Effects of the teratogenic folic acid antagonist, 9-methyl pteroylglutamic acid on uronic acid levels in fetal rat limbs

Experimental pregnant rats were placed on a transitory folic acid-deficient regimen, i.e. dietary restriction of folic acid plus the teratogen and folic acid antagonist, 9-methyl pteroylglutamic acid; a regimen resulting in multiple congenital skeletal malformations. Controls received a folic acid-supplemented regimen. Accumulation of a primary component of the extracellular matrix, glycosaminoglycans (GAG), was quantitatively evaluated by measuring uronic acid concentrations in fetal limbs. On days 16 and 18 of gestation, fetal limbs were removed, pooled from each litter, and prepared for uronic acid, protein, and DNA quantitation. The treatment results in (1) a decreased rate of protein and DNA accumulation prior to day 16 of gestation, the rate not being affected from day 16 to day 18 of gestation; (2) a trend toward an increased rate of uronic acid accumulation prior to day 16 of gestation, and (3) a decreased rate of uronic acid accumulation from day 16 to day 18 of gestation. Furthermore, our observations confirm our earlier suggestion that there is a decreased rate of accumulation of some nonprotein component(s) in fetal limbs obtained from folic acid-deficient mothers from day 16 to day 18 of gestation.     

Influence of folic acid on birthweight and growth of the erythroblastotic infant. III. Effect of folic acid supplementation.

Seventeen infants with severe (14) or moderately severe (3) erythroblastosis were given daily oral supplements of 2-5 or 5 mg folic acid from day 16 (average) to 3-2 months (average). Their rate of weight gain, expressed as weight centiles, was followed for 1 year and was compared with that of a very similar group of 34 erythroblastotic infants without folic acid supplements. By the end of the 4th month, just after stopping additional folate intake, the median centiles for weight had risen from the 40th to the 80th centile, while in the untreated control group they rose during this period from the 35th to the 50th centile. During the second half of the year both groups declined in weight centiles, the ''treated'' group ending up at the 50th centile for weight, while the control group fell to the 25th.     

Effect of folic acid supplementation on small-for-gestational-age infants born at term

The effect of 250 g folic acid (FA)/day or placebo given to 21 small-for-gestational-age infants born at term was studied during the first 3 months of life. The design of the study was double blind with random allocation. No randomization was performed in respect of breast-feeding or formula-feeding with a folate content of 60–70 g/l. No significant differences were observed in haemoglobin concentration, haematocrit, weight and length between FA-supplemented and non-supplemented infants. A negative correlation was demonstrated between gestational age and erythrocyte folate (E-folate) concentration at 1 week. E-folate content was higher when no supplement was given in breast-fed than in formula-fed infants.     

Ethanol and fetal nutrition: effect of chronic ethanol exposure on rat placental growth and membrane-associated folic acid receptor binding activity

Rat placental composition and specific folate receptor activity were measured at 20 days gestation in dams exposed to chronic high doses of ethanol (6%, vol/vol) throughout gestation and in isocalorically pair-fed controls. Ethanol-exposed fetuses were smaller (ethanol = 3.28 +/- 0.08 vs. control = 4.01 +/- 0.10 g, p less than 0.001), but their placentae were larger (experimental = 0.534 +/- 0.02 vs. control = 0.399 +/- 0.01 g, p less than 0.001). The increased weight appears to be secondary to hyperplasia as total DNA was increased while the wet/dry, RNA/DNA, and protein/DNA ratios were not different. Despite larger placentae, specific folate receptor activity was significantly reduced in the ethanol-exposed tissue, whether expressed relative to membrane protein, placental weight, or total placental binding. These results confirm that ethanol exposure is placentotoxic and suggest an additional mechanism by which ethanol may lead to intrauterine growth retardation; namely, decreased folate receptor activity.     

