bluuebunny/crossref_metadata_2025 huggingface dataset:
Relationship between Teratogenic Effects and Tissue Binding Pattern of Concanavalin A in Rat Embryos
Authors: NISHIDA Atsuyuki , KOBAYASHI Takashi , ASANO Yuzo , ARIYUKI Fumio , NAKAYAMA Yoshio
Published: May 2008

ABSTRACT This study was conducted to examine the effects of concanavalin A (Con A) on rat neurulation. Con A was injected intravenously to pregnant rats at a single dose of 5, 10, or 20 mg/kg between gestational days (GD) 7.5 and 10.0, and the embryos were macroscopically examined on GD 11.5. Con A induced a high mortality rate, the treatment with 10 and 20 mg/kg on GD 9.5 was completely lethal. Con A caused a high incidence of neural tube defects (NTD), the incidence in the 10 mg/kg on GD 8.0 and 8.5 groups and in the 20 mg/kg on GD 8.0, 8.5 and 9.0 groups being significantly higher than that in the control group. Histological evaluation revealed that Con A inhibited neural tube expansion and altered neural crest cell shape. In the Con A‐treated embryos, the stratified structure of the neuroepithelium was irregular and its extracellular space was expanded. In rat embryos from intact dams, the distribution of receptors for Con A was demonstrated histochemically on the cell surfaces around neurulation sites between GD 8.5 and 10.5, but not on GD 7.5. These findings indicate that Con A bound around the neurulation sites in rat embryos may disturb the cell‐cell communication required to form the neural tube, resulting in NTD.
Effects of N‐acetyl‐L‐cysteine on Teratogenicity of Cadmium in Mice
Authors: NAYA Masato , SHUTO Katsuichi , YASUDA Mineo
Published: May 2008

ABSTRACT Embryotoxicity and teratogenicity of cadmium (Cd) and modulation of its effects by N‐acetyl‐L‐cysteine (NAC) were evaluated in mice. Pregnant ICR mice were intraperitoneally injected with 3.5 mg/kg of CdCl2 on day 10 or 11 of gestation (vaginal plug = day 0). Pregnant mice were pretreated with 160 mg/kg of NAC intravenously 2 hours before dosing with CdCl2. Pregnant mice were killed on day 17 of gestation. Fetuses were examined for external malformations, especially limb malformations, cleft palate and abnormal palatal rugae. There was little difference in body weight gain of dams during the gestation period in the groups treated with NAC plus Cd as compared with the groups treated with Cd alone. Pretreatment with 160 mg/kg of NAC decreased the fetal mortality, incidence of cleft palate and abnormal palatal rugae induced by Cd on day 11. On day 10, pretreatment with NAC decreased the incidence of Cd induced abnormal palatal rugae. These results clearly indicate that NAC exerts protective effects against embryotoxicity and teratogenicity of Cd.
The Teratogenicity of Methamphetamine is Influenced by Housing Conditions of Pregnant Mice
Authors: YAMAMOTO Yoshiko , YAMAMOTO Keiichi
Published: May 2008

ABSTRACTOur previous study suggested that the teratogenic effect of methamphetamine (MAMP) may be influenced by the housing conditions under which drug‐treated pregnant animals were kept (Yamamoto et al., 1992). In the present study, pregnant mice were housed individually and their teratological data were compared with those obtained in the previous study where pregnant females were housed in aggregated groups. Pregnant mice were individually housed in a plastic cage and they received i.p. 11, 13, 15, 17, 19, or 21 mg/kg MAMP hydrochloride on day 8 of pregnancy (plug day = day 0). All the pregnant females treated with MAMP survived to term; this was in contrast to the high mortality rate of the animals which were treated with MAMP and kept in groups. No significant difference in the number of implantations and the resorption rate was noted between the control and treated groups. Malformed fetuses (externally, skeletally or viscerally) were encountered in the MAMP‐treated groups at prevalence rates of 0.6–3.2%, but the rates were not significantly different from the control value. The incidence of malformations was significantly lower in the groups kept in solitary conditions than in aggregated groups examined previously. The present study showed that crowding of pregnant females can potentiate the teratogenic effects of MAMP in mice. Some maternal physiological alterations caused by crowded environment such as stress‐induced glucocorticoid secretion and maternal hyperthermia may be responsible for the potentiated teratogenicity of MAMP in crowded housing conditions.
Induction of the Hydrocephalus by the lntraventricular Injection of Aurintricarboxylic Acid in Mice Exo Utero
Authors: TAMAGAWA Minoru , KEINO Hiromi , TANIGUCHI Masahiko , MORITA Jiro , NARUSE Ichiro
Published: May 2008

ABSTRACT Aurintricarboxylic acid (ATCA) was injected into the ventricle of mouse embryo on day 14 of gestation using exo utero method. Severe hydrocephalus and destruction of aqueductus cerebri were induced in the ATCA‐treated fetuses. Thinning of the cerebral cortex and widespread cell deaths in the periventricular wall of cerebrum, thalamus and hypothalamus were observed in the histological sections. The cell death was investigated immunohistochemically using an antibody against single‐stranded DNA. This antibody did not react to the dying cells, so it was suggested that the cell deaths were not apoptosis. TRPM‐2 gene expression was investigated in the hydrocephalic and normal brains on day 18 of gestation using dot‐blot hybridization analysis. However, the gene expression in the hydrocephalic brain did not increase. In the present experiment, intraventricular injection of ATCA on day 14 of gestation induced severe hydrocephalus in mice, and cell deaths in the periventricular wall were considered to be necrosis.
Nutritional State and Catecholamine Metabolism in Macular Mice
Authors: XU Gui‐Qin , YAMANO Tunekazu , SHIMADA Morimi , TACHIIRI Toshiharu
Published: May 2008

