenital malformations in the Metropolitan hospitals in Tokyo: Further observations. As reported at the annual meeting of the Japanese Teratology Society in 1979, congenital malformations recognized during the first postnatal week in all live and still births have been exhaustively recorded in 11 Metropolitan hospitals since April 1, 1978. By December 31, 1981, a total of 3,783 malformations was recorded among 44,534 infants including 647 stillborns. Malformed infants totaled 3,218 or 7.2% of all births, with the average number of 1.18 malformations per capita. 419 had multiple malformations includ5 monomeriing 39 chromosome anomalies and 1 cally inherited syndromes. Major malformations comprised about 35% of all recorded malformations, and were observed in about 2.5% of all births, the proportion being comparable with those in other populations. The annual incidence rates o f 12 types of malformations recommended by the International Clearinghouse for Birth Defects Monitoring Systems were stationary during 4 years, at a 1% alarm level. Studies are underway to evaluate detailed frequency patterns including areal and seasonal incidences, familial clustering, etc. A casecontrol study is being considered to evaluate risk factors in selected malformations.

SUMIYOSHI, Y., Y. MORIYAMA, M. GOMIBUCHI, G. NOZUE, S. MINAGAWA, A. KUROSHIMA, T. KIYOKAWA, and Y. NOTAKE. Japan Association for Maternal Walfare, Tokyo. The statistical

SATO, K., F. KAYAMA, Y. MORITA, M. MIZUNO and S. SAKAMOTO, Department of Obstetrics and Gynecology, School of Medicine, University of Tokyo, Tokyo. Questionnaire survey of clients

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before genetic counselling. II. Understanding and anxiety about genetics and antenatal diagnosis. Understanding and anxiety of clients about genetics and antenatal diagnosis were studied by means of questionnaires before genetic counselling. (1) About 6 percent of the clients kept their visits to our clinic secret from their spouses. (2) The recurrence risk evaluated by the clients was much higher than that estimated by counsellors in about half of the clients. (3) About 12 percent of the clients who wanted to have amniocentesis had decided not to bear another child if amniocentesis was not available. (4) Eight percent of the clients did not permit us to follow up.

nancies subjected to chromosome analysis, one hundred and sixty had the procedure because of earlier births with regular 21-trisomy syndrome. Two fetuses were detected as abnormal. One affected-fetus and three carrier-fetuses were found among 8 pregnancies subjected to amniocentesis because of chromosome translocations in the parents. Twenty five pregnancies were examined because of advanced maternal age (over 4 0 ) ; 2 fetuses were abnormal. In 13 pregnancies, amniocentesis was performed because of X-linked recessive disorders. In twenty four pregnancies, amniocentesis was performed because earlier births had shown genetic metabolic disorders; 8 affected-fetuses and 4 carrier-fetuses were detected.

TSUCHIYA, S., S. YAMADA, A. AKIMARU, K. FUJIKI, K. KATO and A . NAKAJIMA, Department of Ophthalmology, School of Medicine, Juntendo University, Tokyo. Estimation of the prevalence of congenital cataract among Japanese children. Congenital cataract is one of the most important eye djseases causing blindness. According to a nation-wide research of causes of blindness among children in schools for the blind, 1186 (14.0%) of the 8464 children examined had congenital cataract in 1975, and 1062 (13.6%) among 7799, in 1980. All cases showed bilateral involvement with visual acuity under 20/200. Therefore, it was difficult to obtain the number o f unilateral o r bilateral children whose visual acuity was better than 20/200. In the present study, we estimated the prevalence of congenital cataract in the Japanese population by the following methods. 1. Estimation from the number of conpenital cataract among children in 1980, combined with information on the ratio of bilateral and unilateral cases as well as the distribution of visual acuities (in case a patient had operation, the post-operative visual acuity was taken) obtained from cases in Juntendo University. 2. Analysis of the probability of ascertaining the number of proband in the research of schools for the blind in 1959 and 1964. 3. Estimation of consanguineous marriages among normal parents and the segregation analysis of the families in 1959 and 1964. These results were compared with the ophthalmologic surveys on primary and secondary school children in several areas. The prevalence of congenital cataract in Japanese children was (2-5) x ~ O - ~ .

SHIOMI, B., G. NOZUE, M. KITAMURA, S . NESHIME, M. ASHIZAWA and M. KIMURA, Japan Red Cross Hospital and Institute of Public Health, Tokyo. A clustering of cases of dysplastic genital anomalies. A monitoring system has been set up in 5 Red Cross Hospitals in the Tokyo area since 1976, and 5 cases of pseudo-hermaphroditism/dysplasia of external genitalia were found. 4 cases were clustered between May 1976 and February 1977; all were stillborns with birthweights less than 2000 g. In addition, these cases also had multiple anomalies of the ear, lip/palate, limb, anus and diaphragm. The cases also were suspected of having the VACTEL syndrome. Intake of hormonal drugs during early gestational period was reported in one case. Another case was registered as an early neonatal death with diaphragm hernia in January 1981. We noticed the unusually high incidence of hermaphroditism (6 cases) and limb/musculoskeletal anomalies (15 cases) observed out of 4920 total deliveries in 1959 from the medical records of the Japan Red Cross Lying-in Hospital, Tokyo. During the above-mentioned period, exogeneous hormons seemed to have been widely used prenatally. Further analysis will be done on the new clustering of anomalies that occurred around 1976.

KAYAMA, F., Y. MORITA, K. SATO, T. JIMBO, M. MIZUNO and S . SAKAMOTO, Department of Obstetrics and Gynecology, School of Medicine, University of Tokyo, Tokyo. Genetic amniocentesis at the University of Tokyo Hospital. Genetic amniocentesis was carried out for 319 pregnancies between 1973 and 1981 at the Department of Obstetrics and Gynecology, University of Tokyo Hospital. Out of 235 preg-

NARUSE, Y., S. KAGAMIMORI. M. WATANABE, T. SHOJI and 0 . MORITA, Department of Community Medicine, Faculty of Medicine, Toyama Medical and Pharmaceutical University, Toyama, and Toyama Institute of Health, Toyama. Factors in the occurrence o f congenital anomaliesA prospective study was carried out in Toyama prefecture to study environmental factors associated with congenital anomalies. 1,046 pregnant women were examined for clinical findings during the course of pregnancy and delivery, and for serum antibodies against rubella virus (R.V.) and Toxoplasma (T.P.). There were 4 abortions and 10 perinatal deaths, but no specific factor relating these cases was found.

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142 babies (13.7%) had anomalies of the following systems; musculoskeletal system 7.9%, genital organs 1.5%, circulatory system 1.1% and central nervous system 0.8%. There was a higher incidence of anomalies of the extremities in females than in males, and in breech presentation than in vertex presentation. The incidence of anomalies was also higher among infants whose mothers were negative for the R.V. antibody and positive for the T.P. antibody during mid pregnancy. However, this difference was not observed when the infants were stratified according to the antibodies in serum of cord blood and in maternal serum taken at delivery. Cesarean sections were seen as a risk factor for all anomalies.

asymmetrical face, hip joint limitation and radial nerve neuraplaxia were most often seen. The incidence of CPD is higher than the incidence of congenital malformations. CPD can be corrected spontaneously during the infantile period.

OKAMOTO, N. and Y. SATOW, Department of Geneticopathology, Research Institute for Nuclear Medicine and Biology, Hiroshima University, Hiroshima. New standards for the classification of autopsy population of human fetuses and newhorns. . . - .- - - ..-. The autopsy population of human fetuses and newborns, amounting to about 10,000 cases, was examined to identify basic fetopathological samples and to devise new classification standards. In our laboratory, artificially aborted fetuses with no abnormal findings were selected as control fetuses; fetuses with some anomalies, seen at autopsy, were labeled as anomalous fetuses; fetuses with abnormal pathological findings were identified as abnormal fetuses. We followed the classification of anomalies described by Schwalbe and Hayashi where anomalies were divided into two groups; simple and double monsters. The double monsters were subdivided into conjoined and non-conjoined twins (monozygotic twins). Artificially or spontaneously aborted twins were separated from the autopsy population. Recent changes of environmental conditions such as improved nourishment of mothers and a decrease of mothers with infectious diseases, were considered. New control standards were determined for anomalies and abnormal findings by inferring age by height and weight of the fetuses and newborns.

