Injection of diethylstilbestrol on the first day of incubation affects morphology of sex glands and reproductive behavior of Japanese quail

Japanese quail eggs were injected with DES (0.9-1,000 micrograms) dissolved in 50-microliter of corn oil on day 1 of incubation. Higher doses of DES (250-1,000 micrograms) reduced hatchability to 37-33% compared to 61% for corn oil-injected controls. Lower doses of DES (0.9-125 micrograms) had no effects on hatchability. In a second study, eggs were injected with 0.9 or 1.9 micrograms of DES and the survivors were assessed up to 12 weeks posthatch. DES did not affect hatchability, but did increase mortality during the first 4 weeks posthatch. Females were affected more than males. At 10 days of age, open-field activity of some birds was examined. The acquisition and reversal of a visual discrimination task was studied at 6 weeks of age. DES had no effect on these measurements. Ten females from each group were chosen randomly to determine egg production over a single 28-day period beginning at 6 weeks of age. Exposure to DES blocked egg production in these birds. The oviduct weights of 12-week-old females were decreased by 50%, but ovarian weights were not affected. Testicular weights were not affected. In a third study reproductive behaviors and social-dominance behaviors of males were markedly attenuated in birds exposed to 0.48 or 1.9 micrograms DES in ovo.     

Extrapolation of Rodent Studies on Amniotic Fluid Contaminatnts to Human Populations

If endocrine active chemicals (EACs) adversely affect human development, then there must be evidence of effects in animal models at properly scaled levels of exposure during pertinent sensitive periods as derived from quantified exposures of the human fetus. Our recent studies attempt to address both effects and exposures. First Study: Dams were gavaged from Gestation Day (GD) 14 through weaning on Post-Natal Day (PND) 21 with either corn oil alone (unexposed controls) or Low DES (0.5?mg/kg BW); High DES (5.0?mg/kg BW); GEN (15?mg/kg BW); GEN + DES (GEN at 15?mg/kg BW and DES at 0.5?mg/kg BW). No treatments affected duration of gestation, litter size or birth anogenital distance / birth body weights ((bAGD/bBW) or ratios of bAGD/cube root of bBW of pups of either sex. The ratio of weaning AGD to weaning body weight (wAGD/wBW) differed significantly between the control group and each of the estrogenic treatments in both sexes with larger wAGD/wBW values associated with each of the estrogenic treatments. Males exposed to High DES and GEN alone exhibited earlier onset of puberty. Only females in the low DES group showed an earlier onset of puberty. At 50 to 70 days of age, the ratios of male reproductive organ weights/body weight were unaffected by estrogen treatment in all groups except high DES which increased testicle weight and decreased epididymis, seminal vesicle, and prostate weights. Initial vaginal cycle lengths were affected only in the high DES group. Thus low doses of DES and GEN at levels comparable to the upper range of human exposure affect some but not all markers of sexual development. Second Study: Amniotic fluid samples obtained at routine amniocentesis between 15 and 23 weeks of gestation were assayed by gas chromatographic/mass spectrometric (GC/MS) analysis. The first group of amniotic fluid samples (n = 53) from 51 women were analysed for several xenobiotic EACs. Alpha-hexachlorocyclohexane, with a mean (± SD) concentration of 0.15 ± 0.06 (ng/ml), and p,p′-DDE, with a mean (± SD) concentration of 0.21 ± 0.18?ng/ ml, were detected in several specimens. Overall one in three amniotic fluid samples tested positive for at least one xenobiotic EAC. Another group of amniotic fluid samples (n = 62) from 56 women were analysed for phytoestrogenic EACs. The mean (± SD) concentration of daidzein and genistein in amniotic fluid were 1.14 ± 1.04 and 1.37 ± 1.00?ng/ml with maximum levels of 5.52 and 4.86?ng/ml, respectively. Overall, 26 and 34 of the samples had quantifiable levels of daidzein and genistein, respectively. Conclusions: One in three human fetuses were exposed to xenobiotic EACs and two of three human fetuses were exposed to phytoestrogenic EACs in utero. Our demonstrations that EACs have developmental effects in an animal model at levels of exposure that mimic those found in humans in North America during sensitive time-frames sustains concerns about potential adverse health effects of developmental exposures to EACs for the human fetus/neonate.     