ABSTRACT Catecholamine and amino acid contents in the brain of the macular mutant mouse, a model mouse of Menkes kinky hair desease, were studied at days 4, 7, 10 and 13 postnatally. The contents of norepinephrine and epinephrine in the macular mutant mouse brain were significantly lower than those of controls at all ages. In contrast, the content of dopamine in the mutant mouse brain was significantly higher than that in the controls at days 4 and 7, and then it decreased rapidly to become less than that of age‐matched controls at days 10 and 13. Body weight gain of mutant mice was remarkably stunted, and weight loss was observed at day 7 of age. Body weight at day 13 postnatum was 56.1% that of the control. To examine the possible effects of malnutrition on the catecholamine metabolism in the mutant mouse brain, the contents of various amino acids in the brain and serum were measured. In the mutant mouse serum, tyrosine and phenylalanine, which play key roles in catecholamine metabolism, were significantly reduced compared to control, whereas only tyrosine showed a significant decrease in the brain. These results indicate that the rapid decrease in the catecholamine content in the macular mutant mouse brain is possibly influenced by the decrease in serum tyrosine subsequent to progressive malnutrition.
Radiation‐Induced Apoptosis and Developmental Disturbance of the Brain*
Authors: INOUYE Minoru
Published: May 2008

The developing mammalian brain is highly susceptible to ionizing radiation. A significant increase in small head size and mental retardation has been noted in prenatally exposed survivors of the atomic bombing, with the highest risk in those exposed during 8–15 weeks after fertilization. This stage corresponds to day 13 of pregnancy for mice and day 15 for rats in terms of brain development. The initial damage produced by radiation at this stage is cell death in the ventricular zone (VZ) of the brain mantle, the radiosensitive germinal cell population. During histogenesis of the cerebellum, on the other hand, the external granular layer (EGL) is also radiosensitive. Although extensive cell death results in microcephaly and histological abnormality, both VZ and EGL have an ability to recover from a considerable cell loss and form the normal structure of the central nervous system. The number of cell deaths to induce tissue abnormalities in adult brain rises in the range of 15–25% of the germinal cell population; and the threshold doses are about 0.3 Gy for cerebral defects and 1 Gy for cerebellar anomalies in both mice and rats. A similar threshold level is suggested in human cases in induction of mental retardation.

Radiation‐induced cell death in the VZ and EGL has been revealed as apoptosis, by the nuclear and cytoplasmic condensation, transglutaminase activation, required macromolecular synthesis, and internucleosomal DNA cleavage. Apoptosis of the germinal cell is assumed to eliminate acquired genetic damage. Once an abnormality in DNA has been induced and fixed in a germinal cell, it would be greatly amplified during future proliferation, bringing with it the risk of producing malformations and tumors. These cells would commit suicide when injured for replacement by healthy cells, rather than undertake DNA repair which is rarely complete. In fact they show very slow repair of cellular damage. Thus the high sensitivity of undifferentiated neural cells to the lethal effect of radiation may constitute a biological defense mechanism.
A Case of Multiple Pterygium Syndrome Associated with Chronic Idiopathic Intestinal Pseudo‐Obstruction Syndrome
Authors: IITANI Hidemi , KUSUDA Satoshi
Published: May 2008

Multiple pterygium syndrome (MPS) is a rare condition characterized by pterygia of the neck, finger, antecubital, popliteal, and intercrural areas. The present case study describes a male patient afflicted with this condition. This case has two unique features 1) the patient exhibited both mental and physical retardation at 2 years of age, a rare symptom of this disorder, and 2) the patient also had chronic idiopathic intestinal pseudo‐obstruction syndrome (CIIPS). Although the pathogenesis of both MPS and CIIPS is unclear, some features of the present case suggest a possible pathogenic mechanism.
In Vitro Developmental Toxicity of Aspirin and Its Major Metabolites on Cultured Fetal Mouse Palates
Authors: KOSAZUMA Tsuneo , CHOU Ming‐Jen , KAWAUCHI Saju , SHIOTA Kohei
Published: May 2008

Palatal primordia of day‐12.5 ICR mouse fetuses were cultured in a chemically‐defined serumless medium by a suspension culture technique, and the developmental toxicity of aspirin and its metabolites on in vitro palatogenesis was studied. Explanted fetal palates were exposed in vitro for 72 hr to 0.5‐2 mM aspirin (ASP), 0.25‐2 mM salicylic acid (SA), 0.5‐2 mM salicyluric acid (SUA), 1–2 mM 2,3‐dihydroxybenzoic acid (3DHB), or 1–2 mM 2,5‐dihydroxybenzoic acid (5DHB). After 72 hr culture, ASP at 2 mM and SA at 0.25 mM inhibited the growth and fusion of palatal shelves, and SUA at 1 mM prevented palatal fusion. On the other hand, 3DHB and 5DHB did not exert any significant toxic effects on cultured palates at concentrations up to 2 mM. Judging from the 50% inhibitory concentration (IC50), SA (IC50= 0.9 mM) was the most toxic of the 5 compounds tested, with a decreasing order of ASP (IC50= 1.5 mM), SUA (IC50= 1.6 mM), and DHBs (IC50= over 2 mM for both 3DHB and 5DHB). With respect to developmental toxicity, cultured fetal mouse palates showed the susceptibility to aspirin and its metabolites which is intermediate between the susceptibility of rat embryos in vivo and that of postimplantation rat embryos cultured in vitro. The significance of fetal organ culture for evaluating developmental toxicity of chemicals is also discussed.
Effects of Acetazolamide on the Development of Mouse Limb Buds In Vitro
Authors: WATANABE Toshiaki
Published: May 2008