KINOSHITA, S., K. SHIRAI and T. MATSUO, Department of Pediatrics, Nagasaki Municipal Hospital and Department of First Pathology, Faculty of Medicine, Nagasaki University, Nagasaki. Metaphyseal chondrodysplasia and spondyrometaphyseal dysplasia. We report here two cases of osteochondrodysplasia which each have metaphyseal dysplasia. Case 1: A male, 5 months old, was examined for his bowed legs. His skeletal radiogram demonstrated widened and cupshaped metaphysis of the long bones and he was diagnosed as having rickets. He receiveda-D3 therapy for two years with no success. From a detailed examination, we changed our diagnosis to Schmid type metaphyseal chondrodysplasia. A bone biopsy of the iliac crest was performed and showed increased chondrncyte nests, and blood vessels in the matrix of the epiphyseal plate. Electron microscopically, the chondrocytes of the growth plate demonstrated numerous rough endoplasmic reticulum. A 1 2/3-year-old male was seen Case 2: because of his wadding gait. His skeletal radiogram demonstrated platyspondylia of the spine and an irregular metaphysia of the long bones. He had no symptoms of ophthalmology, and analysis of urinary mucopolysaccharides was normal. He was diagnosed as having Kozlowski type spondylometaphyseal dysplasia.

MINE, M., S . MOOCHIZUKI, H. TAKASHIMA and K. BABA, Department of Pediatrics, Faculty of Medicine, Nihon University, Tokyo. Clinical observations of congenital postural deformities. Congenital postural deformities (CPD) are caused by intrauterine moulding. Two typical cases of a congenital moulding depression of the skull were observed in our study. They were from normal full term pregnancies. In both cases, a large depression, 3 . 5 x 4 x 4 cm, was found on the parieto-temporal region of the skull. Within three months the depression spontaneously disappeared. The incidence of CPD in full-term newborns Talipes equinovarus, calwas 5.9% (15/202). caneovalgus, overlapped fingers, midline cleft,

TANIOKA K., E . TOMOYOSHI, and T. ADACHI, Department of Pediatrics, the Japan Baptist Hosuital. and Deuartment of Patholoev. Kvoto .~ University, Kyoto. A case of complete absence of the right lower extremity. There are many types of defective anomalies of the four extremities, but complete absence of a lower extremity is very rare. Case: The second of identical female twjns. Gestational age 36 weeks 6 days. Birth weight 1600 g. Her mother was a 2 3 year-old primipara. Pregnancy was normal. The right lower extremity and right pelvic bone were completely absent. There was a large skin defect of the abdominal wall. The right lower quadrant of the abdomen was covered with a thin membrane through which the lfver and intestines could be seen. The lower part of the membrane was ruptured and the intestines and spleen were seen directly. Other anomalies were: Severe scoliosis, short sternum, bilateral simian creases, lumbar meningomyelocele and a defect of the right 12th rib. The umbilical cord was very short, about 20 cm with a single umbilical artery.
L_
~

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She died two hours after delivery. The first twin had no visible anomalies and is healthy at age 9 mo. No etiological factors could be discovered.

. . K. IZUMI, K. OKUMURA, T. SATOMI, ISHII, M K. KURIHARA and E. MARUMO, Department of Plastic and Reconstructive Surgery, Jikei University School of Medicine, Tokyo. A case report of a third leg. A very rare case of a congenital third leg from the gluteal region is described. The patient was a seven month old female. The third leg was recognised as an extra limb at birth and exhibited marked hypoplasia in comparison to the normal legs. She had bilateral fibular hypoplasia, defects of the 4th toe, clump feet and other malformations. Her parents had no history of drug usage. Pregnancy was uneventful and the infant was born after a 10 month gestation period. An operation was performed to excise the third leg and plastic surgery was performed on the toes of the remaining feet. Continuity of the soft tissue of the third leg but not the bones OK joints was seen. Reports of supernumerary legs are few and many synonyms exist for the condition, such as three legs, third leg, duplication, dipodia, pygomelus and dipygus. In the literature, cases like this are extremely rare.

AZLIMA, A., S . HOSOI, K. WATANABE, T. KOJIMA and T. TSURUTA, Department of Orthopaedic Surgery, and Department of Pediatrics, Yamada Red Cross Hospital, Misono, Mie, and Department of Orthopaedic Surgery, Mie University School of Medicine, Tsu, Mie. A case of thanatophoric dysplasia. A female infant was delivered at 42 weeks by cesarean section. The mother was a 24 year old multigravida. Her first pregnancy was hydatidiform mole two years ago. The infant's father was 3 1 years old. There was no family history of consanguinity or dwarfism, and no history of exposure to potential teratogenic agents. The newborn showed characteristic clinical and radiologic manifestations. The birth weight was 3310 g and the body length was 38 cm. The head was relatively large with a saddle nose. The extremities were extremely short with numerous skin folds. The bones of the extremities were very short and curved. The femur showed bulky epiphyses and bowing giving it a "telephone receiver" appearance. The vertebral column showed flat disc-like vertebral bodies and wide inteKVeKtebKa1 discs. The infant died four days after birth as a result of respiratory insufficiency.

MATSUZAKI, Y., I. SHIRAMIZU, K. KANEKO, K. KATO N. KATAGIRI and H. KITAMURA, Department of Obstetrics and Gynecology, and Department of Pediatrics, Yokohama Minami Kyosai Hospital, and Department of Obstetrics and Gynecology, and Department of Pathology, Yokohama City University School of Medicine, Yokohama.

dwarfism with short limbs in 1967 and utilized the Greek term thanatophoric to emphasize that such patients usually die shortly after birth. Although there are a number of cases of thanatophoric dwarfs, very few cases have been reported in siblings. A 25-year-old Japanese woman, para 2-1-1-0, recently delivered a second thanatophoric dwarf. The period of gestation was 28 weeks. The newborn infant had the features of a thanatophoric dwarf and died 2 hours after delivery. There was no family history of dwarfs, and both the husband and wife denied consanguineous marriage. The first dwarf had been delivered 4 years previously. The pregnancy at that time was normal and she delivered a 2390 g thanatophoric male with complications. He died 27 minutes after delivery.

EGUCHI, M., S. MASUMI, S. NOMURA, K. OGATA, E. SHIOTA, K. ARAI and G. TOKUHISA, Department of Orthopaedic Surgery, Faculty of Medicine, Kyushu University, Fukuoka, and Department of Orthopaedic Surgery, Oita Medical School, Oita. Spondyloepiphyseal dysplasia with osteoporosis: A new syndrome. Spondyloepiphyseal dysplasia is a heterogenous disease with no satisfactory classification. The features of a condition regarded as a clinical entity are described. The following description of spondyloepiphyseal dysplasia with osteoporosis involves the study of 7 patients (5 males, 2 females). Major findings: 1 . Short stature with normal body proportions. 2. Flat face, occasionally with wide-set eyes and depressed nasal bridge. 3. Knock knees OK bowed legs, occasionally with dislocation of the patella. 4. Shortening of one or more fingers and toes due to short phalanges; occasionally with short . Flat feet. 6 . Ligamentous metatarsals. 5 laxity. Major radiographic features: 1. Generalized osteopenia with involvement of the vertebral bodies, epi-metaphyses of long bones, carpal and tarsal bones. 2 . Generalized flattening of vertebral bodies with irregular endplates. 3 . Irregularity and flattening of the epiphyses of the tubular bones, predominantly at the hips and knees. 4 . Cortical thinning. 5. Shortening of the distal phalanges of the thumb and big toe. The data suggest that this skeletal dysplasia is a heritable autosomal recessive trait.

