However, deleterious effects such as abnormal fetal growth, longer gestation period, increased birth weight, abortion, preterm birth and higher rates of neonatal mortality have been attributed to IVP. The aim of this study was to compare the influence of in vitro embryo production and artificial insemination (AI) on gestation length, complications with birth, birth weight, method of feeding colostrum, passive transfer of immunity, morbidity-mortality, and performance in Brahman calves.The use of bovine Whilst gestation length and birth weight were significantly increased in IVP-derived calves, no difference in weaning weight was observed between groups. The passive transfer of immunity (PT), was assessed in IVP (n = 80) and AI (n = 20) groups 24 hours after birth by determination of gamma-glutamyl transferase (GGT) and gammaglobulin activity as well as by quantification of the concentration of total protein in serum. No differences in passive transfer or incidences of dystocia and diseases at weaning were observed between groups. Birth weight, method of feeding colostrum and dystocia were not correlated with PT in either group.in vitro embryo production did not affect the health status, development, or passive transfer of immunity in Brahman calves.In this study,  This procedure allows for embryos to be obtained from genetically superior cows that are no longer able to produce offspring by conventional techniques or procedures. IVP increases the reproductive potential of the cows when compared with other biotechnologies such as artificial insemination (AI) [The use of bovine ion (AI) . Howeverion (AI) , it has ion (AI) ,3.Dystocia in newborn calves can cause asphyxia and so delay onset of natural suckling, negatively affecting the passive transfer of colostral immunoglobulins . Dystociin vitro embryo production and artificial insemination (AI) on gestation length, complications with birth, birth weight, method of feeding colostrum, passive transfer of immunity, morbidity-mortality, and performance in Brahman calves.The aim of this study was to compare the influence of Bos taurus indicus) (S: 23°04'34.7'' and W: 48°27'12.2'') located in Pardinho county in Sao Paulo, Brazil. The property comprises an area of 260 hectares, most part covered with Brachiaria sp. grass. The animals were raised under a semi-intensive system and were under permanent veterinary care.The study was conducted in a purebred Brahman farm , corn silage (3% BW), and 1% BW of a commercial concentrate .Regardless of gender, 100 Brahman calves were selected based on their method of production and divided into two groups during one calving season . Eighty calves were from IVP offspring (IVP group) and 20 were produced by artificial insemination (AI group). At birth, umbilical disinfection was performed with 5% iodine. Individual animals were identified by tattoo and cow tag earrings and dewormed with 200 μg/kg of body weight (BW) of doramectin subcutaneously. The calves were kept under natural grazing conditions with mineral salt licks and water All of the calves ingested colostrum during the 6 hours from birth, irrespective if by natural suckling, bottle feeding or nasogastric tube feeding. Calves were kept with their mothers until weaning . Over this period, clinical assessment of calves was performed and any disease treated.in vitro embryo production and artificial insemination techniques was obtained from two Brahman sires of an independently owned semen company. Follicular aspiration as well as in vitro embryo production and transfer were performed by a technician from a biotechnology company. Artificial insemination was achieved by a single technician.The cows used in our experiment were negative for brucellosis and tuberculosis and vaccinated against infectious bovine rhinotracheitis, bovine virus diarrhea, campylobacteriosis, leptospirosis and clostridiosis. Multiparous Brangus and Simbrasil cows were used as recipients for IVP embryos. Multiparous Brahman cows were artificially inseminated and the offspring used for the AI group. The semen used for both ad libitum and supplemented with corn silage (3% BW) and 1% BW of a commercial concentrate .One month before birth, dams were placed in maternity pens with mineral salt licks and water ®, Tokyo, Japan) with a 5 MHz sector transducer  attached to the transvaginal guide . Ovum pick-up was performed using an 18G needle under vacuum (500 mm Hg). The cumulus oocyte-complexes were collected into phosphate buffered saline medium  with 1% fetal calf serum (FCS) and 5 IU of heparin/mL. Prior to in vitro maturation COCs were assessed morphologically, and only those that had a cumulus morphology quality I and II [Cumulus-oocyte complexes (COCs) were obtained by ovum pick-up (OPU) from Brahman donors. Each donor was subjected to one OPU session using a SSD-500 ultrasound scanner  with 100 μL drops of modified TCM-199 medium with 10% FCS . This medium was supplemented with 5 μg/mL of FSH  and 50 μg/mL of LH , 1 μg/mL of porcine estradiol, 100 μg/mL of epidermal growth factor human (EGFh), 6.25 μg/mL of insulin, 22 μg/mL of pyruvate, and gentamicin. For in vitro maturation the COCs were incubated for 24 h at 39°C under humidified 5% CO2 in air.All selected COC's were ®, Lowell, USA) containing 100 μL Tyrode's albumin-lactate-pyruvate medium with 22 μg of pyruvate/mL, 6 mg of BSA/mL, 10 μg of heparin/mL, 20 μM of penicillamine/mL, 10 μM of hypotaurine/mL, and 2 μM of epinephrine/mL. Oocytes were inseminated with 2 × 106 sperm/mL and maintained at 39°C with 5% CO2 in saturated humidity for 18 to 24 h. Following insemination, putative zygotes were removed by successive pipetting. The in vitro culture was performed in 30 × 15 mm dishes with 100 μL Synthetic Oviduct Fluid medium [2, 5% O2 and 90% N2 at 39°C. All embryos were removed from culture on day seven of development and transferred to synchronized recipients.After maturation, oocytes were transferred to 30 × 10 mm dishes (CorningData were obtained regarding pregnancy technique (IVP or AI), gestation length, type of delivery , direct or indirect  colostrum feeding, and weight at birth, 30, 60, 90, 120 and 210 days after delivery. The animals were checked daily until weaning and more thoroughly when there was evidence of clinical alteration. Data on the presence and frequency of diseases were individually recorded.Blood samples from the calves were collected via jugular puncture using a 21G needle and serum tubes, 24 hours after birth. Samples were centrifuged and serum stored in plastic tubes at -20°C until tests. The protocol was approved by the Committee of Ethics and Animal Welfare of the Univ Estadual Paulista (UNESP), School of Veterinary Medicine and Animal Science, Brazil.®, Belo Horizonte, Brazil) and a SB-190 automatic analyzer  set at 405 nm length wave. Separation of the gammaglobulin protein fraction from serum was performed by electrophoresis in agarose gel  stained with amido black (0.1%) in 5% acetic acid [®, Barueri, Brazil). Total protein was quantified by direct biuret method [®, Belo Horizonte, Brazil). FPT was considered to have occurred when the calf had at least one of the following values: total protein (< 5.0 g/dL), gammaglobulin (< 1.0 g/dL) and/or serum GGT (< 100 IU/L) [Diagnosis of FPT was assessed by determination of gamma-glutamyl transferase (GGT) activity in serum, according to the kinetic-colorimetric method previously describe , with a tic acid . Samplest method  using th00 IU/L) .Data were analyzed by Chi-square or Fisher's exact test when variables were categorical. Due to the non-normal distribution of continuous variables, comparison between AI and IVP groups was performed using the nonparametric Mann Whitney U test. All correlations were conducted using the Spearman RHO test. The significant level of analysis was established at 5%.Bos indicus) [Gestation length of IVP offspring was significantly longer than AI offspring p < 0.001), with means of 294 ± 6.6 days and 287 ± 6.7 days, respectively  , suggestAs in other studies -15, birtMales were heavier than females at birth in a previous work . In our IVP individuals may have abnormal placental and/or fetal development, causing the so called large offspring syndrome, characterized by excessively large calves at birth, high perinatal mortality, hydrallantois and increased number of abortion . Calves in vitro and in vivo derived calves weighing more than 40 kg [Weight and size at birth, associated with the specific characteristics of the recipients and embryos, as well as with the number of births, were the main factors causing dystocia in Nelore recipients . Other aan 40 kg .Despite observing dystocia in 20% 16/80) of IVP births, i.e., one cesarean section and 15 births using obstetric chains; and in 5% (1/20) of the AI calves, no significant differences were identified between groups (p = 0.096) , gammaglobulin (< 1.0 g/dL) and/or serum GGT (< 100 IU/L). Failure of passive transfer was detected in 10% of the IVP offspring (8/80) and 20% of the AI offspring (4/20) without significant difference between groups. Only one AI-derived calf had lower GGT (22.3 IU/L), total protein (4.7 g/dL) and gammaglobulin (0.7 g/dL). Nevertheless, this animal did not experience disease and had a weaning weight of 208 kg, very close to the mean weight of the group.Bos taurus IVP-derived calves absorb large amounts of colostral immunoglobulins in order to acquire adequate passive immunity [Though passive transfer is essential for calf survival, several other factors may influence life and performance of the individual ,30,31. Aimmunity ,33. In aThe birth weight, method of feeding colostrum and dystocia were not related with FPT in any of the groups within the current study. Some studies have shown correlations between the increase in FPT and dystocia ,35; howeIn general, the low occurrence of FPT in both groups in this study was probably associated with the immediate and comprehensive assistance provided at delivery, thus minimizing the risk of fetal hypoxia and increasing the rate of feeding with colostrum during the first 6 hours after birth. Therefore, the FPT seems to be more related to fail in ingestion of colostrum by the calf rather than to a defective absorption of macromolecules through the intestinal epithelium .At least one disease was present in 40% 32/80) and 50% (10/20) of the IVP and AI groups respectively (p = 0.42) . As previously described  it is feIn Brazil, livestock production is characterized by large-scale extensive operations under grazing conditions mainly at north and central west of the country. Although the main reproductive strategies are artificial insemination and natural matting, the use of IVP increases each year in animals of high genetic value. With the objective of diminishing the influence of environmental factors, our experiment was performed through a single farm during one calving season. Moreover, this property actually represents an elite livestock Brazilian farm using IVP as the main biotechnology for reproduction and the number of animals used in this experiment accurately reflects the characteristics of the livestock segment in this country. Other authors ,24 obtaiBased on our results we can conclude that gestation period and birth weight were higher for the IVP offspring. However, weaning weight was similar between groups. The occurrence of dystocia, assistance for colostrum feeding, FPT and disease in calves were similar from birth to weaning in both groups. Birth weight, method of feeding colostrum and dystocia were not associated with FPT. Under these conditions, we can say that IVP technique did not have influence on health status and performance of Brahman calves or interfered with acquisition of passive immunity.The authors declare that they have no competing interests.APO and JPOF performed the clinical examination, laboratorial analysis, reviewed the literature and prepared the manuscript. AD performed the statistical analysis and prepared the manuscript. RCG reviewed the literature and prepared the manuscript. All authors read and approved the final manuscript.APO, JPOF and RCG are from School of Veterinary Medicine and Animal Science - Univ Estadual Paulista, UNESP, Botucatu, Sao Paulo, Brazil. AD is from School of Medicine of the UNESP, Botucatu, Sao Paulo, Brazil.