Forelimb buds of mouse embryos were cultured in a serum‐free BGJb medium by a roller system. At the beginning of culture, acetazolamide was added to the final concentration of 10−5 M or 10−4 M. After 6 days of culture, no morphological abnormalities of the developing forelimb buds could be detected. In morphometrical analyses, the morphological scores of antebrachium and hand in the right forelimb of female embryos were 1.8 and 2.2 in the 10−5 M and 10−4 M acetazolamide‐treated groups, respectively, and 1.6 in controls. There was no difference in the size of the forelimb among these groups. Concerning cartilage skeleton, the incidence of bending of the ulnoradius was increased to 64–90% in 10−4 M acetazolamide‐treated group, which was different from the control (0.22%). In addition, there was a significant difference in the protein and DNA contents in only right forelimb buds between the acetazolamide‐treated and control groups. However, there were no differences in sex or laterality in the development of forelimbs and cartilage skeleton. No pathological changes in the cartilage skeleton and mesenchyme were observed in the acetazolamide‐treated forelimbs. From these findings, there is the possibility that acetazolamide per se may primarily interfere with the development of the forelimb in mouse embryos at the molecular level in vitro.
Palatal Slit and Cleft Palate in Rats Treated with Glucocorticoids II. Comparative Teratogenicity of Prednisolone, Triamcinolone Acetonide and Hydrocortisone
Authors: WATANABE Chiaki , ISHIZUKA Yasuo , NAGAO Tetsuji
Published: May 2008

The induction of abnormal palatal development by three glucocorticoids; prednisolone, triamcinolone acetonide and hydrocortisone was evaluated in rat fetuses. Pregnant rats were injected subcutaneously with either prednisolone (12.5‐100 mg/kg/day), triamcinolone acetonide (0.25‐2 mg/kg/day) or hydrocortisone (100 mg/kg/day) on days 14 and 15 of gestation. These females were humanely killed on day 20 of gestation and viable fetuses were inspected for their palatal abnormalities. The frequencies of cleft palate were significantly higher in the group treated with 100 mg/kg/day prednisolone (10.6%), and in the groups treated with 0.5, 1 and 2 mg/kg/day triamcinolone acetonide (8.6%, 26.1% and 58.3%, respectively) than the control frequency of 0%. Triamcinolone acetonide was 70 times as potent as prednisolone in inducing palatal slit, with ED50 value of 1.0 mg/kg/day and 70 mg/kg/day, respectively. Hydrocortisone showed no potentiality for the induction of cleft palate and palatal slit. Other developmental abnormalities including omphalocele and general edema, late resorption, and growth retardation were induced by triamcinolone acetonide and prednisolone. These findings indicate that triamcinolone acetonide has a significantly higher potentiality for the induction of palatal slit in rats, as well as in mice, compared to prednisolone and hydrocortisone.
Congenital Malformations and Deformations in Provincial Hospitals in Indonesia
Authors: KADRI Nartono , ISMAEL Sofyan , RAID Noersida , SURJONO Achmad , HARIANTO Agus , MUSTADJAB Immanual
Published: May 2008

A study of congenital malformations and deformations in the neonatal period in 14 hospitals consist of 13 provincial government hospitals as teaching hospitals and one private children and maternity hospital is presented. The incidence of anomalies in the range of 6–15 per thousand of total births. The incidence in boys was higher than girls. Based on the ICD‐10, among the 2,621 anomalies, the most frequent of malformations and deformations were those of the musculoskeletal system (19.6%), followed by the nervous system (18.8%), the digestive system (18.7%), the circulatory system (12.9%), and the cleft lip and cleft palate (8.2%). Corrective and reconstructive surgery were performed by the Subspecialistic Division of the Surgical Department.
Histological Alterations in Gestational Day 13 Rat Embryos from Albendazole‐Treated Dams
Authors: MANTOVANI Alberto , MACRÌ Caterina , RICCIARDI Claudio , STAZI Anna Velia
Published: May 2008

Gestational day (GD) 13 rat embryos from dams treated p. o. with albendazole (ABZ) on GD 10–12 were examined as an experimental model for the pathogenesis of embryotoxicity induced by benzimidazole compounds. Dosage levels were 0, 10, 20 and 30 mg/kg b. w. of ABZ. At 10 mg/kg b. w., most embryos appeared macroscopically normal. However, the histological examination of a representative sample showed subtle alterations including discrete necrotic foci in the lateral and rostral telencephalon, delayed differentiation of lens, increased wavyness of the notochord caudad to the forelimb bud and absent or poorly defined apical ectodermal ridge in the forelimb bud. At 20 and 30 mg/kg b. w. a sharp, dose‐related increase in embryolethality, developmental delays and macroscopic abnormalities were accompanied histologically by marked necrotic alterations of a number of embryonic tissues. The telencephalon and neural tube were severely affected, showing also a regeneration attempt through the disordered formation of tubules. Besides neuroectoderm, necrosis and tissue depletion were mostly evident in neural crest and mesoderm‐derived structures, such as branchial bars, limb buds and paraxial mesoderm. Lesions in the somites showed a head‐to‐tail increasing gradient; the sclerotomes were consistently more affected than the dermatomes. Wavyness of the notochord cephalad to the forelimb bud and fragmented/abnormal nuclei in the erythroblasts were also noted. Examination of embryos during in vivo organogenesis may be a useful tool for investigating the pathogenesis of embryotoxicity.
BUS/ldr, a Mutant Mouse Strain Exhibiting Abnormal Behaviors: Behavioral Similarities of BUS Mice and Chemically Labyrinthectomized Mice
Authors: HANAI Atsuko , TSUJI Keiichiro , OTANI Hiroki , MORIYAMA Kenji , SHOJI Ryujiro , YONEZAWA Satoshi
Published: May 2008