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WATARI, S., N. KATAGIRI, Y. HAGIYAMA and K . TSUGE, Department of Orthopedic Surgery, Hiroshima University, School of Medicine,

congenital malformation. Its characteristic features are abnormal over-growth of fingers or toes with sometimes total hand and foot involvement. The deformities resulting from overgrowth are quite different from that of neurofibromatosis o r lymphangioma. They are apparently brought about by abnormal tissue growth which is considered to be a functional disorder. The authors experienced fifty-two cases of macrodactyly in the hands and feet. Operative treatments were indicated to reduce digital size. The most characteristic and interesting finding observed during the operations was hypertrophic, lipo-degenerative changes of the digital nerves. This is supposed to be important and have a close relationship to digital overgrowth.

2 . For the differential diagnosis of syubrachydactyly and the constriction band syndrome, x-ray measurements of the bilateral radial, metacarpal and phalangeal lengths is useful because length variation is less in the constriction hand syndrome. 3 . With the increasing severity of constriction, centrifugal shortening and defects tend to progress and digits take the form of congenital amputation. 4 . These processes can be demonstrated in stages by diagnostic x-rays. 5. Acrosyndactyly is considered to be immediately followed by shortening of the digits in the abnormal limb morphogenesis. In this respect, three combinations of deformities are proposed. 6 . Whereas the deformity seems to be confined to the digits of the hand, tarsal involvement can also be observed.

YAMAMOTO, M., H. TAKAHASHI, H. HORIE, I. IWASAKI, K. ITABASHI, K. NAKAMURA and A. NITOH, Department of Pediatric Surgery, and Department of Pathology, School of Medicine, Chiba University, and Kawatetsu Hospital, Chiba. Beals syndrome (contractural arachnodactyly syndrome). Beals syndrome is a very rare disease characterized by long slim limbs, arachnodactyly, joint contracture, kyphosis, short neck and crumpled ears. The condition is transmitted as an autosomal dominant mendelian character. Case Report: The parents in this case study were cousins. Their first baby died of an unknown bone disease when he was two years old. The second child, a 3077 g male, was delivered by caesarian section at 38 weeks gestation. His Apgar score was 3 and he died 50 minutes after birth. There were no indications of chromosomal abnormalities but he displayed most of the clinical features of Beals syndrome.

TANABU, S . , Y. YAMAUCHI and I. TANGE, Department of Orthopaedic Surgery, and Department of Plastic Surgery, Juntendo University, Tokyo. Analysis of finger deformities in constriction band syndrome. The results of the analysis of the digital deformities in constriction band syndrome a r c . presented. The cases consisted of 2 4 males and 31 females. 34 cases ( 4 3 hands) had acrosyndactyly and 2 4 cases ( 2 8 hands) were without acrosyndactyly. The results are the following: 1. We were unable to find any extrinsic or intrinsic factors clearly responsible for the etiology of this anomaly.

MURAKAMI, M., K. MURAYAMA, S. SHINOZUKA, A. KUROSHIMA and H. OUCHI, Department of Obstetrics and Gynecology, Tokyo Women's Medical College, Tokyo. Three cases of repeated births of children with cephalic malformations. We have recently encountered three kinds of neonatal malformations; hydrocephaly, microcephaly, and anencephaly. Case 1: A second baby with hydrocephalus was born to a gravida 5 , para 2, abortus 3 woman. She had given birth to a baby with hydrocephalus. Hydrocephaly of the second baby was displayed by ultrasound at the second trimester of gestation and he was delivered by cesarean section at 38 weeks. A shuntoperation to the baby was not performed because there was no parenchymal tissue of the neonatal brain on cephalic CT scan. Case 2: A second microcephalic baby was born to a gravida 3 , para 2 , abortus 1 woman associated with hydramnios from the thirdtrimester. The mother had a previous microcephalic baby. The baby was still-birth at 32 weeks and the mother also showed hydramnios from about 30 weeks. Positive findings at autopsy were; hypertrophy of the cranial hones, almost complete closure of the cranial sutures, a severely hypoplastic brain, tetralogy of Fallot and other abnormalities. Case 3 : A baby with anencephalus was born to a gravida 3 , para 3 woman who had a previous anencephalic baby. The baby showed signs of only having anencephalus. Chromosomal analysis of the umbilical blood cells revealed a normal karyotype. In all three cases, there was nothing unusual about the family history.

SUZUKI, A., Y. FUKAMATSU, H. IINUMA and T. FUKUTA, Department of Obstetrics and Gynecology, Shinshu University School of Medicine. Matsumoto, Nagano. Agnathia associated with hydramnios.

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In utero diagnosis of agnathia is difficult and is often not recognized during prenatal examinations. A rare case of agnathia assoclated with hydramnios is presented. The infant was born prematurely at 32 weeks' gestation, in the obstetric clinic of the Shinshu University Hospital, to a 28-year-old gravida 1, para 1 (harelip), woman. Prenatal history was normal except for hydramnios which appeared at 25 weeks' gestation. At the time of delivery it was found that the facial features were distorted due to agnathia and a small oral orifice. An attempt to insert a catheter through the small oral orifice of the infant allowed only a few centimeters of catheter to pass with no material aspirated. The infant, whose Apgar score at birth was 1, died 20 minutes after birth. Postmortem examination revealed a 1430 g premature female with complete absence of the mandible (agnathia), a small oral orifice (microstomia), ears fused in the midline (synotia), a bifid tongue, and cleft palate. All other anatomical features were found to be normal. The amount of amniotic fluid was 4080 ml. The placenta was slightly enlarged for the gestational age, measured 22 x 22.5 x 2 cm, weighed 410 g, but otherwise was histologically normal. Agnathia, microstomia, and synotia are very rare birth defects of unknown cause. When presented as isolated conditions, they are due to a maldevelopment of the first pharyngeal arch. The hydramnios condition was secondary due to the swallowing difficulty of amniotic fluid because o f the absence of the mandible and the muscles of the oral pharynx.

OHTANI, T., M. HACHIYA, S. HIRATA, T. YAMAMOTO, A. HOSOE, and Y. NISHIMURA, Department of Pediatrics, Toyohashi City Hospital, Toyohashi, Aichi. Tracheal agenesis associated with truncus arteriosus; A case report. A female infant weighing 2000 g, was born after a 41 week gestation period. She developed severe generalized cyanosis soon after birth. Several attempts at emergency intubation were unsuccessful, and her condition deteriorated until her death 3 hours after birth. Necropsy revealed the absence of a trachea from the vocal cords to the carina. The newborn had a normal larynx and vocal cord. A tracheo-esophageal fistula with an inner diameter of 1.5 nun was also found at the level of the carina. Her lungs were of normal size and no excessive accumulation of lung liquid was noted. An associated anomaly was truncus arteriosus. Among the series of 96 cases with anomalous trachea and/or esophagus collected by Kluth in 1976, only 2 were reported to have tracheal agenesis. This case, however, is the first report of tracheal agenesis associated with a tracheo-esophageal fistula and truncus arteriosus. Rare anomalies reported in the present case are a valuable adjunct to the study of organogenesis of the trachea and esophagus.