Molecular differentiation of the mouse genital tract: altered protein synthesis following prenatal exposure to diethylstilbestrol

Exposure in utero to the synthetic estrogen diethylstilbestrol (DES) has been associated with the subsequent development of reproductive tract lesions in both women and experimental animals. Using the techniques of organ culture and two-dimensional (2-D) gel electrophoresis, the effects of DES on protein synthetic patterns were studied during fetal and neonatal development of the CD-1 mouse. The protein patterns, analyzed by comparing 2-D fluorograms after [35S] methionine incorporation at different developmental stages, were correlated with the histology at the same age. Several qualitative and quantitative changes in protein synthesis were observed after prenatal DES exposure. A protein, apparent by Day 14 of gestation, with molecular weight approximately 70,000 and pI of 5.8, was observed to be greatly diminished in all reproductive tract tissues exposed to DES during prenatal development. This alteration, induced in utero, persists through the early postnatal differentiation of the genital tract (17 days old). This protein may provide an early marker for alterations in normal reproductive tract function.     

Influence of growth and athletic training on heart and lung functions.

Studies were performed at rest and during exercise of varied intensity on 52 boys of pre- and post-pubescent age. Each age group consisted of boys who were engaged in a strenuous prolonged hockey training program; this group was compared with a matched control group who did not participate in a regular training program. Any differences observed in the measured lung functions could be explained on the basis of physical size. Relationship of pulmonary capillary blood flow (Qc) and pulmonary diffusing capacity (DLco) to oxygen consumption were similar to those reported for adults and no difference between the trained and control groups was found in either the pre- or post-pubertal aged boys. Similarily, the trained pre-pubertal boys did not differ significantly from their matched control group in respect to the relationships of heart rate (HR) and stroke volume (SV) for any level of oxygen consumption. In contrast, the post-pubertal trained boys had significantly lower HR and higher SV (P less than 0.01) at each level of work than the control group. These differences between the trained and control post-pubertal boys are consistent with training effects observed in adults. The lack of differences between the trained and control pre-pubertal groups was surprising. Whether the differences in the post-pubertal groups due to a detraining effect in the post-pubertal control boys (as compared to the pre-pubertal control group) or to a continued high level of physical activity during and after the on-set of puberty in trained boys cannot be answered by this study. The findings suggest the importance of high intensity exercise programs during the growth period of adolescence if the efficiency of the oxygen delivery system, and possibly its ultimate dimensions, are to be enhanced.     

The natural history of childhood asthma to adult life.

A randomly selected group of 331 children who had started to wheeze in childhood and a control group of 77 children were prospectively studied clinically and physiologically from 7 to 21 years of age. Most subjects improved during adolescence and about 55% of those whose wheezing had started before 7 years and stopped before adolescence remained wheeze free. Forty-five per cent of subjects who had apparently ceased to wheeze at 14 years had minor recurrences of wheezing between 14 and 21 years of age. Fewer than 20% of those with persistent symptoms in childhood had become totally wheeze free during adolescence, although there was amelioration in symptoms. Girls did less well during adolescence than boys, so that there was no longer an increased preponderance of boys with increasing severity of asthma. Normal growth was achieved in all grades despite the persistence of symptoms in many cases. At 21 years of age features of airways obstruction were often found during an interval phase, especially in those who had more persistent symptoms.     

The development of gender-related behavior in females following prenatal exposure to diethylstilbestrol (DES)

Animal research has shown that diethylstilbestrol (DES) present during the sensitive developmental periods of the hypothalamus and adjacent areas of the brain affects the development of sex-dimorphic brain structures and subsequent behavior. To test for corresponding behavioral effects in humans, 30 women with a history of prenatal DES exposure were contrasted with 30 unexposed women who had been referred to the same clinic for a colposcopic examination because of an abnormal Pap smear. Gender-role behavior of childhood, adolescence, and adulthood was assessed by means of a semistructured interview, the Gender Role Assessment Schedule-Adult, and the Bem Sex Role Inventory. The mothers of these women were interviewed about their daughters with the "mother form" of the same interview schedule. The results suggest that DES women show less orientation toward parenting than the controls. There were no consistent group differences in other domains of gender-role behavior.     