The behavioral abnormalities of homozygotes (bus/bus) of mutant bustling mice, BUS/ldr, were characterized in comparison with those of heterozygotes (+/bus) and of chemically labyrinthectomized mice. Homozygotes were 4‐fold more active than heterozygotes, as evaluated in an open field (11 × 17 cm) for 24 hr by means of Animex. Homozygotes revealed the lack of righting reflex and of head nystagmus on a rotating board, the frequent backward‐moving and the inability to swim, in addition to characteristic behaviors such as circling, head bobbing and head tossing. Bilaterally labyrinthectomized mice behaviorally mimicked BUS homozygotes: The behavioral patterns of operated animals were essentially the same as those of BUS homozygotes with respect to all reflex and locomotor indices employed, suggesting that some peripheral vestibular dysfunction, though unidentified yet, is attributed to abnormal behaviors of BUS mice. Unilaterally labyrinthectomized mice were not hyperactive, nor exhibited circling behavior.
Simple Methods for Objective Assessment of Sperm Viability and Motility with MTT Assay and Sperm Quality Analyzer (SQA) in Rats
Authors: HARA Hiroaki , TAKAI Ryou , TANAKA Naomi , MIZOGUCHI Keiji , TSUNO Tatsuya , IGARASHI Shin‐ichi , USAMI Masayoshi
Published: May 2008

To confirm the applicability of MTT assay and sperm quality analyzer (SQA) to the examination of sperm viability and motility in rats, epididymal spermatozoa derived from male rats (Slc:SD) treated with nitrobenzene at a dose of 60 mg/kg/day for 16 days were examined by these methods. Epididymal fluid was suspended in Dulbecco's modified Eagle's medium containing 25 mM fructose in ratio of 1:125. Sperm concentrations in the epididymal fluid were 2.48 ± 0.55 (× 106/μl, mean ± SD, n = 8) in the control and 2.20 ± 0.54 in the nitrobenzene group, and did not significantly differ between the groups. Both absorbance unit of the MTT assay (1.497 ± 0.406 vs. 1.084 ± 0.350) and SMI (sperm motility index) value of the SQA (204.0 ± 15.3 vs. 182.5 ± 21.6) were significantly decreased in nitrobenzene group. The results suggest that the examinations of sperm viability and motility with MTT assay and SQA are available for the reproductive toxicology in rats.
Identification of Mutations in the Gene Encoding the Fibroblast Growth Factor Receptor 3 in Japanese Patients with Achondroplasia
Authors: KITOH Hiroshi , NOGAMI Hiroshi , YAMADA Yasukazu , GOTO Haruko , OGASAWARA Nobuaki
Published: May 2008

Abstract We examined mutations in the gene encoding the fibroblast growth factor receptor 3 (FGFR3) in nine Japanese patients diagnosed as typical achondroplasia (ACH) and two patients with hypochondroplasia (HCH) using polymerase chain reaction (PCR) coupled with direct sequencing. In our present cases, equivalent to previously reported cases in France and the U. S. A., all mutations affected with ACH were the same nucleotide change, a G→A transition at position 1,138, resulting in the substitution of arginine for glycine at position 380 of the mature protein. Therefore, the gene mutation of ACH indicates no significant differences between the Caucasian and the Japanese. On the other hand, PCR products from genomic DNA of HCH did not show the same mutation.
Marshall‐Smith Syndrome: Report of a Case and Review of the Literature
Authors: ENDO Akihiko , JIN Yuko , MASUNAGA Ken , SHIMADA Masami , FUJITA Yukihiko , MINATO Michiyoshi , TAKADA Masaaki , TAKAHASHI Shigeru , HARADA Kensuke
Published: May 2008

ABSTRACT Marshall‐Smith syndrome is a rare disorder characterized by accelerated skeletal maturation, bullet‐shaped proximal and middle phalanges, dysmorphic facial features, and failure to thrive, and is often associated with mental retardation of variable degree. We describe an 8‐month‐old female with this syndrome, who has a hypoplastic corpus callosum and extreme upper airway obstruction requiring tracheostomy. Also, we review the previous reports of this syndrome since 1971 (Marshall et al., 1971).
A Common Mutation in the Tyrosine Kinase Domain of the Fibroblast Growth Factor Receptor 3 Gene in Two Japanese Patients with Hypochondroplasia
Authors: KITOH Hiroshi , YAMADA Yasukazu , GOTO Haruko , OGASAWARA Nobuaki , NOGAMI Hiroshi
Published: May 2008

We examined two Japanese patients with hypochondroplasia (HCH) whether they had mutations in the gene (FGFR3) encoding the fibroblast growth factor receptor 3 using polymerase chain reaction (PCR) coupled with direct sequencing. In both of our patients, a C→A transversion was detected in nucleotide 1659, predicting a substitution of asparagine for lysine at position 540 of the mature protein which corresponds a part of the tyrosine kinase 1 (TK1) domain. Our results suggest that a common mutation specific for HCH in the TK1 domain of FGFR3 exists among Japanese patients with HCH.
Fetal Laser Surgery Exo Utero in Mice*
Authors: NARUSE Ichiro , KEINO Hiromi , TANIGUCHI Masahiko
Published: May 2008

ABSTRACT  It has been very difficult to investigate mammalian embryogenesis. Whole‐embryo culture systems allow the observation of mammalian embryogenesis and enable the manipulation of the embryos, but it is almost impossible to culture fetuses at the late gestational period. The exo utero method does allow the manipulation of mammalian fetuses at the late gestational period. In this review, the detailed method of exo ufero surgery in mice is described, and the advantages and disadvantages of the applications of laser equipment for fetal surgery are discussed.