OHAKI, Y., Y. SASAKI and K. ITOH, Department of Pathology, and Department of Thoracic Surgery, Kanagawa Children's Medical Center, Yokohama, Kanagawa. Thymic abnormalities in newborns with cardiofacial syndrome. Cayler et a1 (1967) first described the clinical manifestations of cardiofacial syndrome. It consisted of an asymmetric face, when crying and congenital cardiovascular anomalies. Four autopsy cases of cardiofacial syndrome with thymus gland abnormalities have been seen during the past 12 years. In two cases, the thymus was absent except for dysplastic thymic tissue detected by a semiserial histological examination of the anterior cervical region. In one case, thymic tissue showed some glandular structure but very few lymphoid cells. Both infants had high susceptibility to infection. We examined the thymic tissue in 100 cases of congenital cardiovascular anomalies without asymmetric crying face and did not find any thymic anomalies. Developmental failure of the pharyngeal arch around the sixth week of gestation called the Di-George syndrome appears to have common features with the genesis of the cardiofacial syndrome. Infants with asymmetric crying face should be routinely examined for immunological deficiencies.

SEKIGUCHI, S., Department of Anatomy, Tokyo Women's Medical College, Tokyo. Comparative study of the cardiovascular system in two cases of thoracopagus twins. Different types of conjoined heart (cardio.. pagus) were recognized in two cases of thoracopagus twins. The twins were 10 month old females. One case had a conjoined heart that was classified as a double heart with a common atrium, four ventricles and three atrioventricular valves. Another case had two divisible hearts, partially conjoined in the atrium and was classified as a divided heart. Both hearts were parallel to the sagittal plane of the fetus. Left superior vena cava, hypoplasia of the pulmonary truncus and a ventricular septa1 defect were recognized as common anomalies in the two cases. It seems difficult to speculate the degree of fusion of the cardiovascular system from the external form.

NAKAO, T., T. MOROHASHI, H. SHIMAZU, R. IIZUKA and T. OIKAWA, Department of Obstetrics and Gynecology and Department of Pediatrics, Keio University School of Medicine, Tokyo. Antenatal diagnosis of congenital A-V blocks and treatment with a pacemaker.

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Fetal arrythmia is not as rare as was previously believed. Advanced diagnostic techniques have identified arrythmia in 1.9% the patients that visited our OB-GY department. Two congenital A-V block cases were diagnosed before delivery with ultrasonic doppler and direct ECG. Both are alive because a pacemaker was attached just after birth. Both mothers were primigravida and had no special history. They visited our clinic when their fetuses were at 33 weeks and 25 weeks of gestation. Fetal heart rates were 100 110 bpm and 60 - 70 bpm. The ultrasonic doppler tests detected two waves showing independent cycles and different from the maternal cycles. Ultrasonic tomograms (electric scan) assumed that their waves were derived from the atrium and ventricle, and fetuses were diagnosed as having a complete A-V block. Patients were followed carefully and at 38 weeks and 39 weeks of gestation, they were delivered by cesarean section. Immediately, pacemakers were attached to the newborns.

and was admitted for evaluation, An electroencephalogram demonstrated hypsarrhythmia. A pneumoencephalogram and CT scan showed agenesis of the corpus callosum and enlargement of the third ventricle. The ocular fundi showed multiple punched out patches. Studies for infectious agents including toxoplasma, rubella, cytomegalic inclusion disease, herpes and syphilis, gave negative results. At 2 years and 5 months, she displayed severe mental retardation with delayed psychomotor milestones, although seizures were controlled by anticonvulsants. It was suspected that organic solvents contributed to the pathogenesis of this disorder.

KUROKI, N., S. SASAKI and Y . OGINO, Department of Plastic Surgery, St. Marianna University, School of Medicine, Kawasaki, Kanagawa. Cervico-oculo-acoustic syndrome. In 1952 Wildervanck reported a combination of the Klippel-Feil syndrome with deaf mutism, paralysis of the external rectus of one or both eyes and retractio bulbi. In this syndrome, a number of other congenital malformations pay be present such as pterygium colli, facial asymmetry, external ear defects and auricular deformities. Most of the patients were female and hereditary factors appeared t o be contributory, as the families of the patients included cases of deaf-mutism, anencephaly and rachischisis. The possibility of X-linked dominant inheritance was considered. We are reporti@g a male patient showing typical Wildervanck (cervico-oculoacoustic) syndrome with facial asymmetry, microtia, meatal atresia, torticollis and bilateral coxal luxation.

RIN, K., T. TANAKA, S. YOSHIWARA, N. TSUKIMOTO and T. KOHZUKA, Department of Pediatrics and Department of Ophthalmology, Toranomon Hospitai, Tokyo. Aicardi infant born to a mother exposed to organic solvents. Aicardi syndrome is characterized by the association of agenesis of the corpus callosum, infantile spasms, specific chorioretinal lacuna, and mental retardation. It is generally agreed that this syndrome is a sex linked, dominant trait. A female infant was born to a 40 year-old mother, who had worked with organic solvents at a paint manufacturing plant five hours each day for the first 8 weeks of pregnancy. At 2 months of age, the infant developed convulsions

OHTA, S., T. NARUTO, K. TANAKA, M. GOTO, T. YAMANO, Y. ABE and M. SHIMADA, Department of Pediatrics, Shiga University of Medical Science, Ohtsu, Shiga. A case of primidone embryopathy associated with barbiturate withdrawal syndrome. Primidone (PRM) embryopathy is a rare condition characterized by multiple anomalies associated with maternal primidone therapy. PRM and phenobarbital (PB: derivative of PRM) blood levels of a male newborn, considered to have PRM embryopathy were determined. Case report: A male newborn was born after a 38-week gestation period to a 25-year-old primigravida. The newborn's mother had a history of seizures starting at 9-years of age subsequent to a traffic accident. Her seizures were controlled mainly with PRM in daily doses of 2000 - 3500 mg. No complications were noted during pregnancy. The newborn's weight was 2300 g. Jitteriness, hypertonus and tremor were noted immediately after the delivery. He had a hirsute forehead, thick nasal root, long philtrum and sparse hair which were typical for PRM embryopathy. Blood levels of PB were 90.92 ug/ml at 24 hrs, 70.99 ug/ml at 48 hrs, and 62.07 ug/ml at 72 hrs of age. PB disappeared from his serum 19 days after birth. Blood levels of PRM were 20.87 ug/ml at 24 hrs, 7.53 pg/ml at 48 hrs and 0.07 lig/ml at 6 days of age. While babies of barbiturate withdrawal syndrome are often treated with PB, this patient was treated without using PB or other sedatives. His subsequent development was normal.

HIRAKAWA, M., T. KOJIMA and A. KITA, Department of Plastic and Reconstructive Surgery, and Department of Pediatrics, Jikei University School of Medicine, Tokyo. A case of popliteal pterygium syndrome. We report a case of popliteal pterygium syndrome, which was complicated with popliteal

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webs, and various malformations of the face, limbs and external genitalia. The patient was a 44 day-old female, born after 40 weeks gestation. Her birth weight was 2900 g. Popliteal webs, cleft lip and cleft palate, a cord-like structure in the buccal cavity, left palpebral fusion, club foot cum pes equinus, syndactyly, hypoplasia o f the nails, labia mojora agenesis, and clitoral hyperplasia were observed at birth. Webbing of the skin of both popliteal joints from the ischial tuberosity to the heel bone region caused flexing of the lower limbs. A membranous cord-like structure between the upper and lower jaws in the buccal cavity restricted the opening of the infant's mouth. Removal of the cordlike structure was carried out 80 days after birth and restored full function to the mouth. 110 days after birth, multiple 2 plasty was carried out on the popliteal webs of both legs. On the left leg, M. ischiosuralis, common in animals, was seen. 5 months after birth, the bilateral cleft lip was repaired. There have been 50 reports of this syndrome around the world since its first description by Trelat (1819). Until this case, only one other report of a popliteal pterygium syndrome was described in Japan (Negishi et a1 1981).

fistula (Gross C-type) and radial dysplasia was observed. Other malformations were recognized at birth, including cleft lip, cleft palate, microphthalmus, a low set and hypoplastic right ear, a hypoplastic right thumb. During corrective surgery, we found right pulmonary sequestration, malrotation o f the intestine, annular pancreas and Meckel's diverticulum. Chromosomal analysis of peripheral blood lymphocytes showed a normal 46,XX karyotype.

