Assessing Dietary Pesticide Intake and Potential Health Effects: The Application of Global Metabolomics Analysis.

Scientific information is not yet available to provide insight into how individual metabolome might be affected by the presence of pesticides in regular diets. This study aimed to evaluate the perturbation of metabolomic pathways in children who switched their diets from conventional foods to mostly organic foods for five consecutive days. We selected 46 child-matched spot urine samples with distinct differences of urinary pesticide metabolite levels between the conventional and organic eating days and then analyzed those urine samples on three analytical platforms to perform global metabolomics analysis. We found statistically significant perturbations of metabolic pathways relevant to inflammation, oxidative stress, and the demands of xenobiotic detoxification when children switched their conventional diets to mostly organic foods. The outcomes of this study allow us to extend the current understanding beyond organophosphate pesticides' acute toxicity of cholinesterase inhibition to the perturbation of metabolic pathways at dietary intake levels.

Dietary risk of neonicotinoid insecticides through fruit and vegetable consumption in school-age children.

Although the systemic property of neonicotinoid (neonics) has become the most widely used insecticide worldwide since late 1990s, the current literature offers limited information about the human dietary intake and the potential risks of neonics. In this study, we aimed to assess the cumulative risk of total neonics intakes through fruit and vegetable consumption in 58 children ages 8-12 participated in the Hangzhou China (HZC) study over 5 consecutive weekends. Individual neonic residues in each food item were aggregated using the relative potency factor approach into a single metric (IMIRPF), representing dietary intakes of imidacloprid-equivalent total neonics. We then estimated the average daily intake (ADI) of total neonics through fruit and vegetable consumption and evaluated the cumulative dietary risk of neonics. All of the 123 samples were detected with at least one neonic. Commonly consumed foods, such as carrots, green vegetables, baby cabbage, and apple were found with more than 6 neonics. The estimated ADIs of total neonics vegetable and fruit consumption using the mean IMIRPF for apples and green vegetables, two most consumed food items, were 237.1*10-6 and 106.8*10-6 mg/kg/day, respectively. Although the estimated ADIs were below the current chronic reference dose (cRfD) of imidacloprid, we have stipulated the possible future downward revision of cRfD. The potential health risk of neonics to children via dietary exposure should raise more public concern considering the increase use of neonics and the ubiquitous presence in fruits and vegetables.

Characterization of Daily Dietary Intake and the Health Risk of Neonicotinoid Insecticides for the U.S. Population.

Although neonicotinoids have been the most commonly used insecticides globally, very limited data related to their dietary intake and health risks are available. In this study, we used the relative potency factor approach to aggregate individual neonicotinoids into a single metric (IMIRPF) representing the intakes of total neonicotinoids in relation to imidacloprid for each food item. We then estimated the average daily intake (ADI) of neonicotinoids using residue data collected from U.S. Congressional Cafeteria study (USCC) and USDA/PDP and food consumption data from NHANES 2011-2012. Among the USCC and USDA/PDP samples, squash (427.2 ng/g) and spinach (569.2 ng/g), had the highest average IMIRPF, respectively. The estimated ADIs were below the current chronic reference dose (cRfD) for imidacloprid. However, due to their wide use, it is logical to expect the ubiquity of neonicotinoids in foods. Therefore, the importance of conducting routine dietary intake assessment for neonicotinoids should not be ignored.

Neonicotinoid Residues in Fruits and Vegetables: An Integrated Dietary Exposure Assessment Approach.

Neonicotinoids have become the most widely used insecticides in the world since introduced in the mid 1990s, yet the extent of human exposure and health impacts is not fully understood. In this study, the residues were analyzed of seven neonicotinoids in fruit and vegetable samples collected from two cross-sectional studies: the U.S. Congressional Cafeteria study (USCC) and the Hangzhou China (HZC) study. We then employed a relative potency factor method to integrate all neonicotinoids in each food sample using the respective reference dose values as the basis for summation. The findings were compared with data published by the U.S. Department of Agriculture Pesticide Data Program (USDA/PDP). Imidacloprid and thiamethoxam were the most commonly detected neonicotinoids in fruits and vegetables with 66 and 51% detection in the HZC study and 52 and 53% detection in the USCC study, respectively. The overall frequency of detection for neonicotinoids in the USDA/PDP samples was much lower than those reported here for the USCC or HZC studies, with imidacloprid being the most frequently detected neonicotinoid at 7.3%. The high frequencies of neonicotinoid detection in fruits and vegetables in the USCC and HZC studies give us a snapshot of the ubiquity of neonicotinoid use in global agriculture and make it clear that neonicotinoids have become part of the dietary staple, with possible health implications for individuals.

PTEN is required to maintain luminal epithelial homeostasis and integrity in the adult mammary gland.