This study used data from recent Demographic and Health Surveys (DHS) to examine the impact of short or long preceding birth intervals on neonatal and under-five mortality. In order to minimize the effect of selection issues, we examined child mortality outcomes of the same mother, comparing short or long interval births against births with what had previously been considered optimal intervals.We analyzed 47 DHS datasets from low- and middle-income countries. For each dataset, we compared neonatal and under-five mortality of short preceding interval births  to reference interval births (24-<60 months) of a mother, using conditional logistic regression matching on the mother. We also conducted the same matched analysis for long  preceding interval births. These associations were then meta-analyzed. We also stratified the analyses by mothers’ completed fertility (fertility at end of reproductive period) to assess whether maternal characteristics highly correlated with completed fertility modify the association between birth interval and child mortality.Children with shorter preceding intervals had increased odds of both neonatal  and under-five mortality . When the associations were stratified by the mothers’ completed fertility, the impact of short intervals was greatly reduced or eliminated for low fertility mothers. In contrast, mortality associations became stronger for children of high fertility mothers. However, when the births of high fertility mothers were limited to birth orders 2-4, the associations were comparable to those of low fertility mothers. Longer preceding birth intervals had lower odds of mortality than reference intervals . This effect was also mediated by mothers’ completed fertility; there was a strong protective effect of longer birth intervals for the high fertility mothers but not for low fertility mothers.These analyses reproduced findings reported in previous literature that shorter birth intervals are associated with higher child mortality. However the negative impact of short birth intervals may only occur in high parity births. Reproductive health interventions that seek to lengthen birth intervals may have larger impact by targeting women with high parity. This finding is consistent with the concept of maternal depletion as the underlying cause of increased adverse child outcomes associated with shorter birth intervals. Short and long preceding birth intervals, or time between a birth and the subsequent birth, have previously been linked to adverse child outcomes-5. With In our analysis, we seek to estimate the association between birth intervals and child mortality, and the extent to which this association can be explained by physiology versus environmental causes. The literature reporting birth interval associations have mainly used cross-sectional data, and have sought to remove possible confounding effects such as maternal age, parity, and socioeconomic status from the birth interval-mortality association. To better extricate possible residual confounding from the true association, we use new analytic approaches on Demographic and Health Survey (DHS) data. We compare child mortality of risky interval and reference interval births of the same mother; by examining child mortality risk with a mother as the unit, we better control for residual confounding that cannot be addressed statistically. We also restrict the analysis to births that occurred when women were ages 18-<35 to eliminate age as a potential confounder rather than relying on statistical control. Finally, we only include women who were over age 35 at the time of the survey. This allows us to derive her fertility at the end or near the end of her reproductive period (completed fertility), which we will then use as a proxy for maternal characteristics that may possibly alter the association between birth intervals and child mortality.LiST). LiST is a software package that estimates the impact of scaling up interventions on reducing maternal and child mortality, stillbirths, and other outcomes such as stunting and wasting in low- and middle-income countries.[ LiST has been developed within Spectrum,[LiST to a family planning module; currently in the model, the number of pregnancies and resulting maternal and child deaths and stillbirths decrease as contraceptive use goes up. We seek to better understand the complex associations linking contraceptives, birth intervals, and their associations with child mortality, and update the LiST model with best available evidence.The overarching purpose of this paper and other papers in this supplement is to investigate the possible impact of contraceptive usage and its ensuing drop in fertility on the rates of child and maternal mortality. The findings will inform what biological links between reproductive health-related exposures and child mortality should be included in the Lives Saved Tool . We did not include surveys from Sao Tome and Principe and Ukraine due to low number of overall births and low number of high fertility births, resulting in 47 countries included in the analyses. If multiple Phase V surveys were conducted in a single country, we used the most recent survey.We first limited our analyses to children of mothers who were over 35 years of age at the time of survey. A separate analysis found that children of women with high fertility at the end of her reproductive period have higher mortality risk than those born to women with low fertility, regardless of birth order. This sugSurvival or death by 28 days  and five years of age  were recorded for the included births. Preceding birth interval is available for each child in the datasets, aside from first births. When looking at under-five mortality as an outcome, we excluded any births that occurred in the five years before the mother completed the survey, as we cannot fairly assess survivorship at age five. Similarly for neonatal mortality, we did not include any births that occurred in the month prior to the mother completing the survey.We examined two different cut-offs for short intervals (<18 months and <24 months) and two different cut-offs for long intervals (≥60 months and ≥72 months). 24-<60 month was used as the reference interval for all analyses, based on previous literature reporting this interval as optimal. We conducted conditional logistic regression for each of the aforementioned short or long intervals as the exposed group, matched on the mother. Then, the same analyses were conducted stratified by maternal completed fertility. Mothers who had four or fewer live births were categorized as low completed fertility and mothers who had five or more live births were categorized as high completed fertility, given the mothers were age 35 or older at the time of the survey. (All mentions of fertility hereafter refer to the mothers’ completed fertility.) In order to infer if short or long birth intervals make an impact on all births or only following a cumulative effect after numerous births, we re-ran the same analysis for high fertility mothers, but limiting the children included in the analysis to birth orders 2-4.For each of the above analyses, the country-level associations were meta-analyzed using the metan command in Stata. We also meta-analyzed the country-level associations by DHS region . We used random effects models for the meta-analysis to control for heterogeneity across countries. Stata Version 12 was used for the analyses.We present the results in two sections. First we show the neonatal and under-5 mortality odds ratios for short preceding birth intervals and then for long preceding birth intervals. We then run the same analyses stratified by mothers’ completed fertility. For both sets of analyses, we also present the results by region. The sample sizes and odds ratios by country are available in Additional File We found that children who are born after short birth intervals have increased odds of both neonatal and under-5 mortality compared to their siblings born after a regular birth interval. Having a <18 month interval had 82% increased odds (95% CI: 1.55-1.79) of neonatal mortality and 66% increased odds  of under-five mortality, compared to those born after 24-<60 month intervals. The <24 month interval had a slightly attenuated effect size of 61% increased odds  of neonatal mortality and 48% increased odds  of under-five mortality.  of neonatal mortality and 41% decreased odds (95% CI: 0.52-0.67) of under-five mortality compared to their siblings born after a 24-<60 month interval. The odds of mortality remained similar when examining birth interval ≥72 months.  and low completed fertility (four or fewer live births) mothers, even after controlling for confounders . The difAmong low completed fertility women, there were no statistically significant associations with neonatal mortality with short birth intervals. For under-five mortality, there was no statistically significant association for the <18 months  but a statistically significant association for the <24 months period . For the high completed fertility women, the odds ratios for neonatal and under-five mortality were statistically significant for both the <18 and <24 month intervals.  and neonatal mortality were not statistically significant in all regions. For under-5 mortality, there were mixed findings depending on the cut-off and region. In contrast, for high fertility women, the increased odds for neonatal and under-five mortality were statistically significant in all regions for both the <18 and <24 month intervals.  