KUROKAWA, H., K. SASAKI and N. KAMITA, Department of Surgery, Takeda General Hospital, Aizuwakamatsu, Fukushima. Intestinal malrotation with Ehlers-Danlos syndrome in two patients. __ Two brothers, 25 and 28 years of age with Ehlers-Danlos syndrome were presented with similar clinical pictures consisting of hyperextensible skin and joints, dolichostenomelia, arachnodactvly, myopia, funnel chest, bulla and recurrent ileus. Some of the findings were characteristic of Marfan's syndrome, however the cardiovascular system and the lens were normal. At operation, marked hypermobility and friability of the bowel with perforation, intestinal malrotation, mechanical ileus by abnormal multiple hands or ligaments, intestinal diverticulosis, esophageal hernia, relaxatio diaphragmatica and inguinal hernia were seen. A skin biospy revealed an abnormal finding of elastic fiber. To our knowledge, this is the first report of malrotation with Ehlers-Danlos syndorome.

UCHIDA, T., I. ERA, K. USHIJIMA, T. ON0 and T. TOMODA, Department of Obstetrics and Gynecology, and Internal Medicine, Arao City Hospital, Kumamoto, and Department of Dermatology, Kumamoto University School of Medicine, Kumamoto. An autopsy case of a harlequin fetus. In February 1982, a 1920 g female harlequin fetus was delivered by caesarean section to a 19 year old primigravida after 3 4 weeks gestation. Apgar's scores were 8 or 9 at 3 minutes. There was no family history of ichtyosis, or consanguinity. The infant's appearance was grotesque, and her facial features greatly distorted; ectropion and chemosis were so extreme that the eyeballs could not be seen, and severe eclabium was present. The ear lobules were rudimentary and no external meatus of the auditory canal could he identified. Although the nose was flattened, the nares were patent. The surface of the skin was covered by thick, tough, yellow-gray plaques with deep crevices running both transversely and vertically. The hands and feet, which were fixed in a flexed position, were ischemic, hard, and waxy in appearance. Scalp hair was sparse and fine. Fingernails and toenails were hypoplastic. She died in respiratory distress on the 7th day of life. Light microscopy revealed the pathological changes limited to the skin. The skin showed uniformity in appearance with a marked thickening of the stratum corneum. The stratum corneum was punctured by sweat ducts and hair follicles. The stratum granulosum and stratum germinativum were essentially normal.

OZAWA, N., S . SATO, K. SOH, A . YAJIMA,

M. SUZUKI and R. OHI, Department of Obstetrics

and Gynecology, and Second Department of Surgery, Tohoku University, Sendai, Miyagi. A case of VATER association and other malformat ions. The VATER association was described by Quan and Smith (1973) as association of Vertebral defects, Anal atresia, Tracheo-Esophageal fistula, G n a l defects, and Radial dysplasia. A female infant with vertebral defects, imperforate anus with recto-vestibular fistula, T-E

MINE, Y., K. IWASAKI, T. MATSUO and T. IKEDA, First Department of Pathology, Nagasaki University School of Medicine, Nagasaki. A case of fetus in fetu. A 6-month-old male infant was examined for an abdominal mass. X-ray examination of the mass revealed ribs, vertebrae, iliac hones and long bones of the extremities. The mass was surgically removed from the left upper retroperitoneal cavity. It was enclosed in a membranous sac containing 100 ml of fluid. The mass weighed 389 and was covered by skin. Two lower limbs were attached to it. Histologically, the components of the tridermic tissue were confirmed. The following tissues were identified; skin, epidermal

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appendages, central and peripheral nervous tissue, bone, bone marrow, digestive tract tissue (esophagus, stomach, small and large intestine), an adrenal gland, a pancreas, striated muscle, adipose tissue and lymphoid tissue. In addition, a cluster of trophoblasts was observed, but no villi were detected.

No internal genitalia were palpable by rectal examination. BBT was biphasic, blood type was B and karyotype analysis was 46,XX in cultured blood. Urinary estrogen values was 16.1 glday, plasma progesterone and FSH values were 1.69 mg/l and 11.3 iu/l, respectively. IVP revealed a normal renal system. At laparoscopy, the absence of a uterus and the presence of normal, bilateral adnexa were confirmed.

YAMAOKA, S . , Y. OKANO, R. MURATA, M. KIN, K. TAKEDA, A. TANAKA, G. ISSHIKI and K. NARISAWA, Department of Pediatrics, Osaka City University Medical School, Osaka, and Department of Pediatrics, Tohoku University, Sendai, Miyagi. A case of V.B12 dependent methylmalonic acidemia. A patient with methylmalonic acidemia was found to display severe acidosis. A 2-yearold girl of consanguinous parents had hypotonia, an unbalanced diet, and showed growing and mental retardation since 8-months-old. She also had frequent episodes of metabolic acidosis since she was 11-months-old. Additionally, she displayed hyperglycinemia and methylmalonic aciduria. Methylmalonic acid CoA mutase activity was decreased i n skin fibroblasts, and it was corrected to normal levels in medium containing deoxyadenosylcobalamine, indicating that this was a case of methylmalonic acidemia responsive to V.B,,. Low protein diet therapy (2 g/kg/day) and oral administration of V.B,, were very effective. Decreasing methylmalonic acid in urine, weight gain and development were revealed. Computerized tomography of the brain showed an improvement of her brain atrophy.

MANO, T., M. GO., N. SUGANUMA, T. ASAI, M. FUJITA, 0 . NARITA and T. YASUMA, Department of Obstetrics and Gynecology, and Department of Ophthalmology, School of Medicine, Nagoya University, Nagoya, Aicbi. Nine cases of testicular feminization syndrome. The testicular feminization syndrome (TFS) is a type of male pseudohermaphroditism with characteristic clinical features. Essentially normal appearing women with females external genitalia, completely feminine habitus and normal female breasts lack internal female organs such as ovaries, fallopian tubes or a uterus. Instead, they have intra-abdominal testes. I n the last 10 years, 9 cases of TFS were seen in our clinic. Except for one case with 45XO/46XY mosaicism, all other cases had a normal male karyotype of 46XY. Two cases were an incomplete form of TFS withsome masculinization. There were 3 patients of TFS in one family with 2 of these patients being color blind (red-green).

IWAZAKI, K., A. FUJII, Y. KUROSHIPIA and M. INOUE, Department of Obstetrics and Gynecology, School of Medicine, Tokai University, Isehara, Kanagawa. Two cases of Mayer-Rokitansky-Kiister syndrome. Case 1: An 18 year-old unmarried female was referred to the hospital because of primary amenorrhea. She has a younger brother who is normal. The findings from a physical examination were unremarkable except for slightly immature breasts. Her external genitalia were normal but the absence of a vagina was recognised and only stringy internal genitalia were palpable upon rectal examination. RBT was biphasic, blood type was A and cytogenetic studies showed cells with 46,XX chromosomes in cultured blood. Urinary 17 KS and 17 OHCS values were 8.1 mg/day and 7.8 mg/day, respectively. IVP revealed an agenesis of the right kidney. At laparoscopy, only the presence of a left normal fallopian tube and ovary was confirmed. Case 2: A 27 year-old married female was referred to the hospital because of primary amenorrhea. She was the fifth of five children and the siblings were normal. The physical examination was unremarkable. Her constitution and external genitalia were normal. However, her vagina was 2 cm deep and the tip was blind.