In the mammary gland, PTEN loss in luminal and basal epithelial cells results in differentiation defects and enhanced proliferation, leading to the formation of tumors with basal epithelial characteristics. In breast cancer, PTEN loss is associated with a hormone receptor-negative, basal-like subtype that is thought to originate in a luminal epithelial cell. Here, we show that luminal-specific PTEN loss results in distinct effects on epithelial homeostasis and mammary tumor formation. Luminal PTEN loss increased proliferation of hormone receptor-negative cells, thereby decreasing the percentage of hormone receptor-positive cells. Moreover, luminal PTEN loss led to misoriented cell divisions and mislocalization of cells to the intraluminal space of mammary ducts. Despite their elevated levels of activated AKT, Pten-null intraluminal cells showed increased levels of apoptosis. One year after Pten deletion, the ducts had cleared and no palpable mammary tumors were detected. These data establish PTEN as a critical regulator of luminal epithelial homeostasis and integrity in the adult mammary gland, and further show that luminal PTEN loss alone is not sufficient to promote the progression of mammary tumorigenesis.

Residential Exposure to Pesticide During Childhood and Childhood Cancers: A Meta-Analysis.

CONTEXT: There is an increasing concern about chronic low-level pesticide exposure during childhood and its influence on childhood cancers. OBJECTIVE: In this meta-analysis, we aimed to examine associations between residential childhood pesticide exposures and childhood cancers. DATA SOURCES: We searched all observational studies published in PubMed before February 2014 and reviewed reference sections of articles derived from searches. STUDY SELECTION: The literature search yielded 277 studies that met inclusion criteria. DATA EXTRACTION: Sixteen studies were included in the meta-analysis. We calculated effect sizes and 95% confidence intervals (CIs) by using a random effect model with inverse variance weights. RESULTS: We found that childhood exposure to indoor but not outdoor residential insecticides was associated with a significant increase in risk of childhood leukemia (odds ratio [OR] = 1.47; 95% CI, 1.26-1.72; I(2) = 30%) and childhood lymphomas (OR = 1.43; 95% CI, 1.15-1.78; I(2) = 0%). A significant increase in risk of leukemia was also associated with herbicide exposure (OR = 1.26; 95% CI, 1.10-1.44; I(2) = 0%). Also observed was a positive but not statistically significant association between childhood home pesticide or herbicide exposure and childhood brain tumors. LIMITATIONS: The small number of studies included in the analysis represents a major limitation of the current analysis. CONCLUSIONS: Results from this meta-analysis indicated that children exposed to indoor insecticides would have a higher risk of childhood hematopoietic cancers. Additional research is needed to confirm the association between residential indoor pesticide exposures and childhood cancers. Meanwhile, preventive measures should be considered to reduce children's exposure to pesticides at home.

Mammary Stem Cells and Tumor-Initiating Cells Are More Resistant to Apoptosis and Exhibit Increased DNA Repair Activity in Response to DNA Damage.

Adult stem cells and tumor-initiating cells (TICs) often employ different mechanisms of DNA damage response (DDR) as compared to other tissue cell types. However, little is known about how mammary stem cells (MaSCs) and mammary TICs respond to DNA damage. Using the mouse mammary gland and syngeneic p53-null tumors as models, we investigated the molecular and physiological consequences of DNA damage in wild-type MaSCs, p53-null MaSCs, and p53-null TICs. We showed that wild-type MaSCs and basal cells are more resistant to apoptosis and exhibit increased non-homologous end joining (NHEJ) activity. Loss of p53 in mammary epithelium affected both cell-cycle regulation and DNA repair efficiency. In p53-null tumors, we showed that TICs are more resistant to ionizing radiation (IR) due to decreased apoptosis, elevated NHEJ activity, and more-rapid DNA repair. These results have important implications for understanding DDR mechanisms involved in both tumorigenesis and therapy resistance.

Intratumoral heterogeneity in a Trp53-null mouse model of human breast cancer.

UNLABELLED: Intratumoral heterogeneity correlates with clinical outcome and reflects the cellular complexity and dynamics within a tumor. Such heterogeneity is thought to contribute to radio- and chemoresistance because many treatments may target only certain tumor cell subpopulations. A better understanding of the functional interactions between various subpopulations of cells, therefore, may help in the development of effective cancer treatments. We identified a unique subpopulation of tumor cells expressing mesenchymal-like markers in a Trp53-null mouse model of basal-like breast cancer using fluorescence-activated cell sorting and microarray analysis. Both in vitro and in vivo experiments revealed the existence of cross-talk between these "mesenchymal-like" cells and tumor-initiating cells. Knockdown of genes encoding ligands upregulated in the mesenchymal cells and their corresponding receptors in the tumor-initiating cells resulted in reduced tumorigenicity and increased tumor latency. These studies illustrate the non-cell-autonomous properties and importance of cooperativity between tumor subpopulations. SIGNIFICANCE: Intratumoral heterogeneity has been considered one important factor in assessing a patient's initial response to treatment and selecting drug regimens to effectively increase tumor response rate. Elucidating the functional interactions between various subpopulations of tumor cells will help provide important new insights in understanding treatment response and tumor progression.