have higher odds of neonatal and under-five mortality, a finding that is consistent with most previous findings. However,While the overall effects of short intervals on neonatal and under-five mortality were consistent with findings from previous studies, our analyses also found major differences in odds ratios related to fertility history of mothers. When the associations were stratified by maternal completed fertility, the association between short birth intervals and child mortality largely disappeared for mothers with low completed fertility (had four or fewer live births at the end of their reproductive period). Furthermore, odds ratios tend to report larger magnitude associations than relative risks, a more easily interpretable measure of association. While weIn contrast, children of mothers with high completed fertility (had five or more live births by the end of their reproductive period) had strong associations between short intervals and mortality. However, when we limited the births included in the analysis for high fertility mothers to birth order 2-4, the magnitude of the associations were attenuated while remaining statistically significant. This suggests that there may be complex interactions between short birth intervals, differences across low- and high-fertility women, and parity.These findings seem to be consistent with the theory that maternal depletion is responsible for the higher mortality risks associated with short birth spacing.,7 Low feThe maternal depletion explanation is also consistent with our findings for longer birth intervals. The protective effect against mortality of long intervals was primarily seen among the high fertility women. However, among these women, the protective effective was similar comparing all births to just their earlier births; considering our previous theory regarding the cumulative adverse effect of short intervals, we expected the protective effect to also be smaller among the earlier births and greater for later births.Winkvist et al.  classifiIn contrast to our findings, previous studies have reported increased risk of neonatal and infant mortality associated with long birth intervals, even using cut-offs higher than the ones we used here. However, the previously reported adverse impact of long birth intervals may be largely affected by residual confounding. For instance, Rutstein  uses a cOur findings imply that lengthening birth intervals may only be physiologically beneficial to higher birth orders. Reproductive health programs may benefit more from targeting interventions to women with high parity. In contexts like India where there is a high prevalence of sterilization after a woman achieves her ideal of two or three children , lengtheOur study presents the impact of birth intervals on child mortality, using an innovative method of controlling for residual confounding by comparing short and long interval births to regular interval births of the same mother. Furthermore, other literature has not addressed or identified how maternal background characteristics modify the association between birth interval and mortality. This leads us to reevaluate previous findings on this topic, and reexamine the actual impact of short or long birth intervals on a child’s mortality risk.LiST, these analyses suggest that there is a causal linkage between short birth spacing and mortality in children. Other papers in this supplement will try to further identify this mechanism and help quantify the effects to be included in the model.For The authors declare that they have no competing interests.NK and NW both worked on all aspects of the work for this manuscript.Sample sizes and odds ratios for all outputs.Click here for file


WS070117 is a novel small molecule compound that significantly improves lipid metabolism disorders in high-fat-diet (HFD) induced hyperlipidemia in hamsters.14C] labeled acetic acid. Because WS070117 is an analog of adenosine, we evaluated the effect of WS070117 on AMP-activated protein kinase (AMPK) signaling. The results showed that the activation of AMPK in OLA-induced steatosis in HepG2 cells was up-regulated by treatment with 0.1, 1 and 10 μM WS070117. The hepatic cellular AMPK phosphorylation is also up regulated by WS070117 (6 and 18 mg/kg) treatment in HFD fed hamsters.We evaluated liver/body weight ratio, liver histology, serum and hepatic lipid content in HFD-fed hamsters treated with WS070117 for 8 weeks. Comparing with HFD fed hamsters, WS070117 (2 mg/kg per day and above) reduced serum triglyceride (TAG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C) and hepatic cholesterol and triglyceride contents. Oil Red O staining of liver tissue also showed that WS070117 improved lipid accumulation. We then carried out an experiment in the oleic acid (OLA)-induced steatosis model in HepG2 cell to investigate the lipid-lowering effect of WS070117. Oleic acid (0.25 mM) markedly induced lipid accumulation in HepG2 cells, but WS070117 (10 μM) inhibited cellular lipid accumulation. In OLA-treated HepG2 cells, WS070117 (above 1 μM) treatment reduced lipid contents which synthesized from [1-These new findings identify WS070117 as a novel molecule that regulates lipid metabolism in the hyperlipidemia hamster model. In vitro and in vivo studies suggested that WS070117 may regulate lipid metabolism through stimulating the activation of AMPK and its downstream pathways. Metabolism syndromes are often linked to the macronutrient content of the diet and there is evidence that excessive consumption of macronutrients such as carbohydrates, fats, and even protein may eventually lead to the development of metabolic syndromes . Diets hActivation of AMP-activated protein kinase (AMPK), a heterotrimeric energy-sensing protein, acts to restore cellular energy balance by promoting ATP-generating pathways such as fatty acid oxidation while simultaneously inhibiting ATP-utilizing pathways such as fatty acid synthesis . HistoriDuring our screening for AMPK activators, we serendipitously identified WS070117 as a lead compound with potent lipid-regulating properties. The objective of this study was to evaluate the impact of WS070117 on an animal and cellular model of hyperlipidemia. Studies in the hamster hyperlipidemia model revealed that WS070117 markedly improved lipid abnormalities associated with conditions characteristic of the high fat diet (HFD)-induced lipid metabolism disorder. Investigations into cellular signaling mechanisms suggested that WS070117 may produce the lipid-regulating effects by the phosphorylation of the AMPK signaling pathway.2',O3',O5'-tri-acetyl-N6-(3-hydroxylaniline)adenosine, named as WS070117 . Compound 1 was then dissolved in dry CH2Cl2:DMF (50:1) and a solution of SOCl2 in CH2Cl2 was added in a drop-wise manner. After dilution with CH2Cl2, the organic layer of the reaction mixture was washed with saturated NaHCO3 solution and brine and dried with anhydrous sodium sulfate. To a solution of O2',O 3',O5'-triacetyl-6-chloroadenosine in absolute ethanol was added 3-aminophenol and dry triethylamine. The mixture was then refluxed for 8 h at 60°C. The solution was concentrated in vacuum and the residue was chromatographed  to give WS070117 as a white solid.The synthesis of O7 Figure , began wad libitum.Golden hamsters (12-month-old) were purchased from Vital River Laboratory Animal Technology Co. Ltd. . All animal treatment procedures described in this study were approved by the Animal Care and Use Committee at the Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College . Animals were housed under well-controlled conditions of temperature (22 ± 2°C), humidity (55 ± 5%) and a 12 h light-dark cycle with access to food and water ad libitum for 8 weeks with the indicated drugs oral administration. WS070117 was dissolved in distilled water prior to administration. All drugs were given once daily by oral gavage for 8 weeks. The animals in the normal control and in the HFD model control group received the same volum of distilled water as that of the drug-treated groups.Except the control hamsters (n = 5), other hamsters (n = 40) were fed high fat diet (HFD) accustomedly for 2 weeks, blood was collected from orbit vein for assaying the content of serum total cholesterol and triglyceride for grouping. Then, The HFD fed hamsters were divided into the following groups with eight animals per group respectively, accounted into their body weight and the levels of serum total cholesterol and triglyceride, HFD model group, HFD+Simvastatin (2 mg/kg) as a positive group, HFD+WS070117 low dose group (2 mg/kg), HFD+WS070117 middle dose group (6 mg/kg) and HFD+WS070117 high dose group (18 mg/kg). The animals in normal control group were fed ordinary diet, and the other groups were still fed HFD 2 in air. After growth to 70% to 80% confluence, cells were rendered quiescent by incubating with serum-free medium for 12 h before fatty acid treatment. For OLA-induced lipid deposition, the medium for cells at 75% confluence was exchanged for new medium with or without 0.25 mmol/L OLA  ATP  radioactivity phosphorylated by a synthetic peptide substrate, SAMS  acetic acid  into lipid was performed as described previously [-14C] acetic acid (0.2 μCi/ml). Lipid was extracted with n-hexane:2-propanol  and dried under a nitrogen stream. Aliquots of lipid samples were loaded onto thin-layer chromatography plates and chromatographed in a solvent mixture containing hexane-diethylether-acetic acid . Bands corresponding to triglyceride and cholesteryl ester were scraped off, immersed in non-aqueous scintillation fluid, and counted for radioactivity.OLA HepG2 cells was co-incubated with 0.1,1 and 10 μM WS070117 for 12h. Incorporation of acetate in the presence of increasing concentrations of WS070117  induced lipid metabolism disorder in hamsters. Male Golden Syrian hamster, a widely used model in studying atherosclerosis and cholesterol metabolism in response to dietary fat manipulations, has many similar profiles in lipid metabolism compared to humans ,13. AfteThe nucleus of the parent molecule from which WS070117 is derived is an analogue of adenosine. Previously, several groups demonstrated that adenosine is an effective activator of AMPK ,16. In t14C]acetate in OLA-treated HepG2 cells. When compared with the well-known AMPK activator AICAR, WS070117 induced a similar profile of effects on lipid synthesis, with inhibition of cholesterol and TAG synthesis. We conclude that activation of AMPK induced by WS070117 could be responsible for its lipid lowering profile.To reinforce the lipid metabolism relevance of these cellular observations, we investigated the impact of WS070117 on OLA-treated HepG2 cells. Treatment of HepG2 cells with OLA induces morphological changes similar to those in steatotic hepatocytes ,22. HepaInhibition of hepatic lipogenesis is strongly associated with the anti-hepatic steatosis effect of WS070117. Endogenous OLA mediates the induction of de novo lipogenesis generally associated with intake of dietary saturated fat . ReducedThe present studies suggest that WS070117 may be used as a newer pharmacological agent to treat or control HFD-induced lipid metabolism disorder. Our results also support the view that the anti-hyperlipidemia and anti-steatosis effects of WS070117 may be attributed to the activation of AMPK and inhibition of de novo hepatic lipogenesis.The authors declare that they have no competing interests.ZL and HZ conceived the idea and designed the study. LH and SW were responsible for synthesis of WS070117. ZL and JG performed animal experiments, did the sampling, prepared the samples and carried out Western bolting analyses. KQ and JL participated in the experiments related to AMPK enzyme activity assay. ZL carried out the lipid metabolism assay experiments in animals and cultured cell, performed the statistical analysis and interpretation of the data. ZL drafted the manuscript and YL provided critical corrections to the manuscript. All authors read and approved the final manuscript.