It has long been known that arhinencephaly is often associated with agenesis of the corpus callosum. The relationship between agenesis of the olfactory bulb and that of the corpus callosum is not fully understood. In this review, an example of fetal laser surgery exo utero was introduced, in which fissuration of the corpus callosum was induced after the destruction of the anlage of the olfactory bulbs in the mouse embryo. It was thus elucidated that agenesis of the olfactory bulbs induced the agenesis of the corpus callosum.
Carbamazepine‐Induced Cardiovascular Abnormalities in Chick Embryos
Authors: SONODA Tohru , FUKUNAGA Kazumi , TASHIRO Shinjiro , OHBA Ken‐ichi , SUGIMOTO Tohru
Published: May 2008

ABSTRACT  Carbamazepine dissolved in propylene glycol was administered to chick embryos in order to determine its teratogenicity in the cardiovascular system. A single dose of 3, 5, 7, or 10 mg was administered at 72 hours of incubation (Hamburger‐Hamilton stage 18). On day 10 of incubation, embryos were removed and examined for cardiovascular abnormalities. Cardiovascular abnormalities were noted in 37.8%, 54.8%, 76.9%, and 88.0% of live embryos in groups treated with 3, 5, 7, and 10 mg, respectively. Our experiment represented a statistically significant increase of cardiovascular abnormalities and showed a significant dose‐dependent increase. Infundibular ventricular septa1 defect was the primary cardiac anomaly observed in this study. A high percentage of this anomaly was accompanied by hypoplastic right 6th aortic arch artery and/or persistent left 4th aortic arch artery. A small number of double outlet right ventricle was found.
Developmental Analysis of Chlorambucil‐Induced Occipital Blebs in Mice
Authors: SEKIMOTO Hiroshi , HATTA Toshihisa , MORIYAMA Kenji , OTANI Hiroki , MORITAKE Kouzo , TANAKA Osamu
Published: May 2008

SUMMARY This study was designed to examine whether defects of the meningeal anlage and intracranial hydrodynamic overload are involved in the pathogenesis of secondary cranioschisis after the neural tube closure. Chlorambucil (CA) 3.0–4.0 mg/kg administered to pregnant mice on day 7.5 of gestation (E7.5) caused abnormal blebs at the occipital region of embryos on E13.5 in a dose‐dependent fashion compared with control. At blebs, the mesencephalic ventricle was dilated, particularly on the dorsal midline and posterior to the pineal invagination. Histology showed fewer mesenchymal cells and vessels, as well as thinner neuroepithelial tissue than in controls. Transmission electron microscopy revealed that the cytoplasm of mesenchymal cells in the bleb region contained abnormal electron‐dense deposits, whereas deposits or other subcellular abnormalities were not evident in either the neuroepithelium or the surface ectoderm of the affected region. We administered 3.5 mg/kg of CA on E7.5 and studied the embryonic development. Mesodermal hypoplasia appeared on E11.5, whereas blebs were initially recognized on E13.5. On E15.5 and E18.5, the meninges and calvaria were hypoplastic, although no apparent cranioschisis was evident. To examine whether increased intracranial hydrodynamic pressure plus defects in the anlage of the meninges and cranium induces secondary cranioschisis, we injected kaolin into the cerebral ventricle on E13.5 to induce hydrocephaly. These embryos developed exo utero and we examined the morphology on E18.5. Hydrocephaly was induced and the defects were more severe in the meninges and cranium than in embryos exposed to CA alone, although cranioschisis was not histologically confirmed. These results suggested that hypoplasia in the mesoderm during early organogenesis underlies later abnormalities of the meninges and cranium, which may be involved in the pathogenesis of certain types of cranioschisis.
Japan Pharmaceutical Manufacturers Association (JPMA) Survey on Background Control Data of Developmental and Reproductive Toxicity Studies in Rats, Rabbits and Mice
Authors: NAKATSUKA Toshio , HORIMOTO Masao , ITO Minako , MATSUBARA Yoshio , AKAIKE Masashi , ARIYUKI Fumio
Published: May 2008

ABSTRACT The Non‐Clinical Evaluation Subcommittee of the Drug Evaluation Committee, Japan Pharmaceutical Manufacturers Association (JPMA) collected historical control data from developmental and reproductive toxicity studies, which were conducted by the member companies and associated contract laboratories between 1986 and 1993. The data include spontaneous incidences of fetal morphological alterations and other observations made at terminal cesarean sections in rats, rabbits and mice. In addition, mating and natural delivery data in rats are also provided. There were strain differences as well as inter‐laboratory variations in the incidences of fetal visceral and skeletal alterations for both rats and rabbits. These inter‐laboratory variations were attributed to differences in the selection of observation parameters, observation criteria and classification of the findings.
Heat Shock Proteins in Normal and Stressed Mammalian Embryonic Development
Authors: WALSH David , ZHU Xiao‐Ou , GRANTHAM Jane , TAYLOR Rosanne , INOUYE Minoru , EDWARDS Marshall J.
Published: May 2008