Complications of pregnancy in mice exposed prenatally to DES

Women exposed prenatally to diethylstilbestrol (DES) develop a variety of reproductive tract anomalies. Most of these anomalies have been replicated in strain CD-1 mice after similar DES exposure. Recently, impaired reproductive performance in DES-exposed women has been reported. To see whether the mouse model also replicates this defect, a study of reproduction was performed. Pregnant CD-1 mice were injected with DES and their female offspring were raised to breeding age. The latter were then exposed continuously to untreated males for a maximum of 4 months. Among 74 mated mice, 34 became pregnant and 11 of these pregnancies ended in abortion or stillbirth. Other anomalies encountered were: two fetuses with compressed heads, one of which seemed blocked from delivery by a vaginal adenocarcinoma; two uterine tumors, one of which was a teratocarcinoma; two teratomas located in uterine lumina; and two uteri containing placentas without embryos. Since the frequency of successful pregnancies in the DES-exposed mice was reduced below control levels to a degree similar to that reported for DES-exposed women, the validity of the mouse model has been confirmed for this characteristic.     

Physical exercise, body mass index, subcutaneous adiposity and body composition among Bengalee boys aged 10-17 years of Kolkata, India

A comparative study of 215 sedentary (no regular physical exercise undertaken) and 313 physically active (regular physical exercise undertaken) Bengalee boys aged 10-17 years was undertaken to investigate the differences in overall adiposity (body mass index), subcutaneous adiposity (skinfolds) and body composition (percent body fat, fat mass and fat mass index). Both groups had a similar age. The results revealed that boys who did not undertake regular physical exercise (NPE) had a significantly greater mean body mass index (BMI) compared with those who undertook regular physical exercise (PE); p < 0.001. The means for all the skinfolds as well as percent body fat (PBF), fat mass (FM) and fat mass index (FMI) were significantly higher among the NPE group. The percentile distributions of all these variables and indices were consistently higher among the NPE group. The results of ANOVA of physical exercise (PE = yes, NPE = no) and PBF, FM and FMI, with age as covariate, revealed that PE had a significant negative effect on all these measures of body composition even after controlling for the impact of age. The means in each case were greater among the NPE group. In conclusion, this study provided evidence that Bengalee boys, who undertook regular physical exercise, had significantly less adiposity compared with those who did not undertake regular physical exercise.     

Influence of prenatal steroid exposure on neonatal gonadotropin imprinting. Effect of single and combined hormone treatments on the chicken ovary

Chicken embryos treated with DES or AE at the 9th day of incubation showed postnatally an increase in ovarian mass, parenchymal zone thickness, follicle diameter and granulosa cell count. Perinatal gonadotropin treatment had a similar effect on these parameters in the control birds not receiving pretreatment, but had no influence on them in the birds pretreated with steroid. The tested ovarian parameters of the DES- and AE-pretreated birds still differed from the control at 6 weeks of age. Neonatal gonadotropin exposure induced imprinting in the birds not pretreated with steroid, to judge from an increased response (indicated by changed values of the tested parameters) to gonadotropin reexposure in adulthood. Similar, but considerably slighter changes were also shown by the steroid pretreated birds. It follows that in the birds prenatally exposed to AE or DES, supervening perinatal exposure to gonadotropin did not elicit greater changes than the prenatal exposure itself.     

中性粒细胞／淋巴细胞比值与药物支架内再狭窄关系

目的：探讨经皮冠状动脉支架植入术前中性粒细胞／淋巴细胞比值（Neutrophil-to-LymphocyteRatio,NLR）与患者药物支架（DrugElutingStent,DES）植入后发生支架内再狭窄（In—StentRestenosis,ISR）的关系。方法：回顾性分析2012年01月至2012年10月间有DES植入史并再次入院接受冠脉造影的92例患者（平均年龄65．22±9．73岁）的血液学检查、既往冠脉造影结果及再次冠脉造影结果。根据DES植入前的NLR的四分位点将所有入选患者分为3组：最低三分位组（n=31,0．91-1．83）,中间三分位（n=30,1．87—2．63）,最高三分位（n=31,2．66—4．67）,比较3组间ISR发生率有无差异。结果：再狭窄发生在最低NLR三分位的有3例（9．68％）,发生在中间NLR三分位组的有2例（6．67％）,而有4例（12．90％）发生在NLR最高三分位组（P=0．714）,3组之间的差异无统计学意义。结论：因稳定或不稳定型心绞痛而植入药物支架的患者,其术前NLR同支架术后ISR的发生无明显相关性。NLR不能作为DES植入后发生再狭窄的预测指标。     