To assess the reliability of maternally recalled birthweight and size in Entebbe, Uganda.The study population comprised 404 mothers, who were participants in the Entebbe Mother and Baby Study (EMaBS). Mothers were recruited to EMaBS during antenatal care, maternal characteristics were recorded during pregnancy, and birthweight was recorded at delivery. Four to seven years after delivery, mothers were asked to recall the child’s birthweight and size. Their responses were compared with the birthweight recorded in the EMaBS database.P = 0.04). Recalled and recorded birthweights showed moderate agreement with an intraclass correlation coefficient of 0.64. Four hundered mothers gave an estimate of birth size: the sensitivity and specificity of recalled birth size for classifying low birthweight were 76% (95% CI: 50–93%) and 70% (95% CI: 65–75%), respectively.Of 404 interviewed mothers, 303 (75%) were able to give an estimate of birthweight and for 265 of these EMaBS data on recorded birthweights were available. Women who were educated and whose children had low birth order were more likely to be able to give an estimate: 37 (14%) recalled the exact recorded birthweight; a further 52 (20%) were accurate to within 0.1 kg of the recorded weight. On average, mothers overestimated birthweight by 0.06 kg (95% CI: 0.00–0.13 kg, Mothers’ recall of birthweight was not precise but in absence of other data, recall of birthweight and size may have some value in epidemiological studies in these settings. Birthweight is an important predictor of future growth patterns . It is aIn developed countries, several studies have examined concordance between the birthweight recalled by the mother and the recorded birthweight and have shown that maternally recalled birthweight is a good proxy for recorded weight . HoweverBetween April 2003 and November 2005, the EMaBS birth cohort was established to investigate the effect of antihelminthic treatment during pregnancy on the offspring’s response to immunisation and on susceptibility to infectious diseases. Two thousand five hundred and seven women attending antenatal care at Entebbe hospital were enrolled into the trial. Full details of the trial design and procedures are described elsewhere . These data were linked with antenatal and delivery information from the EMaBS database, thus allowing for comparison of recalled and recorded birthweight.t-test. The intraclass correlation coefficient was calculated as a measure of the agreement between reported and recalled birthweight. Recorded birthweight was categorised into low birthweight (<2.5 kg), normal birthweight (2.5–4.0 kg) and large birthweight (>4.0 kg). Sensitivity and specificity of a mother’s perception of small birth size in detecting low birthweight babies and of a mother’s perception of large birth size in detecting large birthweight babies were calculated.Reliability of maternal recall of birthweight was assessed by calculating the mean difference between recalled birthweight and recorded birthweight and conducting a paired Logistic regression was used to examine factors associated with mother’s recall of birthweight. Two binary outcomes were considered: first, ability to recall any numerical estimate of birthweight; second, ability to recall birthweight to within 0.1 kg of the recorded weight. Explanatory factors considered were mother’s age, education and socio-economic status, child’s birth order, gender, recorded birthweight and the child’s age at the time of this study. Multivariable analysis was used to adjust for the possible confounding effect of factors that were crudely associated with the outcome.Between 21 September and 8 December 2010, 404 mothers were interviewed. Mothers who were interviewed were on average slightly older, were less likely to be primigravidae and had attended more routine study visits, than the remaining mothers enrolled in the EMaBS cohort whose children did not attend the clinic during this study period. Their children were less likely to have been born at home. Of the 404 children whose mothers were interviewed, 204 (51%) were male and 200 (49%) were female, with a mean age of 5.7 years . One hundred and ninety-seven (49%) had attended the clinic because of illness, and 207 (51%) had attended for a routine visit. The average age of mother at the time of delivery of the study baby was 25 years , and 356 (88%) said they still had the child health card. Three hundred and three (75%) of the women were able to give an estimate of birthweight; of the remaining 101 women who were unable to give an estimate of birthweight, 11 had delivered at home, and thus, birthweight is unlikely to have been measured . Characteristics of those who recalled and did not recall birthweight are shown in P = 0.04, paired t-test). Agreement between recalled and recorded birthweight was moderate  of the 404 interviewed mothers who gave birth in Entebbe hospital. Sixty-eight of these women were unable to recall their child’s birthweight, leaving 265 mother–child pairs with both a recalled and a recorded birthweight. The mean  of recalled and recorded birthweights were 3.28 kg  and 3.21 kg , respectively: on average, mothers overestimated the birthweight by 0.06 kg : the mean (SD) recorded birthweights for the recalled large, normal and small size groups were 3.73 kg (0.60 kg), 3.31 kg (0.40 kg) and 2.92 kg (0.49 kg), respectively. The sensitivity and specificity of mother’s recall of small size for low birthweight babies were 76% (95% CI: 50–93%) and 70% (95% CI: 65–75%), respectively, while the sensitivity and specificity for detecting large birthweight babies were 57% (29–82%) and 94% (91–97%), respectively.All but four of the 404 mothers gave a response regarding the size of the baby at birth. Thirty-five (9%) described their baby as large, 237 (59%) as normal and 128 (32%) as small. Reported size was associated with recorded birthweight  could not recall any numerical estimate of birthweight but almost all gave an approximate birth size. Those who gave an estimate of birthweight were not very accurate as to the precise figure, but there was moderate agreement between recalled and recorded birthweight. This was consistent with findings from the Netherlands by Studies by In this community, mothers’ concerns at birth are viability, absence of congenital anomalies and child’s sex and so little emphasis is placed on birthweight. Therefore, it is perhaps not surprising that many mothers in our study were unable to give a numerical estimate of weight. Ability to recall any numerical estimate of birthweight increased with education and decreased with the birth order of the child. However, we did not identify any maternal or child factors that were associated with accurate recall of birthweight. Similar results were reported by Mothers’ recall of birth size was more robust, allowing classification of babies as low birthweight with sensitivity and specificity of 76% and 70%, respectively (compared to a sensitivity of 60% and specificity of 93% seen in Cameroon .Possible sources of error and bias in this study were considered. Only one child per mother was enrolled into the EMaBS, and to be part of this analysis, the mother and child pair had attended the study clinic together; thus, it is unlikely that the mother gave data on any non-EMaBS sibling. Mothers who participated in this study were on average slightly older and had attended more routine study visits than members of the EMaBS cohort who were not included; and only 3% of their children had been born at home compared to EMaBS (In summary, mother’s recall of birthweight was not precise but in absence of other data, mother’s recalled birthweight and birth size have some value for epidemiological studies, as long as it is not crucial to know the exact birthweight. Clinicians and researchers using maternally recalled birthweight should be cautious when using such information. Recalled birth size should be used only where recorded birthweight is not available.


Background: Birth defects are a leading cause of neonatal mortality. Natural gas development (NGD) emits several potential teratogens, and U.S. production of natural gas is expanding.Objectives: We examined associations between maternal residential proximity to NGD and birth outcomes in a retrospective cohort study of 124,842 births between 1996 and 2009 in rural Colorado.Methods: We calculated inverse distance weighted natural gas well counts within a 10-mile radius of maternal residence to estimate maternal exposure to NGD. Logistic regression, adjusted for maternal and infant covariates, was used to estimate associations with exposure tertiles for congenital heart defects (CHDs), neural tube defects (NTDs), oral clefts, preterm birth, and term low birth weight. The association with term birth weight was investigated using multiple linear regression.Results: Prevalence of CHDs increased with exposure tertile, with an odds ratio (OR) of 1.3 for the highest tertile ; NTD prevalence was associated with the highest tertile of exposure , compared with the absence of any gas wells within a 10-mile radius. Exposure was negatively associated with preterm birth and positively associated with fetal growth, although the magnitude of association was small. No association was found between exposure and oral clefts.Conclusions: In this large cohort, we observed an association between density and proximity of natural gas wells within a 10-mile radius of maternal residence and prevalence of CHDs and possibly NTDs. Greater specificity in exposure estimates is needed to further explore these associations.http://dx.doi.org/10.1289/ehp.1306722Citation: McKenzie LM, Guo R, Witter RZ, Savitz DA, Newman LS, Adgate JL. 2014. Birth outcomes and maternal residential proximity to natural gas development in rural Colorado. Environ Health Perspect 122:412–417;  Some ofpelines)  or mutagpelines)  and are pelines) , raisingIn this analysis, we explored the association between maternal exposure to NGD and birth outcomes, using a data set with individual-level birth data and geocoded natural gas well locations. We conducted a retrospective cohort study to investigate the association between density and proximity of natural gas wells within a 10-mile radius of maternal residences in rural Colorado and three classes of birth defects, preterm birth, and fetal growth.Study population. We used information available in the publically accessible Colorado Oil and Gas Information System (COGIS) to build a geocoded data set with latitude, longitude, and year of development (1996–2009) for all gas wells in rural Colorado  oral cleft, including cleft lip with and without cleft palate as well as cleft palate  (b) NTD, including anencephalus, spina bifida without anecephaly, and encephalocele ; c) CHD, including transposition of great vessels, tetralogy of Fallot, ventricular septal defect, endocardial cushion defect, pulmonary valve atresia and stenosis, tricuspid valve atresia and stenosis, Ebstein’s anomaly, aortic valve stenosis, hypoplastic left heart syndrome, patent ductus