The heat shock response is associated with a characteristic inhibition of normal transcription and translation with enhanced synthesis of a multigene family of Heat Shock Proteins (HSP), a common cellular mechanism to all living organisms including the most primitive species. Elevated body temperature or hyperthermia can result from many agents including fever, hot weather and heavy exercise. In more modern surroundings induced hyperthermia can be achieved by hot baths, saunas, specific drugs, electromagnetic radiation and ultrasound. Hyperthermia during pregnancy has been shown to cause a wide spectrum of effects in all species studied, including humans. The outcome depends on the duration and severity of the heat shock experienced by both the mother and the embryo and the stage of embryonic or fetal development. In severe exposures in utero embryonic death and resorption or abortion are probably the most common outcomes. In less severe exposures major or minor developmental defects can result, with the CNS a major target for its effects. Heat shock can interrupt cell differentiation at critical stages of organogenesis which may lead to apoptosis. All living organisms appear to have developed protective mechanisms collectively known as the heat shock response. Heat shock triggers the abrupt suspension in the normal protein synthesis and the concurrent induction of heat shock genes (hsp) and the synthesis of a set of multi‐gene protein families known as the heat shock proteins (HSP). Each hsp has at least two copies, one which appears to function in normal embryonic development (cognate) and another that is stress‐induced (inducible) and can result in acquired thermotolerance, offering some protection against further damage. The inducible HSP are usually activated at critical inductive stages of organ development, suggesting they play a major role in cell differentiation. The HSPs appear to protect cells through their chaperone functions by binding to adhesive sites on newly synthesised or heat damaged and partially unfolded structural and functional proteins. This prevents the formation of functionless aggregates. Heat shock transcription factors (HSF) bind to a heat shock element (HSE) and initiate hsp activation. Although this research has defined some pathways indicating how and why heat can cause some defects, a means of preventing them has not yet emerged.
Midkine, A New Heparin‐Binding Growth/Differentiation Factor: Expression and Distribution during Embryogenesis and Pathological Status
Authors: SUN Xue‐Zhi , FUKUI Yoshihiro
Published: May 2008

Midkine (MK) is a 13 kDa heparin‐binding growth factor found as a product of a retinoic acid‐responsive gene. MK is rich in basic amino acids and cysteine and its sequence is not homologous with any other proteins so far reported, so it is a new family of heparin‐binding growth factor. It has been found that MK exerts variety of biological activities such as neurite‐promoting, neuronal cell survival and differentiation‐inducing activities. MK is strictly expressed during the mouse embryogenesis; among the adult organs, it is detected only in the kidney. MK is also strongly expressed in a number of human carcinomas and specifically localized in senile plaques in the brain of patients with Arzheimer disease. More recently, it has been reported that MK is an important molecule regulating inflammation response and tissue repair. These results demonstrated that the relevance of MK not only in normal development, but also in processes leading to tissue repair or diseases. Increased MK gene expression is a common phenomenon observed in many human carcinomas, therefore MK is of significant interest in cancer biology. As a new growth/differentiation factor, many issues including the detailed sites and the precise time of MK expression, the exact cellular source which synthesizes and secretes MK, the signal transducing receptors for MK, the mechanisms underlying those developmentally regulated expression and its potential clinical significance still remain unknown. To elucidate the molecular mechanisms of MK action will lead not only to a deeper understanding of developmental processes, but also to the ultimate obtaining a key to diagnose and treat human carcinomas.
The Distribution of Tenascin in Rat Embryos with Normal Heart and Cardiovascular Anomalies Induced by Bis‐Diamine
Authors: IWASAKI Keisuke , IKEDA Takayoshi , OHTANI Hiroshi , MATSUO Takeshi , HIGAMI Yoshikazu , SHIMOKAWA Isao
Published: May 2008

The mesenchyme in the truncal ridge and aortico‐pulmonary (A‐P) septum in the developing heart expresses tenascin (TN), a component of extracellular matrices. We observed immunohistochemically the localization of TN with reference to fibronectin (FN) distribution in rat embryonic hearts at gestation days 12. 13 and 14 and compared the results with those in bis‐diamine‐induced cardiovascular anomalies. In control rat embryonic hearts, TN was distributed in the dorsal mesocardium and the distal part of the truncal ridge at gestation day 12, and in part of the conal and truncal ridges, A‐P septum and proximal wall of aortic arch arteries at gestation days 13 and 14. The conotruncal regions with TN expression may correspond to the areas where cardiac neural crest cells migrate in the heart. In bis‐diamine‐treated rat hearts showing conotruncal anomalies, TN was localized in incompletely fused and hypoplastic conotruncal ridges, but the intensity and area of staining were decreased suggesting a decreased neural crest cell migration. FN was diffusely distributed in both the normal and anomalous embryonic hearts, including atrioventricular cushion tissue in addition to the region expressing TN. TN was densely distributed in the fusing area of the conal and truncal ridges in comparison to FN, suggesting that TN may be mainly expressed during the time of fusion of conal and truncal ridges to form the A‐P septum in the embryonic heart. In conclusion, bis‐diamine may cause the poor development of neural crest derived tissue, subsequently bringing about hypoplasia of the A‐P septum and truncal and conal ridges.
Teratology and the Web: Homepage of the Journal ‘Congenital Anomalies’
Authors: MATSUI Kohji A. , YASUDA Mineo
Published: May 2008

This note presents current state and background of the homepage of Congenital Anomalies, the online version of the official Journal of the Japanese Teratology Society, and also introduce some of the teratology‐related web sites, in the hope that the usage of these and the other Internet tools will contribute greatly toward the advancement of teratology studies. Due to the cross‐disciplinary concepts and principles in the field of teratology, no single web site can provide exhaustive coverage of teratology. Nevertheless, there is indeed a great deal of useful information for a teratologist available on the Web. Proper usage of the Internet by exchanging and sharing valuable information can yield the mutual understanding and better collaboration, which in turn is indispensable for the overall measures of congenital anomalies.
Measurement of DNA Damage in ICR Mouse Embryos at Preimplantation Stage Using a Comet Assay
Authors: MARUYAMA Asako , KUSAMA Tomoko
Published: May 2008