Effect of long-term carbamazepine therapy on serum lipids,vitamin B12 and folic acid levels in children

Serum triglycerides, total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), very low-density lipoprotein cholesterol (VLDL-C), vitamin B12 and folic acid levels were studied in 16 children with epilepsy who had been receiving carbamazepine (CBZ), and in 16 healthy children. Our purpose was to determine whether there was any effect of CBZ therapy on serum lipids, vitamin B12 and folic acid levels. Age ranged from 5 to 19 years (12.25 +/- 3.79 years) and 5.5 to 18 years (12.16 +/- 3.53 years) in the study and control groups, respectively. The duration of CBZ therapy in the patients was between 1 and 4.5 years (3.01 +/- 1.04 years). Serum CBZ level varied between 4 and 12 microg/ml (6.26 +/- 2.07 microg/ml). There was no statistically significant difference in serum triglycerides, TC, HDL-C, LDL-C, VLDL-C or vitamin B12. However, mean folic acid level was found to be lower in the study group than that of the control group (p < 0.05). Nonetheless, serum folic acid levels were within the normal range in all patients. Our study demonstrated that CBZ therapy does not affect serum lipids, vitamin B12 and folic acid levels, and may safely be used with regard to these parameters in children.     

Effect of carbamazepine therapy on homocysteine, vitamin B12 and folic acid levels in children with epilepsy

Objectives

To compare the levels of homocysteine, vitamin B12 and folic acid before and after 6 months of carbamazepine therapy and to correlate them with carbamazepine level at 6 months.

Design

Prospective comparative study.

Setting

Tertiary care centre in North India.

Participants

51 children (2–12 years of age) presenting with motor partial seizures.

Intervention

Carbamazepine (10–20 mg/μ/day) for 6 months.

Main outcome measure

Change in serum homocysteine, B₁₂, folic acid level.

Methods

Fasting venous samples were collected before carbamazepine therapy and after six months. Homocysteine was analyzed using homocysteine enzyme immunoassay. Vitamin B₁₂ and folic acid were estimated using electrochemiluminesence technique. Carbamazepine levels were measured at 6 months.

Results

Of the 51 children, 36 (males-21), were followed up and their data analyzed. Mean homocysteine level was 11.51±3.95 μmol/L at recruitment and 11.77±6.65 μmol/L at six months (P=0.785). At recruitment 6(16%) children had homocysteine level above 15 μmol/L which increased to 10(27%) at 6 months. Mean vitamin B₁₂ at recruitment was 292.1±111.2 pg/mL and 297.8±82.9 pg/mL at 6 months (P=0.764). Mean folic acid at recruitment was 9.98±3.45 ng/mL and 10.66±3.97 ng/mL at 6 months (P=0.358). There was no correlation between carbamazepine levels with homocysteine, vitamin B₁₂ and folic acid (P>0.05). There was no effect of age, sex or dietary pattern on homocysteine levels.

Conclusion

Hence 6 months of carbamazepine therapy did not cause significant change in serum levels of homocysteine, vitamin B₁₂ and folic acid.     

Effect of prenatal glucocorticoid on fetal lung ultrastructural maturation in hyt/hyt mice with primary hypothyroidism

Glucocorticoids (GC) and thyroid hormones (TH) accelerate fetal lung maturation. Though GC are used clinically, the mechanisms of GC-induced fetal lung maturity remain unclear. Prenatal GC increase fetal TH activity in humans and in animals. Thus, it is possible that increased fetal TH activity after prenatal GC plays a role in accelerating fetal lung maturation. However, this hypothesis has remained untested due to the lack of a suitable animal model. In the hyt/hyt mouse primary hypothyroidism occurs due to a point mutation in the beta subunit of the thyroid-stimulating hormone receptor of the thyroid gland, and it is transmitted in an autosomal recessive manner. We studied the effect of maternal betamethasone on fetal lung ultrastructure in hyt/hyt (hypothyroid) and Balb-c (euthyroid) mice. Hypothyroid mice were made euthyroid by T3 supplementation and mated to carry hypothyroid pups. Vehicle (n = 6) or betamethasone (n = 6) was injected intraperitoneally twice daily into the doe on days 16 and 17 of gestation. Fetal lungs on 18 days of gestation were subjected to ultrastructural morphometric analysis. The number of lamellar bodies per type II cell increased after betamethasone in Balb-c (2.10+/-0.31 vs. 3.43+/-0.37) and hyt/hyt (0.77+/-0.28 vs. 3.85+/-0.26) mice. The alveolar-to-parenchymal ratio was less in the vehicle-treated hyt/hyt (0.082+/-0.024) as compared with the vehicle-treated Balb-c (0.30+/-0.05) mice, while prenatal betamethasone increased the alveolar-to-parenchymal ratio in the hyt/hyt (0.227+/-0.034) but not in the Balb-c (0.26+/-0.04) mice. The glycogen-to-nucleus ratio was higher in betamethasone-treated hyt/hyt mice (1.46+/-0.20) when compared to vehicle-treated hyt/hyt (0.89+/-0.14) or Balb-c (1.01+/-0.17) or betamethasone-treated Balb-c (0.81+/-0.13) mice. Though tubular myelin was readily apparent in the airspace lumen of betamethasone-treated Balb-c mice, it was absent in betamethasone-treated hyt/hyt fetal lungs. We conclude that fetal thyroid plays an important role in accelerating some aspects of fetal lung ultrastructural maturation from GC stimulation.     