arteriosis, coarctation of aorta, and pulmonary artery anomalies ; d) preterm birth (< 37 weeks completed gestation); e) term low birth weight ; and f) term birth weight as a continuous measure. Births with an oral cleft, NTD, or CHD were excluded from preterm birth and term low birth weight analysis. Preterm births were excluded from term birth weight analysis. Oral cleft, CHD, and NTD cases in the Colorado Responds to Children with Special Needs (CRCSN) birth registry, obtained from hospital records, the Newborn Genetics Screening Program, the Newborn Hearing Screening Program, laboratories, physicians, and genetic, developmental, and other specialty clinics (a) conotruncal defects ; b) endocardial cushion and mitrovalve defects ; c) pulmonary artery and valve defects ; d) tricuspid valve defects ; e) aortic artery and valve defects ; f) ventricular septal defects (VSD); and g) patent ductus arteriosis in births > 2,500 g  NTD, clinics  were mat 2,500 g .Exposure assessment. Distribution of the wells within a 10-mile radius of maternal residence shows 50% and 90% of wells to be within 2.3 and 7.7 miles of maternal residence, respectively. We used an inverse distance weighted (IDW) approach, commonly used to estimate individual air pollutant exposures from multiple fixed locations  for subsequent logistic and linear regression analysis. Each tertile was compared with the referent group .Statistical analysis. We used logistic regressions to study associations between each dichotomous outcome and IDW exposure group. We also considered term birth weight as a continuous outcome using multiple linear regression. First, we estimated the crude odds ratio (OR) associated with IDW exposure tertile for each binary outcome, followed by a Cochran–Armitage test to evaluate linear trends in binominal proportions with increasing IDW exposure . We further investigated associations by adjusting for potential confounders, as well as infant and maternal covariates selected based on both a priori knowledge and empirical consideration of their association with exposure and an outcome. Specifically, covariates in our analysis of all outcomes except outcomes with very few events  included maternal age, education , tobacco use , ethnicity , and alcohol use , as well as parity at time of pregnancy  and infant sex. Gestational age was also included in the analysis of term birth weight. Elevation of maternal residence also was considered in the analysis because most wells are < 7,000 feet, and elevation has been associated with both preterm birth and low birth weight .Births were approximately evenly divided between exposed and unexposed groups (0 wells in a 10-mile radius versus ≥ 1 well in a 10-mile radius) . EstimatBoth crude and adjusted estimates indicate a monotonic increase in the prevalence of CHDs with increasing exposure to NGD, as represented by IDW well counts . Births Prevalence of NTDs was positively associated with only the third exposure tertile, based on crude and estimated adjusted ORs for elevation . Births Both crude and adjusted estimates for preterm birth suggest a slight (< 10%) decreased risk of preterm birth with increasing exposure to NGD . Crude tWe observed a monotonic increase in the prevalence of NTDs with increasing exposure to NGD in our sensitivity analyses using 2- and 5-mile exposure radii as well as some attenuation in decreased risk for preterm birth and term low birth weight . Restricting births to 2000 through 2009, the period of most intense NGD in Colorado, attenuated the positive association between NTDs in the highest tertile and did not alter observed relationships for other birth outcomes .Exploratory analysis of CHDs by clinical diagnostic groups indicates increased prevalence of PAV defects by 60% , VSDs by 50% , and TVDs by 400%  in the most exposed tertile compared with those with no wells within a 10-mile radius .We found positive associations between density and proximity of natural gas wells within a 10-mile radius of maternal residence and birth prevalence of CHDs and possibly NTDs. Prevalence of CHDs increased monotonically from the lowest to highest exposure tertile, although even in the highest tertile the magnitude of the association was modest. Prevalence of NTDs was elevated only in the highest tertile of exposure. We also observed small negative associations between density and proximity of natural gas wells within a 10-mile radius of maternal residence and preterm birth and term low birth weight, and a small positive association with mean birth weight. We found no indication of an association between density and proximity of natural gas wells within a 10-mile radius of maternal residence and oral cleft prevalence.2, SO2, PM)  . NGD emiSO2, PM) . AmbientSO2, PM) . FurtherSO2, PM) .Two previous case–control studies have reported associations between maternal exposure to benzene and birth prevalence of NTDs and/or CHDs . The stu2, SO2, PM, and PAHs. A meta-analysis of four studies suggested associations of maternal NO2 and SO2 exposures with coarctation of the aorta and tetralogy of Fallot, and of maternal PM10 exposure with arterial septal defects  are not included. Birth weight and gestational age were obtained from birth certificates, which are generally accurate for birth weight and useful but less accurate for gestational age . Data onBecause of the rarity of specific birth defects in the study population, birth defects were aggregated into three general groups. This limited our study in that associations with specific birth defects may have been obscured. An exploratory analysis of CHDs by clinical diagnostic groups indicates increased prevalence of specific diagnostic groups  compared with aggregated CHDs .Another limitation of this study is the lack of temporal and spatial specificity of the exposure assessment. Because we did not have maternal residential history, we assumed that maternal address at time of delivery was the same as maternal address during the first trimester of pregnancy—the critical time period for formation of birth defects. Studies in Georgia and Texas estimate that 22–30% of mothers move residence during their pregnancy, and most mothers move within their locality , potentiSimilarly, we had consistent information only on existence of a well in the birth year. Lack of information on natural gas well activity levels, such as whether or not wells were producing or undergoing development, may have resulted in exposure misclassification. Actual exposure to natural gas–related pollutants likely varies by intensity of development activities. Lack of temporal and spatial specificity of the exposure assessment would most likely have tended to weaken associations . To addrThis study suggests a positive association between greater density and proximity of natural gas wells within a 10-mile radius of maternal residence and greater prevalence of CHDs and possibly NTDs, but not oral clefts, preterm birth, or reduced fetal growth. Further studies incorporating information on specific activities and production levels near homes over the course of pregnancy would improve exposure assessments and provide more refined effect estimates. Recent data indicate that exposure to NGD activities is increasingly common. The COGCC estimates that 26% of the > 47,000 oil and gas wells in Colorado are located within 150–1,000 feet of a home or other type of building intended for human occupancy . Taken t(143 KB) PDFClick here for additional data file.


Ciliated hepatic foregut cyst (CHFC) is a rare lesion which originates from detached hepatic diverticulum or from detachment and migration of buds from the esophageal and bronchial regions of the foregut which subsequently get entrapped by the liver during the early embryonic development of the foregut. CHFCs are mostly seen in adults and are rarely reported in children, with only about 10 cases reported in this age group. Hereby, we present two cases of CHFC in two 3.5-year-old boys; one of them had cystic lesion at medial segment of left lobe of liver (common site), and in the other one it was located at right lobe of liver (less common site). Histologically, both cysts had four layers composed of inner ciliated, pseudostratified, columnar epithelium; subepithelial connective tissue; smooth muscle layer; and an outer fibrous layer. Ciliated hepatic foregut cyst (CHFC), a rare foregut developmental malformation, usually presents as a solitary, unilocular, or occasionally multilocular cyst in the liver –9. This A 3.5-year-old boy presented with right upper quadrant abdominal pain. At the ninth month of gestation, a liver cyst was detected by US (no photographs were available). No further investigation was done after birth. Physical examination was normal. Laboratory tests showed mild microcytic anemia ; the other tests, including liver function tests, were within normal limits. Ultrasonography and CT showed a simple 3.7 × 2.8 cm subcapsular liver cyst in median segment of left lobe with no enhancement at CT . He undeA 3.5-year-old boy was referred to the hospital with constipation. Physical examination was normal. A hepatic cyst was detected by US. Hematologic test showed mild normocytic anemia . Serum electrolytes and liver function test were within normal limits. Ultrasonography showed a heterogeneous lesion, measuring 3.6 × 3.2 cm in right lobe of liver , and CT At histological examination, both cysts had four typical layers of ciliated hepatic foregut cyst, composed of inner ciliated, pseudostratified, columnar epithelium; subepithelial connective tissue; smooth muscle layer; and an outer fibrous layer Figures . Foci ofThe incidence of hepatic cysts is approximately 5% in general population; they are classified into parasitic and nonparasitic cysts. One of the uncommon types of liver cyst is CHFC, which arises from the embryonic foregut. It is believed that it originates from a detached hepatic diverticulum or from detached and migrated foregut buds, entrapped in liver at the early embryonic development of the foregut . BecauseThese lesions are extremely rare in children and only 10 cases have been reported  1, 4–6,,6, 12–15The mean age at diagnosis is 50 years (3 months to 82 years), with a slight male predominance , 10, 19.The clinical course is usually benign; however, malignant transformation to squamous cell carcinoma with aggressive behavior has been reported in 3% of cases, all of which happened in adults between 21 and 51 years of age, supporting the idea of excision of the cyst when discovered in adults, and careful clinical followup of the patients , 16–18. Since hydatid cyst is endemic in our country, it is usually at the top of the differential diagnoses list for a liver cyst, such as case 2. Inclusion of CHFC in the differential diagnosis and performance of serological tests to exclude hydatid cyst could make laparoscopic removal more likely with less postoperative complications. We have presented two cases of CHFC in 3.5-year-old boys due to extreme rarity of this lesion in children. CHFC in children is not previously reported from Iran.