Abstract Recent studies on embryos irradiated during the preimplantation period, especially 2 hours post conception (2 hpc), have shown that the sensitivities of external malformations as well as embryonic death were likely to be higher than that of mice irradiated at the stage of organogenesis. In this study, the embryos were irradiated with γ‐rays at 2 hpc and the DNA damage in embryos at 26 hpc was measured with a comet assay, that is, an alkaline single cell gel electrophoresis. The comet assay has been applied to single cells such as lymphocytes and bone marrow cells. We improved the procedure for measuring the DNA damage of two‐cell embryos (26 hpc embryos). The frequencies of cells without comet tails in embryos irradiated with 0.5 and 1.0 Gy decreased statistically significantly in comparison with those in non‐irradiated embryos. The comet assay could be useful in the measurement of DNA damage for embryonic effects during the preimplantation period.
A Comparison of Infants Affected with Congenital Abnormalities Born in the Inpatient Clinic or Treated in the Outpatient Clinic of the Same Hospital in the United Arab Emirates
Authors: HOSANI H. AL , SAADE D. , ZAKARIA M. , EDWARDS M.A. , CZEIZEL A.E.
Published: May 2008

Cases with congenital abnormalities were evaluated in the inpatient and outpatient materials of AI Mafraq Hospital in the United Arab Emirates between 1992 and 1994. The quality of data and diagnostic skill were appropriate. The baseline birth prevalence of total congenital abnormalities was 38.9 per 1000 which corresponded to the international figures based on nearly complete ascertainment. The cases of inpatient clinic may represent the general features of congenital abnormalities in the United Arab Emirates and particular clusters of different congenital abnormalities were not found.
Lack of Teratogenicity of 3‐Chloro‐4‐(Dichloromethyl)‐5‐Hydroxy‐2 (5H)‐Furanone (MX) in Embryonic Mouse Palate Culture in vitro
Authors: TERAMOTO Shoji , SHIMIZU Naoko , HOJO Hitoshi , KOBAYASHI Hiroko
Published: May 2008

3‐Chloro‐4‐(dichloromethyl)‐5‐hydroxy‐2(5H)‐furanone (MX) is known as a by‐product of wood pulp manufacture and a contaminant of chlorinated drinking water. Since our previous studies (Teramoto et al., 1998, 1999) demonstrated in a micromass in vitro test a strong inhibitory effect of MX on rat embryo cell differentiation, the potential teratogenicity was investigated in this study by using a suspension organ culture system. Twelve‐day mouse embryo palatal explants were cultivated for 72 hr in the MX‐containing medium at a concentration of 0, 1, 10, 100 or 300 μg/ml and examined for closure of the palatal shelves. All control explants showed almost complete closure of the palatal shelves. Similar results were also obtained in the MX‐treated explants at concentrations up to and including 100 μg/ml. Immunohistochemistry revealed no difference between the control and MX‐treated explants in distribution of PCNA‐and TUNEL‐positive cells in the palatal mesenchyme and medial edge epithelium, respectively. When the MX concentration was raised to 300 μg/ml, palatal shelves remained wide open. However, histopathology revealed extensive pyknosis of the mesenchymal cells and loss of the epithelium. These results may indicate that MX is cytotoxic against the mouse palate at a high concentration, and that it has no cleft‐palate inducing effects in mice.
The Safety of Penicillin V: Oral Penamecillin Use During Pregnancy The Importance and Limitation of Recall Bias
Authors: CZEIZEL A.E. , ROCKENBAUER M. , SøRENSEN H.T. , OLSEN J.
Published: May 2008

The purpose of the study was to check the human teratogenic potential of penicillin V: oral penamecillin treatment during pregnancy in the population‐based dataset of the Hungarian Case‐Control Surveillance of Congenital Abnormalities between 1980 and 1996. The pairs of cases with congenital abnormalities and their matched population controls without congenital abnormalities were evaluated, in addition the data of total population and of patient control groups as referents were compared with the data of congenital abnormality groups. In the three study groups there were 38,151 pregnant women who had newborn infants without any defects (population control group), 22,865 pregnant women who had newborns or fetuses with congenital abnormalities, and 812 mothers of informative offspring with Down syndrome (patient controls). In the population control group 2,246 (5.9%) pregnant mothers were treated with penamecillin, in the case group 1,597 (7.0%) pregnant women had penamecillin treatment while in the patient control group 63 (7.9%) mothers used penamecillin during pregnancy. The use of penamecillin treatments during pregnancy was higher in the total case (i.e., congenital abnormality) group, than in the total population control group. However, the highest use of oral penamecillin was found in the patient control group. The case‐matched population control pair analysis did not show a higher use of medically documented penamecillin during the second‐third months of pregnancy, i.e., in the critical period for most major congenital abnormalities. In addition, an association between medically documented penamecillin use during the second and third months of gestation and different congenital abnormality groups was not found when total population control group or patient control groups were used as referent. Thus, the reported higher use of penamecillin among the mothers of cases and patient controls can be explained mainly by recall bias. Thus, treatment with oral penamecillin during pregnancy did not indicate detectable teratogenic risk to the fetus.
Immunohistochemical Distribution of HNK‐1 and N‐CAM in Rat Embryos Treated with Bis‐Diamine
Authors: FUJINO Hidetoshi , NAKAGAWA Masao , SHIMADA Morimi
Published: May 2008