In Utero Exposure to Diethylstilbestrol and Blood DNA Methylation in Women Ages 40–59 Years from the Sister Study

In utero exposure to diethylstilbestrol (DES) has been associated with increased risk of adverse health outcomes such as fertility problems and vaginal as well as breast cancer. Animal studies have linked prenatal DES exposure to lasting DNA methylation changes. We investigated genome-wide DNA methylation and in utero DES exposure in a sample of non-Hispanic white women aged 40–59 years from the Sister Study, a large United States cohort study of women with a family history of breast cancer. Using questionnaire information from women and their mothers, we selected 100 women whose mothers reported taking DES while pregnant and 100 control women whose mothers had not taken DES. DNA methylation in blood was measured at 485,577 CpG sites using the Illumina HumanMethylation450 BeadChip. Associations between CpG methylation and DES exposure status were analyzed using robust linear regression with adjustment for blood cell composition and multiple comparisons. Although four CpGs had p<10⁵, after accounting for multiple comparisons using the false discovery rate (FDR), none reached genome-wide significance. In conclusion, adult women exposed to DES in utero had no evidence of large persistent changes in blood DNA methylation.     

Infant leukaemias and cot deaths.

Infant leukaemias differ from childhood leukaemias in ways which suggest that when haemopoietic neoplasms combine fetal origins with rapid growth rates they prevent normal development of the reticuloendothelial system and thus cause the sudden death of apparently healthy babies (stillbirths or cot deaths). Cot deaths are commoner in boys and have a peak incidence during the first half of infancy-that is, during the period most affected by the switch from passive to active immunity. Babies born from July to December, who are intensively exposed to winter conditions form 1 to 5 months of age, are also at special risk. During this period more girls and more children born form January to June die of leukaemia; and within three months of birth an exceptionally high ratio of myeloid to lymphatic leukaemias has been replaced by a low ratio, which persists throughout childhood.     

Mother's age effect on the boys' anthropometric properties in the Region of Tuzla (Bosnia and Herzegovina)

Our objective was to establish if mother's age affects the anthropometric properties their 11 and 16 years old sons, and if the boys' BMI is satisfactory. Cross-sections, questionnaires, and statistics were used for data processing BMI (kg/m2) was presented according to the scale Quetelet's index nutritional status. Twelve anthropometric properties were researched. It was established that mother's age affects anthropometric properties in adolescent boys. The younger mothers' boys come to puberty with significantly higher mean values for most anthropometric parameters. In the course of adolescent jump, older mothers' boys have significantly higher anthropometric measures in comparison to their age boys delivered by younger mothers. After the completion of their intensive growth and development anthropometric measures in both category boys are equal or insignificantly higher in younger mothers' sons. Mother's age affects their sons' anthropometric properties of growth and development, particularly in prepuberty and puberty. After puberty that effect is not significant. According to BMI, the boys coming from the researched region make the group of underfed children. That is probably the outcome of bad living conditions, irregular and insufficient diet, stress, etc. in the course of four year aggression on Bosnia and Herzegovina.     

Childhood antecedents of schizophrenia and affective illness: social adjustment at ages 7 and 11.

OBJECTIVE--To investigate the social adjustment in childhood of people who as adults have psychiatric disorders. DESIGN--Subjects in a prospectively followed up cohort (the national child development study) who had been admitted as adults to psychiatric hospitals were compared with the rest of the cohort on ratings of social behaviour made by teachers at the ages of 7 and 11 years. SUBJECTS--40 adult patients with schizophrenic illnesses, 35 with affective psychoses, and 79 with neurotic illness who had been admitted for psychiatric reasons by the age of 28. 1914 randomly selected members of the cohort who had never been admitted for psychiatric treatment. MAIN OUTCOME MEASURES--Overall scores and scores for overreaction (externalising behaviour) and underreaction (internalising behaviour) with the Bristol social adjustment guide at ages 7 and 11. RESULTS--At the age of 7 children who developed schizophrenia were rated by their teachers as manifesting more social maladjustment than controls (overall score 4.3 (SD 2.4) v 3.1 (2.0); P < 0.01). This was more apparent in the boys (5 (2.6)) than the girls (3.4 (1.8)) and related to overreactive rather than underreactive behaviour. At both ages prepsychotic (affective) children differed little from normal controls. By the age of 11 preneurotic children, particularly the girls, had an increased rating of maladjustment (including overreactions and underreactions). CONCLUSION--Abnormalities of social adjustment are detectable in childhood in some people who develop psychotic illness. Sex and the rate of development of different components of the capacity for social interaction are important determinants of the risk of psychosis and other psychiatric disorders in adulthood.     