Objectives. Newfoundland and Labrador (NL) has one of the highest incidences of Type 1 diabetes mellitus (T1DM) worldwide. Rates of T1DM are increasing and the search for environmental factors that may be contributing to this increase is continuing. Methods. This was a population-based case control design involving the linkage of data from a diabetes database with live birth registration data. 266 children aged 0–15 years with T1DM were compared to age- and gender-matched controls. Chi-square analysis and multivariate conditional logistic regression were carried out to assess maternal and infant factors . Results. Cases of T1DM were more likely to be large-for-gestational-age (P = 0.024) and delivered by C-section (P = 0.009) as compared to controls. C-section delivery was associated with increased risk of T1DM  when birth weight and gestational age were included in the model, but not when size-for-gestational-age was included . Conclusions. Birth by C-section was found to be a risk factor for the development of T1DM in a region with high rates of T1DM and birth by C-section. These findings may have an impact on health practice, health care planning, and future research. T1DM is thought to originate through a combination of genetic and unknown environmental factors, of which environmental factors remain poorly defined. Newfoundland and Labrador (NL), Canada, is recognized as having one of the highest rates of T1DM worldwide [Type 1 diabetes mellitus (T1DM) is one of the most common chronic diseases in childhood and results from autoimmune destruction of pancreatic orldwide . T1DM isThis study was a case control design involving the linkage of data extracted from the Newfoundland and Labrador Diabetes Database (NLDD) with the Live Birth System (LBS). The NLDD is maintained by the Janeway Pediatric Research Unit at the Janeway Child Health Care Centre (JCHCC) in St. John's, NL. The JCHCC is the only tertiary care pediatric hospital in the province. The NLDD includes data for the majority of cases of T1DM diagnosed in NL from 1987 to present. Children are included in the database as part of a prospective provincial study on the epidemiology of T1DM in NL. They have a confirmed diagnosis of T1DM according to Canadian Diabetes Association (CDA) criteria . The LBSPatient records were individually linked across datasets. Cases included children born in NL from 1992 onwards which have been diagnosed with T1DM before 15 years of age. Children with type 2 diabetes, maturity-onset diabetes of youth, transient hyperglycemia, and diabetes caused by chemotherapy or cystic fibrosis are excluded from the NLDD and thus were not included in the study. Children born prior to 1992 were not included in the study because there are no electronic birth notification data available before this date. N = 798) were selected for each case matched on year of birth, sex, and health authority of mother's residence at time of delivery. Power analysis was performed to determine whether this sample size would be sufficient to detect statistical significant associations between T1DM and the risk factors of interest. The power analysis was conducted considering an overall rate of birth by C-section in NL as 30.9% [A unique identifier, such as the provincial health insurance plan number, was not available for all children in the NLDD. As a result case subjects were linked to the LBS using child's date of birth and mother's maiden name. Where available, child's name was used to verify the linkage. Of the 301 cases in the NLDD, 23 were excluded because they were born out of province. Of the remaining 278, 6 were excluded as duplicate records. Linkage was possible for all but six children resulting in a total of 266 cases included in the study. Three control subjects  or not . Cases and controls were also classified as small/appropriate-for-gestational-age or large-for-gestational-age using the method described by Kramer et al. . Method Descriptive statistics were generated to describe the distribution of cases and controls. Demographic and clinical factors of mothers, including age, marital status, education, place of residence, parity (number of live born children delivered), and complications of pregnancy, were included. Cases and controls were analyzed by sex, place of residence, age of onset, length of gestation, type of delivery, birth weight, size for gestational age, and birth order. Chi-square tests were used to predict diabetes status on the basis of the independent variables. Conditional multiple logistic regression was used to assess the relationship between T1DM risk and the variables of interest. Two conditional logistic regression models were employed. The first model contained birth weight, gestational age, parity, delivery method, mother's marital status, mother's education level, mother's age, maternal hypertension, and mother's T1DM status. The second model incorporated all variables in the first model with the exception of birth weight and gestational age which were replaced with size-for-gestational-age. Birth weight and gestational age were not included in the same model as size-for-gestational-age as they are components of this variable. The Statistical Package for the Social Sciences (SPSS) 17.0 was used to generate descriptive statistics and chi-squares. SAS 9.2 was used to conduct the conditional multivariate logistic regressions.This study received approvals from the Human Investigation Committee of Memorial University, from each of the hospital boards and the Secondary Uses Committee of the Newfoundland and Labrador Centre for Health Information (NLCHI) prior to commencement.P = 0.024). It was more common for cases to be delivered by C-section than controls . T1DM was more common among first or second born cases compared to those born third or higher (P = 0.022).P = 0.015). In the second model, which included size-for-gestational-age, C-section delivery was not associated with increased risk of T1DM . Both parity and size-for-gestational-age were found to be significant risk factors for T1DM from chi-square analysis  . TheorieP = 0.03) [P < 0.001). For other maternal factors, such as education level and marital status, there were no associations found which is consistent with other similar studies [In the present study, maternal age at time of birth was not found to be significantly associated with risk of T1DM in offspring; other studies have found significant relationships between mother's age and T1DM risk. A recent meta-analysis of 37 studies found that the odds of T1DM increased by 10% for children whose mothers were over 35 years of age at time of birth  . Convers = 0.03)  found th studies , 15. The studies , 16, 17; studies  did not There was no significant relationship between parity and T1DM risk found in the current study. This is different than a Western Australia population based cohort study of 835 cases of T1DM diagnosed by the age of 15 that found a significant decrease in T1DM with increasing birth order . P < 0.05) [Birth weight and gestational age were not found to be associated with risk of T1DM in the present study; however, chi-squared analysis revealed a significant difference between T1DM and size-for-gestational-age with a higher percentage of cases than controls born large-for-gestational-age, but this was no longer significant in the conditional logistic regression models. Our findings do not support the findings of a meta-analysis of 11 studies examining birth weight which found that a birth weight greater than 4,000 grams was associated with an increased odds of T1DM  . Finding < 0.05) . However < 0.05)  found th < 0.05) .The findings of this study should be considered in the context of its strengths and weaknesses. An important strength is that the use of a record linkage case-control study design eliminates recall bias that is apparent in cross-sectional study designs. Secondly, data contained in the LBS were collected at time of birth by healthcare professionals, and the NLDD data were collected from physician charts at time of T1DM diagnosis which contributes to the reliability of the data. A limitation of this study is that there was very little information available pertaining to fathers as the majority of the information collected at the time of birth for the LBS is related to the mother and child. Thus, paternal factors and family history could not be considered for analysis. This study identified C-section as a significant risk factor for the development of T1DM among children aged 0–15 years in NL, a region with very high rates of T1DM. Findings may have an impact on health practice, health care planning and future research related to T1DM among children. Further research should be undertaken to understand the nature of this association.


Gastric duplication cysts are rare variety of gastrointestinal duplications. Sometimes they may present with complications like hemorrhage, infection, perforation, volvulus, intussusception and rarely neoplastic changes in the gastric duplication cyst. We present one and half year old male child who developed sudden abdominal distension with pain and fever for two days. Ultrasound revealed a cystic mass in the hypochondrium and epigastric regions. On exploration an infected and perforated gastric duplication cyst was found. Surgical excision of most part of cyst wall with mucosal stripping of the rest was performed. Histopathology confirmed the diagnosis of gastric duplication cyst. Early surgical intervention can result in good outcome. Gastric duplication cysts constitute about 2-7 % of all gastrointestinal duplications. Majority of gastric duplication cysts are large and non-communicating. Most of the cases of gastric duplication cysts present in early age, however, in some cases patient may remain asymptomatic for a long period and might present with sudden onset of abdominal distension, pain, signs of obstruction, peritonitis etc. . In thisA male baby of one and half year presented in emergency with fever, abdominal pain and distension for two days. Fever was high grade and not associated with rigors and chills. Pain abdomen was continuous and patient had with few bouts of non bilious, non projectile vomiting. The past medical and surgical history was unremarkable. General physical examination revealed temp 101o F, pulse 100/min and respiratory rate 30/min. On abdominal examination generalized rigidity and guarding were present. No mass or viscera could be palpated. Bowel sounds were audible. A clinical diagnosis of acute peritonitis was made. X-ray abdomen was unremarkable and ultrasound showed a cystic structure of 4cm x 5cm size in the left hypochondrium and epigastrium. Urgent laparotomy was planned after initial stabilization.At operation a cyst having intimate contact with the greater curvature of the stomach was found Fig. . It was Gastric duplications are very uncommon congenital anomalies. They usually arise at greater curvature of stomach. Almost all gastric duplications are cystic in nature. Usually they are non-communicating. In our case gastric duplication was at greater curvature, cystic in nature and non-communicating. The clinical presentations of gastric duplication cysts depends upon site, size, communication with part of the alimentary tract and associated complications  , 3.In complicated cases patient may present with an acute abdomen, peritonitis or even pancreatitis. Other complications include hemorrhage, infection, perforation of the cyst and compression on surrounding structures. Clinical features thus vary. Our patient presented with fever, pain and abdominal distension suggestive of complications like infection and perforation of the cyst  , 5.The differential diagnoses of gastric duplication cyst include omental cyst, mesenteric cyst, choledochal cyst, ovarian cyst, hydronephrosis etc. Ultrasound abdomen, contrast GI studies, CT-scan and MRI are helpful diagnostic modalities. In our case ultrasound abdomen did show a cyst  , 5.The treatment of gastric duplication cyst is surgical resection of the cyst. Due to its close connection with the adjacent gut usually it is very difficult to completely excise therefore partial resection with the mucosal stripping of the remaining cyst is recommended as done in present case. The diagnosis of duplication cyst is based upon presence of GIT mucosa in cyst with smooth muscle coat in the wall. Cyst must have an intimate contact with any part of GIT. In our patient all these features were present. Early diagnosis and prompt surgical intervention with optimal surgical procedure carries a good prognosis  , 3 , 6.Source of Support: NilConflict of Interest: None declared


The preterm birth rate varies greatly between different migrant populations in high income countries, but the reasons have not been well elucidated . War refTo investigate the association between duration of residence and preterm birth risk among war refugees, we exploited data from the Swedish national registers. We included information on all live singletons births between 1992 and 2008 of women during their first 5 years of residency in Sweden, who were at least 13 years of age when they had obtained Swedish residency and had immigrated from Yugoslavia (1992–2000), Iraq (1992–2004), Somalia (1992–2004), or Afghanistan (1992–2004) with a spouse of the same origin. Information on perinatal indicators and maternal age was retrieved from the Swedish Medical Birth Register (SMBR) and country of birth and year of immigration from the Register of the Total Population. Those with a birth weight for gestational age >3 standard deviations (SDs) or <−6 SDs according to the Marsal growth chart [Number of years since immigration were categorized into 0, 1, or 2–4 years. Information on maternal age, parity, mode of delivery, diagnoses during pregnancy and delivery, birth weight, gestational age, and sex of offspring were retrieved from SMBR. Diagnoses were coded according to the Swedish version of the International Classification of Diseases. The ninth revision (ICD-9) was used between 1987 and 1996, and the tenth revision (ICD-10) has been used thereafter. Gestational age was estimated by early second trimester ultrasound with reference to the Marsal growth chart . If ultrTo estimate the association between duration of residence and preterm birth, a logistic regression model was used, adjusting for maternal country of origin, age, parity and sex of the infant. We further used multinomial logistic regression to estimate ORs for very preterm birth and moderately preterm birth, adjusting for the abovementioned covariates. All statistical analyses were performed using STATA version 12 for Windows. The study was approved by the regional ethics committee in Stockholm.Compared with the second year of residency, the OR of preterm birth (<37 weeks) was 1.39 (95 % CI 1.13–1.72) in the first year of residency and 1.10 (95 % CI 0.94–1.28) in the third to fifth year of residency. Compared with the second year of residency, the ORs of very and moderately preterm birth in the first year of residence were 2.15 (95 % CI 1.37–3.38) and 1.23 (95 % CI 0.98–1.57), respectively  axis. High levels of corticotrophin-releasing hormone (CRH) are very common, while cortisol levels vary much more, depending on the duration of stress . High plWe recognize that stress is not the only possible explanation for the higher risk of preterm birth in our study population. One may hypothesize that other exposures in the country of origin, such as malnutrition and infectious agents may also be involved, as well as informal barriers to care in the resettlement country.We also found an increased risk of very preterm but not moderately preterm birth in the third to fifth year of residency. This confirms a time pattern after migration that has previously been demonstrated in immigrants in Canada in a longer time scale , althougOur study is limited by the lack of precise information on the timing of migration in our study population; we lacked information on when the refugees set foot on Swedish soil, and the exact date of when residency was acquired. Thus, our estimates probably underestimate the time dependency of preterm birth in relation to migration. The sample size also restricted further investigation of potential mechanisms of preterm birth.In conclusion, this study demonstrates an increased risk of preterm birth in war refugees during the first year of residency in Sweden. This suggests that different levels of stress, including post-traumatic stress, might explain some of the differences in rates of preterm birth between migrant populations. Further studies with more specific measures of stress and hormonal profiles are needed to investigate stress as a risk factor for preterm birth in migrant populations.


