Alicia Granada

Human exposure to endocrine-disrupting chemicals and prenatal risk factors for cryptorchidism and hypospadias: a nested case-control study

Background Exposure to xenoestrogens during pregnancy may disturb the development and function of male sexual organs. Objective In this study we aimed to determine whether the combined effect of environmental estrogens measured as total effective xenoestrogen burden (TEXB) is a risk factor for male urogenital malformations. Methods In a case–control study, nested in a mother–child cohort (n = 702) established at Granada University Hospital, we compared 50 newborns with diagnosis of cryptorchidism and/or hypospadias with 114 boys without malformations matched by gestational age, date of birth, and parity. Controls did not differ from the total cohort in confounding variables. TEXB and levels of 16 organochlorine pesticides were measured in placenta tissues. Characteristics of parents, pregnancy, and birth were gathered by questionnaire. We used conditional and unconditional regression models to estimate odds ratios (ORs) and 95% confidence intervals (CIs). Results TEXB from organohalogenated compounds was detectable in 72% and 54% of case and control placentas, respectively. Compared with controls, cases had an OR for detectable versus non-detectable TEXB of 2.82 (95% CI, 1.10–7.24). More pesticides were detected in cases than in controls (9.34 ± 3.19 vs. 6.97 ± 3.93). ORs for cases with detectable levels of pesticides, after adjusting for potential confounders in the conditional regression analysis, were o,p′-DDT (OR = 2.25; 95% CI, 1.03–4.89), p,p′-DDT (OR = 2.63; 95% CI, 1.21–5.72), lindane (OR = 3.38; 95% CI, 1.36–8.38), mirex (OR = 2.85; 95% CI, 1.22–6.66), and endosulfan alpha (OR = 2.19; 95% CI, 0.99–4.82). Engagement of mothers in agriculture (OR = 3.47; 95% CI, 1.33–9.03), fathers’ occupational exposure to xenoestrogens (OR = 2.98; 95% CI, 1.11–8.01), and history of previous stillbirths (OR = 4.20; 95% CI, 1.11–16.66) were also associated with risk of malformations. Conclusions We found an increased risk for male urogenital malformations related to the combined effect of environmental estrogens in placenta.

Oestrogenicity of paper and cardboard extracts used as food containers

Bisphenol-A (BPA), dibutyl phthalate (DBP), and di-2-ethylhexyl phthalate (DEHP), which are common chemical residues in food-packaging materials, were investigated in paper and cardboard containers used for take-away food. The oestrogenicity of aqueous extracts was tested in E-Screen bioassay and analysis carried out by high-performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GC/MS). Oestrogenicity was demonstrated in 90% of extracts (geometric mean [GM] = 11.97 pM oestradiol equivalents g−1). DEHP, DBP, and BPA (GM = 341.74, 37.59, and 2.38 ng g−1 of material) were present in 77.50, 67.50, and 47.50% of samples, respectively. In bivariate analyses, no significant association was found between the levels of these chemicals and oestrogenicity in cardboard/paper extracts. A close-to-significant association was found between oestrogenicity and DBP (β = 1.25; p = 0.06) in paper extracts, which reached statistical significance in multivariate analysis (β = 1.61; p = 0.03). Paper and cardboard used in food packaging may contribute to the inadvertent exposure of consumers to endocrine-disrupting chemicals.

Organochlorine pesticides in umbilical cord blood serum of women from Southern Spain and adherence to the Mediterranean diet

Exposure of pregnant women to organochlorine (OC) pesticides largely derives from contaminated food, but environmental, occupational, and domestic factors have also been implicated. We investigated the presence of nine OC residues in the umbilical cord blood of newborns in Southern Spain and analyzed the relationship of this exposure with maternal and pregnancy variables, including maternal adherence to the Mediterranean Diet (MD). OCs were detected in 95% of umbilical cord blood samples from the 318 mothers, who had a mean degree of adherence to the MD of 56.77 (SD: 16.35) (range, 0-100). The MD prioritizes consumption of vegetable and fruit over meat and dairy products, and OCs are generally lipophilic molecules that accumulate in foods of animal origin. Consumption of meat, fish, and dairy products was associated with dichlorodiphenyldichloroethylene (DDE) in umbilical cord serum, and dairy product intake with lindane. Vegetable consumption was also associated with lindane and fruit intake with endosulfan I. We found no significant association between MD adherence and the presence of OC residues in serum. However, closer adherence to the MD may offer greater protection against OC exposure because of its reduced content in meat and dairy products.