Bis‐diamine is known to induce congenital anomalies including cardiac defects, thymic hypoplasia and snout defects in rat embryos. Cardiovascular lesions consist of persistent truncus arteriosus, tetralogy of Fallot and aortic arch anomalies, which are often found in DiGeorge syndrome. In the the present study, we investigated effects of bisdiamine on cardiac neural crest cells, which may be important to the teratogenecity. A single dose of 200 mg bis‐diamine was administered to pregnant rats at 10.5 embryonic day (ED). Immunohistochemical studies were performed on embryos at 11.5, 12.5 and 13.5 ED using anti‐HNK‐1 and anti‐N‐CAM antibodies. The embryos at 11.5 and 12.5 ED showed the similar distributional patterns of either HNK‐1 or N‐CAM positive cells in control and bis‐diamine treated embryos. N‐CAM immunoreactivities in the splanchnic mesoderm were quite weakly detectable in the bis‐diamine treated embryos at 12.5 ED. Closure of the neural tube was delayed in the treated embryos at this period. Immunohistochemistry of the control embryos at 13.5 ED demonstrated a continuous chain of N‐CAM expression between the neural plexus near the foregut and the fourth aortic arch, while no apparent continuity of N‐CAM positive tissue was observed in the bis‐diamine treated embryos. These findings suggested that bis‐diamine reduced the amount of neural crest cells which appeared to participate in the aortic arch formation or conotruncal septation. The primary effect of bis‐diamine in the induction of various congenital anomalies including cardiovascular malformations may be an inhibition of the normal development of the neural tube and neural crest.
Dermatoglyphic findings in congenital clubfoot
Authors: Turhan A. Bülent , Atasu Metin , Paydak Ferhan , Kutlu Abdurrahman
Published: May 2008

ABSTRACT  The dermatoglyphics of the hand of 33 male and 17 female, a total of 50 patients with congenital clubfoot deformity (CFD) were compared with those of 250 male and 250 female, a total 500 control cases. The most remarkable dermatoglyphic findings observed in CFD were the decreased frequency of ulnar loops and the increased frequency of whorls on all fingers, the decreased frequency of ulnar loops on the left long finger and ring finger and the right thumb, long finger and little fingers, the increased frequancy of whorls on the left long finger and the right thumb, long finger, ring finger and little fingers, the decreased frequency of palmar IV loops and the increased frequency of ? loops and t triradii and the decreased frequency of plantar Î and V̂ loops and the increased frequency of the p and p″ triradii on the soles.
Rare association of polysplenia with anencephaly
Authors: Kohsa Kazuhiro , Miyabara Shinichi , Yonemitsu Nobuhisa , Toda Shuji , Sugihara Hajime
Published: May 2008

ABSTRACT  Anencephaly is usually observed without association of severe malformations of other organs. A case of polysplenia, which was experienced in a 20‐week‐old anencephalic female fetus, was reported. The families of the parents had no history of neural tube defects. There was no consanguinity between the parents. The fetus showed a typical anencephaly with absent cranial vault, exophthalmus, hyperplastic thymus and hypoplastic adrenals. There was an atypical polysplenia consisting of five small spleens, absent gall bladder, short pancreas, small intestinal atresia and Fallot tetralogy. No visceral heterotaxy was observed in bronchi, lungs, liver or stomach. Though no direct association has been recognized between anencephaly and polysplenia, recent progresses in the studies of human fetuses and animal experiments help explain the relationship between anencephaly and heterotaxy syndrome. The mother of the present fetus had no history of hyperthermia, drug take or diabetes mellitus. She also had no infections such as virus and toxoplasma before and during pregnancy. There were no clues that would elucidate the etiology of the present anencephalic fetus with polysplenia. Precise surveillance and records of such cases should be advisable.
Application of stereology to the central nervous system: estimation of numerical densities of neurons and synapses or neuron number
Authors: Fukui Yoshihiro , Bedi Kuldip S.
Published: May 2008

ABSTRACT Two unbiased stereological techniques, the disector and the fractionator have been introduced. Synapse‐to‐neuron ratios were calculated from estimates of the numerical densities of neurons and synapses in the rat cerebral cortex. These estimates were made using the disector method at the light and electron microscopical levels. Neuronal nuclei were as the counting unit for neurons and paramembranous densities for synapses. The fractionator method was employed to estimate the total number of cerebellar Purkinje cells in rats. We employed a five‐stage scheme to count the nucleoli of Purkinje cells. The cerebellum was first divided into uniform random slices, which were cut, in turn, into strips. The chosen strips were cut into pieces. A systematic selection of tissues was taken, and embedded together in the same mold. Subsequently the sections sampled were examined with a light microscope. All Purkinje cell nucleoli visible in these sections were counted.
Reproductive and developmental toxicity of triphenyltin chloride in rats.
Authors: Ema Makoto
Published: May 2008

ABSTRACT Reproductive and developmental toxicity of triphenyltin chloride (TPTCI) was evaluated in rats. Although no significant increase in the incidence of fetuses with malformations was observed following administration of TPTCI during organogenesis, a significant increase in the incidence of postimplantation embryonic loss was found. TPTCI during early pregnancy, especially on days 0–3 of pregnancy, caused implantation failure, i. e., preimplantation embryonic loss. The effects of TPTCI on the uterine function, as a cause of implantation failure, were determined using pseudopregnant rats. TPTCI was given on days 0–3 of pseudopregnancy and the decidual cell response was induced on day 4 of pseudopregnancy. Rats was sacrificed on day 9 of pseudopregnancy and the uterine weight served as an index of the uterine decidualization. A significantly lower weight of the uterus, which indicates the suppression of uterine decidualization, was found at the doses which induced implantation failure. These doses of TPTCI also caused a significant decrease in the progesterone levels, which indicates reduced ovarian function. These findings suggest that TPTCI exerts adverse effects on uterine decidualization correlated with the reduction in serum progesterone levels and these effects are responsible, at least in part, for the implantation failure induced by TPTCI.