Src-associated in mitosis  has been implicated in the oncogenesis and progression of several human cancers. The aim of this study was to investigate the clinicopathologic significance of Sam68 expression and its subcellular localization in colorectal cancer (CRC).Sam68 expression was examined in CRC cell lines, nine matched CRC tissues and adjacent noncancerous tissues using reverse transcription (RT)-PCR, quantitative RT-PCR and Western blotting. Sam68 protein expression and localization were determined in 224 paraffin-embedded archived CRC samples using immunohistochemistry. Statistical analyses were applied to evaluate the clinicopathologic significance.P = 0.033), advanced T stage (P < 0.001), N stage (P = 0.023) and distant metastasis (P = 0.033). Sam68 nuclear localization correlated significantly with poor differentiation (P = 0.002) and T stage (P =0.021). Patients with high Sam68 expression or Sam68 nuclear localization had poorer overall survival than patients with low Sam68 expression or Sam68 cytoplasmic localization. Patients with high Sam68 expression had a higher risk of recurrence than those with low Sam68 expression.Sam68 was upregulated in CRC cell lines and CRC, as compared with normal tissues; high Sam68 expression was detected in 120/224 (53.6%) of the CRC tissues. High Sam68 expression correlated significantly with poor differentiation (Overexpression of Sam68 correlated highly with cancer progression and poor differentiation in CRC. High Sam68 expression and Sam68 nuclear localization were associated with poorer overall survival. Colorectal cancer (CRC) is one of the most prevalent malignancies worldwide. Although advances have been made in diagnostic and therapeutic techniques, the prognosis of CRC patients with distant metastases still remains poor . Thus, cSrc-associated in mitosis, 68 kDa (Sam68) belongs to the signal transduction and activation of RNA (STAR) family of K homology (KH) domain-containing RNA binding proteins  and is oThe above mentioned biological functions of Sam68 closely linked this protein to oncogenic properties. First, Sam68 is involved in promotion of cell cycle progression, cell proliferation, transformation, tumorigenesis and metastasis in different cellular context -25. Seco2 at 37°C.Colorectal cancer cell lines including LS174t, Colo205, SW480, HT29, HCT116 and SW620 cells were cultured in RPMI 1640  supplemented with 10% fetal bovine serum, 100 μg/μL streptomycin and 100 μg/μL penicillin in a humidified incubator containing 5% COFor the use of clinical materials for research purposes, prior patients' consents and approval were obtained from the Sun Yat-sen University and Cancer Center Institutional Board. All samples were collected and analyzed with prior written informed consents from the patients. A total of 224 paraffin-embedded colorectal cancer samples, which were histopathologically and clinically diagnosed at the Sun Yat-sen University Cancer Center between the year 2000 to 2003, were examined. All of the patients had received chemotherapy after surgery. Prior patient consent and approval were obtained from the Institutional Research Ethics Committee.The clinicopathological features of the patients are summarized in Additional file -∆∆Ct method as described by Livak et al. [Total RNA was extracted from the cultured cells and surgical tissues using TRIzol reagent (Invitrogen) according to the manufacturer's instructions. Reverse transcription polymerase chain reaction (RT-PCR) and quantitative real-time polymerase chain reaction (Q-PCR) analysis of Sam68 expression were performed as previously described , using pk et al.  and normTissue and cell lysates were prepared using SDS lysis buffer and the protein concentration was determined using the Bradford assay . Equal amounts of protein were separated by electrophoresis on a 10.5% sodium dodecyl sulfate polyacrylamide gel and electrotransferred from the gel to a nitrocellulose membrane. After blocking with 5% milk solution in Tris-buffered saline with Tween (TBST) for 1 hour, the membrane was incubated with primary antibody against rabbit antibody Sam68  and rabbit anti-α-Tubulin  primary antibodies. After washing with TBS-T, the membrane was incubated with secondary antibody against rabbit immunoglobulin G or mouse immunoglobulin G; then, it was examined with the enhanced chemiluminescence detection system  according to the manufacturer’s instructions.Paraffin sections were deparaffinized with xylene and rehydrated, then submerged into EDTA antigenic retrieval buffer and microwaved for antigenic retrieval. The sections were then treated with 3% hydrogen peroxide in methanol to quench the endogenous peroxidase activity, followed by incubation with 1% BSA to block the non-specific binding. The sections were incubated with rabbit anti-Sam68  overnight at 4°C. As negative controls, rabbit anti-Sam68 antibody was replaced with normal goat serum, or the rabbit anti-Sam68 antibody was blocked by co-incubation with a recombinant Sam68 polypeptide at 4°C overnight prior to the immunohistochemical staining. The staining intensity was scored on a scale of 0 to 3 as 0 (no staining), 1 (weak staining ~ light yellow), 2 (moderate staining ~ yellowish brown) or 3 (strong staining ~ brown). Tumors with a staining intensity ≥ 2 in which at least 50% of the malignant cells were Sam68-positive were classified as high expression; tumors with a staining intensity < 2 or in which less than 50% of the malignant cell were Sam68-positive were classified as low expression.t-test. The association between Sam68 expression and clinicopathological variables were assessed using the Mann–Whitney U test. Survival curves were plotted using the Kaplan-Meier method. The Cox proportional hazards regression model was used for univariate and multivariate analysis. Two-tailed P values < 0.05 were considered significant.All statistical analyses were carried out using SPSS 10.0 . The significance of the differences between the normal and tumor tissue Q-PCR results were assessed using the Student’s two-tailed We examined the expression of Sam68 using Western blotting in seven human colon cancer cell lines and two cases of normal intestine tissues. The results displayed that Sam68 protein expression level was much higher in CRC cell lines than that in normal intestine tissues Figure A. We nexP < 0.01, Student’s t-test).Western blotting and RT-PCR analyses were performed to determine the expression of Sam68 in nine paired primary CRC tissues and the matched adjacent non-cancerous tissues. Sam68 was significantly upregulated at both the protein Figure A and mRNIn agreement with the Western blotting results, immunohistochemical analysis confirmed that Sam68 was overexpressed in all nine of the CRC tissues tested, compared with the paired adjacent normal tissues Figure D. Taken We further performed immunohistochemical analysis to determine the expression patterns of Sam68 in 224 paraffin-embedded CRC tissues and 43 lymph node metastatic tissues. Negative to moderate Sam68 staining was detected in the adjacent normal tissues , advanced T stage (P < 0.001), N stage (P = 0.023), and distant metastasis (P = 0.033) in this cohort of 224 cases of CRC. In addition, high expression level of Sam68 was significantly associated with the nuclear localization of Sam68 (P = 0.012). Moreover, Nuclear localization of Sam68 correlated significantly with tumor differentiation (P = 0.002) and advanced T stage (P = 0.021). These observations suggest that increased expression of Sam68 or nuclear localization of Sam68 was closely associated with aggressive phenotypes of CRC.Statistic analyses were performed to evaluate the expression patterns of Sam68 and the clinicopathological features of CRC. As shown in Table P < 0.001). Kaplan-Meier survival analysis demonstrated that patients with low levels of Sam68 expression had significantly longer median survival than patients with low Sam68 expression . The 5-year survival rate for patients with low Sam68 expression was 70% , compared to 54%  for patients with high Sam68 expression.In univariate Cox regression analysis, tumor differentiation, T stage, N stage and distant metastasis were significant prognostic factors in this cohort of CRC patients , as the median overall survival time of patients expressing low levels of Sam68 in the metastatic lymph nodes was significantly longer than patients expressing high levels of Sam68 in the metastatic lymph nodes .Next, we analyzed the relationship between the expression of Sam68 in metastatic lymph nodes and survival. Kaplan-Meier analysis indicated that the expression level of Sam68 in the metastatic lymph nodes had a significant impact on survival . Additionally, Kaplan-Meier survival analysis confirmed that the median survival time for patients with Sam68 nuclear localization was significantly shorter than patients with Sam68 cytoplasmic localization . The 5-year survival rate for patients with Sam68 cytoplasmic localization was 70% , compared to 54%  for patients with Sam68 nuclear localization. The Sam68 expression level, subcellular localization of Sam68, pathological stage and N stage were identified as independent prognostic factors for overall survival in CRC in multivariate survival analysis  or with well/moderately differentiated tumors, both the Sam68 expression level .Click here for fileUnivariate Cox regression analysis of potential prognostic factors for CRC patients.Click here for fileMultivariate Cox regression analysis of Sam68 levels and other potential prognostic factors for CRC patients.Click here for fileMultivariate Cox regression analysis of Sam68 localization and other potential prognostic factors for CRC patients.Click here for file