TORIUMI, T., K. HONDA, Y. NISHI and N. TSUTSUMI Department of Obstetrics and Gynecology, National Ohji Hospital, Tokyo, and National Ohkura Hospital, Tokyo. A case of hydrocephalus associated with combined extra and intrauterine pregnancy diagnosed by ultrasonic real-time scanner. Combined intra- and extra-uterine pregnancy is very rare, and only one case out of 30000 cases was reported by Winer et al. In Japan, this abnormality has been reported in less than 50 cases up to now. Because of the difficulty in diagnosis, only a few cases can be diagnosed before operation. The remarkable technical advances of ultrasonic real-time scannershave allowed diagnosis of in utero abnormalities. We recently found two gestational sacs ( G S ) one outside and the other inside the uterus at the 11th week of gestation. The fetus with its heart movement in each GS, was observed. However, at the next examination, the coagulation echo was noticed in the cul-de-sac. After the left salpingotomy, the patient was progressing favourably, but the enlargement of the ventricles of fetal brain and development of Bip-D were found at the 33rd week of gestation. Therefore, therapeutic abortion was performed after diagnosing hydrocephalus.

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MATAYOSHI, K., T. OGAWA, Y. SUZUKI, T. KIYOKAWA, K. YOSHIDA and H. SOMA, Department of Obstetrics and Gynecology, Tokyo Medical College, Tokyo. Two cases of familial occurrence of hydatidiform moles. Few papers have reported the familial occurrence of complete hydatidiform mole (HM). Two cases of familial occurrence of HM are presented in this study. Case 1: HK, a 34 year old woman, had HM at 20 weeks of gestation after artificial abortion and normal spontaneous delivery. Curettages were twice performed. She became pregnant twice after the mole. Although the first one resulted in spontaneous abortion, the second continues up to now. OS, a younger sister of HK aged 33, had HM at 13 weeks of gestation. At that time, bilateral lutein cysts were found. After curettages, she recovered. After the mole, she delivered a normal baby. Case 2: AS, a 29 year old woman, had HM at the forth month of gestation. She was treated by evacuation and MTX (225 mg). After the mole, she had three babies. OK, a 26 year old sister, had HM at 12 weeks during her first pregnancy. Bilateral enlargement of her ovaries was observed at that time. Treatment included evacuation and MTX (375 mg) and transabdominal puncture using laparoscopy. Although the etiology of HM is still ambiguous, the familial occurrence of HM suggests a genetic predisposition at ovulation.

conventional banding techniques, may be a cause for the altered phenotype in a case with an apparently balanced structural abnormal.ity as in the parent with the normal phenotype.

KASAI, R., S . UCHIDA, N. KATAOKA, K. NARAHARA, K. KIKKAWA, S. KIMURA and H. KIMOTO, Asahigawa Jidoin Children's Hospital, Asahigawa, Hokkaido, Department of Pediatrics, Kawasaki Medical School, Kurashiki, Okayama, and Department of Pediatrics, Okayama University, Okayama. Paracentric inversions of the long arm of chrocases. _ _ is an exceedingly rare structural abnormality. We presented here two unrelated cases with paracentric inversion involving the long arm of chromosome 3. Case 1 is a nulliparous 31-year-old female, who has experienced 3 early trimester spontaneous abortions. The karyotype is designated as 46,XX,inv(3)(q25q27). The habitual abortion seems to be the result of production of duplication-deficiency gametes due to meiotic crossing-over within the inverted segments. Case 2 is a girl aged 2 years and one month. At the child's birth, the mother and father were 26 and 31 years old, respectively. She was delivered uneventfully at term, measuring 45 cm and weighing 2550 g. The parents and 3 sibs were phenotypically normal. She showed short stature, speech retardation, delayed closure of anterior fontanel, and short fingers and toes. Chromosome analysis revealed an apparently identical paracentric inversion of the long arm of chromosome 3 in the patient and father. The banding patterns of the inverted chromosomes 3 in the patient from prometaphases, however, demonstrated a duplication of band q263, and the patient's karyotype was 46,XX,rec(3),dup(q263),inv(3)(q12q29)pat. This observation suggests that aneusomic recombination, which is difficult to detect by the

AZUMI, J., H. SHIONO, M. KAMATA, M. UMETSU, S. MOTOYA and S. IMAMURA, Department of Legal Medicine, Sapporo Medical College, Sapporo, Hokkaido, Hokkaido Children's Hospital and Medical Center. Otaru. and Hokkaido Rvoiku Center, Sapporb, Hokkaido. Two cases- of t(6;18) and t(6;21). Case 1. The patient was a 1-year-old female infant of a 27-year-old mother and 29 year-old father. The female had been born after a normal gestation of 41 weeks and weighed 3090 g but showed signs of jaundice. Delivery was by aspiration. Jaundice increased for five days and the treatment with ultraviolet rays was started and lasted for 36 hours. She showed the following abnormalities: Mental and developmental retardation, hypertelorism, short philtrum and hypotonia. Chromosome analysis of the patient showed a 46,XX,t(6;18)(922;23) karyotype. Her parents were normal. Case 2. The patient was a 1-year-old female infant of a 28-year-old mother and 26-year-old father. No family history indicating congenital anomalies was present. Delivery was spontaneous after a gestation period of 39 weeks. The birth weight was 1930 g. Her Apgar score was 9 at birth. She now has difficulty controlling her head movement and sjtting up. She showed the following abnormalities: Mental and developmental retardation, thick eyebrows, posterior low hair line, webbed neck, cleft palate, scoliosis, and congenital heart disease (ASD + VSD). Chromosome analysis of the patient showed a 45,XX,-6,-2l,+t (6;21)(q27;qll) karyotype, Her parents had normal karyotypes.

OKA, S . , H. NAKANO, T. YOKOCHI, K. UEDA, A. SAITO and Y. IGARASHI, Department o f Pediatrics, School of Medicine, Hamamatsu University, Hamamatsu, Shizuoka, and Shizuoka Children's Hospital, Shizuoka. A case of lop- syndrome. A new case of deletion of the short arm of chromosome No. 10 was found in a patient whose kariotype was 46,XY,deI(lO)(p13). The patient, was born after 37 weeks of gestation, weighing 2420 g and measuring 46 cm in length. He had an Apgar score of 7 at birth. Delayed psychornotor development and cyanosis has been observed. At 2 years and 5 months of age, the patient could sit and had head control but standing, walking and speech were impossible. Congential anomalies detected were: small palpebral fissures, hypertelorism, depressed nasal bridge, hypoplasia of philtrum, high arched palate, low set and malformed ear, small auricle, stenosis of meatus acusticus externus, small mouth, wide set nipples, navel hernia,

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retentio testis, congenital heart disease (tetralogy of Fallot), simian crease and ulnar loops on eight finger tips. Both parents had normal karyotypes.