Dietary exposure assessment of pregnant women to bisphenol-A from cans and microwave containers in Southern Spain

Bisphenol-A (BPA), material for polycarbonate and epoxy resin synthesis, has been detected in canned food, among other food containers. In mammal studies, BPA transferred from mother to fetus, caused abnormality of reproductive organs, and advanced female puberty. BPA from canned food and microwave containers was analyzed using high performance liquid chromatography and gas chromatography-mass spectrometry. Population was cohort of mother-son pairs established at Granada University Hospital. Frequency of food consumption (including canned food) was studied with a semi-quantitative questionnaire. The most frequently consumed products were fish and juice cans, consumed 1-3 times/week by 34.7% and 22.3% of the study population, respectively. The women made little use of polymer microwave containers, 52.8% never using them and 45.9% using them <3 times/month. Estimated mean (standard deviation) intake of BPA was 1.1(0.839) microgram/day. No relationship was found between BPA exposure and maternal socio-demographic variables or newborn characteristics. This study offers the first estimate of BPA dietary intake by pregnant women in Southern Spain. The consumption of canned foods and drinks by these women means that their exposure was lower than EFSA estimates for the European population. Nevertheless it remains of concern, given the proven undesirable effects of low-level exposure and higher susceptibility of pregnant women.

Proposal of a Mediterranean diet index for pregnant women

Numerous studies have addressed the nutritional needs of pregnant women. The nutritional status of the woman before and during gestation affects the growth of the fetus and the course of the pregnancy and influences the risk of obesity for mother and infant. The aim of this study was to propose a diet quality index for pregnancy based on a Mediterranean-type diet (MDS-P), evaluating the diet of a group of pregnant women by applying the Mediterranean Diet Score (MDS) and evaluating their intake of micronutrients required in optimal amounts during pregnancy, such as Fe, folic acid and Ca. The data used to construct this index (MDS-P) were gathered by means of a FFQ specifically designed for pregnant women. The mean MDS of this group, was 4.31 (sd 1.32), considered to represent satisfactory compliance with the Mediterranean diet (range 0-8). The mean MDS-P (range 0-11), which also takes account of dietary intake or supplements of folic acid, Fe and Ca was 7.53 (sd 1.44), indicating a compliance of around 70 %. The present study findings suggest that the MDS-P, which evaluates the adequacy of folic acid, Fe and Ca as well as compliance with the Mediterranean diet, may represent a valid tool for the specific assessment of the diet of pregnant women living in countries in the Mediterranean area. Further studies are required to complete the validation process.

Assessment of the total effective xenoestrogen burden in extracts of human placentas.

We have standardized a method to assess the total effective xenoestrogen burden (TEXB) in human placentas by the extraction and separation by high-performance liquid chromatography of two fractions containing lipophilic xenoestrogens (alpha) and endogenous hormones (beta), followed by assessing their estrogenicity in MCF-7 breast cancer cell-based E-Screen and Yeast Estrogen Screen (YES) bioassays. The means of TEXB alpha concentrations (in estradiol equivalent (Eeq) units) were 1.32 and 0.77 Eeq pM g−1 placenta in the E-Screen and YES, respectively; TEXB beta concentrations were 6.97 and 11.56 Eeq pM g−1 placenta, respectively. The interclass correlation coefficient was low and a fair level of agreement was observed after kappa test correction. According to the E-Screen and YES, TEXB alpha was ≥LOD in 70.0 and 55.0% of the placentas and 92.5 and 82.5% in beta, respectively. Although both bioassays can be recommended for assessing TEXB, there is greater experience with the use of the E-Screen for estrogenic assessment after extensive extraction of complex human matrices.

Exposición humana a disruptores endocrinos

Con el fin de contribuir a la definicion de un plan de manejo para la Reserva Provincial Acambuco, ubicada en la Selva Tucumano Boliviana (Noroeste Argentino, S 22o 10’ 3,6” W 63o 54’ 43,2“), por medio de actividades escolares y entrevistas a cazadores y otros habitantes de la reserva (n= 92), se ubicaron y se mapearon los sitios de alimentacion del tigre (Panthera onca), los lugares donde esta especie fue cazada y se verifico si el area protegida esta en condiciones de proporcionarle refugio y alimentacion suficientes, teniendo en cuenta el impacto ambiental de las principales actividades economicas de sus habitantes. El proceso de roza y quema para instalar cultivos, la explotacion forestal sin ningun orden y principalmente el sobrepastoreo, explican que los 22 sitios de caza de subsistencia actuales, coincidentes con los lugares de alimentacion del tigre y que los 7 sitios donde esta especie fue capturada entre 1976 y 2001, esten todos afuera de la reserva. Esto permite concluir en que el mal estado de conservacion del area protegida no permite la existencia del tigre dentro de ella, por lo que debe implementarse un plan de manejo que abarque toda su area de influencia.