Objective. Gestational diabetes mellitus (GDM) influences fetal development and offspring's metabolic risk. We evaluated this association in 17-year-old offspring adjusting for birth weight (BW) and prepregnancy maternal BMI (mBMI). Study Design. The JPS birth cohort contains extensive data on 92,408 births from 1964 to 1976. Offspring's BMI and blood pressure (BP) were obtained from military records. For a subcohort born between 1974 and 1976, prepregnancy mBMI was available. Offspring were classified as born to mothers with GDM (n = 293) or born to mothers without recorded GDM . Results. GDM offspring had higher mean BMI and systolic and diastolic BP compared to no-recorded-GDM offspring. After adjusting for BW, GDM remained significantly associated with offspring BMI and diastolic BP . In the subcohort, when prepregnancy mBMI was entered to the models, it markedly attenuated the associations with GDM. Conclusions. Maternal characteristics have long-term effects on cardiometabolic outcomes of their offspring aged 17 years. Fetal development is regulated by maternal and fetal characteristics that influence the intrauterine environment. The adaptation of the fetus to intrauterine conditions results in fetal “programming” which may also determine the metabolic “fate” of the fetus later in life . Gestational diabetes mellitus (GDM) is defined as carbohydrate intolerance with different degrees of severity, first diagnosed during pregnancy . It is tIntrauterine hyperglycaemic conditions influence fetal metabolism and in turn may influence later life morbidity. The short-term consequences of GDM for the offspring are high birth weight  and assoThere is some evidence that neonatal birth weight , maternaIn a previous report  on a subIn this study, we aimed to assess the long-term implications of maternal GDM on adolescent BMI and blood pressure, among JPS offspring at age 17, after taking into account different characteristics, such as offspring birth weight and prepregnancy mBMI.The Jerusalem perinatal study (JPS) is a population-based cohort that includes all births occurred in western Jerusalem and its surroundings between 1964 and 1976. Detailed information on data collection has been previously described . In brieData on maternal conditions, obstetric complications, and interventions during labor and delivery, including GDM status, were collected systematically in 92,408 deliveries. The original data collection instruments included rubrics for GDM  and pre-gestational diabetes . In this study, we used only the definition for GDM health condition and did not included the pregestational diabetes cases. In that era, all pregnant women were screened for glycosuria at each antenatal visit, and if found positive, they would be referred for an oral glucose tolerance test.Postpartum interviews of a subset of mothers  who gave birth between November 1974 and the end of 1976 provided further information on medical conditions and lifestyle habits, including prepregnancy mBMI, weight gain during pregnancy, gestational age at delivery, and smoking habits. These interviews were conducted at the bedside on the first or second day postpartum by nurse-midwives and captured 98% of the births.Through the Israeli Population Registry, we verified the identities of 99% of the offspring and 96.2% of the mothers in the JPS cohort using their unique identity (ID) numbers. The ID number and other identifiers enabled us to link the JPS data with military draft records. This linkage provided information regarding the weight, height, and systolic and diastolic blood pressure which were measured in 60,191 JPS singleton offspring  at age 17.The following variables were included in the analysis: parents' ages at birth, birth order, offspring's birth weight (continuous variables), maternal level of education  and socioeconomic status (SES) using a scale based on husband occupation . Maternal ethnic origin was classified according to her father's country of birth . Maternal medical history included data on diabetes and pre-eclampsia in the current pregnancy or in other pregnancies . Information on maternal smoking during pregnancy, gestational age at delivery , prepregnancy mBMI, and weight gain during pregnancy was available only in the subcohort of the mothers who gave birth between 1974 and 1976. 2). The examiners were blinded to the perinatal data.Anthropometric and blood pressure information measured in the offspring at age 17 were obtained from the Israel Defense Forces medical database. Details about these examinations have been described elsewhere . BrieflyThe current study examined the differences in offspring BMI and blood pressure measured at age 17 among the two groups: offspring whose mothers had GDM in the index pregnancy (GDM) and offspring whose mothers did not have GDM in any of her pregnancies during the period 1964–1976 and recorded in the JPS (no-recorded-GDM). We included only singleton pregnancies in this analysis.Baseline characteristics of the study population and variables obtained at age 17 are presented for the two groups of offspring categorized by their mother's GDM status. Descriptive analysis was used to compare the BMI and blood pressure mean values for the offspring groups, and linear regression analysis was used to adjust for other characteristics in multivariate models. For the 1964–1976 subcohort, two sets of models were examined: (1) model 1 included the maternal GDM status as the main covariate (GDM compared to the no-recorded-GDM group) and (2) model 2 additionally adjusted for offspring birth weight. These models were also adjusted for the following covariates: maternal age at birth, ethnic origin (Israel as a reference group), level of education (0–8 years as a reference group), maternal pre-eclampsia (in one or more pregnancies between 1964 and 1976), birth order, SES (middle class as a reference group), and offspring gender.For the subcohort of 1974–1976, we further adjusted for prepregnancy mBMI and weight gain during the pregnancy (model 3). In the 1974–1976 analysis, maternal smoking and gestational age at delivery were also included in the models as covariates. To account for the correlation between siblings within families, the multivariate analysis for the entire cohort (1964–1976) was repeated using a mixed linear model (SAS PROC MIXED). Both approaches provided similar findings; and therefore, the results from the linear regression models are presented. Analyses were conducted with SPSS version 14.0  and SAS package version 9.1 .Of the 92,408 deliveries recorded in the JPS between 1964 and 1976, military induction examination information was available for 60,191 (65.1%) singleton offspring. Of these, 293 (0.5%) were born to mothers with GDM and 59,499 (98.8%) to mothers with No-recorded-GDM. An additional 399 (0.7%) offspring were born to mothers who had GDM not in the index pregnancy but in one of her other pregnancies during the period 1964–1976 and recorded in the JPS. These offspring were not included in the analysis. Of the 16,912 singleton deliveries during the 1974–1976 subcohort, military information was available for 11,412 offspring, of whom 77 offspring were born to GDM mothers, and 11,335 to no-recorded-GDM mothers. P < 0.001). Mothers in the GDM group were more educated and from higher SES status compared to mothers from the no-recorded-GDM group. In addition, mothers with GDM were more likely to have pre-eclampsia in one of their pregnancies recorded in the JPS, compared to mothers belonging to the no-recorded-GDM group.P < 0.05 for all outcomes).BMI, systolic and diastolic blood pressure mean values were significantly higher in GDM offspring as compared to the mean values obtained in offspring born to no-recorded-GDM mothers  independent of BW (model 2). In the analysis of the 1974–1976 subcohort, when models were further adjusted for maternal smoking and gestational age at delivery, GDM was positively and significantly associated with offspring BMI measured at age 17 (model 1); however, when birth weight was included in the model (model 2), it attenuated the association with GDM. When prepregnancy mBMI and weight gain during pregnancy were included in the model (model 3), the association of GDM with the offspring BMI was attenuated markedly. When we evaluated the association of GDM with systolic blood pressure measured in offspring at age 17, no significant associations were found in the entire cohort and in the subcohort as well (model 1). When prepregnancy mBMI included in the analysis of the subcohort, it was positively associated with systolic blood pressure (model 3). In distinction to systolic blood pressure, GDM was significantly associated with diastolic blood pressure in the entire cohort, even after introducing birth weight to the model (model 2). In the subcohort models, there was no evidence of an association between GDM and offspring diastolic blood pressure, and prepregnancy mBMI was positively associated with diastolic blood pressure at age 17.Our study implies that maternal characteristics pre- and during pregnancy and the intrauterine environment are important factors for future cardiometabolic conditions of their offspring. In the entire JPS cohort of 1964–1976, we found that the contribution of maternal GDM to offspring BMI in young adulthood is independent of neonatal birth weight. These findings are consistent with previous studies –18. HoweOur study suggested a positive association between maternal GDM and diastolic blood pressure measured in the 17-years-old offspring, in the whole cohort. Other studies, focusing on childhood blood pressure, have demonstrated higher blood pressure values in offspring of mothers with GDM , 9, whilEven though the association between GDM and diastolic blood pressure did not reach statistical significance in the smaller subcohort (1974–1976), the coefficient of GDM was attenuated when prepregnancy mBMI was included in the model. This implies that the association between maternal GDM and offspring diastolic blood pressure in young adulthood might as well be accounted for prepregnancy mBMI. There is growing evidence in the literature, that prepregnancy mBMI is a strong predictor of offspring health status later in life , 19–21. This study has potential limitations that should be considered. During the JPS data collection period (1964–1976), screening for GDM was not routine in Israel and the prevalence of GDM in the cohort was lower (0.5%) than the current reported prevalence of 3%–5%, possibly due to differences both in the diagnosis of GDM, and in the study population, as 85% of the mothers had a pre pregnancy BMI less than 25 at that time. In addition, we do not have information regarding GDM severity or mode of treatments of the mothers. We assume that the more severe cases of GDM were ascertained and diagnosed at that time; therefore, our results may represent the associations for the offspring of a group of mothers with a more severe form of GDM rather than the GDM detected today by screening during pregnancy. Another limitation is that military induction examination information was available for only 65.1% of the offspring included in the JPS cohort. In the Israeli Defense Force, military service is compulsory for all Jewish males and females, but female subjects are less commonly recruited to army service due to religious belief and practice, so the proportion of females with available information is lower than that of males (38% versus 62%). In addition, citizens may be exempted if they are religiously observant or have physical or mental disabilities. Therefore, the ultra-orthodox and disabled population may be underrepresented in this study. We believe that this underrepresentation has potential effect on the point estimates, but due to the composition of our study population, it does not alter the inspected associations.The findings from the JPS cohort add to the knowledge regarding the long term effects of maternal prepregnancy characteristics on cardiometabolic outcomes in offspring aged 17 years, and point to populations at risk. Given the fact that in the US, 13.9% to 25.1% of births are to obese mothers [