CHIBA, T., T. TANAKA, H. DODO, T. MATSUNO, S. ARITAKI, T. HONDA and K. KISHI, Department of Pediatrics, Tokyo Medical Collage Hachioji Medical Center, Department of Pediatrics, Tokyo Medical Collage, and Medical Research Institute, Tokyo Medical and Dental University, Tokyo. A case of a 14 ring chromosome. We encountered a case of a 14 ring chromosome, a rare disease so far found only in one patient in Japan and not more than 10 patients in Western countries. The patient was a female first presented to u s with afebrile convulsions 0 mo. Her karytotype was 46, XX,r(l4) at age 1 (p12 or 13; q 3 2 ) . Born as a first child during the 41st gestation week with the weight of 2770 g, she had no history of abnormality in utero or during delivery. There was no paternal or maternal family history of consanguineous marriage. A physical examination on the first visit revealed microcephaly, flat occiput, blepharophimosis, high arched palate and a short neck. The development of motor function was delayed, with the head held erect at age 3 mo, sitting at 10 mo, and walking at 1 YK and a half. Though she spoke 2 - 3 words at age 1 YK and 1 mo, her vocabulary increased very little after age 2. Electroencephalograms taken during the interval between seizures showed no clear epileptic discharge. Due to the administration of an anticonvulsant, generalized tonic and clonic seizures ceased to occur at age 1 yr and 5 mo. Upon endocrinologic examination, stimulation with insulin, TRH and LH-RH elicited a normal increase in cortisol TSH, PRI., LH and FSH serum concentrations, but had only a small effect on GH levels.

control, delayed motor development and speech. She has had seizures since the age of 12 and was placed on anticonvulsants, hut her condition has never been fully under control. Her IQ score was only 31. Examinations at the age of 14 revealed: a height of 152 cm, a weight of 50 kg, a high-arched palate, caf6-au-lait spots on the abdomen and recurrent acne vulgaris on her trunk. Her EEG presented paroxysmaI spike activity. Her dermatoglyphics patterns showed a loop radial (Lr) at the left hypothenar. None of these abnormal chromosomes were found in the other members of her family.

SAKIYAMA, T., Y. NOGUCHI, H. SHIMIZU, S. TAKAHASHI, T. KITAGAWA and Y. NAKAGOME, Department of Pediatrics, Nihon University Hospital, Tokyo, and Department of Human Genetics, National Institute of Genetics,
Mishimn. Shiniokn.
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AKAMATSU, Y., G. ANDO, A. IESHIMA, K. TAKADA, S. TAKASHIMA and K. TAKESHITA, Department of Pediatrics, Tottori Prefectural Hospital, Tottori, and Division of Child Neurology and Division of Neuropathology, Tottori University, Yonago, Tottori. An autopsied case with 46,XX, i(18q). Autopsy case reports of chromosome type 46, XX,i(lBq) are rare. A 3-day-old girl had respiratory distress, cardiac murnurs, scoliosis, mild muscle hypertonicity and multiple dysmorphic features: round face, epicanthus, low nasal bridge, lowset large ears, long philtrum, micrognathia, prominent occiput, short webbed neck, short sternum, small pelvis, overlapping fingers, single flexion crease of the fifth fingers and prominent calcaneus. Dermatoglyphic studies 0 arches. The newborn had a revealed 1 recurrence of hyperthermia and died of status epilepticus ard cardiac failure at 88 days of age. A pathological examination showed cardiovascular anomalies (coarctation of the aorta, pulmonary stenosis and polyvalvular anomalies), costovertebral anomalies (10 ribs, fused ribs and hemivertebrae), maIrotation of the bowel, Meckel's diverticulum, mild hydronephrosis and cerebellar hypoplasia. Cytogenetic analysis was performed using peripheral blood and skin fibroblasts. The karyotype of the propositus was confirmed to be 46,XX,i(18q) by G-banding techniques. Karyotypes of the parents were normal. This case has characteristic features of both trisomy 18 and 18p- syndrome. However, costovertebral dysplasia is a rare feature in either trisomy 18 or 18p- syndrome.

bisatellited derivative of chromosome 15 (inv dup 15) is presented. In previous reports of the aberration on chromosome 15, retardation of psychomorter development and epileptic seizure vithout striking physical malformations have been described. The patient was a 14-year-old mentally retarded girl. She was born to a 35-year-old mother and a 34-year-old father after 40 weeks' gestation without special difficulties during pregnancy and labor. She has a healthy elder brother, and her family history is negative for mental retardation, consanguinity, and congenital malformation. She had a failure t o thrive; had feeding difficulty, delayed head

KAMINAGA, S., S. SUNAGA and K. OKUYAMA, Department of Pediatrics, Faculty of Medicine, Showa University, Tokyo. A case of partial deletion of the long arm of chromosome No. 21: Clinical findings and erythrocyte superoxide dismutase-1 activity. A male infant was born after 35 weeks gestation and weighed 1560 g. He was the second child of a healthy 29-year-old mother and a 32year-old father. The parents were not consanguineous. The infant at birth had downward

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antimongoloid palpebral fissures, microphtbalmia, blepbarochalasis, a prominent nasal bridge, low set ears, a high arched palate, micrognathia and a prominent occiput. He also showed corneal opacities, generalized hypertonia, systolic murmurs, an umbilical hernia, bilateral undescended testes, and hypoplastic external genitalia. Chromosomal studies of the patient were performed in peripheral blood lymphocytes. The karyotype was 46,XY,de1(21)(pter + q.22.1 or 22.2). The karyotypes of parents were normal. Dermatoglyphic findings showed a ten arch configuration of the fingertips, t'axial triradii, absence of right c triradius and left d triradius. This patients' SOD-I activity (7.56 U/mgHb) was not different from age matched normal infants, whereas an increase of the enzyme activity, 1.41 times of the control level, was observed in patients with 21 trisomy. The result suggests the gene dosage effect of the SOD-I, and that the gene is localized on 21q 22.1 or more proximal portion.

H A M , T., T. NAKANISHI, N. TOKITA and Y. YONEYAMA, Sano-Kosei General Hospital, Sano,

Kleinfelter's syndrome with an XXYY chromosomal pattern in 1964, it has been suggested that there exists a close relationship between the extra Y chromosome and this skeletal abnormality. We have observed an 8-year-old Japanese boy with XYY chromosome who had skeletal and other multiple congenital anomalies. He was the second child born to a 26-yearold mother and a 24-year-old father. Delivery was uncomplicated after 41 weeks of an uneventful gestation. His weight at birth was 3,350 g. He did not speak until 15 months and also did not walk until 19 months of age. On examination at the age of 8 years, he weighed 26 kg and was 126 cm in height. His range of motion was limited in both elbows and forearms. Roentgenograms revealed congenital radioulnar synostosis. Other skeletal anomalies included platybasia, oxycephaly, craniosynostosis and separation of the odontoid process. Furthermore, several anomalies were observed such as partial dental defects, mental retardation, left undescended testis (operated on when 2 years old), mild deafness, mild disturbance of speech, tuberous sclerosis, left amblyopia and left oculomotor nerve paralysis (secondary squint and optic nerve atrophy). Chromosome counts showed all 50 cells obtained by lymphocyte culture had 47,XYY and were positive for Y-body.

rics and Gynecology, Hamada Hospital, Tokyo. newborn case of 45,X with hemihypertrophy. Case: K. N., a female was delivered by cesarean section as a result of placental dysfunction. The father was 36 and the mother was a 31-year-old gravida 2, pard 2, abortus 0. The mother's pregnancy course was normal. Their family history showed nothing unusual. The infant, after 38 weeks of gestation, weighed 1870 g, was 43 cm in height and had a head circumference of 35.5 cm. Right hemihypertrophy was seen shortly after birth. Lengths of upper humeral bones were 5.7 cm (rt) and 5.6 cm (lt); the ulnar bones were 5.25 cm (rt) and 4.85 cm (It); the femoral bones were 6.6 cm (both); and the tibias were 5.65 cm (rt) and 5.4 cm (It). A physical examination revealed frontal bossing (especially of the right forehead), micrognathia, low set ears, mild short neck, single creases on both palms, "shield chest" with widely spaced nipples and hypoplastic genitalia. Oral mucosal scrapings showed no "drum stick" nuclei. Chromosomal examinations revealed 45,X karyotype. At 30 hours after birth the infant had a hemoglobin of 26 g/dl, a hematocrit of 82% and a REX of 7.08 x l o 6 . At 6 days after birth, her LH was 61 mIU/ml, her FSH was 68 mIU/ml. 3 months after birth, her LH was 8.3 mIU/ml and her FSH was 22.7 mIU/ml. At 3 months of age, an EEG examination showed a bicentral sharp wave, and excretory urography showed no abnormal features. At 8 months of age, she weighed 5450 g, had a height of 61 cm, a head circumference of 41.5 cm and sitting with head control. At present she shows no abnormal neurological features. We could find no reports in the literature concerning the relationship between hemihypertrophy and 45,X karyotype.