Altered activation/detoxication enzymology following neonatal diethylstilbestrol treatment

The effects of neonatal exposure to diethylstilbestrol (DES) on hepatic activation/detoxication enzyme levels in the adult rat were investigated. Neonatal exposure of male rats to DES (DES males) decreased the endogenous levels of UDP-glucuronyltransferase as compared to control males. Female rats exposed neonatally to DES (DES females) had higher endogenous epoxide hydrolase and glutathione transferase activity levels than control females. Adult animals treated neonatally with DES also had altered metabolic potential following exposure to 3-methylcholanthrene and phenobarbital. The DES males treated in adulthood with 3-methylcholanthrene had higher benzo(a)pyrene hydroxylase activities and lower UDP-glucuronyltransferase activity levels than did control males treated in adulthood with 3-methylcholanthrene. The DES males exposed in adulthood to phenobarbital had reduced cytochrome P-450 and glutathione transferase activity levels as compared with respective controls. The DES females treated in adulthood with 3-methylcholanthrene had lower benzo(a)pyrene hydroxylase and epoxide hydrolase activity levels than control females receiving 3-methylcholanthrene. The DES females challenged in adulthood with phenobarbital also had decreased benzo(a)pyrene hydroxylase, epoxide hydrolase, UDP- glucuronyltransferase, and glutathione transferase activity levels as compared with respective controls. Our results demonstrated that neonatal exposure to DES changed the endogenous levels of specific hepatic enzymes and altered the metabolic response of these adult animals to a carcinogen and a drug.     

An epidemiological study on the effect of N, N'-methylene-bis-(2-amino-1,3,4-thiadiazole) (MATDA) on outcomes of pregnancy

A retrospective cohort study on the effect of the suspected teratogenic pesticide N, N'-methylene-bis-(2-amino-1,3,4-thiadiazole) (MATDA) on outcomes of pregnancy was conducted. The exposed group (6,173 pregnancies) was composed of childbearing women who, during their gestation, consumed rice harvested from a field where MATDA had been applied. The control I group (10,145) came from an adjacent county where MATDA had never been used; the control II group (3,326) consisted of women who had been pregnant previous to the introduction of this pesticide in the same region as the exposed group. The validity of the survey was verified by comparability and data-checking studies. After adjustment for the calendar year, maternal age, and pregnancy order, no significant differences were found in rates of spontaneous abortion, fetal death and stillbirth, birth defects, as well as in sex ratio and birth weight between the study groups. The same result was found in the comparison of individual categories of malformation. The rates of the birth defects were 23.84, 21.49, and 22.78 per 1,000 live births for the exposed, internal, and external control groups, respectively. No dose-response relationship was revealed. The results were consistent with previous reports and indicated the difference between animal experiments and human exposures. Although MATDA is teratogenic in animals, it is clear that the pesticide does not adversely effect outcomes of pregnancy in humans as currently applied.     

Ethnic Differences in Subcutaneous Adiposity and Waist Girth in Children and Adolescents

The purpose of this study was to examine ethnic differences in adiposity as measured by sum of skinfolds (SKF) and waist circumference (WC) in children and adolescents, after statistical adjustment for the BMI and age. A cross sectional sample of 3,218 (55% white, 49% male) children and adolescents aged 5–18 years who participated in the Bogalusa Heart Study (1992–1994) were included in these analyses. Sex‐specific ANOVAs, adjusted for BMI and age, for each 2‐year age group compared measures of adiposity (SKF and WC) between ethnic groups. No significant differences in the proportions of children and adolescents who were overweight and obese by ethnicity or sex were found. Mean SKF in normal weight (P < 0.0001) and overweight (P < 0.0001) categories was higher for white than black children of both sexes. Across most age categories, white boys and girls had significantly higher SKF than black boys and girls, respectively (P ≤ 0.05). Across most age categories, white boys had significantly higher WC than black boys (P ≤ 0.05) with no difference in the girls, when adjusted for BMI and age. Measures of adiposity in childhood and adolescence were significantly higher in white children compared to black children, when adjusted for BMI and age. Throughout childhood and adolescence, white boys and girls had higher SKF and white boys had higher WC. Differences in adiposity between ethnic groups should be considered in disease risk assessment and stratification as they are observed even for a given BMI level.     