Effects of folic acid on preschool children''s appetite: Randomized triple-blind clinical trial

BACKGROUND: In recent years some Iranian pediatricians have used folic acid empirically as an appetite-enhancing drug in poor-appetite, low-weight children. Many parents have expressed their satisfaction with this treatment but until now no study has been done to confirm or exclude folic acid as an appetite enhancer. METHODS: In order to determine if complementary folic acid has any effect on preschool children's appetite, 61 3-5-year-old children (27 girls and 34 boys) whose weight/age and weight/height ratios were below the 25 centile and whose parents regarded them as having poor appetite were randomly assigned to receive either folic acid (1 mg/day) or placebo for 20 days. The primary outcome measures were any appetite changes during and 1 month after the end of the intervention, measured by means of a questionnaire completed by parents on the 20th and 60th days. Secondary measures were mean weight gain on the 30th and 60th days. RESULTS: Children who received folic supplement had a significantly better appetite on the 20th day than those who received placebo (mean difference of appetite score: 1.7; 95% confidence interval: 0.1-3.4; P = 0.04) and significantly more children receiving folic acid were reported to have increased appetite than the placebo group (P = 0.03). But 40 days after the end of the intervention there was no difference between the two groups. Weight gain did not significantly differ between groups. CONCLUSIONS: Although folic acid appears to improve preschool children's appetite, further study is necessary to determine whether it has any effect on growth as an adjunctive to management protocols of low appetite for undernourished children.     

Case report of a nutrition-induced anemia due to folic acid deficiency

Because of anaemia and dystrophy a girl was admitted to hospital. By bone marrow aspiration the diagnosis of a megalocytic anaemia was established. The determination of folic acid revealed a severe folate deficiency caused by education problems with consecutive malnutrition. Under folic acid replacement therapy and intensive talks with the parents the girl recovered from her illness and after change of nutrition she developed well without any further folic acid replacement.     

Latent coeliac disease in a child with epilepsy, cerebral calcifications, drug-induced systemic lupus erythematosus and intestinal folic acid malabsorption associated with impairment of folic acid transport across the blood-brain barrier

A 15-year-old boy with epilepsy and cerebral calcifications, treated with valproic acid, ethyl phenylbarbiturate and ethosuximide, was referred for drug induced systemic lupus erythematosus. Anti-gliadin (AGA) and anti-endomysium (EMA) antibody tests were both positive (EMA titre 1:50). Endoscopic duodenal biopsy showed intense chronic inflammation without villous atrophy or crypt hyperplasia. The child was discharged with a gluten-containing diet. The follow-up showed an increase in EMA titre (1:200) and the persistence of AGA. After 15 months, a second endoscopic intestinal biopsy showed flat mucosa and villous atrophy. Three serum folic acid determinations showed 1.8, 2.4, 2.0 ng/ml (reference range 2.5–16.9 ng/ml) prior to the two intestinal biopsies, but returned to normal levels (11.8 ng/ml) after a gluten-free diet and oral supplementation together. Two years later, the frequency of epileptic seizures was unchanged despite ongoing anti-epileptic treatment and a gluten-free diet. As cerebral calcification and epilepsy are reminiscent of the findings in congenital folate malabsorption, oral loading tests with 5 mg folic acid were carried out and showed impaired intestinal absorption and a defect in the transport across the blood-brain barrier. Low CSF folate levels (13.9 and 12.6 ng/ml, reference range 15–40 ng/ml) and an alteration in the CSF/serum folate ratio (1.43 and 1.16, normal ratio 3:1) were also found as well as increased levels of cystathionine both in CSF (40 μmol/l, reference range 18–28 μmol/l) and in serum (32 μmol/l, reference value <0.10 μmol/l). Conclusion Impairment of intestinal folic acid absorption with a defect in folic acid transport across the blood-brain barrier has been demonstrated in a case of epilepsy and cerebral calcifications associated with coeliac disease. Received: 12 November 1998 / Accepted: 29 February 2000     