IWATSUBO, T., H. SHIMIZU, K. NISHIKAWA, M. CHIGA, H. MACHIDA, K. OKADA and E. OBATA, Department of Pediatrics, St. Lukes International Hospital, Tokyo, and Department of Obstet-

SHIMADA, T., Department of Hygiene and Preventive Medicine. Yamaaata Universitv School of Medicine, Yamagata. Aneuploidies in Syrian hamster fetuses. A number of aneuploidic Syrian hamster fetuses, especially, autosomal monosomic fetuses were studied one day before birth. Fetal weight and external structure of aneuploidic fetuses were compared with karyotypically normal fetuses. Fifteen, 2-6 month-old male Syrian hamsters were irradiated with single doses of 250 or 500 rad X-rays. Irradiation was limited to the posterior third of the body. Males were mated from day 5 to day 74 after irradiation to untreated 2-9 month-old females. Seven untreated males were used as controls. Pregnant females were killed one day before delivery. All viable fetuses were analyzed cytogenetically by direct preparations of minced liver. Karyotype analyses were carried out to identify any extra or missing chromosomes following the procedures described by Popescu and Dipaofo. Eleven chromosomal anomalies were observed among 512 fetuses conceived by 109 mothers. One trisomic (trisomy 21) and 10 monosomic fetuses were found. Of the ten monosomies, four were XO syndromes, four were monosomies of C group chromosomes, one was monosomy 2 0 , and one was monosomy C with Dqf chromosome. WhereI

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as no gross malformations were observed under the dissecting microscope, a trisomy 21 and one of the monosomy C newborns had the lowest weights in their litters. The remaining aneuploidic fetuses were normal in weight. XO fetuses had normal ovaries and uterine horns. These findings suggest that there is no abnormal phenotypic expression of the numerical chromosomal change observed in Syrian hamsters.

TSURUZAKI, T., M. YAMAMOTO and G. WATANABE, Department of Hygiene and Preventive Medicine, Niigata University School of Medicine, Niigata. Maternal consumption of antipyretic analgesics produces chromosome anomalies in F1 rat embryos. Cytogenetic effects of antipyretic analgesics were examined in Sprague-Dawley rats aged 8-18 weeks. Female animals were given either aspirin o r acetaminophen orally for 2 weeks prior to conception. Doses were 0 (5% arabic gum s o l . ) , 125 or 250 mg/kg.b.wt. in the aspirin treated group and 0, 500 or 1000 mg/kg.b.wt. in the acetaminophen treated group. The treatment was continued until day 11.5 postcopulation (the day when copulation occurred On day 11.5 females was denoted as day 0.5). were sacrificed and the embryos were examined cytogenetically using a direct method. Frequencies of all chromosome anomalies among embryos were 0.5% (1/115), 5.1% (5/98) and 4.7% (4/85) in the 0, 125 and 250 mg/kg.b.wt. of the aspirin groups, respectively. Frequencies were 7.1% (7/98) in the 500 mg/kg.b.wt. and 4 . 8 % 15/104) in the 1000 mg/kg.b.wt. groups of acetaminophen treatment. A significant difference was found between controls and the 500 mg/kg.b.wt. acetminophen treated embryos (P=0.019). When the differences were analyzed by the type of chromosome anomalies, the occurrence of triploidies was not related to the drug treatment. In contrast, the aneuploid formation increased significantly with treatment for both drugs (control: 0% (0/115), aspirin 125 mg: 5.1% (5/98), aspirin 250 mg: 4.7% (4/85), acetaminophen 500 mg: 6.1% (6/98) and acetaminophen 1000 mg: 4.8% (5/104). Karyotypes of these aneuploidies were found to he all mosaics, consisting of monosomy/normal or trisomy/ normal cells. In conclusion, it is sugsested that maternal treatment with aspirin or acetaminophen induces aneuploid cells in the F I embryos.

KIKKAWA, K., K. NARAHARA, S. KIMURA and H. KIMOTO, Department of Pediatrics, Okayama University School of Medicine, Okayama. Cytogenetic studies of children with clinically idiopathic mental retardation. Chromosome studies using banding techniques were performed on peripheral blood lymphocytes from 80 children (49 males and 31 females) with clinically idiopathic mental retardation. The subjects ranged from 1 to 15 years in age, and had neither severe motor handicaps nor three or more congenital malformations. Chromosome abnormalities were found in 9 cases (11.3%). There were 5 cases with autosomal aberrations (47,XX,+inv dup(l5q); 46,XX,deI(Z)(q33q35); 46,XY,del(5)(q22q31); 46,XX,de1(13) (q21q22) ; 45 ,XY,t(13; 14)/46,XY, t(13;14)), 3 cases with sex chromosome aberrations (47,XXY; 46,XY/41,XXY; 46,Y,-X,+inv dup (X)(p22)), and one case with an unidentifiable small extra marker chromosome. The chromosome variations, found in 3 cases (3.8%), included lqh+, 16qh- and inv 9qh. Incidences of each variant did not differ from that in the Japanese general population. The results showed that chromosome abnormalities contribute appreciably to the etiology of clinically idiopathic mental retardation. Furthermore, it is remarkable that small interstitial deletions of autosomes represent the important abnormalities found in affected children.

HIHAM, M., M. ZENIYA and M. HIGUMSHI, Department of Hygiene and Preventive Medicine, University of Tokyo, 1st Department of Internal Medicine, Jikei Medical University, Tokyo, and 2nd Department of Health Science, Faculty of Medicine, Yamanashi Medical School, Yamanashi. Antibody-dependent, cell-mediated cytotoxic reaction in Down's syndrome. Antibody dependent cell-mediated cytotoxicity (ADCC) activities were studied in noninstitutionalized Down's syndrome children and appropriate mentally retarded controls by the microplate method. Down's syndrome children had significantly lower activities than did controls. After the incubation with human lymphocyte interferon (IFN-Y), enhanced ADCC activities occurred in both groups.

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FUKUSHIMA, Y., Y. KUROKI, A . HATA and N. NIIKAWA, Division o f Medical Genetics, Kanagawa Children's Medical Center, Yokohama, and Department of Pediatrics, Hokkaido Universitv School of Medicine. - Samoro. - Hokkaido. Cytogenetic investigation of 12 patients with Prader-Willi syndrome by a high resolution chromosome banding technique. Twelve patients with Prader-Willi syndrome (8 males and 4 females aged one month to 12 years old) were studied cytogenetically. Chromosome preparations were made from threeday cultures of peripheral lymphocytes. Prophase or prometaphase plates were collected with a modified ethidium bromide technique (Ikeuchi, 1979). Cultured lymphocytes were 0 ug/ml of ethidium bromide for treated with 1 two hours and with 0.02 ug/ml of colcemide f o r
L.

one hour prior to harvesting. Chromosomes were GTG-banded and then photographed. Prophase O K prometaphase banding analysis revealed in all 12 patients a deletion in the long arm (4) of chromosome 15, while a male infant had a non-Robertsonian 15q;15q translocation with a deletion in a 15q. 7 patients had the karyotype of 46,XY or XX,del(lS) (qll.Iq12), 4 patients 46,XY o r XX,del(l5) (qll.lq13) and one patient 45,XY,-15,-15, +t(15;15)(15qter-t15p11.2::15q13+15qter). The affected region common to all 12 patients was confined to a sub-band 15q11.2 (ISCN,1981). This suggests that the deletion of 15q is a case of the Prader-Villi syndrome. The high resolution banding technique is useful for the early diagnosis of Prader-Willi syndrome.

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