Adult outcome of normal children who are short or underweight at age 7 years.

OBJECTIVES--To evaluate the adult growth outcome (at age 23) of children who are short or underweight at age 7 years in whom no identifiable pathological cause exists for their poor growth. DESIGN--Longitudinal follow up of a birth cohort. SETTING--The national child development study (1958 birth cohort) of Great Britain. SUBJECTS--523 children with a height or a weight below the fifth centile at age 7. Of these, 70 (13.4%) were excluded because they had a longstanding illness that could account for their poor growth. The remaining 453 subjects, who were followed to age 23, provided the base group from which those with additional data, such as parental height, were obtained. RESULTS--55/174 (31.6%) boys who were short at age 7 became short men; 60/211 (28.4%) girls who were short at age 7 became short women. Among boys who were underweight at age 7, 46/160 (28.7%) were still underweight at age 23, while 61/200 (30.5%) girls underweight at age 7 became underweight women. Having short parents did not increase the probability of being small as an adult. Children with delayed puberty were as likely to remain small as those in whom puberty was not delayed. CONCLUSIONS--One in three normal children who was short or underweight at age 7 became a short or underweight adult. This informs the management of short children and may be valuable when prolonged growth hormone treatment for short stature is being considered.     

Brief communication: Emergence chronology of permanent teeth in Nigerian children

This study measured the mean age, duration, and sequence of the emergence of permanent dentition in Nigerian children and compared the findings with other population groups. The cross‐sectional study involved 1,078 Nigerian children, aged 4–16 years old, from selected primary and secondary schools in the Ife Central local government area in Ile‐Ife, Osun State. In general, compared to boys, girls had an earlier mean age of emergence of all the permanent teeth. Children from high socioeconomic class had an earlier mean age of emergence for the maxillary incisors (6.43 and 7.58 years) and mandibular incisors (5.28 and 6.44 years) compared to children from middle and low socioeconomic classes, although socioeconomic effects were more mixed for premolars and molars. Compared to their counterparts in the USA, Australia, Belgium, and Iran, Nigerian children showed an earlier mean age of emergence of all the permanent teeth studied except for Pakistani boys, who had an earlier mean age of emergence of the maxillary premolars and second molar. Poorer economic status has been associated with delayed dental development; however, when compared to other populations, the Nigerian children in this study have earlier mean emergence ages than children from wealthier countries such as the USA and Australia. Am J Phys Anthropol 153:506–511, 2014. © 2013 Wiley Periodicals, Inc.     

Sexual Differentiation of Cognitive Abilities in Women Exposed to Diethylstilbestrol (DES) Prenatally

In nonhuman animals, prenatal exposure to androgens or estrogens enhances development of male-typical characteristics (masculinizes) and impairs development of female-typical characteristics (defeminizes). We investigated the hypothesis that prenatal exposure to the synthetic estrogen, diethylstilbestrol (DES), similarly masculinizes or defeminizes cognitive development in women. Forty-two DES-exposed women and 26 of their unexposed sisters were studied. No group differences were seen for abilities at which females excel on average (verbal fluency, perceptual speed and accuracy, and associative memory), for abilities at which males excel on average (visuospatial abilities), or for abilities that do not show sex differences (vocabulary, nonverbal intelligence). The time of prenatal exposure to DES correlated with visuospatial performance with later exposure associated with better performance. However, the subgroup of women exposed to DES late in gestation did not differ from unexposed women on these measures. Results support the conclusion that prenatal exposure to DES has little or no influence on cognitive development in women. However, they do not preclude other types of early hormonal influences on human cognition, such as prenatal influences of androgen or influences of androgens or estrogens during the early postnatal period.