叶酸缺乏对斑马鱼胚胎心脏发育影响的实验研究

目的 构建叶酸缺乏斑马鱼模型,观察叶酸缺乏胚胎的心血管发育异常表型,探寻预防叶酸缺乏导致心脏发育异常的最佳补充叶酸时期,并初步探讨叶酸缺乏导致心脏发育异常的机制.方法 利用二氢叶酸还原酶抑制剂甲氨蝶呤(MTX)以及使二氢叶酸还原酶基因(dhfr)表达阻抑两种方法 构建叶酸缺乏斑马鱼模型.并进行四氢叶酸挽救实验.在叶酸缺乏组胚胎发育不同阶段给予叶酸.观察并统计胚胎的心脏发育异常百分比、心脏发育异常表型、心率、心室收缩指数(VSF);利用荧光显微造影的方法 探查心脏流出道发育状况.采用原位杂交以及Real-time PCR的方法 检测各组胚胎vmhc、amhc、tbx5和nppa的表达情况.结果 与对照组相比,(78.00±3.74)%的MTX处理组胚胎以及(68.00±6.32)%的dhfr表达阻抑组胚胎存在心脏发育异常,包括心脏形态异常、心率和VSF下降以及心脏流出道发育畸形.给予四氢叶酸可挽救叶酸缺乏组胚胎的异常表型.在受精后8～12 h给予叶酸后,MTX处理组(20.20%±3.77%)以及dhfr表达阻抑组(43.40%±4.51%)胚胎的心脏发育异常百分比降低最显著,心脏发育异常状况改善最明显.MTX处理组胚胎以及dhfr表达阻抑组胚胎的vmhc和amhc表达水平正常;tbx5及nppa的表达水平明显下降.给予外源性叶酸后,tbx5和nppa的表达在MTX处理组胚胎以及dhfr表达阻抑组胚胎中增强.结论 本实验构建的叶酸缺乏模型中存在四氢叶酸水平下降.在胚胎发育早期中段给予补充适量的叶酸可以最有效预防叶酸缺乏导致的心脏发育异常.叶酸缺乏对心室以及心房肌球蛋白的分化无干扰作用.叶酸缺乏导致胚胎心脏发育异常与tbx5和nppa的表达水平下调有关.     

叶酸预防先天性心脏病的作用

目的了解孕早期增补叶酸预防先天性心脏病(先心病)的作用。方法调查1994年6月1日至1998年12月31日出生的先心病患儿的母亲和1996年8月1日至1998年10月31日出生的全部活婴的母亲孕前1月至孕后3月增补叶酸情况与先心病发生率的关系。结果(1)1996年8月1日至1998年10月31日出生的8332例新生儿的母亲妊娠初期增补叶酸与新生儿先心病发病关系显示,先心病患病率增补组为8.71‰(正规增补组为2.54‰),未补组为16.61‰。两组患病率差异经显著性检验,U=3.2584,P<0.01相对危险度(RR)0.5244,归因危险度(AR)0.0079,归因危险度比数(ARP)47.6%,归因危险度百分比(ARe)90.7%。RR值95%可信区间(CI)0.2992~0.7496。严重复杂先心病患病率未补组为增补组的5.3倍。(2)1994年6月1日至1998年12月31日出生的活婴中检得先心病221例,增补叶酸组62例、未补组159例,严重复杂先心病42例,不正规组为6例、未补组36例。新生儿期死亡18例、未补组17例,不正规组1例为21-三体综合征。先心病伴心外畸形11例,未补组9例。不正规组2例为21-三体综合征和先天性风疹综合征。结论孕前1月至孕后3月增补叶酸可减少先心病,尤其是严重复杂先心病的发生。     