Identification of a novel nonsense mutation in the rod domain of GFAP that is associated with Alexander disease.

Alexander disease (AxD) is an astrogliopathy that primarily affects the white matter of the central nervous system (CNS). AxD is caused by mutations in a gene encoding GFAP (glial fibrillary acidic protein). The GFAP mutations in AxD have been reported to act in a gain-of-function manner partly because the identified mutations generate practically full-length GFAP. We found a novel nonsense mutation (c.1000 G>T, p.(Glu312Ter); also termed p.(E312*)) within a rod domain of GFAP in a 67-year-old Korean man with a history of memory impairment and leukoencephalopathy. This mutation, GFAP p.(E312*), removes part of the 2B rod domain and the whole tail domain from the GFAP. We characterized GFAP p.(E312*) using western blotting, in vitro assembly and sedimentation assay, and transient transfection of human adrenal cortex carcinoma SW13 (Vim(+)) cells with plasmids encoding GFAP p.(E312*). The GFAP p.(E312*) protein, either alone or in combination with wild-type GFAP, elicited self-aggregation. In addition, the assembled GFAP p.(E312*) aggregated into paracrystal-like structures, and GFAP p.(E312*) elicited more GFAP aggregation than wild-type GFAP in the human adrenal cortex carcinoma SW13 (Vim(+)) cells. Our findings are the first report, to the best of our knowledge, on this novel nonsense mutation of GFAP that is associated with AxD and paracrystal formation.

The intraflagellar transport protein ift80 is essential for photoreceptor survival in a zebrafish model of jeune asphyxiating thoracic dystrophy.

PURPOSE. Jeune's asphyxiating thoracic dystrophy (JATD) is an autosomal recessive disorder with symptoms of retinal degeneration, kidney cysts, and chondrodysplasia and results from mutations in the ift80 gene. This study was conducted to characterize zebrafish lacking ift80 function for photoreceptor degeneration and defects in ciliogenesis to establish zebrafish as a vertebrate model for visual dysfunction in JATD and to determine whether ift80 interacts genetically with Bardet-Biedl syndrome (BBS) genes. METHODS. Zebrafish were injected with morpholinos (MOs) targeted to the ift80 gene. Retinas were analyzed by histology, transmission electron microscopy, and immunohistochemistry. Ear and kidney cilia were analyzed by whole-mount immunostaining. Intraflagellar transport (IFT) particle composition was subjected to Western blot analysis. Genetic interactions were tested by coinjection of MOs against ift80 and bbs4 or bbs8 followed by in situ hybridization. RESULTS. Zebrafish lacking ift80 function exhibited defects in photoreceptor outer segment formation and photoreceptor death. Staining with opsin antibodies revealed opsin mislocalization in both rods and cones. Ultrastructural analysis showed abnormal disc stacking and shortened photoreceptor outer segments. The kinocilia of the ear and motile cilia in the kidney were shorter and reduced in number. Western blot analysis revealed a slight increase in the stability of other IFT proteins. Coinjection of MOs against ift80 and BBS genes led to convergent-extension defects. CONCLUSIONS. Zebrafish lacking ift80 exhibited defects characteristic of JATD. Because the developing outer segments degenerated, Ift80 could possibly act as a maintenance factor for the IFT particle.

Recombinant porcine lactoferrin expressed in the milk of transgenic mice protects neonatal mice from a lethal challenge with enterovirus type 71.

The human Enterovirus genus of the piconavirus family causes most of the febrile illnesses that affect children during the summer season in Taiwan. Enterovirus type 71 (EV71) plays a key role in patients with hand-foot-and-mouth disease (HFMD) combined with severe paralysis or encephalitis. It is important to find a method for preventing infection with EV71 since there is no antiviral agent or vaccine for humans. In this study, we developed a transgenic mouse model for demonstrating the protective effects of recombinant lactoferrin (LF) against EV71 infection. Transgenic mice carrying alpha-lactalbumin-porcine lactoferrin (alphaLA-pLF) and BALB/c wild-type mice were subjected to EV71 inoculation. First, we analyzed the expression efficiencies of recombinant pLF (rpLF) in hemizygous and homozygous transgenic mice. Following EV71 inoculation on the 4th day of life, pups ingesting transgenic milk showed the significantly higher survival rate and heavier body weight compared to wild-type mice. RT-PCR analysis for EV71 viral RNA showed that the recombinant pLF had a blocking effect on EV71 infection. Our data suggest that oral intake of pLF-enriched milk exhibited the ability to prevent infection with EV71. The study also provides an animal model for validating the protective effects of pLF.