叶酸预防先天性心脏病的作用

目的 了解孕早期增补叶酸预防先天性心脏病（先心病）的作用。方法 调查1994年6月1日至1998年12月31日出生的先心病患儿的母亲和1996年8月1日至1998年10月31日出生的全部活婴的母亲孕前1月至孕后3月增补叶酸情况与先心病发生率的关系。结果 （1）1996年8月1日至1998年10月31日出生的8332例新生儿的母亲妊娠初期增补叶酸与新生儿先心病发病关系显示，先心病患病率增补组为8．71％。（正规增补组为2．54％0），未补组为16．61％。两组患病率差异经显著性检验，U=3．2584，P〈0．01相对危险度（RR）0．5244，归因危险度（AR）0．0079，归因危险度比数（ARP）47．6％，归因危险度百分比（ARe）90．7％。RR值95％可信区间（C／）0．2992～0．7496。严重复杂先心病患病率未补组为增补组的5．3倍。（2）1994年6月1日至1998年12月31日出生的活婴中检得先心病221例，增补叶酸组62例、未补组159例，严重复杂先心病42例，不正规组为6例、未补组36例。新生儿期死亡18例、未补组17例，不正规组2例为21-三体综合征。先心病伴心外畸形11例，未补组9例。不正规组2例为21-三体综合征和先天性风疹综合征。结论孕前1月至孕后3月增补叶酸可减少先心病，尤其是严重复杂先心病的发生。     

Folate of pregnant women after a nationwide folic acid supplementation in China

Folate insufficiency during the periconceptional period increases the risk of neural tube defects (NTDs) in offspring, and folic acid supplementation substantially reduces the risk. Widespread large‐scale folic acid supplementation (0.4‐mg folic acid tablet) has been adopted as a main strategy to prevent NTDs in China since 2009. We examined folate concentrations in plasma and red blood cells (RBCs) of pregnant women and the factors associated with blood folate concentrations in a population with a high prevalence of NTDs in northern China. A cross‐sectional survey was conducted in 2014, and 1,107 pregnant women were recruited from 11 county or city maternal and child health centres across Shanxi province. Microbiological assays were used to determine folate concentrations. Factors associated with blood folate insufficiency were identified. The median (25th and 75th percentiles) folate concentrations were 28.4 (17.6, 45.2) nmol L^?1 and 1,001.2 (658.7, 1,402.5) nmol L^?1 in plasma and RBCs, respectively. According to the proposed RBC (906 nmol L^?1) concentrations for optimal NTD prevention, 42.4% participants had RBC folate insufficiency. Rural women had a higher proportion of folate insufficiency than urban women. Folic acid supplementation was the only factor associated with RBC folate insufficiency. A large proportion of women had RBC folate concentrations that are not optimal for the prevention of NTDs despite free access to folic acid supplements. Actions that aim to improve folic acid supplementation compliance are needed to reach the full potential of the nationwide folic acid supplementation programme in terms of NTD prevention.     

Juvenile granulosa cell tumor of the infant testis: case report with ultrastructural observations

This report describes the light-microscopic and ultrastructural features of a juvenile granulosa cell tumor of infant testis. Microscopic examination revealed a macrofollicular patterns simulating the preovulatory Graafian follicle and the juvenile granulosa cell tumor of the ovary. Ultrastructure confirmed three cell types: granulosa, theca interna, and externa, with occasional luteinized cells lacking crystalloids of Reinke. Charcot-Bottcher crystalloids were not detected, though rare cells contained a complex arrangement of filaments. An ultrastructural comparison was carried out with infant testes (2 cases), preovulatory Graafian follicle (1 case), juvenile granulosa cell tumor of ovary, adult granulosa cell tumor of ovary, and adult Sertoli cell tumor of testis and ovary. Ultrastructural similarities were noted between the present case and primitive Sertoli cells, preovulatory granulosa cells, and juvenile granulosa cell tumor of ovary. This may reflect the common histogenesis of Sertoli/granulosa cells from the common specialized gonadal stroma.